TY - JOUR TI - HIF1alpha is essential for normal intrauterine differentiation of alveolar epithelium and surfactant production in the newborn lung of mice. AU - Saini, Y AU - LaPres, JJ T2 - The Journal of biological chemistry AB - Neonatal respiratory distress syndrome (RDS) is mainly the result of perturbation in surfactant production and is a common complication seen in premature infants. Normal fetal lung development and alveolar cell differentiation is regulated by a network of transcription factors. Functional loss of any of these factors will alter the developmental program and impact surfactant production and normal gas exchange. During development, the fetus is exposed to varying oxygen concentrations and must be able to quickly adapt to these changes in order to survive. Hypoxia-inducible factor 1α (HIF1α) is the primary transcription factor that is responsible for regulating the cellular response to changes in oxygen tension and is essential for normal development. Its role in lung maturation is not well defined and to address this knowledge gap, a lung-specific HIF1α knock-out model has been developed. Loss of HIF1α early in lung development leads to pups that die within hours of parturition, exhibiting symptoms similar to RDS. Lungs from these pups display impaired alveolar epithelial differentiation and an almost complete loss of surfactant protein expression. Ultrastructural analysis of lungs from HIF1α deletion pups had high levels of glycogen, aberrant septal development, and decreased expression of several factors necessary for proper lung development, including HIF2α, β-catenin, and vascular endothelial growth factor. These results suggest that HIF1α is essential for proper lung maturation and alteration in its normal signaling during premature delivery might explain the pathophysiology of neonatal RDS. Neonatal respiratory distress syndrome (RDS) is mainly the result of perturbation in surfactant production and is a common complication seen in premature infants. Normal fetal lung development and alveolar cell differentiation is regulated by a network of transcription factors. Functional loss of any of these factors will alter the developmental program and impact surfactant production and normal gas exchange. During development, the fetus is exposed to varying oxygen concentrations and must be able to quickly adapt to these changes in order to survive. Hypoxia-inducible factor 1α (HIF1α) is the primary transcription factor that is responsible for regulating the cellular response to changes in oxygen tension and is essential for normal development. Its role in lung maturation is not well defined and to address this knowledge gap, a lung-specific HIF1α knock-out model has been developed. Loss of HIF1α early in lung development leads to pups that die within hours of parturition, exhibiting symptoms similar to RDS. Lungs from these pups display impaired alveolar epithelial differentiation and an almost complete loss of surfactant protein expression. Ultrastructural analysis of lungs from HIF1α deletion pups had high levels of glycogen, aberrant septal development, and decreased expression of several factors necessary for proper lung development, including HIF2α, β-catenin, and vascular endothelial growth factor. These results suggest that HIF1α is essential for proper lung maturation and alteration in its normal signaling during premature delivery might explain the pathophysiology of neonatal RDS. During development, an embryo is exposed to varying levels of oxygen as a balance is created between vascularization and tissue growth. Localized hypoxia, a decrease in available oxygen, is a normal part of this process. The programmed responses to these decreases in available oxygen are essential for viability (1Webster W.S. Abela D. Birth Defects Res. C Embryo Today. 2007; 81: 215-228Crossref PubMed Scopus (132) Google Scholar, 2Giaccia A.J. Simon M.C. Johnson R. Genes Dev. 2004; 18: 2183-2194Crossref PubMed Scopus (286) Google Scholar). In utero, the embryo is supplied with oxygen and nutrients through the placental barrier. Following parturition, oxygen is supplied by the neonatal lungs and therefore, proper intrauterine development of the alveolar gas exchange regions of the lung is essential for the newborn's first breath and for sustaining life outside the womb (3James Metcalfe H.B. Waldemar MollPhysiol. Rev. 1967; 47: 782-838Crossref PubMed Scopus (77) Google Scholar). Lung morphogenesis is a complex process that is orchestrated by several transcription factors, growth factors, and extracellular cues (4Groenman F. Unger S. Post M. Biol. Neonate. 2005; 87: 164-177Crossref PubMed Scopus (67) Google Scholar). For example, thyroid transcription factor-1 regulates the expression of the genes for Clara cell secretory protein (CCSP) 2The abbreviations used are: CCSP, Clara cell secretory protein; CEBPα, CCAAT-enhancer-binding protein α; Foxa1, Forkhead box A1; Foxa2, Forkhead box A2; GATA 6, GATA-binding protein 6; HGPRT, hypoxanthine guanine phosphoribosyl transferase; HIF, hypoxia-inducible factor; HIMF, hypoxia-inducible mitotic factor; HNF3α, hepatocyte nuclear factor 3α; PAS, periodic acid-Schiff; qRT-PCR, quantitative real-time PCR; RDS, respiratory distress syndrome; rtTA, reverse tetracycline transactivator; SP(A-D), surfactant-associated proteins A-D; VEGF, vascular endothelial growth factor. 2The abbreviations used are: CCSP, Clara cell secretory protein; CEBPα, CCAAT-enhancer-binding protein α; Foxa1, Forkhead box A1; Foxa2, Forkhead box A2; GATA 6, GATA-binding protein 6; HGPRT, hypoxanthine guanine phosphoribosyl transferase; HIF, hypoxia-inducible factor; HIMF, hypoxia-inducible mitotic factor; HNF3α, hepatocyte nuclear factor 3α; PAS, periodic acid-Schiff; qRT-PCR, quantitative real-time PCR; RDS, respiratory distress syndrome; rtTA, reverse tetracycline transactivator; SP(A-D), surfactant-associated proteins A-D; VEGF, vascular endothelial growth factor. produced in Clara cells in the tracheobronchial airways and various surfactants produced by alveolar type II cells in the lung parenchyma (5Cassel T.N. Suske G. Nord M. Ann. N. Y. Acad. Sci. 2000; 923: 300-302Crossref PubMed Scopus (11) Google Scholar, 6deFelice M. Silberschmidt D. DiLauro R. Xu Y. Wert S.E. Weaver T.E. Bachurski C.J. Clark J.C. Whitsett J.A. J. Biol. Chem. 2003; 278: 35574-35583Abstract Full Text Full Text PDF PubMed Scopus (128) Google Scholar). CCAAT-enhancer-binding protein α (CEBPα) is essential for proper regulation of alveolar Type II cell differentiation, and Forkhead box A2 (Foxa2) controls various cellular programs involved in lung development (e.g. surfactant expression) (6deFelice M. Silberschmidt D. DiLauro R. Xu Y. Wert S.E. Weaver T.E. Bachurski C.J. Clark J.C. Whitsett J.A. J. Biol. Chem. 2003; 278: 35574-35583Abstract Full Text Full Text PDF PubMed Scopus (128) Google Scholar, 7Mucenski M.L. Wert S.E. Nation J.M. Loudy D.E. Huelsken J. Birchmeier W. Morrisey E.E. Whitsett J.A. J. Biol. Chem. 2003; 278: 40231-40238Abstract Full Text Full Text PDF PubMed Scopus (252) Google Scholar, 8Wan H.J. Xu Y. Ikegami M. Stahlman M.T. Kaestner K.H. Ang S.L. Whitsett J.A. Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 14449-14454Crossref PubMed Scopus (108) Google Scholar). Ablation of any of these factors results in major structural and functional abnormalities ranging from undeveloped alveolar structure and/or improper airway branching to the faulty processing of various secretory components (7Mucenski M.L. Wert S.E. Nation J.M. Loudy D.E. Huelsken J. Birchmeier W. Morrisey E.E. Whitsett J.A. J. Biol. Chem. 2003; 278: 40231-40238Abstract Full Text Full Text PDF PubMed Scopus (252) Google Scholar, 8Wan H.J. Xu Y. Ikegami M. Stahlman M.T. Kaestner K.H. Ang S.L. Whitsett J.A. Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 14449-14454Crossref PubMed Scopus (108) Google Scholar). One of the extracellular cues that is important for fetal vascular growth and lung morphogenesis is the physiologically low O2 environment of the fetus. The ability to cope with this developmental “hypoxia” is not only important for lung maturation but essential for the viability of the fetus (1Webster W.S. Abela D. Birth Defects Res. C Embryo Today. 2007; 81: 215-228Crossref PubMed Scopus (132) Google Scholar, 2Giaccia A.J. Simon M.C. Johnson R. Genes Dev. 2004; 18: 2183-2194Crossref PubMed Scopus (286) Google Scholar). Cellular responses to decreased oxygen availability are regulated by a family of proteins called hypoxia-inducible factors (HIFs). The ability of HIFs to respond to hypoxia is controlled by oxygen-dependent post-translational hydroxylation. The prolyl hydroxylases, PHDs, modify HIFs on essential residues in an oxygen-, iron-, and α-ketoglutarate-dependent manner. Once hydroxylated, the HIF is quickly degraded in a proteosomal-dependent process that involves the Von Hippel Lindau (VHL) tumor suppressor. HIF1α, the most ubiquitously expressed HIF, has been shown to play a critical role in normal development. HIF1α regulates the expression of genes important for cellular adaptation to hypoxia, including glycolytic enzymes and angiogenic factors (9Semenza G.L. J. Appl. Physiol. 2000; 88: 1474-1480Crossref PubMed Scopus (1461) Google Scholar). Gene deletion studies have established that HIF1α is indispensable during fetal development as HIF1α-/- mice die at midgestation, in association with defects in VEGF expression and vascularization (10Ryan H.E. Lo J. Johnson R.S. EMBO J. 1998; 17: 3005-3015Crossref PubMed Scopus (1318) Google Scholar, 11Kotch L.E. Iyer N.V. Laughner E. Semenza G.L. Dev. Biol. 1999; 209: 254-267Crossref PubMed Scopus (333) Google Scholar, 12Iyer N.V. Kotch L.E. Agani F. Leung S.W. Laughner E. Wenger R.H. Gassmann M. Gearhart J.D. Lawler A.M. Yu A.Y. Semenza G.L. Gene Dev. 1998; 12: 149-162Crossref PubMed Scopus (2008) Google Scholar). In addition, HIF1+/- mice developed pulmonary hypertension and pulmonary vascular remodeling under hypoxic conditions (13Yu A.Y. Shimoda L.A. Iyer N.V. Huso D.L. Sun X. McWilliams R. Beaty T. Sham J.S. Wiener C.M. Sylvester J.T. Semenza G.L. J. Clin. Investig. 1999; 103: 691-696Crossref PubMed Scopus (554) Google Scholar). PHD inhibitor-induced HIF stabilization has been shown to improve lung growth in prematurely born baboon neonates, predominantly through enhanced expression of VEGF (14Asikainen T.M. Chang L.Y. Coalson J.J. Schneider B.K. Waleh N.S. Ikegami M. Shannon J.M. Winter V.T. Grubb P. Clyman R.I. Yoder B.A. Crapo J.D. White C.W. FASEB J. 2006; 20: 1698-1700Crossref PubMed Scopus (63) Google Scholar). Finally, loss of HIF2α perturbs normal lung development through loss of VEGF expression and to subsequently decrease alveolar type II cell production of pulmonary surfactants (15Compernolle V. Brusselmans K. Acker T. Hoet P. Tjwa M. Beck H. Plaisance S. Dor Y. Keshet E. Lupu F. Nemery B. Dewerchin M. Van Veldhoven P. Plate K. Moons L. Collen D. Carmeliet P. Nat. Med. 2002; 8: 702-710Crossref PubMed Google Scholar). Previous studies using systemic HIF1α deletions have demonstrated its central role in development and partial HIF1α deletions resulted in disrupted normal functioning of tissues such as heart and lung (13Yu A.Y. Shimoda L.A. Iyer N.V. Huso D.L. Sun X. McWilliams R. Beaty T. Sham J.S. Wiener C.M. Sylvester J.T. Semenza G.L. J. Clin. Investig. 1999; 103: 691-696Crossref PubMed Scopus (554) Google Scholar, 16Tomita S. Ueno M. Sakamoto M. Kitahama Y. Ueki M. Maekawa N. Sakamoto H. Gassmann M. Kageyama R. Ueda N. Gonzalez F.J. Takahama Y. Mol. Cell Biol. 2003; 23: 6739-6749Crossref PubMed Scopus (200) Google Scholar, 17Ryan H.E. Poloni M. McNulty W. Elson D. Gassmann M. Arbeit J.M. Johnson R.S. Cancer Res. 2000; 60: 4010-4015PubMed Google Scholar). The present study was aimed at elucidating the role(s) of HIF1α in the in utero differentiation of the alveolar epithelium. To this aim, mice with a lung-specific deletion of HIF1α (termed HIF1αΔ/Δ) were generated. Lung specific embryonic deletion of HIF1α earlier than 3 days prior to parturition led to HIF1αΔ/Δ neonates exhibiting cyanosis and respiratory failure soon after birth. The expression of surfactant proteins was significantly lower in HIF1αΔ/Δ pups as compared with littermate control pups. Examination of the lungs from HIF1αΔ/Δ pups via light and transmission electron microscopy confirmed defects in both alveolar epithelial differentiation and septal development that resulted in the observed respiratory distress. Transgenic Mice and Genotyping—HIF1αflox/flox and SP-C-rtTA-/tg/(tetO)7-CMV-Cretg/tg transgenic mice were generous gifts from Randall Johnson (University of California, San Diego) and Jeffrey Whitsett (Cincinnati Children's Hospital Medical Center), respectively (10Ryan H.E. Lo J. Johnson R.S. EMBO J. 1998; 17: 3005-3015Crossref PubMed Scopus (1318) Google Scholar, 17Ryan H.E. Poloni M. McNulty W. Elson D. Gassmann M. Arbeit J.M. Johnson R.S. Cancer Res. 2000; 60: 4010-4015PubMed Google Scholar, 19Lobe C.G. Koop K.E. Kreppner W. Lomeli H. Gertsenstein M. Nagy A. Dev. Biol. 1999; 208: 281-292Crossref PubMed Scopus (441) Google Scholar). HIF1αflox/flox and SP-C-rtTA-/tg/(tetO)7-CMV-Cretg/tg transgenic mice were mated to generate SP-C-rtTA-/tg/(tetO)7-CMV-Cretg/tg/HIF1αflox/flox mice capable of respiratory epithelium-specific conditional recombination in the floxed HIF1α gene. In this model, depending on the day doxycycline is administered to the dam, various cell populations within the lung undergo recombination in HIF1αflox/flox alleles (20Bradford M.M. Anal. Biochem. 1976; 72: 248-254Crossref PubMed Scopus (211983) Google Scholar). Genotyping of the mice progeny was performed by PCR for all the three loci using previously published primer sequences (supplemental Table S1). Genomic DNA extraction from tail clipping was performed using the Direct PCR extraction system (Viagen Biotech) per the manufacturer's instructions. PCR conditions were standardized for all the three alleles: denaturation at 94 °C for 3 min; 38 cycles of denaturation at 94 °C for 45 s, annealing at 60 °C for 45 s, and polymerization at 72 °C for 60 s followed by a 7-min extension at 72 °C. Sizes of the amplified products obtained were ∼210 bp for HIF1α (wild type); 244 bp for HIF1αflox/flox; 370 bp for Cre transgene; and 350 bp for rtTA transgene (Fig. 1B). Doxycycline Treatment and Animal Husbandry—Dams bearing triple (SP-C-rtTA-/tg/(tetO)7-CMV-Cretg/tg/HIF1αflox/flox) and double ((tetO)7-CMV-Cretg/tg/HIF1αflox/flox) transgenic embryos were maintained on doxycycline feed (625 mg of doxycycline/kg; Harlan Teklad, Madison, WI) and drinking water (0.8 mg/ml; Sigma Chemicals Co.). The day of parturition was taken as the reference point to calculate the duration of doxycycline exposure. Control and HIF1aΔ/Δ pups exposed to doxycycline treatment for 0-14 days in utero were sacked within 1 h of parturition. All pups were sacrificed by decapitation. Mice used in this study were kept at the animal housing facility under the strict hygienic and pathogen-free conditions approved by the University Laboratory Animal Resource regulatory unit. Lung Harvesting and Processing—3-8 pups were analyzed from each genotype and doxycycline treatment. Pups were assessed for total body and lung weight. Lung tissues were isolated and the left lung lobe was fixed in 10% neutral-buffered formalin (NBF) or 4% glutaraldehyde for morphological analysis. The remaining lung tissue was divided; half was snap-frozen for Western blotting, half was stored in RNAlater RNA Stabilization Reagent (Qiagen) for RNA isolation and qRT-PCR analysis. Western Blotting—Snap-frozen lung tissue (10 mg) was lysed and homogenized in radioimmune precipitation assay buffer (50 mm Tris-HCl, 1% Nonidet P-40, 0.25% sodium deoxycholate, 150 mm NaCl, 1 mm each of EDTA, phenylmethylsulfonyl fluoride, sodium vandate, sodium fluoride, 1 m dithiothreitol) containing protease inhibitors (1 μg/ml each of aprotinin, leupeptin, pepstatin) using a bead beater system (45 s at 30 Hz frequency). Insoluble material was removed by centrifugation (10,000 × g for 10 min.) and protein concentration was measured in supernatant using Bradford protein assay (20Bradford M.M. Anal. Biochem. 1976; 72: 248-254Crossref PubMed Scopus (211983) Google Scholar). Protein samples (30 μg) were separated by SDS-PAGE (NuPage 4-12% Bis-Tris gradient gel, Invitrogen) and transferred to a nitrocellulose membrane. Western blots were performed with rabbit or goat antibodies against surfactant-associated proteins (SP-A (sc 13977, Santa Cruz Biotechnology), SP-B (AB 3780, Chemicon), SP-C (sc-13979, Santa Cruz Biotechnology) and SP-D (SC 7709, Santa Cruz Biotechnology), HIF1α (NB100-479, Novus Biologicals), tubulin (T5168, Sigma), and β-actin (SC-7210, Santa Cruz Biotechnology). Proteins were visualized with horseradish peroxidase-conjugated goat anti-rabbit IgG (sc-2004, Santa Cruz Biotechnology) or rabbit anti-goat (sc-2768, Santa Cruz Biotechnology) and ECL Western blot system (Pierce) or fluorescently using Alexa fluor 680 anti-rabbit IgG (A21109, Invitrogen) or IRdye800 antimouse IgG (610-132-121, Rockland). Histopathology and Immunohistochemistry—At least 4-6 pups of each genotype and doxycycline treatment were analyzed for histopathological changes. Formalin-fixed left lung lobe tissues were paraffin-embedded and 5-micron thick sections were mounted on glass slides and stained with hematoxylin and eosin (H&E), periodic acid Schiff (PAS), or immunostained with SP-B (1:500 dilution, AB3780, Chemicon, MA), SP-C (1:4000 dilution, antibody kindly provided by Jeffrey Whitsett), and CRE (1:100 dilution, AB24608; Abcam, Cambridge, MA). Briefly, tissue sections were deparaffinized and endogenous peroxidase activity was quenched by incubation with 6% H2O2 for 30 min. Immunostaining was performed using Rabbit Vector Elite ABC kit (Vector Laboratories), according to the manufacturer's recommendations. Transmission Electron Microscopy—Neonatal lung sections were fixed in 4% buffered glutaraldehyde overnight at 4 °C, and stored in the 10% NBF fixative until postfixation with 1% phosphate-buffered osmium tetroxide. Tissues were dehydrated through a graded series of ethanol and propylene oxide, and embedded in Poly/Bed-Araldite resin (Polysciences, Inc., Warrington, PA). 1 μm-thick sections were cut and stained with toluidine blue for light microscopic identification of specific tissue sites for transmission electron microscopy (TEM). Ultrathin tissue sections for TEM were cut at ∼75 nm and stained with lead citrate and uranyl acetate. Sectioning was performed with an LKB Ultratome III (LKB Instruments, Inc., Rockville, MD). Ultrastructural tissue examination and photography were performed with a JEOL JEM 100CXII electron microscope (JEOL Ltd., Tokyo, Japan). RNA Isolation and Quantitative Real-time PCR (qRT-PCR) Analysis—10 mg of lung tissue stored in RNAlater RNA Stabilization Reagent was homogenized in RLT buffer using a Retsch MM200 bead beater system (Retsch, Haan, Germany). Total RNA quantification was performed spectrophotometrically (NanoDrop ND-1000 UV-Vis Spectrophotometer). Total RNA (1 μg) was reverse-transcribed using the Superscript II reverse transcriptase kit (Invitrogen). The expression level of selected genes involved in surfactant metabolism and lung development was analyzed by qRT-PCR using SYBR green (Applied Biosystems, Foster City, CA) as previously described (21Vengellur A. LaPres J.J. Toxicol. Sci. 2004; 82: 638-646Crossref PubMed Scopus (120) Google Scholar). Gene-specific primers are listed in supplemental Table S2. Copy number was determined by comparison with standard curves of the respective genes. This measurement was controlled for RNA quality, quantity, and RT efficiency by normalizing it to the expression level of the hypoxanthine guanine phosphoribosyl transferase (HGPRT) gene. Statistical significance was determined by use of normalized relative changes and analysis of variance. Quantitative Analysis—qRT-PCR analysis was performed using unpaired two-tailed Student's t test on GraphPad Prism. Results were considered significant at the 5% level. Conditional Inactivation of HIF1α in the Lung—The HIF1α gene was inactivated in the lung by mating HIF1α-conditional null mice (HIF1αflox/flox) (10Ryan H.E. Lo J. Johnson R.S. EMBO J. 1998; 17: 3005-3015Crossref PubMed Scopus (1318) Google Scholar) to an inducible bitransgenic mouse, SPC-rtTA, that expresses the reverse tetracycline transactivator (rtTA) under the control of the human surfactant protein C (SP-C) promoter and the Cre recombinase gene with a tetracycline operon (Fig. 1A) (18Perl A.K. Wert S.E. Nagy A. Lobe C.G. Whitsett J.A. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: 10482-10487Crossref PubMed Scopus (400) Google Scholar). SPC-rtTA mice have been shown to express Cre recombinase specifically in the epithelial cells of the primordial lung buds as well as in the alveolar and bronchiolar epithelium of postnatal mice in the presence of an inducer (i.e. doxycycline) (19Lobe C.G. Koop K.E. Kreppner W. Lomeli H. Gertsenstein M. Nagy A. Dev. Biol. 1999; 208: 281-292Crossref PubMed Scopus (441) Google Scholar). In the present study, compound litters from reciprocal matings between SP-C-rtTA-/tg/(tetO)7-CMV-Cretg/tg/HIF1αflox/flox and (tetO)7-CMV-Cretg/tg/HIF1αflox/flox mice were composed of pups with mixed genotypes with respect to SPC-rtTA transgene. Their genotypic ratios (data not shown) were in accordance with Mendelian inheritance as demonstrated by genotyping (Fig. 1B). Analysis of these ratios from litters studied (n = 30 litters and 190 pups) confirmed no in utero mortalities (data not shown). In the absence of doxycycline treatment, pups were phenotypically indistinguishable between litters. The functionality of the doxycycline treatment was demonstrated by the significant decrease in HIF1α mRNA as measured by qRT-PCR (Fig. 1C) and loss of HIF1α protein by Western blot analysis (Fig. 1D). Lung-specific Deletion of HIF1α Causes Lethal Phenotype—To determine if in utero loss of HIF1α influenced litter size or the Mendelian distribution of pups, dams carrying triple transgenic embryos were exposed to doxycycline through feed and water for various lengths of time prior to parturition (i.e. 2, 4, 6, 8, 10, 12, and 14 days). Our findings indicate pups exposed to doxycycline in utero for at least 8 days prior to parturition had a 100% mortality rate, while those receiving the drug for 4-6 days prior to parturition had approximately a 15% chance of survival (Fig. 2A). Following parturition, HIF1αΔ/Δ pups (doxycycline-induced) developed respiratory distress, severe cyanosis, and died within an hour of birth (range of 10-60 min) (Fig. 2B). No observable differences were noted in survival times following parturition from pups treated with doxycycline for only 1-6 days compared with those exposed for at least 8 days. More importantly, no mortality was observed in HIF1α+fl/+fl mice lacking either the rtTA or Cre recombinase transgene in the presence or absence of doxycycline (data not shown). These data suggest that the mortality is due to loss of HIF1α and not off-target effects of the other transgenes. At necropsy, the lungs from HIF1αΔ/Δ pups were dark red and atelectic, in contrast to pink, aerated lungs from control (in the absence of doxycycline treatment) pups. Excised lung lobes from HIF1αΔ/Δ pups weighed slightly more than the lungs from control mice and sank to the bottom of a saline-filled vial in contrast to the lungs from control littermates, which floated in saline (Fig. 2, C and D). At birth, the average total body weights of control littermate and HIF1αΔ/Δ were 1.318±0.018 (S.E.) and 1.288 ± 0.024 grams, respectively. Defective Lung Morphology—The lungs of HIF1αΔ/Δ pups had histologic and ultrastructural features that indicate an impairment of normal in utero differentiation of alveolar epithelium, with a related loss of lung surfactant proteins that are essential for preventing collapse of individual alveoli and ensuring proper gas exchange to maintain life. Lungs of HIF1αΔ/Δ pups at birth had abnormally thickened alveolar septa (Fig. 3C) lined by undifferentiated cuboidal epithelial cells (immature pneumocytes) containing large amounts of intracytoplasmic periodic acid Schiff (PAS)-positive material (Fig. 3D) that was ultrastructurally identified, via transmission electron microscopy, as glycogen (Fig. 4, C and D). There was a marked reduction of alveolar airspaces in the lungs of these mice due in part to the thickened septa and to focal areas of atelectasis.FIGURE 4Electron photomicrographs of alveolar epithelium of newborn pups. The lung from the control pup (A and B) has a well differentiated alveolar epithelium containing both type I and II cells (B). Stippled arrow indicates thin, squamoid, type I cells, and solid arrows indicate tubular myelin figures (secreted surfactant) within the alveolar airspace of control pup. Type II cells have numerous intracytoplasmic lamellar bodies (lb) and apical microvilli (mv). In contrast the alveolar surface in HIF1αΔ/Δ pup (C and D) is lined by large undifferentiated cuboidal epithelial cells. The cytoplasm of these epithelial cells are primarily filled with glycogen (*). No lamellar bodies are present in these cells. c, capillary; rbc, red blood cells; II, type II alveolar epithelial cell; I, type I alveolar epithelial cell; if, interstitial fibroblast; *, glycogen; n, nucleus; 1, mitochondrion; 2, rough endoplasmic reticulum; pl, pleural surface; solid black arrows, tubular myelin; stippled arrow, type I cells. All doxycycline treatments were at least 12 days.View Large Image Figure ViewerDownload Hi-res image Download (PPT) In contrast, the lungs of control mice at birth had uniformly dilated alveolar airspaces with much thinner alveolar septa that were lined by more differentiated surface epithelium containing much less PAS-positive glycogen (Fig. 3, A and B). Ultrastructurally, the luminal epithelium lining the alveolar airspace of control pups consisted of both alveolar type I and II cells (differentiated pneumocytes). Distinctive lamellar bodies were present in the apical cytoplasm of alveolar type II cells along with tubular myelin and secreted lamellar material (ultrastructural features of secreted surfactant) widely scattered along the alveolar luminal surface (Fig. 4, A and B). These distinctive cuboidal epithelial cells also had a round to ovoid nucleus, an apical luminal surface lined by short microvilli, and numerous cytoplasmic organelles consisting of mitochondria, rough endoplasmic reticulum, and lysosomes. Type I cells were identified by their thin squamous morphology containing few organelles, and a fusiform nucleus (Fig. 4, A and B). Interestingly, there were no microscopically detectable differences in the morphology of intrapulmonary conducting airways (preterminal and terminal bronchioles), lined principally by ciliated cells and nonciliated cuboidal (Clara) cells, between control and HIF1αΔ/Δ pups at birth. Altered Surfactant Metabolism—qRT-PCR analysis of the genes for surfactant proteins show a decreased expression of SP-A, SP-B, and SP-C in the HIF1αΔ/Δ mice compared with control pups (Fig. 5A). SP-D mRNA expression was not significantly different between the two genotypes. Western blot analysis of protein extract from HIF1αΔ/Δ and control animals was also performed to determine if surfactant protein levels were similar to gene expression patterns. All four surfactants displayed a substantial reduction in protein levels in the HIF1αΔ/Δ mice as compared with control pups (Fig. 5B). Interestingly, SP-C displayed an overall decrease in protein levels but the pre-processed form (Fig. 5B, SPC upper band) was not as affected by deletion of HIF1α as was the fully processed form (Fig. 5B, SPC lower band). This suggests that SP-C transcription is being decreased by the loss of HIF1α and processing of any translated SP-C protein is being inhibited by the aberrant differentiation of the HIF1αΔ/Δ lung. In comparison, SP-B monomer and dimer forms were equally affected by loss of HIF1α. Immunohistochemical staining on lung sections confirmed the reduced levels of SP-B and SP-C in the HIF1αΔ/Δ compared with control pups (Fig. 5C). The reduced levels of surfactant mRNA and protein expression were consistent with the lack of alveolar epithelial differentiation and respiratory distress in the HIF1αΔ/Δ pups. Altered Gene Expression—To begin to understand how loss of HIF1α induced changes in lung morphology, qRT-PCR was performed on several critical genes known to be involved in lung development. These genes include β-catenin, Clara cell secretory protein (CCSP), forkhead box A2 (Foxa2), forkhead box protein A1 (Foxa1, also known as HNF3α), thyroid transcription factor 1 (TTF-1), GATA-binding protein 6 (Gata6), ATP-binding cassette, subfamily A (ABC1 member 3 (ABCA3), and CCAAT/enhancer-binding protein α (CEBPα). There were no significant differences in the expression levels of Foxa2, ABCA3, CEBPα, TTF-1, Foxa1, or GATA, suggesting that HIF1α is not required for their normal expression and is most likely involved in later events in lung development (Fig. 6 and data not shown). However, lower levels of β-catenin, CCSP, HIMF, VEGF, and HIF2α mRNA in neonatal HIF1αΔ/Δ pups compared with control littermates suggest that HIF1α plays a role in controlling their expression during maturation of the lung (7Mucenski M.L. Wert S.E. Nation J.M. Loudy D.E. Huelsken J. Birchmeier W. Morrisey E.E. Whitsett J.A. J. Biol. Chem. 2003; 278: 40231-40238Abstract Full Text Full Text PDF PubMed Scopus (252) Google Scholar, 15Compernolle V. Brusselmans K. Acker T. Hoet P. Tjwa M. Beck H. Plaisance S. Dor Y. Keshet E. Lupu F. Nemery B. Dewerchin M. Van Veldhoven P. Plate K. Moons L. Collen D. Carmeliet P. Nat. Med. 2002; 8: 702-710Crossref PubMed Google Scholar). Given the established role of these factors in regulating lung development, our data suggest that HIF1α is centrally located in the transcriptional network that is necessary for proper lung morphogenesis. The previously described role of HIF2α in lung development, the similarity in phenotype between the HIF2α knock-out survivors and HIF1α lung deletion strains, and the loss of HIF2α expression in the HIF1αΔ/Δ pups led us to determine the level of HIF2α expression at the protein level using immunohistochemistry (15Compernolle V. Brusselmans K. Acker T. Hoet P. Tjwa M. Beck H. Plaisance S. Dor Y. Keshet E. Lupu F. Nemery B. Dewerchin M. Van Veldhoven P. Plate K. Moons L. Collen D. Carmeliet P. Nat. Med. 2002; 8: 702-710Crossref PubMed Google Scholar). In the presence of doxycycline there was a pronounced expression of Cre recombinase in all cell types of the lung (Fig. 7D). There was a corresponding loss in HIF1α expression in the alveolar tissue (Fig. 7E). Though HIF1α is not expressed ubiquitously throughout the lung, its cell-specific expression was abrogated upon doxycycline treatment in the triple transgenic animal (Fig. 7). Surprisingly, the increased Cre recombinase and subsequent loss of HIF1α expression also led to a drastic decline in HIF2α levels in the lung (Fig. 7, C and F). Mammalian lung development and successful transition to extrauterine respiration involves a wide battery of biochemical, cellular, and ultrastructural changes. This complex process begins at embryonic day E9.5 with most of the alveolar epithelial differentiation events starting at embryonic day E17.5. Several transcription factors control the temporal and positional epithelial differentiation process, as well as the function of these differentiated cells. When the function of these critical factors is disrupted during development, lung function is compromised, leading to respiratory distress at birth. Our results have added HIF1α to this list of transcription factors that are critical for lung development. We have also demonstrated that loss of HIF1α during development leads to changes in the expression of other critical factors, such as HIF2α, VEGF, and β-catenin (23Maeda Y. Dave V. Whitsett J.A. Physiol. Rev. 2007; 87: 219-244Crossref PubMed Scopus (354) Google Scholar). HIFs are a class of transcription factors that play a critical role in oxygen-sensing and the metabolic adaptations to hypoxia. Previous studies have shown that HIF1α levels are predominant during lung development until day E13.5, at which time, HIF2α becomes the major HIF in this tissue (24Ema M. Taya S. Yokotani N. Sogawa K. Matsuda Y. FujiiKuriyama Y. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 4273-4278Crossref PubMed Scopus (833) Google Scholar, 25Shimoda L.A. Manalo D.J. Sham J.S.K. Semenza G.L. Sylvester J.T. Am. J. Physiol.-Lung C. 2001; 281: L202-L208Crossref PubMed Google Scholar). Targeted gene disruption of the HIF1α locus results in the embryonic lethality due to cardiovascular defects (12Iyer N.V. Kotch L.E. Agani F. Leung S.W. Laughner E. Wenger R.H. Gassmann M. Gearhart J.D. Lawler A.M. Yu A.Y. Semenza G.L. Gene Dev. 1998; 12: 149-162Crossref PubMed Scopus (2008) Google Scholar). Mice partially deficient for HIF1α alleles showed normal development but were physiologically compromised as they exhibited impaired pulmonary vascular remodeling under chronic hypoxic stress. In addition, the heterozygote was used to show that partial loss of HIF1α alleles results in impaired pulmonary arterial myocyte electrophysiological responses under hypoxic stress (13Yu A.Y. Shimoda L.A. Iyer N.V. Huso D.L. Sun X. McWilliams R. Beaty T. Sham J.S. Wiener C.M. Sylvester J.T. Semenza G.L. J. Clin. Investig. 1999; 103: 691-696Crossref PubMed Scopus (554) Google Scholar, 25Shimoda L.A. Manalo D.J. Sham J.S.K. Semenza G.L. Sylvester J.T. Am. J. Physiol.-Lung C. 2001; 281: L202-L208Crossref PubMed Google Scholar, 26Semenza G.L. Proc. Amer. Thor. Soc. 2005; 2: 68-70Crossref PubMed Scopus (41) Google Scholar). The study presented here has shown that the lung-specific loss of HIF1α leads to neonatal respiratory distress syndrome due to impaired alveolar epithelial differentiation. Interestingly, the lungs of HIF1αΔ/Δ pups had normal morphogenesis of conducting airways, with phenotypic alterations restricted to the alveolar parenchyma. These results suggest that HIF1α is involved in the late events of lung morphogenesis and alveolar epithelial differentiation but not in the early lung biogenesis and airway branching morphogenesis. Respiratory distress syndrome and associated histopathology are phenotypic hallmarks of targeted deletion of several transcription factors, such as Foxa2, CEBPα, GATA-6, SMAD3, calcineurin β1, and thyroid transcription factor-1, suggesting an intricate network of transcriptional regulation governing the complex ultrastructural maturation of lungs (8Wan H.J. Xu Y. Ikegami M. Stahlman M.T. Kaestner K.H. Ang S.L. Whitsett J.A. Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 14449-14454Crossref PubMed Scopus (108) Google Scholar, 22Martis P.C. Whitsett J.A. Xu Y. Perl A.K.T. Wan H.J. Ikegami M. Development. 2006; 133: 1155-1164Crossref PubMed Scopus (109) Google Scholar, 27Dave V. Childs T. Xu Y. Ikegami M. Besnard V. Maeda Y. Wert S.E. Neilson J.R. Crabtree G.R. Whitsett J.A. J. Clin. Investig. 2006; 116: 2597-2609Crossref PubMed Google Scholar, 28Devriendt K. Vanhole C. Matthijs G. de Zegher F. N. Engl. J. Med. 1998; 338: 1317-1318Crossref PubMed Scopus (196) Google Scholar, 29Liu C. Morrisey E.E. Whitsett J.A. Am. J. Physiol.-Lung C. 2002; 283: L468-L475Crossref PubMed Scopus (60) Google Scholar). Expression analysis demonstrated that the levels of Foxa2, Foxa1, CEBPα, and GATA-6 are comparable among pups with and without HIF1α (Fig. 6 and data not shown). This suggests that HIF1α acts either downstream or independent of these transcription factors. In contrast, the loss of HIF1α led to a decreased expression of other critical factors, including CCSP, β-catenin, HIMF, VEGF, and HIF2α (Fig. 6). Recently, the loss of HIF2α and subsequent decrease in VEGF has been linked to respiratory distress syndrome (15Compernolle V. Brusselmans K. Acker T. Hoet P. Tjwa M. Beck H. Plaisance S. Dor Y. Keshet E. Lupu F. Nemery B. Dewerchin M. Van Veldhoven P. Plate K. Moons L. Collen D. Carmeliet P. Nat. Med. 2002; 8: 702-710Crossref PubMed Google Scholar). These experiments involved a subset of HIF2α-/- mice that were viable up to parturition and displayed similar pathology to the HIF1αΔ/Δ mice described here. Delivery of VEGF, either intrauterine or postnatal intratracheal instillation, protected the HIF2α-/- mice from symptoms of respiratory distress syndrome. Given that HIF2α expression is abrogated in the HIF1aΔ/Δ pups suggests that HIF1α regulates HIF2α early in lung development and the subsequent loss of VEGF might explain the pathology of the HIF1αΔ/Δ pups. It remains to be seen if the decrease in VEGF expression is due to direct loss of HIF1α or indirectly through the loss of HIF2α expression. More importantly, the pathophysiology of the HIF1αΔ/Δ mouse resembles that of the HIF2α-/- mouse in relation to the lack of differentiation of type II pneumocytes and increased septal thickness. In addition, the HIF1αΔ/Δ pups display large glycogen stores with concurrent loss of surfactant protein expression. In conclusion, these studies provide evidence that HIF1α plays an important role in lung development. The respiratory failure induced by lung-specific deletion of HIF1α, suggests that hypoxia and HIF1α-regulated genes play an essential role in lung maturation, especially proper differentiation of type II alveolar cells. HIF1α potential upstream regulatory function of HIF2α and VEGF expression, directly or indirectly, offers a mechanism of action for the phenotypic observations. Finally, the results suggest that the hypoxia signaling cascade is important during the later stages of lung development but not essential in the initial patterning of the tissue. HIFs seem to play a later role in directing cell differentiation and regulating growth factors necessary for lung remodeling required for sustaining life extrauterine. Understanding the pathways necessary for proper HIF expression and function and the downstream pathways influenced by these factors will impact our ability to treat RDS and other diseases of the lung. We thank Dr. Randall Johnson (University of California, San Diego) for the gift of the conditional HIF1α mice and the gift of SP-C-rtTA/(tetO)7-CMV-Cre transgenic from Dr. Jeffrey Whitsett (Cincinnati Children's Hospital Medical Center). We would also like to acknowledge Melinda Kochenderfer for editorial help in preparing this manuscript. Download .pdf (.03 MB) Help with pdf files DA - 2008/9// PY - 2008/9// DO - 10.1074/jbc.m805927200 VL - 11 UR - http://europepmc.org/abstract/med/18801745 ER - TY - CONF TI - Modeling the effect of mixture components on the skin penetration AU - Ghafourian, T. AU - Samaros, E. AU - Riviere, J.E. T2 - 9th Annual Skin Forum C2 - 2008/// C3 - Proceedings of 9th Annual Skin Forum CY - London DA - 2008/// PY - 2008/// SP - 54 ER - TY - JOUR TI - Particulate matter in cigarette smoke alters iron homeostasis to produce a biological effect. T2 - American journal of respiratory and critical care medicine AB - Lung injury after cigarette smoking is related to particle retention. Iron accumulates with the deposition of these particles.We tested the postulate that (1) injury after smoking correlates with exposure to the particulate fraction of cigarette smoke, (2) these particles alter iron homeostasis, triggering metal accumulation, and (3) this alteration in iron homeostasis affects oxidative stress and inflammation.Rats and human respiratory epithelial cells were exposed to cigarette smoke, filtered cigarette smoke, and cigarette smoke condensate (the particulate fraction of smoke), and indices of iron homeostasis, oxidative stress, and inflammatory injury were determined. Comparable measures were also evaluated in nonsmokers and smokers.After exposure of rats to cigarette smoke, increased lavage concentrations of iron and ferritin, serum ferritin levels, and nonheme iron concentrations in the lung and liver tissue all increased. Lavage ascorbate concentrations were decreased, supporting an oxidative stress. After filtering of the cigarette smoke to remove particles, most of these changes were reversed. Exposure of cultured respiratory epithelial cells to cigarette smoke condensate caused a similar accumulation of iron, metal-dependent oxidative stress, and increased IL-8 release. Lavage samples in healthy smokers and smoking patients with chronic obstructive pulmonary disease revealed elevated concentrations of both iron and ferritin relative to healthy nonsmokers. Lavage ascorbate decreased with cigarette smoking. Serum iron and ferritin levels among smokers were increased, supporting systemic accumulation of this metal after cigarette smoke exposure.We conclude that cigarette smoke particles alter iron homeostasis, both in the lung and systemically. DA - 2008/8/21/ PY - 2008/8/21/ DO - 10.1164/rccm.200802-334oc UR - https://doi.org/10.1164/rccm.200802-334OC KW - smoking KW - ferritin KW - oxidants KW - chronic obstructive pulmonary disease ER - TY - JOUR TI - Molecular comparison of Mycobacterium avium isolates from clinical and environmental sources. T2 - Applied and environmental microbiology AB - We collected Mycobacterium avium isolates from clinical and drinking-water sources and compared isolates among themselves and to each other using molecular methods. Four clinical isolates were related to water isolates. Groups of indistinguishable clinical isolates were identified. The groups of identical clinical isolates suggest a common source of exposure. DA - 2008/6/6/ PY - 2008/6/6/ DO - 10.1128/aem.02900-07 UR - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/18539815/?tool=EBI ER - TY - JOUR TI - Human Health Effects Workgroup report. T2 - Advances in experimental medicine and biology DA - 2008/1/1/ PY - 2008/1/1/ DO - 10.1007/978-0-387-75865-7_26 UR - https://doi.org/10.1007/978-0-387-75865-7_26 ER - TY - JOUR TI - Ecosystem Effects Workgroup report. T2 - Advances in experimental medicine and biology DA - 2008/1/1/ PY - 2008/1/1/ DO - 10.1007/978-0-387-75865-7_31 UR - https://doi.org/10.1007/978-0-387-75865-7_31 ER - TY - CONF TI - Should physiological-based pharmacokinetic models become de riguer for veterinary antimicrobial studies? AU - Riviere, J.E. T2 - 4th International Conference on Antimicrobial Agents in Veterinary Medicine C2 - 2008/8/24/ C3 - Book of Abstracts: International Conference on Antimicrobial Agents in Veterinary Medicine CY - Prague, Czech Republic DA - 2008/8/24/ PY - 2008/8/24/ SP - 27 ER - TY - JOUR TI - Growth changes associated with feeding Clarias gariepinus with mouldy feeds AU - Aworh, M K AU - Alaka, O O AU - Olukunle, O AU - Olabode, H O T2 - Animal Production Research Advances DA - 2008/1/15/ PY - 2008/1/15/ DO - 10.4314/apra.v3i3.36395 VL - 3 IS - 3 OP - SN - 0794-4721 UR - http://dx.doi.org/10.4314/apra.v3i3.36395 DB - Crossref ER - TY - CONF TI - Elimination kinetics of tilmicosin following intramammary administration in lactating dairy cows AU - Smith, G.W. AU - Davis, J.L. AU - Baynes, R.E. AU - Riviere, J.E. T2 - Proceedings of the Forty-First Annual Conference, American Association of Bovine Practitioners AB - Tilmicosin is a macrolide antibiotic approved for both the treatment and control of respiratory disease in beef cattle and dairy cattle under 20 months of age. The drug is not approved for use in lactating dairy cattle; however, tilmicosin is sometimes used in an extralabel manner by bovine practitioners for the intramammary treatment of mastitis. This practice results in detectible residues and has resulted in several instances of milk residue violation referred to the Food Animal Residue Avoidance Databank (FARAD). The purpose of this study was to determine the elimination kinetics of tilmicosin in milk following an intramammary infusion so an appropriate proper milk withdrawal interval could be determined. C2 - 2008/9// C3 - Proceedings 41st Annual Convention, American Association of Bovine Practitioners CY - Charlotte, NC DA - 2008/9// PY - 2008/9/25/ DO - 10.21423/aabppro20084428 VL - 41 SP - 230 ER - TY - CONF TI - Ion-Paring Effects on skin absorption of charged nanoparticles AU - Xia, X. AU - Monteiro-Riviere, N.A. AU - Riviere, J.E. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/3// C3 - The Toxicologist CY - Seattle, WA DA - 2008/3// PY - 2008/3/16/ VL - 102 SP - 211 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - CONF TI - Pharmacokinetic modeling and implications of periodicity in arterial extraction of quantum dot nanoparticles AU - Lee, H.A. AU - Imran, M. AU - Mason, S.E. AU - Monteiro-Riviere, N.A. AU - Riviere, J.E. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/3// C3 - The Toxicologist CY - Seattle, WA DA - 2008/3// PY - 2008/3/16/ VL - 102 SP - 287 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - CONF TI - Use of a multi-fiber approach to quantify chemical mixture interactions modulating dermal absorption AU - Imran, M. AU - Baynes, R.E. AU - Xia, X. AU - Riviere, J.E. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/3// C3 - The Toxicologist CY - Seattle, WA DA - 2008/3// PY - 2008/3/16/ VL - 102 SP - 319 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - CONF TI - Comparative effects of surfactants (SLS and LAS) on the dermal absorption of a series of compounds in isolated perfused skin AU - Riviere, James AU - Brooks, J.D. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/3// C3 - The Toxicologist CY - Seattle, WA DA - 2008/3// PY - 2008/3/16/ VL - 102 SP - 320 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - CONF TI - Pharmacokinetics and pharmacokinetic modeling of quantum dots in skin AU - Riviere, J.E. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/3// C3 - The Toxicologist CY - Seattle, WA DA - 2008/3// PY - 2008/3/16/ VL - 102 SP - 406 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - CONF TI - Predicting chemical absorption from complex chemical mixtures AU - Riviere, J.E. T2 - Atlantic Coast Symposium on the Mathematical Sciences in Biology and Biomedicine C2 - 2008/4/26/ C3 - Proceedings of the Atlantic Coast Symposium on the Mathematical Sciences in Biology and Biomedicine CY - North Carolina State University, Raleigh, NC DA - 2008/4/26/ PY - 2008/4/26/ ER - TY - CONF TI - Comparative aspects of topical delivery AU - Riviere, J.E. T2 - 9th Annual Skin Forum C2 - 2008/// C3 - Proceedings of 9th Annual Skin Forum, London CY - London, England, United Kingdom DA - 2008/// PY - 2008/5/26/ SP - 17 ER - TY - CONF TI - Comparative species differences in dermal absorption. Exploiting the similarities and differences for veterinary and human health applications AU - Riviere, J.E. T2 - 35th Annual Meeting and Exposition of the Controlled Release Society C2 - 2008/// C3 - Proceedings of the 35th Annual Meeting Controlled Release Society CY - New York, NY DA - 2008/// PY - 2008/7/12/ ER - TY - CONF TI - Pharmacokinetics of nanomaterials: Is it different from chemicals or drugs? AU - Riviere, J.E. T2 - 2nd International Conference on Nanotoxicology, NANOTOX 2008 C2 - 2008/9// C3 - Proceedings Nano-Tox 2008 CY - Zurich, Switzerland DA - 2008/9// PY - 2008/9/7/ SP - 24/61 ER - TY - CHAP TI - Solvent and vehicle effects on the skin AU - Roberts, M.S. AU - Gierden, A. AU - Riviere, James AU - Monterio-Riviere, N.A. T2 - Dermal Absorption and Toxicity Assessment A2 - Roberts, M.S. A2 - Walters, K.A. PY - 2008/// ET - 2nd SP - 433–447 PB - Informa Healthcare SN - 9780849375910 ER - TY - CONF TI - Disposition of melamine in pigs AU - Baynes, R.E. AU - Smith, G. AU - Mason, S. AU - Barrett, E. AU - Barlow, B.M. AU - Riviere, J.E. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/3// C3 - The Toxicologist CY - Seattle, WA DA - 2008/3// PY - 2008/3/16/ VL - 102 SP - 98 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - CONF TI - The 2007 pet food related toxic nephropathy in dogs and cats. AU - Riviere, J.E. AU - Gupta, R. T2 - 47th Annual Meeting and ToxExpo C2 - 2008/// C3 - The Toxicologist CY - Seattle, WA DA - 2008/// PY - 2008/3/16/ VL - 102 SP - 129 M1 - S1 UR - https://www.toxicology.org/pubs/docs/Tox/2008Tox.pdf ER - TY - JOUR TI - Challenging Hy-Line W36 chicks with KLH in conjunction with retinyl palmitate does not increase antibody response regardless of vitamin A status AU - Livingston, K.A. AU - Perano, T.W. AU - Klasing, K.C. T2 - Poultry Science DA - 2008/// PY - 2008/// VL - 87 IS - Suppl. 1 SP - 83 ER - TY - JOUR TI - Inadequate vitamin A during development increases bursacyte apoptosis in chicks AU - Livingston, K.A. AU - Klasing, K.C. T2 - Poultry Science DA - 2008/// PY - 2008/// VL - 87 IS - Suppl. 1 SP - 52 ER - TY - JOUR TI - Compendium of Veterinary Standard Precautions for Zoonotic Disease Prevention in Veterinary Personnel AU - Elchos, B.L. AU - Scheftel, J.M. AU - Cherry, B. AU - Debess, E.E. AU - Hopkins, Hunter L. AU - Levine, J.F. AU - Williams, C.J. AU - Bell, M.R. AU - Dvorak, G.D. AU - Flora, C.A. AU - Hofmann, J. AU - Pavlin, B.I. AU - Samples, O.M. AU - Snow, J.L. AU - Stinson-Dixon, R.E. T2 - Journal of the American Veterinary Medical Association DA - 2008/8/1/ PY - 2008/8/1/ DO - 10.2460/javma.233.3.415 VL - 233 IS - 3 SP - 415–432 ER - TY - CONF TI - Use of T-RFLP Analysis to Categorize the Microbial Diversity in Native Bivalve Streams in a North Carolina River Basin AU - Bucci, J AU - Caldwell, JM AU - Szempruch, AJ AU - Levine, JF T2 - NC Society For Microbiology C2 - 2008/10// CY - Greensboro, North Carolina DA - 2008/10// PY - 2008/10// ER - TY - MGZN TI - Photo of Wilson Bay AU - Levine, J.F. T2 - NC Alumni Magazine, Talking Trash DA - 2008/// PY - 2008/// M1 - spring ER - TY - RPRT TI - Methods for the preparation and formulation and use of lithium valproate AU - Malkar, N.B. AU - Nelson, D.J. DA - 2008/4/15/ PY - 2008/4/15/ M1 - 7790921 M3 - U.S. Patent SN - 7790921 ER - TY - JOUR TI - Cryotherapy for removal of a premaxillary mass from a chain pickerel using an over-the-counter wart remover AU - Harms, C.A. AU - Christian, L.S. AU - Burrus, O. AU - Hopkins, W.B. AU - Pandiri, A.K.R. AU - Law, J.M. AU - Wolf, K.N. AU - Butler, C.M. AU - Lewbart, G.A. T2 - Exotic DA - 2008/// PY - 2008/// VL - DVM 10 SP - 15–17 ER - TY - CHAP TI - Proventriculitis and proventricular dilatation of broiler chickens AU - Hafner, S. AU - Goodwin, M.A. AU - Guy, J.S. AU - Pantin-Jackwood, M. T2 - Diseases of Poultry A2 - Saif, Y.M. A2 - Fadly, A. A2 - Glisson, J.R. A2 - McDougald, L.R. A2 - Nolan, L.K. A2 - Swayne, D.E. PY - 2008/// ET - 12th SP - 1272-1277 PB - Blackwell/Iowa State University Press ER - TY - CHAP TI - Avian enterovirus infections AU - Guy, J.S. AU - McNulty, M.S. AU - Hayhow, C. T2 - Diseases of Poultry A2 - Saif, Y.M. A2 - Fadley, A. A2 - Glisson, J.R. A2 - McDougald, L.R. A2 - Nolan, L.K. A2 - Swayne, D.E. PY - 2008/// ET - 12th SP - 358–364 PB - Blackwell/Iowa State University Press ER - TY - CHAP TI - Turkey viral hepatitis AU - Guy, J.S. T2 - Diseases of Poultry A2 - Saif, Y.M. A2 - Fadly, A. A2 - Glisson, J.R. A2 - McDougald, L.R. A2 - Nolan, L.K. A2 - Swayne, D.E. PY - 2008/// ET - 12th SP - 428–432 PB - Blackwell/Iowa State University Press ER - TY - CHAP TI - Turkey coronavirus enteritis AU - Guy, J.S. T2 - Diseases of Poultry A2 - Saif, Y.M. A2 - Fadly, A. A2 - Glisson, J.R. A2 - McDougald, L.R. A2 - Nolan, L.K. A2 - Swayne, D.E. PY - 2008/// ET - 12th SP - 330–338 PB - Blackwell/Iowa State University Press ER - TY - CHAP TI - Laryngotracheitis AU - Guy, J.S. AU - Garcia, M. T2 - Diseases of Poultry A2 - Saif, Y.M. A2 - Fadly, A. A2 - Glisson, J.R. A2 - McDougald, L.R. A2 - Nolan, L.K. A2 - Swayne, D.E. PY - 2008/// ET - 12th SP - 137–152 PB - Blackwell/Iowa State University Press ER - TY - CHAP TI - Arbovirus infections AU - Guy, J.S. AU - Malkinson, M. T2 - Diseases of Poultry A2 - Saif, Y.M. A2 - Fadly, A. A2 - Glisson, J.R. A2 - McDougald, L.R. A2 - Nolan, L.K. A2 - Swayne, D.E. PY - 2008/// ET - 12th SP - 416–427 PB - Blackwell/Iowa State University Press ER - TY - CHAP TI - Turkey coronavirus AU - Jackwood, M.W. AU - Guy, J.S. T2 - A laboratory manual for the isolation and identification of avian pathogens A2 - Dufour-Zavala, L. PY - 2008/// ET - 5th SP - 150–152 PB - American Association of Avian Pathologists SN - 9780978916329 ER - TY - CONF TI - Evaluation of a commercial cytology transport fluid for urine sediment preservation AU - Neel, JA AU - Olsson, S T2 - American College of Veterinary Pathologists /American Society for Veterinary Clinical Pathology Annual Meeting C2 - 2008/// C3 - Veterinary Pathology CY - San Antonio, TX DA - 2008/// PY - 2008/11/15/ VL - 45 SP - 791 ER - TY - CHAP TI - Miscellaneous diseases AU - Crespo, R. AU - Shivaprasad, H.L. T2 - Diseases of Poultry A2 - Saif, Y.M. PY - 2008/// ET - 12th SP - 1149-1195 PB - Iowa State Press ER - TY - JOUR TI - Facial Cellulitis Induced in Chickens by Mycoplasma gallisepticum Bacterin and Its Treatment AU - Crespo, Rocio AU - McMillan, Richard T2 - Avian Diseases AB - Ten-to-14-day-old replacement chickens were vaccinated subcutaneously in the neck with a Mycoplasma gallisepticum bacterin. A few days later, 40% of the flock developed cellulitis of the head. Birds were submitted for diagnostic investigation between 4 and 6 wk of age. Histology revealed lesions in the subcutis consistent with oil-emulsified vaccine/bacterin reaction. No significant bacteria were isolated from the lesions. Birds were treated with 1 mg of methyl prednisol acetate intramuscularly. Improvement was noted as early as 12 hr posttreatment. One week after treatment only 1%-2% of the flock still had facial cellulitis. DA - 2008/12// PY - 2008/12// DO - 10.1637/8342-050108-case.1 VL - 52 IS - 4 SP - 698-701 J2 - Avian Diseases LA - en OP - SN - 0005-2086 1938-4351 UR - http://dx.doi.org/10.1637/8342-050108-case.1 DB - Crossref KW - bacterin KW - cellulitis KW - chicken KW - methyl prednisol acetate ER - TY - JOUR TI - T1250 Luminal Uptake of L-Arginine By C. Parvum Infected Porcine Ileum Is Promoted By Epithelial Induction of Arginase II and Stimulates Prostaglandin-Dependent Secretory Diarrhea AU - Gookin, Jody L. AU - Foster, Derek M. AU - Stauffer, Stephen H. AU - Stone (Coccaro), Maria R. T2 - Gastroenterology DA - 2008/4// PY - 2008/4// DO - 10.1016/S0016-5085(08)62407-5 VL - 134 IS - 4 SP - A-515-A-516 J2 - Gastroenterology LA - en OP - SN - 0016-5085 UR - http://dx.doi.org/10.1016/S0016-5085(08)62407-5 DB - Crossref ER - TY - CHAP TI - Program for Monitoring Emerging Diseases ProMED-mail AU - Cowen, P. T2 - Achieving Sustainable Global Capacity for Surveillance and Response to Emerging Diseases of Zoonotic Origin A2 - Beatty, A. A2 - Scott, K. A2 - Tsai, P. PY - 2008/// SP - 57–59 PB - Institute of Medicine, National Academies Press ER - TY - JOUR TI - Coxiella-Like Infection in Psittacines and a Toucan AU - Shivaprasad, H. L. AU - Cadenas, M. B. AU - Diab, S. S. AU - Nordhausen, R. AU - Bradway, D. AU - Crespo, R. AU - Breitschwerdt, E. B. T2 - Avian Diseases AB - Seven psittacine birds and a toucan (Ramphastos toco) were diagnosed as infected with Coxiella-like bacteria, based on polymerase chain reaction and bacterial 16S rRNA gene sequence obtained from each bird's liver tissue. Most of the birds exhibited lethargy and weakness for several days prior to death. Gross lesions included mild to moderate emaciation and severely enlarged and mottled pale livers and spleens. Microscopically, there was multifocal necrosis of hepatocytes with infiltration of a mixed population of inflammatory cells, including lymphocytes, heterophils, plasma cells, and macrophages randomly scattered throughout in most birds. In several birds within the macrophages there were vacuoles containing basophilic small cocco-bacilli organisms measuring about 0.5-1 microm. The spleens had increased numbers of mononuclear phagocytic system cells, some of which had vacuoles that contained similar organisms, as observed in the liver. There was inflammation in the epicardium and endocardium, interstitium of the lungs, kidney, adrenal and thyroid glands, lamina propria of the intestine, and in occasional birds in the brain, bursa of Fabricius, and bone marrow associated with similar organisms in the macrophages. Transmission electron microscopy of the liver and lungs in most birds and in the thyroid glands of one bird revealed pleomorphic round to elongated bacteria measuring about 0.45 microm in diameter and more than 1.0 microm in length. Most of these organisms contained a peripheral zone of loosely arranged electron dense material that was located immediately beneath a trilaminar membrane. Occasional organisms contained nucleoids. This is the first documentation of disease presumptively associated with Coxiella-like bacteria in birds. DA - 2008/9// PY - 2008/9// DO - 10.1637/8192-120707-reg VL - 52 IS - 3 SP - 426-432 J2 - Avian Diseases LA - en OP - SN - 0005-2086 1938-4351 UR - http://dx.doi.org/10.1637/8192-120707-reg DB - Crossref KW - avian KW - psittacines KW - toucan KW - hepatitis KW - splenitis KW - Coxiella ER - TY - JOUR TI - Interaction of Ionophore and Vitamin E in Knockdown Syndrome of Turkeys AU - Crespo, Rocio AU - Shivaprasad, H. L. AU - Sommer, Franz AU - Puschner, Birgit T2 - Journal of Veterinary Diagnostic Investigation AB - Monensin and vitamin E concentrations, as well as histopathology of skeletal muscles and myocardium, were evaluated in broad-breasted white turkeys kept in commercial facilities. Turkeys with knockdown syndrome had myopathy of skeletal muscles, but no lesions in the myocardium. Generally, concentration of monensin in serum was highest in turkeys diagnosed with knockdown syndrome given more than 90 mg/kg of monensin in the diet, followed by turkeys diagnosed with knockdown syndrome given <90 mg/kg of monensin in the diet, healthy turkeys fed a diet that contained <90 mg/kg of monensin, and finally healthy turkeys fed a diet free of monensin (not detectable). However, the concentration of monensin was highly variable within each group, and the median was lower than the average. Vitamin E concentrations in the livers varied from low-normal to below normal and were statistically higher in healthy turkeys fed a diet free of monensin than in the livers of birds from the 3 groups exposed to monensin. This suggests that the concentration of monensin in serum positively correlates to the severity of clinical signs and pathology and to the amount of monensin in the feed. Although the methodology developed to detect serum monensin concentrations is beneficial and accurate for case investigations, it is recommended that several samples from each flock be evaluated because of variation within a flock. The current study also suggests that monensin in the feed could induce lower concentrations of vitamin E in the liver of turkeys and can predispose the turkeys to knockdown syndrome. DA - 2008/7// PY - 2008/7// DO - 10.1177/104063870802000409 VL - 20 IS - 4 SP - 472-476 J2 - J VET Diagn Invest LA - en OP - SN - 1040-6387 1943-4936 UR - http://dx.doi.org/10.1177/104063870802000409 DB - Crossref KW - Ionophore KW - knockdown syndrome KW - monensin KW - turkeys KW - vitamin E ER - TY - JOUR TI - Bilateral Malacia Associated with Sodium Poisoning in Turkey Poults AU - Crespo, Rocio AU - Subbiah, Murugan AU - Corsiglia, Charles AU - Bickford, Arthur AU - Puschner, Birgit T2 - Avian Diseases AB - Improper cleaning of the water storage tank resulted in a toxic concentration of sodium in drinking water in a commercial turkey flock. Within the first week after placement 40% of the birds in the flock died. Clinically, poults were depressed and weak, huddled together, and reluctant to walk. At necropsy the birds had crops and gizzards filled with rice hulls, moderately swollen livers, distended gall bladders, and congested lungs. Neither ascites nor round heart was observed. The major microscopic lesion was multifocal symmetrical malacia of brain and spinal cord. Laboratory results revealed a high concentration of sodium in water (2340 mg/liter). The concentration of sodium in brain and liver ranged from 1870 to 2680 (mean = 2185; SD = 321.5) mg/liter wet weight and from 1810 to 2360 (mean = 2191.67; SD = 193.2) mg/liter wet weight, respectively, whereas the normal expected sodium concentration in the brain and liver tissues from young turkeys (<7 days old) that were submitted for other causes averaged 1233 and 983 mg/liter wet weight, respectively. Based on the histological and toxicological results, a diagnosis of salt poisoning was made. This case investigation demonstrated that sodium analysis of brain and liver are diagnostically useful when confirming sodium poisoning in young turkeys. DA - 2008/3// PY - 2008/3// DO - 10.1637/8025-053107-case VL - 52 IS - 1 SP - 179-182 J2 - Avian Diseases LA - en OP - SN - 0005-2086 1938-4351 UR - http://dx.doi.org/10.1637/8025-053107-case DB - Crossref KW - Malacia KW - poisoning KW - poult KW - toxicity KW - turkey KW - salt KW - sodium ER - TY - CHAP TI - Isolation and Propagation of Coronaviruses in Embryonated Eggs AU - Guy, James S. T2 - Methods in Molecular Biology A2 - Cavanagh, D. AB - The embryonated egg is a complex structure comprising an embryo and its supporting membranes (chorioallantoic, amniotic, yolk). The developing embryo and its membranes provide the diversity of cell types that are needed for successful replication of a wide variety of different viruses. Within the family Coronaviridae, the embryonated egg has been used as a host system primarily for two group 3 coronaviruses, infectious bronchitis virus (IBV) and turkey coronavirus (TCoV), but it also has been shown to be suitable for pheasant coronavirus. IBV replicates well in the embryonated chicken egg, regardless of the inoculation route; however, the allantoic route is favored as the virus replicates extensively in chorioallantoic membrane and high titers are found in allantoic fluid. TCoV replicates only in embryo tissues, within epithelium of the intestines and bursa of Fabricius; thus amniotic inoculation is required for isolation and propagation of this virus. Embryonated eggs also provide a potential host system for studies aimed at identifying other, novel coronavirus species. PY - 2008/// DO - 10.1007/978-1-59745-181-9_10 VL - 454 SP - 109-117 PB - Humana Press SN - 9781588298676 9781597451819 UR - http://dx.doi.org/10.1007/978-1-59745-181-9_10 ER - TY - JOUR TI - Multilocus Sequence Typing Analysis of Clostridium perfringens Isolates from Necrotic Enteritis Outbreaks in Broiler Chicken Populations AU - Chalmers, G. AU - Bruce, H. L. AU - Hunter, D. B. AU - Parreira, V. R. AU - Kulkarni, R. R. AU - Jiang, Y.-F. AU - Prescott, J. F. AU - Boerlin, P. T2 - Journal of Clinical Microbiology AB - Clostridium perfringens is an important pathogen of animals and humans and is the causative agent of necrotic enteritis (NE) in poultry. This study focuses on the typing of intestinal C. perfringens isolates (n = 61) from outbreaks of NE collected from several areas of Southern Ontario, using a recently developed multilocus sequence typing (MLST) technique. For comparison, C. perfringens isolates from healthy birds were also obtained and typed. An additional locus, the pfoS locus, was included in our analysis, in an attempt to increase the discriminatory ability of the method previously published. Birds were collected from two major poultry processors in Canada, and isolates from processor 2 formed a distinct MLST cluster. Isolates from healthy birds also collected from the outbreak flocks clustered together with isolates from the birds with NE. Although isolates from eight outbreaks clustered together, MLST types were also occasionally different between outbreaks. Strong linkage disequilibrium was observed between loci, suggesting a clonal C. perfringens population structure. Detection assays for toxin genes cpb2 (beta-2 toxin), tpeL, and the newly described netB (NetB toxin) were also performed. netB was almost always found in outbreak isolates, whereas cpb2 was found exclusively in healthy bird isolates. The toxin gene tpeL, which has not been previously identified in C. perfringens type A strains, was also found, but only in the presence of netB. Resistance to bacitracin was found in 34% of isolates from antimicrobial agent-free birds and in 100% of isolates from conventionally raised birds. DA - 2008/10/22/ PY - 2008/10/22/ DO - 10.1128/jcm.01548-08 VL - 46 IS - 12 SP - 3957-3964 J2 - Journal of Clinical Microbiology LA - en OP - SN - 0095-1137 UR - http://dx.doi.org/10.1128/jcm.01548-08 DB - Crossref ER - TY - JOUR TI - Oral immunization of broiler chickens against necrotic enteritis with an attenuated Salmonella vaccine vector expressing Clostridium perfringens antigens AU - Kulkarni, R.R. AU - Parreira, V.R. AU - Sharif, S. AU - Prescott, J.F. T2 - Vaccine AB - Necrotic enteritis (NE) in broiler chickens is caused by Clostridium perfringens but currently no effective vaccine is available. Our previous study showed that certain C. perfringens secreted proteins when administered intramuscularly protected chickens against experimental infection. In the current study, genes encoding three C. perfringens proteins: fructose-biphosphate-aldolase (FBA), pyruvate:ferredoxin-oxidoreductase (PFOR) and hypothetical protein (HP), were cloned into an avirulent Salmonella enterica sv. typhimurium vaccine vector. Broiler chickens immunized orally with recombinant Salmonella expressing FBA or HP proteins were significantly protected against NE challenge. Immunized birds developed serum and mucosal antibodies to both clostridial and Salmonella antigens. This study showed the oral immunizing ability of two C. perfringens antigens against NE in broiler chickens through an attenuated Salmonella vaccine vector. DA - 2008/8// PY - 2008/8// DO - 10.1016/j.vaccine.2008.05.079 VL - 26 IS - 33 SP - 4194-4203 J2 - Vaccine LA - en OP - SN - 0264-410X UR - http://dx.doi.org/10.1016/j.vaccine.2008.05.079 DB - Crossref KW - Clostrdium perfringens KW - vaccine KW - necrotic enteritis KW - chickens ER - TY - JOUR TI - 10 INOCULATION OF CULTURE-NEGATIVE PORCINE SEMEN WITH NOVEL BIOFILM-FORMING BACTERIA AU - Clark, S. AU - Ness, A. AU - Baldrighi, J. AU - Borst, L. AU - Maddox, C. AU - Payne, B. T2 - Reproduction, Fertility and Development AB - With the growing number of boar studs having semen analysis performed by reproductive specialists, a growing number of diagnostic challenges are encountered. Semen analysis classically involves evaluation of sperm cell motility, morphology, and concentration; however, culture of the extended semen sample for bacterial contamination has become routine. Two isolates, Achromobacter xylosoxidans and Ralstonia pickettii, have recently been identified in the water distillation system of a boar stud facility that uses this water to extend the raw semen in various semen extenders. Insemination of sows with contaminated semen has resulted in severe pyometras diagnosed on necropsy. The effect of these bacteria on sperm motility has not been examined in a controlled setting. The objective of this study was to determine the effects of A. xylosoxidans (AX) and R. pickettii (RP) on pH and motility in culture-negative semen samples over a 7-day period at 16�C. Banked clinical isolates of AX and RP were plated on Columbia blood agar and incubated for 48 h at 37�C. For each isolate, a single colony was selected and transferred to 10 mL of Luria broth. The broth was then incubated for 24 h at 37�C. Optical density measurements were performed at 24 and 48 h of growth, followed by quantification of bacteria by plate counts of serially diluted broth cultures (colony forming units). At 24 h, AX and RP reached levels of 1 � 108 and 1 � 107 [colony-forming units (cfu) mL–1], respectively. Concentration of bacteria in clinical infection was determined to be approximately 1 � 104 and 102 for AX and RP, respectively. In order to attain concentrations similar to those in clinical infection, dilution of the bacteria was necessary. Centrifugation of broth culture at 4000 rpm for 5 min was performed and the bacterial pellet was re-suspended in culture-negative semen in Modena (SGI, LTD, Cambridge, IA, USA) extender to concentrations mimicking those in clinical infection. The samples were then incubated at 16�C and rotated once daily. Motility and morphology, viewed using computer-assisted sperm analysis (CASA: Spermvision; Minitube of America, Verona, WI, USA), and pH (Accumet AB15, Fisher Scientific, Hanover Park, IL, USA) were measured daily for each sample at 25�C. Data from 4 replicates were used in the analysis. For motility, ANOVA revealed no significant differences (P < 0.05) between the control and inoculated samples. A PROX MIXED analysis (SAS, SAS Institute, Inc., Cary, NC, USA) revealed no treatment-by-time interaction with sperm motility after inoculation. For sample pH, statistically significant differences (P < 0.05) were noted between all of the samples, primarily contributed by a treatment-by-time effect. The pH of the control sample became more basic over the 7-day period (from 6.94 to 7.32). This phenomenon was also observed in all of samples; however, semen inoculated with AX appeared to remain closer to neutral pH than did the RP samples. Although statistically significant differences were noted in pH, the addition of biofilm bacteria did not negatively affect the motility of extended porcine semen during this time period. Further experiments need to be performed in relation to different concentrations, time period of bacterial growth, and determination of final cfu mL–1. DA - 2008/// PY - 2008/// DO - 10.1071/rdv20n1ab10 VL - 20 IS - 1 SP - 85 J2 - Reprod. Fertil. Dev. LA - en OP - SN - 1031-3613 UR - http://dx.doi.org/10.1071/rdv20n1ab10 DB - Crossref ER - TY - JOUR TI - Distribution of viral antigen gp85 and provirus in various tissues from commercial meat-type and experimental White Leghorn Line 0 chickens with different subgroup J avian leukosis virus infection profiles AU - Pandiri, A. R. AU - Gimeno, I. M. AU - Reed, W. M. AU - Lee, L. F. AU - Silva, R. F. AU - Fadly, A. M. T2 - Avian Pathology AB - Immunohistochemistry and polymerase chain reaction (PCR) were used to test for the presence of avian leukosis virus (ALV) J viral antigen gp85 and proviral DNA, respectively, in various tissues (adrenal gland, bone marrow, gonad, heart, kidney, liver, lung, pancreas, proventriculus, sciatic nerve, spleen, and thymus). Tissues were collected from 32-week-old commercial meat-type and Avian Disease and Oncology Laboratory experimental White Leghorn Line 0 chickens with the following different infection profiles: tV + A−, included in ovo-tolerized viraemic chickens with no neutralizing antibodies (NAbs) on any sampling; ntV + A−, included chickens that were viraemic and NAb-negative at the time of termination at 32 weeks post hatch, but had NAbs on up to two occasions; V+ A+, included chickens that were viraemic and NAb-positive at the time of termination at 32 weeks post hatch, and had NAbs on more than two occasions; V − A+, included chickens that were negative for viraemia and NAb-positive at the time of termination at 32 weeks post hatch, and had antibody on more than two occasions; V − A−, included chickens that were never exposed to ALV J virus. There was a direct correlation between viraemia and tissue distribution of gp85, regardless of the NAb status and strain of chickens, as expression of ALV J gp85 was noted in only viraemic chickens (tV + A−, ntV + A−, V+ A+), but not in non-viraemic seroconverted chickens (V − A+). Of the four oligonucleotide primers pairs used in PCR to identify ALV J provirus, only one primer set termed H5/H7 was useful in demonstrating ALV J proviral DNA in the majority of the tissues tested from non-viraemic, antibody-positive chickens (V − A+). The results suggest that PCR using primer pair H5/H7 is more sensitive than immunohistochemistry in identifying ALV J in chickens that have been exposed to virus, but are not actively viraemic. DA - 2008/1/16/ PY - 2008/1/16/ DO - 10.1080/03079450701774843 VL - 37 IS - 1 SP - 7-13 SN - 0307-9457 1465-3338 UR - http://dx.doi.org/10.1080/03079450701774843 ER - TY - JOUR TI - Posterior paresis resulting from a vertebral body fracture in a bull AU - Foster, D.M. AU - Smith, G.W. AU - Pandiri, A.R. AU - Blond, L.R. T2 - The Bovine Practitioner AB - A 2.5-year-old, mixed-breed rodeo bull was examined with a four-month history of ataxia and weakness in the pelvic limbs. The neurologic signs were noted while the bull was on pasture breeding cows. Spinal radiographs revealed a compression fracture of the 13th thoracic vertebrae, and myelography showed impingement of the spinal cord due to the fracture. Necropsy examination confirmed the diagnosis of a compression fracture of T13 with secondary damage to the spinal cord. Vertebral fractures are uncommon in adult cattle, yet this case appears to be unrelated to any previously published causes. Furthermore, myelography can be an effective diagnostic test in the evaluation of spinal cord disease in adult cattle. DA - 2008/// PY - 2008/// DO - 10.21423/bovine-vol42no1p41-44 VL - 42 IS - 1 SP - 41–44 ER - TY - JOUR TI - The effect of heterogeneous infectious period and contagiousness on the dynamics of Salmonella transmission in dairy cattle T2 - Epidemiology and Infection AB - SUMMARY The objective of this study was to address the impact of heterogeneity of infectious period and contagiousness on Salmonella transmission dynamics in dairy cattle populations. We developed three deterministic SIR-type models with two basic infected stages (clinically and subclinically infected). In addition, model 2 included long-term shedders, which were defined as individuals with low contagiousness but long infectious period, and model 3 included super-shedders (individuals with high contagiousness and long infectious period). The simulated dynamics, basic reproduction number ( R 0 ) and critical vaccination threshold were studied. Clinically infected individuals were the main force of infection transmission for models 1 and 2. Long-term shedders had a small impact on the transmission of the infection and on the estimated vaccination thresholds. The presence of super-shedders increases R 0 and decreases the effectiveness of population-wise strategies to reduce infection, making necessary the application of strategies that target this specific group. DA - 2008/// PY - 2008/// DO - 10.1017/S0950268807000209 VL - 136 IS - 11 SP - 1496-1510 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-54049121463&partnerID=MN8TOARS ER - TY - JOUR TI - The risk and control of Salmonella outbreaks in calf-raising operations: A mathematical modeling approach T2 - Veterinary Research AB - Salmonellosis in calves has economic and welfare implications, and serves as a potential source of human infections. Our objectives were to assess the risk of Salmonella spread following its introduction into a herd of pre-weaned calves and to evaluate the efficacy of control strategies to prevent and control outbreaks. To meet these objectives, we developed a model of Salmonella transmission within a pre-weaned group of calves based on a well documented outbreak of salmonellosis in a calf-raising operation and other literature. Intervention scenarios were evaluated in both deterministic and stochastic versions of the model. While the basic reproduction number (R0) was estimated to be 2.4, simulation analysis showed that more than 60% of the invasions failed after the introduction of a single index case. With repeated introduction of index cases, the probability of Salmonella spread was close to 1, and the tested control strategies were insufficient to prevent transmission within the group. The most effective strategies to control ongoing outbreaks were to completely close the rearing operation to incoming calves, to increase the proportion of admitted calves that were immunized (>75%), and to assign personnel and equipment to groups of calves. DA - 2008/// PY - 2008/// DO - 10.1051/vetres:2008038 VL - 39 IS - 6 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-55549086869&partnerID=MN8TOARS ER - TY - JOUR TI - Improved feed protein fractionation schemes for formulating rations with the cornell net carbohydrate and protein system AU - Lanzas, C. AU - Broderick, G.A. AU - Fox, D.G. T2 - Journal of Dairy Science AB - Adequate predictions of rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) supplies are necessary to optimize performance while minimizing losses of excess nitrogen (N). The objectives of this study were to evaluate the original Cornell Net Carbohydrate Protein System (CNCPS) protein fractionation scheme and to develop and evaluate alternatives designed to improve its adequacy in predicting RDP and RUP. The CNCPS version 5 fractionates CP into 5 fractions based on solubility in protein precipitant agents, buffers, and detergent solutions: A represents the soluble nonprotein N, B1 is the soluble true protein, B2 represents protein with intermediate rates of degradation, B3 is the CP insoluble in neutral detergent solution but soluble in acid detergent solution, and C is the unavailable N. Model predictions were evaluated with studies that measured N flow data at the omasum. The N fractionation scheme in version 5 of the CNCPS explained 78% of the variation in RDP with a root mean square prediction error (RMSPE) of 275 g/d, and 51% of the RUP variation with RMSPE of 248 g/d. Neutral detergent insoluble CP flows were overpredicted with a mean bias of 128 g/d (40% of the observed mean). The greatest improvements in the accuracy of RDP and RUP predictions were obtained with the following 2 alternative schemes. Alternative 1 used the inhibitory in vitro system to measure the fractional rate of degradation for the insoluble protein fraction in which A = nonprotein N, B1 = true soluble protein, B2 = insoluble protein, C = unavailable protein (RDP: R(2) = 0.84 and RMSPE = 167 g/d; RUP: R(2) = 0.61 and RMSPE = 209 g/d), whereas alternative 2 redefined A and B1 fractions as the non-amino-N and amino-N in the soluble fraction respectively (RDP: R(2) = 0.79 with RMSPE = 195 g/d and RUP: R(2) = 0.54 with RMSPE = 225 g/d). We concluded that implementing alternative 1 or 2 will improve the accuracy of predicting RDP and RUP within the CNCPS framework. DA - 2008/// PY - 2008/// DO - 10.3168/jds.2008-1440 VL - 91 IS - 12 SP - 4881-4891 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-57749117028&partnerID=MN8TOARS KW - feed protein fractionation KW - protein supply KW - nutritional model ER - TY - JOUR TI - Clinical, clinicopathological and histological changes observed in 14 cats treated with glucocorticoids. AU - Lowe, AD AU - Campbell, KL AU - Barger, A AU - Schaeffer, DJ AU - Borst, L T2 - The Veterinary record AB - Fourteen cats were given immunosuppressive doses of either prednisolone (4·4 mg/kg/day) or dexamethasone (0·55 mg/kg/day) for 56 days. Complete blood counts, serum biochemistry profiles and urinalyses were performed on days 0 and 56, and liver biopsies were taken laparoscopically on day 56, because of evidence of hepatic disease on the serum biochemistry profiles. There were significant increases in the cats' mean white blood cell counts, neutrophil counts and monocyte counts, and significant decreases in their mean lymphocyte counts and eosinophil counts. There were consistent increases in the serum concentrations of albumin, glucose, triglycerides and cholesterol. Glycogen deposition, consistent with a steroid hepatopathy, was present to varying degrees in all the liver biopsies. One of the cats developed adverse clinical signs including anorexia, icterus, pruritus and medial curling of the pinnae, some of which were suspected to be related to the glucocorticoid therapy. DA - 2008/6// PY - 2008/6// DO - 10.1136/vr.162.24.777 VL - 6 UR - http://europepmc.org/abstract/med/18552328 ER - TY - JOUR TI - Antimicrobial Resistance and Virulence of Enterococcus faecalis Isolated from Retail Food AU - McGowan-Spicer, Lori L. AU - Fedorka Cray, Paula J. AU - Frye, Jonathan G. AU - Meinersmann, Richard J. AU - Barrett, John B. AU - Jackson, Charlene R. T2 - Journal of Food Protection AB - Although enterococci are considered opportunistic nosocomial pathogens, their contribution to foodborne illnesses via dissemination through retail food remains undefined. In this study, prevalence and association of antimicrobial resistance and virulence factors of 80 Enterococcus faecalis isolates from retail food items were investigated. The highest rates of resistance were observed for lincomycin (73 of 80 isolates, 91%) and bacitracin (57 of 80 isolates, 71%), and lower rates of resistance (< or =40%) were found for chloramphenicol, ciprofloxacin, erythromycin, flavomycin, gentamicin, kanamycin, nitrofurantoin, penicillin, and tylosin. Overall resistance to antimicrobials was low for most isolates tested. Of the virulence factors tested, the majority of isolates were positive for ccf (78 of 80 isolates, 98%), efaAfs (77 of 80, 96%), and cpd (74 of 80, 93%). Isolates also commonly contained cob (72 of 80 isolates, 90%) and gelE (68 of 80, 85%). Very few isolates contained cylMBA (12 of 80 isolates [15%] for cylM and 9 of 80 isolates [11%] for both cylB and cylA) and efaAfm (2 of 80 isolates, 3%). Positive statistical associations (significance level of 0.05) were found between agg and tetracycline resistance, cylM and erythromycin resistance, and gelE and efaAfs and lincomycin resistance. The presence of the cylB and cylA alleles also was positively correlated with bacitracin and tetracycline resistance. Negative correlations were observed between many of the virulence attributes and resistance to ciprofloxacin, erythromycin, flavomycin, gentamicin, kanamycin, and tylosin. These data suggest that both positive and negative associations exist between antimicrobial resistance genes and virulence factors in E. faecalis isolates from foods commonly purchased from grocery stores. DA - 2008/4// PY - 2008/4// DO - 10.4315/0362-028x-71.4.760 VL - 71 IS - 4 SP - 760-769 SN - 0362-028X UR - http://dx.doi.org/10.4315/0362-028x-71.4.760 ER - TY - JOUR TI - Detection of Campylobacter from Broiler Carcass Rinse Samples Utilizing the Tecra Visual Immunoassay (via) AU - Bailey, J.S. AU - Fedorka Cray, P. AU - Richardson, L.J. AU - Cox, N.A. AU - Cox, J.M. T2 - Journal of Rapid Methods & Automation in Microbiology AB - ABSTRACT Poultry meat is considered to be a major vector of transmission of Campylobacter, either directly, through consumption of poorly prepared product or cross‐contamination, or indirectly, through introduction of the bacterium into the food production environment. Efficient detection of Campylobacter is intrinsic to the management of the pathogen during poultry production. The TECRA Campylobacter Visual Immunoassay (CAMVIA) protocol, enrichment in a proprietary TECRA Campylobacter enrichment broth followed by an ELISA, was compared with a conventional cultural method, with enrichment in Bolton medium (containing lysed horse blood), followed by plating onto Campy‐cefex agar. Of the 398 broiler carcass rinses tested from 19 commercial processing plants, a total of 350 carcasses (88%) were found to be positive for Campylobacter by at least one method. The TECRA CAMVIA yielded 317 Campylobacter ‐positive results, four of which were false positive and 22 of which were false negative, while conventional enrichment and plating detected the bacterium in only 280 samples with 48 false negatives. These data demonstrate that the TECRA CAMVIA has a statistically higher sensitivity and specificity than a conventional culture procedure using Bolton broth as the enrichment. PRACTICAL APPLICATIONS Standard Campylobacter methodology procedures currently take 4–5 days for detection of Campylobacter spp. in a sample when using an enrichment step. The TECRA Campylobacter Visual Immunoassay (VIA) reduces the amount of time it takes to determine whether Campylobacter spp. are present on poultry carcasses that have been rinsed. The procedure can effectively determine the presence of Campylobacter spp. in 2 days and has a statistically higher sensitivity and specificity than a conventional culture procedure using Bolton broth as the enrichment. DA - 2008/12// PY - 2008/12// DO - 10.1111/j.1745-4581.2008.00142.x VL - 16 IS - 4 SP - 374-380 SN - 1060-3999 1745-4581 UR - http://dx.doi.org/10.1111/j.1745-4581.2008.00142.x ER - TY - JOUR TI - Antimicrobial Resistance of Listeria monocytogenes Isolated from a Poultry Further Processing Plant AU - Lyon, Steven A. AU - Berrang, Mark E. AU - Fedorka-Cray, Paula J. AU - Fletcher, Daniel L. AU - Meinersmann, Richard J. T2 - Foodborne Pathogens and Disease AB - The aim of this study was to compare antimicrobial resistance profiles of distinct types of Listeria monocytogenes isolated from a commercial poultry cooking plant. One hundred fifty-seven L. monocytogenes isolates representing 14 different actA types were tested for susceptibility to 19 antimicrobial drugs using the broth microdilution method. All isolates were susceptible to antimicrobials except for ceftriaxone (153 isolates [97%] intermediate or resistant), oxacillin with 2% NaCl (142 isolates [90%] resistant), ciprofloxacin (59 isolates [37%] intermediate or resistant), tetracycline (5 isolates [3%] resistant), clindamycin (43 isolates [27%] intermediate), linezolid (3 isolates [2%] intermediate), and trimethoprim/sulfamethoxazole (1 isolate [<1%] intermediate). Prevalence of antimicrobial resistance was low among all actA types. There was a low amount of diversity of resistotypes, which were defined in this study as subdivisions of actA types according to antimicrobial resistance profiles of the isolates. The five tetracycline-resistant isolates represented all the members of one actA type in lineage II. This study showed that antimicrobial resistance is not highly prevalent in L. monocytogenes from a poultry further processing environment. Types of L. monocytogenes as distinguished by actA sequencing do not predict antimicrobial resistance except possibly for tetracycline resistance. L. monocytogenes types that persist in a poultry cook plant are not related to antimicrobial resistance, and excluding tetracycline resistance, antimicrobial resistance does not seem to differ according to actA type or lineage. DA - 2008/6// PY - 2008/6// DO - 10.1089/fpd.2007.0070 VL - 5 IS - 3 SP - 253-259 SN - 1535-3141 1556-7125 UR - http://dx.doi.org/10.1089/fpd.2007.0070 ER - TY - JOUR TI - Characterization of Shiga Toxin–Producing Escherichia coli Strains Isolated from Swine Feces AU - Fratamico, Pina M. AU - Bhagwat, Arvind A. AU - Injaian, Lisa AU - Fedorka-Cray, Paula J. T2 - Foodborne Pathogens and Disease AB - The virulence gene and antibiotic resistance profiles of Shiga toxin-producing Escherichia coli (STEC) strains belonging to 58 different O:H serotypes (219 strains) isolated from swine feces were determined. Of the 219 isolates, 29 (13%) carried the stx(1) gene, 14 (6%) stx(2), 176 (80%) stx(2e), 46 (21%) estIa, 14 (6.4%) estIb, 10 (4.6%) fedA, 94 (42.9%) astA, 25 (11.4%) hly(933), and one (0.46%) cdt-III. None of the strains possessed the elt, bfp, faeG, fanA, fasA, fimF(41a), cnf-1, cnf-2, eae, cdt-I, or cdt-IV genes. The strains were also tested for antibiotic susceptibility using 16 antibiotics. The STEC isolates displayed resistance most often to tetracycline (95.4%), sulfamethoxazole (53.4%), kanamycin (38.4%), streptomycin (34.7%), and chloramphenicol (22.4%). An E. coli serotype O20:H42 strain, which was positive for stx(2e) and astA, was resistant to all of the antibiotics tested except for amikacin. In addition, 52 of the swine isolates, representing 16 serogroups and 30 different serotypes, were examined for their ability to withstand acid challenge by three types of acid resistance (AR) pathways, AR1 (rpoS dependent), AR2 (glutamate dependent), and AR3 (arginine dependent). None of the strains was defective in the AR1 resistance pathway, while one strain was defective in the AR2 pathway under aerobic growth conditions but fully functional under anaerobic growth conditions. Among the three AR pathways, the AR3 pathway offered the least protection, and 8 out of 52 strains were defective in this pathway. The strain that was defective in AR2 was fully functional in the AR3 pathway. Since AR plays a vital role in the survival and virulence of these strains, differences among the isolates to induce AR pathways may play a significant role in determining their infective dose. This study demonstrates that swine STEC comprise a heterogeneous group of organisms, and the possession of different combinations of E. coli virulence genes indicate that some swine STEC can potentially cause human illness. DA - 2008/12// PY - 2008/12// DO - 10.1089/fpd.2008.0147 VL - 5 IS - 6 SP - 827-838 J2 - Foodborne Pathogens and Disease LA - en OP - SN - 1535-3141 1556-7125 UR - http://dx.doi.org/10.1089/fpd.2008.0147 DB - Crossref ER - TY - JOUR TI - Analysis of Salmonella enterica with Reduced Susceptibility to the Third-Generation Cephalosporin Ceftriaxone Isolated from U.S. Cattle During 2000–2004 AU - Frye, Jonathan G. AU - Fedorka-Cray, Paula J. AU - Jackson, Charlene R. AU - Rose, Markus T2 - Microbial Drug Resistance AB - Over the past decade enteric bacteria in Europe, Africa, and Asia have become increasingly resistant to cephalosporin antimicrobial agents. This is largely due to the spread of genes encoding extended-spectrum beta-lactamase (ESBL) enzymes that can inactivate many cephalosporins. Recently, these resistance mechanisms have been identified in Salmonella isolated from humans in the United States. Due to the potential for transmission of resistant bacteria to humans via food animals, Salmonella animal isolates were monitored for ESBL production. During 2000-2004, Salmonella cattle slaughter isolates (n = 3,984) were tested, and 97 (2.4%) of these were found to have decreased susceptibility (minimum inhibitory concentration [MIC] >32 microg/ml) to the third-generation cephalosporin ceftriaxone. The majority of these were serotypes Newport (58) and Agona (14), some of which were genetically indistinguishable by pulsed field gel electrophoresis (PFGE) analysis. None of the isolates had an ESBL phenotype; all were susceptible to the fourth-generation cephalosporins cefepime and cefquinome. PCR and sequence analysis for resistance genes detected the bla(CMY-2) gene in 93 isolates and the bla(TEM-1) gene in 12 isolates; however, neither gene encodes an ESBL. These data indicate that bovine Salmonella isolates from the United States with decreased susceptibility or resistance to ceftriaxone do not exhibit an ESBL phenotype and most contain the bla(CMY-2) gene. DA - 2008/12// PY - 2008/12// DO - 10.1089/mdr.2008.0844 VL - 14 IS - 4 SP - 251-258 J2 - Microbial Drug Resistance LA - en OP - SN - 1076-6294 1931-8448 UR - http://dx.doi.org/10.1089/mdr.2008.0844 DB - Crossref ER - TY - JOUR TI - Fecal Shedding of Foodborne Pathogens by Florida-Born Heifers and Steers in U.S. Beef Production Segments AU - Riley, D. G. AU - Loneragan, G. H. AU - Phillips, W. A. AU - Gray, J. T. AU - Fedorka Cray, P. J. T2 - Journal of Food Protection AB - The objective in this study was to assess breed effects in fecal prevalence of Escherichia coli O157:H7 in heifers on a development program in Florida and in their steer half siblings in stocker and feedlot phases in Oklahoma. A secondary objective was to characterize fecal shedding of Campylobacter and Salmonella in subsets of the same samples. After weaning, heifers (n = 501; purebreds and F1 crosses of Angus, Brahman, and Romosinuano) were preconditioned and placed in a local development program. Steers (n = 481) were transported to Oklahoma, where they grazed wheat for 6 months and then were placed in feedlot pens. Fecal samples were obtained at least every 28 days for 12 months on most animals. None of the 10,982 samples tested positive for E. coli O157:H7. Overall fecal prevalences of Campylobacter and Salmonella in heifers were 1.7 and 0.04%, respectively. Corresponding overall prevalences in steer samples were 27.2 and 0.6%. Campylobacter isolates were mostly C. jejuni and were tetracycline resistant. Eight Salmonella isolates were Salmonella Typhimurium that were either quad or penta resistant, most often to ampicillin, chloramphenicol, sulfamexathole, and tetracycline. Feedlot steers had greater odds of positive detection of Campylobacter (odds ratio, 8.5; confidence interval, 3.7, 19.5) than when grazing winter wheat. No breed effect was detected for fecal prevalence of these pathogens. DA - 2008/4// PY - 2008/4// DO - 10.4315/0362-028x-71.4.807 VL - 71 IS - 4 SP - 807-810 J2 - Journal of Food Protection LA - en OP - SN - 0362-028X UR - http://dx.doi.org/10.4315/0362-028x-71.4.807 DB - Crossref ER - TY - JOUR TI - Characterization of Salmonella enterica Serovar Agona Slaughter Isolates from the Animal Arm of the National Antimicrobial Resistance Monitoring System—Enteric Bacteria (NARMS): 1997 through 2003 AU - Douris, Aphrodite AU - Fedorka-Cray, Paula J. AU - Jackson, Charlene R. T2 - Microbial Drug Resistance AB - A total of 499 Salmonella enterica serovar Agona isolates from cattle, swine, chicken, and turkey samples were assayed for antimicrobial susceptibility and subtyped using pulsed-field gel electrophoresis (PFGE). Salmonella Agona isolates exhibited increased resistance to amoxicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, cephalothin, and chloramphenicol, and a single isolate was resistant to ceftriaxone. Multiple drug resistance (MDR; resistance ≥ 2 antimicrobials) was exhibited in 57% (n = 282/499) of the Salmonella Agona isolates and 22% (n = 111/499) of these Salmonella Agona isolates were resistant to five or more antimicrobials. PFGE patterns of 482 Salmonella Agona slaughter samples resulted in 165 unique patterns. Cluster analysis indicated that isolates indistinguishable by PFGE appeared to group according to antimicrobial resistance profiles. These data suggest that Salmonella Agona is increasing in prevalence in U.S. cattle presented for slaughter and should be further monitored. DA - 2008/3// PY - 2008/3// DO - 10.1089/mdr.2008.0787 VL - 14 IS - 1 SP - 55-63 J2 - Microbial Drug Resistance LA - en OP - SN - 1076-6294 1931-8448 UR - http://dx.doi.org/10.1089/mdr.2008.0787 DB - Crossref ER - TY - JOUR TI - Effect of Antimicrobial Dosage Regimen on Salmonella and Escherichia coli Isolates from Feeder Swine AU - Wagner, B. A. AU - Straw, B. E. AU - Fedorka-Cray, P. J. AU - Dargatz, D. A. T2 - Applied and Environmental Microbiology AB - A body of evidence exists that suggests that antimicrobial use in food animals leads to resistance in both pathogenic and commensal bacteria. This study focused on the impact of three different antimicrobial regimes (low-level continuous, pulse, and no antimicrobial) for two antimicrobials (chlortetracycline and tylosin) on the presence of Salmonella spp. and on the prevalence of antimicrobial resistance of both Salmonella spp. and nonspecific Escherichia coli in fecal samples from feeder swine. The prevalence of fecal samples positive for Salmonella spp. significantly decreased between the samples taken at feeder placement compared to samples taken when the animals were close to market weight. Differences in resistance of Salmonella spp. did not appear to be influenced by dosing treatment including the control. Analysis of antimicrobial resistance examining both susceptibility and resistance, as well as MIC outcomes, demonstrated that only resistance to cephalothin increased in E. coli under the pulse chlortetracycline treatment. These results suggest that the dosing regimes examined in this study did not lead to an increase in either the prevalence of Salmonella spp. or the prevalence of antimicrobial resistance in isolates of Salmonella spp. or E. coli. DA - 2008/1/25/ PY - 2008/1/25/ DO - 10.1128/aem.01132-07 VL - 74 IS - 6 SP - 1731-1739 J2 - Applied and Environmental Microbiology LA - en OP - SN - 0099-2240 UR - http://dx.doi.org/10.1128/aem.01132-07 DB - Crossref ER - TY - JOUR TI - First report of vatB and vgaB from Enterococcus gallinarum in the USA AU - Jackson, Charlene R. AU - Fedorka-Cray, Paula J. AU - Barrett, John B. AU - Hiott, Lari M. AU - Woodley, Tiffanie A. T2 - International Journal of Antimicrobial Agents AB - Cancer and hematological malignancies constitute major comorbidities in enterococcal infections, but little is known about the characteristics of enterococci affecting cancer patients. The aim of this study was to characterize 132 enterococcal clinical isolates obtained from cancer patients attending a Cancer Reference Center in Brazil between April 2013 and March 2014. Susceptibility to 17 antimicrobial agents was assessed by disk diffusion method. Resistance and virulence genes were investigated by PCR. Multilocus sequence typing (MLST) was performed for selected Enterococcus faecalis and Enterococcus faecium isolates. The predominant species was E. faecalis (108 isolates), followed by E. faecium (18), Enterococcus gallinarum (3), Enterococcus avium (2) and Enterococcus durans (1). Multidrug-resistant (MDR) isolates made up 44.7%, but all isolates were susceptible to fosfomycin, linezolid and glycopeptides. The most prevalent genes associated with erythromycin- and tetracycline-non susceptible isolates were erm(B) (47/71; 66.2%) and tet(M) (24/68; 35.3%), respectively. High-level resistance (HLR) to gentamicin was found in 22 (16.7%) isolates and 13 (59.1%) of them carried the aac(6′)-Ie-aph(2″)-Ia gene. HLR to streptomycin was detected in 34 (25.8%) isolates, of which 15 (44.1%) isolates had the ant(6′)-Ia gene. The most common virulence genes were gelE (48.9%), esp (30.5%) and asa1 (29.8%). MLST performed for 26 E. faecalis isolates revealed 18 different sequence-types (STs), with seven corresponding to novel STs (625, 626, 627, 628, 629, 630, and 635). On the other hand, nine of 10 E. faecium isolates analyzed by MLST belonged to a single clonal complex, comprised of mostly ST412, which emerged worldwide after mid-2000s, but also two novel STs (963 and 964). We detected major globally disseminated E. faecalis and E. faecium clonal complexes along with novel closely related STs, indicating the fitness and continuous evolution of these hospital-adapted lineages. DA - 2008/2// PY - 2008/2// DO - 10.1016/j.ijantimicag.2007.08.020 VL - 31 IS - 2 SP - 175-176 J2 - International Journal of Antimicrobial Agents LA - en OP - SN - 0924-8579 UR - http://dx.doi.org/10.1016/j.ijantimicag.2007.08.020 DB - Crossref ER - TY - JOUR TI - Urinary tract infections in female pigs. AU - Martineau, G AU - Almond, G T2 - CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources AB - Abstract This review is intended to provide an update on our current understanding of urinary tract infections (UTI) in female pigs. In general, there are two primary bacterial pathogens involved in these infections; however, several management and animal husbandry factors either contribute to or reduce the likelihood of infections. The precise molecular defence mechanisms in the porcine urinary bladder are poorly understood and extrapolation from other species may explain some of these mechanisms. Nevertheless, female pigs do possess inherent physiologic mechanisms for preventing the onset of UTI. To some extent, a 'country effect' appears to be involved with the significance of UTI in sows. Considerable efforts are made to diagnose and treat sows with UTI in France. In contrast, problems with UTI in sows tend to be overlooked in the USA. As shown in human medicine, perhaps the most significant challenge is the dilemma of asymptomatic bacteriuria. It is evident that considerable collaborative research is required to elucidate UTI in female pigs. DA - 2008/11/1/ PY - 2008/11/1/ DO - 10.1079/pavsnnr20083048 VL - 3 IS - 048 SP - 048, SN - 1749-8848 UR - http://dx.doi.org/10.1079/pavsnnr20083048 ER - TY - JOUR TI - Neuropathy Target Esterase Gene Mutations Cause Motor Neuron Disease AU - Rainier, Shirley AU - Bui, Melanie AU - Mark, Erin AU - Thomas, Donald AU - Tokarz, Debra AU - Ming, Lei AU - Delaney, Colin AU - Richardson, Rudy J. AU - Albers, James W. AU - Matsunami, Nori AU - Stevens, Jeff AU - Coon, Hilary AU - Leppert, Mark AU - Fink, John K. T2 - The American Journal of Human Genetics AB - The possibility that organophosphorus (OP) compounds contribute to motor neuron disease (MND) is supported by association of paraoxonase 1 polymorphisms with amyotrophic lateral sclerosis (ALS) and the occurrence of MND in OP compound-induced delayed neuropathy (OPIDN), in which neuropathy target esterase (NTE) is inhibited by organophosphorylation. We evaluated a consanguineous kindred and a genetically unrelated nonconsanguineous kindred in which affected subjects exhibited progressive spastic paraplegia and distal muscle wasting. Affected subjects resembled those with OPIDN and those with Troyer Syndrome due to SPG20/spartin gene mutation (excluded by genetic linkage and SPG20/spartin sequence analysis). Genome-wide analysis suggested linkage to a 22 cM homozygous locus (D19S565 to D19S884, maximum multipoint LOD score 3.28) on chromosome 19p13 to which NTE had been mapped (GenBank AJ004832 ). NTE was a candidate because of its role in OPIDN and the similarity of our patients to those with OPIDN. Affected subjects in the consanguineous kindred were homozygous for disease-specific NTE mutation c.3034A→G that disrupted an interspecies conserved residue (M1012V) in NTE's catalytic domain. Affected subjects in the nonconsanguineous family were compound heterozygotes: one allele had c.2669G→A mutation, which disrupts an interspecies conserved residue in NTE's catalytic domain (R890H), and the other allele had an insertion (c.2946_2947insCAGC) causing frameshift and protein truncation (p.S982fs1019). Disease-specific, nonconserved NTE mutations in unrelated MND patients indicates NTE's importance in maintaining axonal integrity, raises the possibility that NTE pathway disturbances contribute to other MNDs including ALS, and supports the role of NTE abnormalities in axonopathy produced by neuropathic OP compounds. The possibility that organophosphorus (OP) compounds contribute to motor neuron disease (MND) is supported by association of paraoxonase 1 polymorphisms with amyotrophic lateral sclerosis (ALS) and the occurrence of MND in OP compound-induced delayed neuropathy (OPIDN), in which neuropathy target esterase (NTE) is inhibited by organophosphorylation. We evaluated a consanguineous kindred and a genetically unrelated nonconsanguineous kindred in which affected subjects exhibited progressive spastic paraplegia and distal muscle wasting. Affected subjects resembled those with OPIDN and those with Troyer Syndrome due to SPG20/spartin gene mutation (excluded by genetic linkage and SPG20/spartin sequence analysis). Genome-wide analysis suggested linkage to a 22 cM homozygous locus (D19S565 to D19S884, maximum multipoint LOD score 3.28) on chromosome 19p13 to which NTE had been mapped (GenBank AJ004832 ). NTE was a candidate because of its role in OPIDN and the similarity of our patients to those with OPIDN. Affected subjects in the consanguineous kindred were homozygous for disease-specific NTE mutation c.3034A→G that disrupted an interspecies conserved residue (M1012V) in NTE's catalytic domain. Affected subjects in the nonconsanguineous family were compound heterozygotes: one allele had c.2669G→A mutation, which disrupts an interspecies conserved residue in NTE's catalytic domain (R890H), and the other allele had an insertion (c.2946_2947insCAGC) causing frameshift and protein truncation (p.S982fs1019). Disease-specific, nonconserved NTE mutations in unrelated MND patients indicates NTE's importance in maintaining axonal integrity, raises the possibility that NTE pathway disturbances contribute to other MNDs including ALS, and supports the role of NTE abnormalities in axonopathy produced by neuropathic OP compounds. Exposure to neurotoxic organophosphorous (OP) compounds of sufficient magnitude and duration causes neurodegeneration in humans, livestock, and laboratory animals.1Abou-Donia M.B. Organophosphorus ester-induced delayed neurotoxicity.Annu. Rev. Pharmacol. Toxicol. 1981; 21: 511-548Crossref PubMed Scopus (350) Google Scholar OP compound exposures occur in industrial or agricultural environments and as the consequences of terrorism, accidents, suicide attempts, or chemical warfare. Recent attention has focused on OP neurotoxicity after the use of Sarin in the 1995 Tokyo subway terrorist incident2Lotti M. Becker C.E. Aminoff M.J. Organophosphate polyneuropathy; pathogenesis and prevention.Neurology. 1984; 34: 658-662Crossref PubMed Google Scholar and because of concerns about persistent neurological effects after exposures to neurotoxic OP agents in the Gulf War. During the Prohibition era in the United States, consumption of Jamaica ginger extract (“Ginger Jake”) adulterated with triorthocresyl phosphate (TOCP) led to OP compound-induced delayed neuropathy (OPIDN) in an estimated 50,000 Americans.3Woolf A.D. Ginger Jake and the blues: A tragic song of poisoning.Vet. Hum. Toxicol. 1995; 37: 252-254PubMed Google Scholar, 4Morgan J.P. Penovich P. Jamaica ginger paralysis. Forty-seven-year follow-up.Arch. Neurol. 1978; 35: 530-532Crossref PubMed Scopus (97) Google Scholar, 5Morgan J.P. Tulloss T.C. The Jake Walk Blues. A toxicologic tragedy mirrored in American popular music.Ann. Intern. Med. 1976; 85: 804-808Crossref PubMed Scopus (31) Google Scholar Additional outbreaks of OPIDN causing paralysis of tens of thousands of individuals have occurred in Morocco, Fiji, and India due to consumption of cooking oil contaminated with lubricating oil (containing triorthocresyl phosphate).6Smith H.V. Spalding J.M. Outbreak of paralysis in Morocco due to ortho-cresyl phosphate poisoning.Lancet. 1959; 2: 1019-1021Abstract PubMed Scopus (106) Google Scholar, 7Taylor P. Anticholinesterase agents. Goodman and Gilman's Pharmacologic Basis of Therapeutics.Ninth Edition. McGraw Hill, New York1996Google Scholar, 8Sorokin M. Orthocresyl phosphate neuropathy: Report of an outbreak in Fiji.Med. J. Aust. 1969; 1: 506-508PubMed Google Scholar, 9Srivastava A.K. Das M. Khanna S.K. An outbreak of tricresyl phosphate poisoning in Calcutta, India.Food Chem. Toxicol. 1990; 28: 303-304Crossref PubMed Scopus (12) Google Scholar, 10Sarkar J.K. Outbreaks of paralystic disease in West Bengal due to tricresyl phosphate poisoning.J. Indian Med. Assoc. 1974; 63: 359-361PubMed Google Scholar, 11Mehta R.S. Dixit I.P. Khakharia S.J. Toxic neuropathy in Raipur due to triorthocresylphosphate (TOCP).J. Assoc. Physicians India. 1975; 23: 133-138PubMed Google Scholar Neurologic syndromes following OP toxicity are highly variable and depend on such factors as the specific OP compound, dose and duration of exposure, species, age, and various physiologic factors at the time of OP exposure. Whereas acute OP toxicity usually produces acute cholinergic crisis,7Taylor P. Anticholinesterase agents. Goodman and Gilman's Pharmacologic Basis of Therapeutics.Ninth Edition. McGraw Hill, New York1996Google Scholar acute cholinergic crises are often absent in OPIDN. In contrast, OPIDN often begins with sensory impairment, ataxia, weakness, muscle fasciculation, and hyporeflexia and may progress to complete flaccid paralysis followed by progressive spastic paraplegia.4Morgan J.P. Penovich P. Jamaica ginger paralysis. Forty-seven-year follow-up.Arch. Neurol. 1978; 35: 530-532Crossref PubMed Scopus (97) Google Scholar, 7Taylor P. Anticholinesterase agents. Goodman and Gilman's Pharmacologic Basis of Therapeutics.Ninth Edition. McGraw Hill, New York1996Google Scholar, 12Inoue N. Fujishiro K. Mori K. Matsuoka M. Triorthocresyl phosphate poisoning - A review of human cases.J. UOEH. 1988; 10: 433-442PubMed Google Scholar Neuropathologic analyses of OPIDN have shown distal degeneration of the longest central- and peripheral-nervous-system axons.1Abou-Donia M.B. Organophosphorus ester-induced delayed neurotoxicity.Annu. Rev. Pharmacol. Toxicol. 1981; 21: 511-548Crossref PubMed Scopus (350) Google Scholar, 12Inoue N. Fujishiro K. Mori K. Matsuoka M. Triorthocresyl phosphate poisoning - A review of human cases.J. UOEH. 1988; 10: 433-442PubMed Google Scholar Distal axon degeneration is also the primary neuropathologic feature of hereditary spastic paraplegia (HSP) (reviewed in13Fink J.K. Hereditary spastic paraplegia.in: Rimoin D. Connor J.M. Pyeritz R.E. Korf B.R. Emery and Rimoin's Principles and Practice of Medical Genetics. 5 ed. Churchill Livingstone Elsevier, Philadelphia2007: 2771-2801Google Scholar). Some complicated forms of HSP,13Fink J.K. Hereditary spastic paraplegia.in: Rimoin D. Connor J.M. Pyeritz R.E. Korf B.R. Emery and Rimoin's Principles and Practice of Medical Genetics. 5 ed. Churchill Livingstone Elsevier, Philadelphia2007: 2771-2801Google Scholar such as Troyer syndrome (MIM 275900), also exhibit lower-motor neuron involvement.14Patel H. Cross H. Proukakis C. Hershberger R. Bork P. Ciccarelli F.D. Patton M.A. McKusick V.A. Crosby A.H. SPG20 is mutated in Troyer syndrome, an hereditary spastic paraplegia.Nat. Genet. 2002; 31: 347-348Crossref PubMed Scopus (198) Google Scholar OPIDN pathogenesis involves neuropathy target esterase (NTE), a neuronal membrane protein, either through direct OP-induced inhibition of NTE or through generation of OP-NTE neurotoxic complexes (“aged NTE”). NTE's functions are incompletely understood. NTE is capable of hydrolyzing several intrinsic membrane lipids and thus may be a factor in determining the composition of neuronal membranes.15van Tienhoven M. Atkins J. Li Y. Glynn P. Human neuropathy target esterase catalyzes hydrolysis of membrane lipids.J. Biol. Chem. 2002; 277: 20942-20948Crossref PubMed Scopus (138) Google Scholar NTE's catalytic domain for esterase activity has been mapped to a 489 amino acid region between residues 727 and 1216.16Forshaw P.J. Atkins J. Ray D.E. Glynn P. The catalytic domain of human neuropathy target esterase mediates an organophosphate-sensitive ionic conductance across liposome membranes.J. Neurochem. 2001; 79: 400-406Crossref PubMed Scopus (12) Google Scholar Participation of NTE in a cell-signaling pathway controlling interactions between neurons and accessory glial cells in the developing nervous system has also been proposed.17Glynn P. Neuropathy target esterase.Biochem. J. 1999; 344: 625-631Crossref PubMed Scopus (107) Google Scholar, 18Glynn P. Neural development and neurodegeneration: Two faces of neuropathy target esterase.Prog. Neurobiol. 2000; 61: 61-74Crossref PubMed Scopus (107) Google Scholar We studied a consanguineous family of Ashkenazi Jewish ancestry (Figure 1A) and a nonconsanguineous family of northern European ancestry (Figure 1B) in which affected subjects developed childhood onset of insidiously progressive lower-extremity spastic weakness and progressive wasting of distal upper- and lower-extremity muscles. Electrophysiologic studies were consistent with a motor axonopathy affecting upper and lower extremities. Magnetic resonance imaging demonstrated spinal cord atrophy, particularly in the thoracic region. The affected phenotype in each family conformed both to OPIDN and to “Troyer syndrome,” an autosomal-recessive form of HSP associated with distal muscle wasting.14Patel H. Cross H. Proukakis C. Hershberger R. Bork P. Ciccarelli F.D. Patton M.A. McKusick V.A. Crosby A.H. SPG20 is mutated in Troyer syndrome, an hereditary spastic paraplegia.Nat. Genet. 2002; 31: 347-348Crossref PubMed Scopus (198) Google Scholar, 19Cross H.E. McKusick V.A. The Troyer syndrome. A recessive form of spastic paraplegia with distal muscle wasting.Arch. Neurol. 1967; 16: 473-485Crossref PubMed Scopus (93) Google Scholar Troyer syndrome patients variably exhibit additional neurologic and systemic abnormalities (delayed milestone acquisition, skeletal abnormalities, and cerebellar, extrapyramidal, and cognitive impairment). These features were not observed in our patients. The University of Michigan Institutional Review Board approved this study. We analyzed the SPG20/spartin gene (MIM 607111) coding sequence, mutations in which cause Troyer syndrome (SPG20 HSP),14Patel H. Cross H. Proukakis C. Hershberger R. Bork P. Ciccarelli F.D. Patton M.A. McKusick V.A. Crosby A.H. SPG20 is mutated in Troyer syndrome, an hereditary spastic paraplegia.Nat. Genet. 2002; 31: 347-348Crossref PubMed Scopus (198) Google Scholar, 19Cross H.E. McKusick V.A. The Troyer syndrome. A recessive form of spastic paraplegia with distal muscle wasting.Arch. Neurol. 1967; 16: 473-485Crossref PubMed Scopus (93) Google Scholar and found no mutations in the index (consanguineous) family (data not shown), and we excluded the SPG2014Patel H. Cross H. Proukakis C. Hershberger R. Bork P. Ciccarelli F.D. Patton M.A. McKusick V.A. Crosby A.H. SPG20 is mutated in Troyer syndrome, an hereditary spastic paraplegia.Nat. Genet. 2002; 31: 347-348Crossref PubMed Scopus (198) Google Scholar locus in the consanguineous family by genetic-linkage analysis (Table 1). Two-point linkage analyses for exclusion mapping were performed with the MLINK subroutine of the LINKAGE program20Lathrop G.M. Lalouel J. Julient C. Ott J. Multipoint linkage analysis in humans: Detection of linkage and estimation of recombination.Am. J. Hum. Genet. 1985; 37: 482-498PubMed Google Scholar with an autosomal-recessive model of disease inheritance and a disease allele frequency of 0.001. We assigned a genetic penetrance of 0.90 for LOD-score calculations.Table 1Genetic-Linkage Analysis Excludes Chromosome 13q12.3MarkerLOD θ = 00.010.020.030.040.05D13S1841−3.4−1.23−0.94−0.77−0.66−0.57D13S1842−2.19−1.19−0.92−0.76−0.64−0.56D13S1851−3.4−1.23−0.94−0.77−0.66−0.57D13S1843−3.4−1.23−0.94−0.77−0.66−0.57D13S2190.080.070.070.060.060.06D13S1844−2.19−1.19−0.92−0.76−0.64−0.56Genetic-linkage analysis excludes SPG20 “Troyer syndrome” locus on chromosome 13q12.3. Open table in a new tab Genetic-linkage analysis excludes SPG20 “Troyer syndrome” locus on chromosome 13q12.3. We then initiated genome-wide linkage analysis (Figure 2) in the larger, consanguineous family with ten subjects (three living affected subjects, seven living unaffected subjects, no spouses of descendants) and in the second smaller, nonconsanguineous family. Marrying-in spouses were asymptomatic and had no evidence of similar neurologic disorders in their families. Maximum-likelihood estimates of marker allele frequencies were estimated from all 15 subjects in both families. The average number of alleles was 4 (range = 3–7), and the average observed heterozygosity of these markers in the families was 0.64 (range = 0.32–0.84). We analyzed 400 polymorphic microsatellite markers spaced ∼10 cM apart (ABI MD-10 Linkage Mapping Set), of which 164 markers heterozygous for at least one affected individual excluded 41% of the genome. Six of these markers were homozygous for all affected individuals in the consanguineous family. We analyzed adjacent markers for each homozygous marker. Only markers adjacent to D19S209 yielded an extended linked haplotype that spanned 22 cM between D19S565 and D19S884 on chromosome 19p13 (Figure 1A). In the smaller, nonconsanguineous family, these same markers also resulted in haplotype sharing in affected subjects consistent with genetic linkage of this region (Figure 1B). The positive chromosome 19 region was analyzed with multipoint methods to incorporate information from all 27 informative markers. The GeneHunter program21Kruglyak L. Daly M.J. Reeve-Daly M.P. Lander E.S. Parametric and nonparametric linkage analysis: A unified multipoint approach.Am. J. Hum. Genet. 1996; 58: 1347-1363PubMed Google Scholar was used to obtain exact multipoint LOD scores and multipoint nonparametric linkage (NPL) scores. The NPL score is based on allele sharing identical by descent (IBD) and does not assume a transmission model. The NPL analysis allows an assessment of the robustness of the result to model misspecification. Because of the size and complexity of the largest pedigree, it had to be trimmed for analysis in GeneHunter. Therefore, analyses were also performed with SimWalk2.22Sobel E. Lange K. Descent graphs in pedigree analysis: Applications to haplotyping, location scores, and marker sharing statistics.Am. J. Hum. Genet. 1996; 58: 1323-1337PubMed Google Scholar, 23Sobel E. Sengul H. Weeks D.E. Multipoint estimation of identity-by-descent probablilities at arbitrary positions among marker loci on general pedigrees.Hum. Hered. 2001; 52: 121-131Crossref PubMed Scopus (145) Google Scholar, 24Sobel E. Papp J.C. Jange K. Detection and integration of genotyping errors in statistical genetics.Am. J. Hum. Genet. 2002; 70: 496-508Abstract Full Text Full Text PDF PubMed Scopus (282) Google Scholar SimWalk2 uses Markov chain Monte Carlo (MCMC) and simulated annealing methods to perform multipoint analyses, allowing computation on large, complex pedigrees. Although the method is not exact, all pedigree members could be included in the analysis. We performed parametric LOD score and NPL analyses in SimWalk2. For each analysis, we set the “parallel runs” flag to generate two simulated annealing runs of the data. The runs produced a maximum score at the same location with the same scores within rounding error. p values given by SimWalk2 were calculated with 10,000 simulations. For nonparametric analyses, p values were used to generate the reported scores (score = −log10 [p value]). Multipoint LOD score and nonparametric analyses of the two families clearly mirrored the shared haplotype region (Figure 2). With all methods (model-based parametric analysis or nonparametric analysis with either GeneHunter or SimWalk2) used, the maximum LOD score occurred near marker D19S869. Maximum multipoint parametric LOD scores were 2.58 with GeneHunter and 3.82 with SimWalk2. The maximum NPL scores were 3.30 with GeneHunter and 3.36 with SimWalk2. The p value associated with the GeneHunter statistics is 0.002; the simulation-based p value for the SimWalk2 statistics is 0.0004. The index family contributed most to these scores (3.28 and 3.09 for the parametric and NPL SimWalk2 analyses, respectively, and 2.07 and 3.07 for the GeneHunter parametric and NPL analyses, respectively). It is important to note that the second small, nonconsanguineous family supported this peak, with scores close to the maximum expected for an affected-sibling-pair family. To further characterize the region on chromosome 19 and to fully characterize all shared chromosomal regions, we carried out high-density SNP typing with the Affymetrix 250K NspI chips on all family members for whom DNA was available: 12 subjects in the consanguineous family (including all three affected individuals and two additional unaffected subjects ascertained after the initial microsatellite linkage analysis had been performed) and five subjects in the nonconsanguineous family. We analyzed the SNP data by looking for regions of autozygosity in the entire genome with the Find Autozygous Regions program in the GeneSpring GT analysis package (Agilent). The results of this analysis are shown in Figure 3; they clearly show a single major autozygosity region on chromosome 19 in the three affected individuals. This 4.7 megabase region is defined by the map location from 2784431 to 7541689 bp (NCBI build 35). This region corresponds to the same region identified by the microsatellite markers. The region includes the interesting candidate gene neuropathy target esterase (NTE) (GenBank AJ004832 ), whose location on chromosome 19 is from 7505075—7532647 bp (UCSC Genome Browser on NCBI build 35). The NTE gene was an obvious candidate because of its role in OPIDN and the similarity of symptoms seen in our patients to those reported for OPIDN. Analysis of NTE's coding sequence in the index family showed that each affected subject was homozygous for (and each obligate carrier heterozygous for) substitution of guanine for adenine at NTE cDNA 3034. This mutation was absent in 105 control subjects (data not shown) and caused substitution of valine for methionine at amino acid position 1012 (M1012V). This mutation disrupts an interspecies conserved residue within NTE's catalytic domain (Figure 4C).25Lush M.J. Li Y. Read D.J. Willis A.C. Glynn P. Neuropathy target esterase and a homologous Drosophila neurodegeneration-associated mutant protein contain a novel domain conserved from bacteria to man.Biochem. J. 1998; 332: 1-4Crossref PubMed Scopus (122) Google Scholar Mouse, Drosophila, and C. elegans species all contain M at residue 1012. Polyphen analysis of M1012V mutation gave a PSIC profile-difference score of 2.590 predicting that the mutation would be damaging. On the other hand, SIFT analysis predicted that M1012V substitution would be tolerated. We then analyzed the NTE coding sequence in the second, unrelated, nonconsanguineous family (Figure 1B). Analysis of NTE's coding sequence in affected subjects in this second family showed that they were compound heterozygotes for two NTE mutations. Like the mutation in the index family, these mutations also occurred within NTE's catalytic domain. One allele had a 2669G→A mutation corresponding to an R890H substitution in NTE's catalytic domain. Human and mouse contain R at residue 890, whereas Drosophila and C.elegans contain K at residue 890. Polyphen analysis (PSIC profile-difference score = 0.184) and SIFT analysis predicted that R890H would be a tolerated substitution. The other allele had a four base pair insertion (NTE mRNA position 2946) that caused a frameshift and protein truncation after residue 1019. The truncated protein is predicted to be missing the last 235 residues of NTE's catalytic domain (which extends from amino acid position 727 to position 1216). These mutations were present separately in each carrier parent and absent in 105 control subjects and the unaffected sibling. In summary, we identified homozygous and compound heterozygous NTE mutations in subjects from two unrelated families with autosomal-recessive progressive spastic paraplegia associated with distal upper- and lower-extremity wasting. Because of its clinical similarity with SPG20 HSP (Troyer syndrome), we reserved for this disease the next-available HSP locus designation (SPG39) and refer to this disorder as NTE-related Motor Neuron Disorder (NTE-MND). These NTE mutations are disease-specific and considered pathogenic for several reasons. First, they were present in affected subjects in two unrelated kindreds and absent in control subjects. Second, they affect amino acid residues within NTE's catalytic domain. Third, NTE plays a central role in OPIDN, an upper- and lower-motor neuron disorder whose symptoms bear a striking resemblance to those exhibited by our NTE-MND patients. Two mechanisms for NTE involvement in OPIDN have been proposed. The first mechanism proposes that neurotoxicity is the consequence of OP-induced inhibition of NTE. The second mechanism proposes that OP-NTE complexes are toxic. These proposed mechanisms are not mutually exclusive because OP interaction with NTE could both be toxic as well as interfere with NTE activity. The fact that identified NTE mutations in NTE-MND subjects disturb NTE's catalytic domain suggests that they could result in altered NTE activity in vivo. This suggests that modified NTE activity alone could be sufficient to cause corticospinal tract and peripheral motor axonopathy. Nonetheless, it is also possible that the NTE mutations identified result in neurotoxic or “aged” NTE. Observations that NTE mutations are associated with progressive upper- and lower-motor neuron disease indicate the importance of NTE in maintaining the integrity of corticospinal tract and peripheral motor axons. The finding that NTE mutations underlie corticospinal tract and peripheral motor axon degeneration raises the possibility that other NTE polymorphisms (or genetic variation in factors that regulate or interact with NTE) could contribute to other motor neuron disorders including amyotrophic lateral sclerosis (ALS) and primary lateral sclerosis (PLS). We have shown that two NTE mutations are the likely cause of autosomal-recessive motor neuron disease that closely resembles OPIDN. These NTE mutations were sufficient to cause the disorder even in the absence of apparent exposure to neurotoxic OP compounds. It will be important to determine whether other polymorphisms in NTE and/or proteins with which it interacts influence the susceptibility to OP-induced neurologic disease. Our findings, together with the recently identified association of PON1 polymorphisms with ALS,26Slowik A. Tomik B. Wolkow P.P. Partyka D. Turaj W. Malecki M.T. Pera J. Dziedzic T. Szczudlik A. Figlewicz D.A. Paraoxonase gene polymorphisms and sporadic ALS.Neurology. 2006; 67: 766-770Crossref PubMed Scopus (72) Google Scholar further support the possibility that neurotoxic OP compounds contribute to motor neuron disease. This research is supported by grants from the University of Michigan Institute of Gerontology (to S.R.); and the Veterans Affairs Merit Review, the National Institutes of Health (National Institute of Neurological Disorders and Stroke [NINDS] R01-NS053917 and R01-NS045163), the Spastic Paraplegia Foundation, and the National Organization for Rare Disorders (to J.K.F.). Those funding this study did not play any role in design and conduct of the study, in the collection, analysis, and interpretation of the data, or in the preparation, review, or approval of the manuscript. We are grateful for the Elderly Subjects Program of the University of Michigan Institute of Gerontology through which control subjects were ascertained, the technical assistance of Shalini Guduri, the expert secretarial assistance of Lynette Girbach, and the participation of research subjects and their families without whom this investigation would not be possible. Download .xls (.1 MB) Help with xls files Document S1. Table of Genescan Data The URL for online data listed herein is as follows:Online Mendelian Inheritance in Man, http://www.ncbi.nlm.nih.gov/Omim/. DA - 2008/3// PY - 2008/3// DO - 10.1016/j.ajhg.2007.12.018 VL - 82 IS - 3 SP - 780-785 J2 - The American Journal of Human Genetics LA - en OP - SN - 0002-9297 UR - http://dx.doi.org/10.1016/j.ajhg.2007.12.018 DB - Crossref ER - TY - CHAP TI - Mechanisms of Cell Death AU - Law, Mac AU - Elmore, Susan T2 - Molecular and Biochemical Toxicology AB - This chapter contains sections titled: Introduction How Cells/Tissues React to “Stress” Cell Injury and Cell Death Morphology of Cell Injury and Cell Death Apoptosis Suggested Reading PY - 2008/7/11/ DO - 10.1002/9780470285251.ch16 SP - 287-318 OP - PB - John Wiley & Sons, Inc. SN - 9780470285251 9780470102114 UR - http://dx.doi.org/10.1002/9780470285251.ch16 DB - Crossref ER - TY - JOUR TI - Antimicrobial Susceptibility of Foodborne Pathogens in Organic or Natural Production Systems: An Overview AU - Jacob, Megan E. AU - Fox, James Trent AU - Reinstein, Shelby L. AU - Nagaraja, T.G. T2 - Foodborne Pathogens and Disease AB - Organic and natural food production systems are increasing in popularity, at least partially because consumers perceive that these niche markets provide healthier and safer food products. One major difference between these niche markets and conventional production systems is the use of antimicrobials. Because antimicrobial agents exert selective pressures for antimicrobial resistance, relating antimicrobial susceptibility of foodborne bacteria to niche market production systems is of interest. Other differences between production systems might also influence the susceptibility of foodborne pathogens. The objective of this review is to compare the impact of food animal production systems on the antimicrobial susceptibility of common foodborne bacterial pathogens. Studies comparing the susceptibility of such pathogens were diverse in terms of geographic location, procedures, species of bacteria, and antimicrobials evaluated; thus, it was difficult to draw conclusions. The literature is highly variable in terms of production type and practices and susceptibility associations, although few studies have compared truly organic and conventional practices. When statistical associations were found between production type and minimum inhibitory concentrations or percentage of isolates resistant for a particular pathogen, the isolates from conventionally reared animals/products were more commonly resistant than the comparison group (organic, antibiotic free, etc.). Therefore, further studies are needed to better assess public health consequences of antimicrobial resistance and food animal production systems, specifically organic or natural versus conventional. DA - 2008/12// PY - 2008/12// DO - 10.1089/fpd.2008.0095 VL - 5 IS - 6 SP - 721-730 J2 - Foodborne Pathogens and Disease LA - en OP - SN - 1535-3141 1556-7125 UR - http://dx.doi.org/10.1089/fpd.2008.0095 DB - Crossref ER - TY - JOUR TI - Niche Marketing Production Practices for Beef Cattle in the United States and Prevalence of Foodborne Pathogens AU - Fox, J. Trent AU - Reinstein, Shelby AU - Jacob, Megan E. AU - Nagaraja, T.G. T2 - Foodborne Pathogens and Disease AB - Niche-marketed food products are rapidly gaining market share in today's society. Consumers are willing to pay premium prices for food perceived to be safer, healthier, more nutritious, and better tasting than conventional food. This review outlines typical production practices for niche-market beef production systems in the United States and compares prevalence estimates of foodborne pathogens in animals and produce from conventional and niche-market production systems. The two main niches for food animal production are organic and natural productions. Organic and natural beef productions are becoming increasingly popular and there is high consumer demand. Two major differences between conventional beef production systems and niche-market production systems (natural and organic) are in the use of antimicrobials and growth-promoting hormones. The impacts of these production systems on foodborne pathogens in beef cattle are variable and often data are nonexistent. Studies directly comparing conventional and niche-market production systems for dairy, swine, poultry, and produce have observed that the prevalence of foodborne pathogens was seldom statistically different between production systems, but when differences were observed, prevalence was typically greater for the niche-market production systems than the conventional production system. The published literature suggests that the perception of niche-marketed food products being safer and healthier for consumers with regard to foodborne pathogens may not be justified. DA - 2008/10// PY - 2008/10// DO - 10.1089/fpd.2008.0094 VL - 5 IS - 5 SP - 559-569 J2 - Foodborne Pathogens and Disease LA - en OP - SN - 1535-3141 1556-7125 UR - http://dx.doi.org/10.1089/fpd.2008.0094 DB - Crossref ER - TY - JOUR TI - Effects of feeding wet corn distillers grains with solubles with or without monensin and tylosin on the prevalence and antimicrobial susceptibilities of fecal foodborne pathogenic and commensal bacteria in feedlot cattle1 AU - Jacob, M. E. AU - Fox, J. T. AU - Narayanan, S. K. AU - Drouillard, J. S. AU - Renter, D. G. AU - Nagaraja, T. G. T2 - Journal of Animal Science AB - Distillers grains, a coproduct of ethanol production from cereal grains, are composed principally of the bran, protein, and germ fractions and are commonly supplemented in ruminant diets. The objective of this study was to assess the effect of feeding wet distillers grains with solubles (WDGS) and monensin and tylosin on the prevalence and antimicrobial susceptibilities of fecal foodborne and commensal bacteria in feedlot cattle. Cattle were fed 0 or 25% WDGS in steam-flaked corn-based diets with the addition of no antimicrobials, monensin, or monensin and tylosin. Fecal samples were collected from each animal (n = 370) on d 122 and 136 of the 150-d finishing period and cultured for Escherichia coli O157. Fecal samples were also pooled by pen (n = 54) and cultured for E. coli O157, Salmonella, commensal E. coli, and Enterococcus species. Antimicrobial resistance was assessed by determining antimicrobial susceptibilities of pen bacterial isolates and quantifying antimicrobial resistance genes in fecal samples by real-time PCR. Individual animal prevalence of E. coli O157 in feces collected from cattle fed WDGS was greater (P < 0.001) compared with cattle not fed WDGS on d 122 but not on d 136. There were no treatment effects on the prevalence of E. coli O157 or Salmonella spp. in pooled fecal samples. Antimicrobial susceptibility results showed Enterococcus isolates from cattle fed monensin or monensin and tylosin had greater levels of resistance toward macrolides (P = 0.01). There was no effect of diet or antimicrobials on concentrations of 2 antimicrobial resistance genes, ermB or tetM, in fecal samples. Results from this study indicate that WDGS may have an effect on the prevalence of E. coli O157 and the concentration of selected antimicrobial resistance genes, but does not appear to affect antimicrobial susceptibility patterns in Enterococcus and generic E. coli isolates. DA - 2008/5/1/ PY - 2008/5/1/ DO - 10.2527/jas.2007-0091 VL - 86 IS - 5 SP - 1182-1190 LA - en OP - SN - 0021-8812 1525-3163 UR - http://dx.doi.org/10.2527/jas.2007-0091 DB - Crossref KW - antimicrobial feed additive KW - antimicrobial susceptibility KW - cattle KW - distillers grain KW - Escherichia coli O157 ER - TY - JOUR TI - Feeding Supplemental Dried Distiller’s Grains Increases Faecal Shedding of Escherichia coli O157 in Experimentally Inoculated Calves AU - Jacob, M. E. AU - Parsons, G. L. AU - Shelor, M. K. AU - Fox, J. T. AU - Drouillard, J. S. AU - Thomson, D. U. AU - Renter, D. G. AU - Nagaraja, T. G. T2 - Zoonoses and Public Health AB - Summary Escherichia coli O157 is an important foodborne pathogen and asymptomatic cattle serve as major reservoirs for human infection. We have shown a positive association between feeding distiller’s grains and E. coli O157 prevalence in feedlot cattle. The objective of this study was to determine the effect of feeding dried distiller’s grain (DDG) on faecal shedding of E. coli O157 in calves experimentally inoculated with E. coli O157. Holstein calves (five per treatment group), fed steam‐flaked corn‐based high‐grain diets supplemented with 0% (control) or 25% DDG, were orally inoculated with a five‐strain mixture (6 × 10 9 CFU/calf) of nalidixic acid‐resistant ( Nal R ) E. coli O157. Faecal samples were taken three times per week for 6 weeks to determine the prevalence and concentration of Nal R E. coli O157. At the end of the study (day 43), calves were euthanized and necropsied. Ruminal, caecum, colon, and rectal contents, and rectoanal mucosal swab (RAMS) samples were collected at necropsy to determine Nal R E. coli O157 concentration. There was a trend for an interaction between treatment and faecal sampling day. The concentration of Nal R E. coli O157 in the faeces was significantly higher in faecal samples from calves fed DDG compared with control calves on days 35, 37, 39 and 42. At necropsy, the concentration of Nal R E. coli O157 was higher in the caecum ( P = 0.01), colon ( P = 0.03) and rectum ( P = 0.01) from calves fed DDG compared with control animals. The number of sites at necropsy positive for Nal R E. coli O157 was higher in calves fed DDG compared with calves in the control treatment ( P < 0.001). Our results indicate that E. coli O157 gut persistence and faecal prevalence increased in calves fed DDG, which potentially have important implications for food safety. DA - 2008/4// PY - 2008/4// DO - 10.1111/j.1863-2378.2008.01115.x VL - 55 IS - 3 SP - 125-132 J2 - Zoonoses Public Health LA - en OP - SN - 1863-1959 1863-2378 UR - http://dx.doi.org/10.1111/j.1863-2378.2008.01115.x DB - Crossref ER - TY - JOUR TI - Deletion of the Marek’s disease virus UL41 gene (vhs) has no measurable effect on latency or pathogenesis AU - Gimeno, Isabel AU - Silva, Robert F. T2 - Virus Genes DA - 2008/6// PY - 2008/6// DO - 10.1007/s11262-008-0215-3 VL - 36 IS - 3 SP - 499-507 J2 - Virus Genes LA - en OP - SN - 0920-8569 1572-994X UR - http://dx.doi.org/10.1007/s11262-008-0215-3 DB - Crossref KW - herpesvirus KW - poultry KW - avian KW - mutant KW - pathogenesis KW - MDV ER - TY - CHAP TI - CD4+CD25+ Regulatory T Cells in Viral Infections AU - Tompkins, Wayne A. AU - Tompkins, Mary B. AU - Mexas, Angela M. AU - Fogle, Jonathan E. T2 - Regulatory T Cells and Clinical Application PY - 2008/// DO - 10.1007/978-0-387-77909-6_22 SP - 407-422 OP - PB - Springer US SN - 9780387779089 9780387779096 UR - http://dx.doi.org/10.1007/978-0-387-77909-6_22 DB - Crossref ER - TY - JOUR TI - Detection of Mycoplasma pulmonis in Laboratory Rats and Technicians AU - Ferreira, J. B. AU - Yamaguti, M. AU - Marques, L. M. AU - Oliveira, R. C. AU - Neto, R. L. AU - Buzinhani, M. AU - Timenetsky, J. T2 - Zoonoses and Public Health AB - Summary Five species of mycoplasma are associated with several rat diseases. Mycoplasma pulmonis is the most important and most studied, possibly causing disease in rats and undermining the validity of laboratory experiments. M. pulmonis was isolated in 144/240 laboratory rats and identified by PCR in 155/240. This species was also detected in 12 human individuals (technicians of a laboratory animal house hold) in contact with these rats. The results were confirmed by sequencing of DNA products. Mycoplasma species are host specific; however, M. pulmonis was identified in humans, suggesting a case of unspecific colonization. Statistical analysis shows a greater risk for M. pulmonis colonizing individuals who are exposed to infected rats in animal facilities than individuals who do not. The detection of M. pulmonis in humans indicates a new status for this mollicute mycoplasmas in animal‐holding facilities. DA - 2008/6// PY - 2008/6// DO - 10.1111/j.1863-2378.2008.01122.x VL - 55 IS - 5 SP - 229-234 J2 - Zoonoses Public Health LA - en OP - SN - 1863-1959 1863-2378 UR - http://dx.doi.org/10.1111/j.1863-2378.2008.01122.x DB - Crossref ER - TY - CONF TI - Suvaxyn® Circo ameliorates PCV2-associated lymphoid lesions and PMWS-associated mortality AU - Segalés, J AU - Urniza, A AU - Alegre, A AU - Crisci, E AU - Pérez, M AU - Nofrarı́as, M AU - Balasch, M AU - Xu, Z AU - Chu, HJ AU - Plana-Duran, J C2 - 2008/// C3 - 20th INTERNATIONAL PIG VETERINARY SOCIETY CONGRESS DA - 2008/// SP - 18 ER - TY - CONF TI - IMMUNOMODULATION IN PRRS MLV VACCINATION AU - Fraile, L AU - Crisci, E AU - Gimeno, M AU - Diaz, I AU - Mateu, E AU - Montoya, M C2 - 2008/// C3 - 20th INTERNATIONAL PIG VETERINARY SOCIETY CONGRESS DA - 2008/// SP - 21 ER - TY - JOUR TI - A proposal on porcine circovirus type 2 (PCV2) genotype definition and their relation with postweaning multisystemic wasting syndrome (PMWS) occurrence. AU - Grau-Roma, L AU - Crisci, E AU - Sibila, M AU - López-Soria, S AU - Nofrarias, M AU - Cortey, M AU - Fraile, L AU - Olvera, A AU - Segalés, J T2 - Veterinary microbiology AB - Porcine circovirus type 2 (PCV2) is the essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). Despite first sequencing studies did not find any association between PCV2 sequences and PMWS occurrence, recent works have suggested the opposite. In the present study, 87 open reading frame 2 (ORF2) sequences obtained from pigs with different clinical conditions and coming from farms with different PMWS status were analyzed. Results further confirmed the existence of two genogroups and the definition of two PCV2 genotypes (1 and 2) is proposed. All sequences included in genotype 1 came from pigs from PMWS affected farms, while all sequences obtained from non-PMWS affected farms corresponded to genotype 2. Moreover, infection of single pigs from PMWS affected farms harbouring both genotypes is described. Present results suggest that PCV2 genotype 1 may potentially be more pathogenic than PCV2 genotype 2. DA - 2008/4// PY - 2008/4// DO - 10.1016/j.vetmic.2007.09.007 VL - 128 IS - 1-2 SP - 23–35 UR - http://europepmc.org/abstract/med/17976930 KW - porcine circovirus type 2 (PCV2) KW - postweaning multisystemic wasting syndrome (PMWS) KW - genotype KW - phylogeny KW - epidemiology KW - pathogenicity ER - TY - JOUR TI - Phenotypic and functional characterisation of porcine CD4+CD25high regulatory T cells AU - Käser, T. AU - Gerner, W. AU - Hammer, S.E. AU - Patzl, M. AU - Saalmüller, A. T2 - Veterinary Immunology and Immunopathology AB - Over the last years regulatory T cells (Tregs) were defined as CD4(+)CD25(+) T lymphocytes expressing the transcription factor Foxp3 (Forkhead Box P3) with the ability to downregulate various immune responses. In swine the existence of CD4(+)CD25(+) T lymphocytes was described before but nothing is known about the function of this minor cell population to date. Therefore, we studied porcine CD4(+)CD25(+) T cells with regard to major attributes of murine and human Tregs: their phenotype concerning the expression of several T-cell specific antigens, including Foxp3, their IL-10 production and their suppressive capacity. Our results revealed that porcine CD4(+)CD25(+) T cells with high CD25 expression count for about 2-9% of the CD4(+) T-cell subset. They demonstrate a strong Foxp3 expression, a heterogeneous CD45RC-, CD8alpha- and MHC-II-defined phenotype and a moderate IL-10 production. Co-cultivation of increasing numbers of CD4(+)CD25(high) T cells with a constant number of CD4(+)CD25(-) responder-T cells caused a decrease in proliferation of the entire culture. This demonstrates the suppressive capacity of the CD4(+)CD25(high) T-cell subset and - together with their Foxp3 expression - the existence of porcine Tregs. DA - 2008/// PY - 2008/// DO - 10.1016/j.vetimm.2007.08.002 VL - 122 IS - 1-2 SP - 153-158 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-39449127746&partnerID=MN8TOARS KW - regulatory T cells KW - swine KW - foxp3 KW - T lymphocytes ER - TY - JOUR TI - Detection of intracellular antigens in porcine PBMC by flow cytometry: A comparison of fixation and permeabilisation reagents T2 - Veterinary Immunology and Immunopathology AB - The staining of intracellular antigens is a standard method in flow cytometry but still only occasionally used for immunologic studies in veterinary species. In order to improve information about suitable fixation/permeabilisation protocols in porcine lymphocytes we tested six fixation/permeabilisation variants for the staining of different intracellular antigens: CD79α, perforin, interferon-γ and the cell cycle associated protein Ki-67. For the fixation/permeabilisation procedure, four commercial kits from BD Biosciences, ADG Bio Research, Dako and AbD Serotec were tested in comparison to non-commercial fixation solutions of formaldehyde together with either saponin or Tween-20 for permeabilisation. Perforin and Ki-67 expressing cells were optimally stained only with permeabilisation reagents containing saponin, which includes the BD kit. In contrast, labelling of CD79α and IFN-γ was possible with all investigated fixation/permeabilisation variants; however, here saponin and Tween protocols induced a higher degree of unspecific binding. Also, scatter properties of cells treated with saponin were worse compared to samples prepared with the kits from ADG, Dako and Serotec. Simultaneous staining of cell surface antigens was not negatively affected by any of the fixation/permeabilisation variants. A universal recommendation for a single fixation/permeabilisation strategy could not be deduced from our data but our work provides a useful guideline for optimal staining of common intracellular antigens analyzed in swine lymphocytes. DA - 2008/// PY - 2008/// DO - 10.1016/j.vetimm.2007.09.019 VL - 121 IS - 3-4 SP - 251-259 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-38549177571&partnerID=MN8TOARS KW - intracellular antigen KW - swine KW - fixation KW - permeabilisation KW - flow cytometry ER - TY - JOUR TI - Detection of Foxp3 protein expression in porcine T lymphocytes AU - Käser, T. AU - Gerner, W. AU - Hammer, S.E. AU - Patzl, M. AU - Saalmüller, A. T2 - Veterinary Immunology and Immunopathology AB - Regulatory T cells (Tregs) are potent regulators of various immune reactions. Due to the lack of Treg-specific markers their analysis had often been challenging until the discovery of the transcription factor Forkhead-box p3 (Foxp3) which serves as this highly demanded marker. So far, antibodies staining human and murine Foxp3 have been developed. This study describes the analysis of four commercially available anti-Foxp3 antibodies for reactivity with their specific antigen in cells derived from porcine lymphoid tissues. One out of the four antibodies showed selective reactivity with porcine CD25(+) T lymphocytes. The intracellular antigen was expressed on a small subset of CD25(dim) cells and the majority of the CD25(high) positive T-cell subpopulation. Intracellular antigen positive cells showed a heterogeneous expression of other leukocyte differentiation antigens. The majority belonged to the CD4(+)CD8(+) T-lymphocyte subpopulation, but were also found in the CD4(+)CD8(-) subpopulation. Another small minority was included in the CD4(-)CD8(+) T-lymphocyte subpopulation. Additionally, a small fraction of the putative Foxp3(+) cells showed an expression of MHC-II molecules. These staining patterns in three and four colour flow cytometry analyses indicated that the cells detected by a rat anti-mouse/rat-Foxp3 antibody expressed the porcine Foxp3. The expression of the putative Foxp3 protein in distinct leukocyte subsets was confirmed by molecular analysis of Foxp3 mRNA expression. Using Western blot analysis specific protein bands could only be detected in fractions that also exhibited the corresponding Foxp3 mRNA expression. These experiments also revealed that the antibody recognized a single chain protein with a molecular mass of about 45kDA similar to Foxp3 described for other species. In summary, these data strongly indicate the reactivity of this antibody with porcine Foxp3. Thereby, this rat anti-mouse/rat Foxp3 antibody presents a powerful tool for the identification of porcine regulatory T cells. DA - 2008/// PY - 2008/// DO - 10.1016/j.vetimm.2008.05.007 VL - 125 IS - 1-2 SP - 92-101 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-49049112196&partnerID=MN8TOARS KW - Foxp3 KW - regulatory T cells KW - swine ER - TY - JOUR TI - Homologous bone pin conserved in glycerin at 98% and hernicerclagem with a thread of poliglactina 910 in humerus osteosyntesis of domestic pigeons AU - Bolson, Juliano AU - Wallau Schossler, Joao Eduardo AU - Machado, Gustavo AU - Zembrzuski, Fernanda Boligon T2 - Ciencia Rural AB - O objetivo do presente estudo foi avaliar a utilização de um pino intramedular ósseo homólogo, conservado em glicerina a 98%, associado à hemicerclagem com fio poliglactina 910 na osteossíntese umeral transversa de pombos domésticos (Columba livia). Utilizou-se como pino ósseo a parte distal do tibiotarso, conservado com medula óssea. Foram operados 20 pombos domésticos, adultos, não-sexados e clinicamente sadios. Esses animais foram separados ao acaso em cinco grupos, com quatro indivíduos. O úmero foi seccionado cirurgicamente na sua diáfise, de forma transversa, e o pino ósseo foi devidamente implantado no seio do osso. Orifícios foram realizados e por eles procedeu-se hemicerclagem do tipo pontos de Wolff com poliglactina 910. Os animais foram avaliados clinicamente, por meio de radiografias semanais, e histologicamente após os períodos de 15, 30, 60, 90 e 120 dias. A conservação do pino ósseo em glicerina a 98% foi avaliada por meio de exames microbiológicos. Em todos os animais, pode-se notar, clinicamente, excelente adaptação à cirurgia e normalidade do uso do membro já radiologicamente pode-se observar formação de calo ósseo e cicatrização da fratura. Histologicamente pode-se observar que ocorreu formação de calo ósseo 15 dias após a cirurgia e cicatrização com remodelação completa a partir dos 90 dias. Nesse exame, além de leve a moderada reação inflamatória no período inicial de avaliação, nenhum outro evento foi notado nos períodos subseqüentes. Concluiu-se que a técnica e os materiais utilizados se tornam uma opção altamente viável na osteossíntese umeral transversa de pombos domésticos. DA - 2008/// PY - 2008/// DO - 10.1590/s0103-84782008000700019 VL - 38 IS - 7 SP - 1925-1931 UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-54549085568&partnerID=MN8TOARS KW - surgery KW - orthopedics KW - birds KW - bone implant KW - glycerin ER - TY - JOUR TI - The well-behaved dog AU - Alpi, K. M. AU - Sherman, B. L. T2 - Library Journal DA - 2008/// PY - 2008/// VL - 133 IS - 18 SP - 38-39 ER - TY - PAT TI - Immunoglobulin E detection in mammalian species AU - Hammerberg, B. C2 - 2008/// DA - 2008/// PY - 2008/// ER - TY - JOUR TI - Identification of Marek's disease virus genes mutated during serial passage-induced attenuation AU - Spatz, S. J. AU - Gimeno, I. M. AU - Heidari, M. T2 - Vaccine DA - 2008/// PY - 2008/// ER - TY - JOUR TI - Genetic Alterations in K-ras and p53 Cancer Genes in Lung Neoplasms from B6C3F1 Mice Exposed to Cumene AU - Hong, Hue-Hua L. AU - Ton, Thai-Vu T. AU - Kim, Yongbaek AU - Wakamatsu, Nobuko AU - Clayton, Natasha P. AU - Chan, Po-Chuen AU - Sills, Robert C. AU - Lahousse, Stephanie A. T2 - TOXICOLOGIC PATHOLOGY AB - The incidences of alveolar/bronchiolar adenomas and carcinomas in cumene-treated B6C3F1 mice were significantly greater than those of the control animals. We evaluated these lung neoplasms for point mutations in the K-ras and p53 genes that are often mutated in humans. K-ras and p53 mutations were detected by cycle sequencing of PCR-amplified DNA isolated from paraffin-embedded neoplasms. K-ras mutations were detected in 87% of cumene-induced lung neoplasms, and the predominant mutations were exon 1 codon 12 G to T transversions and exon 2 codon 61 A to G transitions. P53 protein expression was detected by immunohistochemistry in 56% of cumene-induced neoplasms, and mutations were detected in 52% of neoplasms. The predominant mutations were exon 5, codon 155 G to A transitions, and codon 133 C to T transitions. No p53 mutations and one of seven (14%) K-ras mutations were detected in spontaneous neoplasms. Cumene-induced lung carcinomas showed loss of heterozygosity (LOH) on chromosome 4 near the p16 gene (13%) and on chromosome 6 near the K-ras gene (12%). No LOH was observed in spontaneous carcinomas or normal lung tissues examined. The pattern of mutations identified in the lung tumors suggests that DNA damage and genomic instability may be contributing factors to the mutation profile and development of lung cancer in mice exposed to cumene. DA - 2008/7// PY - 2008/7// DO - 10.1177/0192623308320280 VL - 36 IS - 5 SP - 720-726 SN - 1533-1601 KW - K-ras KW - p53 KW - cumene KW - lung KW - mouse ER - TY - JOUR TI - Unionicola (Coelaturicola) Edwardsi N. sp (Acari : Unionicolidae) from freshwater mussels in Angola, Africa AU - Vidrine, Malcolm F. AU - Bogan, Arthur E. AU - Bastian-Stanford, Myriam T2 - INTERNATIONAL JOURNAL OF ACAROLOGY AB - Abstract Unionicola (Coelaturicola) edwardsi n. sp. (Acari: Hydrachnidia: Unionicolidae) is described from freshwater mussels, Chambardia moutai (Dartevelle, 1939) (Unionoida: Iridinidae), from the Cunene River Basin, Provincia da Huila, Cape Longo, upper Onquenha River, Angola, Africa. The new species has unique and distinctive male genital field morphology. The species illustrates sexual dimorphism in the chaetotaxy of the first and fourth walking legs. It further extends our knowledge of host-specificity among the Unionicola. Also, new host and geographic records are reported for Unionicola (Mutelicola) blayi Gledhill and Vidrine, 2002, and U. (Coelaturicola) gledhilli Vidrine et al., 2007b. DA - 2008/9// PY - 2008/9// DO - 10.1080/01647950808684542 VL - 34 IS - 3 SP - 277-280 SN - 1945-3892 KW - Acari KW - Hydrachnidia KW - Unionicolidae KW - Unionicola KW - Coelaturicola KW - Mutelicola KW - freshwater mussels KW - Angola ER - TY - JOUR TI - Hemolymph as a nonlethal and minimally invasive source for DNA for molecular systematic studies of freshwater mussels AU - Raley, M. E. AU - Levine, J. F. AU - Bogan, A. E. T2 - Tentacle. The Newsletter of the IUCN/SSC Mollusc Specialist Group DA - 2008/// PY - 2008/// VL - 14 SP - 33–34 ER - TY - JOUR TI - Freshwater bivalve (Unioniformes) diversity, systematics, and evolution: status and future directions AU - Bogan, Arthur E. AU - Roe, Kevin T2 - JOURNAL OF THE NORTH AMERICAN BENTHOLOGICAL SOCIETY AB - Freshwater bivalves of the order Unioniformes represent the largest bivalve radiation in freshwater. The unioniform radiation is unique in the class Bivalvia because it has an obligate parasitic larval stage on the gills or fins of fish; it is divided into 6 families, 181 genera, and ∼800 species. These families are distributed across 6 of the 7 continents and represent the most endangered group of freshwater animals alive today. North American unioniform bivalves have been the subject of study and illustration since Martin Lister, 1686, and over the past 320 y, significant gains have been made in our understanding of the evolutionary history and systematics of these animals. Here, the current state of unioniform systematics and evolution is summarized, and suggestions for future research themes are proposed. Advancement in the areas of systematics and evolutionary relationships within the Unioniformes will require a resurgence of survey work and reevaluation of all taxa, especially outside of North America and Western Europe. This work will require collection of animals for shell morphology, comparative anatomy, and molecular analyses. Along with reexamination of described taxa, a renewed emphasis on the natural history, host-fish relationships, ecology, and physiology of these animals is needed. Traditional conchological and anatomical characters should be reevaluated, new character suites should be added, and new morphometric methods should be applied. The fossil record of freshwater bivalves should be carefully reviewed, and phylogenetic hypotheses including fossil taxa must be developed. We will have to expand our set of molecular tools to include or develop additional markers, such as single-copy nuclear genes and microsatellites. Examination of double uniparental inheritance of mitochondrial deoxyribonucleic acid (DNA) is providing new insights into the evolution of this order. Mitochondrial gene order differs among genera but is still to be explored. Expansion of our understanding of the evolutionary relationships and history of unioniform bivalves will provide a solid foundation to study the zoogeography of these rather sessile, obligate freshwater organisms. The unique natural history of unioniform bivalves provides a fertile area for testing and developing evolutionary theories, and, as our understanding of the systematics of these animals improves, a better understanding of the evolution of this expansive radiation in freshwater will develop. DA - 2008/6// PY - 2008/6// DO - 10.1899/07-069.1 VL - 27 IS - 2 SP - 349-369 SN - 0887-3593 KW - unioniformes KW - mussels KW - bivalves KW - macroinvertebrates KW - benthic KW - clams ER - TY - JOUR TI - Marek's Disease Virus Infection in the Eye: Chronological Study of the Lesions, Virus Replication, and Vaccine-Induced Protection AU - Pandiri, Arun K. R. AU - Cortes, Aneg L. AU - Lee, Lucy F. AU - Gimeno, I. M. T2 - AVIAN DISEASES AB - Marek's disease virus (MDV) infection in the eye was studied chronologically after inoculating 1-day-old chickens with a very virulent MDV strain, Md5. The ocular lesions could be classified as early lesions (6-11 days postinoculation [dpi]) and late lesions (26 and 56 dpi), based upon the location and severity of the lesions. The early lesions involved iris, ciliary body, and choroid layer, and were characterized by endothelial cell hypertrophy, vasculitis, and infiltration of lymphocytes (mainly CD8+), plasma cells, macrophages, and heterophils. Expression of early MDV-antigen pp38 in the cells infiltrating choroid layer was detected as early as 11 dpi. Late lesions consisted of severe lymphohistiocytic uveitis, keratitis, pectenitis, vitreitis, retinitis, and segmental to diffuse retinal necrosis. Cell infiltration included macrophages, granulocytes, plasma cells, and both CD4+ and CD8+ cells of various sizes. Expression of early MDV-antigen pp38 was readily found within the retina, uveal tract, and corneal epithelium. No expression of late-antigen gB or oncoprotein meq was detected in any of the eyes examined. A second experiment was conducted to study the effect of vaccination on the development of ocular lesions. Both HVT and CVI988 were able to protect against the development of early ocular lesions in chickens infected with very virulent plus strain MDV 648A. However, only CVI988 conferred complete protection against the development of late ocular lesions. HVT conferred partial protection, as it reduced the frequency and severity of the late ocular lesions. These results enhance our understanding of the nature and pattern of MDV infection in the eye. DA - 2008/12// PY - 2008/12// DO - 10.1637/8284-031308-Reg.1 VL - 52 IS - 4 SP - 572-580 SN - 1938-4351 KW - Marek's diseases KW - eye KW - retina KW - pathogenesis ER - TY - JOUR TI - Global diversity of freshwater mussels (Mollusca, Bivalvia) in freshwater AU - Bogan, Arthur E. T2 - HYDROBIOLOGIA DA - 2008/1// PY - 2008/1// DO - 10.1007/s10750-007-9011-7 VL - 595 SP - 139-147 SN - 1573-5117 KW - bivalve KW - etheriidae KW - extinction KW - freshwater mussel KW - hyriidae KW - iridinidae KW - margaritiferidae KW - mycetopodidae KW - unionidae KW - unioniformes ER - TY - JOUR TI - MORTALITIES ASSOCIATED WITH SEPSIS, PARASITISM, AND DISSEMINATED ROUND CELL NEOPLASIA IN YELLOW-LIPPED SEA KRAITS (LATICAUDA COLUBRINA) AU - Chinnadurai, Sathya K. AU - Brown, Danielle L. AU - Van Wettere, Arnaud AU - Tuttle, Allison D. AU - Fatzinger, Michael H. AU - Linder, Keith E. AU - Harms, Craig A. T2 - JOURNAL OF ZOO AND WILDLIFE MEDICINE AB - This case series describes multiple mortalities associated with sepsis, neoplasia, and endoparasitism in yellow-lipped sea kraits (Laticauda colubrina) at an exhibit aquarium. Over a 2-yr period, the facility kept 42 L colubrina, of which 38 died and 19 were suitable for necropsy and histopathology. The common clinical syndrome seen in these animals consisted of partial to compete anorexia, increased time spent "hauled-out" on land, intermittent regurgitation, chronic lethargy, and weight loss. Few animals died without premonitory signs. Nutritional support and treatment for presumptive parasitism and sepsis were unsuccessful. The mortality seen in this collection of sea kraits could be placed into three groups; one group of animals (n=9) died of sepsis secondary to necrotizing enteritis or pneumonia; one group (n=6) remained apparently healthy for over 1 yr and then died with multifocal granulomas and sepsis; and the last group (n=3) died as a result of multicentric lymphoid neoplasia with secondary sepsis. The unifying factor in the majority of these cases is the presence of septicemia as the proximate cause of death. Based on the clinical picture, it is presumed that an immunosuppressive event, such as transport, captivity stress, or possible concurrent viral infection, resulted in a septic event and death. DA - 2008/12// PY - 2008/12// DO - 10.1638/2008-0018.1 VL - 39 IS - 4 SP - 626-630 SN - 1937-2825 KW - Laticauda colubrina KW - neoplasia KW - parasite KW - sepsis KW - yellow-tipped sea krait ER - TY - JOUR TI - IMHA: Diagosing and treating a complex disease AU - Shaw, N. AU - Harrell, K. T2 - Veterinary Medicine DA - 2008/// PY - 2008/// VL - 103 IS - 12 SP - 660-671 ER - TY - JOUR TI - CD4(+)CD25(+) regulatory T cells are infected and activated during acute FIV infection AU - Mexas, Angela M. AU - Fogle, Jonathan E. AU - Tompkins, Wayne A. AU - Tompkins, Mary B. T2 - VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY AB - HIV-induced AIDS may be mediated by the activation of immunosuppressive CD4+CD25+ T regulatory cells (Treg cells). Treg cells have been shown to regulate CD4+ and CD8+ immune responses to HIV and FIV antigens in vitro. We tested the hypothesis that Treg cells become infected and activated during the acute infection with FIV leading to the suppression of CD4+ T helper cell responses. Cats were experimentally infected with FIV-NCSU1 and blood and lymph node cells were collected at weekly intervals following inoculation. Real-time RT-PCR was used to determine plasma viremia and the relative expression of FIV, FoxP3, TGF-β, and GAPDH mRNA copies in CD4+CD25+ and CD4+CD25− T cell subsets. Flow cytometry was used to assess the absolute numbers of each cell type and the expression of surface TGF-β and intracellular FoxP3 in CD4+CD25+ and CD4+CD25− T cells at each time-point. Treg suppression of IL-2 production in CD4+ T helper cells was assessed by ELISPOT assays. Our results showed that peak viremia occurred at 2 weeks post infection and correlated with maximal infectivity in CD4+CD25+ T cell populations. FIV-gag-mRNA levels were higher in CD4+CD25+ T cells than CD4+CD25− T cells throughout the acute phase of infection. Induction of FoxP3 and TGF-β indicated activation of Treg cells during the acute stage infection, which was confirmed by Treg cell suppression of IL-2 production by CD4+ Th cells in an ELISPOT assay. Our findings support the hypothesis that early activation of Treg immunosuppressor function may limit an effective anti-FIV response, contributing to the establishment of chronic infection and the immunodeficiency caused by this virus. DA - 2008/12/15/ PY - 2008/12/15/ DO - 10.1016/j.vetimm.2008.08.003 VL - 126 IS - 3-4 SP - 263-272 SN - 1873-2534 KW - FIV KW - HIV KW - Regulatory T cells KW - Acute infection ER - TY - JOUR TI - Broiler embryo bone development is influenced by incubator temperature, oxygen concentration and eggshell conductance at the plateau stage in oxygen consumption AU - Oviedo-Rondon, E. O. AU - Small, J. AU - Wineland, M. J. AU - Christensen, V. L. AU - Mozdziak, P. S. AU - Koci, M. D. AU - Funderburk, S. V. L. AU - Ort, D. T. AU - Mann, K. M. T2 - BRITISH POULTRY SCIENCE AB - 1. Four experiments were conducted to evaluate the effects of temperature (TEM) and oxygen (O2) concentrations during the last 4 d of incubation on bone development. Fertile eggs from two strains were obtained that either exhibited Low or High eggshell conductance (G). 2. Four experimental cabinets provided either four TEM (36, 37, 38 or 39°C) or four O2 concentrations (17, 19, 21 or 23% O2). Data were analysed as a 2 × 2 factorial design. In the fourth experiment, two temperatures (36 and 39°C), two O2 concentrations (17 and 23%) and the same Low and High G strains were evaluated in a 2 × 2 × 2 factorial design. 3. Body weights (BW) and residual yolks were obtained, both legs were dissected. Femur, tibia and shank weights, length and thickness were recorded. Relative asymmetry (RA) of each leg section was calculated. 4. The results indicated that elevated TEM during incubation increased RA between the two legs, mainly in the Low G strain. Chickens at the lowest O2 concentrations had lighter and shorter tibias, lighter shanks, and increased RA of femur length compared to chickens in the 23% O2. In the fourth experiment no interactions were observed between O2 and TEM. High TEM depressed BW of Low G broilers, but no significant effect of treatments was observed on BW of High G broilers. Nevertheless, the high TEM or low O2 independently caused reduced femur and tibia weights and length, shank length and thickness, and both low O2 and high TEM together increased RA in shank weight. 5. These results suggest that late incubation conditions affect long bone development in broilers. DA - 2008/// PY - 2008/// DO - 10.1080/00071660802433149 VL - 49 IS - 6 SP - 666-676 SN - 1466-1799 ER - TY - BOOK TI - Veterinary pharmacology and therapeutics AU - Riviere, J. E. AU - Papich, M. DA - 2008/// PY - 2008/// VL - 9th ed. PB - Ames, IA: Blackwell Publishing SN - 0813820618 ER - TY - CHAP TI - Solvents and vehicle effects on the skin AU - Roberts, M. S. AU - Gierden, A. AU - Riviere, J. E. AU - Monteiro-Riviere, N. A. T2 - Dermal absorption and toxicity assessment. (2nd ed.) A2 - Roberts, M. S. A2 - Walters, K. A. PY - 2008/// SP - 433-447 PB - New York: Informa Healthcare SN - 9780849375910 ER - TY - JOUR TI - Evaluation of four DNA extraction methods for the detection of Tritrichomonas foetus in feline stool specimens by polymerase chain reaction AU - Stauffer, Stephen H. AU - Birkenheuer, Adam J. AU - Levy, Michael G. AU - Marr, Henry AU - Gookin, Jody L. T2 - JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION AB - Feces are increasingly valued as practical samples for molecular diagnosis of infectious disease. However, extraction of polymerase chain reaction (PCR) quality DNA from fecal samples can be challenging because of coextraction of PCR inhibitors. Because the type and quantity of PCR inhibitors is influenced by diet, endogenous flora, and concurrent disease, it is unlikely that extraction method performance with human feces can be directly extrapolated to that of domestic cats. In the present study, 4 commercially available DNA extraction methods were examined for their influence on the sensitivity of PCR for the detection of Tritrichomonas foetus in feline stool. DNA was extracted from serially diluted feline-origin T. foetus trophozoites in the absence or presence of feline feces. The ZR Fecal DNA kit was identified as affording the greatest analytical sensitivity and reproducibility and was able to detect >or=10 T. foetus organisms per 100 mg feces in 100% of PCR reactions. Further, the identified extraction method could be completed in the shortest time of all kits tested. DA - 2008/9// PY - 2008/9// DO - 10.1177/104063870802000518 VL - 20 IS - 5 SP - 639-641 SN - 1943-4936 KW - Cats KW - feces KW - polymerase chain reaction KW - protozoa ER - TY - JOUR TI - Proof of concept of a method that assesses the spread of microbial infections with spatially explicit and non-spatially explicit data AU - Rivas, Ariel L. AU - Anderson, Kevin L. AU - Lyman, Roberta AU - Smith, Stephen D. AU - Schwager, Steven J. T2 - INTERNATIONAL JOURNAL OF HEALTH GEOGRAPHICS AB - A method that assesses bacterial spatial dissemination was explored. It measures microbial genotypes (defined by electrophoretic patterns or EP), host, location (farm), interfarm Euclidean distance, and time. Its proof of concept (construct and internal validity) was evaluated using a dataset that included 113 Staphylococcus aureus EPs from 1126 bovine milk isolates collected on 23 farms between 1988 and 2005.Construct validity was assessed by comparing results based on the interfarm Euclidean distance (a spatially explicit measure) and those produced by the (non-spatial) interfarm number of isolates reporting the same EP. The distance associated with EP spread correlated with the interfarm number of isolates/EP (r = .59, P < 0.02). Internal validity was estimated by comparing results obtained with different versions of the same indices. Concordance was observed between: (a) EP distance (estimated microbial dispersal over space) and EP speed (distance/year, r = .72, P < 0.01), and (b) the interfarm number of isolates/EP (when measured on the basis of non-repeated cow testing) and the same measure as expressed by repeated testing of the same animals (r = .87, P < 0.01). Three EPs (2.6% of all EPs) appeared to be super-spreaders: they were found in 26.75% of all isolates. Various indices differentiated local from spatially disseminated infections and, within the local type, infections suspected to be farm-related were distinguished from cow-related ones.Findings supported both construct and internal validity. Because 3 EPs explained 12 times more isolates than expected and at least twice as many isolates as other EPs did, false negative results associated with the remaining EPs (those erroneously identified as lacking spatial dispersal when, in fact, they disseminated spatially), if they occurred, seemed to have negligible effects. Spatial analysis of laboratory data may support disease surveillance systems by generating hypotheses on microbial dispersal ability. DA - 2008/11/18/ PY - 2008/11/18/ DO - 10.1186/1476-072x-7-58 VL - 7 SP - SN - 1476-072X ER - TY - CHAP TI - Percutaneous absorption of complex chemical mixtures AU - Riviere, J. E. T2 - Marzulli and Maibach's dermatotoxicology (7th ed.) PY - 2008/// SP - 63-70 PB - New York: Taylor and Francis SN - 9780849397738 ER - TY - CHAP TI - Animal skin morphology and dermal absorption AU - Monteiro-Riviere, N. A. AU - Baynes, R. E. AU - Riviere, J. E. T2 - Dermal absorption and toxicity assessment A2 - Roberts, M.S. A2 - Walters, K.A. PY - 2008/// ET - 2nd SP - 17–35 PB - Informa Healthcare SN - 0849375916 ER - TY - JOUR TI - Magnetic resonance imaging of live freshwater mussels (Unionidae) AU - Holliman, F. Michael AU - Davis, Denise AU - Bogan, Arthur E. AU - Kwak, Thomas J. AU - Cope, W. Gregory AU - Levine, Jay F. T2 - INVERTEBRATE BIOLOGY AB - Abstract. We examined the soft tissues of live freshwater mussels, Eastern elliptio Elliptio complanata , via magnetic resonance imaging (MRI), acquiring data with a widely available human whole‐body MRI system. Anatomical features depicted in the profile images included the foot, stomach, intestine, anterior and posterior adductor muscles, and pericardial cavity. Noteworthy observations on soft tissue morphology included a concentration of lipids at the most posterior aspect of the foot, the presence of hemolymph‐filled fissures in the posterior adductor muscle, the presence of a relatively large hemolymph‐filled sinus adjacent to the posterior adductor muscle (at the ventral‐anterior aspect), and segmentation of the intestine (a diagnostic description not reported previously in Unionidae). Relatively little is known about the basic biology and ecological physiology of freshwater mussels. Traditional approaches for studying anatomy and tissue processes, and for measuring sub‐lethal physiological stress, are destructive or invasive. Our study, the first to evaluate freshwater mussel soft tissues by MRI, clarifies the body plan of unionid mussels and demonstrates the efficacy of this technology for in vivo evaluation of the structure, function, and integrity of mussel soft tissues. DA - 2008/// PY - 2008/// DO - 10.1111/j.1744-7410.2008.00143.x VL - 127 IS - 4 SP - 396-402 SN - 1744-7410 KW - MRI KW - morphology KW - bivalve KW - soft tissue KW - unionid ER - TY - JOUR TI - Load of Challenge Marek's Disease Virus DNA in Blood as a Criterion for Early Diagnosis of Marek's Disease Tumors AU - Gimeno, Isabel M. AU - Cortes, Aneg L. AU - Silva, R. F. T2 - AVIAN DISEASES AB - Outbreaks of Marek's disease (MD) in vaccinated flocks still occur sporadically and lead to economic losses. Unfortunately, adequate methods to predict MD outbreaks are lacking. In the present study, we have evaluated whether high load of challenge MD virus (MDV) DNA in peripheral blood could aid in the early diagnosis of MD and in monitoring efficacy of vaccines against MD. One experiment was conducted to simulate field conditions by combining various vaccines (turkey herpesvirus [HVT] and HVT MDV serotype 2 [SB1]) and challenge viruses (GA, Md5, and 648A). Vaccine efficacy among our experimental groups ranged from 13.3% to 94.2%. Each chicken was sampled three times during the length of the experiment (3, 5, and 15 wk postchallenge [wpc]), and gross lesions were evaluated in chickens that died and at termination of the experiment. DNA was extracted from whole blood and buffy coats from each sample, and the load of challenge MDV DNA and HVT DNA were quantified by real-time polymerase chain reaction. Chickens that developed MD by the end of the experiment had higher load of challenge MDV DNA (threshold cycle [Ct] glyceraldehyde-3-phosphate dehydrogenase [GAPDH]/Ct glycoprotein B [gB] ratios of 1.0, 1.04, and 1.05 at 3, 5, and 15 wpc, respectively) than those that did not develop MD (Ct GAPDH/Ct gB ratios of 0.7, 0.69, and 0.46 at 3, 5, and 15 wpc, respectively). However, load of HVT DNA in blood was not correlated with the development of tumors (Ct GAPDH/Ct HVT ratios from 0.04 to 0.10 in both groups). Vaccinated groups with >75% protection had statistically significant less challenge DNA virus (Ct GAPDH/Ct gB ratios of 0.76, 0.70, and 0.45 at 3, 5, and 15 wpc, respectively) than less protected groups (Ct GAPDH/Ct gB ratios of 0.92, 0.97, and 0.85 at 3, 5, and 15 wpc, respectively). No differences in the load of HVT DNA could be found between protected and nonprotected groups at any time point of the study (Ct GAPDH/Ct HVT from 0.05 to 0.09 in both groups). Our results showed that load of challenge MDV DNA but not load of HVT DNA in blood can be used as criterion for early diagnosis of MD. DA - 2008/6// PY - 2008/6// DO - 10.1637/8089-081407-Reg.1 VL - 52 IS - 2 SP - 203-208 SN - 0005-2086 KW - Marek's disease KW - vaccination KW - control KW - protection KW - monitoring ER - TY - JOUR TI - Aural Cholesteatoma in Twenty Dogs AU - Hardie, Elizabeth M. AU - Linder, Keith E. AU - Pease, Anthony P. T2 - VETERINARY SURGERY AB - To determine the clinical course in dogs with aural cholesteatoma.Case series.Dogs (n=20) with aural cholesteatoma.Case review (1998-2007).Twenty dogs were identified. Clinical signs other than those of chronic otitis externa included head tilt (6 dogs), unilateral facial palsy (4), pain on opening or inability to open the mouth (4), and ataxia (3). Computed tomography (CT) was performed in 19 dogs, abnormalities included osteoproliferation (13 dogs), lysis of the bulla (12), expansion of the bulla (11), bone lysis in the squamous or petrosal portion of the temporal bone (4) and enlargement of associated lymph nodes (7). Nineteen dogs had total ear canal ablation-lateral bulla osteotomy or ventral bulla osteotomy with the intent to cure; 9 dogs had no further signs of middle ear disease whereas 10 had persistent or recurrent clinical signs. Risk factors for recurrence after surgery were inability to open the mouth or neurologic signs on admission and lysis of any portion of the temporal bone on CT imaging. Dogs admitted with neurologic signs or inability to open the mouth had a median survival of 16 months.Early surgical treatment of aural cholesteatoma may be curative. Recurrence after surgery is associated with advanced disease, typically indicated by inability to open the jaw, neurologic disease, or bone lysis on CT imaging.Presence of aural cholesteatoma may affect the prognosis for successful surgical treatment of middle ear disease. DA - 2008/12// PY - 2008/12// DO - 10.1111/j.1532-950X.2008.00455.x VL - 37 IS - 8 SP - 763-770 SN - 1532-950X ER - TY - JOUR TI - A solvatochromatic approach to quantifying formulation effects on dermal permeability AU - Baynes, R. E. AU - Xia, X-R. AU - Vijay, V. AU - Riviere, J. E. T2 - SAR AND QSAR IN ENVIRONMENTAL RESEARCH AB - Dermal risk assessments are most often concerned with the occupational and environmental exposure to a single chemical and then determining solute permeability through in vitro or in vivo experimentation with various animal models and/or computational approaches. Oftentimes, the skin is exposed to more than one chemical that could potentially modulate dermal permeability of the chemical that could cause adverse health effects. The focus of this article is to demonstrate that these formulation effects on dermal permeability can occur with simple solvent formulations or complex industrial formulations and that these effects can be modeled within the context of a linear solvation energy relationship (LSER). This research demonstrated that formulation-specific strength coefficients (r p a b v) predicted (r 2 = 0.75–0.83) changes in the dermal permeability of phenolic compounds when formulated with commercial metal-working fluid (MWF) formulations or 50% ethanol. Further experimentation demonstrated that chemical-induced changes in skin permeability with 50% ethanol are strongly correlated (r 2 = 0.91) to similar changes in an inert membrane-coated fiber (MCF) array system consisting of three chemically diverse membranes. Changes in specific strength coefficients pertaining to changes in hydrogen donating ability (Δb) and hydrophobicity (Δv) across membrane systems were identified as important quantitative interactions associated with ethanol mixtures. This solvatochromatic approach along with the use of a MCF array system holds promise for predicting dermal permeability of complex chemical formulations in occupational exposures where performance additives can potentially modulate permeability of potential toxicants. DA - 2008/// PY - 2008/// DO - 10.1080/10629360802551026 VL - 19 IS - 7-8 SP - 615-630 SN - 1029-046X KW - mixtures KW - skin KW - membrane-coated fiber KW - solvation energy ER - TY - JOUR TI - Bacterial orchitis and epididymo-orchitis in broiler breeders AU - Monleon, Rafael AU - Martin, Michael P. AU - Barnes, H. John T2 - AVIAN PATHOLOGY AB - Two cases of orchitis and epididymo-orchitis in broiler breeders are described. The first case occurred in a 62-week-old rooster showing signs of breeding inactivity. Grossly, the left testis was abnormally shaped and had multiple yellow foci, prominent blood vessels, and a gelatinous mass on the surface. The right testis and other tissues appeared normal. A pure, heavy growth of Staphylococcus aureus was obtained on bacterial culture, and intralesional Gram-positive cocci were numerous on histopathology. The second case occurred in a 28-week-old rooster that was found dead. Both testes were abnormally shaped, swollen, firm, and had irregular, dark, depressed areas, prominent vessels, and multiple petechial haemorrhages. Epididymides were enlarged and nodular. Other lesions in this rooster included poor nutritional condition, distended cloaca, and urate scalding of the abdominal skin below the vent. Microscopically, there was extensive, severe heterophilic intratubular orchitis and epididymitis with intralesional Gram-negative bacteria. A heavy, pure growth of Escherichia coli was obtained from the testes and epididymides. In both cases, the findings suggested that the most probable route of infection was ascending via the ductus deferens. DA - 2008/// PY - 2008/// DO - 10.1080/03079450802499134 VL - 37 IS - 6 SP - 613-617 SN - 0307-9457 ER - TY - JOUR TI - Pharmacokinetics of carbetocin, a long-acting oxytocin analogue, following intravenous administration in horses AU - Schramme, A. R. AU - Pinto, C. R. F. AU - Davis, J. AU - Whisnant, C. S. AU - Whitacre, M. D. T2 - EQUINE VETERINARY JOURNAL AB - Current therapy protocols to treat persistent post mating endometritis and retained fetal membranes in mares typically include the administration of ecbolic drugs. Evaluation of the pharmacokinetics and tolerability of carbetocin, a long-acting oxytocin analogue, after i.v. administration is required.To determine the pharmacokinetic parameters (principally half-life) of carbetocin in horses.Five mature mares and one gelding received 0.175 mg carbetocin i.v. All animals were monitored periodically throughout the study for elevation in rectal temperature, heart rate, respiratory rate and signs of pain or discomfort. Plasma samples were collected for determination of carbetocin concentrations by radioimmunoassay.Administration of carbetocin was well tolerated by all horses and its half-life was 17.2 min.The half-life of carbetocin is greater than that previously reported for oxytocin (6.8 min).Carbetocin is an attractive alternative to oxytocin therapy in broodmare management. DA - 2008/11// PY - 2008/11// DO - 10.2746/042516408X334343 VL - 40 IS - 7 SP - 658-661 SN - 0425-1644 KW - horse KW - oxytocin KW - pharmacokinetics KW - carbetocin ER - TY - JOUR TI - Impacts of Road Crossings on Fish Movement and Community Structure AU - Vander Pluym, Jenny L. AU - Eggleston, David B. AU - Levine, Jay F. T2 - Journal of Freshwater Ecology AB - ABSTRACT We quantified the impact of four commonly used road crossings (bridge, arch culvert, box culvert, and pipe culvert) on stream fish community structure and movement in the Piedmont region of the Cape Fear River basin of North Carolina, USA during summer 2004. We focused on non-perched crossings, which did not present a physical barrier to fish movement. With the exception of species richness, all response variables (fish population size, species diversity, fish index of biotic integrity, and conditional percentage of movement) did not vary significantly with crossing type, position (upstream and downstream), or month. Streams with arch culverts contained significantly higher species richness than streams with bridges. The general lack of stream fish abundance and diversity responses to road crossings may be due to the insensitivity of stream fish community variables to anthropogenic effects, the insensitivity of fish communities to the presence of crossings, the overall resilience of fish communities, or the shifting baseline theory (fish communities having shifted to a different community prior to sampling). Triple-pass electrofishing results suggest that these road crossings had no detectable effect on fish abundance or diversity. DA - 2008/12// PY - 2008/12// DO - 10.1080/02705060.2008.9664244 VL - 23 IS - 4 SP - 565-574 J2 - Journal of Freshwater Ecology LA - en OP - SN - 0270-5060 2156-6941 UR - http://dx.doi.org/10.1080/02705060.2008.9664244 DB - Crossref ER - TY - JOUR TI - Erythrophagocytic low-grade extranodal T-cell lymphoma in a cat AU - Carter, J. E. AU - Tarigo, J. L. AU - Vernau, W. AU - Cecere, T. E. AU - Hovis, R. L. AU - Suter, S. E. T2 - VETERINARY CLINICAL PATHOLOGY AB - A 13-year-old male castrated domestic shorthair cat was presented to the referring veterinarian with a 2-month history of weight loss and lethargy. Splenomegaly, hepatomegaly, nonregenerative anemia, neutropenia, and hyperbilirubinemia were noted. Results of testing for feline immunodeficiency virus, feline leukemia virus, Toxoplasma gondii, and Mycoplasma sp. were negative. On cytologic examination of aspirates from the enlarged spleen and liver, a population of erythrophagocytic round cells was observed. Splenectomy and a liver biopsy were done which revealed a population of CD3+/CD79a- erythrophagocytic mononuclear round cells localized in the hepatic and splenic sinusoids. T-cell PARR (PCR for antigen receptor gene rearrangements) analysis of bone marrow and spleen demonstrated a single band indicative of a clonal proliferation of T cells. Based on the marked splenomegaly, sinusoidal infiltration, lack of lymphadenopathy, and results of cytology, PARR, and immunophenotyping, a diagnosis of low-grade extranodal T-cell lymphoma was made. The cat was treated with chlorambucil and prednisolone; clinical and laboratory abnormalities resolved and the cat has remained clinically normal for 2.5 years. To our knowledge, this report documents the first case of an erythrophagocytic T-cell lymphoma in a cat. The clinicopathologic findings were suggestive of hepatosplenic T-cell lymphoma, a neoplasm described previously only in humans and dogs. DA - 2008/12// PY - 2008/12// DO - 10.1111/j.1939-165X.2008.00073.x VL - 37 IS - 4 SP - 416-421 SN - 0275-6382 KW - Cat KW - erythrophagia KW - hepatosplenic KW - lymphoma KW - PARR ER - TY - JOUR TI - Pharmacokinetics of Butorphanol and Evaluation of Physiologic and Behavioral Effects after Intravenous and Intramuscular Administration to Neonatal Foals AU - Arguedas, M. G. AU - Hines, M. T. AU - Papich, M. G. AU - Farnsworth, K. D. AU - Sellon, D. C. T2 - JOURNAL OF VETERINARY INTERNAL MEDICINE AB - Background: Despite frequent clinical use, information about the pharmacokinetics (PK), clinical effects, and safety of butorphanol in foals is not available. Objectives: The purpose of this study was to determine the PK of butorphanol in neonatal foals after IV and IM administration; to determine whether administration of butorphanol results in physiologic or behavioral changes in neonatal foals; and to describe adverse effects associated with its use in neonatal foals. Animals: Six healthy mixed breed pony foals between 3 and 12 days of age were used. Methods: In a 3‐way crossover design, foals received butorphanol (IV and IM, at 0.05 mg/kg) and IV saline (control group). Butorphanol concentrations were determined by high‐performance liquid chromatography and analyzed using a noncompartmental PK model. Physiologic data were obtained at specified intervals after drug administration. Pedometers were used to evaluate locomotor activity. Behavioral data were obtained using a 2‐hour real‐time video recording. Results: The terminal half‐life of butorphanol was 2.1 hours and C 0 was 33.2 ± 12.1 ng/mL after IV injection. For IM injection, C max and T max were 20.1 ± 3.5 ng/mL and 5.9 ± 2.1 minutes, respectively. Bioavailability was 66.1 ± 11.9%. There were minimal effects on vital signs. Foals that received butorphanol spent significantly more time nursing than control foals and appeared sedated. Conclusions and Clinical Importance: The disposition of butorphanol in neonatal foals differs from that in adult horses. The main behavioral effects after butorphanol administration to neonatal foals were sedation and increased feeding behavior. DA - 2008/// PY - 2008/// DO - 10.1111/j.1939-1676.2008.0200.x VL - 22 IS - 6 SP - 1417-1426 SN - 1939-1676 KW - Drug disposition KW - Feeding KW - Narcotic agonist-antagonist KW - Pedometer KW - Sedation ER - TY - JOUR TI - Ocular toxicity and distribution of subconjunctival and intravitreal rapamycin in horses AU - Douglas, L. C. AU - Yi, N. Y. AU - Davis, J. L. AU - Salmon, J. H. AU - Gilger, B. C. T2 - JOURNAL OF VETERINARY PHARMACOLOGY AND THERAPEUTICS AB - In vitro photosensitivity of rapamycin (RAPA) and ocular toxicity and distribution of intravitreal and subconjunctival RAPA was evaluated in normal horses. RAPA (2.5 mg, 5 mg, and 10 mg) was placed in 10 mL of PBS and maintained in a water bath at 37 degrees C, kept in the dark or subjected to room light, and sampled for up to 3 months for RAPA levels. Six normal adult horses received either 5 mg (n = 2) or 10 mg (n = 2) of RAPA intravitreally or 10 mg (n = 2) subconjunctivally. Ophthalmic exams and electroretinography (ERG) were performed prior to injection and on days 1, 7, 14, and 21 post-injection. Eyes were enucleated and samples were collected for RAPA concentrations and histopathology. No difference in light vs. dark RAPA concentrations was observed, suggesting a lack of RAPA phototoxicity. No evidence of ocular toxicity was noted on ophthalmic examination or histopathology. RAPA was not detected intraocularly 7 days post-injection in eyes receiving subconjunctival RAPA, but was detected in the vitreous at 21 days post-injection. Drug could be detected in both the aqueous and vitreous humor after intravitreal injection. Further study is needed to determine the efficacy of intravitreal RAPA. DA - 2008/12// PY - 2008/12// DO - 10.1111/j.1365-2885.2008.00986.x VL - 31 IS - 6 SP - 511-516 SN - 1365-2885 ER - TY - JOUR TI - Equine lymphoma: What are the prospects for cellular differentiation, early diagnosis and intervention strategies? AU - Roberts, M. C. T2 - EQUINE VETERINARY EDUCATION AB - Equine Veterinary EducationVolume 20, Issue 9 p. 464-466 Equine lymphoma: What are the prospects for cellular differentiation, early diagnosis and intervention strategies? M. C. Roberts, M. C. Roberts College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27606, USA.Search for more papers by this author M. C. Roberts, M. C. Roberts College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27606, USA.Search for more papers by this author First published: 05 January 2010 https://doi.org/10.2746/095777308X336327Citations: 7AboutPDF ToolsExport 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 No abstract is available for this article.Citing Literature Volume20, Issue9September 2008Pages 464-466 RelatedInformation DA - 2008/9// PY - 2008/9// DO - 10.2746/095777308x336327 VL - 20 IS - 9 SP - 464-466 SN - 2042-3292 ER - TY - JOUR TI - Endothelial cell suppression of peripheral blood mononuclear cell trafficking in vitro during acute exposure to feline immunodeficiency virus AU - Hudson, Lola C. AU - Tompkins, Mary B. AU - Meeker, Rick B. T2 - CELL AND TISSUE RESEARCH AB - Trafficking of peripheral blood mononuclear cells (PBMCs) into the brain is a critical step in the initiation of human immunodeficiency virus (HIV)-associated central nervous system disease. To examine potential factors that control trafficking during the earliest stages of infection, PBMC transmigration across a cultured feline brain endothelial cell (BECs) monolayer was measured after selective exposure of various cell types to feline immunodeficiency virus (FIV). Infection of the PBMCs with FIV increased the trafficking of monocytes and CD4 and CD8 T cells. Additional exposure of the BECs to FIV suppressed mean monocyte, CD4 T cell, and CD8 T cell trafficking. B cell trafficking was unaltered by these changing conditions. Subsequent exposure of astrocytes or microglia to FIV altered transmigration of different PBMC subsets in different ways. Treated microglia compared with treated astrocytes decreased monocyte transmigration, whereas B cell transmigration was increased significantly. When both astrocytes and microglia were exposed to FIV, an increase in CD8 T cell transmigration relative to BECs alone, to BECs plus astrocytes, or to BECs plus microglia was demonstrated. Thus, initial exposure of PBMCs to FIV is sufficient to induce a general increase in trafficking, whereas initial exposure of endothelial cells to FIV tends to down-regulate this effect. Selectivity of trafficking of specific PBMC subsets is apparent only after exposure of cells of the central nervous system to FIV in co-culture with the endothelium. DA - 2008/10// PY - 2008/10// DO - 10.1007/s00441-008-0623-7 VL - 334 IS - 1 SP - 55-65 SN - 0302-766X KW - HIV KW - astrocytes KW - microglia KW - monocyte KW - T cell KW - feline ER - TY - JOUR TI - Disseminated large granular lymphoma in a horse AU - Sheats, M. K. AU - Wetter, A. J. N. J. AU - Snyder, L. A. AU - Jones, S. L. T2 - EQUINE VETERINARY EDUCATION AB - Summary A 21‐year‐old pony gelding presented for a 5 week history of diarrhoea, inappetance, progressive weight loss and lethargy. Differential diagnoses for chronic diarrhoea and weight loss in horses include: chronic salmonellosis, sand enteropathy, enterolith, parasitism (strongylosis, cyathostomiasis), NSAID induced ulcerative colitis, inflammatory bowel disease (granulomatous, lymphocytic‐plasmacytic or eosinophilic enterocolitis), gastrointestinal neoplasia (lymphosarcoma, squamous cell carcinoma), antibiotic associated clostridial overgrowth, altered diet or bacterial fermentation, peritonitis, Strongylus vulgaris induced arteriopathy (now quite rare) and abdominal mass or abscess. In this gelding, ante mortem diagnosis of CD3 + intestinal large granular lymphoma was made via cytology of abdominal fluid and immunohistochemistry of a rectal muscle biopsy. This report details the clinical, cytological and immunophenotypic findings of a case of large granular lymphoma in a horse. DA - 2008/9// PY - 2008/9// DO - 10.2746/095777308X343860 VL - 20 IS - 9 SP - 459-463 SN - 2042-3292 KW - horse KW - lymphoma KW - weight loss KW - diarrhoea KW - T lymphocytes ER - TY - JOUR TI - Acute toxicity and tissue distributions of malathion in Ambystoma tigrinum AU - Henson-Ramsey, H. AU - Kennedy-Stoskopf, S. AU - Levine, J. F. AU - Taylor, S. K. AU - Shea, D. AU - Stoskopf, M. K. T2 - ARCHIVES OF ENVIRONMENTAL CONTAMINATION AND TOXICOLOGY DA - 2008/10// PY - 2008/10// DO - 10.1007/s00244-007-9091-4 VL - 55 IS - 3 SP - 481-487 SN - 1432-0703 ER - TY - JOUR TI - Aberrant Innate Immune Responses in TLR-ligand Activated HLA-B27 Transgenic Rat Cells AU - Qian, Bi-Feng AU - Tonkonogy, Susan L. AU - Sartor, R. Balfour T2 - INFLAMMATORY BOWEL DISEASES AB - Commensal enteric microbiota initiate and perpetuate immune-mediated colitis in HLA-B27 transgenic (TG) rats but not wildtype (non-TG) littermates. However, the role of the innate immune response to bacterial components has not been established.We examined responses induced by bacterial adjuvants through Toll-like receptor (TLR) and NOD2 signaling in T-cell-depleted splenocytes from HLA-B27 TG rats versus non-TG controls.We found that various bacterial adjuvants induced TNF production by cells obtained from specific pathogen-free (SPF) and germ-free (GF, sterile) TG and non-TG rats. Peptidoglycan-polysaccharide (PG-PS), lipopolysaccharide (LPS), and CpG DNA motifs stimulated higher levels of TNF production by SPF TG rat spleen cells compared to non-TG cells. CD11b/c cell depletion eliminated PG-PS and LPS-induced TNF and dramatically reduced CpG-stimulated TNF production. Both SPF and GF TG rat spleens contain more cells that express high levels of CD11b/c and show enhanced mRNA expression of TLR-2 and TLR-4 compared to non-TG rat spleens. In contrast, constitutive and bacterial-induced IL-10 production was markedly lower in TG cells compared to non-TG cells of rats from the same SPF or GF housing conditions. Notably, the ratio of TNF to IL-10 produced after TLR ligand activation was significantly higher in TG than non-TG cells.HLA-B27 TG rats have an aberrant cell composition, altered functional TLR expression, and an intrinsic defect in IL-10 production in response to TLR ligands, which may result in exaggerated proinflammatory responses to commensal enteric bacteria and uncontrolled inflammation in this colitis model. DA - 2008/10// PY - 2008/10// DO - 10.1002/ibd.20502 VL - 14 IS - 10 SP - 1358-1365 SN - 1536-4844 KW - HLA-B27 transgenic rats KW - bacterial adjuvants KW - Toll-like receptors KW - innate immune responses KW - splenocytes ER - TY - JOUR TI - Valve Gape Response to Turbidity in Two Freshwater Bivalves (Corbicula flumineaandLampsilis radiata) AU - Bucci, John P. AU - Showers, William J. AU - Levine, Jay F. AU - Usry, Brian T2 - Journal of Freshwater Ecology AB - ABSTRACT In a laboratory study with a simulated storm event, we found a significant difference (p<0.05) in valve gape response to the turbidity between Corbicula fluminea and Lampsilis radiata. Valves of C. fluminea opened more intensively during the peak turbidity period and closed significantly more during a following chronic turbidity period. L. radiata exhibited little change in valve gape response with change in turbidity. DA - 2008/9// PY - 2008/9// DO - 10.1080/02705060.2008.9664229 VL - 23 IS - 3 SP - 479-483 J2 - Journal of Freshwater Ecology LA - en OP - SN - 0270-5060 2156-6941 UR - http://dx.doi.org/10.1080/02705060.2008.9664229 DB - Crossref ER - TY - JOUR TI - Comparison of the chloride channel activator lubiprostone and the oral laxative Polyethylene Glycol 3350 on mucosal barrier repair in ischemic-injured porcine intestine AU - Moeser, Adam J. AU - Nighot, Prashant K. AU - Roerig, Birgit AU - Ueno, Ryuji AU - Blikslager, Anthony T. T2 - WORLD JOURNAL OF GASTROENTEROLOGY AB - To investigate the effects of lubiprostone and Polyethylene Glycol 3350 (PEG) on mucosal barrier repair in ischemic-injured porcine intestine.Ileum from 6 piglets (approximately 15 kg body weight) was subjected to ischemic conditions by occluding the local mesenteric circulation for 45 min in vivo. Ileal tissues from each pig were then harvested and mounted in Ussing chambers and bathed in oxygenated Ringer's solution in vitro. Intestinal barrier function was assessed by measuring transepithelial electrical resistance (TER) and mucosal-to-serosal fluxes of (3)H-mannitol and (14)C-inulin. Statistical analyses of data collected over a 120-min time course included 2-way ANOVA for the effects of time and treatment on indices of barrier function.Application of 1 micromol/L lubiprostone to the mucosal surface of ischemic-injured ileum in vitro induced significant elevations in TER compared to non-treated tissue. Lubiprostone also reduced mucosal-to-serosal fluxes of (3)H-mannitol and (14)C-inulin. Alternatively, application of a polyethylene laxative (PEG, 20 mmol/L) to the mucosal surface of ischemic tissues significantly increased flux of (3)H-mannitol and (14)C-inulin.This experiment demonstrates that lubiprostone stimulates recovery of barrier function in ischemic intestinal tissues whereas the PEG laxative had deleterious effects on mucosal repair. These results suggest that, unlike osmotic laxatives, lubiprostone stimulates repair of the injured intestinal barrier. DA - 2008/10/21/ PY - 2008/10/21/ DO - 10.3748/wjg.14.6012 VL - 14 IS - 39 SP - 6012-6017 SN - 2219-2840 KW - Intestinal ischemia KW - Barrier function KW - Permeability KW - Laxative KW - Polyethylene Glycol ER - TY - JOUR TI - Ocular distribution and toxicity of intravitreal injection of triamcinolone acetonide in normal equine eyes AU - Yi, N. Y. AU - Davis, J. L. AU - Salmon, J. H. AU - Gilger, B. C. T2 - VETERINARY OPHTHALMOLOGY AB - Abstract Objective To determine ocular distribution and toxicity of a single injection of intravitreal triamcinolone acetonide (TA) in normal horses. Animals studied Six adult horses, donated to North Carolina State University. Procedures Six horses were injected intravitreally with either 10, 20, or 40 mg ( n = 2 each) of TA. The opposite eye of each horse was injected with balanced salt solution (BSS). Ocular toxicity was assessed by biomicroscopy, tonometry, indirect ophthalmoscopy, and electroretinogram. Aqueous humor (AH), vitreous humor (VH), and plasma samples were collected. Horses were euthanized 7 or 21 days after injection and eyes enucleated for histopathology. TA concentrations in AH, VH, and plasma were measured by HPLC. Results Three control eyes and one TA eye developed inflammation after injection or collection of AH. Positive bacterial cultures ( Corynebacterium spp., Staphylococcus spp., and Streptococcus spp.) were obtained from three of these eyes. Other than transient corneal edema in TA injected eyes, which resolved by 7 days after injection, no other changes were observed. TA crystals were visible within the vitreous body. No evidence of TA toxic effect was noted on histopathology. TA was detected in all AH and VH samples from treated eyes following injection. Drug was not detected in the plasma. Conclusions There was no evidence of overt toxicity from intravitreal TA in normal horses and a single intravitreal injection resulted in TA ocular levels for 21 days. However, the risk for bacterial infections with intravitreal injection or anterior chamber aspirations in horses is high. Use of topical and systemic antibiotics after injection is recommended. DA - 2008/// PY - 2008/// DO - 10.1111/j.1463-5224.2008.00636.x VL - 11 SP - 15-19 SN - 1463-5224 KW - horses KW - intravitreal KW - pharmacokinetics KW - toxicity KW - triamcinolone acetonide ER - TY - JOUR TI - Mice lacking the Na(+)/H(+) exchanger 2 have impaired recovery of intestinal barrier function AU - Moeser, Adam J. AU - Nighot, Prashant K. AU - Ryan, Kathleen A. AU - Simpson, Janet E. AU - Clarke, Lane L. AU - Blikslager, Anthony T. T2 - AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY AB - Ischemic injury induces breakdown of the intestinal barrier. Recent studies in porcine postischemic tissues indicate that inhibition of NHE2 results in enhanced recovery of barrier function in vitro via a process involving interepithelial tight junctions. To further study this process, recovery of barrier function was assessed in wild-type (NHE2 +/+ ) and NHE2 −/− mice in vivo and wild-type mice in vitro. Mice were subjected to complete mesenteric ischemia in vivo, after which barrier function was measured by blood-to-lumen mannitol clearance over a 3-h recovery period or measurement of transepithelial electrical resistance (TER) in Ussing chambers immediately following ischemia. Tissues were assessed for expression of select junctional proteins. Compared with NHE2 +/+ mice, NHE2 −/− mice had greater intestinal permeability during the postischemic recovery process. In contrast to prior porcine studies, pharmacological inhibition of NHE2 in postischemic tissues from wild-type mice also resulted in significant reductions in TER. Mucosa from NHE2 −/− mice displayed a shift of occludin and claudin-1 expression to the Triton-X-soluble membrane fractions and showed disruption of occludin and claudin-1 localization patterns following injury. This was qualitatively and quantitatively recovered in NHE2 +/+ mice compared with NHE2 −/− mice by the end of the 3-h recovery period. Serine phosphorylation of occludin and claudin-1 was downregulated in NHE2 −/− postischemia compared with wild-type mice. These data indicate an important role for NHE2 in recovery of barrier function in mice via a mechanism involving tight junctions. DA - 2008/10// PY - 2008/10// DO - 10.1152/ajpgi.00538.2007 VL - 295 IS - 4 SP - G791-G797 SN - 0193-1857 KW - tight junction KW - intestinal permeability KW - ischemia KW - Na(+)/H(+) exchange ER - TY - JOUR TI - Hematology and plasma chemistry reference intervals for mature laboratory pine voles (Microtus pinetorum) as determined by using the nonparametric rank percentile method AU - Harvey, S. B. AU - Krimer, P. M. AU - Correa, M. T. AU - Hanes, M. A. T2 - Journal of the American Association for Laboratory Animal Science DA - 2008/// PY - 2008/// VL - 47 IS - 4 SP - 35-40 ER - TY - JOUR TI - Epibulbar melanoma in a foal AU - McMullen, Richard J. AU - Clode, Alison B. AU - Pandiri, Arun Kumar R. AU - Malarkey, David E. AU - Michau, Tammy Miller AU - Gilger, Brian C. T2 - VETERINARY OPHTHALMOLOGY AB - A case of epibulbar melanoma in a 6-month-old, gelded, chestnut Hanoverian foal is reported. The location and clinical appearance upon initial presentation led to the tentative diagnosis of staphyloma or a congenital mass of unknown origin. An attempt was made to surgically excise the mass under general anesthesia, but due to its infiltrative nature and intraoperative appearance, most, but not all was removed without compromising the integrity of the globe. Histopathological evaluation revealed a multinodular to packeted, poorly demarcated, unencapsulated, infiltrative exophytic melanocytic neoplasm composed of bundles and nests of plump spindloid to polygonal heavily pigmented epithelioid neoplastic cells interspersed with pigment-laden macrophages within a fine fibrovascular stroma. Upon examination after enucleation, neoplastic cells were found to infiltrate into the lateral cornea, sclera and the choroid. This is a unique case of an epibulbar melanoma with choroidal invasion in a foal. Based on the sudden onset and rapid growth as well as the histological evidence of invasion, well-differentiated features, heavy pigmentation, and no apparent mitoses, this neoplasm was considered to be a low-grade malignant melanoma. At 14 months after excision there is no evidence of recurrence. DA - 2008/// PY - 2008/// DO - 10.1111/j.1463-5224.2008.00637.x VL - 11 SP - 44-50 SN - 1463-5216 KW - congenital KW - epibulbar KW - equine KW - melanoma KW - neoplasia ER - TY - JOUR TI - 5-Hydroxytryptamine (5HT)-induced valvulopathy: Compositional. valvular alterations are associated with 5HT2B receptor and 5HT transporter transcript changes in Sprague-Dawley rats AU - Elangbam, Chandikumar S. AU - Job, Lauren E. AU - Zadrozny, Leah M. AU - Barton, Joanna C. AU - Yoon, Lawrence W. AU - Gates, Lisa D. AU - Slocum, Nikki T2 - EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY AB - Several drugs have been linked to valvulopathy in humans, including therapeutic agents for obesity, Parkinson's disease and migraine. There is increasing evidence that the 5-hydroxytryptamine 2B receptor (5HT2BR) activation and/or increased circulating 5HT (5-hydroxytryptamine) may play a significant role in the pathogenesis of drug-induced valvulopathy. In the present study, we investigated whether 7-day 5HT subcutaneous injections led to structural and compositional abnormalities in conjunction with transcriptomic modulation of 5HT2BR and 5HT transporter (5HTT) genes in the aortic and mitral valves of Sprague-Dawley (SD) rats. Subcutaneous injections of 5HT for 7 days resulted in thickening and compositional alteration of aortic and mitral valves in SD rats. More specifically, valve-leaflets from 5HT-treated rats had greater valve thickness, a higher amount of glycosaminoglycans (GAGs) and a lower amount of collagen. The compositional alteration was associated with up-regulation and down-regulation of 5HT2BR and 5HTT genes, respectively. The present study strongly suggests that the activation of 5HT2BR and inhibition of 5HTT played a significant role in the pathogenesis of 5HT-induced valvulopathy in SD rats. Thus, these findings further highlight the necessity and/or utilization of animal models to screen potential valvular effects of serotonergic compounds. DA - 2008/8// PY - 2008/8// DO - 10.1016/j.etp.2008.03.005 VL - 60 IS - 4-5 SP - 253-262 SN - 0940-2993 KW - valvulopathy KW - 5-hydroxytryptamine 2B receptor KW - 5-hydroxytryptamine transporter KW - anorexigens KW - compositional morphometry KW - quantitative image analysis KW - Sprague-Dawley rats ER - TY - JOUR TI - A Listeria monocytogenes mutant defective in bacteriophage attachment is attenuated in orally inoculated mice and impaired in enterocyte intracellular growth AU - Spears, Patricia A. AU - Suyemoto, M. Mitsu AU - Palermo, Angela M. AU - Horton, John R. AU - Hamrick, Terri S. AU - Havell, Edward A. AU - Orndorff, Paul E. T2 - INFECTION AND IMMUNITY AB - ABSTRACT A Listeria monocytogenes bacteriophage was used to identify a phage-resistant Tn 917 insertion mutant of the mouse-virulent listerial strain F6214-1. The mutant was attenuated when it was inoculated orally into female A/J mice and failed to replicate efficiently in cultured mouse enterocytes. Phage binding studies indicated that the mutant had a cell surface alteration that precluded phage attachment. All phenotypes associated with the mutation could be complemented in trans by a single open reading frame (ORF) that corresponded to the ORF interrupted by the Tn 917 insertion. The complementation effected was, in all cases, at a level indistinguishable from that of the parent. The Tn 917 insertion interrupted a gene that is predicted to encode a group 2 glycosyl transferase (provisionally designated glcV ). A similar glcV gene is present in Listeria welshimeri and Listeria innocua and in some serotypes of L. monocytogenes . We speculate that the loss of the glcV product results in a defective phage receptor and that this alteration coincidentally influences a feature of the normal host-pathogen interaction required for virulence. Interestingly, the glcV lesion, while preventing phage attachment, did not alter the mutant's ability to bind to cultured mouse enterocyte monolayers. Rather, the mutation appeared to alter a subsequent step in intracellular replication measured by a reduction in plaque-forming efficiency and plaque size. In vivo, the mutant was undetectable in the liver and spleen 48 h after oral inoculation. The mutation is significant in part because it is one of the few that produce attenuation when the mutant is delivered orally. DA - 2008/9// PY - 2008/9// DO - 10.1128/IAI.00283-08 VL - 76 IS - 9 SP - 4046-4054 SN - 0019-9567 ER - TY - JOUR TI - Estimating meat withdrawal times in pigs exposed to melamine contaminated feed using a physiologically based pharmacokinetic model AU - Buur, Jennifer L. AU - Baynes, Ronald E. AU - Riviere, Jim E. T2 - REGULATORY TOXICOLOGY AND PHARMACOLOGY AB - Recently melamine was found to have contaminated the feed of multiple food production species leading to concern over the ability to establish an appropriate withdrawal interval and protect the safety of the food supply. To establish an appropriate withdrawal interval, a physiologically based pharmacokinetic (PBPK) model for melamine was developed for rats and extrapolated to pigs. The rat model underpredicted plasma concentrations, but better predicted tissue residues. Correlation values for plasma, kidney, and liver were 0.59, 0.76, and 0.73, respectively. The pig model underpredicted early plasma time points but had greater accuracy at later time points which is relevant to withdrawal times. Correlation (R2) between predicted and observed plasma values was 0.89 with a negative intercept of −0.76. The pig model estimated a withdrawal interval (based on kidney tissue residues) of 19.2 and 20.9 h for single oral exposures of 3.0 and 5.12 mg/kg of melamine, respectively. Chronic oral dosing (3.0 and 5.12 mg/kg twice daily for 7 days) yielded withdrawal intervals of 20 and 21.3 h, respectively. PBPK models, such as this one, provide evidence of the usefulness in species extrapolation over a range of dosing scenarios and can be used to protect the food supply after accidental exposure in the face of little in the target species. DA - 2008/8// PY - 2008/8// DO - 10.1016/j.yrtph.2008.05.003 VL - 51 IS - 3 SP - 324-331 SN - 1096-0295 KW - PBPK KW - melamine KW - rat KW - porcine KW - meat withdrawal time KW - feed contaminants ER - TY - JOUR TI - The use of antifungals AU - Davis, J. L. T2 - Veterinary Technician DA - 2008/// PY - 2008/// VL - 29 IS - 7 SP - 428- ER - TY - JOUR TI - Isolation, detection and characterization of swine hepatitis E virus from herds in Costa Rica AU - Kase, Julie A. AU - Correa, Maria T. AU - Luna, Carlos AU - Sobsey, Mark D. T2 - INTERNATIONAL JOURNAL OF ENVIRONMENTAL HEALTH RESEARCH AB - Although swine HEV isolates from North America, Europe, and Asia have been genetically characterized, little is known about the strains presumed to be circulating in Latin America. In this study, seven commercial swine production sites in Costa Rica were surveyed for HEV. Using RT-PCR, with primers located in ORF2, 19/52 fecal samples produced a product of the expected size following two rounds of amplification. Most positive samples were from swine between the ages of 1.5 and 4 months. This study provides documented evidence for the endemicity of HE infections in swine residing in Central America. Through nucleic acid sequencing, isolates were found to be genetically similar, if not identical, with no amino acid substitutions. By comparison of swine and human HEV strains representing all four genotypes and phylogenetic analysis, our isolates closely resembled the US swine and human and other Genotype III strains, with 85-93% nucleic acid identity. DA - 2008/// PY - 2008/// DO - 10.1080/09603120701498311 VL - 18 IS - 3 SP - 165-176 SN - 1369-1619 KW - hepatitis E virus KW - zoonoses KW - public health KW - disease reservoirs KW - surveillance KW - molecular epidemiology ER - TY - JOUR TI - Serologic survey of woodchuck hepatitis virus in North Carolina woodchucks (Marmota monax) AU - Cullen, J. M. AU - Lindsey-Pegram, D. AU - Cote, P. J. T2 - JOURNAL OF ZOO AND WILDLIFE MEDICINE AB - The prevalence of woodchuck hepatitis virus (WHV) in wild populations of woodchucks is understudied and therefore unclear. Although infection is common in the southeastern region of Pennsylvania and surrounding states, it is virtually absent in New York and New England. Sera were collected from wild woodchucks from Orange County, North Carolina and tested for the presence of markers of current or previous infection with WHV. Of the 24 woodchucks tested, there were three animals (12.5%) with WHV surface antigen as well as antibodies to woodchuck hepatitis core antigen in their serum, indicative of active infection. There were four (17%) animals with antibodies to WHV core antigen but no woodchuck hepatitis surface antigen, indicative of prior infections. The remaining 17 animals had no detectable markers of WHV infection. These data indicate that WHV is present in central North Carolina at rates approaching those seen in endemic areas, such as the mid-Atlantic region of the United States. DA - 2008/6// PY - 2008/6// DO - 10.1638/2007-0119R.1 VL - 39 IS - 2 SP - 263-265 SN - 1937-2825 KW - hepadnavirus KW - woodchuck KW - woodchuck hepatitis virus ER - TY - JOUR TI - Secondary amyloidosis and renal failure in a captive California sea lion (Zalophus californianus) AU - Chinnadurai, Sathya K. AU - Van Wettere, Arnaud AU - Linder, Keith E. AU - Harms, Craig A. AU - DeVoe, Ryan S. T2 - JOURNAL OF ZOO AND WILDLIFE MEDICINE AB - A 16-yr-old, captive-born, female California sea lion (Zalophus californianus) was evaluated for intermittent lethargy, partial anorexia, and polydipsia of 2 wk duration. The animal was immobilized for physical examination. It was in thin body condition, with multifocal mucosal ulcerations over the caudal and ventral tongue. Blood was collected for hematology, serum chemistry, and leptospirosis serology. Serum chemistry revealed severe azotemia, mild hyperglycemia, and severe hyperphosphatemia. The animal went into cardiac arrest during recovery from anesthesia and died. On histopathology, abundant amorphous, finely fibrillar, eosinophilic material was deposited in the kidneys, and smaller amounts of the same material were found in the splenic and pancreatic vessels; these findings are consistent with systemic secondary amyloidosis. The animal also had chronic nephritis, which, coupled with renal amyloidosis, resulted in renal failure and death. Systemic amyloidosis should be considered as an additional differential diagnosis for renal failure in California sea lions. DA - 2008/6// PY - 2008/6// DO - 10.1638/2007-0096R.1 VL - 39 IS - 2 SP - 274-278 SN - 1937-2825 KW - California sea lion KW - renal failure KW - secondary amyloidosis KW - Zalophus californianus ER - TY - JOUR TI - Reduced responsiveness of HLA-B27 transgenic rat cells to TGF-beta and IL-10-mediated regulation of IFN-gamma production AU - Qian, Bi-Feng AU - Tonkonogy, Susan L. AU - Sartor, R. Balfour T2 - INFLAMMATORY BOWEL DISEASES AB - We have reported that commensal luminal bacterial components induce an active in vitro IFN-gamma response in mesenteric lymph node (MLN) and intestinal cells from specific pathogen-free (SPF) HLA-B27 transgenic (TG) rats with chronic colitis but not in cells from non-diseased SPF non-TG, germ-free (GF) non-TG or GF TG rats.The study examined IL-12 stimulation of MLN IFN-gamma responses to luminal bacteria and regulation of these responses by suppressive cytokines.Exogenous IL-12 significantly increased the bacterial lysate-induced IFN-gamma response in SPF TG MLN cells, while bacterial lysate and IL-12 synergistically induced IFN-gamma from low baseline levels in cells obtained from both SPF and GF non-TG rats, and in GF TG cells. TGF-beta fully counteracted the effects of IL-12 and bacterial lysate on non-TG cells by almost completely inhibiting IFN-gamma production. In contrast, TG cells were less responsive to TGF-beta-mediated downregulation with a substantial residual IFN-gamma response to IL-12 plus bacterial lysate. Further experiments showed that CD4+/CD25+ cells had no inhibitory effect on the IFN-gamma production and were not required for TGF-beta-mediated suppression. Addition of exogenous IL-10 also partially inhibited IFN-gamma production by non-TG cells but did not affect TG cells. Conversely, exogenous IL-12 preferentially suppressed bacterial lysate-induced TGF-beta and IL-10 production in TG rat cells.An attenuated response to regulatory signals leads to uncontrolled potentiated induction of effector IFN-gamma responses to commensal bacteria in HLA-B27 TG rats that spontaneously develop chronic intestinal inflammation. DA - 2008/7// PY - 2008/7// DO - 10.1002/ibd.20415 VL - 14 IS - 7 SP - 921-930 SN - 1536-4844 KW - HLA-B27 transgenic rats KW - cytokines KW - defective regulation ER - TY - JOUR TI - Primary intimal aortic angiosarcoma in a dog AU - Ranck, R. S. AU - Linder, K. E. AU - Haber, M. D. AU - Meuten, D. J. T2 - VETERINARY PATHOLOGY AB - A primary intimal aortic angiosarcoma was diagnosed in a 4-year-old, spayed female German Shepherd that presented for complications of thromboembolic disease because of infarcts in multiple organs. On gross examination, aneurysmal dilatation of the aorta was associated with a friable, necrotic mass attached to the endothelial surface, which partially occluded the aortic lumen. On histologic examination, plump neoplastic spindle cells formed a plaque-like mass arising from the intima that merged with a large accumulation of fibrin and necrotic debris, and projected into the lumen. Neoplastic cells invaded periaortic vessels and were seen in some infarct-associated thromboemboli. Tumor cells expressed vimentin and CD31, with infrequent, patchy staining with factor VIII-related antigen; tumor cells were negative for cytokeratin and smooth-muscle actin. Aortic angiosarcoma is a rare malignancy in humans. This is the first description of a primary intimal aortic angiosarcoma in a dog, with immunohistochemical evidence of endothelial origin. DA - 2008/5// PY - 2008/5// DO - 10.1354/vp.45-3-361 VL - 45 IS - 3 SP - 361-364 SN - 0300-9858 KW - aneurysm KW - angiosarcoma KW - aorta KW - canines KW - CD31 KW - immunohistochemistry KW - thromboembolism ER - TY - JOUR TI - Lentivirus-induced immune dysregulation AU - Tompkins, Mary B. AU - Tompkins, Wayne A. T2 - VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY AB - FIV/HIV infections are associated with an early robust humoral and cellular anti-viral immune response followed by a progressive immune suppression that eventually results in AIDS. Several mechanisms responsible for this immune dysfunction have been proposed including cytokine dysregulation, immunologic anergy and apoptosis, and inappropriate activation of immune regulatory cells. Studies on FIV infection provide evidence for all three. Cytokine alterations include decreases in IL2 and IL12 production and increases in IFNgamma and IL10 in FIV(+) cats compared to normal cats. The elevated IL10:IL12 ratio is associated with the inability of FIV(+) cats to mount a successful immune response to secondary pathogens. Additionally, chronic antigenic (FIV) stimulation results in an increase in the percent of activated T cells expressing B7 and CTLA4 co-stimulatory molecules in infected cats. The expression of these molecules is associated with T cells that are undergoing apoptosis in the lymph nodes. As ligation of CTLA4 by B7 transduces a signal for induction of anergy, one can speculate that the activated T cells are capable of T cell-T cell interactions resulting in anergy and apoptosis. The inability of CD4(+) cells from FIV(+) cats to produce IL2 in response to recall antigens and the gradual loss of CD4(+) cell numbers could be due to B7-CTLA4 interactions. The chronic antigenemia may also lead to activation of CD4(+)CD25(+) T regulatory cells. Treg cells from FIV(+) cats are chronically activated and inhibit the mitogen-induced proliferative response of CD4(+)CD25(-) by down-regulating IL2 production. Although Treg cell activation can be antigen-specific, the suppressor function is not, and thus activated Treg cells would suppress responses to secondary pathogens as well as to FIV. Concomitant with the well-known virus-induced immune suppression is a progressive immune hyper-activation. Evidence for immune hyper-activation includes polyclonal B cell responses, gradual replacement of naïve CD4(+) and CD8(+) T cell phenotypes with activation phenotypes (CD62L(-), B7(+), CTLA4(+)), and the chronic activation of CD4(+)CD25(+) Treg cells. Thus lentivirus infections lead to severe immune dysregulation manifested as both chronic immune suppression and chronic immune activation. FIV infection of cats provides a number of advantages over other lentivirus infections as a model to study this immune dysregulation. It is a natural infection that has existed in balance with the cat's immune system for thousands of years. As such, the natural history and pathogenesis provides an excellent model to study the long-term relationships between AIDS lentivirus and host immune system function/dysregulation. DA - 2008/5/15/ PY - 2008/5/15/ DO - 10.1016/j.vetimm.2008.01.011 VL - 123 IS - 1-2 SP - 45-55 SN - 0165-2427 KW - FIV KW - AIDS KW - immune suppression KW - immune regulation ER - TY - JOUR TI - Inhibition of jet fuel aliphatic hydrocarbon induced toxicity in human epidermal keratinocytes AU - Inman, A. O. AU - Monteiro-Riviere, N. A. AU - Riviere, J. E. T2 - JOURNAL OF APPLIED TOXICOLOGY AB - Abstract Jet propellant (JP)‐8, the primary jet fuel used by the U.S. military, consists of hydrocarbon‐rich kerosene base commercial jet fuel (Jet‐A) plus additives DC1‐4A, Stadis 450 and diethylene glycol monomethyl ether. Human epidermal keratinocytes (HEK) were exposed to JP‐8, aliphatic hydrocarbon (HC) fuel S‐8 and aliphatic HC pentadecane (penta), tetradecane (tetra), tridecane (tri) and undecane (un) for 5 min. Additional studies were conducted with signal transduction pathway blockers parthenolide (P; 3.0 µ m ), isohelenin (I; 3.0 µ m ), SB 203580 (SB; 13.3 µ m ), substance P (SP; 3.0 µ m ) and recombinant human IL‐10 (rHIL‐10; 10 ng ml −1 ). In the absence of inhibitors, JP‐8 and to a lesser extent un and S‐8, had the greatest toxic effect on cell viability and inflammation suggesting, as least in vitro , that synthetic S‐8 fuel is less irritating than the currently used JP‐8. Each inhibitor significantly ( P < 0.05) decreased HEK viability. DMSO, the vehicle for P, I and SB, had a minimal effect on viability. Overall, IL‐8 production was suppressed at least 30% after treatment with each inhibitor. Normalizing data relative to control indicate which inhibitors suppress HC‐mediated IL‐8 to control levels. P was the most effective inhibitor of IL‐8 release; IL‐8 was significantly decreased after exposure to un, tri, tetra and penta but significantly increased after JP‐8 exposure compared with controls. Inhibitors were not effective in suppressing IL‐8 release in JP‐8 exposures to control levels. This study shows that inhibiting NF‐ κ B, which appears to play a role in cytokine production in HC‐exposed HEK in vitro , may reduce the inflammatory effect of HC in vivo . Copyright © 2007 John Wiley & Sons, Ltd. DA - 2008/5// PY - 2008/5// DO - 10.1002/jat.1309 VL - 28 IS - 4 SP - 543-553 SN - 1099-1263 KW - jet fuel KW - JP-8 KW - skin toxicity KW - S-8 KW - NF-kappa B KW - parthenolide KW - isohelenin KW - substance P KW - SB 203580 KW - recombinant human IL-10 ER - TY - JOUR TI - Harderian gland neoplasms in captive, wild-caught Beechey ground squirrels (Spermophilus beecheyi) AU - Ranck, R. S. AU - Cullen, J. M. AU - Waggie, K. S. AU - Marion, P. L. T2 - VETERINARY PATHOLOGY AB - Harderian gland neoplasms were identified in 18 aged, adult Beechey ground squirrels ( Spermophilus beecheyi) from the records of 167 wild-caught captive animals that were necropsied. All but one animal had tumors that were classified as carcinomas, with infiltrative growth and frequent metastases. This is the first detailed report of Harderian gland neoplasia in wild Sciuridae, although this neoplasm has been described in other rodent species. Clinically, affected ground squirrels typically were inappetent and presented with weight loss and exophthalmos. The biologic behavior of Harderian gland neoplasia is variable among rodent species; in Beechey ground squirrels there was a high incidence of malignant behavior. Eleven of 17 tumor-bearing animals for which the gender was known were male, and 6 were female. Nine of 16 for which data were available were uninfected, and 7 had evidence of current or prior infection with ground squirrel hepatitis virus. Tumor development occurred in older animals; all but 2 were 5.5 years of age or older. The presence of metastasis was not related to gender or chronic ground squirrel hepatitis virus infection. DA - 2008/5// PY - 2008/5// DO - 10.1354/vp.45-3-388 VL - 45 IS - 3 SP - 388-392 SN - 0300-9858 KW - carcinoma KW - ground squirrel hepatitis virus KW - ground squirrels KW - Harderian gland KW - Sciuridae ER - TY - JOUR TI - Formate acts as a diffusible signal to induce Salmonella invasion AU - Huang, Yanyan AU - Suyemoto, Mitsu AU - Garner, Cherilyn D. AU - Cicconi, Kellie M. AU - Altier, Craig T2 - JOURNAL OF BACTERIOLOGY AB - To infect an animal host, Salmonella enterica serovar Typhimurium must penetrate the intestinal epithelial barrier. This process of invasion requires a type III secretion system encoded within Salmonella pathogenicity island I (SPI1). We found that a mutant with deletions of the acetate kinase and phosphotransacetylase genes (ackA-pta) was deficient in invasion and SPI1 expression but that invasion gene expression was completely restored by supplying medium conditioned by growth of the wild-type strain, suggesting that a signal produced by the wild type, but not by the ackA-pta mutant, was required for invasion. This mutant also excreted 68-fold-less formate into the culture medium, and the addition of sodium formate to cultures restored both the expression of SPI1 and the invasion of cultured epithelial cells by the mutant. The effect of formate was pH dependent, requiring a pH below neutrality, and studies in mice showed that the distal ileum, the preferred site of Salmonella invasion in this species, had the appropriate formate concentration and pH to elicit invasion, while the cecum contained no detectable formate. Furthermore, we found that formate affected the major regulators of SPI1, hilA and hilD, but that the primary routes of formate metabolism played no role in its activity as a signal. DA - 2008/6// PY - 2008/6// DO - 10.1128/JB.00205-08 VL - 190 IS - 12 SP - 4233-4241 SN - 1098-5530 ER - TY - JOUR TI - Modulation of pulmonary inflammatory responses and antimicrobial defenses in mice exposed to diesel exhaust AU - Gowdy, Kymberly AU - Krantz, Quentin T. AU - Daniels, Mary AU - Linak, William P. AU - Jaspers, Ilona AU - Gilmour, M. Ian T2 - TOXICOLOGY AND APPLIED PHARMACOLOGY AB - Diesel exhaust (DE) is a major component of urban air pollution and has been shown to increase the severity of infectious and allergic lung disease. The purpose of this study was to evaluate the effects of DE exposure on pulmonary inflammation, mediator production and antimicrobial defenses in an exposure model that had previously been shown to increase susceptibility to influenza. BALB/c mice were exposed to filtered air, or to DE diluted to yield 0.5 or 2 mg/m3 of diesel exhaust particles (DEP) for 4 h per day for 1 or 5 days. Immediately and 18 h after one or five diesel exposures mice were euthanized to assess both immediate and delayed effects. DE exposure for 5 days at either concentration caused higher neutrophil numbers and lesion scoring compared to air controls. Intracellular adhesion molecule-1 (ICAM-1), which recruits inflammatory cells and is an entry site for rhinoviruses was increased immediately after 1 or 5 days of DE exposure. Several inflammatory and immune cytokines (TNF-α, MIP-2, IL-6, IFN-γ, and IL-13) were also upregulated at various time points and concentrations. In contrast, clara cell secretory protein (CCSP), surfactant protein A (SP-A), and surfactant protein D (SP-D) which are important host defense molecules, were significantly decreased at both the message and protein level with DE exposure. We conclude that exposure to moderate and high occupational levels of DE caused an increase in lung injury and inflammation, and a decrease in host defense molecules, which could result in increased susceptibility to respiratory pathogens. DA - 2008/6/15/ PY - 2008/6/15/ DO - 10.1016/j.taap.2008.01.040 VL - 229 IS - 3 SP - 310-319 SN - 1096-0333 KW - diesel exhaust KW - inflammation KW - lung KW - surfactant proteins KW - clara cell KW - mice ER - TY - JOUR TI - Evaluation of wraps covering the distal aspect of pelvic limbs for prevention of bacterial strike-through in an ex vivo canine model AU - Vince, Kent J. AU - Lascelles, B. Duncan X. AU - Mathews, Kyle G. AU - Altier, Craig AU - Roe, Simon C. T2 - VETERINARY SURGERY AB - To determine differences in bacterial strike-through for materials commonly used to cover the distal aspect of the pelvic limb during operative site preparation.Randomized block design; ex vivo model.Canine cadaveric pelvic limbs (n=40).Pelvic limbs (n=40) were randomly assigned to 4 treatment groups: Group 1=Vetrap+sterile Coban; Group 2=latex glove+Vetrap+sterile Coban; Group 3=latex glove+Vetrap+sterile Coban+sterile latex glove+sterile Coban; and Group 4=latex glove+Vetrap+sterile disposable drape+sterile Coban. Limbs were contaminated with a standardized bacterial solution and routinely prepared using the assigned distal leg wrap. Bandages were fluid challenged with a saline (0.9% NaCl) solution-soaked laparotomy sponge for 30 seconds. The wrap surface was sampled for microbial culture before surgical preparation, immediately after, and 60 minutes after applying a sterile leg wrap.Bacterial growth occurred in all Group 1 cultures, 90% of Group 2 cultures, and none of the Group 3 and 4 cultures, 60 minutes after applying the sterile wrap.A distal leg wrap of Vetrap+sterile Coban is not effective in preventing bacterial strike-through.If similar results occur in the live animal, then a sterile impermeable barrier must be incorporated into the distal leg wrap to prevent bacterial strike-through. DA - 2008/6// PY - 2008/6// DO - 10.1111/j.1532-950X.2008.00395.x VL - 37 IS - 4 SP - 406-411 SN - 0161-3499 ER - TY - JOUR TI - Attenuation of ischaemic injury in the equine jejunum by administration of systemic lidocaine AU - Cook, V. L. AU - Shults, J. J. AU - McDowell, M. AU - Campbell, N. B. AU - Davis, J. L. AU - Blikslager, A. T. T2 - Equine Veterinary Journal DA - 2008/// PY - 2008/// DO - 10.2746/04251640SX293574 VL - 40 IS - 4 SP - 353-357 ER - TY - JOUR TI - Marek's disease vaccines: A solution for today but a worry for tomorrow? AU - Gimeno, Isabel M. T2 - VACCINE AB - Marek's disease (MD) is a lymphoproliferative disease of chickens that, in the absence of control measures, is capable of causing devastating losses in commercial poultry flocks. MD has been successfully controlled by vaccination since 1968. However, vaccine efficacy has decreased concomitantly with the increase in virulence of Marek's disease virus (MDV). The constant evolution of MDV has forced the development of new vaccines or vaccine strategies that control the more virulent emergent strains. However, this race between the introduction of new vaccines and the evolution of MDV represents a major threat for the poultry industry. In addition to vaccination, other factors might have contributed to the evolution of MDV (intensive methods of chicken production, early exposure of the chickens to MDV and administration of vaccines at very low doses). From all the possible factors influencing MDV evolution, the effect of vaccination has received the greatest attention. MD vaccines protect with great efficacy against the development of the disease but they do not prevent infection or transmission. Sterilizing immunity could be a solution to stop the evolution of the virus but it has been proven to be extremely difficult, if at all possible, to obtain with MDV or with other herpesviruses. Other solutions to improve vaccine-induced protection are discussed in this paper. DA - 2008/7/18/ PY - 2008/7/18/ DO - 10.1016/j.vaccine.2008.04.009 VL - 26 SP - C31-C41 SN - 1873-2518 KW - Marek's disease KW - virulence KW - vaccines ER - TY - JOUR TI - Effects of incubator temperature and oxygen concentration during the plateau stage of oxygen consumption on turkey embryo long bone development AU - Oviedo-Rondon, E. O. AU - Small, J. AU - Wineland, M. J. AU - Christensen, V. L. AU - Grimes, J. L. AU - Funderburk, S. V. L. AU - Ort, D. T. AU - Mann, K. M. T2 - POULTRY SCIENCE AB - Temperature (TEM) and O2 concentrations during the plateau stage of oxygen consumption are known to affect yolk utilization, tissue development, and thyroid metabolism in turkey embryos. Three experiments were conducted to evaluate these incubation effects on long bone development. Fertile eggs of Nicholas turkeys were used. In each trial, standard incubation conditions were used to 24 d, when the eggs containing viable embryos were randomly divided into 4 groups. Four experimental cabinets provided 4 TEM (36, 37, 38, or 39°C) or 4 O2 concentrations (17, 19, 21, or 23% O2). In the third experiment, 2 temperatures (36 and 39°C) and 2 O2 concentrations (17 and 23%) were evaluated in a 2 × 2 factorial design. Body and residual yolk weights were obtained. Both legs were dissected, and shanks, femur, and tibia weights, length, and thickness were recorded. Relative asymmetry of each leg section was calculated. Chondrocyte density was evaluated in slides stained with hematoxylin and eosin. Immunofluorescence was used to evaluate the presence of collagen type X and transforming growth factor β. Hot TEM caused reduction of tibia weights and increase of shank weight when compared with cool TEM. The lengths of femur, tibia, and shanks were reduced by 39°C. The relative asymmetry of leg weights were increased at 38 and 39°C. Poult body and part weights were not affected by O2 concentrations, but poults on 23% O2 had bigger shanks and heavier tibias than the ones on 17% O2. High TEM depressed the fluorescence of collagen type X and transforming growth factor β. The O2 concentrations did not consistently affect the immunofluorescence of these proteins. The chondrocyte density was affected by TEM and O2 in resting and hypertrophic zones. In the third experiment, high TEM depressed BW, leg muscle weights, and shank length. Low O2 reduced tibia and shanks as a proportion of the whole body. We concluded that incubation conditions affect long bone development in turkeys. DA - 2008/8// PY - 2008/8// DO - 10.3382/ps.2007-00470 VL - 87 IS - 8 SP - 1484-1492 SN - 1525-3171 KW - turkey KW - incubation KW - leg health KW - bone development ER - TY - JOUR TI - Correction of multiple striated muscles in murine Pompe diseasethrough adeno-associated virus-mediated gene therapy AU - Sun, Baodong AU - Young, Sarah P. AU - Li, Ping AU - Di, Chunhui AU - Brown, Talmage AU - Salva, Maja Z. AU - Li, Songtao AU - Bird, Andrew AU - Yan, Zhen AU - Auten, Richard AU - Hauschka, Stephen D. AU - Koeberl, Dwight D. T2 - MOLECULAR THERAPY AB - Glycogen storage disease type II (Pompe disease; MIM 232300) stems from the deficiency of acid alpha-glucosidase (GAA; acid maltase; EC 3.2.1.20), which primarily involves cardiac and skeletal muscles. An adeno-associated virus 2/8 (AAV2/8) vector containing the muscle creatine kinase (MCK) (CK1) reduced glycogen content by approximately 50% in the heart and quadriceps in GAA-knockout (GAA-KO) mice; furthermore, an AAV2/8 vector containing the hybrid alpha-myosin heavy chain enhancer-/MCK enhancer-promoter (MHCK7) cassette reduced glycogen content by >95% in heart and >75% in the diaphragm and quadriceps. Transduction with an AAV2/8 vector was higher in the quadriceps than in the gastrocnemius. An AAV2/9 vector containing the MHCK7 cassette corrected GAA deficiency in the distal hindlimb, and glycogen accumulations were substantially cleared by human GAA (hGAA) expression therein; however, the analogous AAV2/7 vector achieved much lower efficacy. Administration of the MHCK7-containing vectors significantly increased striated muscle function as assessed by increased Rotarod times at 18 weeks after injection, whereas the CK1-containing vector did not increase Rotarod performance. Importantly, type IIb myofibers in the extensor digitalis longus (EDL) were transduced, thereby correcting a myofiber type that is unresponsive to enzyme replacement therapy. In summary, AAV8 and AAV9-pseudotyped vectors containing the MHCK7 regulatory cassette achieved enhanced efficacy in Pompe disease mice. DA - 2008/8// PY - 2008/8// DO - 10.1038/mt.2008.133 VL - 16 IS - 8 SP - 1366-1371 SN - 1525-0016 ER - TY - JOUR TI - Compendium of veterinary standard precautions for zoonotic disease prevention in veterinary personnel - National Association of State Public Health Veterinarians Veterinary Infection Control Committee 2008 AU - Elchos, B. L. AU - Scheftel, J. M. AU - Cherry, B. AU - DeBess, E. E. AU - Hopkins, S. G. AU - Levine, J. F. AU - Williams, C. J. AU - Bell, M. R. AU - Dvorak, G. D. AU - Flora, C. A. AU - Hofmann, J. AU - Pavlin, B. I. AU - Samples, O. M. AU - Snow, J. L. AU - Stinson-Dixon, R. E. T2 - Journal of the American Veterinary Medical Association DA - 2008/// PY - 2008/// VL - 233 IS - 3 SP - 415-432 ER - TY - JOUR TI - Analysis of sodium carboxymethylcellulose administration and related factors associated with postoperative colic and survival in horses with small intestinal disease AU - Fogle, Callie A. AU - Gerard, Mathewp. AU - Elce, Yvonne A. AU - Little, Dianne AU - Morton, Alison J. AU - Correa, Maria T. AU - Blikslager, Anthony T. T2 - VETERINARY SURGERY AB - Objective— To analyze the effect of the intraoperative use of sodium carboxymethylcellulose (CBMC) and related perioperative factors on postoperative colic and survival in horses that had abdominal surgery for colic. Study Design— Retrospective study. Animals— Horses (n=203) that had surgery for small intestinal disease; 33 horses had intraoperative administration of CBMC. Methods— Information was obtained from medical records for 170 horses that had surgery for colic before use of CBMC and 33 horses that had intraoperative CBMC. Kaplan–Meier survival curves were used to estimate median survival time and a Cox proportional hazards model was used to estimate the hazard ratio for the effect of CBMC and other perioperative variables on survival. Results— Seventy‐five percent of horses administered CBMC survived to 180 days, whereas 75% of untreated horses survived 8 days (median survival time=18 days). Horses not administered CBMC were twice as likely to die compared with horses administered CBMC. Horses that had postoperative ileus (POI) were 1.4 times more likely to die than horses without ileus. Similarly, horses with signs of colic after surgery were 1.3 times more likely to die than horses without postoperative signs of colic. Conclusions— CBMC administration is seemingly protective against death and prolongs survival when used intraoperatively in horses with small intestine disease, particularly horses with postoperative colic or POI. Both POI and colic increased risk of death after surgery. Clinical Relevance— Intraoperative administration of CBMC in horses that have surgery for small intestinal disease may improve survival, possibly by reducing early adhesion formation. DA - 2008/8// PY - 2008/8// DO - 10.1111/j.1532-950X.2008.00420.x VL - 37 IS - 6 SP - 558-563 SN - 1532-950X ER - TY - JOUR TI - Quantification of chemical mixture interactions modulating dermal absorption using a multiple membrane fiber array AU - Baynes, Ronald E. AU - Xia, Xin Rui AU - Irman, Mudassar AU - Riviere, Jim E. T2 - CHEMICAL RESEARCH IN TOXICOLOGY AB - Dermal exposures to chemical mixtures can potentially increase or decrease systemic bioavailability of toxicants in the mixture. Changes in dermal permeability can be attributed to changes in physicochemical interactions between the mixture, the skin, and the solute of interest. These physicochemical interactions can be described as changes in system coefficients associated with molecular descriptors described by Abraham's linear solvation energy relationship (LSER). This study evaluated the effects of chemical mixtures containing either a solvent (ethanol) or a surfactant (sodium lauryl sulfate, SLS) on solute permeability and partitioning by quantifying changes in system coefficients in skin and a three-membrane-coated fiber (MCF) system, respectively. Regression analysis demonstrated that changes in system coefficients in skin were strongly correlated ( R2 = 0.89-0.98) to changes in system coefficients in the three-membrane MCF array with mixtures containing either 1% SLS or 50% ethanol. The PDMS fiber appeared to play a significant role (R2 = 0.84-0.85) in the MCF array in predicting changes in solute permeability, while the WAX fiber appeared to contribute less (R2 = 0.59-0.77) to the array than the other two fibers. On the basis of changes in system coefficients that are part of a LSER, these experiments were able to link physicochemical interactions in the MCF with those interactions in skin when either system is exposed to 1% SLS or 50% ethanol. These experiments further demonstrated the utility of a MCF array to adequately predict changes in dermal permeability when skin is exposed to mixtures containing either a surfactant or a solvent and provide some insight into the nature of the physiochemical interactions that modulate dermal absorptions. DA - 2008/3// PY - 2008/3// DO - 10.1021/tx7002118 VL - 21 IS - 3 SP - 591-599 SN - 1520-5010 ER - TY - JOUR TI - Pharmacokinetics of melamine in pigs following intravenous administration AU - Baynes, R. E. AU - Smith, Geof AU - Mason, S. E. AU - Barrett, E. AU - Barlow, B. M. AU - Riviere, J. E. T2 - Food and Chemical Toxicology AB - Melamine-contaminated pet food was recently added as a supplement to livestock feed. There is little or no information concerning the pharmacokinetics of melamine in livestock, and the aim of this study was to obtain pharmacokinetic parameters for this contaminant in pigs. Melamine was administered intravenously to five weanling pigs at a dose of 6.13 mg/kg and plasma samples were collected over 24 h, extracted for melamine, and then analyzed by HPLC-UV. The data was shown to best fit a one-compartment model with melamine's half-life of 4.04 (+/- 0.37) h, clearance of 0.11 (+/- 0.01) L/h/kg, and volume of distribution of 0.61 (+/- 0.04) L/kg. These data are comparable to the only mammalian study in rats and suggests that melamine is readily cleared by the kidney and there is unlikely to be significant tissue binding. Further tissue residue studies are required to assess the depletion kinetics of this contaminant in the pig which will determine whether residue levels in the kidney should be of public health concern if pigs were exposed to a similar dose. DA - 2008/// PY - 2008/// DO - 10.1016/j.fct.2007.11.013 VL - 46 IS - 3 SP - 1196–1200 ER - TY - JOUR TI - Fatty acid composition of wild anthropoid primate milks AU - Milligan, Lauren A. AU - Rapoport, Stanley I. AU - Cranfield, Michael R. AU - Dittus, Wolfgang AU - Glander, Kenneth E. AU - Oftedal, Olav T. AU - Power, Michael L. AU - Whittier, Christopher A. AU - Bazinet, Richard P. T2 - COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY AB - Fatty acids in milk reflect the interplay between species-specific physiological mechanisms and maternal diet. Anthropoid primates (apes, Old and New World monkeys) vary in patterns of growth and development and dietary strategies. Milk fatty acid profiles also are predicted to vary widely. This study investigates milk fatty acid composition of five wild anthropoids (Alouatta palliata, Callithrix jacchus, Gorilla beringei beringei, Leontopithecus rosalia, Macaca sinica) to test the null hypothesis of a generalized anthropoid milk fatty acid composition. Milk from New and Old World monkeys had significantly more 8:0 and 10:0 than milk from apes. The leaf eating species G. b. beringei and A. paliatta had a significantly higher proportion of milk 18:3n-3, a fatty acid found primarily in plant lipids. Mean percent composition of 22:6n-3 was significantly different among monkeys and apes, but was similar to the lowest reported values for human milk. Mountain gorillas were unique among anthropoids in the high proportion of milk 20:4n-6. This seems to be unrelated to requirements of a larger brain and may instead reflect species-specific metabolic processes or an unknown source of this fatty acid in the mountain gorilla diet. DA - 2008/1// PY - 2008/1// DO - 10.1016/j.cbpb.2007.08.006 VL - 149 IS - 1 SP - 74-82 SN - 1879-1107 KW - anthropoid KW - fatty acid KW - milk composition KW - primate ER - TY - JOUR TI - FARAD digest - Extralabel use of nonsteroidal anti-inflammatory drugs in cattle AU - Smith, G. W. AU - Davis, J. L. AU - Tell, L. A. AU - Webb, A. I. AU - Riviere, J. E. T2 - Journal of the American Veterinary Medical Association DA - 2008/// PY - 2008/// VL - 232 IS - 5 SP - 697-701 ER - TY - JOUR TI - Epicutaneous sensitization with Dermatophagoides farinae induces generalized allergic dermatitis and elevated mite-specific immunoglobulin E levels in a canine model of atopic dermatitis AU - Pucheu-Haston, C. M. AU - Jackson, H. A. AU - Olivry, T. AU - Dunston, S. M. AU - Hammerberg, B. T2 - CLINICAL AND EXPERIMENTAL ALLERGY AB - Summary Background Atopic dermatitis (AD) is a cutaneous hypersensitivity associated with elevated levels of antigen‐specific IgE, commonly to house dust mites (HDMs). It remains controversial as to whether sensitization and clinical disease are induced by cutaneous exposure to HDM. Objectives The objectives of this study were to determine whether repeated applications of Dermatophagoides farinae slurry to intact skin of Maltese–Beagle atopic (MAB) dogs would result in the development of clinical signs or lesions resembling spontaneous canine AD, to determine whether repeated slurry applications would induce elevations in mite‐specific IgE and/or IgG, and to determine whether mite antigens could be demonstrated within the dermis of application sites. Methods Dogs received weekly slurry applications to the axilla and groin, and were patch tested at 120 days, or were patch tested at days 1, 60 and 120, but did not receive further slurry applications. Skin biopsies and serum samples were obtained on days 1, 60 and 120. Results Pruritic dermatitis was seen in all dogs by day 60. D. farinae ‐specific IgE was elevated by day 60. Histologic examination of early application sites revealed mild, mononuclear perivascular dermatitis. Later application sites were characterized by a dense inflammatory infiltrate and oedema in both the dermis and the epidermis. Immunofluorescent staining confirmed the presence of D. farinae antigens in the dermis. Conclusions This study demonstrated that epicutaneous application of HDM slurry to MAB dogs results in elevations of HDM‐specific IgE, localized and generalized pruritic dermatitis resembling spontaneous canine AD, and histologic changes typical of IgE‐driven inflammation. We feel that these results suggest that epicutaneous exposure to allergen may play an important role during both the sensitization and the perpetuation of AD, and provide support for the use of a canine model in the investigation of the pathogenesis of AD. DA - 2008/4// PY - 2008/4// DO - 10.1111/j.1365-2222.2008.02949.x VL - 38 IS - 4 SP - 667-679 SN - 0954-7894 KW - allergy KW - animal models KW - atopic dermatitis KW - atopy KW - Dermatophagoides farinae KW - dogs KW - skin (dermatology) immunology ER - TY - JOUR TI - Assessment of the Effect of Varying Soil Organic Matter Content on the Bioavailability of Malathion to the Common Nightcrawler, Lumbricus terrestris L. AU - Henson-Ramsey, Heather AU - Shea, Damian AU - Levine, Jay F. AU - Kennedy-Stoskopf, Suzanne AU - Taylor, Sharon K. AU - Stoskopf, Michael K. T2 - Bulletin of Environmental Contamination and Toxicology DA - 2008/1/19/ PY - 2008/1/19/ DO - 10.1007/s00128-007-9349-6 VL - 80 IS - 3 SP - 220-224 J2 - Bull Environ Contam Toxicol LA - en OP - SN - 0007-4861 1432-0800 UR - http://dx.doi.org/10.1007/s00128-007-9349-6 DB - Crossref KW - earthworm KW - malathion KW - bioavailability KW - soil organic matter ER - TY - JOUR TI - Sulfamethazine water medication pharmacokinetics and contamination in a commercial pig production unit AU - Mason, Sharon E. AU - Baynes, Ronald E. AU - Buur, Jennifer L. AU - Riviere, Jim E. AU - Almond, Glen W. T2 - JOURNAL OF FOOD PROTECTION AB - Sulfamethazine is often used to treat disease in the swine industry. Sulfamethazine is available as water or feed medication and historically (over the past 40 years) has been associated with residue violations in both the United States and Europe. Despite sulfamethazine's approval for use as a water medication, little research on the pharmacokinetics of the water formulation is available. Therefore, a pilot study was performed to determine the plasma levels of an approved sulfamethazine water medication. Plasma levels in pigs treated with an oral bolus (250 mg/kg), which is equivalent to the total drug consumed within a 24-h period, achieved therapeutic concentrations (50 microg/ml). Noncompartmental-based pharmacokinetic model parameters for clearance, half-life, and volume of distribution were consistent with previously published values in swine. However, the above treatment resulted in exposure of pen mates to sulfamethazine at levels currently above tolerance (0.1 ppm). Using a physiologically based pharmacokinetic model, the treatment dose simulation was compared with observed plasma levels of treated pigs. Flexibility of the physiologically based pharmacokinetic model also allowed simulation of control-pig plasma levels to estimate contamination exposure. A simulated exposure to 0.15 mg/kg twice within approximately 8 h resulted in detectable levels of sulfamethazine in the control pigs. After initial exposure, a much lower dose of 0.059 mg/kg maintained the contamination levels above tolerance for at least 3 days. These results are of concern for producers and veterinarians, because in commercial farms, the entire barn is often treated,and environmental contamination could result in residues of an unknown duration. DA - 2008/3// PY - 2008/3// DO - 10.4315/0362-028X-71.3.584 VL - 71 IS - 3 SP - 584-589 SN - 1944-9097 ER - TY - JOUR TI - Detection of spring viraemia of carp virus (SVCV) by loop-mediated isothermal amplification (LAMP) in koi carp, Cyprinus carpio L AU - Shivappa, R B AU - Savan, R AU - Kono, T AU - Sakai, M AU - Emmenegger, E AU - Kurath, G AU - Levine, J F T2 - Journal of Fish Diseases AB - Abstract Spring viraemia of carp virus (SVCV) is a rhabdovirus associated with systemic illness and mortality in cyprinids. Several diagnostic tests are available for detection of SVCV. However, most of these tests are time consuming and are not well adapted for field‐based diagnostics. In this study, a diagnostic tool for SVCV detection based on reverse transcription loop‐mediated isothermal amplification (RT‐LAMP) has been developed. Based on the nucleotide sequence of the glycoprotein (G) gene of SVCV North Carolina (NC) isolate, four sets (each set containing two outer and two inner) of primers were designed. Temperature and time conditions were optimized to 65 °C and 60 min, respectively, for LAMP and RT‐LAMP using one primer set. In vitro specificity was evaluated using four different strains of fish rhabdoviruses and RT‐LAMP was found to be specific to SVCV. Serial dilutions of SVCV NC isolate was used to evaluate the in vitro sensitivity of RT‐LAMP. Sensitivity of the assays was similar to RT‐PCR and detected SVCV even at the lowest dilution of 10 1 TCID 50 mL −1 . The ability of RT‐LAMP to detect SVCV from infected carp was also tested and the assay detected SVCV from all infected fish. The isothermal temperature requirements, high specificity and sensitivity, and short incubation time of the RT‐LAMP assay make it an excellent choice as a field diagnostic test for SVCV. DA - 2008/4// PY - 2008/4// DO - 10.1111/j.1365-2761.2007.00894.x VL - 31 IS - 4 SP - 249-258 J2 - J Fish Diseases LA - en OP - SN - 0140-7775 1365-2761 UR - http://dx.doi.org/10.1111/j.1365-2761.2007.00894.x DB - Crossref KW - detection KW - koi carp KW - loop mediated isothermal amplification KW - spring viraemia of carp virus ER - TY - JOUR TI - Cytauxzoon felis infections are present in bobcats (Lynx rufus) in a region where cytauxzoonosis is not recognized in domestic cats AU - Birkenheuer, Adam J. AU - Marr, Henry S. AU - Warren, Camille AU - Acton, Anne E. AU - Mucker, Eric M. AU - Humphreys, Jan G. AU - Tucker, Melissa D. T2 - VETERINARY PARASITOLOGY AB - This study was performed to determine the prevalence of Cytauxzoon felis (C. felis) infections in bobcats (Lynx rufus) from a region where C. felis is recognized in domestic cats, North Carolina (NC), and a region where C. felis is not recognized in domestic cats, Pennsylvania (PA). Samples from NC (n=32) were obtained post-mortem via cardiac puncture from legally trapped bobcats. Samples from PA (n=70) were collected post-mortem onto Nobuto blood collecting strips by the PA Game Commission. Each sample was tested using a C. felis specific PCR assay as well as a PCR assay targeting host DNA to rule out the presence of PCR inhibitors. Three samples were excluded due to the presence of PCR inhibitors. Thirty-three percent (10/30) of the samples from NC and 7% (5/69) of the samples from PA tested positive for the presence of C. felis. The proportion of C. felis positive bobcats from NC was significantly different than that from PA (P<0.005). Despite the lower prevalence of C. felis infections in bobcats from PA this finding is unique and indicates the potential for C. felis infections in domestic cats in the northeastern USA if the appropriate tick vectors are present. Veterinary practitioners in PA should be on alert for cytauxzoonosis in domestic cats. Further studies about the epidemiology and transmission of C. felis infections among both domestic cats and bobcats are needed. DA - 2008/5/6/ PY - 2008/5/6/ DO - 10.1016/j.vetpar.2008.01.020 VL - 153 IS - 1-2 SP - 126-130 SN - 1873-2550 KW - cytauxzoonosis KW - piroplasmosis KW - Cytauxzoon felis KW - bobcats KW - PCR KW - tick ER - TY - JOUR TI - Comparison of two indirect techniques for local delivery of a high dose of an antimicrobial in the distal portion of forelimbs of horses AU - Errico, Jason A. AU - Trumble, Troy N. AU - Bueno, Aloisio C. D. AU - Davis, Jennifer L. AU - Brown, Murray P. T2 - AMERICAN JOURNAL OF VETERINARY RESEARCH AB - Abstract Objective —To compare isolated limb retrograde venous injection (ILRVI) and isolated limb infusion (ILI) for delivery of amikacin to the synovial fluid of the distal interphalangeal and metacarpophalangeal joints and to evaluate the efficacy of use of an Esmarch tourniquet in standing horses. Animals —6 healthy adult horses. Procedures —Horses were randomly assigned in a crossover design. In ILRVI, the injection consisted of 1 g of amikacin diluted to a total volume of 60 mL administered during a 3-minute period. In ILI, the infusion consisted of 1 g of amikacin diluted to 40 mL administered during a 3-minute period followed by administration of boluses of diluent (82 mL total) to maintain vascular pressure. During ILI, the infusate and blood were circulated from the venous to the arterial circulation in 5-mL aliquots. Synovial fluid and serum samples were obtained to determine maximum amikacin concentrations and tourniquet leakage, respectively. Results —Both techniques yielded synovial concentrations of amikacin > 10 times the minimum inhibitory concentration (MIC) for 90% of isolates (80 μg/mL) and > 10 times the MIC breakpoint (160 μg/mL) of amikacin-susceptible bacteria reported to cause septic arthritis in horses. These values were attained for both joints for both techniques. Esmarch tourniquets prevented detectable loss of amikacin to the systemic circulation for both techniques. Conclusions and Clinical Relevance —Both techniques reliably achieved synovial fluid concentrations of amikacin consistent with concentration-dependent killing for bacteria commonly encountered in horses with septic arthritis. Esmarch tourniquets were effective for both delivery techniques in standing horses. DA - 2008/3// PY - 2008/3// DO - 10.2460/ajvr.69.3.334 VL - 69 IS - 3 SP - 334-342 SN - 0002-9645 ER - TY - JOUR TI - Application of linear solvation energy relationships to a custom-made polyaniline solid-phase microextraction fiber and three commercial fibers AU - Yeatts, James L., Jr. AU - Baynes, Ronald E. AU - Xia, Xin-Rui AU - Riviere, Jim E. T2 - JOURNAL OF CHROMATOGRAPHY A AB - The term linear solvation energy relationships, LSERs, is considered to be a specific subset of a larger group of thermodynamic relationships called linear free energy relationships. Overall, the LSERs model represents a three-step thermodynamic process. The most recently accepted notation for the LSER equation, proposed by Abraham is given as follows:SP=c+eE+sS+aA+bB+vVwhere SP is any free energy related property of a solute, such as log K, and each term in the equation represents a specific type of chemical interaction. In this work, LSERs were applied to a custom-made polyaniline (PANI) solid-phase microextraction fiber and three commercial fibers immersed in water in order to aid in the assessment of a diverse series of solutes’ partitioning behavior. By experimentally determining the log K for a series of solutes with known solute descriptors (E, S, A, B, and V) and performing multi-linear regression, the unknown system coefficients (e, s, a, b, and v) were obtained. The sign and magnitude of the system coefficients reflect the relative strengths of chemical interactions that affect partitioning between the two phases (fiber and water). The LSER study showed that the system properties having the greatest influence on log K were ease of cavity formation and hydrogen bond donating ability. The differences in dipolarity/polarizability as well as in hydrogen bond accepting ability further showed that all four fibers offer a unique environment for solute partitioning. The PANI fiber may offer greater flexibility in the choice of fibers to use for solid-phase microextraction. DA - 2008/4/25/ PY - 2008/4/25/ DO - 10.1016/j.chroma.2008.02.057 VL - 1188 IS - 2 SP - 108-117 SN - 1873-3778 KW - polyaniline KW - solid-phase microextraction KW - biocides KW - linear solvation energy relationships KW - metal core fiber KW - partition coefficients ER - TY - JOUR TI - AAV vector-mediated reversal of hypoglycemia in canine and murine glycogen storage disease type Ia AU - Koeberl, Dwight D. AU - Pinto, Carlos AU - Sun, Baodong AU - Li, Songtao AU - Kozink, Daniel M. AU - Benjamin, Daniel K., Jr. AU - Demaster, Amanda K. AU - Kruse, Meghan A. AU - Vaughn, Valerie AU - Hillman, Steven AU - Bird, Andrew AU - Jackson, Mark AU - Brown, Talmage AU - Kishnani, Priya S. AU - Chen, Yuan-Tsong T2 - MOLECULAR THERAPY AB - Glycogen storage disease type Ia (GSD-Ia) profoundly impairs glucose release by the liver due to glucose-6-phosphatase (G6Pase) deficiency. An adeno-associated virus (AAV) containing a small human G6Pase transgene was pseudotyped with AAV8 (AAV2/8) to optimize liver tropism. Survival was prolonged in 2-week-old G6Pase (-/-) mice by 600-fold fewer AAV2/8 vector particles (vp), in comparison to previous experiments involving this model (2 x 10(9) vp; 3 x 10(11) vp/kg). When the vector was pseudotyped with AAV1, survival was prolonged only at a higher dose (3 x 10(13) vp/kg). The AAV2/8 vector uniquely prevented hypoglycemia during fasting and fully corrected liver G6Pase deficiency in GSD-Ia mice and dogs. The AAV2/8 vector has prolonged survival in three GSD-Ia dogs to >11 months, which validated this strategy in the large animal model for GSD-Ia. Urinary biomarkers, including lactate and 3-hydroxybutyrate, were corrected by G6Pase expression solely in the liver. Glycogen accumulation in the liver was reduced almost to the normal level in vector-treated GSD-Ia mice and dogs, as was the hepatocyte growth factor (HGF) in GSD-Ia mice. These preclinical data demonstrated the efficacy of correcting hepatic G6Pase deficiency, and support the further preclinical development of AAV vector-mediated gene therapy for GSD-Ia. DA - 2008/4// PY - 2008/4// DO - 10.1038/mt.2008.15 VL - 16 IS - 4 SP - 665-672 SN - 1525-0016 ER - TY - JOUR TI - What is your diagnosis? Particulate material in peritoneal fluid from a dog AU - Renschler, Janelle AU - Tarigo, Jaime AU - Neel, Jennifer AU - Grindem, Carol T2 - VETERINARY CLINICAL PATHOLOGY AB - Abstract: 9‐year‐old castrated male Greyhound dog was presented for evaluation of vomiting and lethargy of 1‐week duration. On physical examination, the dog was febrile and dehydrated with a tense abdomen and petechial hemorrhages. Clinicopathologic abnormalities included relative polycythemia, mild lymphopenia with reactive lymphocytes, hypoalbuminemia, hypocholesterolemia, hyperbilirubinemia, increased ALP, mild hypokalemia, hyperamylasemia, hyperlipasemia, increased D‐dimer concentration, and hyperfibrinogenemia. Cytologic evaluation of peritoneal fluid revealed marked suppurative inflammation with intracellular barium sulfate particles. The day before presentation, the referring veterinarian had administered oral barium sulfate in an upper gastrointestinal contrast study. Radiographs revealed free contrast material in the peritoneal cavity, consistent with gastrointestinal perforation, and leakage of contrast material. Abdominal exploratory surgery revealed a mid‐jejunal perforation and a hepatic nodule. Histopathologic diagnosis of the jejunal and liver lesions was T‐cell lymphoma. The patient recovered well postoperatively and received chemotherapy for treatment of lymphoma. Most commercial barium sulfate preparations contain relatively uniform, weakly birefringent, pale yellow particles <1 μm in diameter. Because barium sulfate is found occasionally in clinical specimens, cytopathologists should be familiar with its cytologic appearance. DA - 2008/3// PY - 2008/3// DO - 10.1111/j.1939-165X.2008.00006.x VL - 37 IS - 1 SP - 129-131 SN - 1939-165X UR - http://www.scopus.com/inward/record.url?eid=2-s2.0-44449114150&partnerID=MN8TOARS KW - abdominal fluid KW - barium sulfate KW - dog KW - lymphoma KW - peritonitis KW - radiographic contrast ER - TY - JOUR TI - The effect of quicklime (CaO) on litter condition and broiler performance AU - Ruiz, V. AU - Ruiz, D. AU - Gernat, A. G. AU - Grimes, J. L. AU - Murillo, J. G. AU - Wineland, M. J. AU - Anderson, K. E. AU - Maguire, R. O. T2 - POULTRY SCIENCE AB - High levels of phosphorus and pathogens in runoff are 2 major concerns following manure applications to fields. Phosphorus losses from fields following manure applications have been linked to the solubility of phosphorus in manure; therefore, by decreasing manure phosphorus solubility, a decrease in phosphorus loss in runoff should be apparent. The objective of this research was to develop a process using quicklime that would result in reduced phosphorus solubility and bacteria counts in broiler litter. The 4 litter treatments evaluated were T1, new wood shavings without the addition of quicklime; T2, used, untreated broiler litter; T3, used litter with 10% quicklime (based on the weight of the litter); and T4, used litter with 15% quicklime (based on the weight of the litter). Body weight, cumulative feed consumption, and feed conversion (feed: BW) were determined on a weekly basis through 42 d of age. Mortality was recorded daily. Carcass weights and percentages of carcass yield without giblets were determined prechill. Litter pH, total phosphorus, nitrogen, soluble phosphorus, litter moisture (%), and total plate counts were measured for each litter treatment on d 7 and 42 after bird placement. No significant differences were found for BW, feed consumption, feed conversion, mortality, carcass weight, or carcass yield. No breast or footpad blisters were observed. On d 7, 15% quicklime had higher (P < 0.001) pH (11.2) when compared with the other treatments. Percentages of phosphorus and nitrogen were lower (P < 0.001) for new wood shavings in comparison with the used litter treatments. Soluble phosphorus (ppm) was lower (P < 0.001) for 15% quicklime (2.75) when compared with new wood shavings (42.2), untreated broiler litter (439.2), and 10% quicklime (35.0). Although not significant, 15% quicklime had lower total plate counts (cfu/g) in comparison with the other treatments on d 1 and 10 postmixing and at 7 d after bird placement. Litter conditions on d 42 after bird placement were similar. We concluded that the use of quicklime as a treatment for broiler litter would initially reduce nitrogen and soluble phosphorus and bacteria counts without negatively affecting bird productivity. DA - 2008/5// PY - 2008/5// DO - 10.3382/ps.2007-00101 VL - 87 IS - 5 SP - 823-827 SN - 0032-5791 KW - broiler KW - litter treatment KW - quicklime KW - phosphorus KW - litter pathogen ER - TY - JOUR TI - Vehicle effects on in vitro transdermal absorption of sevoflurane in the bullfrog, Rana catesbeiana AU - Ardente, Amanda J. AU - Barlow, Beth M. AU - Burns, Patrick AU - Goldman, Rebecca AU - Baynes, Ronald E. T2 - ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY AB - The experimental objectives were to identify a vehicle which produces a homogenous formulation when combined with the anesthetic solution sevoflurane and understand the dermal absorption of sevoflurane in silastic membranes and amphibian skin in vitro utilizing a flow-through diffusion system. Seven vehicles were evaluated in varying ratios with 5 formulations resulting in the desired homogenous consistency for practical application. Sevoflurane diffusion across silastic membranes was influenced by pluronic/lecithin organogel (PLO), pluronic F 127 20% gel, and sterile lube. Flux and permeability across silastic membranes were significantly greater in sterile lube than in the other formulations. While no significant vehicle effects were observed in bullfrog skin, the flux-time profiles suggest that sevoflurane diffusion in bullfrog skin may be positively influenced by PLO. Future in vivo studies are required to assess sevoflurane retention after removal of these formulations to more accurately control the plane of anesthesia in amphibians. DA - 2008/5// PY - 2008/5// DO - 10.1016/j.etap.2007.12.001 VL - 25 IS - 3 SP - 373-379 SN - 1382-6689 KW - amphibian KW - dermal KW - sevofluame KW - in vitro KW - permeability ER - TY - JOUR TI - Vector transmission of Bartonella species with emphasis on the potential for tick transmission AU - Billeter, S. A. AU - Levy, M. G. AU - Chomel, B. B. AU - Breitschwerdt, E. B. T2 - MEDICAL AND VETERINARY ENTOMOLOGY AB - Bartonella species are gram-negative bacteria that infect erythrocytes, endothelial cells and macrophages, often leading to persistent blood-borne infections. Because of the ability of various Bartonella species to reside within erythrocytes of a diverse number of animal hosts, there is substantial opportunity for the potential uptake of these blood-borne bacteria by a variety of arthropod vectors that feed on animals and people. Five Bartonella species are transmitted by lice, fleas or sandflies. However, Bartonella DNA has been detected or Bartonella spp. have been cultured from numerous other arthropods. This review discusses Bartonella transmission by sandflies, lice and fleas, the potential for transmission by other vectors, and data supporting transmission by ticks. Polymerase chain reaction (PCR) or culture methods have been used to detect Bartonella in ticks, either questing or host-attached, throughout the world. Case studies and serological or molecular surveys involving humans, cats and canines provide indirect evidence supporting transmission of Bartonella species by ticks. Of potential clinical relevance, many studies have proposed co-transmission of Bartonella with other known tick-borne pathogens. Currently, critically important experimental transmission studies have not been performed for Bartonella transmission by many potential arthropod vectors, including ticks. DA - 2008/3// PY - 2008/3// DO - 10.1111/j.1365-2915.2008.00713.x VL - 22 IS - 1 SP - 1-15 SN - 1365-2915 KW - Bartonella species KW - arthropods KW - DNA KW - PCR KW - ticks KW - vector competence KW - vector potential ER - TY - JOUR TI - Lack of detection of circulating skin-specific IgE autoantibodies in dogs with moderate or severe atopic dermatitis AU - Olivry, Thierry AU - Dunston, Stanley A. AU - Pluchino, Kristen AU - Porter, Kyleigh AU - Hammerberg, Bruce T2 - VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY AB - Human patients with atopic dermatitis (AD) commonly exhibit IgE reactivity to cutaneous self-antigens. The presence of serum IgE autoantibodies appears to correlate with disease severity, and it is suspected to reflect or contribute to tissue damage. The objective of this study was to determine whether IgE autoantibodies specific for cutaneous antigens could be detected in the serum of dogs with AD. Serum was collected from 19 dogs with untreated moderate to severe AD and four specific-pathogen free (SPF) dogs. Indirect immunofluorescence was performed using normal canine skin collected at four different locations (concave ear, nose, medial thigh and lateral thorax), while Western immunoblotting was done using normal canine ear pinna epidermal and dermal extracts and reducing conditions. In both methods, IgE was detected using a monoclonal antibody specific for heat stable epitopes of canine IgE. At 1:10 dilution, specific IgE autoantibodies against cutaneous autoantigens were not detected, with either method, in AD and SPF canine sera. Either IgE autoreactivity is not associated with moderate to severe AD in dogs, or the methods employed herein were not sensitive enough to permit IgE autoantibody detection. DA - 2008/3/15/ PY - 2008/3/15/ DO - 10.1016/j.vetimm.2007.11.003 VL - 122 IS - 1-2 SP - 182-187 SN - 1873-2534 KW - allergy KW - atopy KW - autoimmunity KW - canine KW - IgE KW - skin ER - TY - JOUR TI - Cyclooxygenase expression and prostanoid production in pyloric and duodenal mucosae in dogs after administration of nonsteroidal anti-inflammatory drugs AU - Wooten, Jenna G. AU - Blikslager, Anthony T. AU - Ryan, Kathleen A. AU - Marks, Steve L. AU - Mac Law, J. AU - Lascelles, Duncan X. T2 - AMERICAN JOURNAL OF VETERINARY RESEARCH AB - Abstract Objective —To assess cyclooxygenase (COX) expression and prostanoid concentrations in pyloric and duodenal mucosae of dogs after administration of nonsteroidal anti-inflammatory drugs (NSAIDs). Animals —8 healthy dogs. Procedures —Each dog received carprofen (4.4 mg/kg, q 24 h), deracoxib (2 mg/kg, q 24 h), aspirin (10 mg/kg, q 12 h), and placebo (1 dog treat, q 24 h) orally for 3 days (4-week interval between treatments). Before study commencement (baseline) and on day 3 of each treatment, pyloric and duodenal mucosal appearance was assessed endoscopically and biopsy specimens were obtained for histologic examination. Cyclooxygenase-1 and COX-2 protein expressions were assessed via western blotting, and prostanoid concentrations were measured via ELISAs. An ANOVA was used to analyze data. Results —Treatments had no effect on mucosal appearance and ulceration was not evident histologically. In pyloric and duodenal mucosae, COX-1 expression was unaffected by treatments. Cyclooxygenase-2 expression remained unchanged in pyloric mucosa; in duodenal mucosa, aspirin significantly increased COX-2 expression, compared with effects of deracoxib and carprofen. At baseline, total prostaglandin and thromboxane B 2 concentrations in pyloric mucosa were significantly greater than those in duodenal mucosa. Aspirin significantly decreased both prostanoid concentrations in both mucosal tissues, compared with other treatments. In pyloric mucosa, carprofen administration significantly decreased total prostaglandin and thromboxane B 2 concentrations, compared with deracoxib administration. Conclusions and Clinical Relevance —In dogs, prostanoid synthesis was greater in pyloric mucosa than it was in duodenal mucosa. Nonselective NSAIDs significantly decreased prostanoid concentrations in these mucosae, compared with the effects of a selective COX-2 NSAID. DA - 2008/4// PY - 2008/4// DO - 10.2460/ajvr.69.4.457 VL - 69 IS - 4 SP - 457-464 SN - 1943-5681 ER - TY - JOUR TI - Cloning of feline FOXP3 and detection of expression in CD4+CD25+ regulatory T cells AU - Lankford, Susan AU - Petty, Christopher AU - La Voy, Alora AU - Reckling, Stacie AU - Tompkins, Wayne AU - Dean, Gregg A. T2 - VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY AB - Regulatory T cells (Treg) are increased and directly infected by feline immunodeficiency virus (FIV) and likely play a role in other feline autoimmune, neoplastic, and infectious diseases. Phenotypically, Treg are best characterized by surface expression of CD4 and CD25 and intranuclear expression of the forkhead transcription factor Foxp3. Our objective was to clone and sequence feline FOXP3 for the purpose of developing assays to enhance studies of feline Treg. We determined the feline FOXP3 is 1293 nucleotides in length and codes for a protein that shares high homology to other species. A splice variant devoid of exon 2 was also identified. A real-time PCR assay was developed and used to show Foxp3 mRNA expression occurs primarily in CD4+CD25+ T cells. Two cross-reacting antibodies were identified by immunocytochemical staining of HEK293 cells transfected with feline FOXP3. The antibody labeling confirmed the nuclear localization of the protein. A flow cytometric assay was also validated and used to correlate the phenotypic and functional characteristics of feline Treg induced by treatment of lymph node lymphocytes with flagellin or LPS in combination with mitogen or IL2. Together, these studies provide useful tools to further investigate Foxp3 and Tregs in cats. DA - 2008/3/15/ PY - 2008/3/15/ DO - 10.1016/j.vetimm.2007.11.007 VL - 122 IS - 1-2 SP - 159-166 SN - 1873-2534 KW - Foxp3 KW - feline KW - Treg KW - regulatory T cells KW - toll-like receptors ER - TY - JOUR TI - Expression of CDK4 or CDK2 in mouse oral cavity is retained in adult pituitary with distinct effects on tumorigenesis AU - Macias, Everardo AU - Marva, Paula L. Miliani AU - Senderowicz, Adrian AU - Cullen, John AU - Rodriguez-Puebla, Marcelo L. T2 - CANCER RESEARCH AB - The keratin 5 (K5) promoter drives transgenic expression to the basal cell layer of stratified epithelia. Surprisingly, analysis of K5CDK4 and K5CDK2 transgenic mouse embryos showed CDK4 and CDK2 expression not only in the expected tissues, but also in the adenohypophysis. This organ is derived from an upwards growth of the primitive oropharynx, a K5-expressing tissue. We show that transgenic expression of CDKs in the embryonic oral ectoderm is specifically retained in undifferentiated cells from the pars intermedia of the adenohypophysis. Interestingly, we found that K5CDK4 mice show a decreased number of pituitary stem cells, even though CDK4 is not expressed in the stem cells but in transit-amplifying (TA)-like cells. Interestingly, CDK4-expressing cells, but not CDK2-expressing cells, strongly synergize with lack of p27(Kip1) to generate pituitary carcinomas that appear with shortened latency and are drastically more aggressive than those arising in p27(-/-) mice. Thus, we show that deregulation of CDK expression in the primitive oral epithelium plays a unique function, providing a selective advantage that gives rise to transgene-positive TA-like pituitary cells. Furthermore, retention of CDK4 in these TA-like pituitary cells synergizes with loss of p27(Kip1) to induce pituitary adenocarcinomas. This model suggests that forced expression of CDK4 sensitizes cells and synergizes with a second change resulting in tumor development. DA - 2008/1/1/ PY - 2008/1/1/ DO - 10.1158/0008-5472.CAN-07-2461 VL - 68 IS - 1 SP - 162-171 SN - 0008-5472 ER - TY - JOUR TI - Transforming growth factor-beta/transforming growth factor-beta RII signaling may regulate CD4(+)CD25(+) T-regulatory cell homeostasis and suppressor function in feline AIDS lentivirus infection AU - Petty, Christopher S. AU - Tompkins, Mary B. AU - Tompkins, Wayne A. T2 - JAIDS-JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES AB - We have reported that CD4+CD25+ T-regulatory (Treg) cells are fully activated for suppressor function in feline immunodeficiency virus (FIV)-infected cats. Studies have suggested that surface transforming growth factor-beta (TGFbeta; membrane TGFbeta [mTGFbeta]) is a feature of activated CD4+CD25+ Treg cells and may play a role in Treg homeostasis and suppressor function. Herein, we explore the role of TGFbeta in feline Treg homeostasis and suppressor function and what effect FIV infection of cats might have on these processes. Stimulation of CD4+CD25- T helper (Th) cells with Concanavalin A (ConA) plus TGFbeta converts them to Treg-like cells capable of suppressor function. Reverse-transcription polymerase chain reaction and flow cytometry revealed that these ConA/TGFbeta-converted Treg cells upregulate Foxp3 and mTGFbeta. ConA stimulation of CD4+CD25- T cells upregulates TGFbeta receptor II (RII), and pretreatment of these cells with anti-TGFbeta-RII antibodies blocks the TGFbeta-induced conversion to Treg cells. Pretreatment of ConA/TGFbeta-converted Treg cells with anti-TGFbeta antibodies also abrogates their suppressor function, suggesting that Treg homeostasis and suppressor function may be mediated by mTGFbeta. Finally, we show that treatment of CD4+CD25+ mTGFbeta-positive Treg cells from FIV-infected cats with anti-TGFbeta antibodies or treatment of ConA-stimulated CD4+CD25- Th target cells with anti-TGFbeta-RII antibodies diminishes suppressor function. These data suggest that the recruitment of Treg cells from the Th pool and suppressor function of Treg cells are dependent on the TGFbeta/TGFbeta-RII signaling pathway and that this pathway is constitutively upregulated in asymptomatic chronically FIV-infected cats. DA - 2008/2/1/ PY - 2008/2/1/ DO - 10.1097/QAI.0b013e318160df70 VL - 47 IS - 2 SP - 148-160 SN - 1525-4135 KW - AIDS KW - immunodeficiency diseases KW - other animals KW - T lymphocytes ER - TY - JOUR TI - Pharmacokinetics of voriconazole after oral administration of single and multiple doses in African grey parrots (Psittacus erithacus timneh) AU - Flammer, Keven AU - Osborne, Julie A. Nettifee AU - Webb, Donna J. AU - Foster, Laura E. AU - Dillard, Stacy L. AU - Davis, Jennifer L. T2 - AMERICAN JOURNAL OF VETERINARY RESEARCH AB - Abstract Objective —To determine the pharmacokinetics and safety of orally administered voriconazole in African grey parrots. Animals —20 clinically normal Timneh African grey parrots ( Psittacus erithacus timneh ). Procedures —In single-dose trials, 12 parrots were each administered 6, 12, and 18 mg of voriconazole/kg orally and plasma concentrations of voriconazole were determined via high-pressure liquid chromatography. In a multiple-dose trial, voriconazole (18 mg/kg) was administered orally to 6 birds every 12 hours for 9 days; a control group (2 birds) received tap water. Treatment effects were assessed via observation, clinicopathologic analyses (3 assessments), and measurement of trough plasma voriconazole concentrations (2 assessments). Results —Voriconazole's elimination half-life was short (1.1 to 1.6 hours). Higher doses resulted in disproportional increases in the maximum plasma voriconazole concentration and area under the curve. Trough plasma voriconazole concentrations achieved in the multiple-dose trial were lower than those achieved after administration of single doses. Polyuria (the only adverse treatment effect) developed in treated and control birds but was more severe in the treatment group. Conclusions and Clinical Relevance —In African grey parrots, voriconazole has dose-dependent pharmacokinetics and may induce its own metabolism. Oral administration of 12 to 18 mg of voriconazole/kg twice daily is a rational starting dose for treatment of African grey parrots infected with Aspergillus or other fungal organisms that have a minimal inhibitory concentration for voriconazole ≤ 0.4 μg/mL. Higher doses may be needed to maintain plasma voriconazole concentrations during long-term treatment. Safety and efficacy of various voriconazole treatment regimens in this species require investigation. DA - 2008/1// PY - 2008/1// DO - 10.2460/ajvr.69.1.114 VL - 69 IS - 1 SP - 114-121 SN - 0002-9645 ER - TY - JOUR TI - Molecular epidemiologic investigation of Campylobacter coli in swine production systems, using multilocus sequence typing (vol 72, pg 5666, 2006) AU - Thakur, Siddhartha AU - Morrow, W. E. Morgan AU - Funk, Julie A. AU - Bahnson, Peter B. AU - Gebreyes, Wondwossen A. T2 - APPLIED AND ENVIRONMENTAL MICROBIOLOGY AB - Multilocus sequence typing of 151 Campylobacter coli isolates from swine reared in conventional (n = 74) and antimicrobial-free (n = 77) production systems revealed high genotypic diversity. Sequence type (ST) 1413 was predominant and observed among ciprofloxacin-resistant strains. We identified a C. coli ST 828 clonal complex consisting of isolates from both production systems. DA - 2008/1// PY - 2008/1// DO - 10.1128/aem.02326-07 VL - 74 IS - 1 SP - 342-342 SN - 0099-2240 ER - TY - JOUR TI - Oral Delivery of L-arginine Stimulates Prostaglandin-dependent Secretory Diarrhea in Cryptosporidium parvum–infected Neonatal Piglets AU - Gookin, Jody L AU - Foster, Derek M AU - Coccaro, Maria R AU - Stauffer, Stephen H T2 - Journal of Pediatric Gastroenterology and Nutrition AB - To determine if oral supplementation with L-arginine could augment nitric oxide (NO) synthesis and promote epithelial defense in neonatal piglets infected with Cryptosporidium parvum.Neonatal piglets were fed a liquid milk replacer and on day 3 of age infected or not with 10(8) C. parvum oocysts and the milk replacer supplemented with L-arginine or L-alanine. Milk consumption, body weight, fecal consistency, and oocyst excretion were recorded daily. On day 3 postinfection, piglets were euthanized and serum concentration of NO metabolites and histological severity of villous atrophy and epithelial infection were quantified. Sheets of ileal mucosa were mounted in Ussing chambers for measurement of barrier function (transepithelial resistance and permeability) and short-circuit current (an indirect measurement of Cl secretion in this tissue).C. parvum-infected piglets had large numbers of epithelial parasites, villous atrophy, decreased barrier function, severe watery diarrhea, and failure to gain weight. L-Arginine promoted synthesis of NO by infected piglets, which was unaccompanied by improvement in severity of infection but rather promoted epithelial chloride secretion and diarrhea. Epithelial secretion by infected mucosa from L-arginine-supplemented piglets was fully inhibited by the cyclooxygenase inhibitor indomethacin, indicating that prostaglandin synthesis was responsible for this effect.Results of these studies demonstrate that provision of additional NO substrate in the form of L-arginine incites prostaglandin-dependent secretory diarrhea and does not promote epithelial defense or barrier function of C. parvum-infected neonatal ileum. DA - 2008/2// PY - 2008/2// DO - 10.1097/MPG.0b013e31815c0480 VL - 46 IS - 2 SP - 139-146 J2 - Journal of Pediatric Gastroenterology and Nutrition LA - en OP - SN - 0277-2116 UR - http://dx.doi.org/10.1097/mpg.0b013e31815c0480 DB - Crossref KW - barrier function KW - enteral nutrition KW - epithelial defense KW - nitric oxide ER - TY - JOUR TI - Determination of minimum alveolar concentration of sevoflurane in juvenile swine AU - Moeser, A. J. AU - Blikslager, A. T. AU - Swanson, C. T2 - Research in Veterinary Science AB - Pigs are important animal models in veterinary and medical research and have been widely used in experiments requiring surgical anesthesia. Sevoflurane is an inhalant anesthetic with unique properties that make it an ideal anesthetic for mask induction and anesthesia maintenance. However, there are relatively few studies reporting the anesthetic requirements for sevoflurane in juvenile swine, an age group that is commonly used in research experiments. Therefore the objective of this study was to determine the Minimum Alveolar Concentration (MAC) for sevoflurane in juvenile swine. Sevoflurane anesthesia was induced in six Yorkshire-cross pigs of approximately 9 weeks-of-age and MAC for sevoflurane was determined. The sevoflurane MAC value was determined to be 3.5+/-0.1% which is notably higher than values reported in the literature for pigs. This discrepancy in MAC values may represent changes in anesthetic requirements between different age groups of pigs and differences in the type of stimulus used to determine MAC. DA - 2008/// PY - 2008/// DO - 10.1016/j.rvsc.2007.03.015 VL - 84 IS - 2 SP - 283-285 ER - TY - JOUR TI - Detection of two Bartonella tamiae-like sequences in Amblyomma americanum (Acari : Ixodidae) using 16S-23S intergenic spacer region-specific primers AU - Billeter, Sarah A. AU - Miller, Melissa K. AU - Breitschwerdt, Edward B. AU - Levy, Michael G. T2 - JOURNAL OF MEDICAL ENTOMOLOGY AB - Four hundred and sixty-six questing Amblyomma americanum (L.) (Acari: Ixodidae) from Carolina County, VA, and 98 questing A. americanum from Chatham County, NC, were screened by polymerase chain reaction (PCR) for the Bartonella 16S-23S intergenic spacer region. Two amplicons, approximately 270-280 bp, were detected in two ticks from Virginia. Based upon PCR and sequencing, an adult male and adult female tick harbored DNA sequences closely related to Bartonella tamiae (DQ395180). Bartonella DNA was not detected in A. americanum from North Carolina. Potential transmission of Bartonella spp. by A. americanum should be the focus of future experimental studies. DA - 2008/1// PY - 2008/1// DO - 10.1603/0022-2585(2008)45[176:DOTBTS]2.0.CO;2 VL - 45 IS - 1 SP - 176-179 SN - 0022-2585 KW - Bartonella KW - Amblyomma americanum KW - PCR ER - TY - JOUR TI - Arrginine activates intestinal p70(S6k) and protein Synthesis in piglet rotavivrus enteritis AU - Corl, Benjamin A. AU - Odle, Jack AU - Niu, Xiaomei AU - Moeser, Adam J. AU - Gatlin, Lori A. AU - Phillips, Oulayvanh T. AU - Blikslager, Anthony T. AU - Rhoads, J. Marc T2 - JOURNAL OF NUTRITION AB - We previously showed that phosphorylation of p70 S6 kinase (p70(S6k)) in the intestine is increased during viral enteritis. In this study, we hypothesized that during rotavirus infection, oral Arg, which stimulates p70(S6k) activation, will further stimulate intestinal protein synthesis and mucosal recovery, whereas the p70(S6k) inhibitor rapamycin (Rapa) will inhibit mucosal recovery. Newborn piglets were fed a standard milk replacer diet supplemented with Arg (0.4 g x kg(-1) x d(-1), twice daily by gavage), Rapa (2 mg x m(-2) x d(-1)), Arg + Rapa, or saline (controls). They were infected on d 6 of life with porcine rotavirus. Three days postinoculation, we measured the piglets' body weight, fecal rotavirus excretion, villus-crypt morphology, epithelial electrical resistance in Ussing chambers, and p70(S6k) activation by Western blotting and immunohistochemistry. We previously showed a 2-fold increase in jejunal protein synthesis during rotavirus diarrhea. In this experiment, Arg stimulated jejunal protein synthesis 1.3-fold above standard medium, and the Arg stimulation was partially inhibited by Rapa. Small bowel stimulation of p70(S6k) phosphorylation and p70(S6k) levels were inhibited >80% by Rapa. Immunohistochemistry revealed a major increase of p70(S6k) and ribosomal protein S6 phosphorylation in the crypt and lower villus of the infected piglets. However, in Arg-treated piglets, p70(S6k) activation occurred over the entire villus. Jejunal villi of the Rapa-treated group showed inactivation of p70(S6k) and a decrease in mucosal resistance (reflecting increased permeability), the latter of which was reversed by Arg. We conclude that, early in rotavirus enteritis, Arg has no impact on diarrhea but augments intestinal protein synthesis in part by p70(S6k) stimulation, while improving intestinal permeability via a mammalian target of rapamycin/p70(S6k)-independent mechanism. DA - 2008/1// PY - 2008/1// DO - 10.1093/jn/138.1.24 VL - 138 IS - 1 SP - 24-29 SN - 0022-3166 ER - TY - JOUR TI - Simplified hypoosmotic swelling testing (HOST) of fresh and frozen-thawed canine spermatozoa AU - Pinto, C. R. F. AU - Kozink, D. M. T2 - ANIMAL REPRODUCTION SCIENCE AB - The clinical use of the hypoosmotic swelling test (HOST) to identify spermatozoa with a functional intact membrane has been reported for humans and domestic species, including the dog. Currently, it is recommended that canine spermatozoa be incubated with the hypoosmotic solution for periods that range from 30 to 60 min. In an attempt to simplify the test, it was hypothesized that the degree of the hypoosmotic response at 1 min of incubation would not be different from the response documented at 60 min after incubation in the hypoosmotic solution at 37 degrees C. The hypoosmotic response of spermatozoa from 50 fresh and 16 frozen-thawed semen samples obtained from 22 adult dogs was recorded at 1 and 60 min of incubation. There were no significant differences between the hypoosmotic response recorded at 1 and 60 min for all evaluated semen samples (P>0.10). The hypoosmotic response recorded for canine spermatozoa from fresh semen samples were greater than that recorded for spermatozoa from frozen-thawed semen, both at 1 min (86.2% compared with 65.2%; P<0.001) and 60 min (85.6% compared with 61.8%; P<0.001). Based on the results of this study, it is recommended to decrease the incubation time of the HOST for canine spermatozoa to as short a period as 1 min. This incubation time should encourage the application of this relatively simple and inexpensive test of canine sperm membrane function in a clinical setting. DA - 2008/3/3/ PY - 2008/3/3/ DO - 10.1016/j.anireprosci.2007.07.005 VL - 104 IS - 2-4 SP - 450-455 SN - 1873-2232 KW - canine KW - sperm KW - sperm membrane KW - hypoosmotic swelling test ER - TY - JOUR TI - Selenium status and antibodies to selected pathogens in white-tailed deer (Odocoileus virginianus) in southern Minnesota AU - Wolf, Karen N. AU - DePerno, Christopher S. AU - Jenks, Jonathan A. AU - Stoskopf, Michael K. AU - Kennedy-Stoskopf, Suzanne AU - Swanson, Christopher C. AU - Brinkman, Todd J. AU - Osborn, Robert G. AU - Tardiff, Jeannine A. T2 - JOURNAL OF WILDLIFE DISEASES AB - To determine exposure to a variety of infectious diseases potentially important for native ungulates, livestock, and humans, serum samples from 114 (94 adults, 20 fawns) female white-tailed deer (Odocoileus virginianus) were collected during January 2000–03 from multiple locations in southeast (SE) and southwest (SW) Minnesota. Antibody prevalence was determined for the following pathogens: Mycobacterium avium subsp. paratuberculosis, Leptospira interrogans (six serovars), Anaplasma marginale, Borrelia burgdorferi, Brucella abortus, epizootic hemorrhagic disease virus, and bovine viral diarrhea virus (BVDV) types 1 and 2. Samples collected in 2001 were screened for antibodies against Anaplasma phagocytophilum, and whole blood was submitted for polymerase chain reaction (PCR) testing for A. phagocytophilum and B. burgdorferi. In addition, serum selenium concentrations were evaluated for samples collected during 2001– 03. Antibody prevalence and selenium concentration were compared by age-class and geographic region. Antibodies to all of the infectious agents except A. marginale and B. abortus were detected; when detected, antibody prevalence was highest in adults. Deer collected from SE Minnesota had a higher antibody prevalence to B. burgdorferi than SW deer. Blood culture and PCR results for A. phagocytophilum and B. burgdorferi were negative. Antibodies against BVDV (combined types 1 and 2) were more prevalent (χ2=3.617, P≤0.029) in deer collected in SW (41%) than in SE (25%) Minnesota. No statistically significant differences in serum selenium concentrations were detected when data were analyzed by age-class or by geographic location. DA - 2008/1// PY - 2008/1// DO - 10.7589/0090-3558-44.1.181 VL - 44 IS - 1 SP - 181-187 SN - 1943-3700 KW - infectious disease KW - Minnesota KW - Odocoileus virginianus KW - selenium KW - serology KW - white-tailed deer ER - TY - JOUR TI - Infectious diseases of dogs and cats on Isabela Island, Galapagos AU - Levy, J. K. AU - Crawford, P. C. AU - Lappin, M. R. AU - Dubovi, E. J. AU - Levy, M. G. AU - Alleman, R. AU - Tucker, S. J. AU - Clifford, E. L. T2 - JOURNAL OF VETERINARY INTERNAL MEDICINE AB - Background: Vaccination and importation of dogs and cats are prohibited in the Galapagos, resulting in a uniquely isolated population. The purpose of this study was to determine the prevalence of infectious diseases of dogs and cats that impact their health, could spill over to native wildlife, or sentinel diseases of concern to humans. Hypothesis: The isolation of dogs and cats in the Galapagos protects them from diseases common in mainland populations. Animals: Ninety‐five dogs and 52 cats presented during a neutering campaign. Methods: A prospective cross‐sectional study was performed. Blood was collected for serological and DNA evaluation of a panel of infectious diseases. Results: Antibodies against parvovirus (100%), parainfluenza virus (100%), adenovirus 1/2 (66–67%), and distemper virus (22%) were present in dogs. Dirofilaria immitis was also common in dogs (34%), with lower prevalences of Wolbachia pipiens (22%), Bartonella sp. (13%), Ehrlichia / Anaplasma spp. (1%), and Mycoplasma haemocanis (1%) observed. Antibodies against panleukopenia virus (67%), Toxoplasma gondii (63%), calicivirus (44%), and herpesvirus 1 (10%) were detected in cats. Feline leukemia virus antigen, feline immunodeficiency virus antibody, or coronavirus antibodies were not detected. Bartonella sp. (44%) infections were common in cats, but only one was infected with M. haemofelis . Conclusions and Clinical Importance: Despite their relative seclusion from the rest of the world, cats and dogs of Isabela were exposed to many pathogens found in mainland South America. Parasite prophylaxis, neutering, and strict enforcement of animal movement restrictions would control a majority of the diseases. In the absence of vaccination, a reservoir of susceptible animals remains vulnerable to new disease introductions. DA - 2008/// PY - 2008/// DO - 10.1111/j.1939-1676.2007.0034.x VL - 22 IS - 1 SP - 60-65 SN - 1939-1676 KW - bartonellosis KW - Dirofilaria immitis KW - distemper virus KW - parvovirus KW - toxoplasmosis KW - zoonoses ER - TY - JOUR TI - In vivo depletion of CD4(+)CD25(+) regulatory T cells in cats AU - Smithberg, S. Rochelle AU - Fogle, Jonathan E. AU - Mexas, Angela M. AU - Reckling, Stacie K. AU - Lankford, Susan M. AU - Tompkins, Mary B. AU - Dean, Gregg A. T2 - JOURNAL OF IMMUNOLOGICAL METHODS AB - To establish a characterized model of regulatory T cell (Treg) depletion in the cat we assessed the kinetics of depletion and rebound in peripheral and central lymphoid compartments after treatment with anti-CD25 antibody as determined by cell surface markers and FOXP3 mRNA expression. An 82% decrease in circulating CD4+CD25+ Tregs was observed by day 11 after treatment. CD4+CD25+ cells were also reduced in the thymus (69%), secondary lymphoid tissues (66%), and gut (67%). Although CD4+CD25+ cells rebound by day 35 post-treatment, FOXP3 levels remain depressed suggesting anti-CD25 antibody treatment has a sustainable diminutive effect on the Treg population. To determine whether CD25+ Treg depletion strategies also deplete activated CD25+ effector cells, cats were immunized with feline immunodeficiency virus (FIV) p24-GST recombinant protein, allowing them to develop a measurable memory response, prior to depletion with anti-CD25 antibody. Anti-FIV p24-GST effector cell activity in peripheral blood after depletion was sustained as determined by antigen-specific T cell proliferation and humoral responses against FIV p24-GST with an ELISA for antigen-specific feline IgG. Furthermore, development of an anti-mouse response in Treg-depleted cats was similar to control levels indicating the retained capacity to respond to a novel antigen. We conclude that despite alterations in CD25+ cell levels during depletion, the feline immune system remains functional. We demonstrate here a model for the study of disease pathogenesis in the context of reduced numbers of immunosuppressive CD4+CD25+ Tregs throughout the feline immune system. DA - 2008/1/1/ PY - 2008/1/1/ DO - 10.1016/j.jim.2007.09.015 VL - 329 IS - 1-2 SP - 81-91 SN - 0022-1759 KW - regulatory T cell KW - FOXP3 KW - feline immunodeficiency virus KW - monoclonal antibody ER - TY - JOUR TI - Exposure to phenobarbital in a foal after nursing a mare treated with phenobarbital AU - Wong, D. M. AU - Papich, M. G. AU - Davis, J. L. T2 - JOURNAL OF VETERINARY INTERNAL MEDICINE AB - A7-year-old, 672-kg, pregnant Thoroughbred mare presented with a 3-week history of premature lactation and lameness. The mare was a maiden, 336 days in gestation, and had been treated with 2 doses of flunixin meglumine (1 mg/kg, PO) the day before examination. Historically, the mare had experienced intermittent seizures while in race training. A complete diagnostic investigation to determine the cause of the seizures had been performed, but the origin of seizures remained undetermined. A weak positive Western blot analysis on cerebrospinal fluid for Sarcocystis neurona was reported at that time, and the mare was treated with ponazurila (5 mg/kg PO q24h) for 30 days.1 At examination, pertinent physical examination findings included mild tachycardia (56 beats/min), sensitivity to hoof testers involving all 4 feet, and the presence of thick, purulent exudate from the left mammary gland. Rectal palpation revealed anterior presentation of a live fetus, and transrectal and transabdominal ultrasonography demonstrated areas of uteroplacental thickening (14 mm; reference interval 7.14–11.98).2 While restrained in stocks, the mare demonstrated a mild partial seizure characterized by anxiousness, muscle fasciculations of the head and neck, and ataxia. Hematologic and serum biochemical abnormalities were limited to an increased plasma fibrinogen concentration (600 mg/dL; reference interval 100–400). Lateral radiography of the feet revealed mild soft tissue swelling at the coronary band in the right front foot, suggestive of laminitis with sinking of the 3rd phalanx within the hoof. Because of laminitis and suspicion of placentitis and mastitis, the mare was administered sulfamethoxazole-trimethoprimb (30 mg/kg PO q12h for 10 days) and phenylbutazonec (3 mg/kg PO q12h for 3 days, then 1.5 mg/kg PO q12h for 3 days), the left mammary gland was milked out q8h, and supportive pads were applied to all 4 feet. Two generalized seizures were observed 14 and 16 hours after initial examination, characterized by anxiousness, generalized muscle fasciculations most prominent around the head and neck, hyperesthesia, collapse, and recumbency. At this point, treatment with phenobarbitald (5.5 mg/kg PO q12h) was initiated. Seizure activity was not subsequently noted. Uncomplicated parturition of a 42.7 kg filly occurred 3 days after admission. The filly was lethargic and hypothermic (96.4°F; reference interval 99.5–101.3), had weak peripheral pulses, and maintained a low heart rate (52–56 bpm) for 45 minutes after birth. Heart rate and temperature progressively improved over the following 90 minutes. Blood and milk samples were collected from the mare and blood was collected from the foal immediately after parturition to measure phenobarbital concentrations. Phenobarbital concentration in the mare's serum was considered therapeutic (16.3 μg/mL; reference interval 15–40).3 The concentrations of phenobarbital in the mare's milk and foal's serum were 7.5 and 12.2 μg/mL, respectively. Initial treatment for the foal included intranasal oxygen (8 L/min for 1 hour) and intravenous administration of 2 L of hyperimmunized equine plasmae and ceftiofurf (10 mg/kg q6h × 7 days). A CBC, biochemistry profile, arterial blood gas analysis, and blood cultures were submitted immediately after parturition. No substantial clinicopathologic abnormalities were noted, and amikacing (25 mg/kg IV q24h × 7 days) was subsequently administered. Bacterial growth was not observed on blood culture. An 8 × 6 inch area of thickened irregular placenta was identified in the nonpregnant horn of the placenta. This section of placenta was characterized histologically by a fibrinonecrotic exudate covering the chorion and numerous filamentous Gram-positive rods. Blood and milk samples were collected from the mare and blood was collected from the foal at various times over the next 4 days of hospitalization to measure phenobarbital concentrations. The foal remained lethargic and demonstrated decreased activity until 48 hours of age when compared with a healthy foal of similar age. The foal was encouraged or assisted to stand for the 1st 48 hours to ensure adequate milk intake and progressively nursed more aggressively and regularly over time. The mare had no further signs of seizure activity, and the mare and foal were discharged 7 days after presentation. In total, the mare was administered phenobarbital twice daily for 6 days and then once daily for 3 days until treatement was discontinued. Blood and milk samples were collected from the mare and blood from the foal 3 and 7 days after discharge. The mare and foal were reported to be doing well 1 year after discharge with no evidence of seizures. The use of any drug in lactating mares can be controversial because the amount of drug the foal is exposed to is often unknown as there are very few scientific reports on this subject in the literature. Upon administration of medications to a lactating mare, a number of factors influence exposure of the neonate to the drug via milk. These include partitioning of drug from plasma into milk, drug concentration in milk, as well as the subsequent ingestion of the drug by the neonate.4–6 Because neonatal exposure to maternally administered drugs can have detrimental effects in the infant, mathematical calculations have been validated in humans to predict the neonate's exposure to a drug.7 The milk-to-plasma (M/P) ratio represents the ratio between the drug concentration in milk and that in maternal plasma/serum.5 This ratio provides an estimate of the amount of drug to which the suckling infant is exposed. Similarly, the formula used to predict the concentration of drug in milk provides an estimate of the amount of drug in milk, taking into consideration the amount of drug that is bound to protein. The calculation of exposure index (EI) was developed in people to link the M/P ratio, milk intake, and infant drug clearance to a time-averaged drug exposure level of the suckling infant.6 The EI attempts to account for differences in drug absorption and excretion in the suckling neonate when compared with the adult and is expressed as a relationship of the foal's exposure relative to the therapeutic dose of the mother.5 Blood samples were collected from the mare via the jugular vein and milk samples were collected from the right teat into sterile tubes immediately after parturition (time 0) and at 24, 48, 73, 74, 76, 78, 80, 83, 85, 86, 88, 90, 92, 95, 97, 167 (7 days), and 263 (11 days) hours after parturition. Phenobarbital had been administered to the mare for 2 days before parturition for a total of 4 doses. Treatment was continued 2 hours after parturition (ie, phenobarbital administered at time 2, 14, 26, 38, 50 hours after parturition). Blood from the foal was collected from a cephalic vein at parturition (time 0) and at 24, 48, 72, 74, 88, 167 (7 days), and 263 (11 days) hours after parturition. Serum was harvested and milk and serum samples were analyzed on the day of collection. Phenobarbital concentrations in serum and milk were measured by fluorescence polarization immunoassay (FPIA) on an automated chemistry analyzer.h The sensitivity of the assay, as reported by the manufacturer, was 1.1 μg/mL and the precision had a coefficient of variation less than 5%. Quality control (QC) samples were analyzed before the assay at low (15 μg/mL), medium (30 μg/mL), and high (50 μg/mL) concentrations to ensure that the assay performance was within QC ranges. Phenobarbital concentrations in the serum (▪) and milk (Δ) of the mare and the serum (•) of the foal over time after administration of 5.5 mg/kg PO q12h to the mare. Arrow indicates time of parturition, black arrowhead indicates time at which phenobarbital was switched to once daily administration, gray arrowhead indicates when phenobarbital administration was discontinued. Administration of phenobarbital in pregnant women is controversial because of its potential teratogenic effects.13 Furthermore, caution must be exercised in epileptic women receiving phenobarbital while breast feeding because this drug is transferred into breast milk and demonstrates a slow elimination by nursing infants.13,14 Phenobarbital is marginally excreted into human milk because milk pH is near the drug's pKa; therefore the drug is partially ionized and protein binding is moderate (51%).5 However, low clearance of the drug by the neonate can lead to accumulation of the drug and potentially result in blood concentrations that can produce adverse effects.5,14 Potential neonatal adverse effects include sedation and decreased suckling activity resulting from cumulative concentrations of phenobarbital in the neonate.5,14 Potential deleterious effects of phenobarbital in foals include respiratory depression, bradycardia, hypotension, hypothermia, and ataxia.11,15,16 The foal presented in this report demonstrated lethargy and decreased activity during the 1st 48 hours of life, which could have been attributed to several factors, including accumulation of phenobarbital within the fetus and continued exposure to phenobarbital through mare's milk. Phenobarbital administration to the mare was hastily discontinued because we felt that the risk of seizures was low postpartum. The etiology of the seizures in the mare remains uncertain. However, seizures in people can be precipitated by stress or pain, and factors such as strenuous training, impending parturition, and pain associated with laminitis and mastitis may have contributed to the mare's seizure activity.17 With correction or treatment of these factors, we believed the likelihood of seizures was outweighed by the detrimental effects of phenobarbital on the foal. The pharmacokinetic principles presented here demonstrate that it is possible to predict phenobarbital concentration in mare's milk from theoretical values. In the mare reported here, the concentration in milk (Fig 1) was slightly higher than, but similar to, the concentration predicted from equations using the mare M/P ratio. The milk concentration from predicted equations was 8.7 μg/mL at steady state whereas the actual milk concentration measured ranged from 9 to 11.2 μg/mL. Although our calculations predicted a close approximation of drug concentration in mare's milk after treatment with phenobarbital, the exposure to the foal was notably underestimated. The actual exposure of a nursing infant to a drug is influenced by the amount ingested and the ability of the neonate to clear the drug.4,6 The exposure index is expressed as the relationship of the foal's exposure relative to the exposure to a therapeutic dose to the mother. In this case it was predicted to be only 7.4%; however, the observed EI calculated from actual foal and mare plasma phenobarbital concentrations was 32%. This discrepancy may be a result of higher oral absorption or lower clearance of the drug in this foal than previously reported values.11 Realizing the limitations of extracting information from 1 horse, the case presented here reveals preliminary information with regard to phenobarbital disposition in a lactating mare and neonatal foal. It is apparent that phenobarbital was transferred to the fetus in utero and phenobarbital is excreted into the milk and absorbed by the suckling neonatal foal. In addition, methods shown here indicate that it may be possible to predict drug concentration in milk from the mare's serum concentration, but exposure to the foal requires a more accurate estimate of the foal's drug clearance and milk consumption. In clinical cases, it is encouraged to measure both maternal and neonatal serum concentrations of phenobarbital when a mare is being treated to monitor for possible over exposure to the foal and ensure therapeutic levels in the mare. aMarquis, Bayer HealthCare, Animal Health Division, Shawnee Mission, KS bSulfamethoxazole and trimethoprim, TEVA Pharmaceuticals, Sellersville, PA cPhenylbutazone, Phoenix Pharmaceutical, St Joseph, MO dPhenobarbital, Excellium Pharmaceutical, Fairfield, NJ eHyperimmunized Equine Plasma, MG Biologics, Ames, IA fNaxcel, Pharmacia & Upjohn, Kalamazoo, MI gAmikacin, Phoenix Pharmaceutical hAxSYM phenobarbital assay, Abbott Laboratories Diagnostic Division, Abbot Park, IL iWinNonlin version 5.1, Pharsight Corporation, Mountain View, CA The authors thank Ms Delta Dise, North Carolina State University, Clinical Pharmacology Laboratory, for her assistance with drug analysis for this study. DA - 2008/// PY - 2008/// DO - 10.1111/j.1939-1676.2007.0036.x VL - 22 IS - 1 SP - 227-230 SN - 1939-1676 ER - TY - JOUR TI - Concurrent hepatic copper toxicosis and Fanconi's syndrome in a dog AU - Hill, T. L. AU - Breitschwerdt, E. B. AU - Cecere, T. AU - Vaden, S. T2 - JOURNAL OF VETERINARY INTERNAL MEDICINE AB - A3-year-old male castrated West Highland White Terrier was presented to the Veterinary Teaching Hospital at North Carolina State University with a 1-week history of intermittent vomiting, polyuria and polydypsia, progressive anorexia, and lethargy. No treatment was administered by the primary veterinarian before referral. The dog was adopted as a puppy and had lived exclusively in North Carolina. The dog received no other medications and had no previous illnesses, except for a transient decrease in appetite 1 year earlier. On physical examination, the dog weighed 8.2 kg, had a body condition score of 6/9, but was approximately 5% dehydrated. Hematologic abnormalities included neutrophilia (14,293 cells/μL, reference range 2,529–12,876 cells/μL) with a left shift (642 band neutrophils/μL) and immature granulocytes (161/μL). Biochemical abnormalities included increased ALT activity (456 U/L; reference range, 16–73 U/L), hyperbilirubinemia (0.3 mg/dL; reference range, 0–0.2 mg/dL), hypokalemia (3.8 mEq/L; reference range, 3.9–5.2 mEq/L), hyperchloremia (121 mEq/L; reference range, 104–117 mEq/L), and increased lipase activity (715 U/L; reference range, 22–216 U/L). Urinalysis findings included a specific gravity of 1.022, pH of 8, 2+ proteinuria, 2+ ketones, 2+ blood, 4+ glucose, 0–2 coarse granular casts/hpf, 0–5 white cells/hpf, and 5–10 red cells/hpf. The urine protein : creatinine ratio was 1.7. A urine sample submitted for bacterial culture yielded no growth. There was mild metabolic acidemia (blood pH 7.26; reference range, 7.36–7.47), bicarbonate 16.2 mEq/L (reference range, 19.8–26.2 mEq/L), and PCO2 36 mmHg (reference range, 30–40 mmHg), on a venous blood sample. Abdominal ultrasonographic findings included periportal lymph node enlargement, microhepatica, and mildly hyperechoic kidneys bilaterally. Fasting and postprandial serum bile acid concentrations were within reference range. The dog was treated with an IV infusion of lactated Ringer's solution containing 30 mEq/L KCl for dehydration, hypokalemia, and metabolic acidosis. Within 48 hours of hospitalization, sodium bicarbonate at 3 mEq/kg/d was administered as a continuous rate infusion to correct the persistent metabolic acidosis. In addition, the dog was treated with a metoclopramide constant rate infusion of 1 mg/kg/d and famotidine 0.5 mg/kg IV q12h. During the 1st 48 hours of hospitalization, the dog remained anorexic and frequently vomited bile; dolasetron 0.6 mg/kg IV q24h and sucralfate 500 mg PO q8h were administered. The dog was supplemented nutritionally by nasoesophageal feeding a liquid diet (Perativea), the continuous rate of which was adjusted to minimize vomiting. Serial assessment of urine dipstick tests and serum glucose measurements indicated that the dog was persistently ketonuric and glucosuric, with normal serum glucose concentrations. Despite a metabolic acidosis, urine pH ranged from 7.0 to 8.0. The findings of proteinuria, glucosuria with normoglycemia, and hyperchloremic metabolic acidosis with alkaline urine supported a diagnosis of Fanconi's syndrome. Results of a urine metabolic profile, performed at the University of Pennsylvania, also were consistent with Fanconi's syndrome with severe generalized amino aciduria and marked glucosuria. By the 4th day of hospitalization, the dog became febrile (103.6°F). An abdominal ultrasound examination indicated mild thickening of the gallbladder wall. Antibiotic therapy was initiated with ampicillin and sulbactam (Unasyn,b 30 mg/kg IV q8h); the fever resolved within 36 hours. Because of the dog's refractory vomiting and unknown underlying hepatic pathology, an exploratory laparotomy was performed on the 5th day of hospitalization. Biopsy specimens were taken of the liver, stomach, duodenum, jejunum, and left kidney. The gallbladder was aspirated and 1 aliquot of bile was submitted for cytologic analysis; another aliquot was submitted for aerobic microbial culture. Gastrostomy and jejunostomy tubes were placed. Recovery from surgery was uneventful. Perativea liquid diet was administered continuously through the jejunostomy tube throughout the remainder of hospitalization without incident. The volume administered was gradually increased to basal energy requirements. Ketonuria resolved within 24 hours after feeding basal energy requirements. Dexamethasone (0.08 mg/kg IV q24h), s-adenosylmethionine (22 mg/kg PO q24h), and ursodeoxycholic acid (15 mg/kg PO q24h) were administered on day 5 postoperatively. Within 24 hours, steady improvement was observed; the vomiting ceased, the dog was more active and alert, and began eating. IV bicarbonate supplementation was necessary to maintain a near normal serum pH. The bile fluid was cytologically unremarkable and bacterial growth was not observed. Lymphofollicular gastritis, which was attributed to refractory vomiting, and mild lymphocytic enteritis were observed histologically on full-thickness stomach and intestinal biopsy specimens. Centrilobular pyogranulomatous hepatitis, characterized by infiltrates of neutrophils, macrophages, and fewer lymphocytes and plasma cells associated with single cell hepatocyte necrosis, was present in liver wedge biopsy specimens. Abundant copper accumulation was present within centrilobular hepatocytes and macrophages (Fig 1). Copper was quantified at 1186 ppm dry weight. Histopathologic abnormalities in the kidney included diffuse mild to moderate tubular atrophy, multifocal tubular epithelial vacuolation, and tubular regeneration. Additionally, there was evidence of abnormal copper accumulation within vacuolated renal tubular epithelium (Fig 2). Liver, dog. (A) Centrilobular zone with individual hepatocellular necrosis (arrow) and infiltrates of macrophages, neutrophils, lymphocytes, and plasma cells. H&E stain. Scale bar = 50 μm. (B) Same section as A with numerous well-demarcated, red–brown copper-containing granules present in the cytoplasm of macrophages and hepatocytes. Rhodanine stain. Scale bar = 50 μm. Kidney, dog. (A) Tubules lined by plump vacuolated epithelial cells (arrow) or mildly atrophied epithelium (arrowhead). H&E stain. Scale bar = 20 μm. (B) Renal tubular epithelial cells containing multiple cytoplasmic well-demarcated, red–brown copper-containing granules. Rhodanine stain. Scale bar = 20 μm. The dog gradually was transitioned to oral medications administered through the gastrotomy tube, including 971 mg of bicarbonate q12h, s-adenosylmethionine, ursodeoxycholic acid, prednisone 0.6 mg/kg q12h that was tapered over the next 4 weeks, famotidine 0.6 mg/kg q12h for 2 weeks, amoxicillin–clavulanic acid 12 mg/kg q12h for 2 weeks, ciprofloxacin 8 mg/kg q12h for 2 weeks, and Marinc 1 medium dog tablet daily for 2 months. d-Penicillamine was given at a dosage of 10 mg/kg PO q12h for 3 months, with no reported adverse effects. After 2 weeks of treatment, the dog was no longer proteinuric, glucosuric, or ketonuric. Venous pH was 7.36 and bicarbonate was 26.6 mEq/L. ALT activity had decreased from 456 to 81 U/L. Ursodeoxycholic acid, s-adenosylmethionine, Marin, bicarbonate, and d-penicillamine therapy were continued. Monthly venous blood gas analysis disclosed normal pH while receiving oral bicarbonate supplementation. At the 3-month recheck examination, the bicarbonate dosage was reduced by half and d-penicillamine was discontinued. Repeat biopsies of the liver and kidney were declined by the owner. Zinc acetate was prescribed (10 mg/kg orally q12h); s-adenosylmethionine, ursodeoxycholic acid, and Marin were continued. Three weeks after decreasing the oral bicarbonate supplementation, venous pH was 7.4 and bicarbonate was 27.9 mEq/L and bicarbonate supplementation was discontinued, but zinc acetate supplementation was continued indefinitely. Thirteen months after initial diagnosis and treatment, the dog was doing well with no clinical signs reported. Copper storage diseases (CSDs) have been described in several breeds, including the Bedlington Terrier, West Highland White Terrier, Skye Terrier, Doberman Pinscher, Labrador Retriever, and Dalmatian, as well as other species, including humans, rats, and sheep. With CSD, copper accumulates in the liver, leading to hepatitis and eventually cirrhosis of the hepatic parenchyma.1 As yet, the genetic basis of CSD has been elucidated only in the Bedlington Terrier, where it is related to a defect in the MURR-1 gene.2 CSD in the Bedlington Terrier results in copper accumulation in the liver as well as renal cortical tissue in some patients.3 We describe a West Highland White Terrier with CSDand concurrent Fanconi's syndrome, which resolved after copper chelation therapy. A genetic mutation that causes abnormal copper accumulation has yet to be identified in West Highland White Terrier. Unlike the Bedlington Terrier, there does not seem to be a correlation between age and hepatic copper concentration in the West Highland White Terrier. In addition, the total hepatic copper concentration is lower in the majority of affected West Highland White Terriers; most West Highland White Terriers have copper concentrations <1,500 ppm dry weight.4, 5 Copper concentrations up to 500 ppm dry weight may occur in normal dogs.6 Thornburg describes the histopathologic lesions of copper toxicosis in West Highland White Terriers as being characterized by multifocal centrilobular hepatitis and cirrhosis. In this dog, the findings were consistent with previous reports, including multifocal centrilobular hepatitis and a copper concentration of 1,186 ppm. It is unclear whether copper toxicosis was the direct cause of Fanconi's syndrome in this dog, because several medical therapies, including antibiotics, steroids, and penicillamine, may have contributed to resolution of Fanconi's syndrome regardless of treating copper toxicosis. The presence of copper in the renal tubules and resolution of Fanconi's syndrome after copper chelation therapy suggest that copper toxicosis may be a cause of Fanconi's syndrome in dogs. In other reports, dogs with copper toxicosis had abnormalities indicative of proximal tubular dysfunction.7, 8 In a study of 10 Dalmatians with copper toxicosis, 3 dogs had proteinuria without pyuria, 2 had glucosuria with normoglycemia, and 1 had renal tubular necrosis with granular casts.7 In a separate report, a 1.5-year-old Dalmatian had copper toxicosis with a positive metabolic screen for Fanconi's syndrome.8 This dog was euthanized because the dog continued to decline clinically. In the dog reported here, glucosuria, metabolic acidosis, amino aciduria, and copper accumulation in the renal tubules were documented with resolution of Fanconi's syndrome after copper chelation therapy. Wilson's disease is a CSD in humans in which copper accumulates to toxic concentrations in the liver and secondarily in the central nervous system and kidneys because of a mutation in the ATP7B gene, which normally allows for copper excretion into the bile and for production of ceruloplasmin. As a result, patients with Wilson's disease have copper accumulation in the liver and in other tissues, including the brain, kidney, red blood cells, and eye. Neurologic signs develop including speech disorders and dysphagia, abnormal and uncoordinated gait, and tremors.9 Renal complications, including urolithiasis and Fanconi's syndrome, have been reported in human patients with Wilson's disease.10, 11 As is reported in association with Wilson's disease in human patients, this dog may represent a subset of patients with copper storage disease that have concurrent renal tubular dysfunction in association with copper accumulation in the proximal tubular epithelium. Several types of proximal renal tubular dysfunction have been described in association with Wilson's disease in humans: failure of renal acidification, amino aciduria, glucosuria, and phosphaturia.11-15 Resolution of proximal tubular dysfunction also may occur after treatment for copper toxicosis with penicillamine.11, 13-15 A renal biopsy in 1 patient, with prior documentation of renal tubular copper accumulation, demonstrated normal proximal tubular ultrastructure 2 years after initiation of penicillamine therapy. Penicillamine therapy was discontinued because of adverse effects, including glomerulonephritis and systemic lupus erythematosus. Eighteen months later, the patient again showed signs of Wilson's disease, and a renal biopsy was repeated. Electron-dense bodies, consistent with copper-bound metalloprotein, were evident in the subapical cytoplasm of the tubular cells, although copper quantification was not performed to confirm increased copper concentrations in the kidney.11 The effect of copper loading has been examined in the rat kidney as a model for copper toxicosis in humans and other species. Rats supplemented with excessive copper, either by injection or by dietary supplementation, develop copper staining in the liver and in the proximal convoluted tubular epithelium.16-18 Haywood demonstrated an increase in copper content of the kidney in rats fed excess copper as well as copper staining granules confined to the proximal tubule. There also were degenerative changes of the tubular cells as well as copper-staining debris in the tubular lumen, suggesting active exocytosis of copper-bound metallothionein.17 A later report of copper loading in the rat kidney described increased copper in renal tubular lysosomes. With time and increased copper concentrations, there was progressive nuclear degeneration in proximal tubular cells and disruption of the mitochondrial membrane.18 These findings are consistent with the observation that copper acts as a prooxidant, disrupting cell membranes and damaging DNA. Eventually, the rats extruded copper-stained lysosomes and copper-laden pinocytotic vesicles into the tubular lumen. After this time point, the copper concentration in the kidney began to decrease and the tubular epithelium recovered to nearly normal. The localization of copper to the renal tubular epithelium, later exocytosis of copper-bound organelles, and recovery of the tubular epithelium suggest a mechanism by which the rat seems able to cope with increased dietary copper intake. Copper also may alter the function of Na-K-ATPase in the proximal tubular epithelium. In vitro, for both rat kidney tissue homogenate and rat synaptic plasma membrane, copper has an inhibitory effect on the function of this important enzyme in a concentration-dependent manner.19, 20 As copper stores accumulate in the canine liver, the kidney may attempt exocytosis of excess copper as occurs in the copper-loaded rat. Exocytosis of copper-bound organelles in the canine and human kidney may be less effective than in the rat, because tubular debris is not described in any published reports of copper-stained kidneys from rats. Because copper accumulates in the kidney as well as the liver, it then may have several effects that ultimately lead to proximal tubular dysfunction and Fanconi's syndrome: necrosis and apoptosis of epithelial cells because copper acts as a pro-oxidant, disrupting mitochondrial membranes and DNA, inducing inflammation that may affect epithelial function, and inhibiting the function of Na-K-ATPase in a concentration-dependent manner that would alter transport mechanisms in the proximal tubule. These effects all may lead to decreased reabsorption of glucose, amino acids, phosphate, and bicarbonate from the tubular lumen. These may be the mechanisms by which copper toxicosis in this West Highland White Terrier resulted in proximal tubular dysfunction characterized as Fanconi's syndrome. Few controlled studies are available in human and canine medicine that describe renal pathology with copper toxicosis or the effect of copper chelation therapy. Additional study is needed to definitively confirm a link between Fanconi's syndrome and copper storage disease in dogs as suggested by this dog, and to elucidate the nature of that association. In dogs with suspected copper storage disease and evidence of tubular dysfunction that are undergoing liver biopsy, it may be warranted to perform a renal biopsy to allow for histopathologic examination of the renal parenchyma, as well as renal copper quantification, which was not performed here. Sequential urine metabolic screening or urinalyses with protein quantification may be a useful diagnostic and therapeutic monitoring tool in those patients with evidence of tubular dysfunction. Additionally, liver and kidney biopsies after 3 months of d-penicillamine therapy may have been informative to document the histological response to treatment. The authors gratefully acknowledge the assistance of the veterinarians involved in the treatment and referral of the dog. Our appreciation is extended to Dr Herman Jeffers, Dr Karyn Harrell, and Dr Sally Bissett. aPerative Specialized Nutrition, Abbot Laboratories, Ross Products Division, Columbus, OH bUnasyn, Pfizer Roerig, Pfizer Inc, New York cMarin, Nutramax Laboratories Inc, Edgewood, MD DA - 2008/// PY - 2008/// DO - 10.1111/j.1939-1676.2007.0040.x VL - 22 IS - 1 SP - 219-222 SN - 0891-6640 ER - TY - JOUR TI - Bartonella DNA in the blood and lymph nodes of golden retrievers with lymphoma and in healthy controls AU - Duncan, A. W. AU - Marr, H. S. AU - Birkenheuer, A. J. AU - Maggi, R. G. AU - Williams, L. E. AU - Correa, M. T. AU - Breitschwerdt, E. B. T2 - JOURNAL OF VETERINARY INTERNAL MEDICINE AB - Background: Although lymphoma is the most common neoplastic process reported in dogs, its precise etiology is unknown. Golden Retrievers are more likely to develop lymphoma, suggesting a breed predisposition; however, other factors, including environment, immunity, and infection, are likely contributors to oncogenesis. Hypothesis: We hypothesized that the development of lymphoma in Golden Retrievers may be associated with vector‐borne infections, specifically Bartonella , Anaplasma , or Ehrlichia species infections. Animals: Golden Retrievers with lymphoma and healthy Golden Retrievers from across the United States were recruited for study participation. Methods: A matched, case‐control study was performed to determine the association of lymphoma and the presence of Bartonella, Anaplasma , and Ehrlichia species in serum, blood, and lymph node aspirates. Results: Using PCR analyses and DNA sequencing, single and coinfections with Bartonella henselae , Bartonella elizabethae, Bartonella quintana , and/or Bartonella vinsonii ( berkhoffii ) were detected in the blood and lymph node aspirates of Golden Retrievers with lymphoma (5/28 dogs, 18%) and in healthy Golden Retrievers (10/56 dogs, 18%); no Anaplasma or Ehrlichia DNA was detected in any dog. When compared with dogs with lymphoma, a higher ( P <.001) proportion of healthy Golden Retrievers were receiving monthly acaricide treatments (2.6 times higher). Conclusions and Clinical Importance: Bartonella DNA can be detected in blood and lymph nodes; importantly, in this report, Bartonella was detected in the same proportion of clinically healthy dogs and dogs with lymphoma. Longitudinal studies should be conducted to determine the mode of transmission of Bartonella in dogs, whether lymphatic infection is persistent, or whether these bacteria may contribute to the development of lymphoma. DA - 2008/// PY - 2008/// DO - 10.1111/j.1939-1676.2007.0018.x VL - 22 IS - 1 SP - 89-95 SN - 0891-6640 KW - bacteria KW - cancer KW - dog KW - tick KW - vector-borne ER -