@article{muthusamy_williams_o'toole_brudvig_adler_weimer_muddiman_ghashghaei_2022, title={Phosphorylation-dependent proteome of Marcks in ependyma during aging and behavioral homeostasis in the mouse forebrain}, volume={1}, ISSN={["2509-2723"]}, url={http://dx.doi.org/10.1007/s11357-022-00517-3}, DOI={10.1007/s11357-022-00517-3}, abstractNote={Ependymal cells (ECs) line the ventricular surfaces of the mammalian central nervous system (CNS) and their development is indispensable to structural integrity and functions of the CNS. We previously reported that EC-specific genetic deletion of the myristoylated alanine-rich protein kinase C substrate (Marcks) disrupts barrier functions and elevates oxidative stress and lipid droplet accumulation in ECs causing precocious cellular aging. However, little is known regarding the mechanisms that mediate these changes in ECs. To gain insight into Marcks-mediated mechanisms, we performed mass spectrometric analyses on Marcks-associated proteins in young and aged ECs in the mouse forebrain using an integrated approach. Network analysis on annotated proteins revealed that the identified Marcks-associated complexes are in part involved in protein transport mechanisms in young ECs. In fact, we found perturbed intracellular vesicular trafficking in cultured ECs with selective deletion of Marcks (Marcks-cKO mice), or upon pharmacological alteration to phosphorylation status of Marcks. In comparison, Marcks-associated protein complexes in aged ECs appear to be involved in regulation of lipid metabolism and responses to oxidative stress. Confirming this, we found elevated signatures of inflammation in the cerebral cortices and the hippocampi of young Marcks-cKO mice. Interestingly, behavioral testing using a water maze task indicated that spatial learning and memory is diminished in young Marcks-cKO mice similar to aged wildtype mice. Taken together, our study provides first line of evidence for potential mechanisms that may mediate differential Marcks functions in young and old ECs, and their effect on forebrain homeostasis during aging.}, number={4}, journal={GEROSCIENCE}, publisher={Springer Science and Business Media LLC}, author={Muthusamy, Nagendran and Williams, Taufika I and O'Toole, Ryan and Brudvig, Jon J. and Adler, Kenneth B. and Weimer, Jill M. and Muddiman, David C. and Ghashghaei, H. Troy}, year={2022}, month={Jan} }
 @article{dinh_paudel_brochu_popowski_gracieux_cores_huang_hensley_harrell_vandergriff_et al._2020, title={Inhalation of lung spheroid cell secretome and exosomes promotes lung repair in pulmonary fibrosis}, volume={11}, ISSN={["2041-1723"]}, url={http://dx.doi.org/10.1038/s41467-020-14344-7}, DOI={10.1038/s41467-020-14344-7}, abstractNote={Abstract}, number={1}, journal={NATURE COMMUNICATIONS}, publisher={Springer Science and Business Media LLC}, author={Dinh, Phuong-Uyen C. and Paudel, Dipti and Brochu, Hayden and Popowski, Kristen D. and Gracieux, M. Cyndell and Cores, Jhon and Huang, Ke and Hensley, M. Taylor and Harrell, Erin and Vandergriff, Adam C. and et al.}, year={2020}, month={Feb} }
 @misc{sheats_yin_fang_park_crews_parikh_dickson_adler_2019, title={MARCKS and Lung Disease}, volume={60}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2018-0285TR}, abstractNote={Abstract MARCKS (myristoylated alanine‐rich C kinase substrate) is a prominent PKC substrate expressed in all eukaryotic cells. It is known to bind to and cross‐link actin filaments, to serve as a bridge between Ca2+/calmodulin and PKC signaling, and to sequester the signaling molecule phosphatidylinositol 4,5‐bisphosphate in the plasma membrane. Since the mid‐1980s, this evolutionarily conserved and ubiquitously expressed protein has been associated with regulating cellular events that require dynamic actin reorganization, including cellular adhesion, migration, and exocytosis. More recently, translational studies have implicated MARCKS in the pathophysiology of a number of airway diseases, including chronic obstructive pulmonary disease, asthma, lung cancer, and acute lung injury/acute respiratory distress syndrome. This article summarizes the structure and cellular function of MARCKS (also including MARCKS family proteins and MARCKSL1 [MARCKS‐like protein 1]). Evidence for MARCKS's role in several lung diseases is discussed, as are the technological innovations that took MARCKS‐targeting strategies from theoretical to therapeutic. Descriptions and updates derived from ongoing clinical trials that are investigating inhalation of a MARCKS‐targeting peptide as therapy for patients with chronic bronchitis, lung cancer, and ARDS are provided.}, number={1}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Sheats, Mary K. and Yin, Qi and Fang, Shijing and Park, Joungjoa and Crews, Anne L. and Parikh, Indu and Dickson, Brian and Adler, Kenneth B.}, year={2019}, month={Jan}, pages={16–27} }
 @article{adler_2018, title={Does transforming growth factor-beta induce persistent airway obstruction after asthma exacerbations?}, volume={58}, DOI={10.1165/rcmb.2017-0339ed}, number={5}, journal={American Journal of Respiratory Cell and Molecular Biology}, author={Adler, K. B.}, year={2018}, pages={543–544} }
 @article{brudvig_cain_sears_schmidt-grimminger_wittchen_adler_ghashghaei_weimer_2018, title={MARCKS regulates neuritogenesis and interacts with a CDC42 signaling network}, volume={8}, ISSN={["2045-2322"]}, DOI={10.1038/s41598-018-31578-0}, abstractNote={Abstract}, journal={SCIENTIFIC REPORTS}, author={Brudvig, J. J. and Cain, J. T. and Sears, R. M. and Schmidt-Grimminger, G. G. and Wittchen, E. S. and Adler, K. B. and Ghashghaei, H. T. and Weimer, J. M.}, year={2018}, month={Sep} }
 @article{dinh_cores_hensley_vandergriff_tang_allen_caranasos_adler_lobo_cheng_2017, title={Derivation of therapeutic lung spheroid cells from minimally invasive transbronchial pulmonary biopsies}, volume={18}, ISSN={1465-993X}, url={http://dx.doi.org/10.1186/s12931-017-0611-0}, DOI={10.1186/s12931-017-0611-0}, abstractNote={Resident stem and progenitor cells have been identified in the lung over the last decade, but isolation and culture of these cells remains a challenge. Thus, although these lung stem and progenitor cells provide an ideal source for stem-cell based therapy, mesenchymal stem cells (MSCs) remain the most popular cell therapy product for the treatment of lung diseases. Surgical lung biopsies can be the tissue source but such procedures carry a high risk of mortality.In this study we demonstrate that therapeutic lung cells, termed "lung spheroid cells" (LSCs) can be generated from minimally invasive transbronchial lung biopsies using a three-dimensional culture technique. The cells were then characterized by flow cytometry and immunohistochemistry. Angiogenic potential was tested by in-vitro HUVEC tube formation assay. In-vivo bio- distribution of LSCs was examined in athymic nude mice after intravenous delivery.From one lung biopsy, we are able to derive >50 million LSC cells at Passage 2. These cells were characterized by flow cytometry and immunohistochemistry and were shown to represent a mixture of lung stem cells and supporting cells. When introduced systemically into nude mice, LSCs were retained primarily in the lungs for up to 21 days.Here, for the first time, we demonstrated that direct culture and expansion of human lung progenitor cells from pulmonary tissues, acquired through a minimally invasive biopsy, is possible and straightforward with a three-dimensional culture technique. These cells could be utilized in long-term expansion of lung progenitor cells and as part of the development of cell-based therapies for the treatment of lung diseases such as chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF).}, number={1}, journal={Respiratory Research}, publisher={Springer Nature}, author={Dinh, Phuong-Uyen C. and Cores, Jhon and Hensley, M. Taylor and Vandergriff, Adam C. and Tang, Junnan and Allen, Tyler A. and Caranasos, Thomas G. and Adler, Kenneth B. and Lobo, Leonard J. and Cheng, Ke}, year={2017}, month={Jun} }
 @article{yin_fang_park_crews_parikh_adler_2016, title={An Inhaled Inhibitor of Myristoylated Alanine-Rich C Kinase Substrate Reverses LPS-Induced Acute Lung Injury in Mice}, volume={55}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2016-0236rc}, abstractNote={Intratracheal instillation of bacterial LPS is a well-established model of acute lung injury (ALI) and/or acute respiratory distress syndrome (ARDS). Because the myristoylated alanine-rich C kinase substrate (MARCKS) protein is involved in neutrophil migration and proinflammatory cytokine production, we examined whether an aerosolized peptide that inhibits MARCKS function could attenuate LPS-induced lung injury in mice. The peptide, BIO-11006, was delivered at 50 μM via inhalation either just before intratracheal instillation of 5 μg of LPS into Balb/C mice, or 4, 12, 24, or 36 hours after LPS instillation. Effects of BIO-11006 were evaluated via analysis of mouse disease-related behavior, lung histology, bronchoalveolar lavage fluid total protein, neutrophil counts and percentages, cytokine (KC [CXCl1, mouse IL-8 equivalent] and TNF-α) expression, and activation of NF-κB in lung tissue. Treatment with aerosolized BIO-11006 at 0, 4, 12, 24, and even 36 hours after LPS instillation reversed the disease process: mouse behavior returned to normal after two treatments 12 hours apart with the inhaled peptide after LPS injury, whereas control LPS-instilled animals treated with PBS only remained moribund. Histological appearance of inflammation, bronchoalveolar lavage fluid protein levels, leukocyte and neutrophil numbers, KC and TNF-α gene and protein expression, and NF-κB activation were all significantly attenuated by inhaled BIO-11006 at all time points. These results implicate MARCKS protein in the pathogenesis of ALI/ARDS and suggest that MARCKS-inhibitory peptide(s), delivered by inhalation, could represent a new and potent therapeutic treatment for ALI/ARDS, even if administered well after the disease process has begun.}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Yin, Qi and Fang, Shijing and Park, Joungjoa and Crews, Anne L. and Parikh, Indu and Adler, Kenneth B.}, year={2016}, month={Nov}, pages={617–622} }
 @article{muthusamy_sommerville_moeser_stumpo_sannes_adler_blackshear_weimer_ghashghaei_2015, title={MARCKS-dependent mucin clearance and lipid metabolism in ependymal cells are required for maintenance of forebrain homeostasis during aging}, volume={14}, ISSN={["1474-9726"]}, DOI={10.1111/acel.12354}, abstractNote={Summary}, number={5}, journal={AGING CELL}, author={Muthusamy, Nagendran and Sommerville, Laura J. and Moeser, Adam J. and Stumpo, Deborah J. and Sannes, Philip and Adler, Kenneth and Blackshear, Perry J. and Weimer, Jill M. and Ghashghaei, H. Troy}, year={2015}, month={Oct}, pages={764–773} }
 @misc{chen_chiu_adler_wu_2014, title={A Novel Predictor of Cancer Malignancy: Up-regulation of Myristoylated Alanine-Rich C Kinase Substrate Phosphorylation in Lung Cancer}, volume={189}, ISSN={["1535-4970"]}, DOI={10.1164/rccm.201401-0053le}, abstractNote={To the Editor: 
 
Lung cancer currently remains the leading cause of cancer-related deaths because of its aggressive nature. The 5-year survival rates for localized and regional disease are 54 and 26%, respectively, but only 4% for patients with late-stage (stage IV) disease (1). Thus, development of biomarkers to identify patients at high risk for aggressive progression is of urgent need. Recently, we have reported myristoylated alanine-rich C kinase substrate (MARCKS), predominantly its phosphorylated state, as a risk factor associated with lung cancer invasiveness and metastasis (2). MARCKS is a substrate of protein kinase C, and also a membrane-associated protein. Upon phosphorylation at Ser159 and Ser163 within its phosphorylation site domain, phosphorylated MARCKS (phospho-MARCKS) is detached from the plasma membrane and is able to regulate various cellular processes, including cell migration and exocytic vesicle release (2–4). In the lungs, MARCKS has been extensively studied because of its role in regulating mucus secretion and inflammation. Inhibition of MARCKS activity not only reduces airway mucus hypersecretion both in vitro and in vivo (3, 5), but also represses inflammatory leukocyte migration and degranulation (6, 7). There have been limited studies on MARCKS in cancer metastasis, but the results have been conflicting (8–13). This is because MARCKS expression is ubiquitous in various normal and tumor tissues. Despite this, there is a consensus that phospho-MARCKS, a post-translational modification, is associated with cell motility, and has a role in the regulation of cancer cell invasiveness and metastasis (2, 4, 14, 15). Of note, our laboratory discovered that inhibition of MARCKS phosphorylation was able to reduce lung cancer metastasis in murine models (2). However, the clinical significance of phospho-MARCKS in different cancers remains to be determined. In particular, there is limited information regarding its relevance in cancer progression, especially lung cancer. 
 
Based on 18 pairs of normal and malignant lung cancer tissue sections, we previously reported that elevated phospho-MARCKS was seen in malignant lung cancer tissue sections, but not in their adjacent normal counterparts (2), suggesting a potential association between MARCKS phosphorylation and more aggressive lung cancer histological grades. To investigate more fully this previous finding, we analyzed samples from a cohort of 110 human patients with lung cancer using immunohistochemical staining with an anti-pSer159/163 MARCKS monoclonal antibody (see the online supplement). The clinical characteristics of these patients are summarized in Table 1. Consistent with our previous reports (2), high levels of MARCKS phosphorylation were found in tumor tissues compared with normal lung tissues (Figures 1A–1F). Weak phospho-MARCKS staining was observed in the cytoplasm of lung cancer tissue samples from patients in stage I (Figure 1C). In contrast, strong MARCKS phosphorylation occurred in advanced-stage lung cancer tissue samples (Figures 1D–1F). The levels of MARCKS phosphorylation correlated significantly with advanced stages of disease (Figure 1G, Pearson’s chi-square test). 
 
 
 
Table 1: 
 
Phosphorylated Myristoylated Alanine-Rich C Kinase Substrate Levels in Relation to Clinicopathologic Characteristics of Patients with Non–Small Cell Lung Cancer* 
 
 
 
 
 
Figure 1. 
 
High phosphorylated myristoylated alanine-rich C kinase substrate (phospho-MARCKS) levels correlate with advanced stages, lymph node metastasis, and invasion of lung cancer. (A–F) Representative images of immunohistochemical staining using anti-pSer159/163 ... 
 
 
 
To quantitatively investigate these impressions, bivariate logistic regression models to predict the likelihood of high phospho-MARCKS levels from advanced tumor stages were estimated, and the probabilities of high phospho-MARCKS with stage I to III are shown in Figure 1H. The analyses demonstrated that, for a one-unit increase in stages II and III, the log odds of high expression of phospho-MARCKS levels increased by 1.00 and 2.46 compared with stage I. There were significant differences in the logistic probabilities of high phospho-MARCKS levels between stages I and II (P = 0.039), as well as stages I and III (P < 0.001), respectively. These results suggest that phospho-MARCKS may be a promising clinical predictor of tumor stages in patients with lung cancer. 
 
Moreover, we also investigated the significance of phospho-MARCKS in lymph node status and found that higher levels of MARCKS phosphorylation correlated with lymph node metastasis (Figure 1I, N0 versus N1–2). Notably, MARCKS phosphorylation was lower in a subtype of adenocarcinoma, bronchoalveolar carcinoma, which shows a less invasive phenotype than adenocarcinoma (Figure 1J, AC versus bronchoalveolar carcinoma). Because tumor necrosis is a common event in aggressive cancers, we further checked phospho-MARCKS levels in the 10 tumor tissues with necrosis in this set of tissue arrays. Interestingly, we found higher staining intensity and increased numbers of cells stained with anti–phospho-MARCKS antibody in these tumors. These data raise the possibility that high phospho-MARCKS levels may contribute to cancer progression in non–small cell lung cancers, and the detection of phospho-MARCKS could potentially be used as a prognostic biomarker for the disease. 
 
In this correspondence, we demonstrate that higher MARCKS phosphorylation is correlated with lung cancer in advanced stages (stage II–IV), lymph node metastatic status, and malignant phenotypes. In addition to our previously published results (2), the current work further confirms the importance of phospho-MARCKS in driving the progression of lung cancer toward more malignancy, suggesting that phospho-MARCKS levels may determine the progression of localized lung cancer toward late stage. Taken together, high phospho-MARCKS levels appear to confer cancer malignancy, and may serve as a novel biomarker. Inhibition of MARCKS phosphorylation, the post-translational step, may be an effective strategy for controlling lung cancer progression.}, number={8}, journal={AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE}, author={Chen, Ching-Hsien and Chiu, Chun-Lung and Adler, Kenneth B. and Wu, Reen}, year={2014}, month={Apr}, pages={1002–1004} }
 @article{chen_thai_yoneda_adler_yang_wu_2014, title={A peptide that inhibits function of Myristoylated Alanine-Rich C Kinase Substrate (MARCKS) reduces lung cancer metastasis}, volume={33}, DOI={10.1038/onc.2013.336}, abstractNote={Myristoylated Alanine-Rich C Kinase Substrate (MARCKS), a substrate of protein kinase C, is a key regulatory molecule controlling mucus granule secretion by airway epithelial cells as well as directed migration of leukocytes, stem cells and fibroblasts. Phosphorylation of MARKCS may be involved in these responses. However, the functionality of MARCKS and its related phosphorylation in lung cancer malignancy have not been characterized. This study demonstrated elevated levels of MARCKS and phospho-MARCKS in highly invasive lung cancer cell lines and lung cancer specimens from non-small-cell lung cancer patients. siRNA knockdown of MARCKS expression in these highly invasive lung cancer cell lines reduced cell migration and suppressed PI3K (phosphatidylinositol 3'-kinase)/Akt phosphorylation and Slug level. Interestingly, treatment with a peptide identical to the MARCKS N-terminus sequence (the MANS peptide) impaired cell migration in vitro and also the metastatic potential of invasive lung cancer cells in vivo. Mechanistically, MANS peptide treatment resulted in a coordination of increase of E-cadherin expression, suppression of MARCKS phosphorylation and AKT/Slug signalling pathway but not the expression of total MARCKS. These results indicate a crucial role for MARCKS, specifically its phosphorylated form, in potentiating lung cancer cell migration/metastasis and suggest a potential use of MARCKS-related peptides in the treatment of lung cancer metastasis.}, number={28}, journal={Oncogene}, author={Chen, C. H. and Thai, P. and Yoneda, K. and Adler, K. B. and Yang, P. C. and Wu, R.}, year={2014}, pages={3696–3706} }
 @article{parikh_long_yin_fang_adler_2014, title={Aerosolized MANS and BIO-11006 Peptides Inhibit Lung Cancer Metastasis in SCID Mice}, volume={90}, ISSN={0360-3016}, url={http://dx.doi.org/10.1016/J.IJROBP.2014.08.247}, DOI={10.1016/J.IJROBP.2014.08.247}, abstractNote={Purpose/Objective(s)MARCKS, a ubiquitous 332-aa protein with a highly conserved N-terminal sequence, plays a key role in cell migration. MARCKS elevations in cancer are associated with metastasis and worse outcomes; Chen et al found high phospho MARCKS levels in human NSCLC tissues and showed that MANS, a 24-aa N-terminal MARCKS fragment that inhibits MARCKS function, prevents metastasis of PC9 cells orthotopically injected into the left lung of SCID mice by q3d ip injections. In this study we compared anti-metastatic effects of aerosolized inhaled MANS and its much shorter analog, BIO-11006, in NSCLC in vivo.Materials/MethodsForty SCID mice (n=8 per group) were inoculated with 2.5 x 106 A549 cells by tail vein injection, and treated with aerosolized (Aerogen preclinical nebulizer) MANS, BIO-11006 peptide, or PBS qod for 7 weeks. For each treatment, MANS and BIO-11006 peptide (100 μM), dissolved in five mL PBS, were nebulized over 30 min. The treatment, given every other day, was started either at Day -1, or at Day +3. On Day 53, animals were sacrificed and necropsied. Lungs were harvested, weighed, perfused with 15% India ink, washed with Fekete’s solution, photographed, and metastatic tumor nodules counted and recorded. Other organs were also harvested from control and treated groups and examined for metastasis.ResultsBoth aerosolized MANS and BIO-11006 significantly reduced lung metastasis compared to vehicle control. After the seven week treatment period, the vehicle control exhibited 97±21 metastatic nodules while BIO-11006 (-1 day and +3 day) groups had 34±14* and 21±6**, respectively. Similarly, MANS (-1 day and +3 day) groups showed 22±7* and 13±4** metastatic nodules, respectively. [*=P<.05 and **=P<.01 compared to vehicle; 1-way ANOVA/Bonferroni's Multiple Comparison Test]. Reductions in metastasis caused by MANS and BIO-11006 were not different if aerosol treatment was started before versus after A549 cell injections, and were not different from each other. Two animals exhibited metastasis beyond the lungs; both were in vehicle control group.ConclusionsBoth MARCKS inhibitory peptides, MANS and BIO-11006 significantly inhibit metastasis of A549 cells in SCID mice when administered by aerosol whether treatment is initiated before or after tail vein injection of human A549 lung cancer cells. Purpose/Objective(s)MARCKS, a ubiquitous 332-aa protein with a highly conserved N-terminal sequence, plays a key role in cell migration. MARCKS elevations in cancer are associated with metastasis and worse outcomes; Chen et al found high phospho MARCKS levels in human NSCLC tissues and showed that MANS, a 24-aa N-terminal MARCKS fragment that inhibits MARCKS function, prevents metastasis of PC9 cells orthotopically injected into the left lung of SCID mice by q3d ip injections. In this study we compared anti-metastatic effects of aerosolized inhaled MANS and its much shorter analog, BIO-11006, in NSCLC in vivo. MARCKS, a ubiquitous 332-aa protein with a highly conserved N-terminal sequence, plays a key role in cell migration. MARCKS elevations in cancer are associated with metastasis and worse outcomes; Chen et al found high phospho MARCKS levels in human NSCLC tissues and showed that MANS, a 24-aa N-terminal MARCKS fragment that inhibits MARCKS function, prevents metastasis of PC9 cells orthotopically injected into the left lung of SCID mice by q3d ip injections. In this study we compared anti-metastatic effects of aerosolized inhaled MANS and its much shorter analog, BIO-11006, in NSCLC in vivo. Materials/MethodsForty SCID mice (n=8 per group) were inoculated with 2.5 x 106 A549 cells by tail vein injection, and treated with aerosolized (Aerogen preclinical nebulizer) MANS, BIO-11006 peptide, or PBS qod for 7 weeks. For each treatment, MANS and BIO-11006 peptide (100 μM), dissolved in five mL PBS, were nebulized over 30 min. The treatment, given every other day, was started either at Day -1, or at Day +3. On Day 53, animals were sacrificed and necropsied. Lungs were harvested, weighed, perfused with 15% India ink, washed with Fekete’s solution, photographed, and metastatic tumor nodules counted and recorded. Other organs were also harvested from control and treated groups and examined for metastasis. Forty SCID mice (n=8 per group) were inoculated with 2.5 x 106 A549 cells by tail vein injection, and treated with aerosolized (Aerogen preclinical nebulizer) MANS, BIO-11006 peptide, or PBS qod for 7 weeks. For each treatment, MANS and BIO-11006 peptide (100 μM), dissolved in five mL PBS, were nebulized over 30 min. The treatment, given every other day, was started either at Day -1, or at Day +3. On Day 53, animals were sacrificed and necropsied. Lungs were harvested, weighed, perfused with 15% India ink, washed with Fekete’s solution, photographed, and metastatic tumor nodules counted and recorded. Other organs were also harvested from control and treated groups and examined for metastasis. ResultsBoth aerosolized MANS and BIO-11006 significantly reduced lung metastasis compared to vehicle control. After the seven week treatment period, the vehicle control exhibited 97±21 metastatic nodules while BIO-11006 (-1 day and +3 day) groups had 34±14* and 21±6**, respectively. Similarly, MANS (-1 day and +3 day) groups showed 22±7* and 13±4** metastatic nodules, respectively. [*=P<.05 and **=P<.01 compared to vehicle; 1-way ANOVA/Bonferroni's Multiple Comparison Test]. Reductions in metastasis caused by MANS and BIO-11006 were not different if aerosol treatment was started before versus after A549 cell injections, and were not different from each other. Two animals exhibited metastasis beyond the lungs; both were in vehicle control group. Both aerosolized MANS and BIO-11006 significantly reduced lung metastasis compared to vehicle control. After the seven week treatment period, the vehicle control exhibited 97±21 metastatic nodules while BIO-11006 (-1 day and +3 day) groups had 34±14* and 21±6**, respectively. Similarly, MANS (-1 day and +3 day) groups showed 22±7* and 13±4** metastatic nodules, respectively. [*=P<.05 and **=P<.01 compared to vehicle; 1-way ANOVA/Bonferroni's Multiple Comparison Test]. Reductions in metastasis caused by MANS and BIO-11006 were not different if aerosol treatment was started before versus after A549 cell injections, and were not different from each other. Two animals exhibited metastasis beyond the lungs; both were in vehicle control group. ConclusionsBoth MARCKS inhibitory peptides, MANS and BIO-11006 significantly inhibit metastasis of A549 cells in SCID mice when administered by aerosol whether treatment is initiated before or after tail vein injection of human A549 lung cancer cells. Both MARCKS inhibitory peptides, MANS and BIO-11006 significantly inhibit metastasis of A549 cells in SCID mice when administered by aerosol whether treatment is initiated before or after tail vein injection of human A549 lung cancer cells.}, number={5}, journal={International Journal of Radiation Oncology*Biology*Physics}, publisher={Elsevier BV}, author={Parikh, I. and Long, W.A. and Yin, Q. and Fang, S. and Adler, K.B.}, year={2014}, month={Nov}, pages={S51} }
 @article{sheats_sung_adler_jones_inflammation_2014, title={In Vitro Neutrophil Migration Requires Protein Kinase C-Delta (δ-PKC)-Mediated Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) Phosphorylation}, volume={38}, ISSN={0360-3997 1573-2576}, url={http://dx.doi.org/10.1007/S10753-014-0078-9}, DOI={10.1007/S10753-014-0078-9}, abstractNote={Dysregulated release of neutrophil reactive oxygen species and proteolytic enzymes contributes to both acute and chronic inflammatory diseases. Therefore, molecular regulators of these processes are potential targets for new anti-inflammatory therapies. We have shown previously that myristoylated alanine-rich C-kinase substrate (MARCKS), a well-known actin binding protein and protein kinase C (PKC) substrate, is a key regulator of neutrophil functions. In the current study, we investigate the role of PKC-mediated MARCKS phosphorylation in neutrophil migration and adhesion in vitro. We report that treatment of human neutrophils with the δ-PKC inhibitor rottlerin significantly attenuates f-Met-Leu-Phe (fMLF)-induced MARCKS phosphorylation (IC50 = 5.709 μM), adhesion (IC50 = 8.4 μM), and migration (IC50 = 6.7 μM), while α-, β-, and ζ-PKC inhibitors had no significant effect. We conclude that δ-PKC-mediated MARCKS phosphorylation is essential for human neutrophil migration and adhesion in vitro. These results implicate δ-PKC-mediated MARCKS phosphorylation as a key step in the inflammatory response of neutrophils.}, number={3}, journal={Inflammation}, publisher={Springer Science and Business Media LLC}, author={Sheats, M.K. and Sung, E.J. and Adler, K.B. and Jones, S.L. and Inflammation}, year={2014}, month={Dec}, pages={1126–1141} }
 @article{sheats_pescosolido_hefner_sung_adler_jones_2014, title={Myristoylated Alanine Rich C Kinase Substrate (MARCKS) is essential to β2-integrin dependent responses of equine neutrophils}, volume={160}, ISSN={0165-2427}, url={http://dx.doi.org/10.1016/j.vetimm.2014.04.009}, DOI={10.1016/j.vetimm.2014.04.009}, abstractNote={Neutrophil infiltration is a prominent feature in a number of pathologic conditions affecting horses including recurrent airway obstruction, ischemia-reperfusion injury, and laminitis. Cell signaling components involved in neutrophil migration represent targets for novel anti-inflammatory therapies. In order to migrate into tissue, neutrophils must respond to chemoattractant signals in their external environment through activation of adhesion receptors (i.e. integrins) and reorganization of the actin cytoskeleton. Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS), a highly conserved actin-binding protein, has a well demonstrated role in cytoskeletal dependent cellular functions (i.e. adhesion, spreading, and migration), but the details of MARCKS involvement in these processes remain vague. We hypothesized that MARCKS serves as a link between the actin cytoskeleton and integrin function in neutrophils. Using a MARCKS-specific inhibitor peptide known as MANS on equine neutrophils in vitro, we demonstrate that inhibition of MARCKS function significantly attenuates β2-integrin-dependent neutrophil functions including migration, adhesion, and immune complex-mediated respiratory burst. The MANS peptide did not, however, inhibit the β2-integrin-independent PMA mediated respiratory burst. These results attest to the essential role of MARCKS function in regulating neutrophil responses, and strongly implicate MARCKS as a potential regulator of β2-integrins in neutrophils.}, number={3-4}, journal={Veterinary Immunology and Immunopathology}, publisher={Elsevier BV}, author={Sheats, Mary K. and Pescosolido, Kimberly C. and Hefner, Ethan M. and Sung, Eui Jae and Adler, Kenneth B. and Jones, Samuel L.}, year={2014}, month={Aug}, pages={167–176} }
 @article{chen_statt_chiu_thai_arif_adler_wu_2014, title={Targeting Myristoylated Alanine-Rich C Kinase Substrate Phosphorylation Site Domain in Lung Cancer Mechanisms and Therapeutic Implications}, volume={190}, ISSN={["1535-4970"]}, DOI={10.1164/rccm.201408-1505oc}, abstractNote={RATIONALE
Phosphorylation of myristoylated alanine-rich C kinase substrate (phospho-MARCKS) at the phosphorylation site domain (PSD) is crucial for mucus granule secretion and cell motility, but little is known concerning its function in lung cancer.


OBJECTIVES
We aimed to determine if MARCKS PSD activity can serve as a therapeutic target and to elucidate the molecular basis of this potential.


METHODS
The clinical relevance of phospho-MARCKS was first confirmed. Next, we used genetic approaches to verify the functionality and molecular mechanism of phospho-MARCKS. Finally, cancer cells were pharmacologically inhibited for MARCKS activity and subjected to functional bioassays.


MEASUREMENTS AND MAIN RESULTS
We demonstrated that higher phospho-MARCKS levels were correlated with shorter overall survival of lung cancer patients. Using shRNA silencing and ectopic expression of wild-type and PSD-mutated (S159/163A) MARCKS, we showed that elevated phospho-MARCKS promoted cancer growth and erlotinib resistance. Further studies demonstrated an interaction of phosphoinositide 3-kinase with MARCKS, but not with phospho-MARCKS. Interestingly, phospho-MARCKS acted in parallel with increased phosphatidylinositol (3,4,5)-triphosphate pools and AKT activation in cells. Through treatment with a 25-mer peptide targeting the MARCKS PSD motif (MPS peptide), we were able to suppress tumor growth and metastasis in vivo, and reduced levels of phospho-MARCKS, phosphatidylinositol (3,4,5)-triphosphate, and AKT activity. This peptide also enhanced the sensitivity of lung cancer cells to erlotinib treatment, especially those with sustained activation of phosphoinositide 3-kinase/AKT signaling.


CONCLUSIONS
These results suggest a key role for MARCKS PSD in cancer disease and provide a unique strategy for inhibiting the activity of MARCKS PSD as a treatment for lung cancer.}, number={10}, journal={AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE}, author={Chen, Ching-Hsien and Statt, Sarah and Chiu, Chun-Lung and Thai, Philip and Arif, Muhammad and Adler, Kenneth B. and Wu, Reen}, year={2014}, month={Nov}, pages={1127–1138} }
 @article{adler_shapiro_holtzman_mcdonald_2014, title={The Red Journal at 25 Years Looking Back and Looking Ahead}, volume={50}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2014-0104ed}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Adler, Kenneth B. and Shapiro, Steven D. and Holtzman, Michael J. and McDonald, John A.}, year={2014}, month={May}, pages={839–840} }
 @article{li_d’annibale-tolhurst_adler_fang_yin_birkenheuer_levy_jones_sung_hawkins_et al._2013, title={A Myristoylated Alanine-Rich C Kinase Substrate–Related Peptide Suppresses Cytokine mRNA and Protein Expression in LPS-Activated Canine Neutrophils}, volume={48}, ISSN={1044-1549 1535-4989}, url={http://dx.doi.org/10.1165/rcmb.2012-0278OC}, DOI={10.1165/rcmb.2012-0278oc}, abstractNote={Myristoylated alanine-rich C kinase substrate (MARCKS) is a ubiquitously expressed protein kinase C substrate that has emerged as a potential therapeutic target for the amelioration of mucin secretion and inflammation in patients with chronic obstructive pulmonary disease. MARCKS also plays a key role in regulating the adhesion, migration, and degranulation of neutrophils. Moreover, given its biological role in epithelial and immune cells, we hypothesized that MARCKS may play an integral role in cytokine secretion by neutrophils. Because the amino terminus of MARCKS is highly conserved across vertebrate species, we successfully applied the well-characterized human MARCKS inhibitory peptide, myristoylated N-terminal sequence (MANS), to attenuate the function of MARCKS in isolated canine neutrophils. Pretreatment of canine neutrophils with MANS peptide significantly reduced both mRNA and protein expression in a broad range of LPS-induced cytokines, including IL-8, a chemokine (C-X-C motif) ligand-1 orthologue, and TNF-α, in comparison with untreated cells or those treated with a control peptide. This reduction in cytokine expression was observed even when neutrophils were treated with MANS 2 hours after LPS exposure. The observed reduction in cytokine secretion was not attributable to protein retention or cell death, but was associated with reduced cytokine transcript synthesis. These observations identify MARCKS protein as a promising therapeutic target in the treatment of inflammatory diseases or syndromes attributed to neutrophil influx and inflammatory cytokine production, such as sepsis, acute lung injury, and acute respiratory distress syndrome.}, number={3}, journal={American Journal of Respiratory Cell and Molecular Biology}, publisher={American Thoracic Society}, author={Li, Jingjing and D’Annibale-Tolhurst, Melissa A. and Adler, Kenneth B. and Fang, Shijing and Yin, Qui and Birkenheuer, Adam J. and Levy, Michael G. and Jones, Samuel L. and Sung, Eui Jae and Hawkins, Eleanor C. and et al.}, year={2013}, month={Mar}, pages={314–321} }
 @article{ott_sung_melvin_sheats_haugh_adler_jones_2013, title={Fibroblast Migration Is Regulated by Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) Protein}, volume={8}, ISSN={["1932-6203"]}, url={http://europepmc.org/abstract/med/23840497}, DOI={10.1371/journal.pone.0066512}, abstractNote={Myristoylated alanine-rich C-kinase substrate (MARCKS) is a ubiquitously expressed substrate of protein kinase C (PKC) that is involved in reorganization of the actin cytoskeleton. We hypothesized that MARCKS is involved in regulation of fibroblast migration and addressed this hypothesis by utilizing a unique reagent developed in this laboratory, the MANS peptide. The MANS peptide is a myristoylated cell permeable peptide corresponding to the first 24-amino acids of MARCKS that inhibits MARCKS function. Treatment of NIH-3T3 fibroblasts with the MANS peptide attenuated cell migration in scratch wounding assays, while a myristoylated, missense control peptide (RNS) had no effect. Neither MANS nor RNS peptide treatment altered NIH-3T3 cell proliferation within the parameters of the scratch assay. MANS peptide treatment also resulted in inhibited NIH-3T3 chemotaxis towards the chemoattractant platelet-derived growth factor-BB (PDGF-BB), with no effect observed with RNS treatment. Live cell imaging of PDGF-BB induced chemotaxis demonstrated that MANS peptide treatment resulted in weak chemotactic fidelity compared to RNS treated cells. MANS and RNS peptides did not affect PDGF-BB induced phosphorylation of MARCKS or phosphoinositide 3-kinase (PI3K) signaling, as measured by Akt phosphorylation. Further, no difference in cell migration was observed in NIH-3T3 fibroblasts that were transfected with MARCKS siRNAs with or without MANS peptide treatment. Genetic structure-function analysis revealed that MANS peptide-mediated attenuation of NIH-3T3 cell migration does not require the presence of the myristic acid moiety on the amino-terminus. Expression of either MANS or unmyristoylated MANS (UMANS) C-terminal EGFP fusion proteins resulted in similar levels of attenuated cell migration as observed with MANS peptide treatment. These data demonstrate that MARCKS regulates cell migration and suggests that MARCKS-mediated regulation of fibroblast migration involves the MARCKS amino-terminus. Further, this data demonstrates that MANS peptide treatment inhibits MARCKS function during fibroblast migration and that MANS mediated inhibition occurs independent of myristoylation.}, number={6}, journal={PLOS ONE}, publisher={Public Library of Science (PLoS)}, author={Ott, Laura E. and Sung, Eui Jae and Melvin, Adam T. and Sheats, Mary K. and Haugh, Jason M. and Adler, Kenneth B. and Jones, Samuel L.}, editor={Aspenstrom, PontusEditor}, year={2013}, month={Jun} }
 @article{newcomb_boswell_sherrill_polosukhin_boyd_goleniewska_brody_kolls_adler_peebles_2013, title={IL-17A Induces Signal Transducers and Activators of Transcription-6-Independent Airway Mucous Cell Metaplasia}, volume={48}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2013-0017oc}, abstractNote={Mucous cell metaplasia is a hallmark of asthma, and may be mediated by signal transducers and activators of transcription (STAT)-6 signaling. IL-17A is increased in the bronchoalveolar lavage fluid of patients with severe asthma, and IL-17A also increases mucus production in airway epithelial cells. Asthma therapeutics are being developed that inhibit STAT6 signaling, but the role of IL-17A in inducing mucus production in the absence of STAT6 remains unknown. We hypothesized that IL-17A induces mucous cell metaplasia independent of STAT6, and we tested this hypothesis in two murine models in which increased IL-17A protein expression is evident. In the first model, ovalbumin (OVA)-specific D011.10 Th17 cells were adoptively transferred into wild-type (WT) or STAT6 knockout (KO) mice, and the mice were challenged with OVA or PBS. WT-OVA and STAT6 KO-OVA mice demonstrated increased airway IL-17A and IL-13 protein expression and mucous cell metaplasia, compared with WT-PBS or STAT6 KO-PBS mice. In the second model, WT, STAT1 KO, STAT1/STAT6 double KO (DKO), or STAT1/STAT6/IL-17 receptor A (RA) triple KO (TKO) mice were challenged with respiratory syncytial virus (RSV) or mock viral preparation, and the mucous cells were assessed. STAT1 KO-RSV mice demonstrated increased airway mucous cell metaplasia compared with WT-RSV mice. STAT1 KO-RSV and STAT1/STAT6 DKO-RSV mice also demonstrated increased mucous cell metaplasia, compared with STAT1/STAT6/IL17RA TKO-RSV mice. We also treated primary murine tracheal epithelial cells (mTECs) from WT and STAT6 KO mice. STAT6 KO mTECs showed increased periodic acid-Schiff staining with IL-17A but not with IL-13. Thus, asthma therapies targeting STAT6 may increase IL-17A protein expression, without preventing IL-17A-induced mucus production.}, number={6}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Newcomb, Dawn C. and Boswell, Madison G. and Sherrill, Taylor P. and Polosukhin, Vasiliy V. and Boyd, Kelli L. and Goleniewska, Kasia and Brody, Steven L. and Kolls, Jay K. and Adler, Kenneth B. and Peebles, Stokes, Jr.}, year={2013}, month={Jun}, pages={711–716} }
 @article{fang_crews_chen_park_yin_ren_adler_2013, title={MARCKS and HSP70 interactions regulate mucin secretion by human airway epithelial cells in vitro}, volume={304}, ISSN={["1522-1504"]}, DOI={10.1152/ajplung.00337.2012}, abstractNote={Myristoylated alanine-rich C kinase substrate (MARCKS) protein has been recognized as a key regulatory molecule controlling mucin secretion by airway epithelial cells in vitro and in vivo. We recently showed that two intracellular chaperones, heat shock protein 70 (HSP70) and cysteine string protein (CSP), associate with MARCKS in the secretory mechanism. To elucidate more fully MARCKS-HSP70 interactions in this process, studies were performed in well-differentiated normal human bronchial epithelial (NHBE) cells maintained in air-liquid interface culture utilizing specific pharmacological inhibition of HSP70 with pyrimidinone MAL3-101 and siRNA approaches. The results indicate that HSP70 interaction with MARCKS is enhanced after exposure of the cells to the protein kinase C activator/mucin secretagogue, phorbol 12-myristate 13-acetate (PMA). Pretreatment of NHBEs with MAL3-101 attenuated in a concentration-dependent manner PMA-stimulated mucin secretion and interactions among HSP70, MARCKS, and CSP. In additional studies, trafficking of MARCKS in living NHBE cells was investigated after transfecting cells with fluorescently tagged DNA constructs: MARCKS-yellow fluorescent protein, and/or HSP70-cyan fluorescent protein. Cells were treated with PMA 48 h posttransfection, and trafficking of the constructs was examined by confocal microscopy. MARCKS translocated rapidly from plasma membrane to cytoplasm, whereas HSP70 was observed in the cytoplasm and appeared to associate with MARCKS after PMA exposure. Pretreatment of cells with either MAL3-101 or HSP70 siRNA inhibited translocation of MARCKS. These results provide evidence of a role for HSP70 in mediating mucin secretion via interactions with MARCKS and that these interactions are critical for the cytoplasmic translocation of MARCKS upon its phosphorylation.}, number={8}, journal={AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY}, author={Fang, Shijing and Crews, Anne L. and Chen, Wei and Park, Joungjoa and Yin, Qi and Ren, Xiu-Rong and Adler, Kenneth B.}, year={2013}, month={Apr}, pages={L511–L518} }
 @article{lampe_park_fang_crews_adler_2012, title={Calpain and MARCKS protein regulation of airway mucin secretion}, volume={25}, ISSN={["1094-5539"]}, DOI={10.1016/j.pupt.2012.06.003}, abstractNote={Hypersecretion of mucin plays an important role in the pathophysiology of many inflammatory airway diseases, including asthma, chronic bronchitis, and cystic fibrosis. Myristoylated alanine-rich C-kinase substrate (MARCKS) protein has been shown to play an important role in regulation of airway mucin secretion, as peptides analogous to the amino (N)-terminus of MARCKS attenuate mucin secretion by airway epithelium in vitro and in vivo. Here, we investigated a potential role for the protease Calpain, a calcium-dependent cysteine protease that can cleave MARCKS, in the MARCKS-related secretory mechanism. We theorized that Calpain might cleave MARCKS near the N-terminus, thereby attenuating the ability of MARCKS to bind to membranes and/or creating a small N-terminal peptide that could act as a competitive intracellular inhibitor to remaining endogenous full-length MARCKS molecules. Primary normal human bronchial epithelial (NHBE) cells and the virally-transformed human bronchial epithelial HBE1 cell line were exposed to phorbol-12-myristate-13-acetate (PMA) to stimulate the Protein Kinase C (PKC) pathway, leading to enhanced mucin secretion, and Calpain activity within the cells was measured with a fluorescent cleavage assay. Calpain activity was increased by PMA, and pretreatment of the cells with Calpain inhibitors reduced both Calpain activity and mucin secretion in a concentration-dependent manner. Thus, as opposed to the original hypothesis, inactivating Calpain caused a decrease rather than an increase in secretion. HBE1 cells transfected with DNA constructs encoding a MARCKS-YFP fusion protein showed cleavage at a putative site near the N-terminus in response to PMA. Cleavage of MARCKS by Calpain may have an important role in regulation of the PKC/MARCKS pathway regulating airway mucin secretion.}, number={6}, journal={PULMONARY PHARMACOLOGY & THERAPEUTICS}, author={Lampe, W. Randall and Park, Joungjoa and Fang, Shijing and Crews, Anne L. and Adler, Kenneth B.}, year={2012}, month={Dec}, pages={427–431} }
 @article{green_park_yin_fang_crews_jones_adler_2012, title={Directed migration of mouse macrophages in vitro involves myristoylated alanine-rich C-kinase substrate (MARCKS) protein}, volume={92}, ISSN={0741-5400}, url={http://dx.doi.org/10.1189/jlb.1211604}, DOI={10.1189/jlb.1211604}, abstractNote={Abstract}, number={3}, journal={Journal of Leukocyte Biology}, publisher={Wiley}, author={Green, T. D. and Park, J. and Yin, Q. and Fang, S. and Crews, A. L. and Jones, S. L. and Adler, K. B.}, year={2012}, month={May}, pages={633–639} }
 @article{boswell_newcomb_goleniewska_reiss_adler_peebles_2012, title={IL-17A-mediated STAT6 Independent Mucus Production in Mice}, volume={129}, ISSN={0091-6749}, url={http://dx.doi.org/10.1016/j.jaci.2011.12.308}, DOI={10.1016/j.jaci.2011.12.308}, abstractNote={RationaleThe prevailing current concept is that airway mucus expression is dependent on STAT6 signaling. However, IL-17A has recently been reported to induce airway mucus expression and is not known to signal through STAT6. We hypothesized that IL-17A induces airway mucus in a STAT6-independent manner. To test this hypothesis, we used a mouse model of RSV infection of STAT1 knock out (KO) mice in which there is abundant lung IL-17A protein expression. We created STAT1/STAT6 double KO (DKO) mice to determine if the mucus in the RSV-infected STAT1 KO mice was dependent upon STAT6 signaling, and STAT1/STAT6/IL-17RA TKO to confirm the role of IL-17A.MethodsBALB/c WT, STAT1 KO, STAT/STAT6 DKO, and STAT1/STAT6/IL-17RA TKO mice were infected with RSV strain A2 and lungs were harvested 6 days post infection and airway mucus expression was assessed by histopathology. Fully differentiated mouse airway epithelial cells (AECs) were cultured from WT and STAT6 KO mice and were cultured with PBS, IL-13, or IL-17 and mucus expression was assessed by histopathology.ResultsSTAT1 KO and STAT1/STAT6 DKO mice had increased lung IL-17A expression and airway mucus expression, but no mucus was detected in STAT1/STAT6/IL-17RA TKO (n=4-7, ANOVA). While AECs from WT mice expressed mucus when cultured with either IL-13 or IL-17A, AECs from STAT6 KO mice expressed mucus only when cultured with IL-17A, but not IL-13.ConclusionsIL-17A induces STAT6-independent airway mucus expression. Asthma therapies targeting the STAT6 signaling pathway will not prevent Il-17A-induced mucus expression and IL-17A has been linked to severe asthma. RationaleThe prevailing current concept is that airway mucus expression is dependent on STAT6 signaling. However, IL-17A has recently been reported to induce airway mucus expression and is not known to signal through STAT6. We hypothesized that IL-17A induces airway mucus in a STAT6-independent manner. To test this hypothesis, we used a mouse model of RSV infection of STAT1 knock out (KO) mice in which there is abundant lung IL-17A protein expression. We created STAT1/STAT6 double KO (DKO) mice to determine if the mucus in the RSV-infected STAT1 KO mice was dependent upon STAT6 signaling, and STAT1/STAT6/IL-17RA TKO to confirm the role of IL-17A. The prevailing current concept is that airway mucus expression is dependent on STAT6 signaling. However, IL-17A has recently been reported to induce airway mucus expression and is not known to signal through STAT6. We hypothesized that IL-17A induces airway mucus in a STAT6-independent manner. To test this hypothesis, we used a mouse model of RSV infection of STAT1 knock out (KO) mice in which there is abundant lung IL-17A protein expression. We created STAT1/STAT6 double KO (DKO) mice to determine if the mucus in the RSV-infected STAT1 KO mice was dependent upon STAT6 signaling, and STAT1/STAT6/IL-17RA TKO to confirm the role of IL-17A. MethodsBALB/c WT, STAT1 KO, STAT/STAT6 DKO, and STAT1/STAT6/IL-17RA TKO mice were infected with RSV strain A2 and lungs were harvested 6 days post infection and airway mucus expression was assessed by histopathology. Fully differentiated mouse airway epithelial cells (AECs) were cultured from WT and STAT6 KO mice and were cultured with PBS, IL-13, or IL-17 and mucus expression was assessed by histopathology. BALB/c WT, STAT1 KO, STAT/STAT6 DKO, and STAT1/STAT6/IL-17RA TKO mice were infected with RSV strain A2 and lungs were harvested 6 days post infection and airway mucus expression was assessed by histopathology. Fully differentiated mouse airway epithelial cells (AECs) were cultured from WT and STAT6 KO mice and were cultured with PBS, IL-13, or IL-17 and mucus expression was assessed by histopathology. ResultsSTAT1 KO and STAT1/STAT6 DKO mice had increased lung IL-17A expression and airway mucus expression, but no mucus was detected in STAT1/STAT6/IL-17RA TKO (n=4-7, ANOVA). While AECs from WT mice expressed mucus when cultured with either IL-13 or IL-17A, AECs from STAT6 KO mice expressed mucus only when cultured with IL-17A, but not IL-13. STAT1 KO and STAT1/STAT6 DKO mice had increased lung IL-17A expression and airway mucus expression, but no mucus was detected in STAT1/STAT6/IL-17RA TKO (n=4-7, ANOVA). While AECs from WT mice expressed mucus when cultured with either IL-13 or IL-17A, AECs from STAT6 KO mice expressed mucus only when cultured with IL-17A, but not IL-13. ConclusionsIL-17A induces STAT6-independent airway mucus expression. Asthma therapies targeting the STAT6 signaling pathway will not prevent Il-17A-induced mucus expression and IL-17A has been linked to severe asthma. IL-17A induces STAT6-independent airway mucus expression. Asthma therapies targeting the STAT6 signaling pathway will not prevent Il-17A-induced mucus expression and IL-17A has been linked to severe asthma.}, number={2}, journal={Journal of Allergy and Clinical Immunology}, publisher={Elsevier BV}, author={Boswell, M.G. and Newcomb, D.C. and Goleniewska, K. and Reiss, S. and Adler, K.B. and Peebles, R.S., Jr}, year={2012}, month={Feb}, pages={AB199} }
 @article{seibold_wise_speer_steele_brown_loyd_fingerlin_zhang_gudmundsson_groshong_et al._2011, title={A Common MUC5B Promoter Polymorphism and Pulmonary Fibrosis}, volume={364}, ISSN={["1533-4406"]}, DOI={10.1056/nejmoa1013660}, abstractNote={BACKGROUND
The mutations that have been implicated in pulmonary fibrosis account for only a small proportion of the population risk.


METHODS
Using a genomewide linkage scan, we detected linkage between idiopathic interstitial pneumonia and a 3.4-Mb region of chromosome 11p15 in 82 families. We then evaluated genetic variation in this region in gel-forming mucin genes expressed in the lung among 83 subjects with familial interstitial pneumonia, 492 subjects with idiopathic pulmonary fibrosis, and 322 controls. MUC5B expression was assessed in lung tissue.


RESULTS
Linkage and fine mapping were used to identify a region of interest on the p-terminus of chromosome 11 that included gel-forming mucin genes. The minor-allele of the single-nucleotide polymorphism (SNP) rs35705950, located 3 kb upstream of the MUC5B transcription start site, was present at a frequency of 34% among subjects with familial interstitial pneumonia, 38% among subjects with idiopathic pulmonary fibrosis, and 9% among controls (allelic association with familial interstitial pneumonia, P=1.2×10(-15); allelic association with idiopathic pulmonary fibrosis, P=2.5×10(-37)). The odds ratios for disease among subjects who were heterozygous and those who were homozygous for the minor allele of this SNP were 6.8 (95% confidence interval [CI], 3.9 to 12.0) and 20.8 (95% CI, 3.8 to 113.7), respectively, for familial interstitial pneumonia and 9.0 (95% CI, 6.2 to 13.1) and 21.8 (95% CI, 5.1 to 93.5), respectively, for idiopathic pulmonary fibrosis. MUC5B expression in the lung was 14.1 times as high in subjects who had idiopathic pulmonary fibrosis as in those who did not (P<0.001). The variant allele of rs35705950 was associated with up-regulation in MUC5B expression in the lung in unaffected subjects (expression was 37.4 times as high as in unaffected subjects homozygous for the wild-type allele, P<0.001). MUC5B protein was expressed in lesions of idiopathic pulmonary fibrosis.


CONCLUSIONS
A common polymorphism in the promoter of MUC5B is associated with familial interstitial pneumonia and idiopathic pulmonary fibrosis. Our findings suggest that dysregulated MUC5B expression in the lung may be involved in the pathogenesis of pulmonary fibrosis. (Funded by the National Heart, Lung, and Blood Institute and others.).}, number={16}, journal={NEW ENGLAND JOURNAL OF MEDICINE}, author={Seibold, Max A. and Wise, Anastasia L. and Speer, Marcy C. and Steele, Mark P. and Brown, Kevin K. and Loyd, James E. and Fingerlin, Tasha E. and Zhang, Weiming and Gudmundsson, Gunnar and Groshong, Steve D. and et al.}, year={2011}, month={Apr}, pages={1503–1512} }
 @article{raiford_park_lin_fang_crews_adler_2011, title={Mucin granule-associated proteins in human bronchial epithelial cells: the airway goblet cell "granulome"}, volume={12}, ISSN={["1465-993X"]}, DOI={10.1186/1465-9921-12-118}, abstractNote={Abstract}, journal={RESPIRATORY RESEARCH}, author={Raiford, Kimberly L. and Park, Joungjoa and Lin, Ko-Wei and Fang, Shijing and Crews, Anne L. and Adler, Kenneth B.}, year={2011}, month={Sep} }
 @misc{green_crews_park_fang_adler_2011, title={Regulation of mucin secretion and inflammation in asthma: A role for MARCKS protein?}, volume={1810}, ISSN={["0304-4165"]}, DOI={10.1016/j.bbagen.2011.01.009}, abstractNote={A major characteristic of asthmatic airways is an increase in mucin (the glycoprotein component of mucus) producing and secreting cells, which leads to increased mucin release that further clogs constricted airways and contributes markedly to airway obstruction and, in the most severe cases, to status asthmaticus. Asthmatic airways show both a hyperplasia and metaplasia of goblet cells, mucin-producing cells in the epithelium; hyperplasia refers to enhanced numbers of goblet cells in larger airways, while metaplasia refers to the appearance of these cells in smaller airways where they normally are not seen. With the number of mucin-producing and secreting cells increased, there is a coincident hypersecretion of mucin which characterizes asthma. On a cellular level, a major regulator of airway mucin secretion in both in vitro and in vivo studies has been shown to be MARCKS (myristoylated alanine-rich C kinase substrate) protein, a ubiquitous substrate of protein kinase C (PKC). In this review, properties of MARCKS and how the protein may regulate mucin secretion at a cellular level will be discussed. In addition, the roles of MARCKS in airway inflammation related to both influx of inflammatory cells into the lung and release of granules containing inflammatory mediators by these cells will be explored. This article is part of a Special Issue entitled: Biochemistry of Asthma.}, number={11}, journal={BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS}, author={Green, Teresa D. and Crews, Anne L. and Park, Joungjoa and Fang, Shijing and Adler, Kenneth B.}, year={2011}, month={Nov}, pages={1110–1113} }
 @article{ott_mcdowell_turner_law_adler_yoder_jones_2011, place={Hoboken, N.J}, title={Two Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) Paralogs are Required for Normal Development in Zebrafish}, volume={294}, ISSN={1932-8486}, url={http://dx.doi.org/10.1002/ar.21453}, DOI={10.1002/ar.21453}, abstractNote={Abstract}, note={: 2007),}, number={9}, journal={The Anatomical Record: Advances in Integrative Anatomy and Evolutionary Biology}, publisher={Wiley}, author={Ott, Laura E. and Mcdowell, Zachary T. and Turner, Poem M. and Law, J. Mchugh and Adler, Kenneth B. and Yoder, Jeffrey A. and Jones, Samuel L.}, year={2011}, month={Aug}, pages={1511–1524} }
 @article{lin_fang_park_crews_adler_2010, title={MARCKS and Related Chaperones Bind to Unconventional Myosin V Isoforms in Airway Epithelial Cells}, volume={43}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2010-0016rc}, abstractNote={We have shown previously that myristoylated alanine-rich C kinase substrate (MARCKS) is a key regulatory molecule in the process of mucin secretion by airway epithelial cells, and that part of the secretory mechanism involves intracellular associations of MARCKS with specific chaperones: heat shock protein 70 (Hsp70) and cysteine string protein (CSP). Here, we report that MARCKS also interacts with unconventional myosin isoforms within these cells, and further molecular interactions between MARCKS and these chaperones/cytoskeletal proteins are elucidated. Primary human bronchial epithelial cells and the HBE1 cell line both expressed myosin V and VI proteins, and both MARCKS and CSP were shown to bind to myosin V, specifically Va and Vc. This binding was enhanced by exposing the cells to phorbol-12-myristate-13-acetate, an activator of protein kinase C and stimulator of mucin secretion. Binding of MARCKS, Hsp70, and CSP was further investigated by His-tagged pull down assays of purified recombinant proteins and multiple transfections of HBE1 cells with fusion proteins (MARCKS-HA; Flag-Hsp70; c-Myc-CSP) and immunoprecipitation. The results showed that MARCKS binds directly to Hsp70, and that Hsp70 binds directly to CSP, but that MARCKS binding to CSP appears to require the presence of Hsp70. Interrelated binding(s) of MARCKS, chaperones, and unconventional myosin isoforms may be integral to the mucin secretion process.}, number={2}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Lin, Ko-Wei and Fang, Shijing and Park, Joungjoa and Crews, Anne L. and Adler, Kenneth B.}, year={2010}, month={Aug}, pages={131–136} }
 @article{foster_adler_crews_potts_fischer_voynow_2010, title={MARCKS-related peptide modulates in vivo the secretion of airway Muc5ac (Retracted article. See vol. 309, pg. L 750, 2015)}, volume={299}, ISSN={["1522-1504"]}, DOI={10.1152/ajplung.00067.2010}, abstractNote={ In a mouse model of neutrophil elastase-induced bronchitis that exhibits goblet cell metaplasia and inflammation, we investigated the effects of intratracheal instillation of the MANS peptide, a peptide identical to the NH2 terminus of the myristoylated alanine-rich C kinase substrate (MARCKS) on mucin protein airway secretion, inflammation, and airway reactivity. To induce mucus cell metaplasia in the airways, male BALB/c mice were treated repetitively with the serine protease, neutrophil elastase, on days 1, 4, and 7. On day 11, when goblet cell metaplasia was fully developed and profiles of proinflammatory cytokines were maximal, the animals were exposed to aerosolized methacholine after intratracheal instillation of MANS or a missense control peptide (RNS). MANS, but not RNS, attenuated the methacholine-stimulated secretion of the major respiratory mucin protein, Muc5ac (50% reduction). Concurrently, elastase-induced proinflammatory cytokines typically recovered in bronchoalveolar lavage (BAL), including KC, IL-1β, IL-6, MCP-1, and TNFα, were reduced by the MANS peptide (mean levels decreased 50–60%). Secondary to the effects of MANS on mucin secretion and inflammation, mechanical lung function by forced oscillation technique was characterized with respect to airway reactivity in response to cumulative aerosol stimulation with serotonin. The MANS peptide was also found to effectively attenuate airway hyperresponsiveness to serotonin in this airway hypersecretory model. Collectively, these findings support the concept that even in airway epithelia remodeled with goblet cell metaplasia and in a state of mucin hypersecretion, exogenous attenuation of function of MARCKS protein via the MANS peptide decreases airway mucin secretion, inflammation, and hyperreactivity. }, number={3}, journal={AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY}, author={Foster, W. Michael and Adler, Kenneth B. and Crews, Anne L. and Potts, Erin N. and Fischer, Bernard M. and Voynow, Judith A.}, year={2010}, month={Sep}, pages={L345–L352} }
 @article{miller_lankford_adler_brody_2010, title={Mesenchymal Stem Cells Require MARCKS Protein for Directed ChemotaxisIn Vitro}, volume={43}, ISSN={1044-1549 1535-4989}, url={http://dx.doi.org/10.1165/rcmb.2010-0015RC}, DOI={10.1165/rcmb.2010-0015rc}, abstractNote={Mesenchymal stem cells (MSCs) reside within tissues such as bone marrow, cord blood, and dental pulp and can differentiate into other mesenchymal cell types. Differentiated MSCs, called circulating fibrocytes, have been demonstrated in human lungs and migrate to injured lung tissue in experimental models. It is likely that MSCs migrate from the bone marrow to sites of injury by following increasing chemokine concentrations. In the present study, we show that primary mouse bone marrow mesenchymal stem cells (BM-MSCs) exhibit directed chemotaxis through transwell inserts toward increasing concentrations of the chemokines complement component 5a, stromal cell-derived factor-1alpha, and monocyte chemotactic protein-1. Prior research has indicated that myristoylated alanine-rich C kinase substrate (MARCKS) protein is critically important for motility in macrophages, neutrophils, and fibroblasts, and here we investigated a possible role for MARCKS in BM-MSC directed chemotaxis. The presence of MARCKS in these cells as well as in human cord blood MSC was verified by Western blotting, and MARCKS was rapidly phosphorylated in these cells after exposure to chemokines. A synthetic peptide that inhibits MARCKS function attenuated, in a concentration-dependent manner, directed chemotaxis of BM-MSCs, while a missense control peptide had no effect. Our results illustrate, for the first time, that MARCKS protein plays an integral role in BM-MSC-directed chemotaxis in vitro.}, number={3}, journal={American Journal of Respiratory Cell and Molecular Biology}, publisher={American Thoracic Society}, author={Miller, Jeffrey D. and Lankford, Susan M. and Adler, Kenneth B. and Brody, Arnold R.}, year={2010}, month={Sep}, pages={253–258} }
 @article{eckert_neuder_park_adler_jones_2010, title={Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS) Protein Regulation of Human Neutrophil Migration}, volume={42}, ISSN={["1535-4989"]}, url={http://europepmc.org/abstract/med/19574534}, DOI={10.1165/rcmb.2008-0394oc}, abstractNote={Neutrophil migration into infected tissues is essential for host defense, but products of activated neutrophils can be quite damaging to host cells. Neutrophil influx into the lung and airways and resultant inflammation characterizes diseases such as chronic obstructive pulmonary disease, bronchiectasis, and cystic fibrosis. To migrate, neutrophils must reorganize the actin cytoskeleton to establish a leading edge pseudopod and a trailing edge uropod. The actin-binding protein myristoylated alanine-rich C-kinase substrate (MARCKS) has been shown to bind and cross-link actin in a variety of cell types and to co-localize with F-actin in the leading edge lamellipodium of migrating fibroblasts. The hypothesis that MARCKS has a role in the regulation of neutrophil migration was tested using a cell-permeant peptide derived from the MARCKS myristoylated aminoterminus (MANS peptide). Treatment of isolated human neutrophils with MANS significantly inhibited both their migration and beta2 integrin-dependent adhesion in response to N-formyl-methionyl-leucyl-phenylalanine (fMLF), IL-8, or leukotriene (LT)B(4). The IC(50) for fMLF-induced migration and adhesion was 17.1 microM and 12.5 microM, respectively. MANS significantly reduced the F-actin content in neutrophils 30 seconds after fMLF stimulation, although the peptide did not alter the ability of cells to polarize or spread. MANS did not alter fMLF-induced increases in surface beta2 integrin expression. These results suggest that MARCKS, via its myristoylated aminoterminus, is a key regulator of neutrophil migration and adhesion.}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Eckert, Rachael E. and Neuder, Laura E. and Park, Joungjoa and Adler, Kenneth B. and Jones, Samuel L.}, year={2010}, month={May}, pages={586–594} }
 @article{adler_abraham_2009, title={Changes in NIH Review Procedures Strengths and Weaknesses}, volume={41}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2009-0201ed}, number={2}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Adler, Kenneth B. and Abraham, Edward}, year={2009}, month={Aug}, pages={127–128} }
 @article{adler_abraham_2009, title={Changes in NIH Review Procedures Strengths and Weaknesses}, volume={180}, ISSN={["1073-449X"]}, DOI={10.1164/rccm.200906-0868ed}, abstractNote={‘‘Change has come’’ to the scoring and review system that all NIH study sections will soon incorporate. The purpose of this editorial is to describe this new system and to offer our own impressions, comments, and concerns. These changes in the review procedures for NIH grant applications are described in the websites for all of the institutes and were summarized in the ‘‘NIH Corner’’ segment on the inside cover of the July American Journal of Respiratory Cell and Molecular Biology. Without going into great detail, the major changes are:}, number={3}, journal={AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE}, author={Adler, Kenneth B. and Abraham, Edward}, year={2009}, month={Aug}, pages={197–197} }
 @article{adler_matalon_2009, title={Highlights of the August issue}, volume={41}, DOI={10.1165/rcmb.2009-2008ed}, number={2}, journal={American Journal of Respiratory Cell and Molecular Biology}, author={Adler, K. B. and Matalon, S.}, year={2009}, pages={125–126} }
 @article{adler_matalon_2009, title={Highlights of the July Issue}, volume={41}, DOI={10.1165/rcmb.2009-2007ed}, number={1}, journal={American Journal of Respiratory Cell and Molecular Biology}, author={Adler, K. B. and Matalon, S.}, year={2009}, pages={1–2} }
 @article{crews_potts_voynow_fischer_bumgardner_adler_foster_2008, title={A MARCKS-related peptide attenuates both mucin hypersecretion and inflammatory cell infiltration in an elastase model of chronic bronchitis.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Crews, A. L. and Potts, E. and Voynow, J. A. and Fischer, B. M. and Bumgardner, S. and Adler, K. B. and Foster, W. M.}, year={2008}, pages={A332} }
 @article{wang_adler_slade_church_webb_chu_jinwright_crews_kraft_2008, title={A MARCKS-related peptide inhibits MUC5AC expression in airway epithelial cells isolated from asthmatic subjects.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Wang, Y. and Adler, K. and Slade, D. J. and Church, T. D. and Webb, R. F. and Chu, H. W. and Jinwright, D. and Crews, A. and Kraft, M.}, year={2008}, pages={A814} }
 @article{park_crews_alper_schwartz_adler_2008, title={A potential role for airway epithelial cell mucin genes in fibrogenesis.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Park, J. J. and Crews, A. L. and Alper, S. and Schwartz, D. A. and Adler, K. B.}, year={2008}, pages={A727} }
 @article{raiford_adler_2008, title={A proteomic analysis of proteins associated with mucin granule membranes within airway epithelial cells.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Raiford, K. L. and Adler, K. B.}, year={2008}, pages={A993} }
 @article{adler_2008, title={After 20 years, what next for the Red journal?}, volume={39}, ISSN={["1044-1549"]}, DOI={10.1165/rcmb.2008-0005ed}, number={4}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Adler, Kenneth B.}, year={2008}, month={Oct}, pages={III-IV} }
 @article{waterman_jones_adler_2008, title={Albuterol-induced oxidant stress in primary cultures of normal human bronchial epithelial cells.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Waterman, J. T. and Jones, J. N. and Adler, K. B.}, year={2008}, pages={A469} }
 @article{wang_adler_2008, title={Epithelial brush border proteomics and associated dysfunction.}, volume={1}, journal={404nOtfound}, author={Wang, X. and Adler, K. B.}, year={2008}, pages={27–33} }
 @article{jester_jiang_zhao_j._i._t._k. b._panettieri_2008, title={Inhibition of ozone-induced airway inflammation by decreasing myristoylated alanine-rich C kinase substrate function.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Jester, W. F. and Jiang, M. and Zhao, H. Liang and J., Parikh and I., Murphy and T., Adler and K. B. and Panettieri, R. A}, year={2008}, pages={A495} }
 @article{lin_j. j._s._adler_2008, title={MARCKS protein regulation of mucin secretion in airway epithelial cells: binding of MARCKS to chaperones and to MyosinV.}, volume={177}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Lin, K. Park and J. J., Fang and S. and Adler, K. B.}, year={2008}, pages={A993} }
 @article{park_fang_crews_lin_adler_2008, title={MARCKS regulation of mucin secretion by airway epithelial cells in vitro: interactions with chaperone proteins.}, volume={102}, journal={American Journal of Respiratory Cell and Molecular Biology}, author={Park, J. J. and Fang, S. and Crews, A. L. and Lin, K.-W. and Adler, K. B.}, year={2008}, pages={949–955} }
 @article{park_fang_crews_lin_adler_2008, title={MARCKS regulation of mucin secretion by airway epithelium in vitro - Interaction with chaperones}, volume={39}, DOI={10.1165/rcm6.2007-0139OC}, number={1}, journal={American Journal of Respiratory Cell and Molecular Biology}, author={Park, J. and Fang, S. J. and Crews, A. L. and Lin, K. W. and Adler, K. B.}, year={2008}, pages={68–76} }
 @article{kraft_adler_ingram_crews_atkinson_cairns_krause_chu_2008, title={Mycoplasma pneumoniae increases airway epithelial cell MUC5AC expression in asthma.}, volume={34}, journal={European Respiratory Journal}, author={Kraft, M. and Adler, K. B. and Ingram, J. L. and Crews, A. L. and Atkinson, T. P. and Cairns, C. B. and Krause, D. C. and Chu, H. W.}, year={2008}, pages={43–46} }
 @article{kraft_adler_ingram_crews_atkinson_cairns_krause_chu_2008, title={Mycoplasma pneumoniae induces airway epithelial cell expression of MUC5AC in asthma}, volume={31}, ISSN={["1399-3003"]}, DOI={10.1183/09031936.00103307}, abstractNote={As excess mucin expression can contribute to the exacerbation of asthma, the present authors hypothesised that Mycoplasma pneumoniae significantly induces MUC5AC (the major airway mucin) expression in airway epithelial cells isolated directly from asthmatic subjects. A total of 11 subjects with asthma and six normal controls underwent bronchoscopy with airway brushing. Epithelial cells were cultured at an air–liquid interface and incubated with and without M. pneumoniae for 48 h, and in the presence and absence of nuclear factor (NF)-κB and a toll-like receptor (TLR)2 inhibitor. Quantitative PCR was performed for MUC5AC and TLR2 mRNA. MUC5AC protein and total protein were determined by ELISA. M. pneumoniae exposure significantly increased MUC5AC mRNA and protein expression after 48 h in epithelial cells isolated from asthmatic, but not from normal control subjects, at all concentrations as compared to unexposed cells. TLR2 mRNA expression was significantly increased in asthmatic epithelial cells at 4 h compared with unexposed cells. NF-κB and TLR2 inhibition reduced MUC5AC expression to the level of the unexposed control in both groups. Mycoplasma pneumoniae exposure significantly increased MUC5AC mRNA and protein expression preferentially in airway epithelial cells isolated from asthmatic subjects. The toll-like receptor 2 pathway may be involved in this process.}, number={1}, journal={EUROPEAN RESPIRATORY JOURNAL}, author={Kraft, M. and Adler, K. B. and Ingram, J. L. and Crews, A. L. and Atkinson, T. P. and Cairns, C. B. and Krause, D. C. and Chu, H. W.}, year={2008}, month={Jan}, pages={43–46} }
 @misc{waterman_adler_2008, title={Oxidant stress and airway epithelial function}, volume={61}, journal={Free radical effects on membranes}, author={Waterman, J. T. and Adler, K. B.}, year={2008}, pages={243–255} }
 @misc{lin_park_crews_li_adler_2008, title={Protease-activated receptor-2 (PAR-2) is a weak enhancer of mucin secretion by human bronchial epithelial cells in vitro}, volume={40}, ISSN={["1878-5875"]}, DOI={10.1016/j.biocel.2007.10.031}, abstractNote={PAR-2, a member of a family of G-protein-coupled receptors, can be activated by serine proteases via proteolytic cleavage. PAR-2 expression is known to be upregulated in respiratory epithelium subsequent to inflammation in asthma and chronic obstructive pulmonary disease (COPD). Since these diseases also are characterized by excessive mucus production and secretion, we investigated whether PAR-2 could be linked to mucin hypersecretion by airway epithelium. Normal human bronchial epithelial (NHBE) cells in primary culture or the human bronchial epithelial cell lines, NCI-H292 and HBE-1, were used. NHBE, NCI-H292, and HBE-1 cells expressed prominent levels of PAR-2 protein. Short-term (30min) exposure of cells to the synthetic PAR-2 agonist peptide (SLIGKV-NH2) elicited a small but statistically significant increase in mucin secretion at high concentrations (100microM and 1000microM), compared to a control peptide with reversed amino acid sequence (VKGILS-NH2). Neither human lung tryptase nor bovine pancreatic trypsin, both PAR-2 agonists, affected NHBE cell mucin secretion when added over a range of concentrations. Knockdown of PAR-2 expression by siRNA blocked the stimulatory effect of the AP. The results suggest that, since PAR-2 activation only weakly increases mucin secretion by human airway epithelial cells in vitro, PAR-2 probably is not a significant contributor to mucin hypersecretion in inflamed airways.}, number={6-7}, journal={INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY}, author={Lin, Ko-Wei and Park, Joungjoa and Crews, Anne L. and Li, Yuehua and Adler, Kenneth B.}, year={2008}, pages={1379–1388} }
 @misc{wang_bai_li_adler_wang_2008, title={Role of airway epithelial cells in development of asthma and allergic rhinitis}, volume={102}, ISSN={["0954-6111"]}, DOI={10.1016/j.rmed.2008.01.017}, abstractNote={<h2>Summary</h2> Asthma and allergic rhinitis frequently coexist in the same patient. There is a similarity and variation as well as potential relationship between asthma and allergic rhinitis. There is an increasing evidence to suggest a major involvement of airway epithelial cells in the pathogenesis of asthma and allergic rhinitis. The present review describes the importance of the airway epithelial cell in the development of allergic airway diseases, its role as the primary airway defense against exposure of the airway and lung to inflammatory stimuli and antigens and as an important player through activation of epithelial Toll-like receptors (TLRs) to provide an important link between innate immunity and allergic disease. Additionally, airway epithelial cells can act as inflammatory promoters capable of directing dendritic cells (DCs) towards a T helper 2 (Th2) response, and as active producers of several inflammatory/anti-inflammatory mediators. It is hypothesized that airway epithelial cells may play as both inflammatory initiator and immuno-pathological feedback regulation between allergic rhinitis and asthma via release of systemic inflammatory mediators. Thus, airway epithelial cells may be valuable therapeutic targets for discovery and development of new drugs and/or new therapeutic strategies to treat asthma and allergic rhinitis.}, number={7}, journal={RESPIRATORY MEDICINE}, author={Wang, YaoLi and Bai, Chunxue and Li, Ka and Adler, Kenneth B. and Wang, Xiangdong}, year={2008}, month={Jul}, pages={949–955} }
 @article{green_eckert_sharief_crews_adler_jones_2007, title={A peptide against the N-terminus of MARCKS protein attenuates leukocyte migration.}, volume={175}, journal={404nOtfound}, author={Green, T. D. and Eckert, B. S. and Sharief, Y. and Crews, A. L. and Adler, K. B. and Jones, S. L.}, year={2007}, pages={A915} }
 @article{park_takashi_crews_miller-larsson_adler_2007, title={Anti-inflammatory effects of budesonide and formoterol in primary cultures of human airway epithelial cells.}, volume={175}, journal={404nOtfound}, author={Park, J. J. and Takashi, S. and Crews, A. L. and Miller-Larsson, A. and Adler, K. B.}, year={2007}, pages={A178} }
 @article{agrawal_rengarajan_adler_ram_ghosh_fahim_dickey_2007, title={Inhibition of mucin secretion with MARCKS-related peptide improves airway obstruction in a mouse model of asthma}, volume={102}, ISSN={["1522-1601"]}, DOI={10.1152/japplphysiol.00630.2006}, abstractNote={ Allergic asthma is associated with airway epithelial cell mucous metaplasia and mucin hypersecretion, but the consequences of mucin hypersecretion on airway function are unclear. Recently, a peptide derived from the myristoylated alanine-rich C kinase substrate protein NH2-terminal sequence (MANS) was shown to inhibit methacholine (MCh)-induced mucin secretion from airway mucous cells by >90%. We studied the effect of intranasal pretreatment with this peptide on specific airway conductance (sGaw) during challenge with MCh in mice with allergen-induced mucous cell metaplasia. sGaw was noninvasively measured in spontaneously breathing restrained mice, using a double-chamber plethysmograph. Pretreatment with MANS peptide, but not a control peptide [random NH2-terminal sequence (RNS)], resulted in partial inhibition of the fall in sGaw induced by 60 mM MCh (mean ± SE; baseline 1.15 ± 0.06; MANS/MCh 0.82 ± 0.05; RNS/MCh 0.55 ± 0.05 cmH2O/s). The protective effect of MANS was also seen in mice challenged with allergen for 3 consecutive days to increase airway hyperresponsiveness, although the degree of protection was less (baseline 1.1 ± 0.08; MANS/MCh, 0.65 ± 0.06; RNS/MCh 0.47 ± 0.03 cmH2O/s). Because routine sGaw measurement in mice includes nasal airways, the effectiveness of MANS was also confirmed in mice breathing through their mouths after nasal occlusion (baseline 0.92 ± 0.05; MANS/MCh 0.83 ± 0.06; RNS/MCh 0.61 ± 0.03 cmH2O/s). In all instances, sGaw in the MANS-pretreated group was ∼35% higher than in RNS-treated controls, and mucous obstruction accounted for ∼50% of the MCh-induced fall in sGaw. In summary, mucin secretion has a significant role in airway obstruction in a mouse model of allergic asthma, and strategies to inhibit mucin secretion merit further investigation. }, number={1}, journal={JOURNAL OF APPLIED PHYSIOLOGY}, author={Agrawal, A. and Rengarajan, S. and Adler, K. B. and Ram, A. and Ghosh, B. and Fahim, M. and Dickey, B. F.}, year={2007}, month={Jan}, pages={399–405} }
 @misc{li_martin_adler_2007, title={Method and compositions for altering mucus secretion}, volume={7,265,088}, number={2007 Sept. 4}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Li, Y.-H. and Martin, L. D. and Adler, K. B.}, year={2007} }
 @article{kraft_nijira lugogo_adler_crews_moss_stalls_church_slade_beaver_chu_2007, title={Mycoplasma pneumoniae increases airway epithelial cell MUC5AC expression in asthma.}, volume={175}, journal={404nOtfound}, author={Kraft, M. and Nijira Lugogo, H. W. and Adler, K. B. and Crews, A. L. and Moss, T. A. and Stalls, M. A. and Church, T. D. and Slade, D. J. and Beaver, D. M. and Chu, H. W.}, year={2007}, pages={A324} }
 @article{park_adler_2007, title={Potential therapy for mucus hypersecretion in COPD.}, volume={3}, DOI={10.1080/17471060601063231}, abstractNote={Mucus hypersecretion is a major pathophysiologic feature of chronic bronchitis. Although mucus functions as a barrier and a facilitator of mucociliary clearance, persistent mucus hypersecretion results in airway obstruction and compromised clearance of inhaled bacteria and particles from the airways of patients with chronic obstructive pulmonary disease. Treatment of mucus hypersecretion is a major therapeutic target; however, mechanisms of mucus hypersecretion remain unknown. Herein, we present evidence that human neutrophil elastase (HNE), a pathophysiologically relevant stimulant of mucus hypersecretion in the airways of patients with chronic bronchitis, provokes release of mucin (the glycoprotein component of mucus) by human airway epithelial cells in vitro. Signaling molecules involved in HNE-induced mucin hypersecretion include protein kinase C, specifically the delta isoform, and the myristoylated alanine-rich C kinase substrate protein. These molecules represent potential therapeutic targets for regulating mucin secretion in patients.}, journal={404nOtfound}, author={Park, J.-A. and Adler, K. B.}, year={2007}, pages={66–71} }
 @article{park_crews_lampe_fang_park_adler_2007, title={Protein kinase C delta regulates airway mucin secretion via phosphorylation of MARCKS protein}, volume={171}, ISSN={["1525-2191"]}, DOI={10.2353/ajpath.2007.070318}, abstractNote={Mucin hypersecretion is a major pathological feature of many respiratory diseases, yet cellular mechanisms regulating secretion of mucin have not been fully elucidated. Previously, we reported that mucin hypersecretion induced by human neutrophil elastase involves activation of protein kinase C (PKC), specifically the δ-isoform (PKCδ). Here, we further investigated the role of PKCδ in mucin hypersecretion using both primary human bronchial epithelial cells and the human bronchial epithelial 1 cell line as in vitro model systems. Phorbol-12-myristate-13-acetate (PMA)-induced mucin hypersecretion was significantly attenuated by rottlerin, a PKCδ-selective inhibitor. Rottlerin also reduced PMA- or human neutrophil elastase-induced phosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS) protein in these cells. Both secretion and MARCKS phosphorylation were significantly enhanced by the PKCδ activator bryostatin 1. A dominant-negative PKCδ construct (pEGFP-N1/PKCδK376R) transfected into human bronchial epithelial 1 cells significantly attenuated both PMA-induced mucin secretion and phosphorylation of MARCKS, whereas transfection of a wild-type construct increased PKCδ and enhanced mucin secretion and MARCKS phosphorylation. Similar transfections of a dominant-negative or wild-type PKCε construct did not affect either mucin secretion or MARCKS phosphorylation. The results suggest that PKCδ plays an important role in mucin secretion by airway epithelium via regulation of MARCKS phosphorylation. Mucin hypersecretion is a major pathological feature of many respiratory diseases, yet cellular mechanisms regulating secretion of mucin have not been fully elucidated. Previously, we reported that mucin hypersecretion induced by human neutrophil elastase involves activation of protein kinase C (PKC), specifically the δ-isoform (PKCδ). Here, we further investigated the role of PKCδ in mucin hypersecretion using both primary human bronchial epithelial cells and the human bronchial epithelial 1 cell line as in vitro model systems. Phorbol-12-myristate-13-acetate (PMA)-induced mucin hypersecretion was significantly attenuated by rottlerin, a PKCδ-selective inhibitor. Rottlerin also reduced PMA- or human neutrophil elastase-induced phosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS) protein in these cells. Both secretion and MARCKS phosphorylation were significantly enhanced by the PKCδ activator bryostatin 1. A dominant-negative PKCδ construct (pEGFP-N1/PKCδK376R) transfected into human bronchial epithelial 1 cells significantly attenuated both PMA-induced mucin secretion and phosphorylation of MARCKS, whereas transfection of a wild-type construct increased PKCδ and enhanced mucin secretion and MARCKS phosphorylation. Similar transfections of a dominant-negative or wild-type PKCε construct did not affect either mucin secretion or MARCKS phosphorylation. The results suggest that PKCδ plays an important role in mucin secretion by airway epithelium via regulation of MARCKS phosphorylation. Mucus produced by epithelium of respiratory, gastrointestinal, and reproductive tracts provides a barrier between the external environment and cellular components of the epithelial layer. Mucins, the glycoprotein component of mucus, constitute a family of large, highly glycosylated macromolecules that impart physical (aggregation, viscosity, viscoelasticity, and lubrication) and biological (protection) properties to mucus (reviewed in Ref. 1Rose MC Mucins: structure, function, and role in pulmonary diseases.Am J Physiol. 1992; 263: L413-L429PubMed Google Scholar). Airway mucus is an integral component of the mucociliary clearance system in the trachea and bronchi and thus serves to protect the lower airways and alveoli from impingement of particulate matter and pathogens. However, mucin secretion is abnormally augmented in disease states, such as chronic bronchitis, asthma, and cystic fibrosis, increasing morbidity and mortality in these patients (reviewed in Refs. 1Rose MC Mucins: structure, function, and role in pulmonary diseases.Am J Physiol. 1992; 263: L413-L429PubMed Google Scholar and 2Rogers DF Barnes PJ Treatment of airway mucus hypersecretion.Ann Med. 2006; 38: 116-125Crossref PubMed Scopus (172) Google Scholar). Mucin hypersecretion is potentiated by many pathophysiological mediators, such as bacterial proteinases and endotoxin, adenine and guanine nucleotides, cytokines, inflammatory mediators, and eicosanoids (reviewed in Ref. 3Adler KB Li Y Airway epithelium and mucus: intracellular signaling pathways for gene expression and secretion.Am J Respir Cell Mol Biol. 2001; 25: 397-400Crossref PubMed Scopus (50) Google Scholar). Intracellular mechanisms and signaling molecules involved in the secretory process have not been fully elucidated.Protein kinase C (PKC) is a serine/threonine kinase involved in various exocytotic events in different cell types, including secretion of mucin,4Abdullah LH Bundy JT Ehre C Davis CW Mucin secretion and PKC isoforms in SPOC1 goblet cells: differential activation by purinergic agonist and PMA.Am J Physiol Lung Cell Mol Physiol. 2003; 285: L149-L160PubMed Google Scholar, 5Plaisancie P Ducroc R El Homsi M Tsocas A Guilmeau S Zoghbi S Thibaudeau O Bado A Luminal leptin activates mucin-secreting goblet cells in the large bowel.Am J Physiol Gastrointest Liver Physiol. 2006; 290: G805-G812Crossref PubMed Scopus (59) Google Scholar insulin,6Yaney GC Fairbanks JM Deeney JT Korchak HM Tornheim K Corkey BE Potentiation of insulin secretion by phorbol esters is mediated by PKC-alpha and nPKC isoforms.Am J Physiol Endocrinol Metab. 2002; 283: E880-E888Crossref PubMed Scopus (34) Google Scholar neurotransmitters,7Shoji-Kasai Y Itakura M Kataoka M Yamamori S Takahashi M Protein kinase C-mediated translocation of secretory vesicles to plasma membrane and enhancement of neurotransmitter release from PC12 cells.Eur J Neurosci. 2002; 15: 1390-1394Crossref PubMed Scopus (50) Google Scholar and platelet dense granules.8Murugappan S Tuluc F Dorsam RT Shankar H Kunapuli SP Differential role of protein kinase C delta isoform in agonist-induced dense granule secretion in human platelets.J Biol Chem. 2004; 279: 2360-2367Crossref PubMed Scopus (92) Google Scholar Previously, we demonstrated that mucin secretion in airway epithelial cells is regulated by PKC via phosphorylation of the myristoylated alanine-rich C kinase substrate (MARCKS).9Li Y Martin LD Spizz G Adler KB MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro.J Biol Chem. 2001; 276: 40982-40990Crossref PubMed Scopus (150) Google Scholar, 10Singer M Martin LD Vargaftig BB Park J Gruber AD Li Y Adler KB A MARCKS-related peptide blocks mucus hypersecretion in a mouse model of asthma.Nat Med. 2004; 10: 193-196Crossref PubMed Scopus (147) Google Scholar In addition, we demonstrated that mucin hypersecretion in human airway epithelial cells in vitro in response to human neutrophil elastase (HNE) appears to be mediated by the δ-isoform of PKC (PKCδ).11Park JA He F Martin LD Li Y Chorley BN Adler KB Human neutrophil elastase induces hypersecretion of mucin from well-differentiated human bronchial epithelial cells in vitro via a protein kinase C{delta}-mediated mechanism.Am J Pathol. 2005; 167: 651-661Abstract Full Text Full Text PDF PubMed Scopus (96) Google Scholar Not surprisingly, PKCδ, a novel PKC isoform, has a strong affinity for MARCKS and can phosphorylate MARCKS both in vitro and in vivo.12Fujise A Mizuno K Ueda Y Osada S Hirai S Takayanagi A Shimizu N Owada MK Nakajima H Ohno S Specificity of the high affinity interaction of protein kinase C with a physiological substrate, myristoylated alanine-rich protein kinase C substrate.J Biol Chem. 1994; 269: 31642-31648Abstract Full Text PDF PubMed Google Scholar, 13Herget T Oehrlein SA Pappin DJ Rozengurt E Parker PJ The myristoylated alanine-rich C-kinase substrate (MARCKS) is sequentially phosphorylated by conventional, novel and atypical isotypes of protein kinase C.Eur J Biochem. 1995; 233: 448-457Crossref PubMed Scopus (79) Google Scholar, 14Cabell CH Verghese GM Rankl NB Burns DJ Blackshear PJ MARCKS phosphorylation by individual protein kinase C isozymes in insect Sf9 cells.Proc Assoc Am Physicians. 1996; 108: 37-46PubMed Google Scholar Increasing evidence suggests that PKCδ mediates exocytotic secretion in several different cell types.4Abdullah LH Bundy JT Ehre C Davis CW Mucin secretion and PKC isoforms in SPOC1 goblet cells: differential activation by purinergic agonist and PMA.Am J Physiol Lung Cell Mol Physiol. 2003; 285: L149-L160PubMed Google Scholar, 8Murugappan S Tuluc F Dorsam RT Shankar H Kunapuli SP Differential role of protein kinase C delta isoform in agonist-induced dense granule secretion in human platelets.J Biol Chem. 2004; 279: 2360-2367Crossref PubMed Scopus (92) Google Scholar, 15Ishikawa T Iwasaki E Kanatani K Sugino F Kaneko Y Obara K Nakayama K Involvement of novel protein kinase C isoforms in carbachol-stimulated insulin secretion from rat pancreatic islets.Life Sci. 2005; 77: 462-469Crossref PubMed Scopus (12) Google Scholar, 16Leitges M Gimborn K Elis W Kalesnikoff J Hughes MR Krystal G Huber M Protein kinase C-delta is a negative regulator of antigen-induced mast cell degranulation.Mol Cell Biol. 2002; 22: 3970-3980Crossref PubMed Scopus (115) Google Scholar, 17Cho SH Woo CH Yoon SB Kim JH Protein kinase Cdelta functions downstream of Ca2+ mobilization in FcepsilonRI signaling to degranulation in mast cells.J Allergy Clin Immunol. 2004; 114: 1085-1092Abstract Full Text Full Text PDF PubMed Scopus (55) Google ScholarHere, we further elucidate the role of PKCδ in the mucin secretory pathway in human airway epithelial cells in vitro. The mucin secretory response and phosphorylation of MARCKS were assessed after exposure of well differentiated normal human bronchial epithelial (NHBE) cells to phorbol-12-myristate-13-acetate (PMA), a general PKC activator, or bryostatin 1, a PKCδ/ε activator. In addition, we used the papilloma virus-transformed human bronchial epithelial 1 (HBE1) cell line for molecular manipulations. A dominant-negative PKCδ construct (K376R) transfected into HBE1 cells attenuated PMA-stimulated mucin secretion and MARCKS phosphorylation, whereas a similar dominant-negative PKCε construct was without effect. The results indicate that PKCδ is a key isoform regulating airway mucin secretion, and the mechanism of its action appears to involve phosphorylation of MARCKS protein.Materials and MethodsCulture of Bronchial Epithelial CellsPrimary NHBE cells purchased from Cambrex Bioscience (Walkersville, MD) were expanded and maintained in a humidified air/5% CO2 incubator as described previously.9Li Y Martin LD Spizz G Adler KB MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro.J Biol Chem. 2001; 276: 40982-40990Crossref PubMed Scopus (150) Google Scholar, 11Park JA He F Martin LD Li Y Chorley BN Adler KB Human neutrophil elastase induces hypersecretion of mucin from well-differentiated human bronchial epithelial cells in vitro via a protein kinase C{delta}-mediated mechanism.Am J Pathol. 2005; 167: 651-661Abstract Full Text Full Text PDF PubMed Scopus (96) Google Scholar Cells from at least three separate donors were used in these studies. Passage 31 or 32 of human papilloma virus-transformed human bronchial epithelial cells (HBE1)18Yankaskas JR Haizlip JE Conrad M Koval D Lazarowski E Paradiso AM Rinehart Jr, CA Sarkadi B Schlegel R Boucher RC Papilloma virus immortalized tracheal epithelial cells retain a well-differentiated phenotype.Am J Physiol. 1993; 264: C1219-C1230PubMed Google Scholar was seeded and maintained as described previously. Transient transfection of HBE1 cells was performed after 10 days of culture in air-liquid interface.Exposure of Cells to Inhibitors or SecretagoguesWell differentiated NHBE cells were exposed to test agents both apically and basolaterally for 15 minutes (unless otherwise indicated). Transfected HBE1 cells were exposed to PMA (EMD Biosciences, La Jolla, CA) applied apically only. Cells were preincubated with the PKCδ-selective inhibitor, rottlerin (EMD Biosciences) for 20 minutes before PMA exposure. Initial stock solutions of rottlerin or PMA were prepared in dimethyl sulfoxide, kept at −20°C, and diluted in growth medium directly before use. When cells were exposed to PMA in the presence or absence of rottlerin, PMA was “spiked” into each well at the indicated concentration.Measurement of Mucin SecretionMucin was collected both at baseline and after treatments as described previously.9Li Y Martin LD Spizz G Adler KB MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro.J Biol Chem. 2001; 276: 40982-40990Crossref PubMed Scopus (150) Google Scholar Baseline mucin secretion was used to normalize well-to-well variation. After baseline mucin samples were collected, cells were rested overnight and exposed to test reagents the next day for indicated periods of time. After each treatment period, secreted mucin was collected as the baseline sample and quantified by sandwich enzyme-linked immunosorbent assay using the 17Q2 antibody (Covance Research Products, Berkeley, CA), a monoclonal antibody that reacts specifically with a carbohydrate epitope on human airway mucins.19Lin H Carlson DM St George JA Plopper CG Wu R An ELISA method for the quantitation of tracheal mucins from human and nonhuman primates.Am J Respir Cell Mol Biol. 1989; 1: 41-48Crossref PubMed Scopus (63) Google Scholar The 17Q2 antibody was purified using an ImmunoPure(G) IgG purification kit (Pierce Biotechnology, Rockford, IL) following the manufacturer's protocol and then conjugated with alkaline phosphatase (EMD Biosciences). To account for variability between cultures and experiments, levels of mucin secretion were reported as percentage of the nontreated control. Actual values for mucin released by control cell cultures in these experiments ranged from 40 to 70 ng/ml in NHBE cells and 7 to 10 ng/ml in HBE1 cells.Subcellular Localization of PKC IsoformsActivation of PKCδ was assessed by subcellular fractionation following the protocol described by Kajstura et al20Kajstura J Cigola E Malhotra A Li P Cheng W Meggs LG Anversa P Angiotensin II induces apoptosis of adult ventricular myocytes in vitro.J Mol Cell Cardiol. 1997; 29: 859-870Abstract Full Text PDF PubMed Scopus (375) Google Scholar and subsequent Western blot analysis using a PKCδ-specific antibody (Cell Signaling Technology, Inc., Danvers, MA). Briefly, cells were washed with cold PBS and scraped into lysis buffer [20 mmol/L Tris-Cl (pH 7.5), 1 mmol/L ethylenediamine tetraacetic acid, 100 mmol/L NaCl, 1 mmol/L phenylmethylsulfonyl fluoride, 1 mmol/L dithiothreitol, 1% (v/v) protease inhibitor cocktail, and phosphatase inhibitor cocktail (Sigma, St. Louis, MO)]. The lysate was then sonicated and pelleted at 20,000 × g (Eppendorf 5417 centrifuge) for 40 minutes. The supernatant was collected and kept as the cytosolic fraction at −80°C until used. The remaining pellet was resuspended in lysis buffer containing 1% Triton X-100, sonicated, and centrifuged at 20,000 × g for 40 minutes. The supernatant membrane fraction was stored at −80°C until analyzed by Western blot.Western Blot AnalysisTotal MARCKS, phosphorylated MARCKS, PKCδ, and PKCε protein levels were measured via Western blot. The protein concentrations of cell lysates were quantified by a Bradford assay (Bio-Rad Laboratories, Hercules, CA). Sample lysates were prepared by boiling in 2× SDS sample buffer [125 mmol/L Tris-Cl (pH 6.8), 25% glycerol, 4% SDS, 10% β-mercaptoethanol, and 0.04% bromphenol blue] for 10 minutes. Sample lysates (30 to 60 μg) were loaded on 10 or 12% SDS-polyacrylamide gels and then transferred to a polyvinylidene difluoride membrane (Schleicher & Schuell BioScience, Inc., Keene, NH) following electrophoresis. Polyvinylidene difluoride membranes were blocked with 5% nonfat milk and then probed with an appropriate dilution of primary antibody followed by horseradish peroxidase-conjugated anti-mouse or anti-rabbit antibodies. Chemiluminescent detection was performed using ECL detection reagents (GE Health care Life Sciences, Piscataway, NJ) following the manufacturer's protocol. Amounts of specific proteins in bands were quantified using Labworks image acquisition and analysis software 4.0. (Ultra Violet Products, Ltd., Upland, CA).Antibodies against α-tubulin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) and E-cadherin (BD Biosciences, San Jose, CA) were used as loading controls for cytosolic and membrane fractions, respectively. Phosphorylated MARCKS (at serine 152/156) was detected with a specific antibody (Cell Signaling Technology, Inc.). After detection, the membrane was stripped in 62.5 mmol/L Tris-Cl (pH 6.5), 10% SDS, and 100 mmol/L β-mercaptoethanol for 10 minutes at room temperature and reprobed with a monoclonal antibody against total MARCKS protein (clone no. 2F12; Upstate, Charlottesville, VA) to verify equal loading.Transient Transfection of PKC ConstructsTransient transfection of vectors overexpressing wild-type or dominant-negative PKCδ and PKCε in HBE1 cells was performed using the FuGene 6 transfection reagent (Roche Applied Science, Indianapolis, IN) following the manufacturer's protocol. The pEGFP-N1 vectors containing a wild-type PKCδ cDNA21Mischak H Pierce JH Goodnight J Kazanietz MG Blumberg PM Mushinski JF Phorbol ester-induced myeloid differentiation is mediated by protein kinase C-alpha and -delta and not by protein kinase C-beta II, -epsilon, -zeta, and -eta.J Biol Chem. 1993; 268: 20110-20115Abstract Full Text PDF PubMed Google Scholar and a dominant-negative PKCδ mutant cDNA [lysine (AAG)→arginine (AGG) mutation, position 376]22Li L Lorenzo PS Bogi K Blumberg PM Yuspa SH Protein kinase Cdelta targets mitochondria, alters mitochondrial membrane potential, and induces apoptosis in normal and neoplastic keratinocytes when overexpressed by an adenoviral vector.Mol Cell Biol. 1999; 19: 8547-8558Crossref PubMed Google Scholar were generously provided by Dr. Arti Shukla (University of Vermont, Burlington, VT) and Dr. Peter Blumberg (National Cancer Institute, Bethesda, MD). The K376→R mutation in the ATP binding site of the catalytic domain has been demonstrated previously to inhibit PKCδ kinase activity.23Li W Yu JC Shin DY Pierce JH Characterization of a protein kinase C-delta (PKC-delta) ATP binding mutant: an inactive enzyme that competitively inhibits wild type PKC-delta enzymatic activity.J Biol Chem. 1995; 270: 8311-8318Crossref PubMed Scopus (89) Google Scholar Briefly, HBE1 cells grown in air/liquid interface were dissociated in versene solution (Invitrogen, Carlsbad, CA) and re-seeded in 12-well culture plates at a density of 1 × 105 cells/cm2. After overnight incubation, cells were transfected with the pEGFP-N1 vector alone or the pEGFP-N1 vector containing either a wild-type or dominant-negative PKCδ cDNA (K376R). Isotype controls for the dominant-negative PKCδ consisted of transient transfection of both wild-type PKCε as well as a catalytically inactive dominant-negative PKCε (K437R) construct-tagged with hemagglutin24Lee YJ Soh JW Jeoung DI Cho CK Jhon GJ Lee SJ Lee YS PKC epsilon -mediated ERK1/2 activation involved in radiation-induced cell death in NIH3T3 cells.Biochim Biophys Acta. 2003; 1593: 219-229Crossref PubMed Scopus (32) Google Scholar (PKCε constructs were kindly provided by Dr. Jae-Won Soh, University of Inha, Inha, Republic of Korea). Cells were subsequently cultured for 48 hours to allow for detectable protein expression. Transfection of PKCδ constructs was confirmed by fluorescent microscopy and assessment of expression of green fluorescence protein (GFP)-tagged PKCδ assessed by Western blot analysis using monoclonal antibodies against PKCδ or GFP (Cell Signaling Technology, Inc.). Transfection of PKCε was confirmed by Western blot analysis using PKCε and HA monoclonal antibodies (Covance Research Products).Cytotoxicity AssayAll treatments used were tested for cytotoxicity using a CytoTox 96 nonradioactive cytotoxicity assay kit (Promega, Madison, WI) according to the manufacturer's instructions. The results were expressed as the ratio of released lactate dehydrogenase to total lactate dehydrogenase. Released lactate dehydrogenase never exceeded 10% of total lactate dehydrogenase with any of the treatments (data not shown).Statistical AnalysisData were expressed as the ratio of treatment to the corresponding vehicle (dimethyl sulfoxide or media) control. Results were evaluated using one-way analysis of variance with Dunnett's test and a Bonferroni posttest correction for multiple comparisons.25Kleinbaum DG Kupper LL Muller KE Applied Regression Analysis and Other Multivariable Methods. PWS-Kent Publishing Co., Boston, MA1988Google Scholar A P value of less than 0.05 was considered significant.ResultsEffect of Rottlerin on PMA-Induced Mucin Secretion and MARCKS Phosphorylation in NHBE CellsTo determine whether PKCδ is an important regulatory molecule in mucin secretion, we investigated the effect of rottlerin, a PKCδ-selective inhibitor, on PMA-induced mucin secretion in well differentiated NHBE cells. As illustrated in Figure 1A, 100 nmol/L PMA provoked translocation of PKCδ from cytosol to membrane in these cells. Inhibition of PKCδ activity by pretreatment of cells with rottlerin (1 to 10 μmol/L) for 20 minutes significantly attenuated PMA-induced mucin secretion in a concentration-dependent manner (Figure 1B). Phosphorylation of MARCKS mediated by PMA in the presence or absence of 15 μmol/L rottlerin was analyzed by Western blot. HNE, previously shown to stimulate mucin secretion in NHBE cells via a rottlerin-inhibitable mechanism,11Park JA He F Martin LD Li Y Chorley BN Adler KB Human neutrophil elastase induces hypersecretion of mucin from well-differentiated human bronchial epithelial cells in vitro via a protein kinase C{delta}-mediated mechanism.Am J Pathol. 2005; 167: 651-661Abstract Full Text Full Text PDF PubMed Scopus (96) Google Scholar was used as an additional control. As shown in Figure 1C, both PMA- and HNE-induced phosphorylation of MARCKS were decreased by pretreatment with rottlerin.Bryostatin 1 Provokes Mucin Secretion and MARCKS Phosphorylation in NHBE CellsBryostatin 1, a PKCδ/ε activator, was used to investigate further the relationship between PKCδ and mucin secretion in NHBE cells. A naturally occurring marine invertebrate-derived cyclic lactone, Bryostatin 1, causes rapid activation of PKCδ and prolonged protection of PKCδ against ubiquitination and proteolysis.26Szallasi Z Denning MF Smith CB Dlugosz AA Yuspa SH Pettit GR Blumberg PM Bryostatin 1 protects protein kinase C-delta from down-regulation in mouse keratinocytes in parallel with its inhibition of phorbol ester-induced differentiation.Mol Pharmacol. 1994; 46: 840-850PubMed Google Scholar, 27Heit I Wieser RJ Herget T Faust D Borchert-Stuhltrager M Oesch F Dietrich C Involvement of protein kinase Cdelta in contact-dependent inhibition of growth in human and murine fibroblasts.Oncogene. 2001; 20: 5143-5154Crossref PubMed Scopus (33) Google Scholar Bryostatin 1 interacts with the diacylglycerol binding site on PKC,28Mutter R Wills M Chemistry and clinical biology of the bryostatins.Bioorg Med Chem. 2000; 8: 1841-1860Crossref PubMed Scopus (179) Google Scholar but its complete mode of action has not been fully elucidated, and it can affect PKCs in a cell-type specific manner.29Szallasi Z Smith CB Pettit GR Blumberg PM Differential regulation of protein kinase C isozymes by bryostatin 1 and phorbol 12-myristate 13-acetate in NIH 3T3 fibroblasts.J Biol Chem. 1994; 269: 2118-2124Abstract Full Text PDF PubMed Google Scholar As illustrated in Figure 2, exposure of NHBE cells to bryostatin 1 over a range of concentrations (10 to 1000 nmol/L) for 15 minutes resulted in translocation of PKCδ from cytosol to membrane in response to all concentrations tested (Figure 2A). Mucin secretion was also significantly increased by bryostatin 1, with maximal stimulation at 100 nmol/L (Figure 2B). As illustrated in Figure 2C, phosphorylation of MARCKS also was induced in these cells in response to bryostatin 1 (from 10 to 1000 nmol/L) with maximal phosphorylation at 100 nmol/L. None of these treatments induced cytotoxicity as measured by lactate dehydrogenase release assay (data not shown).Figure 2Effect of bryostatin 1, a PKCδ activator, on mucin secretion in well differentiated NHBE cells. NHBE cells were exposed to bryostatin 1 over a range of concentrations from 1 to 1000 nmol/L for 15 minutes. A: PKCδ translocates from cytosol to membrane in response to bryostatin 1. α-Tubulin and E-cadherin were used as controls for the cytosolic and membrane fractions, respectively. Blots are representative of three replicate experiments. B: Bryostatin 1 provokes mucin secretion by NHBE cells in a concentration-dependent manner. Significantly different from vehicle control: *P < 0.05; †P < 0.001; ‡P < 0.005. Data are presented as mean ± SEM (n = 4). C: Phosphorylation of MARCKS in NHBE cells is induced by bryostatin 1 in a concentration-dependent manner. Blots are representative of three replicate experiments.View Large Image Figure ViewerDownload Hi-res image Download (PPT)PKC Activation Stimulates Mucin Secretion in HBE1 CellsTo investigate further a role for PKCδ as a regulator of mucin secretion using molecular manipulation of PKCδ activity, the HBE1 cell line was used. As illustrated in Figure 3, exposure of HBE1 cells to 500 nmol/L PMA for 15 minutes significantly increased mucin secretion (by ∼1.7-fold compared with medium vehicle control) and also induced phosphorylation of MARCKS in these cells.Figure 3HBE-1 cells secrete mucin in response to PKC activation. HBE-1 cells maintained in air/liquid interface were exposed to 100 or 500 nmol/L PMA for 15 minutes. Mucin secretion and phosphorylation of MARCKS were assessed by enzyme-linked immunosorbent assay and Western blot analysis, respectively. A: Mucin secretion is significantly enhanced by PMA at 500 nmol/L (but not 100 nmol/L) in HBE1 cells. *Significantly different from vehicle control (P < 0.05). Data are presented as mean ± SEM (n = 4). B: Phosphorylation of MARCKS is increased by exposure of HBE1 cells to 500 nmol/L PMA. Blots are representative of three replicate experiments.View Large Image Figure ViewerDownload Hi-res image Download (PPT)PKCδ Appears to Regulate Mucin Secretion in Airway Epithelial CellsTransient transfection of the PKCδ and PKCε constructs into HBE1 cells was confirmed by fluorescent microscopy and Western blot analysis. After 48 hours of transfection, GFP expressed in the transfected cells was detected with a fluorescent microscope [Eclipse TE300 (Nikon, Tokyo, Japan) or Axiovert 35 (Zeiss, Welwyn Garden City, UK)] before PMA exposure (data not shown). After exposure to PMA, cells were lysed to detect expression of PKCδ protein fused with GFP via Western blot analysis using anti-PKCδ and -GFP antibodies (data not shown). Transfection of PKCε was confirmed by Western blot analysis using an antibody against PKCε and an HA tag. Transfection efficiency of all constructs and controls was about 20 to 25% as determined by quantification of GFP and X-gal assay for PKCδ and PKCε, respectively (data not shown).As illustrated in Figure 4A, mucin secretion by transfected HBE1 cells was stimulated by exposure to 500 nmol/L PMA. Transfection of HBE1 cells with the dominant-negative PKCδ construct (pEGFP-N1/PKCδK376R) resulted in significant reduction of PMA-induced mucin secretion (∼45%), whereas cells transfected with the wild-type PKCδ construct (pEGFP-N1/PKCδ) showed a significant enhancement of PMA-induced mucin secretion (∼40%) compared with control cells transfected with no DNA or empty vector (pEGFP-N1). In additional control studies, the effects of Bryostatin 1 on both mucin secretion and phosphorylation of MARCKS were markedly attenuated in cells transfected with the dnPKCδ construct (data not shown). PKCε constructs, either wild type or mutated, did not affect the secretory response to PMA. As shown in Figure 4B, phosphorylation of MARCKS in response to PMA was decreased in HBE1 cells transfected with the dominant-negative PKCδ construct (pEGFP-N1/PKCδK376R) but increased in cells transfected with the wild-type PKCδ construct (pEGFP-N1/PKCδ). PKCε constructs, either wild type or mutated, had no effect on PMA-induced MARCKS phosphorylation when transfected into HBE1 cells (Figure 4C).Figure 4Transient transfection of HBE1 cells with a dominant-negative PKCδ construct results in reduction of mucin hypersecretion. HBE1 cells were transiently transfected with empty vector (pEGFP-N1), a wild-type PKCδ construct (pEGFP-N1/PKCδ), or a dominant-negative construct (pEGFP-N1/PKCδK376R) using the FuGene 6 transfection reagent as described in Materials and Methods. As an additional control, a wild-type (pHACE/PKCε) and a dominant-negative PKCε construct (pHACE/PKCεK437R) were also transfected. A: After 48 hours transfection, cells were exposed to 500 nmol/L PMA (lanes 2 to 8) or vehicle control (lane 1) for 15 minutes, at which time media were collected and mucin secretion assessed by enzyme-linked immunosorbent assay. Data are significantly different from media control (*P < 0.05; **P < 0.001); significantly different from cells transfected with n}, number={6}, journal={AMERICAN JOURNAL OF PATHOLOGY}, author={Park, Jin-Ah and Crews, Anne L. and Lampe, William R. and Fang, Shijing and Park, Joungjoa and Adler, Kenneth B.}, year={2007}, month={Dec}, pages={1822–1830} }
 @article{park_crews_adler_2007, title={Protein kinase C delta regulates airway mucin secretion via phosphorylation of MARCKS protein.}, volume={175}, journal={404nOtfound}, author={Park, J. A. and Crews, A. L. and Adler, K. B.}, year={2007}, pages={A752} }
 @article{raiford_lin_park_fang_crews_adler_2007, title={Proteomic analysis of mucin granule membrane-associated proteins in human airway epithelial cells: a mechanistic link between MARCKS and hClCA1?}, volume={175}, journal={404nOtfound}, author={Raiford, K. L. and Lin, K. W. and Park, J. and Fang, S. and Crews, A. L. and Adler, K. B.}, year={2007}, pages={A511} }
 @article{hawkins_birkenheuer_marr_rogala_large_adler_2007, title={Quantification of mucin gene expression in tracheobronchial epithelium of healthy dogs and dogs with chronic bronchitis}, volume={68}, ISSN={["1943-5681"]}, DOI={10.2460/ajvr.68.4.435}, abstractNote={Abstract}, number={4}, journal={AMERICAN JOURNAL OF VETERINARY RESEARCH}, author={Hawkins, Eleanor C. and Birkenheuer, Adam J. and Marr, Henry S. and Rogala, Allison R. and Large, Edward E. and Adler, Kenneth B.}, year={2007}, month={Apr}, pages={435–440} }
 @article{wang_adler_erjefalt_bai_2007, title={Role of airway epithelial dysfunction in development of acute lung injury and acute respiratory distress syndrome.}, volume={1}, DOI={10.1586/17476348.1.1.149}, abstractNote={Acute lung injury (ALI) and/or acute respiratory distress syndrome (ARDS) are common and important stages of both pulmonary and systemic critical illnesses. ALI/ARDS is categorized as primary or secondary based on the etiology of the disease. There is increasing evidence to suggest the involvement of airway epithelial cells in the pathogenesis of ALI/ARDS. The airway epithelial cell is a new candidate as a biological target responsible for development of the disease and the role of these cells in the pathogenesis of ALI/ARDS is under investigation. This review describes the importance of the airway epithelial cell in the development of ALI/ARDS, its role as the first line of lung defense facing local and primary challenges, its role as an important player in the development of airway inflammation and remodeling, as an inflammatory promoter for initiating both local and systemic inflammation and as an active producer of several inflammatory and anti-inflammatory mediators. It is hypothesized that airway epithelial cells may contribute to ALI/ARDS via Toll-like receptor-involved mediators, reactive oxygen species-involved reactions and an imbalance between protease and antiprotease activation. The airway epithelial cell may be a valuable therapeutic target for discovering and developing new drugs and/or new therapeutic strategies for ALI/ARDS.}, journal={404nOtfound}, author={Wang, X. and Adler, K. B. and Erjefalt, J. and Bai, C.}, year={2007}, pages={149–155} }
 @article{waterman_park_crews_adler_2007, title={The (S)?enantiomer of albuterol activates expression of iNOS via activation of NF-kB in primary cultures of normal human bronchial epithelial cells.}, volume={175}, journal={404nOtfound}, author={Waterman, J. T. and Park, J. A. and Crews, A. L. and Adler, K. B.}, year={2007}, pages={A178} }
 @article{chorley_li_fang_park_adler_2006, title={(R)-Albuterol elicits antiinflammatory effects in human airway epithelial cells via iNOS}, volume={34}, DOI={10.1165/rcmb.2005-03380C}, number={1}, journal={American Journal of Respiratory Cell and Molecular Biology}, author={Chorley, B. N. and Li, Y. H. and Fang, S. J. and Park, J. A. and Adler, K. B.}, year={2006}, pages={119–127} }
 @article{takashi_park_fang_koyama_parikh_adler_2006, title={A peptide against the N-terminus of myristoylated alanine-rich C kinase substrate inhibits degranulation of human leukocytes in vitro}, volume={34}, ISSN={["1535-4989"]}, DOI={10.1165/rcmb.2006-0030RC}, abstractNote={Leukocytes synthesize a variety of inflammatory mediators that are packaged and stored in the cytoplasm within membrane-bound granules. Upon stimulation, the cells secrete the granule contents via an exocytotic process whereby the granules translocate to the cell periphery, the granule membranes fuse with the plasma membrane, and the granule contents are released extracellularly. We have reported previously that another exocytotic process, release of mucin by secretory cells of the airway epithelium, is regulated by the myristoylated alanine-rich C kinase substrate (MARCKS) (Li Y, Martin LD, Spizz G, Adler KB. MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro. J Biol Chem 2001;276:40982-40990; Singer M, Martin LD, Vargaftig BB, Park J, Gruber AD, Li Y, Adler KB. A MARCKS-related peptide blocks mucus hypersecretion in a mouse model of asthma. Nat Med 2004;10:193-196). In those studies, mucin secretion in vitro and in vivo was attenuated by a synthetic peptide identical to the N-terminus of MARCKS, named the MANS peptide (Li and colleagues, 2001). In this study, we used the MANS peptide to investigate possible involvement of MARCKS in secretion of leukocyte granule proteins. In neutrophils isolated from human blood, phorbol 12-myristate 13-acetate-induced myeloperoxidase release was attenuated in a concentration-dependent manner by MANS but not by equal concentrations of a missense control peptide. In additional studies using human leukocyte cell lines, secretion of eosinophil peroxidase from the eosinophil-like cell line HL-60 clone 15, lysozyme from the monocytic leukemia cell line U937, and granzyme from the lymphocyte natural killer cell line NK-92 were attenuated by preincubation of the cells with MANS but not with the missense control peptide. The results indicate that MARCKS protein may play an important role in the secretion of membrane-bound granules from different leukocytes. MARCKS may be an important component of secretory pathways associated with release of granules by different cell types.}, number={6}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Takashi, Shuji and Park, Joungjoa and Fang, Shijing and Koyama, Sekiya and Parikh, Indu and Adler, Kenneth B.}, year={2006}, month={Jun}, pages={647–652} }
 @article{wang_adler_chaudry_ward_2006, title={Better understanding of organ dysfunction requires proteomic involvement}, volume={5}, ISSN={["1535-3893"]}, DOI={10.1021/pr050441n}, abstractNote={Organ dysfunction is defined as a systemic consequence of acute and chronic diseases, a critical and important phase of disease development. The mortality of patients with severe illness is highly correlated with the number and duration of dysfunctional organs. There is still not an efficient and specific therapy to improve the prognosis of patients with organ dysfunction, due to the complexity and severity of the disease. There is a great need to understand molecular mechanisms of the disease, identify disease-related biomarkers, and validate therapeutic effects. Thus, it is important to have a special attention from proteomic scientists to explore the combination between advanced proteomic biotechnology, clinical proteomics, tissue imaging and profiling, and organ dysfunction score systems, to improve the clinical outcomes of these patients.}, number={5}, journal={JOURNAL OF PROTEOME RESEARCH}, author={Wang, XD and Adler, KB and Chaudry, IH and Ward, PA}, year={2006}, month={May}, pages={1060–1062} }
 @article{chorley_crews_li_adler_minnicozzi_martin_2006, title={Differential Muc2 and Muc5ac secretion by stimulated guinea pig tracheal epithelial cells in vitro}, volume={7}, journal={Respiratory Research}, author={Chorley, B. N. and Crews, A. L. and Li, Y. H. and Adler, K. B. and Minnicozzi, M. and Martin, L. D.}, year={2006}, pages={35} }
 @misc{zhao_adler_bai_tang_wang_2006, title={Epithelial proteomics in multiple organs and tissues: Similarities and variations between cells, organs, and diseases}, volume={5}, ISSN={["1535-3907"]}, DOI={10.1021/pr050389v}, abstractNote={Epithelial cells play an important role in physiological and pathophysiological situations, with organ-, tissue-, type-, and function-specific patterns. Proteome analysis has been used to study epithelial-origin diseases and identify novel prognostic, diagnostic, and therapeutic markers. The present review compares the variation of sample preparation for epithelial proteomic analysis, search similarities, and differences of epithelial proteomics between different cells, locations, and diseases. We focus on specificity of proteomic markers for epithelial-involved diseases. Proteomic alterations in epithelial cell lines were mapped to understand protein patterns, differentiation, oncogenesis, and pathogenesis of epithelial-origin diseases. Changes of proteomic patterns depend on different epithelial cell lines, challenges, and preparation. Epithelial protein profiles associated with intracellular locations and protein function. Epithelial proteomics has been greatly developed to link clinical questions, e.g., disease severity, biomarkers for disease diagnosis, and drug targets. There is an exciting and attractive start to link epithelial proteomics with histology of clinical samples. From the present review, we can find that most of disease-associated investigation of epithelial proteomics has been focused on epithelial-origin cancer. There is a significant gap of epithelial proteomics between acute and chronic organ injury, inflammation, and multiple organ dysfunction. Epithelial proteomics will provide powerful information on the relationships between biological molecules and disease mechanisms. Epithelial proteomics strategies and approaches should become more global, multidimensional, and systemic.}, number={4}, journal={JOURNAL OF PROTEOME RESEARCH}, author={Zhao, H and Adler, KB and Bai, CX and Tang, FD and Wang, XD}, year={2006}, month={Apr}, pages={743–755} }
 @article{agrawal_subramannian_adler_ram_ghosh_farim_dickey_2006, title={MARCKS related peptide improves airway obstruction related to mucus secretion in a mouse model of asthma.}, volume={3}, journal={404nOtfound}, author={Agrawal, A. and Subramannian, R. and Adler, K. B. and Ram, A. and Ghosh, B. and Farim, M. and Dickey, B. F.}, year={2006}, pages={A713} }
 @article{adler_fang_lin_park_2006, title={Mechanisms of mucus secretion in the airways.}, volume={2}, DOI={10.1080/17471060500462450}, journal={404nOtfound}, author={Adler, K. B. and Fang, S. and Lin, K-W. and Park, J.}, year={2006}, pages={24–29} }
 @misc{takashi_parikh_adler_martin_y._2006, title={Methods for regulating inflammatory mediators and peptides useful therein}, volume={7,544,772}, number={2006 Sep. 28}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Takashi, S. and Parikh, I. and Adler, K. B. and Martin, L. D. and Y., Li}, year={2006} }
 @article{park_adler_2006, title={Potential therapy for mucus hypersecretion in chronic obstructive pulmonary disease [translated into Chinese].}, volume={86}, journal={404nOtfound}, author={Park, J. A. and Adler, K. B.}, year={2006}, pages={2890–2892} }
 @article{park_fang_adler_2006, title={Regulation of airway mucin secretion by MARCKS protein involves the chaperones heat shock protein 70 and Cysteine string protein.}, volume={3}, DOI={10.1513/pats.200603-067ms}, journal={404nOtfound}, author={Park, J. and Fang, S. and Adler, K. B.}, year={2006}, pages={493a} }
 @article{chorley_adler_2005, title={?2-adrenergic receptor activation with (R)-albuterol attenuates GM-CSF expression via an iNOS-mediated pathway in human bronchial epithelial cells in vitro.}, volume={2}, journal={404nOtfound}, author={Chorley, B. N. and Adler, K. B.}, year={2005}, pages={A757} }
 @article{takashi_pettersen_park_fang_akley_adler_2005, title={A peptide directed against the N-terminus of MARCKS protein attenuates release of myeloperoxidase (MPO) from human and canine neutrophils in vitro.}, volume={2}, journal={404nOtfound}, author={Takashi, S. and Pettersen, C. A. and Park, J. and Fang, S. and Akley, N. J. and Adler, K. B.}, year={2005}, pages={A633} }
 @misc{martin_adler_macchione_akley_mckane_2005, title={Culture system for mouse tracheal epithelial cells}, volume={6,933,149}, number={2005 Aug. 23}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Martin, L. D. and Adler, K. B. and Macchione, M. and Akley, N. J. and McKane, S. A.}, year={2005} }
 @article{park_fang_adler_2005, title={Cysteine String Protein is involved in mucin secretion from normal human bronchial epithelial (NHBE) cells in vitro.}, volume={2}, journal={404nOtfound}, author={Park, J. and Fang, S. and Adler, K. B.}, year={2005}, pages={A215} }
 @article{adler_2005, title={Expert's opinion on potential role of epithelial cells in pathogenesis of organ dysfunction}, volume={1}, ISSN={1747-1060 1747-1079}, url={http://dx.doi.org/10.1080/17471060500219652}, DOI={10.1080/17471060500219652}, number={1}, journal={Journal of Organ Dysfunction}, publisher={Informa UK Limited}, author={Adler, Kenneth B.}, year={2005}, month={Jan}, pages={24–25} }
 @article{park_chorley_adler_2005, title={Human Neutrophil Elastase provokes mucin secretion by NHBE cells via a Protein Kinase C delta (PKC?)?mediated mechanism.}, volume={2}, journal={404nOtfound}, author={Park, J.-A. and Chorley, B. N. and Adler, K. B.}, year={2005}, pages={A110} }
 @article{park_he_martin_li_chorley_adler_2005, title={Human neutrophil elastase induces hypersecretion of mucin from well-differentiated human bronchial epithelial cells in vitro via a protein kinase C delta-mediated mechanism}, volume={167}, ISSN={["1525-2191"]}, DOI={10.1016/S0002-9440(10)62040-8}, abstractNote={The presence of mucus obstruction and neutrophil-predominant inflammation in several lung disorders, such as cystic fibrosis, suggests a relationship between neutrophils and excess mucus production. Mechanisms of human neutrophil elastase (HNE)-induced mucin secretion by well-differentiated normal human bronchial epithelial (NHBE) cells maintained in air/liquid interface culture were investigated. HNE increased mucin secretion in a concentration-dependent manner, with maximal stimulation (more than twofold) occurring within a short (15 minutes) time period. Mucins MUC5AC and MUC5B, but not MUC2, were released in response to HNE. Stimulation of mucin secretion required partial elastase enzymatic activity and did not appear to involve a soluble product released by the cells. HNE-stimulated secretion involved activation of protein kinase C (PKC), as HNE exposure rapidly provoked PKC enzymatic activity that was attenuated by the general PKC inhibitors calphostin C and bisindoylmaleimide I. Of the different isoforms, PKCα, δ, ζ, λ, ι, and ε were constitutively expressed in NHBE cells while PKCβ, η, and μ were PMA-inducible. PKCδ was the only isoform to translocate from cytoplasm to membrane in response to HNE. Inhibition of PKCδ attenuated HNE-mediated mucin secretion. The results suggest HNE stimulation of mucin release by human airway epithelial cells involves intracellular activation of PKC, specifically the δ isoform. The presence of mucus obstruction and neutrophil-predominant inflammation in several lung disorders, such as cystic fibrosis, suggests a relationship between neutrophils and excess mucus production. Mechanisms of human neutrophil elastase (HNE)-induced mucin secretion by well-differentiated normal human bronchial epithelial (NHBE) cells maintained in air/liquid interface culture were investigated. HNE increased mucin secretion in a concentration-dependent manner, with maximal stimulation (more than twofold) occurring within a short (15 minutes) time period. Mucins MUC5AC and MUC5B, but not MUC2, were released in response to HNE. Stimulation of mucin secretion required partial elastase enzymatic activity and did not appear to involve a soluble product released by the cells. HNE-stimulated secretion involved activation of protein kinase C (PKC), as HNE exposure rapidly provoked PKC enzymatic activity that was attenuated by the general PKC inhibitors calphostin C and bisindoylmaleimide I. Of the different isoforms, PKCα, δ, ζ, λ, ι, and ε were constitutively expressed in NHBE cells while PKCβ, η, and μ were PMA-inducible. PKCδ was the only isoform to translocate from cytoplasm to membrane in response to HNE. Inhibition of PKCδ attenuated HNE-mediated mucin secretion. The results suggest HNE stimulation of mucin release by human airway epithelial cells involves intracellular activation of PKC, specifically the δ isoform. Neutrophils are involved in a variety of inflammatory lung disorders including chronic bronchitis, bronchiectasis, cystic fibrosis, and probably asthma. In these diseases, the pathological findings of mucus obstruction and neutrophil-predominant inflammation in airways1Fahy JV Kim KW Liu J Boushey HA Prominent neutrophilic inflammation in sputum from subjects with asthma exacerbation.J Allergy Clin Immunol. 1995; 95: 843-852Abstract Full Text Full Text PDF PubMed Scopus (568) Google Scholar, 2Stockley RA Role of inflammation in respiratory tract infections.Am J Med. 1995; 99: 8S-13SAbstract Full Text PDF PubMed Scopus (47) Google Scholar, 3Welsh MD Adair BM Foster JC Effect of BVD virus infection on alveolar macrophage functions.Vet Immunol Immunopathol. 1995; 46: 195-210Crossref PubMed Scopus (54) Google Scholar, 4Mohapatra NK Cheng PW Parker JC Paradiso AM Yankaskas JR Boucher RC Boat TF Alteration of sulfation of glycoconjugates, but not sulfate transport and intracellular inorganic sulfate content in cystic fibrosis airway epithelial cells.Pediatr Res. 1995; 38: 42-48Crossref PubMed Scopus (28) Google Scholar, 5Fahy JV Schuster A Ueki I Boushey HA Nadel JA Mucus hypersecretion in bronchiectasis. The role of neutrophil proteases.Am Rev Respir Dis. 1992; 146: 1430-1433Crossref PubMed Scopus (109) Google Scholar, 6Stockley RA Hill SL Morrison HM Starkie CM Elastolytic activity of sputum and its relation to purulence and to lung function in patients with bronchiectasis.Thorax. 1984; 39: 408-413Crossref PubMed Scopus (93) Google Scholar suggest a relationship between neutrophil recruitment/infiltration and excess mucus production and secretion. Neutrophils store three proteases that have been implicated in airway mucin secretion: elastase,7Breuer R Christensen TG Lucey EC Stone PJ Snider GL An ultrastructural morphometric analysis of elastase-treated hamster bronchi shows discharge followed by progressive accumulation of secretory granules.Am Rev Respir Dis. 1987; 136: 698-703Crossref PubMed Scopus (46) Google Scholar, 8Nadel JA Protease actions on airway secretions. Relevance to cystic fibrosis.Ann NY Acad Sci. 1991; 624: 286-296Crossref PubMed Scopus (25) Google Scholar, 9Kim KC Wasano K Niles RM Schuster JE Stone PJ Brody JS Human neutrophil elastase releases cell surface mucins from primary cultures of hamster tracheal epithelial cells.Proc Natl Acad Sci USA. 1987; 84: 9304-9308Crossref PubMed Scopus (124) Google Scholar cathepsin G,10Sommerhoff CP Nadel JA Basbaum CB Caughey GH Neutrophil elastase and cathepsin G stimulate secretion from cultured bovine airway gland serous cells.J Clin Invest. 1990; 85: 682-689Crossref PubMed Scopus (285) Google Scholar and proteinase-3.11Rao NV Marshall BC Gray BH Hoidal JR Interaction of secretory leukocyte protease inhibitor with proteinase-3.Am J Respir Cell Mol Biol. 1993; 8: 612-616Crossref PubMed Scopus (60) Google Scholar, 12Renesto P Halbwachs-Mecarelli L Nusbaum P Lesavre P Chignard M Proteinase 3. A neutrophil proteinase with activity on platelets.J Immunol. 1994; 152: 4612-4617PubMed Google Scholar Of these, human neutrophil elastase (HNE), a major component of primary or azurophilic granules,13Bainton DF Ullyot JL Farquhar MG The development of neutrophilic polymorphonuclear leukocytes in human bone marrow.J Exp Med. 1971; 134: 907-934Crossref PubMed Scopus (565) Google Scholar is the most widely studied with regard to enhanced mucus secretion. Levels of HNE are elevated in airways of patients with chronic bronchitis and cystic fibrosis,14Fick Jr, RB Naegel GP Squier SU Wood RE Gee JB Reynolds HY Proteins of the cystic fibrosis respiratory tract. Fragmented immunoglobulin G opsonic antibody causing defective opsonophagocytosis.J Clin Invest. 1984; 74: 236-248Crossref PubMed Scopus (153) Google Scholar and levels in patients' sputum may exceed 100 μg/ml (3.3 × 10−6 mol/L).15Doring G Goldstein W Botzenhart K Kharazmi A Schiotz PO Hoiby N Dasgupta M Elastase from polymorphonuclear leucocytes: a regulatory enzyme in immune complex disease.Clin Exp Immunol. 1986; 64: 597-605PubMed Google Scholar, 16Goldstein W Doring G Lysosomal enzymes from polymorphonuclear leukocytes and proteinase inhibitors in patients with cystic fibrosis.Am Rev Respir Dis. 1986; 134: 49-56PubMed Google Scholar, 17Suter S Schaad UB Tegner H Ohlsson K Desgrandchamps D Waldvogel FA Levels of free granulocyte elastase in bronchial secretions from patients with cystic fibrosis: effect of antimicrobial treatment against Pseudomonas aeruginosa.J Infect Dis. 1986; 153: 902-909Crossref PubMed Scopus (104) Google Scholar Purified HNE has been shown to provoke secretion of mucin by isolated airway epithelial cells and glands from several species.7Breuer R Christensen TG Lucey EC Stone PJ Snider GL An ultrastructural morphometric analysis of elastase-treated hamster bronchi shows discharge followed by progressive accumulation of secretory granules.Am Rev Respir Dis. 1987; 136: 698-703Crossref PubMed Scopus (46) Google Scholar, 8Nadel JA Protease actions on airway secretions. Relevance to cystic fibrosis.Ann NY Acad Sci. 1991; 624: 286-296Crossref PubMed Scopus (25) Google Scholar, 10Sommerhoff CP Nadel JA Basbaum CB Caughey GH Neutrophil elastase and cathepsin G stimulate secretion from cultured bovine airway gland serous cells.J Clin Invest. 1990; 85: 682-689Crossref PubMed Scopus (285) Google Scholar, 18Kim KC Nassiri J Brody JS Mechanisms of airway goblet cell mucin release: studies with cultured tracheal surface epithelial cells.Am J Respir Cell Mol Biol. 1989; 1: 137-143Crossref PubMed Scopus (46) Google Scholar Although there have been suggestions that interactions between HNE and epithelial cell surfaces may be involved in the response,9Kim KC Wasano K Niles RM Schuster JE Stone PJ Brody JS Human neutrophil elastase releases cell surface mucins from primary cultures of hamster tracheal epithelial cells.Proc Natl Acad Sci USA. 1987; 84: 9304-9308Crossref PubMed Scopus (124) Google Scholar, 19Takeyama K Agusti C Ueki I Lausier J Cardell LO Nadel JA Neutrophil-dependent goblet cell degranulation: role of membrane-bound elastase and adhesion molecules.Am J Physiol. 1998; 275: L294-L302PubMed Google Scholar intracellular mechanisms and signaling pathways associated with HNE-induced mucin hypersecretion have not been elucidated. In this study, well-differentiated primary normal human tracheobronchial epithelial (NHBE) cells maintained in vitro in air/liquid interface were exposed to HNE, and the secretory response assessed. Elastase proved to be a potent mucin secretagogue for NHBE cells, eliciting a robust (greater than twofold) increase in mucin secretion within 15 minutes. The mucin gene products released included those of MUC5AC and MUC5B, but not of MUC2. The mechanism appeared to involve activation of protein kinase C (PKC), as HNE exposure rapidly provoked phosphorylation of MARCKS (myristoylated alanine-rich C kinase substrate) protein, a cellular substrate of PKC, and the mucin secretory response to HNE was attenuated by two different PKC inhibitors. Additional studies provided compelling evidence that PKCδ is the specific PKC isoform involved in the secretory pathway. All chemicals were of analytical grade or higher. NHBE cells, bronchial epithelial basal medium, and supplements for air/liquid interface cell cultures were purchased from Cambrex (San Diego, CA). Endotoxin-free HNE purified from human sputum was purchased from Elastin Products Company (EPC, Owensville, MO). Cytotoxicity was evaluated with CytoTox 96 nonradioactive cytotoxicity assay kits obtained from Promega Corp. (Madison, WI). A specific HNE substrate, MeO-SUC-AL-AL-PRO-VAL-PNA, and an HNE inhibitor, chloromethyl ketone-modified tetrapeptide (CMK), also were purchased from EPC and the HNE inhibitor elastatinal was obtained from Calbiochem (La Jolla, CA). 17Q2 pan mucin antibody was purchased from Babco (Richmond, CA) and anti-MUC5AC (45M1) was purchased from Neomarkers (Fremont, CA). A monoclonal antibody (11C1) against human MUC5B was generously provided by Dr. Reen Wu, University of California at Davis, Davis, CA. The epitope for this antibody, which was generated from the secreted mucin of well-differentiated airway epithelial cells, is not known, but by immunohistochemical staining and Western blot analysis, it appears to recognize the MUC5B peptide. A monoclonal antibody that cross reacts with human MUC2, raised against the guinea pig 522-bp gene sequence analogous to the human D4 domain located in the carboxy-terminal region of the Muc2 gene sequence established previously in our laboratory, was used to detect MUC2 mucins.20Li Y Martin LD Minnicozzi M Greenfeder S Fine J Pettersen CA Chorley B Adler KB Enhanced expression of mucin genes in a guinea pig model of allergic asthma.Am J Respir Cell Mol Biol. 2001; 25: 644-651Crossref PubMed Scopus (32) Google Scholar An ImmunoPure (G) IgG purification kit used for purification of antibodies for enzyme-linked immunosorbent assay (ELISA) was from Pierce (Rockford, IL). For Western blot analysis of PKC isoforms expressed in NHBE cells, a PKC sampler kit and E-cadherin antibody were obtained from BD Biosciences (San Jose, CA). Goat anti-PKCζ and mouse anti-α-tubulin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against phosphorylated (ser) PKC substrate and phosphorylated MARCKS were from Cell Signaling Technology (Beverly, MA). Horseradish peroxidase-conjugated goat anti-mouse IgG and donkey anti-goat IgG also were purchased from Santa Cruz Biotechnology. Horseradish peroxidase-conjugated goat anti-rabbit IgG was purchased from Upstate Biotechnology (Lake Placid, NY). Enhanced chemiluminescence development kits and Hyperfilm were from Amersham Pharmacia Biotech (Piscataway, NJ). All PKC-related inhibitors (ie, calphostin C, bisindoylmaleimide, PKC epsilon and zeta inhibitor peptides, rottlerin) were purchased from Calbiochem. A PepTag assay for nonradioactive detection of PKC activity was purchased from Promega. Other chemical reagents were purchased from Sigma-Aldrich (St. Louis, MO). Transwell-Clear culture inserts and high-binding 96-well assay plates were purchased from Corning Inc. (Corning, NY). Primary cultures of NHBE cells were established using an air/liquid interface cell culture system described previously.21Li Y Martin LD Spizz G Adler KB MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro.J Biol Chem. 2001; 276: 40982-40990Crossref PubMed Scopus (155) Google Scholar Briefly, NHBE cells were expanded once and cells collected and frozen in liquid nitrogen (referred to as passage-2 cells). Air/liquid interface cultures of NHBE cells were established on Transwell-Clear culture inserts thin-coated with rat-tail type I collagen. The basic medium used for NHBE cells was a 1:1 mixture of bronchial epithelial basal medium and high glucose (4.5 g/L) Dulbecco's modified Eagle's medium. The complete medium was composed of basic medium containing a final concentration of 0.5 ng/ml human recombinant epidermal growth factor, 0.5 μg/ml hydrocortisone, 5 μg/ml insulin, 10 μg/ml transferrin, 0.5 μg/ml epinephrine, 6.5 ng/ml triiodothyronine, 50 μg/ml gentamicin, and 50 ng/ml amphotericin-B. In addition, the media contained 0.13 mg/ml bovine pituitary extract made according to the protocol of Bertolero and colleagues,22Bertolero F Kaighn ME Gonda MA Saffiotti U Mouse epidermal keratinocytes. Clonal proliferation and response to hormones and growth factors in serum-free medium.Exp Cell Res. 1984; 155: 64-80Crossref PubMed Scopus (61) Google Scholar 5 × 10−8 mol/L all-trans retinoic acid, 1.5 μg/ml bovine serum albumin, and 20 U/ml nystatin. Frozen NHBE cells were recovered and seeded at a density of ∼2 × 104 cells/cm2 onto the apical surface of the inserts. Media were changed the next day, then every other day until the cells reached ∼90% confluence. At this point, the air/liquid interface was established by removing the apical media, whereas basolateral media were changed daily for up to 21 days. A mucin phenotype was observed at ∼14 days in culture (∼7 days in air-liquid interface culture) and cilia were apparent by 18 days in culture. Mucin secretion reached maximal levels at ∼18 days in culture, so cells cultured for ∼18 to 21 days were used for the experiments described below. HNE stock was made as 10 mg/ml (339 μmol/L) in a 1:1 mixture of glycerol and 0.02 mol/L NaOAc, pH 5.0. The stock was diluted into the culture medium to the final concentration indicated. In all studies, the above solvent appropriately diluted was used as a negative control. NHBE cells were exposed to HNE from both apical and basolateral sides for 15 minutes (unless otherwise indicated). At the end of each treatment, apical medium containing the secreted mucin was collected and quantified. Briefly, 0.25 ml of media containing secreted mucin was collected, 0.5 ml of 1 mmol/L dithiothreitol in phosphate-buffered saline (PBS) was added into each well, and the plates were gently agitated and allowed to stand for 3 minutes before the dithiothreitol/PBS plus mucin was collected in the same tube. Finally, 0.5 ml of 10 μmol/L CMK in PBS was added and collected the same way. Approximately 1.25 ml of the collected mucin mixture with dithiothreitol and CMK was centrifuged at 8000 rpm for 5 minutes to remove cell debris, and then collected in a fresh tube. Phenylmethyl sulfonyl fluoride was added to a final concentration of 1 mmol/L. Baseline and treatment mucin secretions were collected from each culture plate. Baseline mucin secretion was collected to normalize variations from well to well, and to control for possible release of mucin in response to the stress of media change or washing. After the baseline mucin secretion sample was collected, the cells were rested overnight and exposed to test agents the next day for indicated periods of time. Mucin samples were quantified using specific ELISA methods. Firstly, total mucin was quantified by a double-sandwich ELISA using a pan-mucin antibody, 17Q2, that cross reacts with a carbohydrate epitope on human mucins, as described previously.21Li Y Martin LD Spizz G Adler KB MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro.J Biol Chem. 2001; 276: 40982-40990Crossref PubMed Scopus (155) Google Scholar Additional studies were performed using ELISAs for secreted protein products of the mucin genes MUC5AC, MUC5B, and MUC2 to determine which mucin gene products were being released on exposure to HNE. MUC5AC was measured via ELISA as described by Takeyama and colleagues23Takeyama K Dabbagh K Lee HM Agusti C Lausier JA Ueki IF Grattan KM Nadel JA Epidermal growth factor system regulates mucin production in airways.Proc Natl Acad Sci USA. 1999; 96: 3081-3086Crossref PubMed Scopus (522) Google Scholar using the 45M1 antibody. MUC5B protein was assayed via a standard double-sandwich ELISA method using the 11C1 monoclonal antibody against MUC5B provided by Dr. Reen Wu, University of California, Davis, Davis, CA, as described previously.24Groneberg DA Eynott PR Oates T Lim S Wu R Carlstedt L Nicholson AG Chung KF Expression of MUC5AC and MUC5B mucins in normal and cystic fibrosis lung.Respir Med. 2002; 96: 81-86Abstract Full Text PDF PubMed Scopus (154) Google Scholar, 25Crowther JR ELISA. Theory and practice.Methods Mol Biol. 1995; 42: 1-218PubMed Google Scholar The MUC2 gene product was quantified by modification of an ELISA as described previously.20Li Y Martin LD Minnicozzi M Greenfeder S Fine J Pettersen CA Chorley B Adler KB Enhanced expression of mucin genes in a guinea pig model of allergic asthma.Am J Respir Cell Mol Biol. 2001; 25: 644-651Crossref PubMed Scopus (32) Google Scholar HNE activity assays were performed following the manufacturer's protocol (EPC). HNE substrate was prepared in substrate buffer (Tris-NaCl buffer: 0.1 mol/L Tris, pH 7.5, containing 0.5 mol/L NaCl and 0.01% Na3N). Briefly, 3 ml of substrate solution at 25°C was added to test tubes, 1.0 μg of HNE then was added, and the developed color was read immediately and continuously thereafter at 1 minute intervals. Elastase activity was reflected by the rate increase in absorbance in time units (minutes). Color development was read at 410 nm on a spectrophotometer UV160U (Shimadzu, Kyoto, Japan). The specific activity of HNE was expressed as U/mg, and results expressed as percentage of activity of native HNE for each treatment. Effects of enzymatic inhibition of HNE were investigated using three different elastase inhibitors: 1) elastatinal, a natural HNE inhibitor produced by Actinomycetes;26Umezawa H Structures and activities of protease inhibitors of microbial origin.Methods Enzymol. 1976; 45: 678-695Crossref PubMed Scopus (303) Google Scholar 2) CMK, a synthetic tetrapeptide;27Rees DD Brain JD Wohl ME Humes JL Mumford RA Inhibition of neutrophil elastase in CF sputum by L-658,758.J Pharmacol Exp Ther. 1997; 283: 1201-1206PubMed Google Scholar and 3) α1-antitrypsin (α1-AT), a physiological HNE inhibitor.28Gadek JE Fells GA Zimmerman RL Rennard SI Crystal RG Antielastases of the human alveolar structures. Implications for the protease-antiprotease theory of emphysema.J Clin Invest. 1981; 68: 889-898Crossref PubMed Scopus (311) Google Scholar The inhibitors were added directly to HNE, incubated for 15 minutes at 37°C, and then added directly to the cells for another 15 minutes. At the end of this exposure, secreted mucin was collected and quantified as described above. To determine whether HNE enzymatic activity was directly required for stimulated mucin secretion, or if a secondary product(s) released by NHBE cells after exposure to HNE could be involved in the secretory response, NHBE cells were exposed to HNE (or vehicle) for 5 minutes. After exposure, the conditioned medium was collected and treated with 5 μmol/L of the HNE enzymatic inhibitor, α1-AT, for 15 minutes, at which time this α1-AT-treated medium was added to a new set of NHBE cells and effects on mucin secretion quantified as described above. The PKC inhibitors, bisindolylmaleimide I (10, 100, 1000 nmol/L)29Martiny-Baron G Kazanietz MG Mischak H Blumberg PM Kochs G Hug H Marme D Schachtele C Selective inhibition of protein kinase C isozymes by the indolocarbazole Go 6976.J Biol Chem. 1993; 268: 9194-9197Abstract Full Text PDF PubMed Google Scholar or calphostin C (5, 50, 500 nmol/L)30Takahashi I Saitoh Y Yoshida M Sano H Nakano H Morimoto M Tamaoki T UCN-01 and UCN-02, new selective inhibitors of protein kinase C. II. Purification, physico-chemical properties, structural determination and biological activities.J Antibiot (Tokyo). 1989; 42: 571-576Crossref PubMed Scopus (168) Google Scholar were used to determine PKC involvement in HNE-induced mucin secretion. NHBE cells were preincubated with these agents (or vehicle control) for 15 minutes, then HNE was added for another 15 minutes before mucin secretion was quantified as described above. PKC activity in NHBE cells after exposure to HNE was assessed using a PepTag assay for nonradioactive detection of PKC (following the manufacturer's protocol). Briefly, 10 μg of protein extracted from each treatment of NHBE cells was added into the PKC reaction buffer (20 mmol/L HEPES, pH 7.4, 1.3 mmol/L CaCl2, 1 mmol/L dithiothreitol, 10 mmol/L MgCl2, 1 mmol/L ATP) containing 1 mg/ml phosphatidylserine and PepTag C1 PKC substrate peptide (P-L-S-R-T-L-S-V-A-A-K) conjugated with fluorescent dye, and incubated for 30 minutes at 30°C. The reaction was stopped by boiling at 100°C for 10 minutes. Reaction mixtures were separated on 0.8% agarose gels and proteins quantified by Labworks image acquisition and analysis software (UVP Bioimaging System, Upland, CA). Phosphorylation of MARCKS was detected by Western blot using an antibody against phophospecific-MARCKS. After treatments, NHBE cells were washed with ice-cold PBS twice and then scraped into lysis buffer (50 mmol/L Tris, pH 7.5, 1 mmol/L ethylenediamine tetraacetic acid, 100 mmol/L NaCl, 1 mmol/L phenylmethyl sulfonyl fluoride) using a rubber policemen. The collected cells were lysed by sonication. For separation of cytosolic and membrane fractions, the lysates were spun at 400,000 × g in a Sorvall Discovery 100S ultracentrifuge (Sorvall, Inc. Newtown, CT) for 1 hour. The supernatant was reserved as the cytosolic sample. The pellet was resuspended in the same lysis buffer containing 0.05% Triton-100, dissolved by sonication, and incubated on ice for 30 minutes. After incubation, the same ultracentrifugation as described above was performed on the pellet mixture, and the supernatant separated from the pellet mixture was reserved as the membrane fraction. For preparation of whole cell crude lysates, the disrupted cellular mixture was centrifuged at 15,000 rpm in an Eppendorf 5417R centrifuge (Eppendorf Corp., Hamburg, Germany) for 1 hour at 4°C. The supernatant was collected as the whole crude NHBE cell lysate. The protein concentration of cell lysate samples was quantified by a Bradford assay (Bio-Rad Laboratories, Hercules, CA). Each sample was boiled in 2× sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer for 10 minutes, loaded on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, and transferred to a polyvinylidene difluoride membrane (Micron Separation Inc., Westborough, MA). After blocking with 5% skim milk, the antigen was captured by the specific PKC antibody and further amplified by binding to horseradish peroxidase-conjugated anti-mouse or anti-rabbit antibodies. Anti-α-tubulin and E-cadherin antibodies were used for cytosolic and membrane controls, respectively, for each sample. Final development was accomplished by the enhanced chemiluminescence method. The amount of each PKC isoform was analyzed by Labworks image acquisition and analysis software. Because the studies above indicated that PKCδ was the only isoform to translocate to membranes in response to HNE, additional studies were performed with rottlerin, an inhibitor of PKCδ and θ.31Gschwendt M Muller HJ Kielbassa K Zang R Kittstein W Rincke G Marks F Rottlerin, a novel protein kinase inhibitor.Biochem Biophys Res Commun. 1994; 199: 93-98Crossref PubMed Scopus (767) Google Scholar (Because PKCθ was not expressed in NHBE cells under basal or stimulated conditions, rottlerin is referred to below as a specific inhibitor of PKCδ). Rottlerin has the following potency against PKC isoforms: PKC δ (IC50 = 3 to 6 μmol/L); PKCθ (IC50 = 50 μmol/L); PKCα, PKCβ, and PKC γ (IC50 = 30 to 42 μmol/L); PKCε, PKCη, and PKCζ (IC50 = 80 to 100 μmol/L). It also can inhibit CaM kinase III (IC50 = 5.3 μmol/L).31Gschwendt M Muller HJ Kielbassa K Zang R Kittstein W Rincke G Marks F Rottlerin, a novel protein kinase inhibitor.Biochem Biophys Res Commun. 1994; 199: 93-98Crossref PubMed Scopus (767) Google Scholar, 32Villalba M Kasibhatla S Genestier L Mahboubi A Green DR Altman A Protein kinase C cooperates with calcineurin to induce fas ligand expression during activation-induced T cell death.J Immunol. 1999; 163: 5813-5819PubMed Google Scholar Cells were preincubated with rottlerin (1.5 μmol/L; IC50 = 3 to 6 μmol/L) for 20 minutes before exposure to HNE, and effects on PKC activity [using detection of phosphorylated (ser) PKC substrate] and on HNE-induced mucin secretion were assessed. As additional controls, the potential role of other PKC isoforms present in these cells was assessed: cells were exposed to the following specific inhibitors for 15 minutes before exposure to HNE and assay for mucin secretion: The PKCα/β inhibitor, Gö 6976 (10 nmol/L; IC50 = 2 ∼ 6 nmol/L);29Martiny-Baron G Kazanietz MG Mischak H Blumberg PM Kochs G Hug H Marme D Schachtele C Selective inhibition of protein kinase C isozymes by the indolocarbazole Go 6976.J Biol Chem. 1993; 268: 9194-9197Abstract Full Text PDF PubMed Google Scholar a PKCζ peptide inhibitor (50 μmol/L; Ser-Ile-Tyr-Arg-Arg-Gly-Ala-Arg-Arg-Trp-Arg-Lys-Leu; IC50 = 10 μmol/L);33Bandyopadhyay G Standaert ML Galloway L Moscat J Farese RV Evidence for involvement of protein kinase C (PKC)-zeta and noninvolvement of diacylglycerol-sensitive PKCs in insulin-stimulated glucose transport in L6 myotubes.Endocrinology. 1997; 138: 4721-4731Crossref PubMed Scopus (210) Google Scholar or a PKCε peptide inhibitor (3 ∼ 300 μmol/L; Glu-Ala-Val-Ser-Leu-Lys-Pro-Thr; IC50 = 80.3 μmol/L).34Johnson JA Gray MO Chen CH Mochly-Rosen D A protein kinase C translocation inhibitor as an isozyme-selective antagonist of cardiac function.J Biol Chem. 1996; 271: 24962-24966Crossref PubMed Scopus (343) Google Scholar, 35Mendez CF Leibiger IB Leibiger B Hoy M Gromada J Berggren PO Bertorello AM Rapid association of protein kinase C-epsilon with insulin granules is essential for insulin exocytosis.J Biol Chem. 2003; 278: 44753-44757Crossref PubMed Scopus (59) Google Scholar Data were expressed as the ratio of treatment to the corresponding vehicle control. Results were evaluated using one-way analysis of variance with Bonferroni posttest correction for multiple comparisons.36Kleinbaum DG Kupper LL Muller KE Applied Regression Analysis and Other Multivariable Methods. PWS-Kent Pub. Co., Boston1988: 341-386Google Scholar A P value of <0.05 was considered significant. All reagents used were tested for cytotoxicity using a Promega Cytotox 96 nonradioactive cytotoxicity assay kit according to the manufacturer's instructions. The data were expressed as the ratio of released lactate dehydrogenase to total lactate dehydrogenase. Released lactate dehydrogenase never exceeded 10% of total lactate dehydrogenase (data not shown) in any of the experiments below. As illustrated in Figure 1, HNE stimulated mucin secretion by NHBE cells. Maximal mucin secretion was elicited after 15 minutes exposure to HNE (Figure 1A) so this time point was chosen for additional experiments. HNE increased mucin secretion in a concentration-dependent manner, with 0.01 to 1.0 μmol/L HNE increasing secretion significantly over vehicle control (Figure 1B). Secretion of major gel-forming mucins, including MUC2, MUC5AC, and MUC5B, was investigated after exposure to HNE. As illustrated in Figure 2, HNE enhanced release of both MUC5AC and MUC5B mucins from NHBE cells in a concentration-dependent manner. Secretion of MUC2 mucin was significantly decreased by HNE. Elastatinal appeared to be the weakest of the three HNE inhibitors used in this study because the highest concentration used, 100 μmol/L, blocked only 50% of HNE enzymatic activity and did not affect HNE-stimulated mucin secretion (Figure 3A). CMK proved to be a more potent HNE enzymatic inhibitor because 50 μmol/L CMK completely blocked the enzymatic activity of 1 μmol/L HNE, whereas lower concentrations partially inhibited HNE activity in a concentration-dependent manner. CMK also showed an inhibitory effect on HNE-stimulated mucin secretion in a concentration-dependent manner w}, number={3}, journal={AMERICAN JOURNAL OF PATHOLOGY}, author={Park, JA and He, F and Martin, LD and Li, YH and Chorley, BN and Adler, KB}, year={2005}, month={Sep}, pages={651–661} }
 @article{singer_martin_vargaftig_park_gruber_li_adler_2004, title={A MARCKS-related peptide blocks mucus hypersecretion in a mouse model of asthma}, volume={10}, ISSN={["1546-170X"]}, DOI={10.1038/nm983}, abstractNote={Mucus hypersecretion is a crucial feature of pulmonary diseases such as asthma, chronic bronchitis and cystic fibrosis. Despite much research, there is still no effective therapy for this condition. Recently, we showed that the myristoylated, alanine-rich C-kinase substrate (MARCKS) protein is required for mucus secretion by human bronchial epithelial cells in culture. Having synthesized a peptide corresponding to the N-terminal domain of MARCKS, we now show that the intratracheal instillation of this peptide blocks mucus hypersecretion in a mouse model of asthma. A missense peptide with the same amino acid composition has no effect. Based on quantitative histochemical analysis of the mouse airways, the peptide seems to act by blocking mucus release from goblet cells, possibly by inhibiting the attachment of MARCKS to membranes of intracellular mucin granules. These results support a pivotal role for MARCKS protein, specifically its N-terminal region, in modulating this secretory process in mammalian airways. Intratracheal administration of this MARCKS-related peptide could therapeutically reduce mucus secretion in the airways of human patients with asthma, chronic bronchitis and cystic fibrosis.}, number={2}, journal={NATURE MEDICINE}, author={Singer, M and Martin, LD and Vargaftig, BB and Park, J and Gruber, AD and Li, YH and Adler, KB}, year={2004}, month={Feb}, pages={193–196} }
 @misc{martin_adler_li_2004, title={Blocking peptide for inflammatory cell secretion}, volume={WO/2003/000027}, number={2004 Sep. 16}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Martin, L. D. and Adler, K. B. and Li, Y}, year={2004} }
 @article{chorley_adler_2004, title={Inducible nitric oxide synthase/protein kinase G signaling pathway suppresses granulocyte macrophage colony stimulating factor transcription in normal human bronchial epithelial cells in vitro}, volume={169}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Chorley, B. N. and Adler, K. B.}, year={2004}, pages={A421} }
 @article{park_fang_gruber_adler_2004, title={MARCKS protein interaction with the ?secretory module? regulates airway mucin secretion.}, volume={169}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Park, J. and Fang, S. and Gruber, A. D. and Adler, K. B.}, year={2004}, pages={A535} }
 @article{hashimoto_graham_ho_adler_collins_olson_zhou_suzutani_jones_goleniewska_et al._2004, title={Respiratory syncytial virus in allergic lung inflammation increases Muc5ac and Gob-5}, volume={170}, ISSN={["1535-4970"]}, DOI={10.1164/rccm.200301-030OC}, abstractNote={Respiratory syncytial virus (RSV) is associated with wheezing and childhood asthma. We previously reported that RSV infection prolongs methacholine-induced airway hyperresponsiveness in ovalbumin (OVA)-sensitized mice. In addition, allergically sensitized RSV-infected (OVA/RSV) mice had more abundant airway epithelial mucus production compared with OVA mice 14 days after infection, whereas there was almost no mucus in mice that were only RSV infected. We hypothesized that this increased mucus was associated with mucosal expression of Muc5ac, a mucus gene expression in airways, and gob-5, a member of the Ca(2)(+)-activated chloride channel family. By histochemical analysis, we found that there was significantly increased staining for gob-5 and Muc5ac in the airways of OVA/RSV mice compared with either OVA mice or allergically sensitized mice that were challenged with inactivated RSV, and virtually no detectable staining in the RSV group. These findings were confirmed by Western blot analysis. The increased mucus expression in the OVA/RSV group was associated with increased lung levels of interleukin-17, a factor known to stimulate airway mucin gene expression. The impact of virus infection combined with allergic inflammation on mucus production may partially explain the more severe disease and airway hyperresponsiveness associated with RSV in the setting of atopy.}, number={3}, journal={AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE}, author={Hashimoto, K and Graham, BS and Ho, SB and Adler, KB and Collins, RD and Olson, SJ and Zhou, WS and Suzutani, T and Jones, PW and Goleniewska, K and et al.}, year={2004}, month={Aug}, pages={306–312} }
 @article{fang_park_adler_2004, title={Small interfering RNA?s directed against MARCKS protein and Heat Shock Protein 70 (Hsp70) attenuate mucin secretion in human airway epithelial cells in vitro.}, volume={169}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Fang, S. and Park, J. and Adler, K. B.}, year={2004}, pages={A712} }
 @article{vargaftig_singer_martin_li_adler_2003, title={A myristoylated peptide directed against the N-terminal region of MARCKS protein inhibits mucin secretion in ovalbumin sensitized/challenged mice in vivo.}, volume={167}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Vargaftig, B. and Singer, M. and Martin, L. D. and Li, Y. and Adler, K. B.}, year={2003}, pages={A17} }
 @article{lin_park_li_adler_2003, title={Activation of protease-activated receptors?2 (PAR-2) is not associated with enhanced mucin secretion by well-differentiated normal human bronchial epithelial cells in vitro.}, volume={167}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Lin, K. W. and Park, J. J. and Li, Y. and Adler, K. B.}, year={2003}, pages={A204} }
 @article{chorley_martin_crews_li_adler_2003, title={Differential effects of albuterol isomers on normal human bronchial epithelial cells in vitro.}, volume={167}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Chorley, B. N. and Martin, L. D. and Crews, A. C. and Li, Y. and Adler, K. B.}, year={2003}, pages={A205} }
 @article{krunkosky_martin_fischer_voynow_adler_2003, title={Effects of TNF alpha on expression of ICAM-1 in human airway epithelial cells in vitro: Oxidant-mediated pathways and transcription factors}, volume={35}, ISSN={["1873-4596"]}, DOI={10.1016/S0891-5849(03)00498-2}, abstractNote={We demonstrate that two different cell-permeable antioxidants, pyrrolidine dithiocarbamate (PDTC) and dimethylthiourea (DMTU), inhibit TNFalpha-induced ICAM-1 surface and gene expression in primary cultures of differentiated normal human bronchial epithelial (NHBE) cells. In addition, TNFalpha stimulates binding of nuclear proteins to the nuclear factor kappa beta (NFkappaB) and the CAAT/enhancer binding protein (C/EBP) consensus sites in the ICAM-1 promoter in these cells. Because these transcription factors have been suggested to be oxidant-sensitive and important in ICAM-1 expression, the potential involvement of reactive oxygen species (ROS) in the response to TNFalpha was investigated. Interestingly, neither PDTC nor DMTU altered binding of NFkappaB complexes. In contrast, either the proteasome inhibitor carbobenzoxy-L-leucy-L-leucy-L-leucinal (MG 132) or the IkappaBalpha inhibitor BAY 11-7082 ablated TNFalpha-induced ICAM-1 gene expression and MG132 inhibited TNFalpha-induced NFkappaB complexes. Surprisingly, either PDTC or DMTU inhibited the binding of TNFalpha-enhanced C/EBP complexes to the consensus site directly adjacent to the NFkappaB site. These results suggest that although TNFalpha enhances binding of C/EBP and NFkappaB complexes in NHBE cells, C/EBP binding seems to involve an oxidant-dependent mechanism, whereas activation of NFkappaB complexes utilizes the ubiquitin-proteasome pathway, a mechanism that seems to be unaltered by the presence of antioxidants. Because interference with either signaling pathway abrogates TNFalpha-induced ICAM-1 expression, activation of both complexes seems to be involved in this response to TNFalpha, but this activation occurs via different intracellular pathways.}, number={9}, journal={FREE RADICAL BIOLOGY AND MEDICINE}, author={Krunkosky, TM and Martin, LD and Fischer, BM and Voynow, JA and Adler, KB}, year={2003}, month={Nov}, pages={1158–1167} }
 @article{park_he_li_martin_adler_2003, title={Human neutrophil elastase provokes release of MUC5B mucin from normal bronchial epithelial cells in vitro via a PKC-dependent mechanism.}, volume={167}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Park, J. A. and He, F. and Li, Y. and Martin, L. D. and Adler, K. B.}, year={2003}, pages={A203} }
 @article{singh_daniels_winsett_richards_doerfler_hatch_adler_gilmour_2003, title={Phenotypic comparison of allergic airway responses to house dust mite in three rat strains}, volume={284}, ISSN={["1522-1504"]}, DOI={10.1152/ajplung.00287.2002}, abstractNote={Brown Norway (BN) rats develop a robust response to antigens in the lung, characterized by a large increase in allergen-specific immune function and pulmonary eosinophilia. The objective of this study was to investigate alternative models by determining whether other rat strains could be sensitized to house dust mite (HDM) antigen and whether the allergic disease process could be worsened with repeated allergen exposure. In general, BN rats sensitized by either subcutaneous or intratracheal routes exhibited increased pulmonary allergy compared with Sprague-Dawley (SD) and Lewis (L) rats. Multiple intratracheal allergen exposures incrementally increased HDM-specific immune function in BN rats but progressively decreased eosinophil recruitment and markers of lung injury. SD rats had more moderate responses, whereas L rats were relatively unresponsive. Because BN rats developed stronger clinical hallmarks of allergic asthma under various immunization regimes compared with SD and L rats, we conclude that the BN is the most appropriate strain for studying allergic asthma-like responses in rats. Phenotypic differences in response to HDM were associated with differences in the Th1/Th2 cytokine balance and antioxidant capacity.}, number={4}, journal={AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY}, author={Singh, P and Daniels, M and Winsett, DW and Richards, J and Doerfler, D and Hatch, G and Adler, KB and Gilmour, MI}, year={2003}, month={Apr}, pages={L588–L598} }
 @article{booth_newcomb_mckane_crews_adler_bonner_martin_2003, title={Proliferation of the airway epithelium in asthma - Are inflammatory cells required?}, volume={123}, ISSN={["0012-3692"]}, DOI={10.1378/chest.123.3_suppl.384S}, abstractNote={Asthma is associated with a T helper type 2 phenotype in which interleukin (IL)-4, IL-5, and IL-13 predominate. In addition, the long-term presence of these inflammatory mediators is thought to lead to airway structural changes that are collectively referred to as airway remodeling. Data from our laboratory, and those of others, have suggested a role for IL-13 in the development of mucous cell hyperplasia that is associated with such remodeling. Others also have suggested a role for inflammatory cells such as neutrophils in mediating this process. Using normal human bronchial epithelial (NHBE) cells differentiated in vitro, we have shown recently that IL-13 (10 ng/mL for 24 h) induces the proliferation of NHBE cells via a mechanism that is dependent on the IL-13-induced release of transforming growth factor (TGF)-α by the epithelial cells. This epithelium-derived TGF-α then acts in an autocrine/paracrine manner to bind the epidermal growth factor receptor (EGFR) on these NHBE cells, enhancing proliferation. Specifically, soluble TGF-α is released by NHBE cells in response to IL-13 exposure (1 h), and the immunohistochemical analysis of cells exposed to IL-13 (after 1 and 4 h) has revealed a lack of membrane-bound TGF-α when compared to control cells. The IL-13-induced proliferative response can be blocked in a concentration-dependent manner by AG1478 (0.1, 1, and 5 μg/mL), which is a specific inhibitor of EGFR tyrosine kinase activity, and is eliminated by neutralizing TGF-α antibodies, while control antibodies (ie, anti-platelet-derived growth factor, epidermal growth factor [EGF], and heparin-binding EGF) have no effect.}, number={3}, journal={CHEST}, author={Booth, BW and Newcomb, DC and McKane, SA and Crews, AL and Adler, KB and Bonner, JC and Martin, LD}, year={2003}, month={Mar}, pages={384S–385S} }
 @article{hashimoto_graham_ho_adler_collins_jones_jarzecka_furlong_peebles_2003, title={Respiratory syncytial virus infection upregulates Muc5ac and gob-5 lung protein expression only in the setting of allergic inflammation.}, volume={167}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Hashimoto, K. and Graham, B. S. and Ho, S. B. and Adler, K. B. and Collins, R. D. and Jones, P. W. and Jarzecka, K. and Furlong, J. and Peebles, R. S., Jr.}, year={2003}, pages={A425} }
 @article{singh_dick_boykin_andrews_adler_gilmour_2003, title={Role of oxidative stress and p38MAP kinase in murine pulmonary responses to LPS and diesel exhaust particles.}, volume={167}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Singh, P. and Dick, C. A. and Boykin, E. and Andrews, D. and Adler, K. B. and Gilmour, I.}, year={2003}, pages={A622} }
 @article{pettersen_adler_2002, title={Airways inflammation and COPD - Epithelial-neutrophil interactions}, volume={121}, ISSN={["1931-3543"]}, DOI={10.1378/chest.121.5_suppl.142S}, abstractNote={Neutrophils are recognized as major cellular mediators of inflammation. They contain specific and highly regulated mechanisms for controlling the expression of adhesion molecules that allow for their tethering and migration into inflammatory sites. These adhesion molecules not only are activated by exogenous pollutants but are regulated by endothelial and epithelial cell signals. Lipid mediators, such as platelet-activating factor, reactive oxygen and nitrogen species, and cytokines from airway epithelial cells, further control neutrophil functions such as infiltration and activation resulting in an increase in respiratory burst activity and release of granule enzymes, such as elastase. Furthermore, virus and bacteria products affect inflammation by increasing secondary epithelial mediators. However, once the endogenous or exogenous agents are expelled, neutrophil populations are programmed to die and are cleared by macrophage phagocytosis.}, number={5}, journal={CHEST}, author={Pettersen, CA and Adler, KB}, year={2002}, month={May}, pages={142S–150S} }
 @article{singh_daniels_winsett_richards_bishop_dick_madden_adler_gilmour_2002, title={Diesel exhaust particles increase lung injury and inflammation during early sensitization in brown Norway rats.}, volume={66}, number={1-S}, journal={Toxicological Sciences}, author={Singh, P. and Daniels, M. J. and Winsett, D. and Richards, J. and Bishop, L. and Dick, C. A. J. and Madden, M. and Adler, K. B. and Gilmour, M. I.}, year={2002}, pages={147} }
 @article{singh_winsett_daniels_dick_adler_gilmour_2002, title={Effects of allergic airways disease on influenza virus infection in brown Norway rats.}, volume={165}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Singh, P. and Winsett, D. W. and Daniels, M. J. and Dick, C. A. J. and Adler, K. B. and Gilmour, M. I.}, year={2002}, pages={A183} }
 @article{booth_adler_martin_2002, title={IL-13 induces intracellular translocation and release of TGF? in human bronchial epithelial cells in vitro.}, volume={165}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Booth, B. W. and Adler, K. B. and Martin, L. D.}, year={2002}, pages={A814} }
 @article{martin_adler_akley_crews_sharova_2002, title={Secretion-competent mouse tracheal epithelial cell culture from the genetically altered mouse - Pathway analysis via gene array}, volume={121}, DOI={10.1016/S0012-3692(15)35478-7}, abstractNote={The ability to create knockout and transgenic mice with phenotypes mimicking a variety of lung diseases has led to a large body of knowledge detailing the role of various gene products in the development of these diseases. Similarly, the use of well-differentiated human airway epithelial cell cultures has led to an understanding of precise signaling pathways regulating cellular functions such as mucus secretion, adhesion molecule and cytokine expression, and epithelial cell proliferation. The ability to combine these two powerful research approaches lies with creating an in vitro mouse tracheal epithelial (MTE) cell culture system. Here, we report the development of such a primary cell system that maintains morphologic and functional characteristics of the in vivo mouse airway epithelium. Specifically, epithelial cells dissociated from intact mouse tracheas are grown in air/liquid interface culture in defined media with or without serum. Under both conditions, Alcian blue/periodic acid-Schiff–positive mucous cells are observed. In contrast, ciliary development appears to require serum, suggesting that it may be possible to further manipulate this cell culture system to allow precise study of either mucous or ciliated cell development. This cell culture system has been examined to ensure its epithelial nature as indicated by Western blot analyses showing the culture findings to be positive for cytokeratin 5 expression. Using a mouse mucin 5ac-specific antibody to detect secreted protein by enzyme-linked immunosorbent assay, the cultures are found to secrete mucin constitutively and in a stimulated manner in response to known secretagogues (phorbol 12-myristate 13-acetate and 8-Br-cyclic guanosine monophosphate). Although a single trachea yields only 1 cm2 of differentiated culture, our preliminary studies indicate sufficient material can be obtained to perform gene array analyses of control and interleukin-13–exposed MTE cell cultures. Thus, we anticipate use of the MTE cell culture system not only to determine specific signaling pathways important to airway epithelial cell changes during lung disease, but by employing cells from knockout and transgenic mice, we expect to obtain an understanding of how expression of genes controlling these pathways is altered by genetic changes. In this manner, it should be possible to directly interface in vitro experimentation to define precise signaling pathways in airway epithelial cells with in vivo whole animal studies.}, number={3}, journal={Chest}, author={Martin, L. D. and Adler, K. B. and Akley, N. J. and Crews, A. and Sharova, L.}, year={2002}, pages={79S} }
 @article{adler_li_2001, title={Airway epithelium and mucus - Intracellular signaling pathways for gene expression and secretion}, volume={25}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.25.4.f214}, abstractNote={It is the rare scientific paper dealing with any aspect of airway mucus that does not open with a statement about the contribution of excess mucus to the pathogenesis of airway obstruction, susceptibility to infection, or compromised defense in a myriad of inflammatory airway diseases, such as chronic bronchitis, asthma, bronchiectasis, or cystic fibrosis. Excess mucus in the airways can result from any of three different lesions, and in most cases various combinations of these: ( 1 ) enhanced production through overexpression of mucin (MUC) genes; ( 2 ) excess production secondary to mucus cell hyperplasia, hypertrophy, or even metaplasia; or ( 3 ) hypersecretion of formed and stored mucin by goblet cells or glands in the airways. In context of a perspective, it may be instructive to trace the historical pathways that have led to our present understanding of the mechanisms associated with mucus-related phenomena. Clearly, the importance of studying production and secretion of mucus (or its glycoprotein component, mucin) was not lost on early researchers. In the 1960s and early 1970s, several groups looked at mucus production and secretion in the airways. However, lack of appropriate in vitro or in vivo model systems made these early studies mostly descriptive and limited, for the most part, to characterization of alcian blue/PAS-stained cells in different regions of the airways in health and disease (1-4). In the mid-1970s, with the introduction of organ culture techniques to study isolated rings or explants of bronchi or trachea from several species, it became possible to investigate mechanisms related to production and secretion of mucin (5, 6). Unfortunately, there were serious problems with explant cultures, not the least of which was quantification of produced or released mucin. The “state-of-the-art” at that time was either to measure carbohydrate components of secreted or retained mucin in the explants (such as sialic acid; fucose, or glucosamine [7]) or to incubate the explants with a radiolabeled sugar (such as tritiated glucosamine) for a time period allowing for incorporation of the label into the mucin glycoproteins, and then measure the released radiolabeled activity as a reflection of secreted mucin, or radioactivity within the tissue as a measure of mucin synthesis (8). For greater specificity, the homogenate or spent medium was either precipitated with trichloroacetic acid, sometimes with the addition of phosphotungstic acid, prior to counting of radioactivity. More accurate quantification was achieved by treatment of the homogenate or spent medium with enzymes to digest other contaminating sugar-containing proteins, such as hyaluronic acid or chondroitin sulfate, with hyaluronidase or chondroitinase ABC, respectively. Separation via column chromatography also improved detection, as the high molecular weight mucins would appear in the void volume (9). A second problem related to organ cultures was the large number of cell types present in the explants, confounding interpretation of effects of added agents on epithelium and making it difficult to attribute responses to any particular cell type. There were some major advancements in the field of mucin research during the 1980s, both in the development of better cell culture techniques and in detection of intraand extracellular (secreted) mucin. The first was a great improvement in our ability to culture cells from airway epithelium from several species. Prior to this time, it was difficult to culture airway epithelial cells so as to maintain differentiated characteristics in vitro , but the development of defined, serum-free medium, as well as improvements in the types of substrata beneath the cultured cells, gave researchers the ability to culture airway epithelial cells that looked and acted somewhat like their in vivo counterparts. Maintaining cells in a defined medium atop a collagen gel provided improved model systems, and in the latter part of the 1980s, the concept of air/liquid interface culture was first introduced. Starting with guinea pig tracheal epithelial cells (10, 11), it was discovered that cells grown on a collagen substrate, atop a permeant filter, with all medium placed beneath the cells and only a humidified air environment above, would result in well-differentiated epithelial cells essentially identical in structure and function to airway epithelium in situ . In quick succession, techniques for air/liquid interface culture of airway epithelium from rat (12), bovine (13), canine (14), primate (15), and eventually human (16-18) cells were developed. At present, culturing human airway epithelial cells in air/liquid interface provides a model system in which the epithelial cells are similar if not identical to human airway epithelium in vivo , and such cells now can be purchased commercially. With regard to detection of intracellular or secreted ( Received in original form September 10, 2001 )}, number={4}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Adler, KB and Li, YH}, year={2001}, month={Oct}, pages={397–400} }
 @article{booth_bonner_adler_martin_2001, title={Autocrine production of TGFa mediates interleukin 13-induced proliferation of human airway epithelial cells during development of a mucous phenotype in vitro.}, volume={163}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Booth, B. and Bonner, J. C. and Adler, K. B. and Martin, L. D.}, year={2001}, pages={A738} }
 @article{macchione_akley_adler_martin_2001, title={Differentiation of murine tracheal epithelial cells in vitro.}, volume={163}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Macchione, M. and Akley, N. J. and Adler, K. B. and Martin, L. D.}, year={2001}, pages={A225} }
 @article{li_martin_minnicozzi_greenfeder_fine_pettersen_chorley_adler_2001, title={Enhanced expression of mucin genes in a guinea pig model of allergic asthma}, volume={25}, ISSN={["1535-4989"]}, DOI={10.1165/ajrcmb.25.5.4485}, abstractNote={The ovalbumin (OVA)-sensitized guinea pig is often used as an animal model of asthma and airway hyperreactivity. A characteristic lesion of asthma is excessive production of mucin in the airways. Mechanistic studies of this lesion in guinea pigs have been limited due to lack of mucin gene probes for this species. The aim of the present study was to clone the cDNAs encoding two major airway mucins (Muc2 and Muc5ac) from the guinea pig, and investigate mucin gene expression in lungs of sensitized animals in response to antigen challenge. We isolated and sequenced two cDNA fragments coding for the sequences located within the carboxyl-terminal cysteine-rich region of guinea pig Muc2 and Muc5ac mucins. Comparison of cloned cDNAs with those from other species revealed high degrees of sequence identity and conservation of all cysteine residues in deduced primary sequences. Based on the resultant sequence information, we also designed oligonucleotide primers for specific detection of guinea-pig Muc2 and Muc5ac steady-state mRNA levels via reverse transcriptase/ polymerase chain reaction (RT-PCR). Levels of both Muc2 and Muc5ac mRNA in lungs of OVA-sensitized guinea pigs increased significantly by 30 min after an acute exposure to 0.3% OVA. In addition, levels of eotaxin mRNA also increased in these tissues, but the increases were not significant until 2 h after challenge. Correspondingly, the number of eosinophils in bronchoalveolar lavage fluid did not increase until 4 h postchallenge. Results of these studies suggest that the OVA-sensitized guinea pig responds to allergic challenge with enhanced expression of genes (e.g., eotaxin, Muc2, and Muc5ac) that likely play a role in increased airway inflammation and mucin overproduction, and enhanced mucin gene expression appears to occur before eosinophil infiltration.}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Li, YH and Martin, LD and Minnicozzi, M and Greenfeder, S and Fine, J and Pettersen, CA and Chorley, B and Adler, KB}, year={2001}, month={Nov}, pages={644–651} }
 @inbook{martin_macchione_bonner_booth_akley_adler_2001, title={Interleukin-13 induced mucous cell hyperplasia in airway epithelium.}, booktitle={Cilia and mucus: from development to respiratory disease.}, author={Martin, L. D. and Macchione, M. and Bonner, J. C. and Booth, B. W. and Akley, N. J. and Adler, K. B.}, year={2001}, pages={253–263} }
 @article{booth_adler_bonner_tournier_martin_2001, title={Interleukin-13 induces proliferation of human airway epithelial cells in vitro via a mechanism mediated by transforming growth factor-alpha}, volume={25}, ISSN={["1535-4989"]}, DOI={10.1165/ajrcmb.25.6.4659}, abstractNote={Remodeling of the airways, as occurs in asthmatic patients, is associated with the continual presence of inflammatory mediators and Th2 cytokines, especially interleukin (IL)-13, during cycles of epithelial injury and repair. In this study, we examined the effect of IL-13 on well-differentiated normal human bronchial epithelial (NHBE) cells maintained in air-liquid interface culture. IL-13 induced proliferation of NHBE cells after 24 h exposure, as reflected by [(3)H]thymidine uptake and cell counts. The effects of IL-13 were mediated through the epidermal growth factor receptor (EGFR), as proliferation was attenuated by AG1478, an EGFR tyrosine kinase inhibitor. Proliferation appeared to be mediated by transforming growth factor (TGF)-alpha, a potent ligand for EGFR, which was released rapidly from NHBE cells in response to IL-13. Neutralizing antibody to TGF-alpha, but not antibodies against other potentially important growth factors (EGF, heparin binding epidermal growth factor-like growth factor [HB-EGF], platelet-derived growth factor [PDGF]), inhibited the mitogenic response to IL-13. This study provides the first experimental evidence that IL-13 can initiate a proliferative response of human airway epithelium in the absence of inflammatory cells or other cell types. The results are consistent with a mechanism whereby IL-13 induces release of TGF-alpha from the epithelial cells, which in turn binds via an autocrine/paracrine-type action to the EGFR, initiating proliferation. IL-13-induced airway remodeling in vivo may involve this epithelium-driven response.}, number={6}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Booth, BW and Adler, KB and Bonner, JC and Tournier, F and Martin, LD}, year={2001}, month={Dec}, pages={739–743} }
 @article{booth_bonner_akley_macchione_adler_martin_2001, title={Interleukin-13, a mediator of subepithelial fibrosis, enhances growth factor production and proliferation in human airway epithelial cells}, volume={120}, ISSN={["0012-3692"]}, DOI={10.1378/chest.120.1_suppl.S15}, abstractNote={Subepithelial fibrosis is a prominent feature of the remodeled asthmatic airway. The cytokine interleukin (IL)-13, implicated as a mediator in the development of asthma, induces a significant degree of subepithelial fibrosis in the lungs of transgenic mice. Since IL-13 has been shown to exert effects on the airway epithelium, including the development of a mucous phenotype, we have begun to determine whether IL-13 provokes production of factors from the epithelium that could elicit the observed subepithelial fibrotic response. In the studies reported herein, injured airways with regions of regenerating/differentiating cells and regions of normal fully differentiated cells have been mimicked by examining the effects of IL-13 on normal human bronchial epithelial cells during mucociliary differentiation in air/liquid interface culture. Exposure of normal human bronchial epithelial cells to IL-13 resulted in increased production of soluble transforming growth factor (TGF)-a, with the growth factor interacting in an autocrine manner with the epidermal growth factor receptor. Production of soluble TGF-a was very rapid, with a threefold increase observed in response to IL-13 (10 ng/mL) b y1ho fexposure. Continuous exposure to IL-13 throughout the course of mucociliary differentiation (a total of 10 days) resulted in a twofold increase in cell number by day 7 when cells are differentiated. Exposure to IL-13 (10 ng/mL; 24 h) provoked a threefold increase in proliferation once the cells were differentiated, an effect that could be duplicated in differ}, number={1}, journal={CHEST}, author={Booth, B and Bonner, J and Akley, N and Macchione, M and Adler, K and Martin, LD}, year={2001}, month={Jul}, pages={15S–15S} }
 @article{li_pettersen_martin_adler_2001, title={MARCKS protein interaction with the cellular contractile machinery may regulate mucin secretion by human airway epithelium.}, volume={163}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Li, Y. and Pettersen, C. A. and Martin, L. D. and Adler, K. B.}, year={2001}, pages={A225} }
 @article{li_martin_spizz_adler_2001, title={MARCKS protein is a key molecule regulating mucin secretion by human airway epithelial cells in vitro}, volume={276}, ISSN={["0021-9258"]}, DOI={10.1074/jbc.M105614200}, abstractNote={Hypersecretion of airway mucin characterizes numerous respiratory diseases. Although diverse pathological stimuli can provoke exocytotic release of mucin from secretory cells of the airway epithelium, mechanisms involved remain obscure. This report describes a new paradigm for the intracellular signaling mechanism regulating airway mucin secretion. Direct evidence is provided that the myristoylated alanine-rich C kinase substrate (MARCKS) is a central regulatory molecule linking secretagogue stimulation at the cell surface to mucin granule release by differentiated normal human bronchial epithelial cells in vitro. Down-regulation of MARCKS expression or disruption of MARCKS function in these cells inhibits the secretory response to subsequent stimulation. The intracellular mechanism controlling this secretory process involves cooperative action of two separate protein kinases, protein kinase C and cGMP-dependent protein kinase. Upon stimulation, activated protein kinase C phosphorylates MARCKS, causing translocation of MARCKS from the plasma membrane to the cytoplasm, where it is then dephosphorylated by a protein phosphatase 2A that is activated by cGMP-dependent protein kinase, and associates with both actin and myosin. Dephosphorylated cytoplasmic MARCKS would also be free to interact with mucin granule membranes and thus could link granules to the contractile cytoskeleton, mediating their movement to the cell periphery and subsequent exocytosis. These findings suggest several novel intracellular targets for pharmacological intervention in disorders involving aberrant secretion of respiratory mucin and may relate to other lesions involving exocytosis of membrane-bound granules in various cells and tissues.}, number={44}, journal={JOURNAL OF BIOLOGICAL CHEMISTRY}, author={Li, YH and Martin, LD and Spizz, G and Adler, KB}, year={2001}, month={Nov}, pages={40982–40990} }
 @inbook{li_martin_adler_2001, title={MARCKS protein: a potential modulator of airway mucin secretion.}, booktitle={Cilia and mucus: from development to respiratory disease.}, author={Li, Y. and Martin, L. D. and Adler, K. B.}, year={2001}, pages={179–193} }
 @article{bonner_wang_zhang_rice_zhang_adler_choe_kagan_2001, title={Role of receptor tyrosine kinases and mitogen-activated protein kinases in metal-induced pulmonary fibrosis}, volume={120}, ISSN={["0012-3692"]}, DOI={10.1378/chest.120.1_suppl.S55}, abstractNote={The proliferation of lung fibroblasts is a key component of pulmonary fibrosis. Several cell-surface receptor tyrosine kinases, including the platelet-derived growth factor receptor (PDGF-R) and epidermal growth factor receptor (EGF-R), mediate fibroblast mitogenesis via the activation of mitogen-activated protein (MAP) kinases. We have developed a model of metal-induced oxidative stress in rats using vanadium pentoxide (V2O5) that is characterized by interstitial and peribronchiolar fibrosis, airway smooth-muscle thickening, and mucous cell metaplasia. In vivo activation of the extracellular signal-regulated kinases (ERKs [ERK-1 and ERK-2]) was demonstrated by immunohistochemistry in fibrotic lesions caused by V2O5 exposure. Moreover, V2O5 injury upregulated platelet-derived growth factor α-receptor messenger RNA (mRNA) and protein in vivo. The mechanism of PDGF-Rα upregulation by V2O5 was elucidated in vitro and involved the release of interleukin-1β by alveolar macrophages, which then activated lung fibroblasts in a paracrine manner to activate p38 MAP kinase, which caused stabilization of PDGF-Rα mRNA. V2O5 also activated ERK-1 and ERK-2 in cultured lung fibroblasts in an oxidant-dependent manner that involved upstream activation of the EGF-R, Raf-1, MAP kinase kinase signaling cascade. In another study, V2O5 exposure of human bronchial epithelial cells in vitro caused the release of mitogenic activity for human lung fibroblasts that was abolished by a neutralizing antibody against heparin-binding epidermal growth factor-like growth factor. Induction of heparin-binding epidermal growth factor-like growth factor mRNA and protein by V2O5in vitro was reduced by the MAP kinase kinase inhibitor PD98059 and the p38 MAP kinase inhibitor SB203580. Finally, the intraperitoneal administration of tyrosine kinase inhibitors specific for either the PDGF-R or the EGF-R (tyrphostins AG1296 and AG1478, respectively) significantly reduced pulmonary fibrosis in rats exposed to V2O5. Collectively, these studies have identified signaling pathways and inducible genes activated by V2O5-stimulated oxidative stress that may offer potential targets for therapeutic intervention of pulmonary fibrosis.}, number={1}, journal={CHEST}, author={Bonner, JC and Wang, YZ and Zhang, P and Rice, A and Zhang, LM and Adler, K and Choe, N and Kagan, E}, year={2001}, month={Jul}, pages={55S–56S} }
 @article{zhang_rice_adler_sannes_martin_gladwell_koo_gray_bonner_2001, title={Vanadium stimulates human bronchial epithelial cells to produce heparin-binding epidermal growth factor-like growth factor - A mitogen for lung fibroblasts}, volume={24}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.24.2.4096}, abstractNote={The bronchial epithelium is a potential source of growth factors that could mediate airway fibrosis during the progression of diseases such as asthma and chronic bronchitis. We report that conditioned medium (CM) from normal human bronchial epithelial cells (NHBECs) contains mitogenic activity for human lung fibroblasts that is blocked by the epidermal growth factor receptor (EGF-R) tyrosine kinase inhibitor AG1478 and by neutralizing antibodies raised against heparin-binding epidermal growth factor-like growth factor (HB-EGF). Neutralizing antibodies against other EGF-R ligands (EGF and transforming growth factor-alpha) or other antibodies against growth factors (platelet-derived growth factors, insulin-like growth factor-1) had no affect on the mitogenic activity of NHBEC-CM. HB-EGF messenger RNA (mRNA) expression in NHBEC was detected by reverse transcriptase/polymerase chain reaction and Northern blot analysis. HB-EGF protein was detected by enzyme-linked immunosorbent assay. Vanadium pentoxide (V2O5), a fibrogenic metal associated with occupational asthma, caused a several-fold increase in HB-EGF mRNA expression and protein, whereas the inert metal titanium dioxide had no effect on HB-EGF expression. V2O5-induced HB-EGF mRNA expression was inhibited by the EGF-R tyrosine kinase inhibitor AG1478, the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580, and the MAP kinase kinase inhibitor PD98059. Finally, HB-EGF induced the production of fibroblast growth factor (FGF)-2 by human lung fibroblasts and anti-FGF-2 antibody partially blocked the mitogenic activity of NHBEC-CM on fibroblasts. These data suggest that HB-EGF is a fibroblast mitogen produced by NHBECs and that induction of an FGF-2 autocrine loop in fibroblasts by HB-EGF accounts for part of this mitogenic activity.}, number={2}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Zhang, LM and Rice, AB and Adler, K and Sannes, P and Martin, L and Gladwell, W and Koo, JS and Gray, TE and Bonner, JC}, year={2001}, month={Feb}, pages={123–131} }
 @inbook{he_adler_akley_fischer_jiang_krunkosky_martin_2000, title={Air-liquid interface culture systems for exposure of differentiated cells to oxidant stress.}, booktitle={Models and methods in cell signaling and gene expression applications to oxidative stress research.}, author={He, F. and Adler, K. B. and Akley, N. J. and Fischer, B. M. and Jiang, N. and Krunkosky, T. M. and Martin, L. D.}, year={2000}, pages={15–30} }
 @article{martin_krunkosky_adler_2000, title={Differential generation of intracellular oxidants by TNF? and exogenous superoxide affords distinct expression of IL-6 in human airway epithelium in vitro.}, volume={161}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Martin, L. D. and Krunkosky, T. M. and Adler, K. B.}, year={2000}, pages={A152} }
 @article{krunkosky_fischer_martin_jones_akley_adler_2000, title={Effects of TNF-alpha on expression of ICAM-1 in human airway epithelial cells in vitro - Signaling pathways controlling surface and gene expression}, volume={22}, ISSN={["1535-4989"]}, DOI={10.1165/ajrcmb.22.6.3925}, abstractNote={Signaling pathways associated with tumor necrosis factor (TNF)- α –induced intercellular adhesion molecule 1 (ICAM-1) surface and gene expression were investigated in well differentiated normal human bronchial epithelial (NHBE) cells in air–liquid interface primary culture. Cells were exposed to human recombinant TNF- α (hrTNF- α ; 0.015 to 150 ng/ml [specific activity, 2.86 × 107 U/mg]). TNF- α enhanced ICAM-1 surface expression (measured by flow cytometry) and steady-state messenger RNA (mRNA) levels (assessed by Northern hybridization) in concentration- and time-dependent manners. TNF- α –induced ICAM-1 surface and gene expression were both blocked by the RNA polymerase II inhibitor actinomycin D (0.1 μ g/ml), and surface expression was attenuated by a neutralizing monoclonal antibody directed against the TNF- α receptor p55 (TNF-RI). The intracellular signaling pathway leading to enhanced expression appeared to involve activation of a phospholipase C that hydrolyzes phosphatidylcholine (PC-PLC) becaus...}, number={6}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Krunkosky, TM and Fischer, BM and Martin, LD and Jones, N and Akley, NJ and Adler, KB}, year={2000}, month={Jun}, pages={685–692} }
 @article{bonner_zhang_sannes_martin_gladwell_koo_gray_adler_2000, title={Induction of heparin-binding epidermal growth factor (HB-EGF) mRNA in normal human bronchial epithelial cells by metal-induced oxidative stress.}, volume={161}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Bonner, J. C. and Zhang, L. and Sannes, P. and Martin, L. and Gladwell, W. and Koo, J.-S. and Gray, T. and Adler, K.}, year={2000}, pages={A149} }
 @article{martin_bonner_macchione_booth_akley_adler_2000, title={Interaction of TGF? and EGF receptor mediates IL-13 induced mucous cell hyperplasia in human airway epithelium in vitro.}, volume={161}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Martin, L. D. and Bonner, J. C. and Macchione, M. and Booth, B. and Akley, N. and Adler, K. B.}, year={2000}, pages={A779} }
 @article{li_martin_adler_2000, title={MARCKS protein: a key intracellular molecule controlling mucin secretion by human airway goblet cells.}, volume={161}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Li, Y. and Martin, L. D. and Adler, K. B.}, year={2000}, pages={A259} }
 @article{he_martin_adler_2000, title={Mechanisms of human neutrophil elastase-induced mucin hypersecretion by human airway epithelial cells in vitro.}, volume={161}, journal={American Journal of Respiratory and Critical Care Medicine}, author={He, F. and Martin, L. D. and Adler, K. B.}, year={2000}, pages={A155} }
 @article{adler_li_martin_2000, title={Myristoylated alanine-rich C-kinase substrate protein: A major intracellular regulatory molecule controlling secretion of mucin by human airway goblet cells.}, volume={117}, number={5 Supplement 1}, journal={Chest}, author={Adler, K. B. and Li, Y. and Martin, L. D.}, year={2000}, pages={266S–267S} }
 @article{jiang_dreher_dye_li_richards_martin_adler_2000, title={Residual oil fly ash induces cytotoxicity and mucin secretion by guinea pig tracheal epithelial cells via an oxidant-mediated mechanism}, volume={163}, ISSN={["0041-008X"]}, DOI={10.1006/taap.1999.8886}, abstractNote={Inhalation of ambient air particulate matter (PM) is associated with pulmonary injury and inflammation. Using primary cultures of guinea pig tracheal epithelial (GPTE) cells as an in vitro model of airway epithelium, we examined effects of exposure to suspensions of six different emission and ambient air PM samples: residual oil fly ash (ROFA) from an electrical power plant; fly ash from a domestic oil burning furnace (DOFA); ambient air dust from St. Louis (STL), Ottawa (OT), and Washington, DC (WDC); and volcanic ash from the eruption of Mount Saint Helens (MSH) in 1980. Effects of these particulates on cell viability (assessed via LDH assay), secretion of mucin (measured by a monoclonal antibody-based ELISA), and steady-state mRNA levels of the mucin gene MUC2 were determined. ROFA was the most toxic of the dusts tested, as it significantly increased LDH release following a 24-h incubation with 50 microg/cm(2) ROFA. ROFA also enhanced MUC2 mRNA after 4-h exposure, and mucin secretion after 8 h. ROFA-induced mucin secretion and cytotoxicity were attenuated by the oxidant scavenger, dimethylthiourea (DMTU). ROFA exposure also depleted cells of glutathione (GSH). Relatedly, depletion of intracellular GSH by treatment of the cells with buthionine sulfoxamine (BSO) also provoked mucin secretion, as well as enhancing the secretory effect of ROFA when the two agents were added together. L-NMA, the nitric oxide synthase (NOS) inhibitor, did not affect ROFA-induced mucin secretion. Of the soluble transition metals in ROFA (nickel, iron, vanadium), only vanadium individually, or combinations of the metals containing vanadium, provoked secretion. The results suggest ROFA enhances mucin secretion and generates toxicity in vitro to airway epithelium via a mechanism(s) involving generation of oxidant stress, perhaps related to depletion of cellular antioxidant capacity. Deleterious effects of inhalation of ROFA in the respiratory tract in vivo may relate to these cellular responses. Vanadium, a component of ROFA, may be important in generating these reactions.}, number={3}, journal={TOXICOLOGY AND APPLIED PHARMACOLOGY}, author={Jiang, NF and Dreher, KL and Dye, JA and Li, YH and Richards, JH and Martin, LD and Adler, KB}, year={2000}, month={Mar}, pages={221–230} }
 @article{martin_norford_voynow_adler_2000, title={Response of human airway epithelium in vitro to inflammatory mediators - Dependence on the state of cellular differentiation}, volume={117}, ISSN={["0012-3692"]}, DOI={10.1378/chest.117.5_suppl_1.267s}, abstractNote={A irways in patients with COPD or chronic bronchitis contain regions of damaged and regenerating epithelium intermixed with normally differentiated mucociliary areas. Responses of these different regions to the inflammatory milieu present in airways of these individuals may differ, thereby altering further development of additional lesions in the airways. In the studies reported here, we utilized normal human bronchial epithelial (NHBE) cells cultured in an air/liquid interface system as a model of well-differentiated epithelium, and the same cells cultured on plastic and submerged in medium as a model of poorly differentiated, regenerating epithelium. We investigated the responses of these different cell types to inflammatory mediators present in inflamed airways: the cytokine interleukin (IL)-13, human neutrophil elastase (HNE), and “cytomix” (10 ng/mL tumor necrosis factor-a, interferon-g, and IL-1b). Acute exposure to IL-13 (10 ng/mL, 24 h) caused an increase in steady-state messenger RNA (mRNA) for mucin (MUC5AC) in undifferentiated cells, but did not affect MUC5AC expression in differentiated cells. Secretion of mucin and the secondary cytokine, IL-6, were both decreased in differentiated epithelial cell cultures after exposure to IL-13, but no secretory change was observed in undifferentiated cells. By contrast, chronic exposure to IL-13 (10 ng/mL, 8 days) caused an increase in mucin secretion in differentiated airway epithelial cells, and a decrease in undifferentiated cells. In response to HNE, well-differentiated cells increased steady-state levels of MUC5AC mRNA, but undifferentiated cells increased mRNA levels of another mucin gene, MUC4. Finally, the message level of inducible nitric oxide synthase (NOS) was increased by cytomix only in differentiated NHBE cultures. Undifferentiated cells did not express inducible NOS at all, but rather the constitutive forms of NOS, endothelial NOS and brain NOS. These data suggest that the response of the airway epithelium to inflammatory mediators may be markedly different in undifferentiated vs fully differentiated cells, and these responses may play a role in further exacerbation of airway inflammation. In vitro studies utilizing cultured airway epithelial cells must take the state of differentiation of these cells into account when analyzing such responses and extrapolating to the in vivo situation.}, number={5}, journal={CHEST}, author={Martin, LD and Norford, D and Voynow, J and Adler, KB}, year={2000}, month={May}, pages={267S–267S} }
 @article{li_greenfeder_martin_minnicozzi_voynow_adler_1999, title={Cloning and expression of guinea pig MUC2 and MUC5AC genes.}, volume={159}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Li, Y. and Greenfeder, S. and Martin, L. D. and Minnicozzi, M. and Voynow, J. A. and Adler, K. B.}, year={1999}, pages={A852} }
 @article{li_joshee_adler_cheng_1999, title={Development of monoclonal antibodies against bovine mucin core 2 beta 6 N-acetylglucosaminyltransferase}, volume={16}, ISSN={["1573-4986"]}, DOI={10.1023/A:1007030223118}, abstractNote={Molecular cloning techniques have been used to produce abundant amounts of recombinant glycosyltransferases for biochemical studies. We recently cloned a cDNA which encoded bovine mucin core 2 beta6N-acetylglucosaminyl transferase (C2TF). Poly-histidine-C2TF fusion protein was generated from the cloned cDNA in the E. coli Xpress system and used to produce monoclonal antibodies (MAbs). We obtained seven hybridomas which secreted MAbs against bovine C2TF in mouse ascites with titers ranging from 1:1280 to 1:40960 as assessed by immunofluorescence assay (IF). Isotyping revealed that all seven MAbs were IgG (4 IgG1, 2 IgG2b and 1 IgG2a). The affinity constants (M(-1)) for these MAbs range from 5.4 x 10(7) to 1.2 x 10(9). These MAbs recognized bovine C2TF in tissue sections and on Western blottings. Six of these MAbs reacted with human core 2-M enzyme and one with both core 2-L and core 2-M enzymes on Western blottings. Therefore, these antibodies should be useful for further study of bovine and human core 2 enzymes.}, number={9}, journal={GLYCOCONJUGATE JOURNAL}, author={Li, CM and Joshee, N and Adler, KB and Cheng, PW}, year={1999}, month={Sep}, pages={555–562} }
 @article{li_he_martin_krunkosky_lincoln_cornwell_adler_1999, title={Myristoylated alanine-rich C kinase substrate (MARCKS) is produced by human airway epithelial cells and is phosphorylated by PKC and PKG.}, volume={159}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Li, Y. and He, F. and Martin, L. D. and Krunkosky, T. M. and Lincoln, T. M. and Cornwell, T. L. and Adler, K. B.}, year={1999}, pages={A723} }
 @article{zhang_adler_sannes_bonner_1999, title={Normal human bronchial epithelial cells produce heparin-binding epidermal growth factor (HB-EGF), a mitogen for human lung fibroblasts in vitro.}, volume={159}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Zhang, L. and Adler, K. B. and Sannes, P. and Bonner, J. C.}, year={1999}, pages={A508} }
 @article{dye_adler_richards_dreher_1999, title={Role of soluble metals in oil fly ash-induced airway epithelial injury and cytokine gene expression}, volume={277}, ISSN={["1040-0605"]}, DOI={10.1152/ajplung.1999.277.3.l498}, abstractNote={Particulate matter (PM) metal content and bioavailability have been hypothesized to play a role in the health effects epidemiologically associated with PM exposure, in particular that associated with emission source PM. Using rat tracheal epithelial cells in primary culture, the present study compared and contrasted the acute airway epithelial effects of an emission source particle, residual oil fly ash (ROFA), with that of its principal constitutive transition metals, namely iron, nickel, and vanadium. Over a 24-h period, exposure to ROFA, vanadium, or nickel plus vanadium, but not to iron or nickel, resulted in increased epithelial permeability, decreased cellular glutathione, cell detachment, and lytic cell injury. Treatment of vanadium-exposed cells with buthionine sulfoximine further increased cytotoxicity. Conversely, treatment with the radical scavenger dimethylthiourea inhibited the effects in a dose-dependent manner. RT-PCR analysis of RNA isolated from ROFA-exposed rat tracheal epithelial cells demonstrated significant macrophage inflammatory protein-2 and interleukin-6 gene expression as early as 6 h after exposure, whereas gene expression of inducible nitric oxide synthase was maximally increased 24 h postexposure. Again, vanadium (not nickel) appeared to be mediating the effects of ROFA on gene expression. Treatment with dimethylthiourea inhibited both ROFA- and vanadium-induced gene expression in a dose-dependent manner. Corresponding effects were observed in interleukin-6 and macrophage inflammatory protein-2 synthesis. In summary, generation of an oxidative stress was critical to induction of the ROFA- or vanadium-induced effects on airway epithelial gene expression, cytokine production, and cytotoxicity.}, number={3}, journal={AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY}, author={Dye, JA and Adler, KB and Richards, JH and Dreher, KL}, year={1999}, month={Sep}, pages={L498–L510} }
 @article{adler_martin_voynow_krunkosky_1999, title={TNF? effects on human airway epithelial cells in vitro: molecular mechanisms of enhanced mucin production/secretion and ICAM-I expression.}, volume={8}, journal={404nOtfound}, author={Adler, K. B. and Martin, L. D. and Voynow, J. A. and Krunkosky, T. M.}, year={1999}, pages={S55} }
 @article{krunkosky_martin_li_adler_1999, title={TNF?-induced ICAM-1 expression in airway epithelium: involvement of IkB?.}, volume={159}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Krunkosky, T. M. and Martin, L. D. and Li, Y. and Adler, K. B.}, year={1999}, pages={A184} }
 @article{martin_krunkosky_adler_1999, title={Transcription factor activation during oxidant-regulated interleukin-6 expression in normal human bronchial epithelium in vitro.}, volume={159}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Martin, L. D. and Krunkosky, T. M. and Adler, K. B.}, year={1999}, pages={A447} }
 @article{fischer_rochelle_voynow_akley_adler_1999, title={Tumor necrosis factor-alpha stimulates mucin secretion and cyclic GMP production by guinea pig tracheal epithelial cells in vitro}, volume={20}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.20.3.3393}, abstractNote={Tumor necrosis factor (TNF)-alpha, a pluripotent cytokine implicated in the pathogenesis of airway inflammation, has been shown to provoke hypersecretion of mucin by airway epithelial cells in vitro. In this study, we investigated potential signaling pathways mediating TNF-alpha-induced mucin secretion using guinea pig tracheal epithelial (GPTE) cells in air-liquid interface culture. Exogenously applied TNF-alpha (human recombinant) stimulated mucin secretion in a concentration-dependent manner, with maximal effects at 10 to 15 ng/ml (286 to 429 U/ml). The pathway of stimulated secretion appeared to involve generation of intracellular nitric oxide (NO), activation of soluble guanylate cyclase (GC-S), production of cyclic guanosine monophosphate (cGMP), and activation of cGMP-dependent protein kinase (PKG). TNF-alpha increased production of nitrite and nitrate by GPTE cells; both mucin secretion and cGMP production were attenuated by NG-monomethyl-L-arginine (1 mM), a competitive inhibitor of nitric oxide synthase (NOS), or by the GC-S inhibitor LY83583 (50 microM); and mucin secretion in response to TNF-alpha or to the cGMP analogue dibutyryl cGMP (100 and 500 microM) was attenuated by the specific PKG inhibitor KT5823 (1 microM). Increased mucin secretion and increased cGMP production in response to TNF-alpha both appeared to be mediated by a phospholipase C that hydrolyzes phosphatidylcholine (PC-PLC), and by protein kinase C (PKC), since both responses were attenuated by either D609 (10 and 20 microg/ml), a specific PC-PLC inhibitor, or by each of three PKC inhibitors: Calphostin C (0.3 and 0.5 microM), bisindoylmaleimide (GF 109203X, Go 6850; 20 nM), or Ro31-8220 (10 microM). Collectively, the results suggest that TNF-alpha stimulates secretion of mucin by GPTE cells via a mechanism(s) dependent on PC-PLC and PKC, and involving activation of NOS, generation of NO, production of cGMP, and activation of PKG.}, number={3}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Fischer, BM and Rochelle, LG and Voynow, JA and Akley, NJ and Adler, KB}, year={1999}, month={Mar}, pages={413–422} }
 @article{kim_koo_norford_gray_adler_nettesheim_1999, title={Upregulation of mucin gene expression in airway epithelial cells by inflammatory mediators.}, volume={159}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Kim, Y. S. and Koo, J. S. and Norford, D. and Gray, T. and Adler, K. and Nettesheim, P.}, year={1999}, pages={A36} }
 @article{li_martin_minnicozzi_adler_1998, title={Cloning of guinea pig Muc2 cDNA and MUC2 gene expression in guinea pig airway epithelium in vitro.}, volume={157}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Li, Y. and Martin, L. D. and Minnicozzi, M. and Adler, K. B.}, year={1998}, pages={A728} }
 @article{rochelle_fischer_adler_1998, title={Concurrent production of reactive oxygen and nitrogen species by airway epithelial cells in vitro}, volume={24}, ISSN={["0891-5849"]}, DOI={10.1016/S0891-5849(97)00375-4}, abstractNote={Intracellularly generated reactive species of both oxygen (ROS) and nitrogen (RNS) have been implicated in signaling responses in airway epithelial cells, but these radicals have not been measured directly in such cells. In this study, intracellular production of both ROS and RNS were measured in the same cell lysates of guinea pig tracheal epithelial (GPTE) cells maintained in primary culture. ROS and RNS were quantified under basal (constitutive) conditions and in response to different stimuli: LPS and TNFα [activators of inducible nitric oxide synthase (iNOS)]; several activators of calcium-dependent cNOS (ATP, bradykinin, ionophore A23187, and thapsigargin); and exogenous oxidant stress generated by addition of xanthine oxidase to purine (p + XO). Studies with LPS and TNFα also were performed using the murine macrophage cell line, RAW 264.7, as a positive control. Intracellular oxidant production was detected from oxidation of dihydrorhodamine to rhodamine. NOx was quantified by either chemiluminescent or fluorescent detection. NOS activity was measured as citrulline production from arginine. Basal production of oxidants by GPTE cells (0.08 + 0.00 nmol rhodamine) was less than 10% that of RAW.267 cells (0.91 + 0.03 nmol rhodamine). TNFα and LPS significantly increased intracellular oxidant production in GPTE cells, as did p + XO, but none of the cNOS activators affected production of oxidants in these cells. Concentrations of NO2 after 4 h in unstimulated RAW 264.7 and GPTE cells were similar and comprised 63% of total NOx in GPTE and 62% in RAW cells. TNFα and LPS both increased NO2 in GPTE cells, but none of the Ca++-mobilizing agents nor p + XO significantly affected intracellular RNS. The results suggest both ROS and RNS can be measured in the same lysates from airway epithelial cells, and that both ROS and RNS are produced in these cells in response to different stimuli.}, number={5}, journal={FREE RADICAL BIOLOGY AND MEDICINE}, author={Rochelle, LG and Fischer, BM and Adler, KB}, year={1998}, month={Mar}, pages={863–868} }
 @article{adler_fischer_martin_voynow_1998, title={Effects of inflammatory mediators and drugs on mucus secretion and mucociliary function.}, volume={149}, journal={404nOtfound}, author={Adler, K. B. and Fischer, B. M. and Martin, L. D. and Voynow, J. A.}, year={1998}, pages={245–248} }
 @article{adler_jiang_dye_dreher_1998, title={Exposure of differentiated rodent airway epithelial cells in vitro to particles of residual fly ash (ROFA) induces cytotoxicity and generation of reactive oxygen species.}, volume={10}, journal={Proceedings of the 10th International Colloquium on Lung Fibrosis}, author={Adler, K. B. and Jiang, N. F. and Dye, J. A. and Dreher, K. L.}, year={1998}, pages={26} }
 @article{norford_koo_gray_alder_nettesheim_1998, title={Expression of nitric oxide synthase isoforms in normal human tracheobronchial epithelial cells in vitro: Dependence on retinoic acid and the state of differentiation}, volume={24}, ISSN={["0190-2148"]}, DOI={10.3109/01902149809041540}, abstractNote={The retinoic acid (RA) and differentiation dependence of constitutive expression of the nitric oxide synthase (NOS) isoforms, iNOS, eNOS, and bNOS, was examined by reverse transcriptase polymerase chain recitation (RT-PCR) in cultured, normal, human, tracheobronchial epithelial (NHTBE) cells. In the presence of RA (RA+), early passage NHTBE cells grown in air-liquid interface (ALI) cultures undergo mucous differentiation; in the absence of RA (RA-), they undergo metaplastic squamous differentiation. Under both conditions the respective differentiated phenotype develops around day 10 of culture. We found that iNOS mRNA levels were much higher in RA+ cultures, expressing the mucous phenotype, than in RA- cultures, expressing the metaplastic squamous phenotype. In contrast, eNOS mRNA levels were much higher in RA- cultures than in RA+ cultures. Expression of bNOS was not significantly affected by the RA status. The pattern of expression of NOS isoforms was then studied during the course of development of the two cellular phenotypes. During the early stages of differentiation, expression of iNOS (RA+) and eNOS (RA-) was very low, indicating that the expression of these two isoforms was not only dependent on the presence or absence of RA, but also on the degree of differentiation. The differentiation dependence of bNOS mRNA was less obvious. Four days of RA treatment of RA- cultures, which reverses the squamous phenotype and restores mucous differentiation, induced iNOS expression in a concentration-dependent manner. eNOS expression was depressed by 10(-8) M RA, while bNOS mRNA levels were slightly reduced by 10(-6) M RA. No NOS proteins were detected in unstimulated RA+ and RA- cultures. iNOS protein was induced by cytokine treatment in RA+ cultures, in contrast to eNOS and bNOS protein levels, which were unaffected. Our studies show that constitutive expression of the NOS isoforms is differentially regulated and that iNOS and eNOS mRNA levels are dependent on the stage of mucous and squamous differentiation, respectively. bNOS expression was only marginally affected by the RA or differentiation status.}, number={3}, journal={EXPERIMENTAL LUNG RESEARCH}, author={Norford, D and Koo, JS and Gray, T and Alder, K and Nettesheim, P}, year={1998}, pages={355–366} }
 @article{martin_krunkosky_voynow_adler_1998, title={Intracellular reactive oxygen and nitrogen species as signaling molecules in airway epithelium.}, volume={106}, journal={Environmental Health Perspectives}, author={Martin, L. D. and Krunkosky, T. M. and Voynow, J. A. and Adler, K. B.}, year={1998}, pages={1197–1203} }
 @inbook{fischer_li_li_choe_wright_rochelle_adler_1998, title={Intracellular regulation of airway mucin secretion.}, booktitle={Cilia, mucus and mucociliary interactions}, author={Fischer, B. M. and Li, C. and Li, H. and Choe, N. and Wright, D. T. and Rochelle, L. G. and Adler, K. B.}, year={1998}, pages={143–151} }
 @article{krunkosky_fischer_dodam_adler_1998, title={Mechanisms of TNF? signaling in airway epithelial cells in vitro.}, volume={157}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Krunkosky, T. M. and Fischer, B. M. and Dodam, J. R. and Adler, K. B.}, year={1998}, pages={A198} }
 @article{li_adler_cheng_1998, title={Mucin biosynthesis: Molecular cloning and expression of bovine lung mucin core 2 N-acetylglucosaminyltransferase cDNA}, volume={18}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.18.3.2593}, abstractNote={A cDNA clone containing a 2,150-bp insert was isolated from a bovine lung lambdagt10 cDNA library by cross-species hybridization using a DNA probe generated by polymerase chain reaction (PCR) employing a human cDNA that encodes mucin core 2 beta6-N-acetylglucosaminyltransferase (hC2TF) as the template. The bovine cDNA (bcDNA) insert was devoid of 220 bp of the 5' portion of the C2TF open reading frame (ORF), as predicted from the human counterpart. Southern blotting analysis suggested that the coding region of this C2TF gene is in one exon. To construct a full-length bovine C2TF (bC2TF) cDNA, a genomic DNA fragment containing the 5' portion of the ORF of the bC2TF gene was cloned from a lambdaEMBL bovine genomic DNA library and ligated to the 5' end of the cloned cDNA insert. DNA sequence analysis showed that the complete ORF of bC2TF gene was 1,281 bp in length, which corresponds to a polypeptide of 427 amino acids. Catalytically active bC2TF was expressed in sf21 insect cells infected with recombinant baculovirus containing the ORF of the bC2TF gene. The recombinant bC2TF catalyzed the synthesis of core 2, but not core 4 and blood group I structures. Western blotting analysis showed that the recombinant bC2TF migrated with the same mobility (approximately 55 kD) as the native bovine tracheal C2TF. Immunohistochemical analysis showed that in bovine trachea, the bC2TF was present at the surface epithelium and in the submucosal glands, with the latter being the major site of distribution.}, number={3}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Li, CM and Adler, KB and Cheng, PW}, year={1998}, month={Mar}, pages={343–352} }
 @inbook{martin_krunkosky_voynow_adler_1998, title={Oxidant-regulated gene expression in inflammatory lung disease.}, DOI={10.1007/978-1-4419-8634-4_23}, booktitle={Acute respiratory distress syndrome: cellular and molecular mechanisms and clinical management.}, author={Martin, L. D. and Krunkosky, T. M. and Voynow, J. A. and Adler, K. B.}, year={1998}, pages={187–195} }
 @article{adler_jiang_dye_dreher_1998, title={Particles of Residual Oil Fly Ash (ROFA) induce toxicity and mucin hypersecretion in rodent airway epithelial cells in vitro via an oxidant-mediated mechanism.}, volume={95}, DOI={10.1016/s0378-4274(98)80888-6}, journal={Toxicology Letters}, author={Adler, K. B. and Jiang, N. F. and Dye, J. A. and Dreher, K. L.}, year={1998}, pages={224} }
 @article{norford_koo_gray_adler_1998, title={Regulation of nitric oxide synthase (NOS) isoforms in human tracheobronchial cells by retinoic acid.}, volume={157}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Norford, D. and Koo, J. S. and Gray, T. and Adler, K. B.}, year={1998}, pages={A769} }
 @article{jiang_dreher_adler_1998, title={Residual oil fly ash (ROFA) induces cytotoxicity and enhances mucin secretion by guinea pig airway epithelial cells in vitro via an oxidant mediated mechanism.}, volume={157}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Jiang, N.-F. and Dreher, K. L. and Adler, K. B.}, year={1998}, pages={A150} }
 @article{martin_krunkosky_voynow_adler_1998, title={The role of reactive oxygen and nitrogen species in airway epithelial gene expression}, volume={106}, ISSN={["0091-6765"]}, DOI={10.2307/3433986}, abstractNote={The body first encounters deleterious inhaled substances, such as allergens, industrial particles, pollutants, and infectious agents, at the airway epithelium.When this occurs, the epithelium and its resident inflammatory cells respond defensively by increasing production of cytokines, mucus, and reactive oxygen and nitrogen species (ROS/RNS).As inflammation in the airway increases, additional infiltrating cells increase the level of these products.Recent interest has focused on ROS/RNS as potential modulators of the expression of inflammation-associated genes important to the pathogenesis of various respiratory diseases.ROS/RNS appear to play a variety of roles that lead to changes in expression of genes such as interleukin-6 and intercellular adhesion molecule 1.By controlling this regulation, the reactive species can serve as exogenous stimuli, as intercellular signaling molecules, and as modulators of the redox state in epithelial cells.Unraveling the molecular mechanisms affected by ROS/RNS acting in these capacities should aid in the understanding of how stimulated defense mechanisms within the airway can lead to disease.}, journal={ENVIRONMENTAL HEALTH PERSPECTIVES}, author={Martin, LD and Krunkosky, TM and Voynow, JA and Adler, KB}, year={1998}, month={Oct}, pages={1197–1203} }
 @article{dye_adler_rochelle_dreher_1998, title={Vanadium content and related oxidative stress appear to determine airway epithelial cell responses to emission source particulate matter.}, volume={12}, journal={FASEB Journal}, author={Dye, J. A. and Adler, K. B. and Rochelle, L. G. and Dreher, K. L.}, year={1998}, pages={A337} }
 @article{dye_adler_richards_dreher_1997, title={Airway epithelial cell responses to fly ash (ROFA) particles: contribution of soluble transition metals.}, volume={155}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Dye, J. A. and Adler, K. B. and Richards, J. H. and Dreher, K. L.}, year={1997}, pages={A197} }
 @inbook{cohn_fischer_adler_1997, title={Airway epithelial cells in asthma.}, booktitle={Allergy and allergic diseases.}, author={Cohn, L. A. and Fischer, B. M. and Adler, K. B.}, year={1997}, pages={263–283} }
 @misc{martin_rochelle_fischer_krunkosky_adler_1997, title={Airway epithelium as an effector of inflammation: molecular regulation of secondary mediators}, volume={10}, ISSN={["0903-1936"]}, DOI={10.1183/09031936.97.10092139}, abstractNote={Deleterious environmental stimuli cause the airway epithelium to respond with increased secretions of mucus, reaction of oxygen/nitrogen species, changes in ciliary beating, and the influx of inflammatory cells. The epithelium is a target for factors released by infiltrating inflammatory cells, and has recently been shown to serve as an effector of such inflammation. Molecular mechanisms regulating production of secondary inflammatory mediators (cytokines, lipid mediators, and reactive oxygen/nitrogen species) have yet to be fully described. This report reviews the production of secondary mediators by epithelial cells and by airway epithelium. Lipid mediators are enzymatically produced by the airway epithelium in response to primary mediators. Molecular mechanisms regulating the production of cyclo-oxygenase, lipoxygenase and prostaglandin synthase are discussed, along with the potential of lipid mediators to produce inflammation. The molecular regulation of nitric oxide production is also described in the context of its role as a signalling molecule in pathways regulating secretion of mucus, ciliary motion, and intercellular adhesion molecule-1 (ICAM-1) expression. The production of cytokines by the airway epithelium is shown to play a role in causing inflammation associated with respiratory diseases. Particular attention is paid to molecular mechanisms governing the expression of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and interleukin-8 (IL-8).}, number={9}, journal={EUROPEAN RESPIRATORY JOURNAL}, author={Martin, LD and Rochelle, LG and Fischer, BM and Krunkosky, TM and Adler, KB}, year={1997}, month={Sep}, pages={2139–2146} }
 @article{adler_krunkosky_fischer_1997, title={Effects of tumour necrosis factor-alpha on airway epithelial cells in vitro.}, volume={3}, journal={404nOtfound}, author={Adler, K. B. and Krunkosky, T. M. and Fischer, B. M.}, year={1997}, pages={74} }
 @article{dye_adler_richards_dreher_1997, title={Epithelial injury induced by exposure to residual oil fly-ash particles: Role of reactive oxygen species?}, volume={17}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.17.5.2749}, abstractNote={Exposure of animals to airborne particulates is associated with pulmonary injury and inflammation. In the studies described here, primary cultures of rat tracheal epithelial (RTE) cells were exposed to suspensions of residual oil fly ash (ROFA). ROFA exposure resulted in progressive cytotoxicity whereby the amount of lactate dehydrogenase (LDH) released was significantly greater at 24 h than at 6 h after exposure. In a dose-dependent manner, exposure to 5, 10, or 20 microg/cm2 of ROFA for 24 h resulted in cytotoxicity and detachment of cells from the collagen matrix, along with altered permeability of the RTE cell layer. ROFA exposure caused cellular glutathione levels to decrease, producing a condition of oxidative stress in the RTE cells. Treatment of RTE cells with buthionine sulfoxamine, an inhibitor of gamma-glutamyl cysteine synthetase, was found to augment ROFA-induced cytotoxicity. Treatment with dimethylthiourea (DMTU) inhibited ROFA-induced LDH release and permeability changes in a dose-dependent manner. Treatment with the nitric oxide synthase inhibitor NG-monomethyl-D-arginine (D-NMA) for 24 h was without effect. In rats intratracheally instilled with ROFA (500 microg/rat), intraperitoneal administration of DMTU (500 mg/kg) significantly ameliorated the degree of pulmonary neutrophilic inflammation present at 24 h. Overall, these in vitro findings suggest that ROFA-induced RTE cell injury may be mediated by hydroxyl-radical-like reactive oxygen species (i.e., species scavenged by DMTU) that are generated via non-nitric oxide pathways. The delay in induction of maximal RTE cell injury may reflect the time necessary to produce an oxidative burden by depleting antioxidant defenses such as cellular glutathione.}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={Dye, JA and Adler, KB and Richards, JH and Dreher, KL}, year={1997}, month={Nov}, pages={625–633} }
 @article{rochelle_fischer_boehm_akley_adler_1997, title={Inflammatory mediators increase intracellular reactive oxygen and nitrogen species (ROS/RNS) in airway epithelial cells.}, volume={155}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Rochelle, L. G. and Fischer, B. M. and Boehm, K. A. and Akley, N. J. and Adler, K. B.}, year={1997}, pages={A437} }
 @inbook{wright_li_fischer_yagaloff_adler_1997, title={Interactions between oxidant gases and airway epithelial cells; in vitro in vivo correlations.}, booktitle={Correlations between in vitro and in vivo investigations in inhalation toxicology.}, author={Wright, D. T. and Li, C. M. and Fischer, B. M. and Yagaloff, K. and Adler, K. B.}, year={1997}, pages={216–229} }
 @article{adler_li_li_choe_fischer_1997, title={Intracellular regulation of mucin secretion.}, journal={Proceedings of the International Congress on Cilia, Mucus and Mucociliary Interations.}, author={Adler, K. B. and Li, C. M. and Li, H. F. and Choe, N. and Fischer, B. M.}, year={1997}, pages={15} }
 @article{krunkosky_martin_fischer_adler_1997, title={Mechanisms of TNF?-induced ICAM-1 expression in airway epithelium.}, volume={155}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Krunkosky, T. M. and Martin, L. D. and Fischer, B. M. and Adler, K. B.}, year={1997}, pages={A207} }
 @article{cheng_li_adler_1997, title={Molecular cloning and expression of a mucin core 2 N-acetylglucosaminyltransferase cDNA.}, volume={155}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Cheng, P. W. and Li, C. M. and Adler, K. B.}, year={1997}, pages={A434} }
 @article{lee_madden_hatch_bottei_peden_adler_devlin_1997, title={Ozone induced DNA single strand breaks in human and guinea pig lung cells in vivo}, volume={9}, number={9}, journal={Inhalation Toxicology}, author={Lee, J. G. and Madden, M. C. and Hatch, G. and Bottei, G. and Peden, D. and Adler, K. B. and Devlin, R.}, year={1997}, pages={811–828} }
 @article{murphy_akley_chitapo_adler_1997, title={Secretory inhibition by airway epithelium increases with age.}, volume={155}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Murphy, T. M. and Akley, N. J. and Chitapo, P. and Adler, K.}, year={1997}, pages={A427} }
 @article{martin_krunkosky_rochelle_adler_1997, title={TNF? and oxidant-induced expression of Interleukin-6 in normal human bronchial epithelium in vitro.}, volume={155}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Martin, L. D. and Krunkosky, T. M. and Rochelle, L. G. and Adler, K. B.}, year={1997}, pages={A206} }
 @article{martin_krunkosky_dye_fischer_jiang_rochelle_akley_dreher_adler_1997, title={The role of reactive oxygen and nitrogen species in the response of airway epithelium to particulates}, volume={105}, ISSN={["1552-9924"]}, DOI={10.2307/3433551}, abstractNote={Epidemiologic and occupational studies indicate adverse health effects due to inhalation of particulate air pollutants, but precise biologic mechanisms responsible have yet to be fully established.The tracheobronchial epithelium forms the body's first physiologic barrier to such airborne pollutants, where ciliary movement functions to remove the offending substances caught in the overlying mucus layer.Resident and infiltrating phagocytic cells also function in this removal process.In this paper, we examine the role of reactive oxygen and nitrogen species (ROS/RNS) in the response of airway epithelium to particulates.Some particulates themselves can generate ROS, as can the epithelial cells, in response to appropriate stimulation.In addition, resident macrophages in the airways and the alveolar spaces can release ROS/RNS after phagocytosis of inhaled particles.These macrophages also release large amounts of tumor necrosis factor alpha (TNF-a), a cytokine that can generate responses within the airway epithelium dependent upon intracellular generation of ROS/RNS.As a result, signal transduction pathways are set in motion that may contribute to inflammation and other pathobiology in the airway.Such effects include increased expression of intercellular adhesion molecule 1, interleukin-6, cytosolic and inducible nitric oxide synthase, manganese superoxide dismutase, cytosolic phospholipase A2, and hypersecretion of mucus.Ultimately, ROS/RNS may play a role in the global response of the airway epithelium to particulate pollutants via activation of kinases and transcription factors common to many response genes.Thus, defense mechanisms involved in responding to offending particulates may result in a complex cascade of events that can contribute to airway pathology.}, journal={ENVIRONMENTAL HEALTH PERSPECTIVES}, author={Martin, LD and Krunkosky, TM and Dye, JA and Fischer, BM and Jiang, NF and Rochelle, LG and Akley, NJ and Dreher, KL and Adler, KB}, year={1997}, month={Sep}, pages={1301–1307} }
 @article{adler_menard_sannes_1996, title={Airway epithelium: A primary source for growth factors related to development of fibrosis?}, volume={9}, journal={Proceedings of the 9th International Colloquium on Lung Fibrosis}, author={Adler, K. B. and Menard, M. and Sannes, P. L.}, year={1996}, pages={29} }
 @article{dodam_adler_olson_1996, title={Effect of pertussis toxin and B-Oligomer on platelet-activating factor-induced generation of inositol phosphates in porcine alveolar macrophages.}, volume={57}, journal={American Journal of Veterinary Research}, author={Dodam, J. R. and Adler, K. B. and Olson, N. C.}, year={1996}, pages={574–579} }
 @article{rochelle_fischer_krunkosky_wright_adler_1996, title={Environmental toxins induce intracellular responses of airway epithelium through reactive species of oxygen and nitrogen}, volume={109}, ISSN={["0012-3692"]}, DOI={10.1378/chest.109.3_supplement.35s}, abstractNote={Pulmonary disorders and diseases, including asthma, ARDS, emphysema, and asbestosis have been linked to and exacerbated by inhalation of environmental pollutants such as ozone, oxides of nitrogen, and particulates. Airway epithelia are in a particularly vulnerable location with respect to these pollutants because they form the first physiologic barrier. In the tracheobronchial segment of the airways, ciliary movement and mucus secretion cooperate to remove offending substances. Mucus can provide a beneficial effect by sequestering exogenous pollutants and inactivating oxidant molecules, but excess secretion of mucus can lead to airway obstruction, increased microbial infection, and compromised exchange of ions and gases. Mucus secretion may be linked to intracellular signaling through reactive species of oxygen and nitrogen.}, number={3}, journal={CHEST}, author={Rochelle, LG and Fischer, BM and Krunkosky, TM and Wright, DT and Adler, KB}, year={1996}, month={Mar}, pages={S35–S39} }
 @article{li_adler_1996, title={Histamine stimulates secretion of mucin and activates phospholipase C in airway epithelium: evidence for independent pathways.}, volume={153}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Li, H. and Adler, K. B.}, year={1996}, pages={A539} }
 @article{dye_adler_richards_dreher_1996, title={Injury of rat tracheal epithelial cultures by exposure to residual oil fly ash (ROFA) involves generation of the hydroxyl radical.}, volume={153}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Dye, J. A. and Adler, K. B. and Richards, J. R. and Dreher, K. L.}, year={1996}, pages={A542} }
 @article{allen_mcgahan_ferrell_adler_fleisher_1996, title={Nitric oxide synthase inhibitors exert differential time-dependent effects on LPS-induced uveitis}, volume={62}, ISSN={["0014-4835"]}, DOI={10.1006/exer.1996.0003}, abstractNote={Nitric oxide (NO) is a highly reactive radical which plays an integral role in physiological and pathophysiological processes. NO is produced endogenously in small amounts by a constitutive NO synthase (cNOS) as a regulator of vascular tone and neurotransmission. NO can also be produced in large amounts by an inducible NOS (iNOS) in response to endotoxin and cytokines, and has been reported to be a mediator of lipopolysaccharide (LPS)-induced uveitis in rats. The purpose of the present study was to investigate the effects of NOS inhibitors with different NOS isoform specificities in the rabbit model of endotoxin-induced ocular inflammation. LPS and/or inhibitors of NOS. NG-nitro-L-arginine methyl ester (L-NAME) and aminoguanidine (AG), were injected intravitreally and the eyes observed by slit lamp for 24 hr. Coinjection of LPS with L-NAME inhibited anterior inflammation in rabbits. Iridal hyperemia (IH) and aqueous flare (AF) were completely abolished in eight out of nine rabbits in a dose-dependent manner. In addition, total cell counts were significantly suppressed (7393 +/- 697 vs. 325 +/- 188, P < 0.05) and aqueous protein levels were reduced to near control levels (25 +/- 0.75 vs. 1.72 +/- 0.36, P < 0.05). Similar suppression was seen with AG (cell counts = 351 +/- 246 and proteins = 3.1 +/- 1.2). Administration of L-NAME 0.5 hr after LPS injection suppressed inflammation to a lesser extent than coinjection. In contrast, administration of L-NAME 6 hr after LPS injection was not inhibitory, and in fact significantly increased cellular infiltration. However, AG given 6 hr after LPS had a remarkably different effect, since it significantly decreased both protein extravasation and cellular infiltration into the aqueous humor. In fact, our results suggest that cNOS may play a greater role in the earlier stages of this developing inflammatory response. These results extend others' observations that NO is a key mediator in uveitis, that induction of iNOS plays a critical role in experimental uveitis, and suggest that NO has a complex role in the ocular inflammatory process. Inhibitors of NOS can abort the LPS-induced inflammatory response if administered early enough, but could potentially exacerbate an established inflammatory episode.}, number={1}, journal={EXPERIMENTAL EYE RESEARCH}, author={Allen, JB and McGahan, MC and Ferrell, JB and Adler, KB and Fleisher, LN}, year={1996}, month={Jan}, pages={21–28} }
 @article{adler_wright_li_li_choe_rochell_fischer_1996, title={Nitric oxide: A key signaling molecule regulating airway mucin secretion.}, volume={9}, journal={Proceedings of the 9th World Congress on Bronchology & World Congress on Bronchoesophagology.}, author={Adler, K. B. and Wright, D. T. and Li, C. and Li, H. and Choe, N. and Rochell, L. G. and Fischer, B. M.}, year={1996}, pages={S060} }
 @article{wright_fischer_li_rochelle_akley_adler_1996, title={Oxidant stress stimulates mucin secretion and PLG in airway epithelium via a nitric oxide-dependent mechanism}, volume={271}, ISSN={["1040-0605"]}, DOI={10.1152/ajplung.1996.271.5.l854}, abstractNote={ Reactive oxygen species (ROS) have been implicated in the pathogenesis of a wide variety of respiratory diseases. We investigated mechanisms of ROS-induced mucin secretion by guinea pig tracheal epithelial (GPTE) cells in primary culture, and ROS-induced activation of the second messenger-producing enzyme phospholipase C (PLC), in GPTE cells and in a virally transformed cell line (BEAS-2B) derived from human bronchial epithelium. Mucin secretion was measured by a monoclonal antibody-based enzyme-linked immunosorbent assay, and PLC activation was assessed by anion exchange chromatography. ROS generated enzymatically by xanthine oxidase (XO, 500 microM) in the presence of purine (500 microM) enhanced release of mucin by GPTE cells and activated PLC in GPTE and BEAS cells. Hypersecretion of mucin and activation of PLC in response to purine + XO appeared to occur via an intracellular pathway(s) dependent on endogenously produced nitric oxide and possibly intracellularly generated oxidants. Both responses could be blocked or attenuated by preincubation of the cells with NG-monomethyl-L-arginine, an inhibitor of the enzyme nitric oxide synthase, or with dimethylthiourea, a compound that can react with a variety of intracellular oxidant species. Reactive nitrogen species generated chemically also stimulated secretion of mucin and activated PLC via a mechanism dependent (at least in part) on intracellular oxidant-mediated process(es). The results suggest that intracellularly generated radical species of nitrogen and oxygen may be important modulators of the response of airway epithelial cells to external oxidant stress. }, number={5}, journal={AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY}, author={Wright, DT and Fischer, BM and Li, CM and Rochelle, LG and Akley, NJ and Adler, KB}, year={1996}, month={Nov}, pages={L854–L861} }
 @article{adler_wright_fischer_1996, title={Reactive oxygen species provoke secretion of mucin and activate phospholipase C in airway epithelial cells in vitro via a mechanism dependent on nitric oxide and hydroxyl free radical.}, journal={Molecules to man, proceedings of Keystone Symposium}, author={Adler, K. B. and Wright, D. T. and Fischer, B. M.}, year={1996}, pages={20} }
 @article{adler_1996, title={Role of airway epithelium in airway inflammation.}, journal={Proceedings of the Symposium on Airway Mucus (Seoul, 1996).}, author={Adler, K. B.}, year={1996}, pages={32} }
 @article{adler_krunkosky_fischer_rochelle_martin_dreher_jiang_dye_1996, title={Role or reactive oxygen and nitrogen species in the response of airway epithelium to particulates.}, volume={6}, journal={Proceedings of the 6th International Meeting of the Toxicology of Natural and Man-Made Fibrous and Non-Fibrous Particles.}, author={Adler, K. B. and Krunkosky, T. M. and Fischer, B. M. and Rochelle, L. G. and Martin, L. D. and Dreher, K. L. and Jiang, N. and Dye, J.}, year={1996}, pages={139} }
 @article{krunkosky_fischer_akley_adler_1996, title={Tumor necrosis factor alpha (TNF alpha)-induced ICAM-1 surface expression in airway epithelial cells in vitro: Possible signal transduction mechanisms}, volume={796}, ISBN={["0-89766-979-7"]}, ISSN={["0077-8923"]}, DOI={10.1111/j.1749-6632.1996.tb32564.x}, abstractNote={Within the past several years research on the interaction of cytokines and adhesion molecules with airway epithelium in diseases has allowed us to develop a better understanding of the disease process. The cytokine, TNF alpha and the adhesion molecule ICAM-1 are important mediators in the pathogenesis of airway diseases such as asthma, chronic bronchitis, and adult respiratory distress syndrome. Effects of TNF alpha on ICAM-1 surface expression was investigated in both primary cultures of normal human bronchial epithelial (NHBE) cells and immortalized human bronchial epithelial cell line BEAS-2B. TNF alpha (0.015-150 ng/mL) significantly enhanced ICAM-1 surface expression (measured by flow cytometry) in a dose and time-dependent manner, with peak expression seen at 24 hours. This response was negated by heat inactivation of the TNF alpha prior to incubation. TNF alpha-induced ICAM-1 expression also was inhibited by pre- and coincubation of TNF alpha with 3 micrograms/mL soluble TNF-R1 or by the PKC inhibitor, Calphostin C (0.1 and 0.5 microM). The ROI scavengers, dimethylthiourea (4 mM), and dimethyl sulfoxide (0.001%), enhanced TNF alpha-induced ICAM-1 expression. Collectively, these results indicate that TNF alpha-induced ICAM-1 surface expression is a specific receptor-mediated response (TNF-R1), which is mediated by mechanisms dependent on PKC and intracellular reactive oxygen species.}, journal={CYTOKINES AND ADHESION MOLECULES IN LUNG INFLAMMATION}, author={Krunkosky, TM and Fischer, BM and Akley, NJ and Adler, KB}, year={1996}, pages={30–37} }
 @article{krunkosky_fischer_martin_adler_1996, title={Tumor necrosis factor alpha (TNF?) provokes expression of intercellular adhesion molecule-1 (ICAM-1) on human bronchial epithelial cells in primary culture.}, volume={153}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Krunkosky, T. M. and Fischer, B. M. and Martin, L. D. and Adler, K. B.}, year={1996}, pages={A27} }
 @article{fischer_rochelle_akley_adler_1996, title={Tumor necrosis factor-alpha (TNF?) induced mucin secretion: dependence on nitric oxide (NO) and cyclic GMP.}, volume={153}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Fischer, B. M. and Rochelle, L. G. and Akley, N. J. and Adler, K. B.}, year={1996}, pages={A538} }
 @article{lee_madden_reed_adler_devlin_1996, title={Use of the single cell gel electrophoresis assay (SCGE) for detection of oxidant-induced DNA single strand breaks (SSB) in human lung cells in vitro.}, volume={141}, DOI={10.1006/taap.1996.0276}, abstractNote={DNA single strand breaks (SSB) can be used as a biomarker ofoxidant exposure, and also as an indicator of the carcinogenicity/mutagenicity of a substance. The single cell gel electrophoresis (SCGE) assay is more sensitive and requires fewer cells compared to other techniques used for detecting SSB. We examined the utility of using the SCGE assay for human lung cells exposed to endogenous and exogenous oxidants. A human bronchial cell line (BEAS) was used as a model of airway epithelial cells in this study. BEAS cells exposed to 0–50 μM hydrogen peroxide (H2O2) for 60 min at 4°C exhibited a concentration-dependent increase in SSB as determined by an increased DNA migration area in a gel undergoing electrophoresis. H2O2-induced increases in DNA SSB were also demonstrated using cultured normal human tracheobronchial epithelial (NHBE) cells and human alveolar macrophages in a concentration response manner. BEAS cells were also exposed to air or ozone (O3) on a Transwell filter without medium present apically. Cells exposed to O3 at 0.1 or 0.4 ppm at 37°C for up 120 min had a time- and concentration-dependent increase in SSB compared to air-exposed cells. NHBE cells exposed to 0.4 ppm 03 (60 min) also had increased DNA SSB. Cells with H2O2-induced DNA SSB can be frozen and stored up to 4 weeks without altering the original DNA SSB. These findings indicate that SC GE can be used to detect SSB in cultured lung cells, and has applicability for detecting SSB in lung cells recovered from in vivo and in vitro exposures to oxidants.}, journal={Toxicology and Applied Pharmacology}, author={Lee, J. G. and Madden, M. C. and Reed, W. and Adler, K. B. and Devlin, R. B.}, year={1996}, pages={195–204} }
 @article{krunkosky_fischer_wright_akley_adler_1995, title={Effects of tumor necrosis factor alpha (TNF?) on expression of ICAM-1 and inflammation-associated genes in airway epithelial cells in vitro.}, volume={151}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Krunkosky, T. M. and Fischer, B. M. and Wright, D. T. and Akley, N. J. and Adler, K. B.}, year={1995}, pages={A366} }
 @article{fischer_wright_li_li_cohn_akley_adler_1995, title={Endogenously-generated nitric oxide (NO) may be a key signaling molecule in hypersecretion of mucin by guinea pig tracheal epithelial (GPTE) cells in vitro.}, volume={151}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Fischer, B. M. and Wright, D. T. and Li, H. and Li, C. M. and Cohn, L. A. and Akley, N. J. and Adler, K. B.}, year={1995}, pages={A337} }
 @article{li_choe_wright_adler_1995, title={HISTAMINE PROVOKES TURNOVER OF INOSITOL PHOSPHOLIPIDS IN GUINEA-PIG AND HUMAN AIRWAY EPITHELIAL-CELLS VIA AN H-1-RECEPTOR G-PROTEIN-DEPENDENT MECHANISM}, volume={12}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.12.4.7695921}, abstractNote={Guinea pig tracheal epithelial cells in primary air/liquid interface culture (GPTE) and virally transformed human bronchial epithelial cells (BEAS-2B) were exposed to histamine at concentrations of 1 to 100 microM. At concentrations greater than 1 microM, histamine elicited a concentration-dependent increase in accumulation of inositol phosphates in both cell types, as assessed by anion exchange chromatography. The effects of histamine were most pronounced at 15 to 30 min and were attenuated by the H1-receptor antagonist, pyrilamine. The H2-receptor antagonist, ranitidine, was without effect. Sodium fluoride (25 mM), a non-receptor-associated activator of GTP binding (G) proteins, increased accumulation of inositol phosphates within GPTE and BEAS cells. In cells permeabilized with digitonin, the nonhydrolyzable GTP analog, guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S; 10 microM) increased inositol phosphate accumulation. This GTP gamma S-induced increase was attenuated by exposure to 500 microM guanosine-5'-O-(2-thiodiphosphate) (GDP beta S). Additionally, histamine-induced increases in inositol phosphate accumulation were potentiated by GTP gamma S and attenuated by GDP beta S. These data indicate involvement of a G protein in the response to histamine. Preincubation with pertussis toxin (100 ng/ml for 4 h) did not significantly affect the response, suggesting that the associated G protein was not pertussis toxin-sensitive. The presence of the phosphatidylinositol-specific phospholipase C (PI-PLC)-associated G protein, G alpha q/11, and the presence of mRNA for the Gq family, were ascertained by immunoblotting and Northern hybridization, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)}, number={4}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={LI, HF and CHOE, NH and WRIGHT, DT and ADLER, KB}, year={1995}, month={Apr}, pages={416–424} }
 @article{adler_fischer_li_choe_wright_1995, title={HYPERSECRETION OF MUCIN IN RESPONSE TO INFLAMMATORY MEDIATORS BY GUINEA-PIG TRACHEAL EPITHELIAL-CELLS IN-VITRO IS BLOCKED BY INHIBITION OF NITRIC-OXIDE SYNTHASE}, volume={13}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb.13.5.7576687}, abstractNote={Primary cultures of guinea pig tracheal epithelial cells in air/liquid interface were exposed to one of four agents associated with airway inflammation: the peptide histamine (100 microM), the lipid mediator platelet-activating factor (1 microM), the cytokine tumor necrosis factor-alpha (15 ng/ml; specific activity 2.86 x 10(7) U/mg), or enzymatically generated reactive oxygen species (purine [500 microM]+xanthine oxidase [20 mU/ml]). Effects of each of these substances on release of mucin by guinea pig tracheal epithelial (GPTE) cells were measured using a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA). Each secretagogue significantly enhanced release of mucin, but the stimulatory effect of each was inhibited by pre-(+)co-incubation of the cells with the competitive inhibitor of nitric oxide synthase, NG-monomethyl-L-arginine (L-NMA), but not by NG-monomethyl-D-arginine (D-NMA), the inactive stereoisomer that does not inhibit nitric oxide synthase. Neither L-NMA nor D-NMA affected mucin secretion by themselves. The results suggest that each of these inflammation-associated mediators provokes airway epithelial mucin secretion via a mechanism involving intracellular production of nitric oxide (NO) as a critical signaling molecule.}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={ADLER, KB and FISCHER, BM and LI, HF and CHOE, NH and WRIGHT, DT}, year={1995}, month={Nov}, pages={526–530} }
 @article{lee_madden_hatch_bottei_reed_adler_devlin_1995, title={Ozone exposure in vivo induces increases in DNA single strand breaks in human and guinea pig lung cells.}, volume={151}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Lee, J. G. and Madden, M. C. and Hatch, G. E. and Bottei, G. and Reed, W. and Adler, K. B. and Devlin, R. B.}, year={1995}, pages={A498} }
 @article{wright_li_fischer_akley_adler_1995, title={Reactive nitrogen species (RNS) stimulate secretion of mucin by primary cultures of guinea pig tracheal epithelial (GPTE) cells.}, volume={151}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Wright, D. T. and Li, C. M. and Fischer, B. M. and Akley, N. J. and Adler, K. B.}, year={1995}, pages={A337} }
 @article{fischer_krunkosky_wright_dolan-o'keefe_adler_1995, title={Tumor necrosis factor alpha (TNF?) stimulates mucin secretion and gene expression in airway epithelium invitro.}, volume={107}, DOI={10.1378/chest.107.3_supplement.133s}, abstractNote={TNF-α=Tumor necrosis factor α mRNA=messenger RNA; NO=nitric oxide; NOS=nitric oxide synthase; iNOS=calcium independent nitric oxide synthase; GPTE=guinea pig tracheal epithelial; OD=optical density; ELISA=enzyme-linked immunosorbent assay; mAb=monoclonal antibody; BSA=bovine serum albumin; PBS=phosphate buffered saline solution; FBS=fetal bovine serum; rhTNF-α=recombinant human tumor necrosis factor α PC-PLC=phosphatidylcholine-specific phospholipase; CPKC=protein kinase C; DAG=diacylglycerol; PLA2=phospholipase A2; cPLA2=cytosolic phospholipase A2; MnSOD=manganese dependent superoxide dismutase; L-NMA=L-NG-monomethyl arginine; SNAP=S-nitroso-N-acetylpenacil-lamine; IL-6=interleukin-6; NF-κ β=nuclear factor kappa beta TNF-α=Tumor necrosis factor α mRNA=messenger RNA; NO=nitric oxide; NOS=nitric oxide synthase; iNOS=calcium independent nitric oxide synthase; GPTE=guinea pig tracheal epithelial; OD=optical density; ELISA=enzyme-linked immunosorbent assay; mAb=monoclonal antibody; BSA=bovine serum albumin; PBS=phosphate buffered saline solution; FBS=fetal bovine serum; rhTNF-α=recombinant human tumor necrosis factor α PC-PLC=phosphatidylcholine-specific phospholipase; CPKC=protein kinase C; DAG=diacylglycerol; PLA2=phospholipase A2; cPLA2=cytosolic phospholipase A2; MnSOD=manganese dependent superoxide dismutase; L-NMA=L-NG-monomethyl arginine; SNAP=S-nitroso-N-acetylpenacil-lamine; IL-6=interleukin-6; NF-κ β=nuclear factor kappa beta}, journal={Chest}, author={Fischer, B. M. and Krunkosky, T. M. and Wright, D. T. and Dolan-O'Keefe, M. and Adler, K. B.}, year={1995}, pages={133S–135} }
 @inbook{cohn_adler_1994, title={Alternatives to in vivo toxicological testing of rodent airway epithelia.}, booktitle={Animal test alternatives: refinement-reduction-replacement.}, author={Cohn, L. A. and Adler, K. B.}, year={1994}, pages={233–239} }
 @article{cohn_adler_1994, title={Antioxidant scavenging by guinea pig and human airway epithelial cells in vitro: mechanisms of H2O2 clearance.}, volume={265}, number={10}, journal={American Journal of Physiology. Lung Cellular and Molecular Physiology}, author={Cohn, L. A. and Adler, K. B.}, year={1994}, pages={397–404} }
 @article{dye_adler_1994, title={Effects of cigarette smoke on epithelial cells of the respiratory tract.}, volume={49}, DOI={10.1136/thx.49.8.825}, abstractNote={Chronic inhalation of cigarette smoke is associated with mucus hypersecretion, mucus pooling, pulmonary connective tissue damage, and chronic airflow obstruction. Cigarette smoke has therefore been causally linked to the development of chronic obstructive pulmonary disease (either chronic bronchitis or emphysema),' increased airway responsiveness,2 exacerbations of asthma,3 impaired pulmonary immune function,4 and increased pulmonary infections.5 Cigarette smoke has also been established as an important risk factor for lung,6 laryngeal,7 and nasal neoplasia.5 Pathogenetic mechanisms related to smoke-induced respiratory perturbations, however, are not fully understood. One cell type in the lung that may play a major part in the pathogenesis of cigarette smoke-induced lesions is the airway epithelial cell. These cells line the lumen of the airways, and thus are in a unique position to interact directly with inhaled cigarette smoke. Most research involving cigarette smoke and airway epithelial cells has focused on the "target" cell responses of these cells in relation to their relatively simple roles in barrier and mucociliary clearance functions. Depending in part on the chronicity of exposure, certain functions may be altered for example, ciliary beating, mucus secretion. It has recently become apparent, however, that airway epithelial cells may also act as "effector" cells, playing pivotal roles in regulation of airway reflexes, immunological and inflammatory responses, and maintenance of bronchodilation. As part of their overall response to chronic insult these cells are capable of producing and/or releasing a number of inflammatory mediators, or undergoing alterations in expression of cell adhesion molecules processes that may initiate or perpetuate airway inflammation.9 To date the influence of cigarette smoke on effector functions of epithelial cells has yet to be investigated in detail. Much of the information presented herein is based on acute in vitro cigarette smoke exposures of epithelial cell cultures or airway explants, and on relatively acute human or laboratory animal exposures. Thus, non-neoplastic and non-emphysematous end points of respiratory disease have been emphasised. Nevertheless, early events in the response to cigarette smoke or its components may be critical, and certainly an understanding of these events may help to elucidate the pathogenetic mechanisms of many chronic respiratory diseases. Components of cigarette smoke Chemical analytical studies have identified over 3800 compounds in tobacco smoke.'" Mainstream cigarette smoke is composed of a complex mixture of gases and condensed tar particles. In experimental studies cigarette smoke is often separated into two phases by a glass fibre filter that retains nearly all particulate matter greater than 01 ptm in diameter. The retained particulate matter is commonly referred to as the "tar" phase, while the material passing through the filter is referred to as the "gas" phase. Known toxins and carcinogens have been identified in both the gaseous and particulate phases." Sidestream smoke (smoke emitted from the burning tip of the cigarette) is the major constituent of environmental tobacco smoke. The chemical composition and gas-toparticle associations of environmental tobacco smoke may be different from that of mainstream smoke, owing to prolonged time and cooling in the air.'2 Sidestream cigarette smoke emissions contain carbon monoxide, ammonia, formaldehyde, benzene, nicotine, acrolein, various gases and particles, and an assortment of potentially genotoxic and/or carcinogenic organic compounds.'3 Increased pulmonary particulate burden due to cigarette smoke may also play a part in respiratory disease.'4 Recent epidemiological findings have indicated adverse effects of particulate air pollutants at concentrations below currently permissible levels.'5 Respirable suspended particles in indoor air in homes may increase from approximately 30 jtg/ m3 to greater than 60 jg/M3 due to accumulation of environmental tobacco smoke.'6 The reported effects of "cigarette smoke" may include that of mainstream smoke, variably aged environmental tobacco smoke/sidestream and exhaled smoke, gaseous phase components only, particulate phase components only, or individual chemical compounds such as acrolein,'7 acetaldehyde,'5 or formaldehyde.'9 Some studies have used aqueous extracts of cigarette smoke obtained by bubbling the smoke through a buffer, with or without filtering to remove suspended particulates,20 while other researchers have focused on free radical production arising from chemical reactions within the cigarette smoke.2' Overall, owing to the variability in experimental methodologies (including the type of "cigarette smoke") used, interstudy comparisons may be difficult to interpret. It is well to keep this in mind when reading this review. Department of Anatomy, Physiological Sciences, and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27606, USA J A Dye K B Adler}, journal={404nOtfound}, author={Dye, J. A. and Adler, K. B.}, year={1994}, pages={825–834} }
 @article{adler_wright_fischer_cohn_li_li_choe_akley_1994, title={Effects of oxidant stress on airway epithelial cells in vitro.}, volume={8}, journal={FASEB Journal}, author={Adler, K. B. and Wright, D. T. and Fischer, B. M. and Cohn, L. A. and Li, C. and Li, H. and Choe, N. H. and Akley, N. J.}, year={1994}, pages={A896} }
 @article{adler_fischer_wright_dolan-o'keefe_1994, title={Effects of tumor necrosis factor alpha (TNF?) on airway epithelial cells in vitro.}, volume={8}, journal={Proceedings of the 8th International Colloquium on Lung Fibrosis}, author={Adler, K. B. and Fischer, B. M. and Wright, D. T. and Dolan-O'Keefe, M.}, year={1994}, pages={3} }
 @article{adler_fischer_wright_cohn_becker_1994, title={INTERACTIONS BETWEEN RESPIRATORY EPITHELIAL-CELLS AND CYTOKINES - RELATIONSHIPS TO LUNG INFLAMMATION}, volume={725}, ISBN={["0-89766-856-1"]}, ISSN={["0077-8923"]}, DOI={10.1111/j.1749-6632.1994.tb00275.x}, abstractNote={Epithelial cells lining respiratory airways can participate in inflammation in a number of ways. They can act as target cells, responding to exposure to a variety of inflammatory mediators and cytokines by altering one or several of their functions, such as mucin secretion, ion transport, or ciliary beating. Aberrations in any of these functions can affect local inflammatory responses and compromise pulmonary defense. For example, oxidant stress can increase secretion of mucin and depress ciliary beating efficiency, thereby affecting the ability of the mucociliary system to clear potentially pathogenic microbial agents. Recent studies have indicated that airway epithelial cells also can act as "effector" cells, synthesizing and releasing cytokines, lipid mediators, and reactive oxygen species in response to a number of pathologically relevant stimuli, thereby contributing to inflammation. Many of these epithelial-derived substances can act locally, affecting both neighboring cells and tissues, or, via autocrine or paracrine mechanisms, affect structure and function of the epithelial cells themselves. Studies in our laboratories utilized cell cultures of both human and guinea pig tracheobronchial and nasal epithelial cells, and isolated human nasal epithelial cells, to investigate activity of respiratory epithelial cells in vitro as sources of cytokines and inflammatory mediators. Primary cultures of guinea pig and human tracheobronchial and nasal epithelial cells synthesize and secrete low levels of IL-6 and IL-8 constitutively. Production and release of these cytokines increases substantially after exposure to specific inflammatory stimuli, such as TNF or IL-1, and after viral infection.}, journal={CELLS AND CYTOKINES IN LUNG INFLAMMATION}, author={ADLER, KB and FISCHER, BM and WRIGHT, DT and COHN, LA and BECKER, S}, year={1994}, pages={128–145} }
 @article{wright_cohn_li_fischer_li_adler_1994, title={Interactions of oxygen radicals with airway epithelium.}, volume={102}, DOI={10.1289/ehp.94102s1085}, abstractNote={Reactive oxygen species (ROS) have been implicated in the pathogenesis of numerous disease processes. Epithelial cells lining the respiratory airways are uniquely vulnerable regarding potential for oxidative damage due to their potential for exposure to both endogenous (e.g., mitochondrial respiration, phagocytic respiratory burst, cellular oxidases) and exogenous (e.g., air pollutants, xenobiotics, catalase negative organisms) oxidants. Airway epithelial cells use several nonenzymatic and enzymatic antioxidant mechanisms to protect against oxidative insult. Nonenzymatic defenses include certain vitamins and low molecular weight compounds such as thiols. The enzymes superoxide dismutase, catalase, and glutatione peroxidase are major sources of antioxidant protection. Other materials associated with airway epithelium such as mucus, epithelial lining fluid, and even the basement membrane/extracellular matrix may have protective actions as well. When the normal balance between oxidants and antioxidants is upset, oxidant stress ensues and subsequent epithelial cell alterations or damage may be a critical component in the pathogenesis of several respiratory diseases. Oxidant stress may profoundly alter lung physiology including pulmonary function (e.g., forced expiratory volumes, flow rates, and maximal inspiratory capacity), mucociliary activity, and airway reactivity. ROS may induce airway inflammation; the inflammatory process may serve as an additional source of ROS in airways and provoke the pathophysiologic responses described. On a more fundamental level, cellular mechanisms in the pathogenesis of ROS may involve activation of intracellular signaling enzymes including phospholipases and protein kinases stimulating the release of inflammatory lipids and cytokines. Respiratory epithelium may be intimately involved in defense against, and pathophysiologic changes invoked by, ROS.}, number={11}, journal={Environmental Health Perspectives. Supplements}, author={Wright, D. T. and Cohn, L. A. and Li, H. and Fischer, B. M. and Li, C. M. and Adler, K. B.}, year={1994}, pages={85–90} }
 @article{wright_friedman_pryor_squadrito_salgo_adler_1994, title={Lipid ozonation products activate phospholipases in guinea pig and human airway epithelial cells in vitro.}, volume={149}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Wright, D. T. and Friedman, M. and Pryor, W. A. and Squadrito, G. L. and Salgo, M. G. and Adler, K. B.}, year={1994}, pages={A320} }
 @article{wright_adler_akley_dailey_friedman_1994, title={OZONE STIMULATES RELEASE OF PLATELET-ACTIVATING-FACTOR AND ACTIVATES PHOSPHOLIPASES IN GUINEA-PIG TRACHEAL EPITHELIAL-CELLS IN PRIMARY CULTURE}, volume={127}, ISSN={["1096-0333"]}, DOI={10.1006/taap.1994.1135}, abstractNote={Inhalation of ozone (O3) has been associated with development of inflammation in the respiratory airways and a variety of alterations in pulmonary function. Epithelial cells lining the airways are the first cells with which inhaled O3 comes into contact and thus represent a potential major target of acute O3 toxicity. In addition, upon appropriate stimulation or injury, these cells are capable of releasing a spectrum of secondary mediators that could relate to the pathogenesis of O3-associated lesions. We exposed organotypically cultured guinea pig primary tracheal epithelial (GPTE) cells in an air/liquid interface to photochemically generated O3 in vitro and monitored effects of O3 exposure on activation of phospholipases A2 (PLA2), C (PLC), and D (PLD), as well as release of the humoral mediator, platelet activating factor (PAF). PLA2 acts on ether-linked phosphatidylcholine, which upon further metabolism forms PAF;PLC acts on inositol phospholipids to produce inositol phosphates and diacylglycerol; and PLD generates phosphatidic acid. GPTE cell cultures exposed to O3 (0.05-1.0 ppm) for 1 hr displayed an elevated total release of PAF (apical+basolateral). Maximal stimulation in both apical and total release of PAF occurred at 1.0 ppm O3 (405 +/- 47 and 282 +/- 23% of air control values, respectively, n = 7). The 1.0 ppm O3-induced increased PAF release was significantly inhibitable by the PLA2 inhibitor mepacrine (1 mM), suggesting a connection between PAF release and PLA2 activation. O3 exposure activated PLC in GPTE cells in a concentration- (0.1-1.0 ppm) and time-dependent (10-60 min) manner to produce a significant accumulation of inositol-1,4,5-triphosphate, with maximal accumulation at 1.0 ppm O3 for 1 hr (417 +/- 121% of air control, n = 6). PAF receptor antagonists Ro 24-4736 (1 microM) and Ro 41-5036 (1 microM) did not affect O3-stimulated inositol phosphate accumulation. PLD also was activated in GPTE cells exposed to 1.0 ppm O3 for 1 hr (169 +/- 80% of air control, n = 5). These results suggest that GPTE cells respond to O3 exposure in vitro by increasing production and/or release of PAF via a mechanism that may involve activation of PLA2, PLC, and PLD. Epithelial-derived mediators, such as PAF, may play a role in the pathogenesis of lesions associated with inhalation of O3.}, number={1}, journal={TOXICOLOGY AND APPLIED PHARMACOLOGY}, author={WRIGHT, DT and ADLER, KB and AKLEY, NJ and DAILEY, LA and FRIEDMAN, M}, year={1994}, month={Jul}, pages={27–36} }
 @article{li_cheng_adler_1994, title={PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST GUINEA-PIG TRACHEAL MUCINS}, volume={13}, ISSN={["0272-457X"]}, DOI={10.1089/hyb.1994.13.281}, abstractNote={Thirty-five hybridomas that secrete mouse monoclonal antibodies (MAb) against guinea pig (G.P.) tracheal mucins were established. The MAbs were characterized immunologically, biochemically, and immunohistochemically at both light and electron microscopic levels. Isotyping of the MAbs revealed 14 to be IgM, 13 IgG1, 3 IgG2, and 5 IgG3. The MAbs demonstrated various patterns of binding in immunoblots against mucins derived from G.P. tracheal explants. This suggested the presence of "subpopulations" of G.P. tracheal mucins with specific MAbs binding to different epitopes on the mucin molecules. Periodate oxidation indicated that 33 of the 35 MAbs recognized carbohydrate epitopes on the mucin molecules. Ten of the MAbs also reacted with both bovine and ferret tracheal mucins, while 7 and 6 MAbs bound only to bovine and ferret tracheal mucins, respectively. The generated MAbs should be useful for immunomeasurement of mucin secretion in vivo (e.g., in bronchoalveolar or airway lavage fluid) and in vitro (e.g., cell and organ cultures) from cells of guinea pig and (with certain MAbs) bovine and ferret origin.}, number={4}, journal={HYBRIDOMA}, author={LI, CM and CHENG, PW and ADLER, KB}, year={1994}, month={Aug}, pages={281–287} }
 @article{choe_li_adler_1994, title={Platelet activating factor and histamine activate phospholipase C in  airway epithelial cells in vitro via a pertussis toxin sensitive G protein.}, volume={149}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Choe, N.-H. and Li, H. and Adler, K. B.}, year={1994}, pages={A985} }
 @article{wright_akley_li_fischer_adler_1994, title={Reactive oxygen species (ROS) activate phospholipase C in guinea pig and human airway epithelial cells in vitro.}, volume={149}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Wright, D. T. and Akley, N. J. and Li, H. and Fischer, B. M. and Adler, K. B.}, year={1994}, pages={A315} }
 @article{adler_fischer_akley_dolan-o'keefe_1994, title={Tumor necrosis factor alpha (TNF?) stimulates mucin secretion and gene expression in airway epithelium in vitro.}, volume={37}, journal={Proceedings of the 37th Aspen Lung Conference.}, author={Adler, K. B. and Fischer, B. M. and Akley, N. J. and Dolan-O'Keefe, M.}, year={1994}, pages={17} }
 @article{lee_madden_reed_adler_devlin_1994, title={Use of the single cell gel electrophoresis assay (SCGE) for detection of oxidant-induced DNA single strand breaks (SSB) in human lung cells.}, volume={149}, journal={American Journal of Respiratory and Critical Care Medicine}, author={Lee, J. G. and Madden, M. C. and Reed, W. and Adler, K. B. and Devlin, R. B.}, year={1994}, pages={A158} }
 @article{cohn_adler_1993, title={Antioxidant scavenging by guinea pig and human airway epithelial cells in vitro: mechanisms of H2O2 clearance.}, volume={147}, journal={American Review of Respiratory Disease}, author={Cohn, L. A. and Adler, K. B.}, year={1993}, pages={A440} }
 @article{kaartinen_nettesheim_adler_randell_1993, title={RAT TRACHEAL EPITHELIAL-CELL DIFFERENTIATION IN-VITRO}, volume={29A}, ISSN={["1543-706X"]}, DOI={10.1007/bf02639383}, abstractNote={In vitro culture conditions enabling rat tracheal epithelial (RTE) cells to differentiate to mucociliary, mucous, or squamous phenotypes are described. Medium composition for rapid cell growth to confluence in membrane insert cultures was determined, and the effects of major modifiers of differentiation were tested. Retinoic acid (RA), collagen gel substratum, and an air-liquid interface at the level of the cell layer were required for expression of a mucociliary phenotype which most closely approximated the morphology of the tracheal epithelium in vivo. Large quantities of high molecular weight, hyaluronidase-resistant glycoconjugates, most likely mucin glycoproteins, were produced in the presence of RA when the cells were grown with or without a collagen gel and in submerged as well as in interface cultures. However, extensive ciliagenesis was dependent on the simultaneous presence of RA, collagen gel, and an air-liquid interface. When RA was omitted from the media, the cells became stratified squamous and developed a cornified apical layer in air-liquid interface cultures. This phenotype was accompanied by loss of transglutaminase (TGase) type II and keratin 18 and expression of the squamous markers TGase type I and keratin 13. The ability to modulate RTE cell phenotypes in culture will facilitate future studies investigating molecular regulation of tracheal cell proliferation, differentiation, and function.}, number={6}, journal={IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL}, author={KAARTINEN, L and NETTESHEIM, P and ADLER, KB and RANDELL, SH}, year={1993}, month={Jun}, pages={481–492} }
 @inbook{cohn_akley_adler_1993, title={Study of xenobiotics and airway epithelium in vitro.}, booktitle={Fiber toxicology.}, author={Cohn, L. A. and Akley, N. J. and Adler, K. B.}, year={1993}, pages={171–194} }
 @article{adler_cohn_1993, title={Use of air/liquid interface cultures to assess polarized responses of airway epithelium to toxicants.}, volume={29A}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Adler, K. B. and Cohn, L. A.}, year={1993}, pages={46A} }
 @inproceedings{adler_1993, title={Use of air/liquid interface guinea pig tracheal epithelial cell culture system for assessment of interactions between inhaled substances and airways.}, volume={1993}, number={Winter}, booktitle={Proceedings of the Toxicol Forum 1993 Annual Winter Meeting}, author={Adler, K. B.}, year={1993}, pages={14–19} }
 @article{adler_cohn_1993, title={Use of air/liquid interface guinea pig tracheal epithelial cell culture systems for assessment of interactions between inhaled substances and airways.}, volume={1993}, journal={Proceedings of the Toxicology Forum 1993.}, author={Adler, K. B. and Cohn, L. A.}, year={1993} }
 @article{rippetoe_absher_makrides_adler_evans_1992, title={Airway epithelial cells release factor(s) that stimulate proliferation of airway smooth muscle cells in vitro.}, volume={3}, journal={Molecular Biology of the Cell}, author={Rippetoe, P. E. and Absher, M. and Makrides, W. and Adler, K. B. and Evans, J. N.}, year={1992}, pages={175a} }
 @article{adler_cohn_1992, title={Guinea pig tracheal epithelial cells in primary air/liquid interface culture scavenge H2O2 via a mechanism only partially dependent on catalase and the glutathione system.}, volume={6}, journal={FASEB Journal}, author={Adler, K. B. and Cohn, L. A.}, year={1992}, pages={A939} }
 @article{adler_kinnula_akley_lee_cohn_crapo_1992, title={INFLAMMATORY MEDIATORS AND THE GENERATION AND RELEASE OF REACTIVE OXYGEN SPECIES BY AIRWAY EPITHELIUM INVITRO}, volume={101}, ISSN={["0012-3692"]}, DOI={10.1378/chest.101.3_supplement.53s}, abstractNote={Epithelial cells lining the respiratory airways represent the first cells with which inhaled inorganic particulates and microbes come into contact upon inhalation, yet surprisingly, little is known about interactions between these substances and epithelial cells. Most studies of such interactions relate to airway epithelial response as a classical “target” type of cell, responding to exogenous stimuli or related local inflammatory reactions by altering one or more facets of epithelial function, such as mucin secretion, ion transport, and ciliary beating. 1 Phipps RJ Denas SM Sielczak MW Wanner A. Effects of 0.5 ppm ozone on glycoprotein secretion, ion, and water fluxes in sheep trachea. J Appl Physiol. 1986; 60: 918-927 PubMed Google Scholar}, number={3}, journal={CHEST}, author={ADLER, KB and KINNULA, VL and AKLEY, N and LEE, JW and COHN, LA and CRAPO, JD}, year={1992}, month={Mar}, pages={S53–S54} }
 @misc{cohn_adler_1992, title={INTERACTIONS BETWEEN AIRWAY EPITHELIUM AND MEDIATORS OF INFLAMMATION}, volume={18}, ISSN={["0190-2148"]}, DOI={10.3109/01902149209031687}, abstractNote={Epithelial cells lining the respiratory airways classically are considered to be "target" cells, responding to exposure to a variety of inflammatory mediators by altering one or several of their functions, such as mucin secretion, ion transport, or ciliary beating. Specific responses of epithelial cells in vivo or in vitro to many of these inflammatory mediators are discussed. Recent studies have indicated that airway epithelial cells also can act as "effector" cells, responding to a variety of exogenous and/or endogenous stimuli by generating and releasing additional mediators of inflammation, such as eicosanoids, reactive oxygen species, and cytokines. Many of these epithelial-derived substances can diffuse away and affect neighboring cells and tissues, or can act, via autocrine or paracrine mechanisms, to affect structure and function of epithelial cells themselves. Studies dealing with airway epithelium as a source of inflammatory mediators and related compounds also are discussed.}, number={3}, journal={EXPERIMENTAL LUNG RESEARCH}, author={COHN, LA and ADLER, KB}, year={1992}, pages={299–322} }
 @article{friedman_wright_dailey_akley_devlin_adler_1992, title={Ozone increases platelet activating factor and activates phospholipases (PLA2 and PLC) in primary guinea pig tracheal epithelial cells (GPTE).}, volume={145}, journal={American Review of Respiratory Disease}, author={Friedman, M. and Wright, D. T. and Dailey, L. A. and Akley, N. J. and Devlin, R. B. and Adler, K. B.}, year={1992}, pages={A99} }
 @article{adler_akley_glasgow_1992, title={PLATELET-ACTIVATING-FACTOR PROVOKES RELEASE OF MUCIN-LIKE GLYCOPROTEINS FROM GUINEA-PIG RESPIRATORY EPITHELIAL-CELLS VIA A LIPOXYGENASE-DEPENDENT MECHANISM}, volume={6}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb/6.5.550}, abstractNote={Primary cultures of guinea pig tracheal epithelial cells maintained in an air/liquid interface system that maintains differentiated characteristics were grown to near confluence and exposed for 1 h to platelet-activating factor (PAF) on both apical and basal sides. PAF provoked release of high-molecular-weight mucin-like glycoproteins (MLG) from the cells, with maximal stimulation occurring at 10(-8) and 10(-9) M. The inactive form of PAF, lyso-PAF, was without effect. Indomethacin, the cyclooxygenase inhibitor, did not affect secretion stimulated by PAF, but nordihydroguiaretic acid (NDGA), a mixed cyclooxygenase and lipoxygenase inhibitor, attenuated secretion stimulated by PAF in a concentration-dependent manner. High performance liquid chromatography assay of the culture medium after addition of PAF revealed increased production of 15-, 12-, and 5-hydroxyeicosatetraenoic acids (15-, 12-, and 5-HETEs). The stimulatory effect of PAF on both mucin secretion and formation of HETEs was inhibited by the PAF receptor antagonists, CV-3988 and Ro 19 3704, with Ro 19 3704 acting at a concentration 10-fold lower than CV-3988 in inhibiting both effects. When added exogenously to the cell cultures, the combination of 5-, 12-, and 15-HETEs stimulated MLG release in a concentration-dependent manner. The results suggest that PAF stimulates release of MLG by guinea pig airway epithelium in vitro by a mechanism involving binding of PAF to receptors on epithelial cell surfaces, stimulation of lipoxygenase metabolism of arachidonic acid to HETEs within the epithelium, and stimulation of secretion by these epithelial-derived HETEs via an autocrine or paracrine mechanism.}, number={5}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={ADLER, KB and AKLEY, NJ and GLASGOW, WC}, year={1992}, month={May}, pages={550–556} }
 @article{kinnula_adler_ackley_crapo_1992, title={RELEASE OF REACTIVE OXYGEN SPECIES BY GUINEA-PIG TRACHEAL EPITHELIAL-CELLS INVITRO}, volume={262}, ISSN={["0002-9513"]}, DOI={10.1152/ajplung.1992.262.6.l708}, abstractNote={ Regulatory and stimulatory mechanisms of H2O2 release from guinea pig tracheal epithelial cells were investigated. Cells in primary culture maintained in a previously described air-liquid interface system released H2O2 to the extracellular space only from the apical side of the cells. The rate of release was 0.044 +/- 0.003 nmol.min-1.mg protein-1. H2O2 release could be stimulated significantly during a 30-min incubation period with phorbol myristate acetate (PMA) and platelet-activating factor (PAF). A stimulatory effect of PAF was achieved at concentrations greater than 100 nM and with PMA at concentrations greater than 10 ng (16 nM). When protein kinase C was inactivated with staurosporine, the responses to both PAF and PMA were abolished, whereas the cyclooxygenase inhibitor, indomethacin, did not affect H2O2 generation. When guinea pig tracheal epithelial cells were exposed to sublethal concentrations of extracellular H2O2 (30 microM), H2O2 was detoxified from both apical and basal sides, H2O2 removal being significantly more rapid from the apical side of the cells. These results suggest that tracheal epithelial cells can be stimulated to generate reactive oxygen species into the airway lumen and that this occurs in response to inflammatory mediators that act through protein kinase C. Luminal H2O2 release may have developed as a defense mechanism against microbes, and, similarly, luminal detoxification of H2O2 could represent an important mechanism of modulation of airway inflammation in response to oxidant stress. }, number={6}, journal={AMERICAN JOURNAL OF PHYSIOLOGY}, author={KINNULA, VL and ADLER, KB and ACKLEY, NJ and CRAPO, JD}, year={1992}, month={Jun}, pages={L708–L712} }
 @article{adler_kinnula_akley_crapo_1991, title={Airway epithelium generates and releases reactive oxygen species in response to activation of protein kinase C.}, volume={4}, journal={Proceedings fo the 4th International Conference on Environmental Lung Disease (Montreal, 1991)}, author={Adler, K. B. and Kinnula, V. and Akley, N. J. and Crapo, J. D.}, year={1991}, pages={2} }
 @article{mossman_janssen_marsh_sesko_shatos_doherty_adler_hemenway_mickey_vacek_et al._1991, title={DEVELOPMENT AND CHARACTERIZATION OF A RAPID-ONSET RODENT INHALATION MODEL OF ASBESTOSIS FOR DISEASE PREVENTION}, volume={19}, ISSN={["0192-6233"]}, DOI={10.1177/0192623391019004-110}, abstractNote={ A short-term inhalation model of asbestosis was developed in rodents to examine possible preventive approaches to lung disease. Fischer 344 (F344) rats were exposed for 10 and 20 days to National Institute of Environmental Health Sciences (NIEHS) crocidolite asbestos while sham controls were exposed to air only. To determine quantitative biochemical indicators of asbestos-induced lung disease, bronchoalveolar lavage (BAL) fluids were analyzed for lactic dehydrogenase (LDH), alkaline phosphatase, angiotensin-converting enzyme (ACE), and protein. Total and differential cell counts were performed on cell pellets from BAL. Lungs from additional rats were processed for histopathology, measurement of hydroxyproline, and autoradiography after injection of rats with 3H-thymidine. Exposure to asbestos for 10 and 20 days caused increases in LDH, alkaline phosphatase, and protein in BAL. In contrast, ACE was undetectable in BAL fluids from sham or asbestos-exposed rats. At both time periods, the percentages of polymorphonuclear leukocytes (PMNs) and lymphocytes in BAL were increased in asbestos-exposed rats. Total cell numbers in BAL were increased significantly at 20 days in animals inhaling asbestos. Exposure to asbestos for 10 and 20 days caused elevated amounts of hydroxyproline in lung and the development of fibrotic lesions. Asbestos-exposed rats exhibited increased numbers of interstitial cells and airspace epithelial cells incorporating 3H-thymidine, whereas labeled bronchiolar epithelial cells were not elevated significantly. The quantitative changes in asbestos-associated enzyme levels, cell types and protein in BAL, as well as increases in hydroxyproline and morphologic evidence of fibrosis, are useful indices of asbestos-related lung injury which enable preventive and therapeutic approaches to disease. }, number={4}, journal={TOXICOLOGIC PATHOLOGY}, author={MOSSMAN, BT and JANSSEN, YM and MARSH, JP and SESKO, A and SHATOS, MA and DOHERTY, J and ADLER, KB and HEMENWAY, D and MICKEY, R and VACEK, P and et al.}, year={1991}, pages={412–418} }
 @inbook{adler_henke_1991, title={Effects of inflammatory mediators on epithelial function and integrity.}, booktitle={Lung biology in health and disease.}, author={Adler, K. B. and Henke, D. C.}, year={1991}, pages={377–402} }
 @article{adler_kinnula_akley_lee_crapo_1991, title={Effects of inflammatory mediators on generation and release of reactive oxygen species by airway epithelium in vitro.}, volume={34}, journal={Proceedings of the 34th Aspen Lung Conference.}, author={Adler, K. B. and Kinnula, V. and Akley, N. J. and Lee, J. and Crapo, J. D.}, year={1991}, pages={33} }
 @article{cohn_adler_1991, title={INVITRO STUDIES OF MECHANISMS OF LUNG INJURY IN THE RODENT}, volume={19}, ISSN={["0192-6233"]}, DOI={10.1177/0192623391019004-111}, abstractNote={ In order to better define the responses of lung cells to potentially pathogenic insults, primary cell cultures of dissociated respiratory epithelial cells have been established. These epithelial cells have been obtained from various areas of the respiratory tract ranging from the trachea to the alveolus and the cultures have been demonstrated to mimic the differentiated state of these cell types as observed in situ. Several procedures which enhance the differentiated state have been evaluated, which include maintenance on more physiologically-relevant substrata, such as collagen gels, use of defined serum-free medium and use of air/liquid interface systems. These approaches have allowed intracellular responses of respiratory epithelium to toxic insult to be better defined. }, number={4}, journal={TOXICOLOGIC PATHOLOGY}, author={COHN, LA and ADLER, KB}, year={1991}, pages={419–427} }
 @article{adler_cohn_1991, title={In vitro studies on mechanisms of lung injury in the rodent.}, volume={10}, journal={Proceedings of the 10th International Symposium of Society of Toxicological Pathologists}, author={Adler, K. B. and Cohn, L. A.}, year={1991} }
 @inbook{lee_akley_adler_1991, title={Methods to study interactions between inhaled substances and epithelium of the respiratory tract in vitro.}, booktitle={Microbeam analysis- 1990.}, author={Lee, J. and Akley, N. J. and Adler, K. B.}, year={1991}, pages={457–458} }
 @article{kaartinen_nettesheim_adler_randell_1991, title={Modulation of rat tracheal epithelial cell differentiation in culture.}, volume={143}, journal={American Review of Respiratory Disease}, author={Kaartinen, L. and Nettesheim, P. and Adler, K. B. and Randell, S. H.}, year={1991}, pages={A146} }
 @article{glasgow_akley_adler_1991, title={Platelet activating factor provokes release of mucin like glycoproteins from guinea pig tracheal epithelial cells via a lipoxygenase-dependent mechanism.}, volume={143}, journal={American Review of Respiratory Disease}, author={Glasgow, W. and Akley, N. J. and Adler, K. B.}, year={1991}, pages={A147} }
 @article{sannes_peters_adler_1991, title={Specific interactions with extracellular matrix may influence epithelial repair mechanisms in the pulmonary alveolus.}, volume={99}, journal={Chest}, author={Sannes, P. and Peters, B. and Adler, K.}, year={1991}, pages={70S–71} }
 @article{low_leslie_hemenway_absher_alder_giancola_vacek_1990, title={Alveolar Type II cell response in rats exposed to aerosols of alpha cristobalite.}, volume={136}, journal={American Journal of Pathology}, author={Low, R. B. and Leslie, K. O. and Hemenway, D. R. and Absher, P. M. and Alder, K. B. and Giancola, M. S. and Vacek, P. M.}, year={1990}, pages={923–931} }
 @article{adler_cheng_kim_1990, title={CHARACTERIZATION OF GUINEA-PIG TRACHEAL EPITHELIAL-CELLS MAINTAINED IN BIPHASIC ORGANOTYPIC CULTURE - CELLULAR COMPOSITION AND BIOCHEMICAL-ANALYSIS OF RELEASED GLYCOCONJUGATES}, volume={2}, ISSN={["1044-1549"]}, DOI={10.1165/ajrcmb/2.2.145}, abstractNote={An air-liquid interface (biphasic) primary culture system in which guinea pig tracheal epithelial cells maintain morphologic characteristics of differentiated epithelium has been developed in this laboratory. In this report, we compared quantitatively cell populations of 8-day cultures to those of epithelial mucosa in intact trachea. In addition, high molecular weight glycoconjugates released by the cultured cells were isolated and characterized. Quantitative morphometric analysis revealed similar volume densities of ciliated, secretory, basal, and "other" cells in cultures and in intact tracheal surface epithelium, although the cultures tended to have smaller cells and contained fewer basal cells. High molecular weight glycoconjugates released apically by cell cultures and excluded from Sepharose CL-4B columns contained approximately 5% hyaluronic acid but undetectable amounts of other proteoglycans, such as chondroitin sulfate, heparan sulfate, and dermatan sulfate. The hyaluronidase-resistant glycoconjugates exhibited a peak buoyant density at 1.49 g/ml on cesium chloride density gradient centrifugation and were shown to contain mucin-type carbohydrate to peptide linkages (i.e., GalNAc to ser/thr) and an amino acid composition typical of respiratory mucins. The results indicate that this organotypic cell culture system mimics quite closely morphology of mucosal epithelium in intact airways and that the cells release high molecular weight glycoconjugates with biochemical properties of mucin-type glycoproteins. Thus, this in vitro system appears well-suited for studies of mucin secretion and other functions of respiratory epithelial cells.}, number={2}, journal={AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY}, author={ADLER, KB and CHENG, PW and KIM, KC}, year={1990}, month={Feb}, pages={145–154} }
 @inbook{adler_low_leslie_mitchell_evans_1990, title={Contractile cells in normal and fibrotic lungs.}, DOI={10.1007/978-1-4612-0485-5_3}, abstractNote={Interstitial pulmonary fibrosis represents a group of fibrosing lung diseases, often of unknown etiology, characterized by hypercellularity and deposition of connective tissue within the interstitium of the alveolar wall (32, 65). Increased “stiffness” of the lung leads to reduced lung volumes and lowered dynamic compliance (80). In interstitial fibrosis, as well as several other forms of lung injury, there is an apparent increase in smooth muscle (SM) cells organized into bundles within areas of the lung where these cells normally are present but difficult to detect, such as the alveolar duct (69, 89, 119). This increase in acinar SM can be quite prominent, and, in some cases, the degree of parenchymal SM hyperplasia is sufficiently excessive to be referred to as “muscular cirrhosis of the lung” (2, 7, 33, 110, 152).}, booktitle={Pathology reviews.}, author={Adler, K. B. and Low, R. B. and Leslie, K. O. and Mitchell, J. and Evans, J. N.}, year={1990}, pages={25–37} }
 @article{mossman_marsh_sesko_hill_shatos_doherty_petruska_adler_hemenway_mickey_et al._1990, title={INHIBITION OF LUNG INJURY, INFLAMMATION, AND INTERSTITIAL PULMONARY FIBROSIS BY POLYETHYLENE GLYCOL-CONJUGATED CATALASE IN A RAPID INHALATION MODEL OF ASBESTOSIS}, volume={141}, ISSN={["0003-0805"]}, DOI={10.1164/ajrccm/141.5_Pt_1.1266}, abstractNote={Several in vitro studies suggest the involvement of active oxygen metabolites in cell damage caused by asbestos. To determine if lung injury, inflammation, and asbestosis could be inhibited in vivo in a rapid-onset, inhalation model of disease, a novel method of chronic administration of antioxidant enzymes was developed. In brief, Fischer 344 rats were treated with polyethylene glycol-conjugated (PEG-) superoxide dismutase or catalase in osmotic pumps over a 10-day (5 days/wk for 2 wk) or 20-day (5 days/wk for 2 wk) period of exposure to crocidolite asbestos. Control rats included sham-exposed animals and those exposed to asbestos but receiving chemically inactivated enzymes. After 10 days of exposure to asbestos, lactic dehydrogenase (LDH), alkaline phosphatase, and total protein in bronchoalveolar lavage (BAL) were measured in one group of rats. Total and differnetial cell counts in BAL also were assessed. After 20 days of exposure, lungs of an additional group of rats were evaluated by histopathology and by measurement of hydroxyproline. Asbestos-associated elevations in LDH, protein, and total cell numbers in BAL were reduced in rats receiving PEG-catalase. Decreases in numbers of alveolar macrophages, polymorphonuclear leukocytes, and lymphocytes occurred in these animals. Exposure to asbestos for 20 days caused significant increases in both the amount of hydroxyproline in lung and the severity and extent of fibrotic lesions as determined by histopathology. These indicators of asbestosis were inhibited in a dosage-dependent fashion in rats receiving PEG-catalase. Use of inactivated PEG-catalase failed to boost serum levels of catalase and did not inhibit asbestos-induced elevation of hydroxyproline in lung.(ABSTRACT TRUNCATED AT 250 WORDS)}, number={5}, journal={AMERICAN REVIEW OF RESPIRATORY DISEASE}, author={MOSSMAN, BT and MARSH, JP and SESKO, A and HILL, S and SHATOS, MA and DOHERTY, J and PETRUSKA, J and ADLER, KB and HEMENWAY, D and MICKEY, R and et al.}, year={1990}, month={May}, pages={1266–1271} }
 @article{adler_peters_krunkosky_sannes_1990, title={Interactions between extracellular matrix, airway and alveolar epithelium: alterations in fibrosis and possible pathogenetic significance.}, volume={6}, journal={Proceedings of the Sixth International Colloquium Pulmonary Fibrosis.}, author={Adler, K. B. and Peters, B. P. and Krunkosky, T. and Sannes, P. L.}, year={1990}, pages={39} }
 @article{adler_holdenstauffer_repine_1990, title={OXYGEN METABOLITES STIMULATE RELEASE OF HIGH-MOLECULAR-WEIGHT GLYCOCONJUGATES BY CELL AND ORGAN-CULTURES OF RODENT RESPIRATORY EPITHELIUM VIA AN ARACHIDONIC ACID-DEPENDENT MECHANISM}, volume={85}, ISSN={["0021-9738"]}, DOI={10.1172/jci114436}, abstractNote={Several common pulmonary disorders characterized by mucus hypersecretion and airway obstruction may relate to increased levels of inhaled or endogenously generated oxidants (O2 metabolites) in the respiratory tract. We found that O2 metabolites stimulated release of high-molecular-weight glycoconjugates (HMG) by respiratory epithelial cells in vitro through a mechanism involving cyclooxygenase metabolism of arachidonic acid. Noncytolytic concentrations of chemically generated O2 metabolites (purine + xanthine oxidase) stimulated HMG release by cell and explant cultures of rodent airway epithelium, an effect which is inhibitable by coaddition of specific O2 metabolite scavengers or inhibitors of arachidonic acid metabolism. Addition of O2 metabolites to epithelial cells provoked production of PGF2a, an effect also inhibitable by coaddition of O2 metabolite scavengers or inhibitors of arachidonic acid metabolism. Finally, addition of exogenous PGF2a to cell cultures stimulated HMG release. We conclude that O2 metabolites increase release of respiratory HMG through a mechanism involving cyclooxygenase metabolism of arachidonic acid with production mainly of PGF2a. This mechanism may be fundamental to the pathogenesis of a variety of lung diseases associated with hypersecretion of mucus and/or other epithelial fluids, as well as a basic cellular response to increased oxidants.}, number={1}, journal={JOURNAL OF CLINICAL INVESTIGATION}, author={ADLER, KB and HOLDENSTAUFFER, WJ and REPINE, JE}, year={1990}, month={Jan}, pages={75–85} }
 @article{adler_akley_lee_1990, title={Oxygen metabolites stimulate inositol lipid turnover in guinea pig airway epithelial cells in organotypic culture.}, volume={141}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Akley, N. J. and Lee, J.}, year={1990}, pages={A107} }
 @article{sannes_peters_adler_1990, title={Specific interactions with extracellular matrix may influence epithelial repair mechanisms in the pulmonary alveolus.}, volume={30}, journal={Proceedings of the 30th Aspen Lung Conference.}, author={Sannes, P. and Peters, B. and Adler, K.}, year={1990}, pages={24} }
 @inbook{adler_akley_ayers_holden-stauffer_khosla_kim_1989, title={A novel culture system for maintaining differentiated epithelial cells between air and liquid phases: A possible model for studying interactions between mineral fibers, gases and airway epithelium.}, booktitle={Biological interaction of inhaled mineral fibers and cigarette smoke}, author={Adler, K. B. and Akley, N. J. and Ayers, G. and Holden-Stauffer, W. J. and Khosla, J. B. and Kim, K. C.}, year={1989}, pages={381–390} }
 @article{mitchell_woodcock-mitchell_reynolds_low_leslie_adler_gabbiani_skalli_1989, title={Alpha smooth muscle actin in parenchymal cells of bleomycin injured rat lung.}, volume={60}, journal={Laboratory Investigation}, author={Mitchell, J. and Woodcock-Mitchell, J. and Reynolds, S. and Low, R. and Leslie, K. and Adler, K. B. and Gabbiani, G. and Skalli, O.}, year={1989}, pages={643–650} }
 @article{adler_low_leslie_mitchell_evans_1989, title={Biology of disease: Contractile cells in normal and fibrotic lungs.}, volume={60}, journal={Laboratory Investigation}, author={Adler, K. B. and Low, R. B. and Leslie, K. O. and Mitchell, J. and Evans, J. N.}, year={1989}, pages={473–485} }
 @article{adler_akley_holden-stauffer_1989, title={Oxidant injury to rodent airway epithelium in primary culture stimulates intracellular production of oxygen free radicals, arachidonic acid metabolism, and mucin secretion.}, volume={139}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Akley, N. J. and Holden-Stauffer, W. J.}, year={1989}, pages={A403} }
 @article{whitcutt_adler_wu_1988, title={A biphasic chamber system for maintaining polarity of differentiation of cultured respiratory tract epithelial cells.}, volume={24}, DOI={10.1007/bf02628493}, abstractNote={A simple, disposable, biphasic cultivation chamber has been developed for respiratory tract epithelial cells. This chamber, the Whitcutt chamber, contains a movable, transparent, permeable gelatin membrane that can be employed either submerged in the culture medium, thereby feeding the cells by the traditional immersion method, or raised to the surface of the culture medium, to bring the apical surfaces of the cells into contact with air and provide nutrients only from below (basal feeding). The effects of biphasic cultivation on the growth and differentiation of respiratory tract epithelial cells from different sources have been studied in Whitcutt chambers. Primary hamster tracheal epithelial (HTE) cells grown to confluence with basal feeding developed a ciliated columnar morphology, with differentiated features (cilia and mucous granules) located in the apical region of the epithelial layer. These cells secreted mucinlike molecules from the apical surface (i.e. the surface in contact with air). Although the apical localization of differentiation features was greater, mucous cell differentiation achieved by basal feeding was quantitatively not greater than that achieved by continuous immersion feeding. Similarly, basal feeding did not alter the degree of epithelial cell differentiation in cultures derived from rat, rabbit, and monkey tracheas or from human bronchial and nasal tissues. In contrast, the differentiation of guinea pig tracheal epithelial cells in culture was significantly influenced by the feeding method employed. When fed basally, guinea pig tracheal epithelial cell cultures expressed various mucociliary functions with resemblance to mucociliary layers in vivo, whereas constantly immersed cultures seemed stratified and squamous. These results suggest that, at least for guinea pigs, the combination of feeding methods provided by the Whitcutt chamber can be used to achieve differentiated cultures of tracheal epithelial cells with a polarity of differentiation that is similar to that observed in intact airways in vivo.}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Whitcutt, J. M. and Adler, K. B. and Wu, R.}, year={1988}, pages={420–428} }
 @article{adler_kim_1988, title={A new culture system for maintaining differentiated rodent respiratory epithelial cells between air and liquid phases: an excellent model for studying interactions between mineral fibers and airway epithelium in vitro.}, volume={4}, journal={Proceedings of the Fourth International Workshop on Effects on Cells (Quebec, 1988)}, author={Adler, K. B. and Kim, K. C.}, year={1988}, pages={10} }
 @article{coflesky_adler_evans_1988, title={Alterations in pulmonary vascular responsiveness following hyperoxic injury to the lung.}, volume={93}, journal={Chest}, author={Coflesky, J. T. and Adler, K. B. and Evans, J. N.}, year={1988}, pages={147S–149} }
 @article{coflesky_adler_woodcock-mitchell_mitchell_evans_1988, title={An autoradiographic and biochemical analysis of cellular proliferation in pulmonary arteries following in vivo hyperoxia.}, volume={137}, journal={American Review of Respiratory Disease}, author={Coflesky, J. T. and Adler, K. B. and Woodcock-Mitchell, J. and Mitchell, J. and Evans, J. N.}, year={1988}, pages={388} }
 @article{low_adler_woodcockmitchell_giancola_vacek_1988, title={BRONCHOALVEOLAR LAVAGE LIPIDS DURING DEVELOPMENT OF BLEOMYCIN-INDUCED FIBROSIS IN RATS - RELATIONSHIP TO ALTERED EPITHELIAL-CELL MORPHOLOGY}, volume={138}, ISSN={["0003-0805"]}, DOI={10.1164/ajrccm/138.3.709}, abstractNote={Alterations in the structure and function of alveolar epithelial cells may contribute to the interstitial fibrosis that can develop following lung injury. The present studies were undertaken to determine if alterations observed in alveolar epithelial cell morphology and cytoskeletal composition are reflected in the profile of bronchoalveolar lavage (BAL) lipids recovered from injured lung. BAL protein and lipid analyses were performed on fluids recovered from control rats and from rats 7, 14, and 28 days after intratracheal instillation of bleomycin, an antineoplastic agent well-known to cause pulmonary interstitial fibrosis. There were increases in recovery of total protein, nonpolar lipid, polar lipid, and phospholipid following bleomycin treatment. The recovery of saturated phosphatidylcholine was increased, but recovery of a second surfactant phospholipid, phosphatidylglycerol, was unchanged, resulting in a significant change in their ratio. The recoveries of cholesterol, cholesterol ester, and triglyceride also were elevated. Changes in the proportional recoveries of neutral lipids, such as cholesterol and saturated phospholipids, could partly explain concurrent reductions in lung compliance that have been described. Changes in lavage lipids paralleled both the process of alveolar reepithelialization and altered expression of alveolar epithelial cell cytoskeletal proteins. Changes in lipid metabolism by alveolar epithelial cells following bleomycin-induced lung injury may be responsible for altered lavage lipid recovery and may directly be related to processes that take place during alveolar type II cell hyperplasia followed by transition to type I cells. BAL lipid analyses thus may provide a relatively noninvasive way of assessing these events.}, number={3}, journal={AMERICAN REVIEW OF RESPIRATORY DISEASE}, author={LOW, RB and ADLER, KB and WOODCOCKMITCHELL, J and GIANCOLA, MS and VACEK, PM}, year={1988}, month={Sep}, pages={709–713} }
 @article{adler_holden-stauffer_1988, title={Exposure of rodent airway epithelium in vitro to oxidants stimulates cyclooxygenase metabolism of arachidonic acid.}, volume={5}, journal={Proceedings of the 5th International Colloquium on Pulmonary Fibrosis (Lyon, France, 1988)}, author={Adler, K. B. and Holden-Stauffer, W. J.}, year={1988}, pages={5} }
 @article{adler_khosla_kim_1988, title={Morphometric and biochemical characterization of a chamber system for maintaining differentiated guinea pig respiratory mucosal cells between air and liquid phases.}, volume={137}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Khosla, J. and Kim, K. C.}, year={1988}, pages={14} }
 @article{adler_akley_holden-stauffer_1988, title={Oxidant injury to airway epithelium in vitro stimulates cyclooxygenase metabolism of arachidonic acid and secretion of mucin.}, volume={107}, journal={Journal of Cell Biology}, author={Adler, K. B. and Akley, N. J. and Holden-Stauffer, W. J.}, year={1988}, pages={350a} }
 @inbook{adler_akley_1988, title={Oxygen radicals stimulate secretion of mucin by rodent airway epithelial cells in organotypic culture.}, booktitle={Oxy radicals in molecular biology and pathology}, author={Adler, K. B. and Akley, N. J.}, year={1988}, pages={101–108} }
 @article{adler_1988, title={Oxygen radicals stimulate secretion of mucin by rodent airway epithelial cells in organotypic culture.}, volume={12A}, journal={Journal of Cellular Biochemistry}, author={Adler, K. B.}, year={1988}, pages={47} }
 @article{coflesky_adler_woodcock mitchell_mitchell_evans_1988, title={Proliferative changes in the pulmonary arterial wall during short term hyperoxic injury to the lung.}, volume={132}, journal={American Journal of Pathology}, author={Coflesky, J. M. and Adler, K. B. and Woodcock Mitchell, J. and Mitchell, J. and Evans, J. N.}, year={1988}, pages={563–573} }
 @article{brody_bitterman_adler_rannels_thet l. a._i._1988, title={The lung matrix and inflammation: Part II. Biochemical and molecular mechanisms of fibrogenesis: implications for environmental lung disease}, volume={138}, journal={American Review of Respiratory Disease}, author={Brody, A. R. and Bitterman, P. B. and Adler, K. B. and Rannels, D. E. and Thet L. A., Rom W. N. and I., Rennard S.}, year={1988}, pages={1056–1057} }
 @article{adler_schwarz_whitcutt_wu_1987, title={A new chamber system for maintaining differentiated guinea pig respiratory epithelial cells between air and liquid phases.}, volume={5}, journal={Biotechniques}, author={Adler, K. B. and Schwarz, J. E. and Whitcutt, J. M. and Wu, R.}, year={1987}, pages={462–467} }
 @article{leslie_woodcock-mitchell_mitchell_adler_low_skalli_gabbiani_1987, title={Alpha smooth muscle actin distribution in developing and adult lung.}, volume={105}, journal={Journal of Cell Biology}, author={Leslie, K. O. and Woodcock-Mitchell, J. and Mitchell, J. and Adler, K. and Low, R. and Skalli, O. and Gabbiani, G.}, year={1987}, pages={25a} }
 @article{coflesky_adler_evans_1987, title={Alterations in pulmonary vascular responsiveness following hyperoxic injury to the lung.}, journal={Proceedings of the 27th Aspen Lung Conference.}, author={Coflesky, J. T. and Adler, K. B. and Evans, J. N.}, year={1987}, pages={18} }
 @article{mossman_marsh_gilbert r._d. h._a._a._s._m. a._j._k. b._et al._1987, title={Dosage dependent inhibition of lung injury, inflammation and fibrosis by polyethylene glycol (PEG)-conjugated catalase in rats exposed by inhalation to asbestos.}, volume={135}, journal={American Review of Respiratory Disease}, author={Mossman, B. T. and Marsh, J. P. and Gilbert R., Hardwick and D. H., Sesko and A., Weller and A., Hill and S., Shatos and M. A., Doherty and J., Adler and K. B., Hemenway and et al.}, year={1987}, pages={A165} }
 @article{butler_adler_evans_szarek_1987, title={Modulation of rabbit airway smooth muscle responsiveness by respiratory epithelium: involvement of an inhibitory metabolite of arachidonic acid.}, volume={135}, journal={American Review of Respiratory Disease}, author={Butler, G. B. and Adler, K. B. and Evans, J. N. and Szarek, J.}, year={1987}, pages={1099–1104} }
 @article{adler_schwarz_anderson_welton_1987, title={PLATELET-ACTIVATING-FACTOR STIMULATES SECRETION OF MUCIN BY EXPLANTS OF RODENT AIRWAYS IN ORGAN-CULTURE}, volume={13}, ISSN={["0190-2148"]}, DOI={10.3109/01902148709064307}, abstractNote={Platelet activating factor (PAF; 1-o-alkyl-2-0-acetyl-sn-glycero-3-phosphocholine-3-phosphocholine) a potential mediator of anaphylaxis, stimulates secretion of mucin by explants of trachea from four separate rodent species (guinea pig, rat, rabbit, ferret) in organ culture. Enhanced secretion is not a result of cell damage or release of histamine by cells within the explants (e.g., platelets). It is inhibited by equimolar concentrations of the potent PAF-receptor antagonist, Ro 19-3704. PAF provokes production of immunoreactive peptidyl leukotrienes (ir-LTC4, LTD4, LTE4) within the explants. The stimulatory effect of PAF on mucin secretion is blocked by equimolar concentrations of nordihydroguiaretic acid (NDGA) a "mixed" inhibitor of both cyclo- and lipoxygenase pathways of arachidonic acid metabolism. Leukotrienes are localized within tracheobronchial epithelium by immunohistochemical staining, and physical removal of epithelium from explants inhibits production of leukotrienes in vitro under nonstimulated conditions and after exposure to PAF. In addition, the stimulatory effect of PAF on mucin secretion is not altered by FPL-55712, a receptor antagonist of LTD4. These results are consistent with the hypothesis that PAF stimulates secretion of mucin by activating biosynthesis of lipoxygenase products (e.g., peptidyl leukotrienes) within epithelial cells of the respiratory mucosa.}, number={1}, journal={EXPERIMENTAL LUNG RESEARCH}, author={ADLER, KB and SCHWARZ, JE and ANDERSON, WH and WELTON, AF}, year={1987}, pages={25–43} }
 @article{adler_repine_1987, title={Superoxide anion stimulates arachidonic acid dependent secretion of mucin by explant and cell cultures of rodent respiratory epithelium.}, volume={135}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Repine, J. R.}, year={1987}, pages={A164} }
 @article{szarek_buter_adler_evans_1986, title={A cyclooxygenase product of arachidonic acid is involved in epithelium mediated modulation of rabbit airway smooth muscle responsiveness.}, volume={133}, journal={American Review of Respiratory Disease}, author={Szarek, J. L. and Buter, G. B. and Adler, K. B. and Evans, J. N.}, year={1986}, pages={A115} }
 @article{adler_whitcutt_wu_1986, title={A new chamber system for maintaining differentiated guinea pig respiratory epithelial cells between air and liquid phases.}, volume={103}, journal={Journal of Cell Biology}, author={Adler, K. B. and Whitcutt, J. M. and Wu, R.}, year={1986}, pages={207a} }
 @article{adler_callahan_evans_1986, title={Alterations in the cellular population of the alveolar wall in  bleomycin induced pulmonary fibrosis in rats: an ultrastructural morphometric study.}, volume={133}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Callahan, L. M. and Evans, J. N.}, year={1986}, pages={1043–1048} }
 @article{callahan_evans_adler_1986, title={Alterations in the cellular population of the alveolar wall in an animal model of fibrosis: a morphometric study.}, volume={89}, journal={Chest}, author={Callahan, L. M. and Evans, J. N. and Adler, K. B.}, year={1986}, pages={189S–190} }
 @article{adler_hendley_davis_1986, title={Bacteria associated with chronic obstructive pulmonary disease elaborate extracellular products that stimulate secretion of mucin by explants of guinea pig airways.}, volume={125}, journal={American Journal of Pathology}, author={Adler, K. B. and Hendley, D. D. and Davis, G. S.}, year={1986}, pages={501–514} }
 @article{coflesky_adler_anderson_evans_1986, title={Effects of hyperoxia on metabolism of arachidonic acid by pulmonary vessels and airways.}, volume={22}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Coflesky, J. T. and Adler, K. B. and Anderson, W. H. and Evans, J. N.}, year={1986}, pages={44A} }
 @inbook{brody_hill_hesterberg_barrett_adler_1986, title={Intracellular translocation of inorganic particles.}, DOI={10.1007/978-1-4613-2161-3_17}, booktitle={The cytoskeleton.}, author={Brody, A. R. and Hill, L. H. and Hesterberg, T. W. and Barrett, J. C. and Adler, K. B.}, year={1986}, pages={221–227} }
 @article{mossman_adler_1986, title={Mechanisms of asbestos induced toxicity and lung disease.}, volume={22}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Mossman, B. T. and Adler, K. B.}, year={1986}, pages={49A} }
 @inbook{adler_1986, title={Mucin secretion by respiratory tract tissue in vitro.}, booktitle={In vitro models of respiratory epithelium}, author={Adler, K. B.}, year={1986}, pages={27–50} }
 @inbook{adler_schwarz_repine_1986, title={Oxygen free radicals stimulate secretion of mucin by rodent respiratory epithelium in vitro.}, booktitle={Microbeam analysis -- 1986.}, author={Adler, K. B. and Schwarz, J. E. and Repine, J. E.}, year={1986}, pages={571–573} }
 @article{adler_schwarz_anderson_welton_1986, title={Platelet activating factor stimulates secretion of mucin by explants of rodent airways in organ culture.}, volume={133}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Schwarz, J. E. and Anderson, W. H. and Welton, A. F.}, year={1986}, pages={A212} }
 @article{adler_callahan_evans_1985, title={Alterations in the cellular population of the alveolar wall in an animal model of fibrosis: a morphometric study.}, journal={Proceedings of the Aspen Lung Conference.}, author={Adler, K. B. and Callahan, L. M. and Evans, J. N.}, year={1985}, pages={24} }
 @article{adler_schwartz_butler_1985, title={Effects of arachidonate metabolites on mucin secretion by explants of rabbit bile and pancreatic ducts in organ culture.}, volume={26}, journal={404nOtfound}, author={Adler, K. B. and Schwartz, J. E. and Butler, G. B.}, year={1985}, pages={47} }
 @inbook{adler_butler_hemenway_schwarz_banks_evans_1985, title={Exposure of small airways to cristobalite in vitro.}, volume={G-3}, booktitle={In vitro effects of mineral dusts.}, author={Adler, K. B. and Butler, G. B. and Hemenway, D. R. and Schwarz, J. E. and Banks, P. O. and Evans, J. N.}, year={1985}, pages={293–301} }
 @article{shatos_adler_doherty_orfeo_mossman_1985, title={Localization of intracellular and extracellular SOD in cells of the lung:  increased enzyme activity after exposure to asbestos.}, volume={131}, journal={American Review of Respiratory Disease}, author={Shatos, M. A. and Adler, K. B. and Doherty, J. M. and Orfeo, T. and Mossman, B. T.}, year={1985}, pages={A187} }
 @article{butler_adler_evans_szarek_1985, title={Modulation of rabbit airway smooth muscle tone by the respiratory epithelium is mediated by a dilatory prostaglandin.}, volume={27}, journal={404nOtfound}, author={Butler, G. B. and Adler, K. B. and Evans, J. N. and Szarek, J. L.}, year={1985}, pages={187} }
 @article{adler_callahan_evans_1985, title={Quantitative measure of cellular changes in pulmonary parenchyma of rats with bleomycin induced fibrosis.}, volume={131}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Callahan, L. M. and Evans, J. N.}, year={1985}, pages={A32} }
 @article{adler_schwarz_repine_1985, title={Superoxide, but not peroxide, stimulates secretion of mucin by explants of rodent airway tissue in organ culture.}, volume={101}, journal={Journal of Cell Biology}, author={Adler, K. B. and Schwarz, J. E. and Repine, J. E.}, year={1985}, pages={104a} }
 @article{low_mitchell_evans_adler_1984, title={Actin content in normal and bleomycin fibrotic rat lung.}, volume={129}, journal={American Review of Respiratory Disease}, author={Low, R. B. and Mitchell, J. W. and Evans, J. N. and Adler, K. B.}, year={1984}, pages={311–316} }
 @article{adler_butler_welton_1984, title={Cyclooxygenase and lipoxygenase metabolites of arachidonic acid influence mucin secretion by explants of rodent airways.}, volume={99}, journal={Journal of Cell Biology}, author={Adler, K. B. and Butler, G. B. and Welton, A. F.}, year={1984}, pages={929} }
 @article{adler_hendley_schwarz_davis_1984, title={Exoproducts from bacteria associated with chronic bronchitis stimulate secretion of mucin by rodent tracheal explants in organ culture.}, volume={129}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Hendley, D. D. and Schwarz, J. E. and Davis, G. S.}, year={1984}, pages={A182} }
 @article{adler_mossman_butler_jean_craighead_1984, title={INTERACTION OF MOUNT ST-HELENS VOLCANIC ASH WITH CELLS OF THE RESPIRATORY EPITHELIUM}, volume={35}, ISSN={["0013-9351"]}, DOI={10.1016/0013-9351(84)90142-7}, abstractNote={Respirable-sized dust from the Mount St. Helens (MSH) eruption of Spring 1980, and minerals similar to the major components of the volcanic ash, were examined comparatively for interactions with epithelial cells of rodent respiratory airways in vitro. MSH dust, Na feldspar, cristobalite, and alpha-quartz, in concentrations of 0.4 to 40 mg/ml, had neither significant effects on mucin release by tracheal explants nor acute toxic effects after exposure for 2 hr. Long-term incubation (1 and 3 weeks) of explants after a 1-hr exposure to MSH dust failed to elicit widespread toxic or proliferative changes in airway epithelial cells. In contrast, long-term exposure to Na feldspar, cristobalite, and alpha-quartz caused significant toxicity to the explants, although metaplastic changes were not observed. Ultrastructural evidence of associations (e.g., phagocytosis) between particulates and respiratory epithelium was not observed. The results of these studies suggest that volcanic ash from MSH interacts minimally with cells of the respiratory mucosa.}, number={2}, journal={ENVIRONMENTAL RESEARCH}, author={ADLER, KB and MOSSMAN, BT and BUTLER, GB and JEAN, LM and CRAIGHEAD, JE}, year={1984}, pages={346–361} }
 @article{woodcock-mitchell_adler_low_1984, title={Immunohistochemical identification of cell types in normal and in bleomycin induced fibrotic rat lung.}, volume={130}, journal={American Review of Respiratory Disease}, author={Woodcock-Mitchell, J. and Adler, K. B. and Low, R. B.}, year={1984}, pages={910–916} }
 @article{adler_butler_evans_1984, title={Involvement of prostaglandins in both mucin secretion and contractility of rabbit pancreatic and bile ducts in vitro.}, volume={25}, journal={404nOtfound}, author={Adler, K. B. and Butler, G. B. and Evans, J. N.}, year={1984}, pages={195} }
 @article{adler_hendley_1984, title={Stimulation of airway mucin secretion by an exoproduct elaborated by Pseudomonas aeruginosa: preliminary isolation.}, volume={25}, journal={404nOtfound}, author={Adler, K. B. and Hendley, D. D.}, year={1984}, pages={121} }
 @article{brody_hill_stirewalt_adler_1983, title={Actin containing microfilaments of pulmonary epithelial cells provide a mechanism for translocating inhaled asbestos to the interstitium.}, volume={83}, journal={Chest}, author={Brody, A. R. and Hill, L. and Stirewalt, W. and Adler, K. B.}, year={1983}, pages={11S–13} }
 @article{evans_krill_adler_low_kelley_1983, title={Active cellular control of alveolar compliance.}, volume={13}, journal={Current Problems in Clinical Biochemistry}, author={Evans, J. N. and Krill, J. and Adler, K. B. and Low, R. B. and Kelley, J.}, year={1983}, pages={142–148} }
 @article{craighead_adler_butler_emerson_mossman_woodworth_1983, title={Biology of disease: health effects of Mount St. Helens volcanic dust.}, volume={48}, journal={Laboratory Investigation}, author={Craighead, J. E. and Adler, K. B. and Butler, G. B. and Emerson, R. J. and Mossman, B. T. and Woodworth, C. W.}, year={1983}, pages={5–12} }
 @article{adler_butler_1983, title={Cellular mechanisms of mucin secretion by rodent epithelial cells in vitro.}, volume={97}, journal={Journal of Cell Biology}, author={Adler, K. B. and Butler, G. B.}, year={1983}, pages={437a} }
 @article{low_mitchell_evans_adler_1983, title={Contractile proteins of the lung.}, volume={13}, journal={Current Problems in Clinical Biochemistry}, author={Low, R. B. and Mitchell, J. W. and Evans, J. N. and Adler, K. B.}, year={1983}, pages={149–156} }
 @article{adler_krill_alberghini_evans_1983, title={EFFECT OF CYTOCHALASIN-D ON SMOOTH-MUSCLE CONTRACTION}, volume={3}, ISSN={["0886-1544"]}, DOI={10.1002/cm.970030521}, abstractNote={Abstract}, number={5-6}, journal={CELL MOTILITY AND THE CYTOSKELETON}, author={ADLER, KB and KRILL, J and ALBERGHINI, TV and EVANS, JN}, year={1983}, pages={545–551} }
 @article{adler_krill_alberghini_evans_1983, title={Effects of cytochalasin D on contraction of isolated pulmonary vessels.}, volume={2}, journal={Proceedings of the 2nd International Conference on Cell Biol Struct.}, author={Adler, K. B. and Krill, J. and Alberghini, T. and Evans, J. N.}, year={1983}, pages={15} }
 @article{woodcock-mitchell_adler_low_1983, title={Immunohistochemical studies of cell types in normal and fibrotic rat lung.}, volume={127}, journal={American Review of Respiratory Disease}, author={Woodcock-Mitchell, J. and Adler, K. B. and Low, R. B.}, year={1983}, pages={68} }
 @article{adler_alberghini_counts_auletta_1983, title={SECRETION OF MUCIN BY EXPLANTS OF RABBIT AND HUMAN CERVIX IN ORGAN-CULTURE}, volume={29}, ISSN={["0006-3363"]}, DOI={10.1095/biolreprod29.3.751}, abstractNote={Small explants (2-3 mm3) of endocervix from virgin, estrous rabbits, and from hospitalized patients undergoing hysterectomy for nonneoplastic disease, were placed in organ culture and maintained in serum-free media for 4 days at 35 degrees C in a humid environment of 95% air/5% CO2. Waymouth's MB 752/1 with 10-5 M hydrocortisone succinate, 10-7 M retinyl acetate, and 1 microgram/ml insulin proved to be an excellent medium for maintaining these tissues, as judged by examination with light and scanning electron microscopy after incubation for 5 days. The explants incorporated the radiolabeled glycoprotein precursor, tritiated glucosamine, and secreted labeled mucin glycoproteins in vitro. Mucin released into the culture medium contained sialic acid and hexosamine in a molar ratio of approximately 0.5-0.8:1.0. Although some alterations occur in the morphology of secretory cells and their products after maintenance in culture for several days, the system can be utilized for studying various aspects of the cell biology of cervical mucin secretion.}, number={3}, journal={BIOLOGY OF REPRODUCTION}, author={ADLER, KB and ALBERGHINI, TV and COUNTS, DF and AULETTA, FJ}, year={1983}, pages={751–765} }
 @article{adler_winn_alberghini_craighead_1983, title={Stimulatory effect of Pseudomonas aeruginosa on mucin secretion by respiratory epithelium.}, volume={249}, journal={404nOtfound}, author={Adler, K. B. and Winn, W.C., Jr. and Alberghini, T. V. and Craighead, J. E.}, year={1983}, pages={1615–1617} }
 @article{evans_low_kelley_j._adler_1983, title={The myofibroblast in pulmonary fibrosis.}, volume={83}, journal={Chest}, author={Evans, J. N. and Low, R. B. and Kelley, J. Krill and J. and Adler, K. B.}, year={1983}, pages={97S–99} }
 @article{adler_winn_hardwick_craighead_1982, title={EFFECTS OF BACTERIAL PRODUCTS ON SECRETION OF MUCIN BY RODENT TRACHEA INVITRO}, volume={81}, ISSN={["0012-3692"]}, DOI={10.1378/chest.81.5_supplement.37s}, abstractNote={The lungs and airways often become either colonized or infected by bacteria in many respiratory diseases characterized by hypersecretion of mucin (cystic fibrosis and chronic bronchitis). Despite the concomitant occurrence of bacterial presence and excessive secretion of mucin, surprisingly little scientific attention has been allotted to studies of possible interactions between bacteria or bacterial products and tracheobronchial mucin-secreting cells. In this report, we describe experiments designed to investigate the effects of specific bacteriaderived substances on secretion of mucin by guinea pig tracheal explants in short term (2 hr) organ cultures.}, number={5}, journal={CHEST}, author={ADLER, KB and WINN, WC and HARDWICK, DH and CRAIGHEAD, JE}, year={1982}, pages={S37–S39} }
 @article{low_evans_adler_1982, title={Effects of bleomycin induced pulmonary fibrosis on lung actin content and polymerization.}, volume={125}, journal={American Review of Respiratory Disease}, author={Low, R. B. and Evans, J. N. and Adler, K. B.}, year={1982}, pages={218} }
 @article{evans_kelley_low_adler_1982, title={Increased contractility of isolated lung parenchyma in an animal model of pulmonary fibrosis induced by bleomycin.}, volume={125}, journal={American Review of Respiratory Disease}, author={Evans, J. N. and Kelley, J. and Low, R. B. and Adler, K. B.}, year={1982}, pages={89–94} }
 @article{mossman_adler_jean_craighead_1982, title={Mechanisms of hypersecretion in rodent tracheal explants after exposure to chrysotile asbestos: studies using lectins.}, volume={91}, journal={Chest}, author={Mossman, B. T. and Adler, K. B. and Jean, L. and Craighead, J. E.}, year={1982}, pages={23S–25} }
 @article{adler_hardwick_craighead_1982, title={PORCINE TRACHEAL GOBLET CELL ULTRASTRUCTURE - A 3-DIMENSIONAL RECONSTRUCTION}, volume={3}, ISSN={["1521-0499"]}, DOI={10.3109/01902148209115817}, abstractNote={In order to study specific anatomical relationships among organelles involved in the movement and secretion of mucin granules by airway epithelium, a three-dimensional replica of a single goblet cell from porcine trachea was constructed by means of transmission electron microscopy of serially prepared ultrathin sections. Many features of the goblet cell that could not be deduced from single planes of section were revealed. The cell is columnar in configuration. The cytoplasm contains not one, but several discrete clusters of mucin granules, each associated with a separate Golgi apparatus. Microtubules and microfilaments often are observed in close association with both mucin granules and coiled filamentous mitochondria, suggesting a role in the intracellular movement of these organelles. These morphological observations are in agreement with previous functional studies of airway mucin secretion, and elucidate further the cellular biology of airway mucosal goblet cells and the secretory process.}, number={1}, journal={EXPERIMENTAL LUNG RESEARCH}, author={ADLER, KB and HARDWICK, DH and CRAIGHEAD, JE}, year={1982}, pages={69–80} }
 @article{adler_craighead_vallyathan_evans_1981, title={Actin containing cells in human pulmonary fibrosis.}, volume={102}, journal={American Journal of Pathology}, author={Adler, K. B. and Craighead, J. E. and Vallyathan, N. V. and Evans, J. N.}, year={1981}, pages={427–438} }
 @article{emerson_adler_evans_davis_1981, title={Alveolar macrophage heterogeneity in silica exposed rats.}, volume={123}, journal={American Review of Respiratory Disease}, author={Emerson, R. J. and Adler, K. B. and Evans, J. N. and Davis, G. S.}, year={1981}, pages={138} }
 @article{adler_hardwick_craighead_1981, title={Effect of cholera toxin on secretion of mucin by explants of guinea pig trachea.}, volume={45}, journal={Laboratory Investigation}, author={Adler, K. B. and Hardwick, D. H. and Craighead, J. E.}, year={1981}, pages={372–377} }
 @article{adler_winn_craighead_1981, title={Effects of bacterial products on secretion of mucin by rodent trachea in vitro.}, volume={24}, journal={Proceedings of the 24th Aspen Lung Conference.}, author={Adler, K. B. and Winn, W. C. and Craighead, J. E.}, year={1981}, pages={21} }
 @article{mossman_adler_craighead_1981, title={Mechanisms of hypersecretion in rodent tracheal explants after exposure to chrysotile asbestos.}, volume={24}, journal={Proceedings of the 24th Aspen Lung Conference.}, author={Mossman, B. T. and Adler, K. B. and Craighead, J.}, year={1981}, pages={13} }
 @article{adler_st. andre-ukena_beeken_1981, title={Preliminary characterization of macrophages isolated from the human colon.}, volume={80}, journal={Gastroenterology}, author={Adler, K. B. and St. Andre-Ukena, S. and Beeken, W.}, year={1981}, pages={1097} }
 @article{adler_winn_hardwick_craighead_1981, title={Stimulation by bacterial products of mucin secretion in rodent tracheal explants.}, volume={22}, journal={404nOtfound}, author={Adler, K. B. and Winn, W. C., Jr. and Hardwick, D. H. and Craighead, J. E.}, year={1981}, pages={91} }
 @article{adler_brody_craighead_1981, title={Studies on the mechanism of mucin secretion by cells of the porcine tracheal epithelium.}, volume={166}, DOI={10.3181/00379727-166-41030}, abstractNote={Abstract The effects of selected pharmacological agents on mucin secretion by the tracheal epithelium of piglets were studied using organ culture. Mucin release into the culture medium was reduced, and mucin retention by secretory cells of the mucosa and submucosal glands was increased, by colchicine, vinblastine sulfate, and cytochalasin B. Dibutyryl cyclic AMP had no significant effect. Although these agents have diverse and poorly understood influences on cell functions, the results suggest a possible role for mi-crotubules and microfilaments in the intracellular movement and release of mucin.}, journal={Proceedings of the Society for Experimental Biology and Medicine}, author={Adler, K. B. and Brody, A. R. and Craighead, J. E.}, year={1981}, pages={96–106} }
 @article{evans_adler_1981, title={The lung strip: evaluation of a method to study contractility of pulmonary parenchyma.}, volume={2}, DOI={10.3109/01902148109052314}, abstractNote={Isolated strips of rabbit lung were examined as an in vitro model for assessment of the direct effect of pharmacologic agents on the pulmonary parenchyma. Changes in force of the strip were measured with an isometric force transducer. Histamine, acetylcholine and epinephrine elicited dose-related contractile responses. Morphological and immunohistochemical examination revealed three possible sources of force generation within the strip: airway smooth muscle, vascular smooth muscle, and interstitial actin-containing cells. Generation of force by the strip could reflect contraction of any combination of these three elements. Therefore, ascription of such contraction to peripheral airway smooth muscle alone is questionable. In order to assess the properties of the contractile elements within the strip, it is necessary to isolate and study them individually.}, journal={Experimental Lung Research}, author={Evans, J. N. and Adler, K. B.}, year={1981}, pages={187–194} }
 @article{adler_craighead_1980, title={Cellular mechanisms of respiratory tract mucin secretion.}, volume={8}, journal={Proceedings of the 8th International Congress on Cystic Fibrosis.}, author={Adler, K. B. and Craighead, J. E.}, year={1980}, pages={1a} }
 @article{davis_brody_adler_1980, title={Changes in the surface morphology of human alveolar macrophages induced by tobacco and marijuana smoking.}, volume={1}, journal={Chest}, author={Davis, G. S. and Brody, A. R. and Adler, K. B.}, year={1980}, pages={281–293} }
 @inbook{mossman_adler_craighead_1980, title={Cytotoxic and proliferative changes in tracheal organ and cell cultures after exposure to mineral dusts.}, booktitle={The in vitro effects of mineral dusts}, author={Mossman, B. T. and Adler, K. B. and Craighead, J. E.}, year={1980}, pages={241–250} }
 @article{mossman_ezerman_adler_craighead_1980, title={Isolation and spontaneous transformation of cloned lines of hamster tracheal epithelial cells.}, volume={40}, journal={Cancer Research}, author={Mossman, B. T. and Ezerman, E. B. and Adler, K. B. and Craighead, J. E.}, year={1980}, pages={4403–4408} }
 @article{davis_adler_1980, title={Morphologic changes induced in human alveolar macrophages by levamisole treatment.}, volume={121}, journal={American Review of Respiratory Disease}, author={Davis, G. S. and Adler, K. B.}, year={1980}, pages={64} }
 @article{adler_kelley_evans_1980, title={Morphological and pharmacological evidence of increased contractility in human and rat fibrotic lungs.}, volume={121}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Kelley, J. and Evans, J. N.}, year={1980}, pages={309} }
 @article{adler_hardwick_craighead_1980, title={Stimulatory effects of cholera toxin on secretion of mucin by tracheal explants in organ culture.}, volume={87}, journal={Journal of Cell Biology}, author={Adler, K. B. and Hardwick, D. H. and Craighead, J. E.}, year={1980}, pages={307a} }
 @article{davis_adler_brody_1980, title={The morphology of alveolar macrophage immunologic activation: a scanning electron microscopic study.}, volume={121}, journal={American Review of Respiratory Disease}, author={Davis, G. S. and Adler, K. B. and Brody, A. R.}, year={1980}, pages={269} }
 @article{kelley_adler_davis_1979, title={Alveolar macrophage function in desquamative interstitial pneumonitis.}, volume={27}, journal={404nOtfound}, author={Kelley, J. and Adler, K. B. and Davis, G. S.}, year={1979}, pages={569A} }
 @article{davis_brody_adler_1979, title={Functional and physiological correlates of human alveolar macrophage cell shape and surface morphology.}, volume={75S}, DOI={10.1016/s0012-3692(15)30441-4}, abstractNote={The pulmonary alveolar macrophage (PAM) participates in the immunologic responses of the lung, and is the resident phagocyte of the alveolar membrane. 1 Green GM Jakab GJ Low RB et al. Defense mechanisms of the respiratory membrane: State of the art. Am Rev Respir Dis. 1977; 115: 479-514 PubMed Google Scholar The PAM has been described as variable in size and shape, and the cell population as heterogeneous, 2 Brain JD Proctor DF Reid LM Respiratory Defense Mechanisms (part II). Marcel Dekker, New York1977: 849-892 Google Scholar but few attempts have been made to define macrophage shape quantitatively, or to relate cell shape to cell physiology and function.}, journal={Chest}, author={Davis, G. S. and Brody, A. R. and Adler, K. B.}, year={1979}, pages={280–282} }
 @article{adler_hardwick_craighead_1979, title={Mechanisms of mucin secretion by porcine tracheal goblet cells.}, volume={119}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Hardwick, D. H. and Craighead, J. E.}, year={1979}, pages={285} }
 @article{adler_vallyathan_kelley_evans_1979, title={Nonmuscle contractile cells in pulmonary fibrosis.}, volume={83}, journal={Journal of Cell Biology}, author={Adler, K. B. and Vallyathan, N. V. and Kelley, J. and Evans, J. N.}, year={1979}, pages={319a} }
 @article{adler_davis_woodworth_brody_1979, title={The human pulmonary alveolar macrophage: two distinct morphological populations.}, volume={3}, journal={Scanning Electron Microscopy}, author={Adler, K. B. and Davis, G. S. and Woodworth, C. W. and Brody, A. R.}, year={1979}, pages={921–928} }
 @inproceedings{adler_brody_craighead_1978, title={Association of microtubules and microfilaments with mucin granules in tracheal goblet cells their possible role in exocytosis.}, volume={2}, booktitle={Proceedings of the 9th International Congress on Electron Microscopy}, author={Adler, K. B. and Brody, A. R. and Craighead, J. E.}, year={1978}, pages={500–501} }
 @article{evans_previti_adler_vallyathan_1978, title={Contractile properties of peripheral lung tissue.}, volume={21}, journal={404nOtfound}, author={Evans, J. N. and Previti, R. A. and Adler, K. B. and Vallyathan, N. V.}, year={1978}, pages={35} }
 @article{mossman_adler_craighead_1978, title={INTERACTION OF CARBON PARTICLES WITH TRACHEAL EPITHELIUM IN ORGAN-CULTURE}, volume={16}, ISSN={["0013-9351"]}, DOI={10.1016/0013-9351(78)90148-2}, abstractNote={Organ cultures of hamster, porcine, and guinea pig trachea were exposed in vitro to different types and concentrations of carbon particles. At intervals thereafter, morphological changes in the mucosa were documented by light, scanning, and transmission electron microscopy. Secretory cells formed prominent polymorphic cytoplasmic protuberances that were partially filled with secretory granules within minutes after the addition of carbon to cultures. These processes were produced by particles of various sizes; their number increased with the concentrations of carbon in the medium. Differentiated cells of the epithelium sloughed after exposure to carbon for several hours. Subsequently, basal cell hyperplasia was observed. Larger particles of carbon (⩾ 15 μm) occasionally produced squamous metaplasia in the epithelium. Smaller particles (0.5 – 1μm) were transported to the submucosa, where they were taken up by mesenchymal cells. These studies show that the mucosa of the trachea responds to carbon particles in a unique, but characteristic, fashion.}, number={1-3}, journal={ENVIRONMENTAL RESEARCH}, author={MOSSMAN, BT and ADLER, KB and CRAIGHEAD, JE}, year={1978}, pages={110–122} }
 @article{adler_brody_davis_1978, title={Localization of actin in surface ruffles of pulmonary alveolar macrophages: Effects of cytochalasin B, colchicine and cyclic AMP.}, volume={79}, journal={Journal of Cell Biology}, author={Adler, K. B. and Brody, A. R. and Davis, G. S.}, year={1978}, pages={276a} }
 @article{adler_brody_craighead_1978, title={Ultrastructural association of microtubules and microfilaments with mucin granules in porcine tracheobronchial tissue in organ culture.}, volume={37}, journal={404nOtfound}, author={Adler, K. B. and Brody, A. R. and Craighead, J. E.}, year={1978}, pages={266} }
 @article{adler_mossman_craighead_1977, title={Interaction of carbon particles with rodent tracheal epithelium in organ culture.}, volume={13}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Adler, K. B. and Mossman, B. T. and Craighead, J. E.}, year={1977}, pages={202} }
 @article{adler_fand_1977, title={The cilioinhibitory effect of phenothiazine in vitro and its antagonism by calcium}, volume={227}, journal={Archives Internationales de Pharmacodynamie et de Therapie}, author={Adler, K. B. and Fand, I.}, year={1977}, pages={309–322} }
 @article{ezerman_adler_craighead_1976, title={Isolation of epithelial cells from hamster tracheal respiratory epithelium.}, volume={12}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Ezerman, E. B. and Adler, K. B. and Craighead, J. E.}, year={1976}, pages={319} }
 @article{adler_dulfano_1976, title={The rheological factor in mucociliary clearance.}, volume={88}, journal={Journal of Laboratory and Clinical Medicine}, author={Adler, K. B. and Dulfano, M. J.}, year={1976}, pages={22–30} }
 @article{dulfano_adler_1975, title={Physical properties of sputum. VII. Rheologic properties and mucociliary transport.}, volume={112}, journal={American Review of Respiratory Disease}, author={Dulfano, M. J. and Adler, K. B.}, year={1975}, pages={341–348} }
 @article{fand_adler_1974, title={Cilioinhibitory effect of chlorpromazine in vitro and its antagonism by calcium.}, volume={10}, journal={In Vitro Cellular & Developmental Biology. Animal}, author={Fand, I. and Adler, K. B.}, year={1974}, pages={341} }
 @article{dulfano_adler_1974, title={Mucociliary clearance and mucus viscoelasticity.}, volume={66}, journal={Chest}, author={Dulfano, M. J. and Adler, K. B.}, year={1974}, pages={323} }
 @article{adler_dulfano_wooten_1974, title={Physical properties of sputum. V.  The effects of time, freezing and thawing on viscoelasticity measurements.}, volume={109}, journal={American Review of Respiratory Disease}, author={Adler, K. B. and Dulfano, M. J. and Wooten, O.}, year={1974}, pages={490–492} }
 @article{adler_wooten_dulfano_1973, title={MAMMALIAN RESPIRATORY MUCOCILIARY CLEARANCE}, volume={27}, ISSN={["0003-9896"]}, DOI={10.1080/00039896.1973.10666403}, abstractNote={An in vivo model was designed to provide information on the major factors affecting mammalian mucociliary clearance. It involves simultaneous quantitation of mucous load, tracheal transport velocity, ciliary beat frequency, and rheological properties of mucus. Values in 23 cats were obtained. The results were remarkably consistent for the group as well as for individual cats that were reexamined. The mucous blanket depth measures usually < 10μ, ciliary beat frequency averages 876±151 beats per minute, and transport rate 10.5±3.7 mm/min. Average mucous viscosity was 248±106 poises at a shear rate of 1 sec-1 and recoverable shear strain (elasticity) 3.1±1.1 units at 100 dynes/sq cm. It is suggested that this model can serve as a sensitive indicator of early deleterious effects of airway pollution or noxious inhalants and pinpoint the initial target area.}, number={6}, journal={ARCHIVES OF ENVIRONMENTAL HEALTH}, author={ADLER, KB and WOOTEN, O and DULFANO, MJ}, year={1973}, pages={364–369} }
 @article{dulfano_adler_wooten_1973, title={Physical properties of sputum. IV. The effects of 100% humidity and water mist.}, volume={107}, journal={American Review of Respiratory Disease}, author={Dulfano, M. J. and Adler, K. B. and Wooten, O.}, year={1973}, pages={130–133} }
 @article{adler. k. b._o._w._e._dulfano_1972, title={Physical properties of sputum. III. Rheological variability and intrinsic relationships.}, volume={106}, journal={American Review of Public Administration}, author={Adler. K. B., Wooten and O., Philipoff and W., Lerner and E. and Dulfano, M. J.}, year={1972}, pages={86–96} }