@article{von furstenberg_larson_2023, title={Creating a More Inclusive Future for Conservation: Reflections from the “Skirmish in Spokane"}, volume={17}, number={4}, journal={Wildlife Professional}, publisher={The Wildlife Society, Wildlife Professional}, author={von Furstenberg, R.J. and Larson, L.R.}, year={2023}, month={Jul}, pages={31–35} }
 @article{von furstenberg_vayer_larson_nils peterson_lee_stevenson_ahlers_anhalt‐depies_bethke_clark_et al._2023, title={Evaluating impacts of R3 workshops for first‐time hunters at universities across the United States}, volume={47}, ISSN={2328-5540 2328-5540}, url={http://dx.doi.org/10.1002/wsb.1482}, DOI={10.1002/wsb.1482}, abstractNote={AbstractDeclines in hunter numbers across the United States make hunter recruitment, retention, and reactivation (R3) a high priority for wildlife management. As wildlife management agencies and nongovernmental organizations seek to reach new audiences, college campuses present a unique opportunity to cultivate nontraditional path hunters. Despite recent proliferation of R3 initiatives, little research has evaluated effects of hunting programs on knowledge, attitudes, and behaviors of new hunters. We designed and implemented Getting Started Outdoors: Hunting 101 workshops specifically targeting college students without previous hunting experience, and we assessed workshop efficacy with a theoretically‐grounded approach to workshop evaluation. Using quantitative and qualitative analysis of surveys conducted before, shortly after, and 12–18 months after workshops, we assessed impacts of R3 efforts at large public universities in 16 different U.S. states. Across all states, 19 workshops attracted 314 total participants, with 255 completing both pre‐ and post‐workshop assessments and 133 completing the follow‐up surveys. Workshops significantly increased participants' confidence in hunting, reduced barriers related to inadequate knowledge and skills, and fostered positive views of hunters and hunting. Immediately after workshops, most participants said they would definitely (50%) or probably (34%) hunt in the future; 82% said they would likely (or very likely) purchase a hunting license. Over one year after the workshops, 34% of workshop participants reported having hunted, and another 45% said they would probably hunt in the future. Overall, workshops attracted a diverse population of potential hunters, increased interest in future hunting, and created hunting advocates. Findings highlight the potentially powerful impact that R3 programs focused on diverse college students can have on the future of hunting across the United States.}, number={3}, journal={Wildlife Society Bulletin}, publisher={Wiley}, author={von Furstenberg, Richard and Vayer, Victoria R. and Larson, Lincoln R. and Nils Peterson, M. and Lee, Kangjae Jerry and Stevenson, Kathryn and Ahlers, Adam A. and Anhalt‐Depies, Christine and Bethke, Taniya and Clark, Brian and et al.}, year={2023}, month={Aug} }
 @inproceedings{von furstenberg_larson_2023, title={Mutualism is here; is our conservation system ready?}, booktitle={Pathways 2023: Human Dimensions of Wildlife Conference}, author={von Furstenberg, R. and Larson, L.}, year={2023} }
 @inproceedings{von furstenberg_larson_rutledge_2023, title={Using social science to inform and advance R3 programming in North Carolina}, booktitle={Pathways 2023: Human Dimensions of Wildlife Conference}, author={von Furstenberg, R. and Larson, L. and Rutledge, L.}, year={2023} }
 @inproceedings{evans_larson_millspaugh_brice_bashford_von furstenberg_2022, title={College R3 programs: Are they the answer to diversifying hunting participation?}, booktitle={The Wildlife Society Annual Conference}, author={Evans, C. and Larson, L. and Millspaugh, J. and Brice, J. and Bashford, B. and von Furstenberg, R.}, year={2022} }
 @inproceedings{larson_mengak_gramza_netherton-morrison_stinchcomb_von furstenberg_guilbeau_westlake_gray_2022, title={Conceptualizing and measuring relevancy: Using social science to broaden support for wildlife conservation}, booktitle={The Wildlife Society Annual Conference}, author={Larson, L. and Mengak, L. and Gramza, A. and Netherton-Morrison, H. and Stinchcomb, T. and von Furstenberg, R. and Guilbeau, K. and Westlake, S. and Gray, C.}, year={2022} }
 @inproceedings{von furstenberg_larson_vayer_peterson_lee_james_clark_2022, title={Developing and implementing R3 programs for college students: lessons learned from a nationwide project designed to diversify hunting}, booktitle={Southeastern Environment & Recreation Research Conference}, author={von Furstenberg, R. and Larson, L. and Vayer, V. and Peterson, N. and Lee, K.J. and James, W. and Clark, C.}, year={2022} }
 @inproceedings{lerose_casola_peterson_price_beall_von furstenberg_vaughn._pharr_barber_larson_2022, title={Evaluating Implicit Attitudes Towards Sharks}, booktitle={Pathways for Salmon: Human Dimensions of Wildlife Conference}, author={Lerose, C. and Casola, W. and Peterson, M.N. and Price, C.S. and Beall, J. and von Furstenberg, R. and Vaughn., A. and Pharr, L. and Barber, A. and Larson, L.}, year={2022} }
 @article{beall_pharr_von furstenberg_barber_casola_vaughn_peterson_larson_2022, title={The influence of YouTube videos on human tolerance of sharks}, volume={26}, ISSN={1367-9430 1469-1795}, url={http://dx.doi.org/10.1111/acv.12808}, DOI={10.1111/acv.12808}, abstractNote={AbstractSharks are often depicted in the media as violent killers that actively seek out opportunities to harm humans. This framing may impact human tolerance and support of shark conservation, underscoring the need to identify strategies that counteract these negative representations. Social media, given its widespread use, could be an effective platform for shaping public tolerance for sharks and other wildlife species. In this experimental study, we conducted an online pre‐post survey in Spring 2020 to determine how viewing shark‐related YouTube videos impacted tolerance for sharks among residents (n = 335) in the coastal state of North Carolina (NC), USA and neighboring states. The study employed framing theory, which suggests that the ways in which information is presented influence how it is processed and the actions that result from it. Participants were randomly assigned to one of two video treatments where sharks were framed positively or negatively. Each video treatment impacted tolerance for sharks in the direction of their framing: positive framing influenced positive changes in tolerance (70% more positive attitudes toward sharks, a 130% increase in acceptance of sharks and a 46% increase in intended shark conservation behaviors), and negative framing influenced negative changes (25% more negative attitudes toward sharks, a 18% decrease in acceptance of sharks and a 3% decrease in intended shark conservation behaviors). These findings suggest positive messages about sharks on social media promote tolerance of sharks and can be more impactful than negative messages. At least one form of social media, YouTube, appears to be a valuable tool for encouraging tolerance for sharks.}, number={2}, journal={Animal Conservation}, publisher={Wiley}, author={Beall, J. M. and Pharr, L. D. and von Furstenberg, R. and Barber, A. and Casola, W. R. and Vaughn, A. and Peterson, M. N. and Larson, L. R.}, year={2022}, month={Jul}, pages={154–164} }
 @inproceedings{von furstenberg_larson_peterson_2021, title={Developing and Implementing R3 Programs for College Students: Lessons Learned from a Nationwide Project Designed to Diversify Hunting}, booktitle={Meeting of the Southeastern Association of Fish and Wildlife Agencies}, author={von Furstenberg, R.J. and Larson, L.R. and Peterson, M.N.}, year={2021} }
 @article{vayer_larson_peterson_lee_von furstenberg_choi_stevenson_ahlers_anhalt‐depies_bethke_et al._2021, title={Diverse University Students Across the United States Reveal Promising Pathways to Hunter Recruitment and Retention}, volume={85}, ISSN={0022-541X 1937-2817}, url={http://dx.doi.org/10.1002/jwmg.22055}, DOI={10.1002/jwmg.22055}, abstractNote={ABSTRACTDeclining participation in hunting, especially among young adult hunters, affects the ability of state and federal agencies to achieve goals for wildlife management and decreases revenue for conservation. For wildlife agencies hoping to engage diverse audiences in hunter recruitment, retention, and reactivation (R3) efforts, university settings provide unique advantages: they contain millions of young adults who are developmentally primed to explore new activities, and they cultivate a social atmosphere where new identities can flourish. From 2018 to 2020, we surveyed 17,203 undergraduate students at public universities across 22 states in the United States to explore R3 potential on college campuses and assess key demographic, social, and cognitive correlates of past and intended future hunting behavior. After weighting to account for demographic differences between our sample and the larger student population, 29% of students across all states had hunted in the past. Students with previous hunting experience were likely to be white, male, from rural areas or hunting families, and pursuing degrees related to natural resources. When we grouped students into 1 of 4 categories with respect to hunting (i.e., non‐hunters [50%], potential hunters [22%], active hunters [26%], and lapsed hunters [3%]), comparisons revealed differences based on demographic attributes, beliefs, attitudes, and behaviors. Compared to active hunters, potential hunters were more likely to be females or racial and ethnic minorities, and less likely to experience social support for hunting. Potential hunters valued game meat and altruistic reasons for hunting, but they faced unique constraints due to lack of hunting knowledge and skills. Findings provide insights for marketing and programming designed to achieve R3 objectives with a focus on university students. © 2021 The Wildlife Society.}, number={5}, journal={The Journal of Wildlife Management}, publisher={Wiley}, author={Vayer, Victoria R. and Larson, Lincoln R. and Peterson, M. Nils and Lee, Kangjae Jerry and Von Furstenberg, Richard and Choi, Daniel Y. and Stevenson, Kathryn and Ahlers, Adam A. and Anhalt‐Depies, Christine and Bethke, Taniya and et al.}, year={2021}, month={May}, pages={1017–1030} }
 @inproceedings{von furstenberg_larson_peterson_2021, title={Social identities of U.S. college students reveal potential conflict and common ground for wildlife conservation}, booktitle={Annual Conference of The Wildlife Society}, author={von Furstenberg, R.J. and Larson, L.R. and Peterson, M.N.}, year={2021} }
 @article{larson_peterson_furstenberg_vayer_lee_choi_stevenson_ahlers_anhalt-depies_bethke_et al._2021, title={The future of wildlife conservation funding: What options do US college students support?}, volume={3}, ISSN={2578-4854 2578-4854}, url={http://dx.doi.org/10.1111/csp2.505}, DOI={10.1111/csp2.505}, abstractNote={AbstractInsufficient funding is a major impediment to conservation efforts around the world. In the United States, a decline in hunting participation threatens sustainability of the “user‐pay, public benefit” model that has supported wildlife conservation for nearly 100 years, forcing wildlife management agencies to contemplate alternative funding strategies. We investigated support for potential funding options among diverse college students, a rapidly expanding and politically active voting bloc with a potentially powerful influence on the future of conservation. From 2018 to 2020, we surveyed 17,203 undergraduate students at public universities across 22 states. Students preferred innovative approaches to conservation funding, with 72% supporting funding derived from industry sources (e.g., natural resource extraction companies), 63% supporting state sources (e.g., general sales tax), and 43% supporting conventional user‐based sources such as license fees and excise taxes associated with outdoor recreation activities (e.g., hunting). Findings emphasize the need to broaden the base of support for conservation funding and highlight the importance of considering the preferences and perspectives of young adults and other diverse beneficiaries of wildlife conservation.}, number={10}, journal={CONSERVATION SCIENCE AND PRACTICE}, publisher={Wiley}, author={Larson, Lincoln R. and Peterson, Markus Nils and Furstenberg, Richard Von and Vayer, Victoria R. and Lee, Kangjae Jerry and Choi, Daniel Y. and Stevenson, Kathryn and Ahlers, Adam A. and Anhalt-Depies, Christine and Bethke, Taniya and et al.}, year={2021}, month={Jul}, pages={e505} }
 @inproceedings{von furstenberg_larson_vayer_lee_peterson_2020, title={Rethinking R3: Broadening support for wildlife recreation with a focus on college students}, booktitle={Annual Conference of the Wildlife Society}, author={von Furstenberg, R.J. and Larson, L.R. and Vayer, V.R. and Lee, K.J. and Peterson, M.N.}, year={2020}, month={Oct} }
 @article{allena_von furstenberg_martinez-uribe_golubski_garman_2019, title={777 – Loss of TGF-Β Signaling Promotes Proliferation and is Associated with Barrett's Esophagus and Esophageal Adenocarcinoma}, volume={156}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(19)37193-8}, DOI={10.1016/s0016-5085(19)37193-8}, number={6}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Allena, Bhavya K. and von Furstenberg, Richard J. and Martinez-Uribe, Omar and Golubski, Bryan and Garman, Katherine S.}, year={2019}, month={May}, pages={S-159-S-160} }
 @article{boone_rochelle_ginzel_lubkov_roberts_nicholls_bock_flowers_von furstenberg_stripp_et al._2019, title={A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion}, volume={10}, ISSN={2041-1723}, url={http://dx.doi.org/10.1038/s41467-019-13330-y}, DOI={10.1038/s41467-019-13330-y}, abstractNote={Abstract Field cancerization is a premalignant process marked by clones of oncogenic mutations spreading through the epithelium. The timescales of intestinal field cancerization can be variable and the mechanisms driving the rapid spread of oncogenic clones are unknown. Here we use a Cancer rainbow (Crainbow) modelling system for fluorescently barcoding somatic mutations and directly visualizing the clonal expansion and spread of oncogenes. Crainbow shows that mutations of ß-catenin ( Ctnnb1 ) within the intestinal stem cell results in widespread expansion of oncogenes during perinatal development but not in adults. In contrast, mutations that extrinsically disrupt the stem cell microenvironment can spread in adult intestine without delay. We observe the rapid spread of premalignant clones in Crainbow mice expressing oncogenic Rspondin-3 ( RSPO3 ), which occurs by increasing crypt fission and inhibiting crypt fixation. Crainbow modelling provides insight into how somatic mutations rapidly spread and a plausible mechanism for predetermining the intratumor heterogeneity found in colon cancers.}, number={1}, journal={Nature Communications}, publisher={Springer Science and Business Media LLC}, author={Boone, Peter G. and Rochelle, Lauren K. and Ginzel, Joshua D. and Lubkov, Veronica and Roberts, Wendy L. and Nicholls, P. J. and Bock, Cheryl and Flowers, Mei Lang and von Furstenberg, Richard J. and Stripp, Barry R. and et al.}, year={2019}, month={Dec} }
 @article{hussain_von furstenberg_martinez-uribe_allena_becker_garman_2019, title={Su1040 – Early Activation of Sox9-Positive Cells in Esophageal Submucosal Glands After Injury}, volume={156}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(19)38094-1}, DOI={10.1016/s0016-5085(19)38094-1}, number={6}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Hussain, Farah S. and von Furstenberg, Richard J. and Martinez-Uribe, Omar and Allena, Bhavya K. and Becker, Thomas and Garman, Katherine S.}, year={2019}, month={May}, pages={S-492} }
 @article{martinez-uribe_von furstenberg_su_hussain_mccall_garman_2018, title={1075 - Increased Gastrin Responsiveness after Injury in Esophageal Submucosal Glands}, volume={154}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(18)31090-4}, DOI={10.1016/s0016-5085(18)31090-4}, number={6}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Martinez-Uribe, Omar and von Furstenberg, Richard J. and Su, Zuowei and Hussain, Farah S. and McCall, Shannon J. and Garman, Katherine S.}, year={2018}, month={May}, pages={S-207-S-208} }
 @article{iich_von furstenberg_garman_duong_clemons_phillip_2018, title={Su1762 - Esophageal Submucosal Glands as a Source of Progenitor Cells Capable of Differentiating Into Squamous or Barrett's-Like Columnar Cells}, volume={154}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(18)32125-5}, DOI={10.1016/s0016-5085(18)32125-5}, abstractNote={Background: Information for prediction factors of concurrent chemo-radiotherapy (CCRT) response has been limited.Primary 3-dimensional (3D) cultured cells have advantages in providing more physiologically relevant and predictive data for in-vivo response.Method: Primary 3D cell (spheroid) culture was performed using tumor tissues acquired from esophageal cancer before 1 st CCRT start.After 7days cultured, same sized spheroids were collected and were treated with 5-FU and 5Gy radiotherapy was provided.After 6days, primary cultured cells were stained and fluorescent images were captured.Clinical response was assessed after 4 th cycle CCRT.Clinical response was classified as complete remission (CR), partial remission (PR), and disease progression (PD).Results: A total of 26 esophageal cancer patients were enrolled.Final success rate of primary 3D cell culture was 77% (20/ 26).CCRT response in primary 3D cultured cell were evaluated in 20 cases.A total of 15 persons were followed up more than 4 cycles of CCRT and were analyzed.Clinical CR was observed in 9 persons and two persons showed clinical PR (n=4) or PD (n=2).Live activity was noted in less than 10% of primary 3D cultured cells in all patients with clinical CR and was observed in 30-40% of primary 3D cultured cells in all patients with clinical PD.Live activity was noted in less than 20-30% of primary 3D cultured cells in all patients with clinical PR Discussion: It takes 2weeks to evaluate the CCRT response in spheroid from tissue acquirement.High agreement between clinical response and response in spheroids was observed.The evaluation of CCRT response in spheroid will provide a good predictor of clinical CCRT response. Su1762}, number={6}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Iich, Elhadi and von Furstenberg, Richard J. and Garman, Katherine S. and Duong, Cuong and Clemons, Nicholas J. and Phillip, Wayne A.}, year={2018}, month={May}, pages={S-582} }
 @article{schoenborn_von furstenberg_valsaraj_hussain_stein_shanahan_henning_gulati_2018, title={The enteric microbiota regulates jejunal Paneth cell number and function without impacting intestinal stem cells}, volume={10}, ISSN={1949-0976 1949-0984}, url={http://dx.doi.org/10.1080/19490976.2018.1474321}, DOI={10.1080/19490976.2018.1474321}, abstractNote={Paneth cells (PCs) are epithelial cells found in the small intestine, next to intestinal stem cells (ISCs) at the base of the crypts. PCs secrete antimicrobial peptides (AMPs) that regulate the commensal gut microbiota. In contrast, little is known regarding how the enteric microbiota reciprocally influences PC function. In this study, we sought to characterize the impact of the enteric microbiota on PC biology in the mouse small intestine. This was done by first enumerating jejunal PCs in germ-free (GF) versus conventionally raised (CR) mice. We next evaluated the possible functional consequences of altered PC biology in these experimental groups by assessing epithelial proliferation, ISC numbers, and the production of AMPs. We found that PC numbers were significantly increased in CR versus GF mice; however, there were no differences in ISC numbers or cycling activity between groups. Of the AMPs assessed, only Reg3γ transcript expression was significantly increased in CR mice. Intriguingly, this increase was abrogated in cultured CR versus GF enteroids, and could not be re-induced with various bacterial ligands. Our findings demonstrate the enteric microbiota regulates PC function by increasing PC numbers and inducing Reg3γ expression, though the latter effect may not involve direct interactions between bacteria and the intestinal epithelium. In contrast, the enteric microbiota does not appear to regulate jejunal ISC census and proliferation. These are critical findings for investigators using GF mice and the enteroid system to study PC and ISC biology.}, number={1}, journal={Gut Microbes}, publisher={Informa UK Limited}, author={Schoenborn, Alexi A. and von Furstenberg, Richard J. and Valsaraj, Smrithi and Hussain, Farah S. and Stein, Molly and Shanahan, Michael T. and Henning, Susan J. and Gulati, Ajay S.}, year={2018}, month={Jul}, pages={45–58} }
 @article{krüger_gonzalez_pridgen_mccall_furstenberg_harnden_carnighan_cox_blikslager_garman_et al._2017, title={Ductular and proliferative response of esophageal submucosal glands in a porcine model of esophageal injury and repair}, volume={313}, ISSN={0193-1857 1522-1547}, url={http://dx.doi.org/10.1152/ajpgi.00036.2017}, DOI={10.1152/ajpgi.00036.2017}, abstractNote={ Esophageal injury is a risk factor for diseases such as Barrett’s esophagus (BE) and esophageal adenocarcinoma. To improve understanding of signaling pathways associated with both normal and abnormal repair, animal models are needed. Traditional rodent models of esophageal repair are limited by the absence of esophageal submucosal glands (ESMGs), which are present in the human esophagus. Previously, we identified acinar ductal metaplasia in human ESMGs in association with both esophageal injury and cancer. In addition, the SOX9 transcription factor has been associated with generation of columnar epithelium and the pathogenesis of BE and is present in ESMGs. To test our hypothesis that ESMGs activate after esophageal injury with an increase in proliferation, generation of a ductal phenotype, and expression of SOX9, we developed a porcine model of esophageal injury and repair using radiofrequency ablation (RFA). The porcine esophagus contains ESMGs, and RFA produces a consistent and reproducible mucosal injury in the esophagus. Here we present a temporal assessment of this model of esophageal repair. Porcine esophagus was evaluated at 0, 6, 18, 24, 48, and 72 h and 5 and 7 days following RFA and compared with control uninjured esophagus. Following RFA, ESMGs demonstrated an increase in ductal phenotype, echoing our prior studies in humans. Proliferation increased in both squamous epithelium and ESMGs postinjury with a prominent population of SOX9-positive cells in ESMGs postinjury. This model promises to be useful in future experiments evaluating mechanisms of esophageal repair.  NEW & NOTEWORTHY A novel porcine model of injury and repair using radiofrequency ablation has been developed, allowing for reproducible injury to the esophagus to study repair in an animal model with esophageal submucosal glands, a key anatomical feature and missing in rodent models but possibly harboring progenitor cells. There is a strong translational component to this porcine model given the anatomical and physiological similarities between pigs and humans. }, number={3}, journal={American Journal of Physiology-Gastrointestinal and Liver Physiology}, publisher={American Physiological Society}, author={Krüger, Leandi and Gonzalez, Liara and Pridgen, Tiffany A. and McCall, Shannon J. and Furstenberg, Richard J. and Harnden, Ivan and Carnighan, Gwendolyn E. and Cox, Abigail M. and Blikslager, Anthony and Garman, Katherine S. and et al.}, year={2017}, month={Sep}, pages={G180–G191} }
 @article{yan_gevaert_zheng_anchang_probert_larkin_davies_cheng_kaddis_han_et al._2017, title={Intestinal Enteroendocrine Lineage Cells Possess Homeostatic and Injury-Inducible Stem Cell Activity}, volume={21}, ISSN={1934-5909}, url={http://dx.doi.org/10.1016/j.stem.2017.06.014}, DOI={10.1016/j.stem.2017.06.014}, abstractNote={Several cell populations have been reported to possess intestinal stem cell (ISC) activity during homeostasis and injury-induced regeneration. Here, we explored inter-relationships between putative mouse ISC populations by comparative RNA-sequencing (RNA-seq). The transcriptomes of multiple cycling ISC populations closely resembled Lgr5+ ISCs, the most well-defined ISC pool, but Bmi1-GFP+ cells were distinct and enriched for enteroendocrine (EE) markers, including Prox1. Prox1-GFP+ cells exhibited sustained clonogenic growth in vitro, and lineage-tracing of Prox1+ cells revealed long-lived clones during homeostasis and after radiation-induced injury in vivo. Single-cell mRNA-seq revealed two subsets of Prox1-GFP+ cells, one of which resembled mature EE cells while the other displayed low-level EE gene expression but co-expressed tuft cell markers, Lgr5 and Ascl2, reminiscent of label-retaining secretory progenitors. Our data suggest that the EE lineage, including mature EE cells, comprises a reservoir of homeostatic and injury-inducible ISCs, extending our understanding of cellular plasticity and stemness.}, number={1}, journal={Cell Stem Cell}, publisher={Elsevier BV}, author={Yan, Kelley S. and Gevaert, Olivier and Zheng, Grace X.Y. and Anchang, Benedict and Probert, Christopher S. and Larkin, Kathryn A. and Davies, Paige S. and Cheng, Zhuan-fen and Kaddis, John S. and Han, Arnold and et al.}, year={2017}, month={Jul}, pages={78–90.e6} }
 @article{yan_janda_chang_zheng_larkin_luca_chia_mah_han_terry_et al._2017, title={Non-equivalence of Wnt and R-spondin ligands during Lgr5+ intestinal stem-cell self-renewal}, volume={545}, ISSN={0028-0836 1476-4687}, url={http://dx.doi.org/10.1038/nature22313}, DOI={10.1038/nature22313}, abstractNote={R-spondin and Wnt ligand families act non-redundantly and cooperatively within the same molecular pathway in the intestinal stem-cell niche to maintain stem-cell competency and drive stem-cell expansion. Wnt ligands interact with FZD and Lrp5/6-type receptors to influence diverse developmental, homeostatic and pathologic processes through β-catenin-dependent signalling. However, the promiscuity of Wnt ligands towards several receptors and the fact that Wnts can be hydrophobic make it difficult to produce therapeutic recombinant Wnts. Calvin Kuo and colleagues use a novel water-soluble Wnt agonist, developed by Chris Garcia and his team, in the mouse intestinal stem-cell niche to dissect the respective roles of R-spondin and Wnt ligands, both of which activate similar signalling receptors and pathways. They find that Lgr5+ intestinal stem cells normally differentiate unless both R-spondin and Wnt ligands are present. However, on their own, each ligand acts non-redundantly and in cooperation with Wnt agonist activating R-spondin receptors to maintain stem-cell competency.These receptorsare in turn activated in the presence of R-spondin to drive stem-cell expansion. Elsewhere in this issue, Chris Garcia and colleagues present their surrogate water-soluble Wnt agonists that have specificity towards certain FZDs.The new agonists act similarly to Wnt3 in differentiation assays towards the osteogenic lineage in vitro, can maintain intestinal organoid cultures, and have in vivo effects on the mouse liver. These water-soluble Wnt agonists could be used in a range of assays to understand this signalling pathway and modulate it in therapeutical applications. The canonical Wnt/β-catenin signalling pathway governs diverse developmental, homeostatic and pathological processes. Palmitoylated Wnt ligands engage cell-surface frizzled (FZD) receptors and LRP5 and LRP6 co-receptors, enabling β-catenin nuclear translocation and TCF/LEF-dependent gene transactivation1,2,3. Mutations in Wnt downstream signalling components have revealed diverse functions thought to be carried out by Wnt ligands themselves. However, redundancy between the 19 mammalian Wnt proteins and 10 FZD receptors1 and Wnt hydrophobicity have made it difficult to attribute these functions directly to Wnt ligands2,3. For example, individual mutations in Wnt ligands have not revealed homeostatic phenotypes in the intestinal epithelium4—an archetypal canonical, Wnt pathway-dependent, rapidly self-renewing tissue, the regeneration of which is fueled by proliferative crypt Lgr5+ intestinal stem cells (ISCs)5,6,7,8,9. R-spondin ligands (RSPO1–RSPO4) engage distinct LGR4–LGR6, RNF43 and ZNRF3 receptor classes10,11,12,13, markedly potentiate canonical Wnt/β-catenin signalling, and induce intestinal organoid growth in vitro and Lgr5+ ISCs in vivo8,14,15,16,17. However, the interchangeability, functional cooperation and relative contributions of Wnt versus RSPO ligands to in vivo canonical Wnt signalling and ISC biology remain unknown. Here we identify the functional roles of Wnt and RSPO ligands in the intestinal crypt stem-cell niche. We show that the default fate of Lgr5+ ISCs is to differentiate, unless both RSPO and Wnt ligands are present. However, gain-of-function studies using RSPO ligands and a new non-lipidated Wnt analogue reveal that these ligands have qualitatively distinct, non-interchangeable roles in ISCs. Wnt proteins are unable to induce Lgr5+ ISC self-renewal, but instead confer a basal competency by maintaining RSPO receptor expression that enables RSPO ligands to actively drive and specify the extent of stem-cell expansion. This functionally non-equivalent yet cooperative interaction between Wnt and RSPO ligands establishes a molecular precedent for regulation of mammalian stem cells by distinct priming and self-renewal factors, with broad implications for precise control of tissue regeneration.}, number={7653}, journal={Nature}, publisher={Springer Science and Business Media LLC}, author={Yan, Kelley S. and Janda, Claudia Y. and Chang, Junlei and Zheng, Grace X. Y. and Larkin, Kathryn A. and Luca, Vincent C. and Chia, Luis A. and Mah, Amanda T. and Han, Arnold and Terry, Jessica M. and et al.}, year={2017}, month={May}, pages={238–242} }
 @article{von furstenberg_li_stolarchuk_feder_campbell_kruger_gonzalez_blikslager_cardona_mccall_et al._2017, title={Porcine Esophageal Submucosal Gland Culture Model Shows Capacity for Proliferation and Differentiation}, volume={4}, ISSN={2352-345X}, url={http://dx.doi.org/10.1016/j.jcmgh.2017.07.005}, DOI={10.1016/j.jcmgh.2017.07.005}, abstractNote={<h3>Background & Aims</h3> Although cells comprising esophageal submucosal glands (ESMGs) represent a potential progenitor cell niche, new models are needed to understand their capacity to proliferate and differentiate. By histologic appearance, ESMGs have been associated with both overlying normal squamous epithelium and columnar epithelium. Our aim was to assess ESMG proliferation and differentiation in a 3-dimensional culture model. <h3>Methods</h3> We evaluated proliferation in human ESMGs from normal and diseased tissue by proliferating cell nuclear antigen immunohistochemistry. Next, we compared 5-ethynyl-2′-deoxyuridine labeling in porcine ESMGs in vivo before and after esophageal injury with a novel in vitro porcine organoid ESMG model. Microarray analysis of ESMGs in culture was compared with squamous epithelium and fresh ESMGs. <h3>Results</h3> Marked proliferation was observed in human ESMGs of diseased tissue. This activated ESMG state was recapitulated after esophageal injury in an in vivo porcine model, ESMGs assumed a ductal appearance with increased proliferation compared with control. Isolated and cultured porcine ESMGs produced buds with actively cycling cells and passaged to form epidermal growth factor–dependent spheroids. These spheroids were highly proliferative and were passaged multiple times. Two phenotypes of spheroids were identified: solid squamous (P63+) and hollow/ductal (cytokeratin 7+). Microarray analysis showed spheroids to be distinct from parent ESMGs and enriched for columnar transcripts. <h3>Conclusions</h3> Our results suggest that the activated ESMG state, seen in both human disease and our porcine model, may provide a source of cells to repopulate damaged epithelium in a normal manner (squamous) or abnormally (columnar epithelium). This culture model will allow the evaluation of factors that drive ESMGs in the regeneration of injured epithelium. The raw microarray data have been uploaded to the National Center for Biotechnology Information Gene Expression Omnibus (accession number: GSE100543).}, number={3}, journal={Cellular and Molecular Gastroenterology and Hepatology}, publisher={Elsevier BV}, author={von Furstenberg, Richard J. and Li, Joy and Stolarchuk, Christina and Feder, Rachel and Campbell, Alexa and Kruger, Leandi and Gonzalez, Liara M. and Blikslager, Anthony T. and Cardona, Diana M. and McCall, Shannon J. and et al.}, year={2017}, month={Nov}, pages={385–404} }
 @article{schoenborn_von furstenberg_valsaraj_hussain_stein_shanahan_gulati_henning_2017, title={The Enteric Microbiota Regulates Paneth Cell Number and Function Without Affecting Intestinal Stem Cells}, volume={152}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(17)30418-3}, DOI={10.1016/s0016-5085(17)30418-3}, number={5}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Schoenborn, Alexi A. and von Furstenberg, Richard and Valsaraj, Smrithi and Hussain, Farah S. and Stein, Molly and Shanahan, Michael and Gulati, Ajay S. and Henning, Susan J.}, year={2017}, month={Apr}, pages={S13} }
 @article{henning_von furstenberg_2016, title={GI stem cells – new insights into roles in physiology and pathophysiology}, volume={594}, ISSN={0022-3751 1469-7793}, url={http://dx.doi.org/10.1113/jp271663}, DOI={10.1113/jp271663}, abstractNote={This overview gives a brief historical summary of key discoveries regarding stem cells of the small intestine. The current concept is that there are two pools of intestinal stem cells (ISCs): an actively cycling pool that is marked by Lgr5, is relatively homogeneous and is responsible for daily turnover of the epithelium; and a slowly cycling or quiescent pool that functions as reserve ISCs. The latter pool appears to be quite heterogeneous and may include partially differentiated epithelial lineages that can reacquire stem cell characteristics following injury to the intestine. Markers and methods of isolation for active and quiescent ISC populations are described as well as the numerous important advances that have been made in approaches to the in vitro culture of ISCs and crypts. Factors regulating ISC biology are briefly summarized and both known and unknown aspects of the ISC niche are discussed. Although most of our current knowledge regarding ISC physiology and pathophysiology has come from studies with mice, recent work with human tissue highlights the potential translational applications arising from this field of research. Many of these topics are further elaborated in the following articles.}, number={17}, journal={The Journal of Physiology}, publisher={Wiley}, author={Henning, Susan J. and von Furstenberg, Richard J.}, year={2016}, month={Apr}, pages={4769–4779} }
 @article{kruger_gonzalez_von furstenberg_henning_blikslager_garman_2016, title={Mo1253 Ductular and Proliferative Response of Esophageal Submucosal Glands in a Porcine Model of Esophageal Injury and Repair}, volume={150}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(16)32312-5}, DOI={10.1016/s0016-5085(16)32312-5}, number={4}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Kruger, Leandi and Gonzalez, Liara M. and von Furstenberg, Richard J. and Henning, Susan J. and Blikslager, Anthony and Garman, Katherine S.}, year={2016}, month={Apr}, pages={S679–S680} }
 @article{garman_boutte_von furstenberg_chiu_lloyd_zhang_onaitis_chow_mccall_2016, title={Tu1179 Previous Tonsillectomy Is Associated With Increased Risk of Esophageal Cancer}, volume={150}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(16)32889-x}, DOI={10.1016/s0016-5085(16)32889-x}, number={4}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Garman, Katherine S. and Boutte, Harold J. and von Furstenberg, Richard J. and Chiu, Shih-Ting and Lloyd, Benjamin and Zhang, Cecelia and Onaitis, Mark and Chow, Shein-Chung and McCall, Shannon J.}, year={2016}, month={Apr}, pages={S856} }
 @inproceedings{von furstenberg_li_stolarchuk_feder_campbell_kruger_gonzalez_blikslager_mccall_henning_et al._2015, title={Esophageal submucosal gland culture model demonstrates capacity for proliferation and differentiation}, booktitle={CMGH James W Freston Single Topic Conference}, author={von Furstenberg, Richard J. and Li, Joy and Stolarchuk, Christina and Feder, Rachel and Campbell, Alexa and Kruger, Leandi and Gonzalez, Liara M. and Blikslager, Anthony T. and McCall, Shannon J. and Henning, Susan J. and et al.}, year={2015} }
 @article{davies_santaolalla_von furstenberg_henning_abreu_2015, title={Mo1836 Viral Products Elicit Different Phenotypes From Small Intestine and Colonic Crypt Cultures}, volume={148}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(15)32465-3}, DOI={10.1016/s0016-5085(15)32465-3}, abstractNote={The small intestine and colon differ greatly in both function and exposure to microbial products.Specific studies are needed to assess the response of microbial signaling in the small intestine versus the colon.Double-stranded RNAs (ds-RNA) are primarily viral products, but can also be derived from bacteria.Acute enteric viruses usually impact the small intestine, but viruses have also been investigated in inflammatory bowel disease and in post-infective irritable bowel syndrome and viruses are frequently found in colon cancer tissue.As such, understanding differences in the response of epithelial cells from the small intestine and colon to exposure to viral products will be important for dissecting the impact of infection at the different anatomical locations.Using newly developed culture techniques for growing primary enteroids and colonoids, we sought to compare the responses of enteroids versus colonoids to stimulation with the synthetic ds-RNA polyinosinic-polycytidylic acid (poly I:C).Murine crypts from jejunum or colon were plated in Matrigel for two days prior to the addition of poly I:C for the duration of culture.Stimulation of enteroids with poly I:C significantly altered the surface area but decreased the number of surviving enteroids compared to unstimulated.In colonoids, stimulation with poly I:C resulted in a significant decrease in bud count, but did not impact the area or survival of the organoids.Gene expression measured by the probe-based Nanostring technology in enteroids following poly I:C stimulation showed significantly decreased expression of stem cell markers including: Sox9, Lgr5, Dclk1, Tert and Lrig1.Additionally, differentiation markers Sis and Muc2 were significantly decreased.In contrast, fewer genes were significantly impacted by poly I:C stimulation in colonoids.Stem cell markers were not altered by poly I:C stimulation in colonoids.As expected, in both enteroids and colonoids inflammatory markers were significantly increased in response to poly I:C stimulation.Additionally, pro-apoptotic genes in both enteroids and colonoids were decreased following poly I:C stimulation.Comparing the response of enteroids and colonoids to poly I:C stimulation we found differences in the magnitude or direction of change in several classes of genes.These distinctions may derive from baseline differences in expression levels.Enteroids had significantly increased expression of stem cell markers at baseline compared to colonoids, while colonoids had increased levels of inflammatory markers and toll-like receptors.Together, these data indicate that important functional differences exist between enteroids and colonoids at baseline and after viral product stimulation providing evidence which may help explain the different anatomical responses to viral infection throughout the intestinal tract.}, number={4}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Davies, Julie M. and Santaolalla, Rebeca and von Furstenberg, Richard J. and Henning, Susan J. and Abreu, Maria T.}, year={2015}, month={Apr}, pages={S-723} }
 @article{davies_santaolalla_von furstenberg_henning_abreu_2015, title={The Viral Mimetic Polyinosinic:Polycytidylic Acid Alters the Growth Characteristics of Small Intestinal and Colonic Crypt Cultures}, volume={10}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0138531}, DOI={10.1371/journal.pone.0138531}, abstractNote={The intestinal epithelium is the first line of defense against enteric pathogens. We investigated the response of small intestinal and colonic crypt cultures to a panel of toll-like receptor ligands to assess the impact of microbial pattern recognition on epithelial growth.Primary murine jejunal enteroids and colonoids were cultured with lipopeptide Pam3CSK4, lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (Poly I:C) for 4 to 6 days. Surface area, budding and survival were assessed. Proliferation and numbers of lysozyme positive cells were quantified by flow cytometry. Gene expression was assessed by Nanostring and qRT-PCR.Exposure to Pam3CSK4 and LPS had minimal impact on either enteroids or colonoids. In contrast, Poly I:C increased the surface area of enteroids, while colonoids demonstrated decreased budding. Survival was decreased by Poly I:C in enteroids but not in colonoids. Both enteroids and colonoids exhibited upregulated gene expression of chemokines, but these were increased in magnitude in enteroids. Decreases in gene expression associated with epithelial differentiation and lysozyme positive cells were more apparent in enteroids than in colonoids. Baseline gene expression between enteroids and colonoids differed markedly in levels of stem cell and inflammatory markers. The changes in morphology induced by Poly I:C were mediated by the toll-like receptor adaptor molecule 1 (Ticam1) in enteroids but not in colonoids.Poly I:C alters the molecular program of epithelial cells and shifts from absorption and digestion towards defense and inflammation. Diversity of responses to microbial patterns in enteroids and colonoids may underlie differences in susceptibility to infection along the intestinal tract.}, number={9}, journal={PLOS ONE}, publisher={Public Library of Science (PLoS)}, author={Davies, Julie M. and Santaolalla, Rebeca and von Furstenberg, Richard J. and Henning, Susan J. and Abreu, Maria T.}, editor={Boone, David LEditor}, year={2015}, month={Sep}, pages={e0138531} }
 @article{seiler_schenhals_von furstenberg_allena_smith_scaria_bresler_dekaney_henning_2015, title={Tissue underlying the intestinal epithelium elicits proliferation of intestinal stem cells following cytotoxic damage}, volume={361}, ISSN={0302-766X 1432-0878}, url={http://dx.doi.org/10.1007/s00441-015-2111-1}, DOI={10.1007/s00441-015-2111-1}, abstractNote={The goals of this study were to document the proliferative response of intestinal stem cells (ISCs) during regeneration after damage from doxorubicin (DXR), and to characterize the signals responsible for ISC activation. To this end, jejuni from DXR-treated mice were harvested for histology, assessment of ISC numbers and proliferation by flow cytometry, crypt culture, and RNA analyses. Histology showed that crypt depth and width were increased 4 days after DXR. At this time point, flow cytometry on tissue collected 1 h after EdU administration revealed increased numbers of CD24loUEA− ISCs and increased percentage of ISCs cycling. In culture, crypts harvested from DXR-treated mice were equally proliferative as those of control mice. Addition of subepithelial intestinal tissue (SET) collected 4 days after DXR elicited increased budding (1.4 ± 0.3 vs. 5.1 ± 1.0 buds per enteroid). Microarray analysis of SET collected 4 days after DXR revealed 1030 differentially expressed transcripts. Cross-comparison of Gene Ontology terms considered relevant to ISC activation pointed to 10 candidate genes. Of these, the epidermal growth factor (EGF) family member amphiregulin and the BMP antagonist chordin-like 2 were chosen for further study. In crypt culture, amphiregulin alone did not elicit significant budding, but amphiregulin in combination with BMP antagonism showed marked synergism (yielding 6.3 ± 0.5 buds per enteroid). These data suggest a critical role for underlying tissue in regulating ISC behavior after damage, and point to synergism between amphiregulin and chordin-like 2 as factors which may account for activation of ISCs in the regenerative phase.}, number={2}, journal={Cell and Tissue Research}, publisher={Springer Science and Business Media LLC}, author={Seiler, Kristen M and Schenhals, Erica L and von Furstenberg, Richard J and Allena, Bhavya K and Smith, Brian J and Scaria, Denny and Bresler, Michele N and Dekaney, Christopher M and Henning, Susan J}, year={2015}, month={Feb}, pages={427–438} }
 @article{von furstenberg_buczacki_smith_seiler_winton_henning_2014, title={Side population sorting separates subfractions of cycling and non-cycling intestinal stem cells}, volume={12}, ISSN={1873-5061}, url={http://dx.doi.org/10.1016/j.scr.2013.10.012}, DOI={10.1016/j.scr.2013.10.012}, abstractNote={We report here that side population (SP) sorting allows for the simultaneous isolation of two intestinal stem cell (ISC) subsets from wild-type (WT) mice which are phenotypically different and represent cycling and non-cycling pools of cells. Following 5-ethynyl-2′-deoxyuridine (EdU) injection, in the upper side population (USP) the percentage of EdU + was 36% showing this fraction to be highly proliferative. In the lower side population (LSP), only 0.4% of cells were EdU +, indicating this fraction to be predominantly non-cycling. Using Lgr5-EGFP mice, we show that Lgr5-EGFPhi cells, representing actively cycling ISCs, are essentially exclusive to the USP. In contrast, using histone 2B-YFP mice, SP analysis revealed YFP label retaining cells (LRCs) in both the USP and the LSP. Correspondingly, evaluation of the SP fractions for mRNA markers by qRT-PCR showed that the USP was enriched in transcripts associated with both quiescent and active ISCs. In contrast, the LSP expressed mRNA markers of quiescent ISCs while being de-enriched for those of the active ISC. Both the USP and LSP are capable of generating enteroids in culture which include the four intestinal lineages. We conclude that sorting of USP and LSP fractions represents a novel isolation of cycling and non-cycling ISCs from WT mice.}, number={2}, journal={Stem Cell Research}, publisher={Elsevier BV}, author={von Furstenberg, Richard J. and Buczacki, Simon J.A. and Smith, Brian J. and Seiler, Kristen M. and Winton, Douglas J. and Henning, Susan J.}, year={2014}, month={Mar}, pages={364–375} }
 @article{von furstenberg_buczacki_smith_winton_henning_2013, title={277 Side Population Analysis of Mouse Jejunal Epithelium Reveals Sub-Groups With Active and Quiescent Intestinal Stem Cell Phenotypes}, volume={144}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(13)60223-1}, DOI={10.1016/s0016-5085(13)60223-1}, number={5}, journal={Gastroenterology}, publisher={Elsevier BV}, author={von Furstenberg, Richard J. and Buczacki, Simon J. and Smith, Brian J. and Winton, Doug J. and Henning, Susan J.}, year={2013}, month={May}, pages={S-62} }
 @article{fuller_faulk_sundaram_mahe_stout_von furstenberg_smith_mcnaughton_shroyer_helmrath_et al._2013, title={Intestinal stem cells remain viable after prolonged tissue storage}, volume={354}, ISSN={0302-766X 1432-0878}, url={http://dx.doi.org/10.1007/s00441-013-1674-y}, DOI={10.1007/s00441-013-1674-y}, abstractNote={Intestinal stem cells (ISCs) are responsible for renewal of the epithelium both during normal homeostasis and following injury. As such, they have significant therapeutic potential. However, whether ISCs can survive tissue storage is unknown. We hypothesize that, although the majority of epithelial cells might die, ISCs would remain viable for at least 24 h at 4 °C. To explore this hypothesis, jejuna of C57Bl6/J or Lgr5-LacZ mice were removed and either processed immediately or placed in phosphate-buffered saline at 4 °C. Delayed isolation of epithelium was performed after 24, 30, or 48 h storage. At the light microscope level, despite extensive apoptosis of villus epithelial cells, small intestinal crypts remained morphologically intact for 30 h and ISCs were identifiable via Lgr5-LacZ positivity. Electron microscopy showed that ISCs retained high integrity for 24 h. When assessed by flow cytometry, ISCs were more resistant to degeneration than the rest of the epithelium, including neighboring Paneth cells, with higher viability across all time points. Cultured isolated crypts showed no loss of capacity to form complex enteroids after 24 h tissue storage, with efficiencies after 7 days of culture remaining above 80 %. By 30 h storage, efficiencies declined but budding capability was retained. We conclude that, with delay in isolation, ISCs remain viable and retain their proliferative capacity. In contrast, the remainder of the epithelium, including the Paneth cells, exhibits degeneration and programmed cell death. If these findings are recapitulated in human tissue, storage at 4 °C might offer a valuable temporal window for the harvesting of crypts or ISCs for therapeutic application.}, number={2}, journal={Cell and Tissue Research}, publisher={Springer Science and Business Media LLC}, author={Fuller, Megan K. and Faulk, Denver M. and Sundaram, Nambirajan and Mahe, Maxime M. and Stout, Kara M. and von Furstenberg, Richard J. and Smith, Brian J. and McNaughton, Kirk K. and Shroyer, Noah F. and Helmrath, Michael A. and et al.}, year={2013}, month={Jul}, pages={441–450} }
 @article{shanahan_carroll_grossniklaus_white_von furstenberg_barner_fodor_henning_sartor_gulati_2013, title={Mouse Paneth cell antimicrobial function is independent of Nod2}, volume={63}, ISSN={0017-5749 1468-3288}, url={http://dx.doi.org/10.1136/gutjnl-2012-304190}, DOI={10.1136/gutjnl-2012-304190}, abstractNote={Although polymorphisms of the NOD2 gene predispose to the development of ileal Crohn's disease, the precise mechanisms of this increased susceptibility remain unclear. Previous work has shown that transcript expression of the Paneth cell (PC) antimicrobial peptides (AMPs) α-defensin 4 and α-defensin-related sequence 10 are selectively decreased in Nod2(-/-) mice. However, the specific mouse background used in this previous study is unclear. In light of recent evidence suggesting that mouse strain strongly influences PC antimicrobial activity, we sought to characterise PC AMP function in commercially available Nod2(-/-) mice on a C57BL/6 (B6) background. Specifically, we hypothesised that Nod2(-/-) B6 mice would display reduced AMP expression and activity.Wild-type (WT) and Nod2(-/-) B6 ileal AMP expression was assessed via real-time PCR, acid urea polyacrylamide gel electrophoresis and mass spectrometry. PCs were enumerated using flow cytometry. Functionally, α-defensin bactericidal activity was evaluated using a gel-overlay antimicrobial assay. Faecal microbial composition was determined using 454-sequencing of the bacterial 16S gene in cohoused WT and Nod2(-/-) littermates.WT and Nod2(-/-) B6 mice displayed similar PC AMP expression patterns, equivalent α-defensin profiles, and identical antimicrobial activity against commensal and pathogenic bacterial strains. Furthermore, minimal differences in gut microbial composition were detected between the two cohoused, littermate mouse groups.Our data reveal that Nod2 does not directly regulate PC antimicrobial activity in B6 mice. Moreover, we demonstrate that previously reported Nod2-dependent influences on gut microbial composition may be overcome by environmental factors, such as cohousing with WT littermates.}, number={6}, journal={Gut}, publisher={BMJ}, author={Shanahan, Michael T and Carroll, Ian M and Grossniklaus, Emily and White, Andrew and von Furstenberg, Richard J and Barner, Roshonda and Fodor, Anthony A and Henning, Susan J and Sartor, R Balfour and Gulati, Ajay S}, year={2013}, month={Mar}, pages={903–910} }
 @article{king_von furstenberg_smith_mcnaughton_galanko_henning_2012, title={CD24 can be used to isolate Lgr5<sup>+</sup> putative colonic epithelial stem cells in mice}, volume={303}, ISSN={0193-1857 1522-1547}, url={http://dx.doi.org/10.1152/ajpgi.00087.2012}, DOI={10.1152/ajpgi.00087.2012}, abstractNote={A growing body of evidence has implicated CD24, a cell-surface protein, as a marker of colorectal cancer stem cells and target for antitumor therapy, although its presence in normal colonic epithelium has not been fully characterized. Previously, our group showed that CD24-based cell sorting can be used to isolate a fraction of murine small intestinal epithelial cells enriched in actively cycling stem cells. Similarly, we hypothesized that CD24-based isolation of colonic epithelial cells would generate a fraction enriched in actively cycling colonic epithelial stem cells (CESCs). Immunohistochemistry performed on mouse colonic tissue showed CD24 expression in the bottom half of proximal colon crypts and the crypt base in the distal colon. This pattern of distribution was similar to enhanced green fluorescent protein (EGFP) expression in Lgr5-EGFP mice. Areas expressing CD24 contained actively proliferating cells as determined by ethynyl deoxyuridine (EdU) incorporation, with a distinct difference between the proximal colon, where EdU-labeled cells were most frequent in the midcrypt, and the distal colon, where they were primarily at the crypt base. Flow cytometric analyses of single epithelial cells, identified by epithelial cell adhesion molecule (EpCAM) positivity, from mouse colon revealed an actively cycling CD24(+) fraction that contained the majority of Lgr5-EGFP(+) putative CESCs. Transcript analysis by quantitative RT-PCR confirmed enrichment of active CESC markers [leucine-rich-repeat-containing G protein-coupled receptor 5 (Lgr5), ephrin type B receptor 2 (EphB2), and CD166] in the CD24(+)EpCAM(+) fraction but also showed enrichment of quiescent CESC markers [leucine-rich repeats and immunoglobin domains (Lrig), doublecortin and calmodulin kinase-like 1 (DCAMKL-1), and murine telomerase reverse transcriptase (mTert)]. We conclude that CD24-based sorting in wild-type mice isolates a colonic epithelial fraction highly enriched in actively cycling and quiescent putative CESCs. Furthermore, the presence of CD24 expression in normal colonic epithelium may have important implications for the use of anti-CD24-based colorectal cancer therapies.}, number={4}, journal={American Journal of Physiology-Gastrointestinal and Liver Physiology}, publisher={American Physiological Society}, author={King, Jeffrey B. and von Furstenberg, Richard J. and Smith, Brian J. and McNaughton, Kirk K. and Galanko, Joseph A. and Henning, Susan J.}, year={2012}, month={Aug}, pages={G443–G452} }
 @inproceedings{fuller_faulk_stout_smith_von furstenberg_henning_helmrath_2012, title={Intestinal Stem Cells Are Resistant to Degeneration}, booktitle={2012 AAP National Conference and Exhibition}, author={Fuller, M.K. and Faulk, D.M. and Stout, K.M. and Smith, B.J. and von Furstenberg, R.J. and Henning, S.J. and Helmrath, M.A.}, year={2012}, month={Oct} }
 @article{gulati_shanahan_arthur_grossniklaus_von furstenberg_kreuk_henning_jobin_sartor_2012, title={Mouse Background Strain Profoundly Influences Paneth Cell Function and Intestinal Microbial Composition}, volume={7}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0032403}, DOI={10.1371/journal.pone.0032403}, abstractNote={Background Increasing evidence supports the central role of Paneth cells in maintaining intestinal host-microbial homeostasis. However, the direct impact of host genotype on Paneth cell function remains unclear. Here, we characterize key differences in Paneth cell function and intestinal microbial composition in two widely utilized, genetically distinct mouse strains (C57BL/6 and 129/SvEv). In doing so, we demonstrate critical influences of host genotype on Paneth cell activity and the enteric microbiota. Methodology and Principal Findings Paneth cell numbers were determined by flow cytometry. Antimicrobial peptide (AMP) expression was evaluated using quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR), acid urea-polyacrylamide gel electrophoresis, and mass spectrometry. Effects of mouse background on microbial composition were assessed by reciprocal colonization of germ-free mice from both background strains, followed by compositional analysis of resultant gut bacterial communities using terminal restriction fragment length polymorphism analysis and 16 S qPCR. Our results revealed that 129/SvEv mice possessed fewer Paneth cells and a divergent AMP profile relative to C57BL/6 counterparts. Novel 129/SvEv á-defensin peptides were identified, including Defa2/18v, Defa11, Defa16, and Defa18. Host genotype profoundly affected the global profile of the intestinal microbiota, while both source and host factors were found to influence specific bacterial groups. Interestingly, ileal α-defensins from 129/SvEv mice displayed attenuated antimicrobial activity against pro-inflammatory E. coli strains, a bacterial species found to be expanded in these animals. Conclusions and Significance This work establishes the important impact of host genotype on Paneth cell function and the composition of the intestinal microbiota. It further identifies specific AMP and microbial alterations in two commonly used inbred mouse strains that have varying susceptibilities to a variety of disorders, ranging from obesity to intestinal inflammation. This will be critical for future studies utilizing these murine backgrounds to study the effects of Paneth cells and the intestinal microbiota on host health and disease.}, number={2}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Gulati, Ajay S. and Shanahan, Michael T. and Arthur, Janelle C. and Grossniklaus, Emily and von Furstenberg, Richard J. and Kreuk, Lieselotte and Henning, Susan J. and Jobin, Christian and Sartor, R. Balfour}, editor={Bereswill, StefanEditor}, year={2012}, month={Feb}, pages={e32403} }
 @article{shanahan_grossniklaus_von furstenberg_henning_sartor_gulati_2012, title={Tu1856 NOD2 Does Not Regulate Mouse Paneth Cell a-Defensin Expression}, volume={142}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(12)63343-5}, DOI={10.1016/s0016-5085(12)63343-5}, abstractNote={infection, viability of TF, binding competition, cytoskeletal inhibitors (cytochalasin B, 10.5μM; colchicine, 250μM), sialic acid and protease inhibitors (E64, 300-600μM; EDTA, 500μM) were quantified by liquid scintillation counting and immunofluorescence of adherent Tf.A minimum of four replicates were performed for each experiment.Data were analyzed using commercially available statistical software.3[H]thymidine and CFSE-labeled Tf cytoadhered to IPEC-J2 monolayers in a dose and timedependent manner with reproducible adhesion achieved by infection with 20x106 Tf for 6hrs.Clinical isolates of feline Tf (n=5) demonstrated significantly greater adhesion (447,109 ± 48,248 trophozoites) to intestinal epithelium than Pentatrichomonas hominis (40,426 ± 9,877 trophozoites), the latter of which is a presumably nonpathogenic trichomonad of domestic animals and people.Adhesion of Tf required viable trophozoites but was independent of cytoskeletal activity.Based on competition binding experiments, adhesion of Tf was receptor-ligand dependent.Treatment of Tf with N-acetylneuraminic acid and cysteine protease inhibitors significantly blocked adherence.Identification of specific ligand-receptor dependent mechanisms of Tf adherence that are ameliorated by exogenous sialic acid and cysteine protease inhibitors suggests the presence of key molecular targets for prevention of Tf adhesion and consequent cytopathogenicity.The results of this work have comparative implications for future pharmacologic intervention against enteric protozoal pathogens and such investigations can be undertaken using a feline model of clinical infection.}, number={5}, journal={Gastroenterology}, publisher={Elsevier BV}, author={Shanahan, Michael T. and Grossniklaus, Emily and von Furstenberg, Richard J. and Henning, Susan J. and Sartor, Ryan B. and Gulati, Ajay S.}, year={2012}, month={May}, pages={S-862} }
 @article{von furstenberg_gulati_stappenbeck_henning_2010, title={787 Sorting With CD24 Yields a Population Markedly Enriched for Markers of Intestinal Stem Cells}, volume={138}, ISSN={0016-5085}, url={http://dx.doi.org/10.1016/s0016-5085(10)60508-2}, DOI={10.1016/s0016-5085(10)60508-2}, number={5}, journal={Gastroenterology}, publisher={Elsevier BV}, author={von Furstenberg, Richard J. and Gulati, Ajay S. and Stappenbeck, Thaddeus S. and Henning, Susan J.}, year={2010}, month={May}, pages={S-111} }
 @article{von furstenberg_gulati_baxi_doherty_stappenbeck_gracz_magness_henning_2011, title={Sorting mouse jejunal epithelial cells with CD24 yields a population with characteristics of intestinal stem cells}, volume={300}, ISSN={0193-1857 1522-1547}, url={http://dx.doi.org/10.1152/ajpgi.00453.2010}, DOI={10.1152/ajpgi.00453.2010}, abstractNote={Intestinal stem cells (ISCs) have been studied for more than three decades; however, their isolation has remained a challenge. We hypothesized that, just as for stem cells of other tissues, one or more membrane markers would allow positive selection of ISCs by antibody-based sorting. To explore this hypothesis, microarray data of putative ISC fractions generated by side population sorting and laser capture microdissection were subjected to bioinformatic analysis to identify common membrane antigens. The microarray comparison suggested CD24 as a candidate surface marker, and immunohistochemistry showed expression of CD24 in epithelial cells of crypt bases. Flow cytometry of jejunal epithelial preparations revealed a CD24 + CD45 − fraction comprising ∼1% of the cells. Analysis with epithelial cell adhesion molecule and CD31 confirmed that the cell preparations were epithelial and without endothelial contamination. Cycling cells identified by prior injection with 5-ethynyl-2′-deoxyuridine were found predominantly in the CD24 lo subfraction. Transcript analysis by real-time RT-PCR showed this subfraction to be enriched in the ISC markers leucine-rich-repeat-containing G-protein-coupled receptor 5 (40-fold) and Bmi1 (5-fold), but also enriched in lysozyme (10-fold). Flow cytometry with anti-lysozyme antibodies demonstrated that Paneth cells comprise ∼30% of the CD24 lo subfraction. Additional flow analyses with leucine-rich-repeat-containing G-protein-coupled receptor 5-enhanced green fluorescent protein (EGFP) epithelium demonstrated colocalization of EGFP hi and CD24 lo . In contrast, CD24 cells were negative for the quiescent ISC marker doublecortin and CaM kinase-like-1. Culture of CD24 lo cells in Matrigel generated organoid structures, which included all four epithelial lineages, thus giving functional evidence for the presence of ISCs. We conclude that the CD24 lo fraction of jejunal epithelium is highly enriched with cycling ISCs. This isolation method should be useful to many investigators in the field to advance both the basic understanding of ISC biology and the therapeutic applications of ISCs.}, number={3}, journal={American Journal of Physiology-Gastrointestinal and Liver Physiology}, publisher={American Physiological Society}, author={von Furstenberg, Richard J. and Gulati, Ajay S. and Baxi, Anand and Doherty, Jason M. and Stappenbeck, Thaddeus S. and Gracz, Adam D. and Magness, Scott T. and Henning, Susan J.}, year={2011}, month={Mar}, pages={G409–G417} }
 @article{ghose_mulder_von furstenberg_thevananther_kuipers_karpen_2007, title={Rosiglitazone attenuates suppression of RXRα-dependent gene expression in inflamed liver}, volume={46}, ISSN={0168-8278}, url={http://dx.doi.org/10.1016/j.jhep.2006.09.008}, DOI={10.1016/j.jhep.2006.09.008}, abstractNote={Background/Aims A recently determined target of lipopolysaccharide (LPS) and cytokine signaling in liver is the central Type II nuclear receptor (NR) heterodimer partner, retinoid X receptor α (RXRα). We sought to determine if Rosiglitazone (Rosi), a peroxisome proliferator activated receptor γ (PPARγ) agonist with anti-inflammatory properties, can attenuate LPS and cytokine-induced molecular suppression of RXRα-regulated genes. Methods In vivo, mice were gavage-fed Rosi for 3 days, prior to intraperitoneal injection of LPS, followed by harvest of liver and serum. In vitro, HepG2 cells were treated with IL-1β, ± short-term Rosi pretreatment. RNA was analyzed by quantitative RT-PCR, while nuclear and cytoplasmic proteins were analyzed by immunoblotting and gel shifts. Results Rosi attenuated LPS-mediated suppression of RNA levels of several Type II NR-regulated genes, including bile acid transporters and the major drug metabolizing enzyme, Cyp3a11, without affecting cytokine expression, suggesting a novel, direct anti-inflammatory effect in hepatocytes. Rosi suppressed the inflammation-induced nuclear export of RXRα, in both LPS-injected mice and IL-1β-treated HepG2 cells, leading to maintenance of nuclear RXRα levels and heterodimer binding activity. Conclusions Rosi directly attenuates the suppressive effects of inflammation-induced cell signaling on nuclear RXRα levels in liver. A recently determined target of lipopolysaccharide (LPS) and cytokine signaling in liver is the central Type II nuclear receptor (NR) heterodimer partner, retinoid X receptor α (RXRα). We sought to determine if Rosiglitazone (Rosi), a peroxisome proliferator activated receptor γ (PPARγ) agonist with anti-inflammatory properties, can attenuate LPS and cytokine-induced molecular suppression of RXRα-regulated genes. In vivo, mice were gavage-fed Rosi for 3 days, prior to intraperitoneal injection of LPS, followed by harvest of liver and serum. In vitro, HepG2 cells were treated with IL-1β, ± short-term Rosi pretreatment. RNA was analyzed by quantitative RT-PCR, while nuclear and cytoplasmic proteins were analyzed by immunoblotting and gel shifts. Rosi attenuated LPS-mediated suppression of RNA levels of several Type II NR-regulated genes, including bile acid transporters and the major drug metabolizing enzyme, Cyp3a11, without affecting cytokine expression, suggesting a novel, direct anti-inflammatory effect in hepatocytes. Rosi suppressed the inflammation-induced nuclear export of RXRα, in both LPS-injected mice and IL-1β-treated HepG2 cells, leading to maintenance of nuclear RXRα levels and heterodimer binding activity. Rosi directly attenuates the suppressive effects of inflammation-induced cell signaling on nuclear RXRα levels in liver.}, number={1}, journal={Journal of Hepatology}, publisher={Elsevier BV}, author={Ghose, Romi and Mulder, Jaap and von Furstenberg, Richard J. and Thevananther, Sundararajah and Kuipers, Folkert and Karpen, Saul J.}, year={2007}, month={Jan}, pages={115–123} }
 @article{carter_taylor_prendergast_zimmerman_von furstenberg_moore_karpen_2007, title={Stigmasterol, a Soy Lipid–Derived Phytosterol, Is an Antagonist of the Bile Acid Nuclear Receptor FXR}, volume={62}, ISSN={0031-3998 1530-0447}, url={http://dx.doi.org/10.1203/pdr.0b013e3181256492}, DOI={10.1203/pdr.0b013e3181256492}, abstractNote={Phytosterols, components of soy-derived lipids, are among the proposed exacerbants of parenteral nutrition-associated cholestasis (PNAC). We investigated whether phytosterols contribute to bile acid (BA)-induced hepatocyte damage by antagonizing a nuclear receptor (NR) critically involved in hepatoprotection from cholestasis, FXR (farnesoid X receptor, NR1H4). In HepG2 cells, stigmasterol acetate (StigAc), a water-soluble Stig derivative, suppressed ligand-activated expression of FXR target genes involved in adaptation to cholestasis (i.e. BSEP, FGF-19, OSTalpha/beta). Furthermore, StigAc antagonized BA-activated, FXR target genes SHP and BSEP in FXR+/+, but not in FXR-/- mouse hepatocytes. Both Stig and StigAc inhibited BA-activated, FXR-dependent reporter gene expression in transfected HepG2 cells, whereas the most prevalent phytosterol in lipids, beta-sitosterol, had no inhibitory effect. Finally, among six ligand-activated NR-ligand binding domains (LBDs) tested, antagonism by StigAc was specific to only two (FXR and PXR, pregnane X receptor, NR1I2). We demonstrate that Stig, a phytosterol prevalent in soy-derived PN lipid solutions, is a potent in vitro antagonist of the NR for bile acids FXR.}, number={3}, journal={Pediatric Research}, publisher={Springer Science and Business Media LLC}, author={Carter, Beth A and Taylor, Olga A and Prendergast, Daniel R and Zimmerman, Tracy L and Von Furstenberg, Richard and Moore, David D and Karpen, Saul J}, year={2007}, month={Sep}, pages={301–306} }
 @article{carter_taylor_von furstenberg_karpen_2006, title={Soy lipid-derived stigmasterol (STIG) antagonizes bile acid (BA)-activation of the FXR dependent BA sinusoidal efflux pump genes, OSTα/β, in 1° mouse hepatocytes and HEPG2 cells}, volume={43}, url={https://doi.org/10.1002/j.1536-4801.2006.tb13812.x}, DOI={10.1002/j.1536-4801.2006.tb13812.x}, abstractNote={Journal of Pediatric Gastroenterology and NutritionVolume 43, Issue 4 p. E35-E35 North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition Annual Meeting, October 19-22, 2006, Orlando, Florida: Abstracts: POSTER SESSION II, FRIDAY, OCTOBER 20, 2006, 12:15 p.m. - 2:15 p.m.: Hepatobiliary/Transport: 69 SOY LIPID-DERIVED STIGMASTEROL (STIG) ANTAGONIZES BILE ACID (BA)-ACTIVATION OF THE FXR DEPENDENT BA SINUSOIDAL EFFLUX PUMP GENES, OSTα/β, IN 1° MOUSE HEPATOCYTES AND HEPG2 CELLS Beth A. Carter, Beth A. Carter Baylor College of Medicine, Houston, TXSearch for more papers by this authorOlga A. Taylor, Olga A. Taylor Baylor College of Medicine, Houston, TXSearch for more papers by this authorRichard von Furstenberg, Richard von Furstenberg Baylor College of Medicine, Houston, TXSearch for more papers by this authorSaul J. Karpen, Saul J. Karpen Baylor College of Medicine, Houston, TXSearch for more papers by this author Beth A. Carter, Beth A. Carter Baylor College of Medicine, Houston, TXSearch for more papers by this authorOlga A. Taylor, Olga A. Taylor Baylor College of Medicine, Houston, TXSearch for more papers by this authorRichard von Furstenberg, Richard von Furstenberg Baylor College of Medicine, Houston, TXSearch for more papers by this authorSaul J. Karpen, Saul J. Karpen Baylor College of Medicine, Houston, TXSearch for more papers by this author First published: 01 October 2006 https://doi.org/10.1002/j.1536-4801.2006.tb13812.xRead the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume43, Issue4October 2006Pages E35-E35 RelatedInformation}, number={4}, journal={Journal of Pediatric Gastroenterology and Nutrition}, author={Carter, B.A. and Taylor, O.A. and von Furstenberg, R. and Karpen, S.J.}, year={2006}, month={Oct}, pages={E35} }
 @article{carter_prendergast_von furstenberg_karpen_2005, title={SOY‐LIPID DERIVED STIGMASTEROL SUPPRESSES FXR TARGET GENES BSEP AND FGF‐19 IN HUMAN HEPATOBLASTOMA (HEPG2) CELLS‐POTENTIAL ROLE IN TOTAL PARENTERAL NUTRITION‐ASSOCIATED CHOLESTASIS (TPNAC)}, volume={41}, ISSN={0277-2116 1536-4801}, url={http://dx.doi.org/10.1097/01.mpg.0000182046.11869.a2}, DOI={10.1097/01.mpg.0000182046.11869.a2}, abstractNote={Background: The etiology of TPNAC is unknown but may involve an impairment in bile acid (BA) secretion leading to intracellular accumulation of BAs. BSEP, the 1° bile salt exporter on the canalicular membrane of the hepatocyte, has an FXR element in its promoter upregulated by the potent FXR ligand chenodeoxycholic acid (CDCA). We previously reported that stigmasterol acetate (StigAc), a phytosterol component of soy lipid in IV fat solutions, antagonizes BA-activated BSEP in a 1° mouse hepatocyte model. Fibroblast growth factor-19 (FGF-19), like BSEP, has an FXR response element in its promoter and has previously been implicated in the suppression of BA biosynthesis (Holt et al, Genes and Dev, 2003). Purpose: To test the effect of stigmasterol on BA-activated FXR target genes in a human cell line. Methods: HepG2 cells were grown to 80% confluency followed by 24 hr treatments in 0.25% charcoal stripped media: Vehicle alone, chenodeoxycholic acid (CDCA) 100 μM alone, StigAc 10 μM alone, or CDCA 100 μM + StigAc 10 μM. Cells were harvested, RNA isolated, and QRTPCR performed to determine the relative expression of the human FXR target genes BSEP and FGF-19 normalized to 18s RNA expression. Results: As expected, the potent FXR ligand, CDCA, activated BSEP mRNA expression ~300-fold over vehicle-treated cells. StigAc alone had no effect on BA-activated HepG2 cells, while cells co-treated with CDCA + StigAc suppressed BA-activated BSEP mRNA by ~50%. StigAc similarly suppressed BA-activated FGF-19 expression in HepG2 cells (~70% suppression). Conclusion: The high serum levels of stigmasterol in infants with TPNAC may contribute to hepatotoxicity by interfering with the hepatoprotective pathways orchestrated by FXR (i.e. BA-induced upregulation of BSEP and FGF-19 mediated suppression of BA synthesis). These findings further delineate our molecular model of TPNAC in infants on longterm TPN.}, number={4}, journal={Journal of Pediatric Gastroenterology and Nutrition}, publisher={Wiley}, author={Carter, Beth A and Prendergast, Daniel R and von Furstenberg, Richard J and Karpen, Saul J}, year={2005}, month={Oct}, pages={552–552} }