@article{brooks_landau_everitt_brown_grady_waskowicz_bass_d'angelo_asfaw_williams_et al._2018, title={Long-term complications of glycogen storage disease type Ia in the canine model treated with gene replacement therapy}, volume={41}, ISSN={["1573-2665"]}, DOI={10.1007/s10545-018-0223-y}, abstractNote={Background Glycogen storage disease type Ia (GSD Ia) in dogs closely resembles human GSD Ia. Untreated patients with GSD Ia develop complications associated with glucose-6-phosphatase (G6Pase) deficiency. Survival of human patients on intensive nutritional management has improved; however, long-term complications persist including renal failure, nephrolithiasis, hepatocellular adenomas (HCA), and a high risk for hepatocellular carcinoma (HCC). Affected dogs fail to thrive with dietary therapy alone. Treatment with gene replacement therapy using adeno-associated viral vectors (AAV) expressing G6Pase has greatly prolonged life and prevented hypoglycemia in affected dogs. However, long-term complications have not been described to date. Methods Five GSD Ia-affected dogs treated with AAV-G6Pase were evaluated. Dogs were euthanized due to reaching humane endpoints related to liver and/or kidney involvement, at 4 to 8 years of life. Necropsies were performed and tissues were analyzed. Results Four dogs had liver tumors consistent with HCA and HCC. Three dogs developed renal failure, but all dogs exhibited progressive kidney disease histologically. Urolithiasis was detected in two dogs; uroliths were composed of calcium oxalate and calcium phosphate. One affected and one carrier dog had polycystic ovarian disease. Bone mineral density was not significantly affected. Conclusions Here, we show that the canine GSD Ia model demonstrates similar long-term complications as GSD Ia patients in spite of gene replacement therapy. Further development of gene therapy is needed to develop a more effective treatment to prevent long-term complications of GSD Ia.}, number={6}, journal={JOURNAL OF INHERITED METABOLIC DISEASE}, author={Brooks, Elizabeth D. and Landau, Dustin J. and Everitt, Jeffrey I. and Brown, Talmage T. and Grady, Kylie M. and Waskowicz, Lauren and Bass, Cameron R. and D'Angelo, John and Asfaw, Yohannes G. and Williams, Kyha and et al.}, year={2018}, month={Dec}, pages={965–976} }
 @article{demaster_luo_curtis_williams_landau_drake_kozink_bird_crane_sun_et al._2012, title={Long-Term Efficacy Following Readministration of an Adeno-Associated Virus Vector in Dogs with Glycogen Storage Disease Type Ia}, volume={23}, ISSN={["1557-7422"]}, DOI={10.1089/hum.2011.106}, abstractNote={Glycogen storage disease type Ia (GSD-Ia) is the inherited deficiency of glucose-6-phosphatase (G6Pase), primarily found in liver and kidney, which causes life-threatening hypoglycemia. Dogs with GSD-Ia were treated with double-stranded adeno-associated virus (AAV) vectors encoding human G6Pase. Administration of an AAV9 pseudotyped (AAV2/9) vector to seven consecutive GSD-Ia neonates prevented hypoglycemia during fasting for up to 8 hr; however, efficacy eventually waned between 2 and 30 months of age, and readministration of a new pseudotype was eventually required to maintain control of hypoglycemia. Three of these dogs succumbed to acute hypoglycemia between 7 and 9 weeks of age; however, this demise could have been prevented by earlier readministration an AAV vector, as demonstrated by successful prevention of mortality of three dogs treated earlier in life. Over the course of this study, six out of nine dogs survived after readministration of an AAV vector. Of these, each dog required readministration on average every 9 months. However, two were not retreated until >34 months of age, while one with preexisting antibodies was re-treated three times in 10 months. Glycogen content was normalized in the liver following vector administration, and G6Pase activity was increased in the liver of vector-treated dogs in comparison with GSD-Ia dogs that received only with dietary treatment. G6Pase activity reached approximately 40% of normal in two female dogs following AAV2/9 vector administration. Elevated aspartate transaminase in absence of inflammation indicated that hepatocellular turnover in the liver might drive the loss of vector genomes. Survival was prolonged for up to 60 months in dogs treated by readministration, and all dogs treated by readministration continue to thrive despite the demonstrated risk for recurrent hypoglycemia and mortality from waning efficacy of the AAV2/9 vector. These preclinical data support the further translation of AAV vector–mediated gene therapy in GSD-Ia. Demaster and colleagues report preclinical results of treating dogs with glycogen storage disease type Ia (GSD-Ia) with double-stranded adeno-associated viral vector type 9 (AAV2/9) encoding human glucose-6-phosphatase (G6Pase). Vector treatment was able to prevent hypoglycemia and led to normalized liver glycogen levels and increased G6Pase activity, but carefully timed readministration with a different pseudotype was required to prevent the therapeutic effect from waning. Survival was prolonged for up to 60 months in dogs treated by readministration, with all dogs continuing to thrive despite the demonstrated risk for recurrent hypoglycemia and mortality from waning vector efficacy.}, number={4}, journal={HUMAN GENE THERAPY}, author={Demaster, Amanda and Luo, Xiaoyan and Curtis, Sarah and Williams, Kyha D. and Landau, Dustin J. and Drake, Elizabeth J. and Kozink, Daniel M. and Bird, Andrew and Crane, Bayley and Sun, Francis and et al.}, year={2012}, month={Apr}, pages={407–418} }
 @article{crane_luo_demaster_williams_kozink_zhang_brown_pinto_oka_sun_et al._2012, title={Rescue administration of a helper-dependent adenovirus vector with long-term efficacy in dogs with glycogen storage disease type Ia}, volume={19}, ISSN={["0969-7128"]}, DOI={10.1038/gt.2011.86}, abstractNote={Glycogen storage disease type Ia (GSD-Ia) stems from glucose-6-phosphatase (G6Pase) deficiency and causes hypoglycemia, hepatomegaly, hypercholesterolemia and lactic acidemia. Three dogs with GSD-Ia were initially treated with a helper-dependent adenovirus encoding a human G6Pase transgene (HDAd-cG6Pase serotype 5) on postnatal day 3. Unlike untreated dogs with GSD-Ia, all three dogs initially maintained normal blood glucose levels. After 6–22 months, vector-treated dogs developed hypoglycemia, anorexia and lethargy, suggesting that the HDAd-cG6Pase serotype 5 vector had lost efficacy. Liver biopsies collected at this time revealed significantly elevated hepatic G6Pase activity and reduced glycogen content, when compared with affected dogs treated only by frequent feeding. Subsequently, the HDAd-cG6Pase serotype 2 vector was administered to two dogs, and hypoglycemia was reversed; however, renal dysfunction and recurrent hypoglycemia complicated their management. Administration of a serotype 2 HDAd vector prolonged survival in one GSD-Ia dog to 12 months of age and 36 months of age in the other, but the persistence of long-term complications limited HDAd vectors in the canine model for GSD-Ia.}, number={4}, journal={GENE THERAPY}, author={Crane, B. and Luo, X. and Demaster, A. and Williams, K. D. and Kozink, D. M. and Zhang, P. and Brown, T. T. and Pinto, C. R. and Oka, K. and Sun, F. and et al.}, year={2012}, month={Apr}, pages={443–452} }
 @article{fry_brown_lloyd_hansen_legleiter_robarge_spears_2011, title={Effect of dietary boron on physiological responses in growing steers inoculated with bovine herpesvirus type-1}, volume={90}, ISSN={["1532-2661"]}, DOI={10.1016/j.rvsc.2010.04.016}, abstractNote={Thirty-six Angus and Angus × Simmental steers were fed one of three dietary treatments; (1) control (no supplemental B), (2) 5 mg supplemental B/kg, and (3) 15 mg supplemental B/kg for 47 days to determine the effects of dietary boron (B) on disease resistance following an inoculation with bovine herpesvirus type-1 (BHV-1). On day 34 of the study steers were inoculated intranasally with BHV-1. Rectal temperatures began to elevate at day 2, and plasma tumor necrosis factor-α concentrations increased (P < 0.05) by day 2 following BHV-1 inoculation. Plasma acute phase proteins were increased (P < 0.01) while plasma interferon-γ was decreased (P < 0.05) by day 4 post-inoculation. Supplementation of B increased (P < 0.001) plasma B concentrations in a dose-responsive manner. However, dietary B did not affect the duration and severity of clinical signs of BHV-1 and had minimal effects on plasma acute phase proteins and cytokines.}, number={1}, journal={RESEARCH IN VETERINARY SCIENCE}, author={Fry, R. S. and Brown, T. T., Jr. and Lloyd, K. E. and Hansen, S. L. and Legleiter, L. R. and Robarge, W. P. and Spears, J. W.}, year={2011}, month={Feb}, pages={78–83} }
 @article{luo_hall_li_bird_lavin_winn_kemper_brown_koeberl_2011, title={Hepatorenal Correction in Murine Glycogen Storage Disease Type I With a Double-stranded Adeno-associated Virus Vector}, volume={19}, ISSN={["1525-0016"]}, DOI={10.1038/mt.2011.126}, abstractNote={Glycogen storage disease type Ia (GSD-Ia) is caused by the deficiency of glucose-6-phosphatase (G6Pase). Long-term complications of GSD-Ia include life-threatening hypoglycemia and proteinuria progressing to renal failure. A double-stranded (ds) adeno-associated virus serotype 2 (AAV2) vector encoding human G6Pase was pseudotyped with four serotypes, AAV2, AAV7, AAV8, and AAV9, and we evaluated efficacy in 12-day-old G6pase (−/−) mice. Hypoglycemia during fasting (plasma glucose <100 mg/dl) was prevented for >6 months by the dsAAV2/7, dsAAV2/8, and dsAAV2/9 vectors. Prolonged fasting for 8 hours revealed normalization of blood glucose following dsAAV2/9 vector administration at the higher dose. The glycogen content of kidney was reduced by >65% with both the dsAAV2/7 and dsAAV2/9 vectors, and renal glycogen content was stably reduced between 7 and 12 months of age for the dsAAV2/9 vector-treated mice. Every vector-treated group had significantly reduced glycogen content in the liver, in comparison with untreated G6pase (−/−) mice. G6Pase was expressed in many renal epithelial cells of with the dsAAV2/9 vector for up to 12 months. Albuminuria and renal fibrosis were reduced by the dsAAV2/9 vector. Hepatorenal correction in G6pase (−/−) mice demonstrates the potential of AAV vectors for the correction of inherited diseases of metabolism. Glycogen storage disease type Ia (GSD-Ia) is caused by the deficiency of glucose-6-phosphatase (G6Pase). Long-term complications of GSD-Ia include life-threatening hypoglycemia and proteinuria progressing to renal failure. A double-stranded (ds) adeno-associated virus serotype 2 (AAV2) vector encoding human G6Pase was pseudotyped with four serotypes, AAV2, AAV7, AAV8, and AAV9, and we evaluated efficacy in 12-day-old G6pase (−/−) mice. Hypoglycemia during fasting (plasma glucose <100 mg/dl) was prevented for >6 months by the dsAAV2/7, dsAAV2/8, and dsAAV2/9 vectors. Prolonged fasting for 8 hours revealed normalization of blood glucose following dsAAV2/9 vector administration at the higher dose. The glycogen content of kidney was reduced by >65% with both the dsAAV2/7 and dsAAV2/9 vectors, and renal glycogen content was stably reduced between 7 and 12 months of age for the dsAAV2/9 vector-treated mice. Every vector-treated group had significantly reduced glycogen content in the liver, in comparison with untreated G6pase (−/−) mice. G6Pase was expressed in many renal epithelial cells of with the dsAAV2/9 vector for up to 12 months. Albuminuria and renal fibrosis were reduced by the dsAAV2/9 vector. Hepatorenal correction in G6pase (−/−) mice demonstrates the potential of AAV vectors for the correction of inherited diseases of metabolism.}, number={11}, journal={MOLECULAR THERAPY}, author={Luo, Xiaoyan and Hall, Gentzon and Li, Songtao and Bird, Andrew and Lavin, Peter J. and Winn, Michelle P. and Kemper, Alex R. and Brown, Talmage T. and Koeberl, Dwight D.}, year={2011}, month={Nov}, pages={1961–1970} }
 @article{koeberl_pinto_brown_chen_2009, title={Gene Therapy for Inherited Metabolic Disorders in Companion Animals}, volume={50}, ISSN={["1084-2020"]}, DOI={10.1093/ilar.50.2.122}, abstractNote={Scientists first described inborn errors of metabolism, also termed inherited disorders of metabolism, early in the 20th century and since then have determined the biochemical and genetic bases of a great number of these disorders both in humans and in an increasing number of companion animals. The availability of metabolic screening tests has advanced the biochemical and genetic characterization in affected breeds of companion animals of inherited metabolic disorders involving amino acid, carbohydrate, fatty acid, and metal metabolism. Advances in gene therapy have led to the development of new treatments for inherited disorders of metabolism, and animal models have played a critical role in this research. For example, glycogen storage disease type Ia in dogs was highly responsive to adeno-associated viral vector–mediated gene therapy, which prolonged survival and for more than a year prevented hypoglycemia during fasting. Gene therapy for other glycogen storage diseases and metabolic disorders will also be feasible. The establishment of a breeding colony and the ability to sustain affected animals are critical steps toward evaluating the safety and efficacy of gene therapy with clinically relevant endpoints. The further development of gene therapy for inherited disorders of metabolism could lead to curative therapy for affected humans and animals alike.}, number={2}, journal={ILAR JOURNAL}, author={Koeberl, Dwight D. and Pinto, Carlos and Brown, Talmage and Chen, Y. T.}, year={2009}, pages={122–127} }
 @article{hill_hopkins_davidson_bolt_diaz_brownie_brown_huntington_whitlow_2009, title={The addition of cottonseed hulls to the starter and supplementation of live yeast or mannanoligosaccharide in the milk for young calves}, volume={92}, ISSN={["1525-3198"]}, DOI={10.3168/jds.2008-1320}, abstractNote={The objectives of this study were to investigate the effects of the addition of cottonseed hulls (CSH) to the starter and the supplementation of live yeast product (YST) or mannanoligosaccharide product (MOS) to milk, on growth, intake, rumen development, and health parameters in young calves. Holstein (n = 116) and Jersey (n = 46) bull (n = 74) and heifer (n = 88) calves were assigned randomly within sex at birth to treatments. All calves were fed 3.8 L of colostrum daily for the first 2 d. Holstein calves were fed 3.8 L of whole milk, and Jersey calves were fed 2.8 L of whole milk through weaning at 42 d. Calves continued on trial through 63 d. Six treatments were arranged as a 2 × 3 factorial. Calves received either a corn-soybean meal-based starter (21% crude protein and 6% acid detergent fiber; −CSH) or a blend of 85% corn-soybean meal-based starter and 15% CSH (18% crude protein and 14% acid detergent fiber; +CSH) ad libitum. In addition, calves received whole milk with either no supplement (NONE) or supplemented with 3 g/d of mannanoligosaccharide product (MOS) or 4 g/d of live yeast product (YST) through weaning at 42 d. Twelve Holstein steers [n = 6 (per starter type); n = 4 (per supplement type)] were euthanized for collection and examination of rumen tissue samples. Dry matter intake (DMI) was greater for Holstein calves fed +CSH (0.90 kg/d) than −CSH (0.76 kg/d). Final body weight at 63 d of Holstein calves fed +CSH (75.8 kg) was greater than that of those fed −CSH (71.0 kg). Average daily gain (ADG) was greater for Holstein calves fed +CSH (0.58 kg/d) than −CSH (0.52 kg/d). However, Holstein calves fed −CSH had a greater feed efficiency (FE; 0.71 kg of ADG/kg of DMI) than those fed +CSH (0.65 kg of ADG/kg of DMI). Also, Holstein calves fed +CSH had narrower rumen papillae (0.32 mm) compared with those fed −CSH (0.41 mm). There were no significant effects of CSH on DMI, ADG, or FE in Jersey calves. There were no significant effects of YST or MOS on DMI, ADG, FE, or rumen papillae measures in Holstein calves. Jersey calves fed YST or MOS had greater final body weight at 63 d (51.2 kg and 51.0 kg, respectively) than calves fed NONE (47.5 kg). However, there were no significant effects of YST or MOS on DMI, ADG, or FE in Jersey calves.}, number={2}, journal={JOURNAL OF DAIRY SCIENCE}, author={Hill, S. R. and Hopkins, B. A. and Davidson, S. and Bolt, S. M. and Diaz, D. E. and Brownie, C. and Brown, T. and Huntington, G. B. and Whitlow, L. W.}, year={2009}, month={Feb}, pages={790–798} }
 @article{koeberl_pinto_sun_li_kozink_benjamin_demaster_kruse_vaughn_hillman_et al._2008, title={AAV vector-mediated reversal of hypoglycemia in canine and murine glycogen storage disease type Ia}, volume={16}, ISSN={["1525-0016"]}, DOI={10.1038/mt.2008.15}, abstractNote={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.}, number={4}, journal={MOLECULAR THERAPY}, author={Koeberl, Dwight D. and Pinto, Carlos and Sun, Baodong and Li, Songtao and Kozink, Daniel M. and Benjamin, Daniel K., Jr. and Demaster, Amanda K. and Kruse, Meghan A. and Vaughn, Valerie and Hillman, Steven and et al.}, year={2008}, month={Apr}, pages={665–672} }
 @article{sun_young_li_di_brown_salva_li_bird_yan_auten_et al._2008, title={Correction of multiple striated muscles in murine Pompe diseasethrough adeno-associated virus-mediated gene therapy}, volume={16}, ISSN={["1525-0016"]}, DOI={10.1038/mt.2008.133}, abstractNote={Glycogen storage disease type II (Pompe disease; MIM 232300) stems from the deficiency of acid α-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 ∼50% in the heart and quadriceps in GAA-knockout (GAA-KO) mice; furthermore, an AAV2/8 vector containing the hybrid α-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. Glycogen storage disease type II (Pompe disease; MIM 232300) stems from the deficiency of acid α-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 ∼50% in the heart and quadriceps in GAA-knockout (GAA-KO) mice; furthermore, an AAV2/8 vector containing the hybrid α-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. Glycogen storage disease type II (Pompe disease; MIM 232300) is a classical lysosomal storage disease that causes death in infancy as a result of cardiomyopathy and cardiorespiratory failure. The single gene deficiency in acid α-glucosidase (GAA; acid maltase; EC 3.2.1.20) results in lysosomal accumulation of glycogen in various tissues, primarily in heart and skeletal muscle. Pompe disease could be effectively treated by the correction of GAA deficiency in striated muscle. GAA expression with pseudotyped adeno-associated virus (AAV) vectors1Sun B Zhang H Franco LM Young SP Schneider A Bird A et al.Efficacy of an adeno-associated virus 8-pseudotyped vector in glycogen storage disease type II.Mol Ther. 2005; 11: 57-65Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar,2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar,4Mah C Cresawn KO Fraites TJ Jr Pacak CA Lewis MA Zolotukhin I et al.Sustained correction of glycogen storage disease type II using adeno-associated virus serotype 1 vectors.Gene Ther. 2005; 12: 1405-1409Crossref PubMed Scopus (60) Google Scholar,5Cresawn KO Fraites TJ Wasserfall C Atkinson M Lewis M Porvasnik S et al.Impact of humoral immune response on distribution and efficacy of recombinant adeno-associated virus-derived acid alpha-glucosidase in a model of glycogen storage disease type II.Hum Gene Ther. 2005; 16: 68-80Crossref PubMed Scopus (56) Google Scholar or with a helper-dependent adenovirus vector has achieved prolonged efficacy6Kiang A Hartman ZC Liao SX Xu F Serra D Palmer DJ et al.Fully deleted adenovirus persistently expressing GAA accomplishes long-term skeletal muscle glycogen correction in tolerant and nontolerant GSD-II mice.Mol Ther. 2006; 13: 127-134Abstract Full Text Full Text PDF PubMed Scopus (37) Google Scholar in Pompe disease mice. None of the aforementioned vectors completely corrected the glycogen content of all skeletal muscles, and none of these studies evaluated the distal hindlimb muscles. The clinical presentation of Pompe disease resembles that of the muscular dystrophies, featuring weakness of the proximal leg muscles that generalizes to all skeletal muscles.7Hirschhorn R Reuser AJJ Scriver CR Beaudet AL Sly WS Valle D Glycogen storage disease type II: acid α-glucosidase (acid maltase) deficiency.The Metabolic and Molecular Basis for Inherited Disease. McGraw-Hill, New York2001: 3389-3420Google Scholar Gene therapy in the muscular dystrophies represents a unique challenge, because humoral and cytotoxic immune responses occur frequently in response to introduced proteins.8Ferrer A Wells KE Wells DJ Immune responses to dystrophin: implications for gene therapy of Duchenne muscular dystrophy.Gene Ther. 2000; 7: 1439-1446Crossref PubMed Scopus (84) Google Scholar,9Cordier L Gao GP Hack AA McNally EM Wilson JM Chirmule N et al.Muscle-specific promoters may be necessary for adeno-associated virus-mediated gene transfer in the treatment of muscular dystrophies.Hum Gene Ther. 2001; 12: 205-215Crossref PubMed Scopus (129) Google Scholar Cytotoxic T lymphocyte responses against gene therapy vectors have been reduced by substituting muscle-specific regulatory cassettes for ubiquitously active viral promoter/enhancers. An AAV2 vector containing a muscle-specific creatine kinase (MCK) regulatory cassette evoked an attenuated immune response in mdx mice, in comparison with an analogous AAV vector containing the cytomegalovirus (CMV) promoter.10Yuasa K Sakamoto M Miyagoe-Suzuki Y Tanouchi A Yamamoto H Li J et al.Adeno-associated virus vector-mediated gene transfer into dystrophin-deficient skeletal muscles evokes enhanced immune response against the transgene product.Gene Ther. 2002; 9: 1576-1588Crossref PubMed Scopus (113) Google Scholar An AAV2/6 vector containing the MCK CK6 cassette11Hauser MA Robinson A Hartigan-O'Connor D Williams-Gregory D Buskin JN Apone S et al.Analysis of muscle creatine kinase regulatory elements in recombinant adenoviral vectors.Mol Ther. 2000; 2: 16-25Abstract Full Text Full Text PDF PubMed Scopus (80) Google Scholar transduced skeletal muscle with lower efficiency, in comparison with an analogous vector containing a CMV promoter/enhancer; however, the MCK-driven β-galactosidase expression persisted longer than CMV-driven expression.12Gregorevic P Blankinship MJ Allen JM Crawford RW Meuse L Miller DG et al.Systemic delivery of genes to striated muscles using adeno-associated viral vectors.Nat Med. 2004; 10: 828-834Crossref PubMed Scopus (544) Google Scholar An AAV2/6 vector containing another MCK regulatory cassette [CK1 (refs. 13Shield MA Haugen HS Clegg CH Hauschka SD E-box sites and a proximal regulatory region of the muscle creatine kinase gene differentially regulate expression in diverse skeletal muscles and cardiac muscle of transgenic mice.Mol Cell Biol. 1996; 16: 5058-5068Crossref PubMed Scopus (99) Google Scholar,14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar)] produced high-level human GAA (hGAA) expression and glycogen clearance in the injected gastrocnemius muscle, and the analogous AAV2/7 vector partially cleared glycogen storage in multiple muscle groups of GAA-knockout (GAA-KO) mice following intravenous administration.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar AAV2/8 vectors have efficiently transduced striated muscle following systemic delivery in mice.15Nakai H Fuess S Storm TA Muramatsu S Nara Y Kay MA Unrestricted hepatocyte transduction with adeno-associated virus serotype 8 vectors in mice.J Virol. 2005; 79: 214-224Crossref PubMed Scopus (255) Google Scholar As few as 3 × 1011 vector particles transduced the majority of cardiomyocytes.15Nakai H Fuess S Storm TA Muramatsu S Nara Y Kay MA Unrestricted hepatocyte transduction with adeno-associated virus serotype 8 vectors in mice.J Virol. 2005; 79: 214-224Crossref PubMed Scopus (255) Google Scholar,16Wang Z Zhu T Qiao CP Zhou LQ Wang B Zhang J et al.Adeno-associated virus serotype 8 efficiently delivers genes to muscle and heart.Nat Biotechnol. 2005; 23: 321-328Crossref PubMed Scopus (503) Google Scholar More relevant to Pompe disease, an AAV2/1 vector encoding GAA reduced glycogen storage following intravenous administration to neonatal GAA-KO mice.4Mah C Cresawn KO Fraites TJ Jr Pacak CA Lewis MA Zolotukhin I et al.Sustained correction of glycogen storage disease type II using adeno-associated virus serotype 1 vectors.Gene Ther. 2005; 12: 1405-1409Crossref PubMed Scopus (60) Google Scholar Thus, current data from Pompe and muscular dystrophy mice endorse the further investigation of AAV pseudotypes with enhanced muscle tropism in these models.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,4Mah C Cresawn KO Fraites TJ Jr Pacak CA Lewis MA Zolotukhin I et al.Sustained correction of glycogen storage disease type II using adeno-associated virus serotype 1 vectors.Gene Ther. 2005; 12: 1405-1409Crossref PubMed Scopus (60) Google Scholar,17Rutledge EA Halbert CL Russell DW Infectious clones and vectors derived from adeno-associated virus (AAV) serotypes other than AAV type 2.J Virol. 1998; 72: 309-319Crossref PubMed Google Scholar,18Bostick B Ghosh A Yue Y Long C Duan D Systemic AAV-9 transduction in mice is influenced by animal age but not by the route of administration.Gene Ther. 2007; 14: 1605-1609Crossref PubMed Scopus (137) Google Scholar Salva et al. designed a series of highly active muscle-specific regulatory cassettes that were evaluated in striated muscle following systemic delivery via tail vein injections of AAV6 vectors.14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar The most active cassette in a variety of anatomical muscles combined a 190-bp enhancer from the murine α-myosin heavy chain gene with a 570 bp abbreviated MCK regulatory cassette, termed the MHCK7. The latter expressed human placental alkaline phosphatase at very high levels in murine heart and skeletal muscle, exceeding levels achieved with the CMV promoter/enhancer in the heart, and expressed high levels of microdystrophin in skeletal and cardiac muscles of mdx mice.14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar We hypothesized that systemic administration of an AAV vector containing a muscle-specific regulatory cassette could achieve long-term correction of multiple muscles in GAA-KO mice. An AAV vector containing the MHCK7 cassette was pseudotyped with AAV serotypes 7, 8, and 9, and the efficacy of each serotype was evaluated in GAA-KO mice. The transduction of striated muscle with AAV2/8 vectors containing either an abbreviated MCK regulatory cassette (AAV2/8-CK1hGAApA2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar) or a hybrid α-myosin heavy chain enhancer/MCK cassette (AAV2/8-MHCK7hGAApA) were evaluated, following intravenous administration in adult GAA-KO mice (1 × 1011 or 1 × 1012 vector particles/mouse). The former vector contains the CK1 promoter13Shield MA Haugen HS Clegg CH Hauschka SD E-box sites and a proximal regulatory region of the muscle creatine kinase gene differentially regulate expression in diverse skeletal muscles and cardiac muscle of transgenic mice.Mol Cell Biol. 1996; 16: 5058-5068Crossref PubMed Scopus (99) Google Scholar and the latter contains MHCK7.14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar GAA activity and glycogen content in the heart, liver, and skeletal muscles were analyzed 18 weeks following vector administration (Figure 1). GAA activity and glycogen content were significantly affected by both dosage and vector type for all tissues examined (P < 0.05 with a two-way ANOVA). Bonferroni post-test comparisons of the two vectors showed that GAA levels were significantly greater in all tissues treated with the higher dose of AAV2/8-MHCK7hGAApA compared with the equivalent dose of AAV2/8-CK1hGAApA(Figure 1). At the low dose the difference between the two vectors was not significant for any of the tissues, although the mean GAA levels were greater in each tissue for the AAV2/8-MHCK7hGAApA group. The lower GAA activity in the liver following AAV2/8-CK1hGAApA administration suggested a more stringent muscle-restricted expression with the CK1 cassette relative to the MHCK7 cassette (Figure 1a).14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar Real-time reverse transcriptase–PCR revealed that liver expression of GAA was significantly increased with AAV2/8-MHCK7hGAApA, in comparison with AAV2/8-CK1hGAApA, when each was administered at the higher dose (2.2 ± 1.2% of β-actin RNA versus 0.20 ± 0.05%, respectively; P = 0.006). However, secretion of hGAA from transduced muscles and uptake by the liver also could have contributed to the higher GAA activity in the liver following AAV2/8-MHCK7hGAApA administration. Glycogen was significantly lower in all muscles treated with AAV2/8-MHCK7hGAApA at both low and high doses, in comparison with the equivalent dose of AAV2/8-CK1hGAApA (Figure 1b). The presence of residual glycogen accumulation in hindlimb muscles, particularly the gastrocnemius, following AAV2/8 vector administration prompted a comparison with the AAV2/7 and AAV2/9 pseudotypes of AAV-MCHK7hGAApA. The GAA activities of the heart and skeletal muscles were significantly increased following AAV2/8-MHCK7hGAApA or AAV2/9- MHCK7hGAApA administration (P < 0.05), in comparison with AAV2/7-MHCK7hGAApA (Figure 1c). The glycogen contents of the heart and skeletal muscles were significantly decreased following AAV2/8-MHCK7hGAApA or AAV2/9-MHCK7hGAApA administration (P < 0.05), in comparison with AAV2/7-MHCK7hGAApA (Figure 1d). Vector genomes were quantified to allow a comparison of the transduction efficiency with each pseudotype of AAV-MHCK7hGAApA (Figure 1e). Long-term hGAA expression in heart and skeletal muscles was associated with the persistence of up to ∼1 vector genome per nuclear genome in the heart, gastrocnemius, and quadriceps, whereas more than tenfold higher amounts of vector DNA were present in the liver (Figure 1e). The enhanced correction with the AAV2/8 and AAV2/9 vectors, in comparison with AAV2/7, correlated with slightly increased numbers of vector genomes in the heart, quadriceps, and gastrocnemius (Figure 1e); moreover, the AAV2/8 and AAV2/9 vectors demonstrated significantly increased normalized GAA expression in striated muscle and heart (Figure 1f) (P < 0.05). Taken together, these data indicated that the AAV2/8 and AAV2/9 vectors transduced striated muscle more efficiently and produced higher GAA activity in transduced myofibers, in comparison with the AAV2/7 vector. Western blot analysis revealed the ∼76 kd processed form of GAA in the heart, quadriceps, gastrocnemius, and liver of GAA-KO mice following administration of the higher number of particles of AAV2/8-CK1hGAApA and AAV2/8-MHCK6hGAApA, whereas a very low signal was detected in the diaphragm following transduction with AAV2/8-CK1hGAApA (Figure 2, liver GAA not shown). GAA activity and glycogen content were not significantly changed in diaphragm following administration of AAV2/8-CK1hGAApA (Figure 1), consistent with the low activity for the CK1 cassette in the diaphragm.12Gregorevic P Blankinship MJ Allen JM Crawford RW Meuse L Miller DG et al.Systemic delivery of genes to striated muscles using adeno-associated viral vectors.Nat Med. 2004; 10: 828-834Crossref PubMed Scopus (544) Google Scholar,13Shield MA Haugen HS Clegg CH Hauschka SD E-box sites and a proximal regulatory region of the muscle creatine kinase gene differentially regulate expression in diverse skeletal muscles and cardiac muscle of transgenic mice.Mol Cell Biol. 1996; 16: 5058-5068Crossref PubMed Scopus (99) Google Scholar,14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar Glycogen staining revealed the basis for incomplete correction of skeletal gastrocnemius with AAV2/8-MHCK7hGAApA (Figure 1b). Multiple glycogen laden myofibers were detected in the gastrocnemius, consistent with the lack of correction of GAA deficiency within individual myofibers (Figure 3a). The quadriceps contained a higher number of normal-appearing myofibers (Figure 3a, arrows), consistent with the lower glycogen content in the quadriceps (Figure 1b). Thus, the relatively higher residual glycogen in the gastrocnemius could be attributed to the greater prevalence of untransduced myofibers. Lymphocytic infiltrates were absent in skeletal muscle, indicating a lack of cytotoxic T-cell responses in transduced muscle (data not shown). To evaluate the distribution of functionally transduced myofibers, mice were injected with 1 × 1012 vector particles of an AAV2/8 vector carrying a human placental alkaline phosphatase reporter cDNA driven by the MHCK7 regulatory cassette. The highest level of functional transduction was detected in the heart, followed by the quadriceps, whereas fewer transduced myofibers were present in the gastrocnemius (Figure 3b). The soleus and extensor digitalis longus (EDL) were transduced at even lower frequency than the gastrocnemius (less than three positive myofibers/field; data not shown). The low transduction of the soleus and EDL demonstrated a clear limitation for the AAV2/8 pseudotype. The GAA activities of the EDL and soleus were significantly increased following AAV2/9-MHCK7hGAApA administration (P < 0.05), in comparison with AAV2/7-MHCK7hGAApA (Figure 4a). As expected, the glycogen contents of the EDL and soleus were significantly decreased following AAV2/9- MHCK7hGAApA administration (Figure 4b). Thus, the transduction of the small hindlimb muscles was superior for the AAV9-pseudotyped vector. Antibodies against hGAA have prevented efficient cross-correction of GAA deficiency through receptor-mediated uptake, despite the presence of secreted hGAA in the blood.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar,19Ding EY Hodges BL Hu H McVie-Wylie AJ Serra D Migone FK et al.Long-term efficacy after [E1-,polymerase-] adenovirus-mediated transfer of human acid-alpha-glucosidase gene into glycogen storage disease type II knockout mice.Hum Gene Ther. 2001; 12: 955-965Crossref PubMed Scopus (63) Google Scholar Anti-hGAA antibodies were significantly elevated by 6 weeks following administration of both low and high doses of AAV2/8 vector (P < 0.05), regardless of the regulatory cassette type (Figure 5a). Despite the presence of anti-GAA antibodies, Rotarod times were significantly increased (P < 0.05) at 12 and 18 weeks following administration of AAV-MHCK7hGAApA-injected mice for the AAV2/7, AAV2/8, and AAV2/9 pseudotypes (Figure 5b). However, Rotarod times were not significantly increased at 12 or 18 weeks for AAV-CK1hGAApA-treated mice. A glucotetrasaccharide biomarker for increased glycogen storage, Glcα1-6Glcα1-4Glcα1-4Glc, (Glc4), was significantly reduced (P < 0.05) at 18 weeks for the high-dose group of all vector-injected GAA-KO mice, regardless of the regulatory cassette or pseudotype in comparison with phosphate-buffered saline–injected mice (Figure 5c). There was no difference in mean Glc4 levels between the different vector-treated groups. Hence, a significant increase in Rotarod time and reduction of Glc4 biomarker were achieved, despite the presence of an antibody response against hGAA. Wide-spread clearance of glycogen in individual myofibers was demonstrated following AAV2/7 and AAV2/9 vector administration (Figure 6). Fiber typing confirmed the high prevalence of type I, slow-twitch fibers in the soleus of GAA-KO mice, whereas the EDL was comprised mainly of type IIb myofibers (data not shown). The clearance of glycogen vacuolation within multiple myofibers in the EDL and soleus indicated that both type I and IIb myofibers were transduced by the AAV2/9 vector (Figure 6). Gene therapy in the muscular dystrophies will likely require therapeutic gene expression in striated muscle generally, particularly for lethal muscular dystrophies such as Duchenne muscular dystrophy. Pompe disease uniquely responds to infused or secreted therapeutic protein, because unlike other muscular dystrophies Pompe disease is a lysosomal storage disorder amenable to enzyme replacement therapy. Owing to the high enzyme level requirements in enzyme replacement therapy and complicating antibody responses to GAA, muscle-targeted gene therapy is under development in Pompe disease.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,20Raben N Plotz P Byrne BJ Acid alpha-glucosidase deficiency (glycogenosis type II, Pompe disease).Curr Mol Med. 2002; 2: 145-166Crossref PubMed Scopus (192) Google Scholar,21Fraites TJ Jr Schleissing MR Shanely RA Walter GA Cloutier DA Zolotukhin I et al.Correction of the enzymatic and functional deficits in a model of Pompe disease using adeno-associated virus vectors.Mol Ther. 2002; 5: 571-578Abstract Full Text Full Text PDF PubMed Scopus (106) Google Scholar Systemic delivery of an AAV2/8 vector encoding muscle-restricted hGAA achieved significant efficacy; however, the transduction of myofibers was partial, preventing the complete correction of glycogen storage in the gastrocnemius. The analogous AAV2/9 vector demonstrated significantly increased transduction of the small hindlimb muscles, in direct comparison with the AAV2/7 vector. Transduction of the EDL and soleus were inefficient with an AAV2/8 vector encoding human placental alkaline phosphatase, implying that the AAV2/9 pseudotype might have significant advantages with regard to transduction of the distal hindlimb muscles. Type I fibers were more easily cleared of glycogen during enzyme replacement therapy, indicating the need to focus on the transduction of type II fibers in Pompe disease.22Raben N Danon M Gilbert AL Dwivedi S Collins B Thurberg BL et al.Enzyme replacement therapy in the mouse model of Pompe disease.Mol Genet Metab. 2003; 80: 159-169Abstract Full Text Full Text PDF PubMed Scopus (178) Google Scholar,23Fukuda T Ewan L Bauer M Mattaliano RJ Zaal K Ralston E et al.Dysfunction of endocytic and autophagic pathways in a lysosomal storage disease.Ann Neurol. 2006; 59: 700-708Crossref PubMed Scopus (248) Google Scholar Type I myofibers were transduced less efficiently with an AAV2 vector than with an AAV2/6 vector, reflecting the potential advantage of newer serotypes over AAV2.24Blankinship MJ Gregorevic P Allen JM Harper SQ Harper H Halbert CL et al.Efficient transduction of skeletal muscle using vectors based on adeno-associated virus serotype 6.Mol Ther. 2004; 10: 671-678Abstract Full Text Full Text PDF PubMed Scopus (197) Google Scholar,25Pruchnic R Cao B Peterson ZQ Xiao X Li J Samulski RJ et al.The use of adeno-associated virus to circumvent the maturation-dependent viral transduction of muscle fibers.Hum Gene Ther. 2000; 11: 521-536Crossref PubMed Scopus (74) Google Scholar Currently, the AAV2/9 serotype efficiently transduced both type I and IIb myofibers, as well as cardiomyocytes, following intravenous administration. This study further endorses the role of AAV2/9 vectors for gene therapy in Pompe disease, demonstrating significant correction of not only the heart but all skeletal muscles examined. Previously AAV2/9 vectors have transduced striated muscle more efficiently than either an AAV2/1 vector in neonatal mice26Pacak CA Mah CS Thattaliyath BD Conlon TJ Lewis MA Cloutier DE et al.Recombinant adeno-associated virus serotype 9 leads to preferential cardiac transduction in vivo.Circ Res. 2006; 99: e3-e9Crossref PubMed Scopus (316) Google Scholar or an AAV2/8 vector in adult mice.27Inagaki K Fuess S Storm TA Gibson GA McTiernan CF Kay MA et al.Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8.Mol Ther. 2006; 14: 45-53Abstract Full Text Full Text PDF PubMed Scopus (467) Google Scholar Neither of these earlier studies demonstrated significant correction of multiple skeletal muscles in adult mice.26Pacak CA Mah CS Thattaliyath BD Conlon TJ Lewis MA Cloutier DE et al.Recombinant adeno-associated virus serotype 9 leads to preferential cardiac transduction in vivo.Circ Res. 2006; 99: e3-e9Crossref PubMed Scopus (316) Google Scholar,27Inagaki K Fuess S Storm TA Gibson GA McTiernan CF Kay MA et al.Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8.Mol Ther. 2006; 14: 45-53Abstract Full Text Full Text PDF PubMed Scopus (467) Google Scholar The improved efficacy of this new AAV2/9 vector further strengthens preclinical data in favor of clinical trials of gene therapy in Pompe disease. The successful development of gene therapy in GAA-KO mice indicates that curative therapy for Pompe disease may become available in the foreseeable future; however, the efficacy of gene therapy in these experiments was inversely related to the presence of immune responses. Immunocompetent GAA-KO mice produced high titer anti-hGAA immunoglobulin G in response to an AAV vector containing the ubiquitously active CMV promoter/chicken β-actin (CB) promoter, packaged as either AAV2/6 or AAV2/8.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar The CB-containing vector failed to secrete detectable hGAA in the plasma for >2 weeks or to reduce glycogen storage in the muscle of GAA-KO mice.3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar Immune responses to ubiquitously expressed hGAA included lymphocytic infiltrates and activation of CD4+ and CD8+ lymphocytes in injected skeletal muscle, and in the liver following intravenous injection.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar Substitution of the CK1 regulatory cassette in place of the CB promoter prevented CD8+ lymphocytic responses in the injected muscle. Therefore, both antibody production and cytotoxic T lymphocytes were elicited in response to hGAA production from a ubiquitously active CB promoter; however, vectors containing muscle-specific cassettes did not provoke lymphocytic infiltrates in transduced muscles and expressed hGAA for >18 weeks.2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar The latter experiment also demonstrated the imperviousness of muscle-specific hGAA expression to circulating anti-GAA antibodies. The presence of anti-GAA antibodies has prevented cross-correction of untransduced muscle cells,1Sun B Zhang H Franco LM Young SP Schneider A Bird A et al.Efficacy of an adeno-associated virus 8-pseudotyped vector in glycogen storage disease type II.Mol Ther. 2005; 11: 57-65Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar,2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar,3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar,19Ding EY Hodges BL Hu H McVie-Wylie AJ Serra D Migone FK et al.Long-term efficacy after [E1-,polymerase-] adenovirus-mediated transfer of human acid-alpha-glucosidase gene into glycogen storage disease type II knockout mice.Hum Gene Ther. 2001; 12: 955-965Crossref PubMed Scopus (63) Google Scholar implicating the transduction of individual muscle cells as the source of glycogen clearance in the current study. The MHCK7 regulatory cassette has driven highly efficacious hGAA expression in the Pompe disease mouse model in this study. A comparison of AAV2/6 vectors containing either the MHCK7 or the CK1 cassette demonstrated high transgene expression within cardiac muscle and skeletal muscles,14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar which has now been replicated in Pompe disease mice for AAV2/7, AAV2/8, and AAV2/9 vectors. Virtually all myofibers were transduced in the muscles examined with the aforementioned AAV2/6 vector, although transgene expression was reduced in the diaphragm; moreover, transduction was enhanced by the co-administration of the vascular permeabilizing agent vascular endothelial growth factor at lower vector doses.12Gregorevic P Blankinship MJ Allen JM Crawford RW Meuse L Miller DG et al.Systemic delivery of genes to striated muscles using adeno-associated viral vectors.Nat Med. 2004; 10: 828-834Crossref PubMed Scopus (544) Google Scholar The current AAV2/8 vector achieved partial clearance of glycogen from the diaphragm in Pompe mice through the administration of reasonably low particle numbers, increasing the likelihood of translation to clinical applications in Pompe disease. The activity of the MCHK7 regulatory cassette was higher than that for the CK1 cassette in heart, quadriceps, gastrocnemius, and diaphragm, all critical targets for gene therapy in Pompe disease and other forms of muscular dystrophy. The combination of highly active muscle-specific regulatory cassettes and novel AAV serotypes promises to advance gene therapy for muscular dystrophy by providing curative therapy for these devastating disorders. Preparation of pseudotyped AAV vectors. AAV-MHCK7hGAApA contains the MHCK7 regulatory cassette,14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar the hGAA cDNA, and a human growth hormone polyadenylation sequence. The vector plasmid, pAAV-MHCK7hGAApA, was derived from pAAV-CBhGAApA.28Sun B Chen YT Bird A Xu F Hou YX Amalfitano A et al.Packaging of an AAV vector encoding human acid alpha-glucosidase for gene therapy in glycogen storage disease type II with a modified hybrid adenovirus-AAV vector.Mol Ther. 2003; 7: 467-477Abstract Full Text Full Text PDF PubMed Scopus (34) Google Scholar The pAAV-MHCK7hGAApA was digested with KpnI, blunt-ended with the Klenow fragment of DNA polymerase, then digested with XbaI; subsequently, the 5.7-kb blunt/XbaI fragment from pAAV-MHCK7hGAApA was ligated with a 0.8-kb XbaI/blunt SalI fragment containing MHCK7 cassette from αMHCKChAP14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar (provided by Dr. Stephen Hauschka, University of Washington, Seattle, WA). AAV-CK1hGAApA has been described (formerly AAV-MCKhGAApA).2Sun B Zhang H Franco LM Brown T Bird A Schneider A et al.Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter.Mol Ther. 2005; 11: 889-898Abstract Full Text Full Text PDF PubMed Scopus (85) Google Scholar Briefly, 293 cells were transfected with an AAV vector, the AAV packaging plasmid29Gao GP Alvira MR Wang L Calcedo R Johnston J Wilson JM Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy.Proc Natl Acad Sci USA. 2002; 99: 11854-11859Crossref PubMed Scopus (1237) Google Scholar (courtesy of Dr. James M. Wilson, University of Pennsylvania, Philadelphia, PA), and pAdHelper (Stratagene, La Jolla, CA). Cell lysate was harvested 48 hours following infection and freeze-thawed three times, and isolated by sucrose cushion pelleting followed by two cesium chloride gradient centrifugation steps. AAV stocks were dialyzed against three changes of Hanks buffer, and aliquots were stored at −80 °C. The number of vector DNA–containing particles in viral stocks was determined by DNase I digestion, DNA extraction, and Southern blot analysis. The Southern blot signal for vector genomes was quantified by comparison with the signals from standards consisting of AhdI-digested vector plasmid. All viral vector stocks were handled according to Biohazard Safety Level 2 guidelines published by the National Institutes of Health. In vivo analysis of AAV vector. The AAV vector stocks were administered intravenously (via the retroorbital sinus) in 3-month-old GAA-KO mice.30Raben N Nagaraju K Lee E Kessler P Byrne B Lee L et al.Targeted disruption of the acid alpha-glucosidase gene in mice causes an illness with critical features of both infantile and adult human glycogen storage disease type II.J Biol Chem. 1998; 273: 19086-19092Crossref PubMed Scopus (223) Google Scholar At the indicated time points after injection, plasma or tissue samples were obtained and processed as described below. All animal procedures were performed in accordance with Duke University Institutional Animal Care and Use Committee–approved guidelines. Rotarod testing was performed as previously described.1Sun B Zhang H Franco LM Young SP Schneider A Bird A et al.Efficacy of an adeno-associated virus 8-pseudotyped vector in glycogen storage disease type II.Mol Ther. 2005; 11: 57-65Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar GAA activity and glycogen content were analyzed as described, and real-time RT-PCR was performed with standard methods using primers previously described.31Amalfitano A McVie-Wylie AJ Hu H Dawson TL Raben N Plotz P et al.Systemic correction of the muscle disorder glycogen storage disease type II after hepatic targeting of a modified adenovirus vector encoding human acid-alpha-glucosidase.Proc Natl Acad Sci USA. 1999; 96: 8861-8866Crossref PubMed Scopus (133) Google Scholar Western blotting of hGAA was performed as previously described1Sun B Zhang H Franco LM Young SP Schneider A Bird A et al.Efficacy of an adeno-associated virus 8-pseudotyped vector in glycogen storage disease type II.Mol Ther. 2005; 11: 57-65Abstract Full Text Full Text PDF PubMed Scopus (121) Google Scholar using the hGAA monoclonal antibody (courtesy of Genzyme, Framingham, MA). Alkaline phosphatase staining was performed as previously described.14Salva MZ Himeda CL Tai PW Nishiuchi E Gregorevic P Allen JM et al.Design of tissue-specific regulatory cassettes for high-level rAAV-mediated expression in skeletal and cardiac muscle.Mol Ther. 2007; 15: 320-329Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar The enzyme-linked immunosorbent assay was performed as previously described.3Franco LM Sun B Yang X Bird A Zhang H Schneider A et al.Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II.Mol Ther. 2005; 12: 876-884Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar All samples yielded absorbance values that were within the linear range of the assay at this dilution. Urinary Glc4 concentrations were determined relative to creatinine by stable isotope-dilution electrospray tandem mass spectrometry as previously described.32Young SP Stevens RD An Y Chen YT Millington DS Analysis of a glucose tetrasaccharide elevated in Pompe disease by stable isotope dilution-electrospray ionization tandem mass spectrometry.Anal Biochem. 2003; 316: 175-180Crossref PubMed Scopus (59) Google Scholar Statistical analyses. Two-way ANOVAs were performed using the independent variables of vector type and dose of vector, with Bonferroni post-tests for specific comparison between groups (Prism 3.0, GraphPad, San Diego, CA). Comparison of two groups was assessed by a homoscedastic Student's t-test. A P value of <0.05 indicated a significant difference between the observed values for each group. This work was supported by National Institutes of Health grant R01 HL081122-01A1 from the National Heart, Lung, and Blood Institute. D.D.K. was supported by the Muscular Dystrophy Association and Genzyme Corporation. B.S. was supported by a Development Grant from the Muscular Dystrophy Association. Development and evaluation of the MCK regulatory cassettes were supported by National Institutes of Health grants RO1 AR18860, 1 U54 HD047175, and R24 HL64387, and the Muscular Dystrophy Association to S.D.H. GAA-KO mice were provided courtesy of Nina Raben at the National Institutes of Health (Bethesda, MD). The AAV8 packaging plasmid, p5E18-VD 2/8, was provided courtesy of James M. Wilson at the University of Pennsylvania (Philadelphia, PA).}, number={8}, journal={MOLECULAR THERAPY}, author={Sun, Baodong and Young, Sarah P. and Li, Ping and Di, Chunhui and Brown, Talmage and Salva, Maja Z. and Li, Songtao and Bird, Andrew and Yan, Zhen and Auten, Richard and et al.}, year={2008}, month={Aug}, pages={1366–1371} }
 @article{mackillop_olby_linder_brown_2007, title={Intramedullary cavernous malformation of the spinal cord in two dogs}, volume={44}, ISSN={["1544-2217"]}, DOI={10.1354/vp.44-4-528}, abstractNote={Intramedullary cavernous malformations (CVMs) of the spinal cord were diagnosed in 2 adult dogs that presented for paraparesis. An intramedullary spinal cord lesion was identified on a myelogram in the first dog, and expansion of the vertebral canal was evident on radiographs in the second. Extensive intraparenchymal hemorrhage was found on gross postmortem examination in both dogs, and a distinct lobulated intramedullary mass was evident in the second dog. Microscopically, both lesions were composed of dilated, thin-walled vascular channels with little-to-no intervening neural parenchyma. Both dogs had evidence of channel thrombosis along with perilesional hemorrhage and hemosiderin accumulation. The second dog had additional degenerative changes, including thickened fibrous channel walls with hyalinization, foci of mineralization, and occasional tongues of entrapped gliotic neuropil. CVMs appear to be an uncommon cause of both acute and chronic spinal cord disease in the dog.}, number={4}, journal={VETERINARY PATHOLOGY}, author={Mackillop, E. and Olby, N. J. and Linder, K. E. and Brown, T. T.}, year={2007}, month={Jul}, pages={528–532} }
 @article{koeberl_sun_damodaran_brown_millington_benjamin_bird_schneider_hillman_jackson_et al._2006, title={Early, sustained efficacy of adeno-associated virus vector-mediated gene therapy in glycogen storage disease type Ia}, volume={13}, ISSN={["1476-5462"]}, DOI={10.1038/sj.gt.3302774}, abstractNote={The deficiency of glucose-6-phosphatase (G6Pase) underlies life-threatening hypoglycemia and growth retardation in glycogen storage disease type Ia (GSD-Ia). An adeno-associated virus (AAV) vector encoding G6Pase was pseudotyped as AAV8 and administered to 2-week-old GSD-Ia mice (n = 9). Median survival was prolonged to 7 months following vector administration, in contrast to untreated GSD-Ia mice that survived for only 2 weeks. Although GSD-Ia mice were initially growth-retarded, treated mice increased fourfold in weight to normal size. Blood glucose was partially corrected by 2 weeks following treatment, whereas blood cholesterol normalized. Glucose-6-phosphatase activity was partially corrected to 25% of the normal level at 7 months of age in treated mice, and blood glucose during fasting remained lower in treated, affected mice than in normal mice. Glycogen storage was partially corrected in the liver by 2 weeks following treatment, but reaccumulated to pre-treatment levels by 7 months old (m.o.). Vector genome DNA decreased between 3 days and 3 weeks in the liver following vector administration, mainly through the loss of single-stranded genomes; however, double-stranded vector genomes were more stable. Although CD8+ lymphocytic infiltrates were present in the liver, partial biochemical correction was sustained at 7 m.o. The development of efficacious AAV vector-mediated gene therapy could significantly reduce the impact of long-term complications in GSD-Ia, including hypoglycemia, hyperlipidemia and growth failure.}, number={17}, journal={GENE THERAPY}, author={Koeberl, D. D. and Sun, B. D. and Damodaran, T. V. and Brown, T. and Millington, D. S. and Benjamin, D. J., Jr. and Bird, A. and Schneider, A. and Hillman, S. and Jackson, M. and et al.}, year={2006}, month={Sep}, pages={1281–1289} }
 @article{sun_zhang_benjamin_brown_bird_young_mcvie-wylie_chen_koeberl_2006, title={Enhanced efficacy of an AAV vector encoding chimeric, highly secreted acid alpha-glucosidase in glycogen storage disease type II}, volume={14}, ISSN={["1525-0024"]}, DOI={10.1016/j.ymthe.2006.08.001}, abstractNote={<h2>Abstract</h2> Glycogen storage disease type II (GSD-II; Pompe disease; MIM 232300) is an inherited muscular dystrophy caused by deficiency in the activity of the lysosomal enzyme acid α-glucosidase (GAA). We hypothesized that chimeric GAA containing an alternative signal peptide could increase the secretion of GAA from transduced cells and enhance the receptor-mediated uptake of GAA in striated muscle. The relative secretion of chimeric GAA from transfected 293 cells increased up to 26-fold. Receptor-mediated uptake of secreted, chimeric GAA corrected cultured GSD-II patient cells. High-level hGAA was sustained in the plasma of GSD-II mice for 24 weeks following administration of an AAV2/8 vector encoding chimeric GAA; furthermore, GAA activity was increased and glycogen content was significantly reduced in striated muscle and in the brain. Administration of only 1×10<sup>10</sup> vector particles increased GAA activity in the heart and diaphragm for >18 weeks, whereas 3×10<sup>10</sup> vector particles increased GAA activity and reduced glycogen content in the heart, diaphragm, and quadriceps. Furthermore, an AAV2/2 vector encoding chimeric GAA produced secreted hGAA for >12 weeks in the majority of treated GSD-II mice. Thus, chimeric, highly secreted GAA enhanced the efficacy of AAV vector-mediated gene therapy in GSD-II mice.}, number={6}, journal={MOLECULAR THERAPY}, author={Sun, Baodong and Zhang, Haoyue and Benjamin, Daniel K., Jr. and Brown, Talmage and Bird, Andrew and Young, Sarah P. and McVie-Wylie, Alison and Chen, Y. -T. and Koeberl, Dwight D.}, year={2006}, month={Dec}, pages={822–830} }
 @article{sun_zhang_franco_brown_bird_schneider_koeberl_2005, title={Correction of glycogen storage disease type II by an adeno-associated virus vector containing a muscle-specific promoter}, volume={11}, ISSN={["1525-0024"]}, DOI={10.1016/j.ymthe.2005.01.012}, abstractNote={Glycogen storage disease type II (Pompe disease) causes death in infancy from cardiorespiratory failure due to acid alpha-glucosidase (GAA; acid maltase) deficiency. An AAV2 vector pseudotyped as AAV6 (AAV2/6 vector) transiently expressed high-level human GAA in GAA-knockout (GAA-KO) mice without reducing glycogen storage; however, in immunodeficient GAA-KO/SCID mice the AAV2/6 vector expressed high-level GAA and reduced the glycogen content of the injected muscle for 24 weeks. A CD4+/CD8+ lymphocytic infiltrate was observed in response to the AAV2/6 vector in immunocompetent GAA-KO mice. When a muscle-specific creatine kinase promoter was substituted for the CB promoter (AAV-MCKhGAApA), that AAV2/6 vector expressed high-level GAA and reduced glycogen content in immunocompetent GAA-KO mice. Muscle-restricted expression of hGAA provoked only a humoral (not cellular) immune response. Intravenous administration of a high number of particles of AAV-MCKhGAApA as AAV2/7 reduced the glycogen content of the heart and skeletal muscle and corrected individual myofibers in immunocompetent GAA-KO mice 24 weeks postinjection. In summary, persistent correction of muscle glycogen content was achieved with an AAV vector containing a muscle-specific promoter in GAA-KO mice, and this approach should be considered for muscle-targeted gene therapy in Pompe disease.}, number={6}, journal={MOLECULAR THERAPY}, author={Sun, BD and Zhang, HY and Franco, LM and Brown, T and Bird, A and Schneider, A and Koeberl, DD}, year={2005}, month={Jun}, pages={889–898} }
 @article{franco_sun_yang_bird_zhang_schneider_brown_young_clay_amalfitano_et al._2005, title={Evasion of immune responses to introduced human acid alpha-glucosidase by liver-restricted expression in glycogen storage disease type II}, volume={12}, ISSN={["1525-0024"]}, DOI={10.1016/j.ymthe.2005.04.024}, abstractNote={Glycogen storage disease type II (GSD-II; Pompe disease) is caused by a deficiency of acid alpha-glucosidase (GAA; acid maltase) and manifests as muscle weakness, hypertrophic cardiomyopathy, and respiratory failure. Adeno-associated virus vectors containing either a liver-specific promoter (LSP) (AAV-LSPhGAApA) or a hybrid CB promoter (AAV-CBhGAApA) to drive human GAA expression were pseudotyped as AAV8 and administered to immunocompetent GAA-knockout mice. Secreted hGAA was detectable in plasma between 1 day and 12 weeks postadministration with AAV-LSPhGAApA and only from 1 to 8 days postadministration for AAV-CBGAApA. No anti-GAA antibodies were detected in response to AAV-LSPhGAApA (<1:200), whereas AAV-CBhGAApA provoked an escalating antibody response starting 2 weeks postadministration. The LSP drove approximately 60-fold higher GAA expression than the CB promoter in the liver by 12 weeks following vector administration. Furthermore, the detected cellular immunity was provoked by AAV-CBhGAApA, as detected by ELISpot and CD4+/CD8+ lymphocyte immunodetection. GAA activity was increased to higher than normal and glycogen content was reduced to essentially normal levels in the heart and skeletal muscle following administration of AAV-LSPhGAApA. Therefore, liver-restricted GAA expression with an AAV vector evaded immunity and enhanced efficacy in GSD-II mice.}, number={5}, journal={MOLECULAR THERAPY}, author={Franco, LM and Sun, BD and Yang, XY and Bird, A and Zhang, HY and Schneider, A and Brown, T and Young, SP and Clay, TM and Amalfitano, A and et al.}, year={2005}, month={Nov}, pages={876–884} }
 @misc{fuji_patton_steinbach_schulman_bradley_brown_wilson_summers_2005, title={Feline systemic reactive angioendotheliomatosis: Eight cases and literature review}, volume={42}, ISSN={["1544-2217"]}, DOI={10.1354/vp.42-5-608}, abstractNote={A rare, multisystemic intravascular proliferative disorder was identified postmortem in eight cats. The majority of these cats died or were euthanized following episodes of dyspnea, lethargy, and anorexia. Microscopic examination revealed occlusive, intraluminal proliferations of spindle cells within small vessels. The heart was consistently involved, and myocardial dysfunction was the probable cause of illness in all cats. Immunohistochemically, the majority of intravascular cells expressed von Willebrand factor, and a smaller number expressed smooth muscle actin, compatible with a dual population of endothelial cells and pericytes, suggesting a reactive rather than a neoplastic process. Four cases of a similar feline vascular disorder from the veterinary literature are reviewed. The histopathology resembles reactive angioendotheliomatosis in humans, a benign cutaneous intravascular endothelial and pericytic proliferative condition. However, in contrast, this feline disease is multisystemic and fatal. We propose the name “feline systemic reactive angioendotheliomatosis” for this unique, idiopathic disorder of domestic cats.}, number={5}, journal={VETERINARY PATHOLOGY}, author={Fuji, RN and Patton, KM and Steinbach, TJ and Schulman, FY and Bradley, GA and Brown, TT and Wilson, EA and Summers, BA}, year={2005}, month={Sep}, pages={608–617} }
 @article{breitschwerdt_debroy_mexas_brown_remick_2005, title={Isolation of necrotoxigenic Escherichia coli from a dog with hemorrhagic pneumonia}, volume={226}, ISSN={0003-1488}, url={http://dx.doi.org/10.2460/javma.2005.226.2016}, DOI={10.2460/javma.2005.226.2016}, abstractNote={A 7-month-old sexually intact male Cocker Spaniel was admitted to the North Carolina State University Veterinary Teaching Hospital for evaluation of lethargy, panting, and excessive salivation that had become progressively severe during a 5-hour period. Despite intensive medical care, the dog died within the first 24 hours of hospitalization, and death was attributed to acute, severe, necrotizing pneumonia. Lung tissue collected at necropsy by use of swabs was cultured and yielded an isolate of Escherichia coli; because of the rapid progression of illness in an otherwise healthy dog, the isolate underwent virulence typing and was determined to be a necrotoxigenic E. coli. Necrotoxigenic E. coli produce a toxin called cytotoxic necrotizing factor and are known to be involved in extraintestinal infections, including urinary tract infection, in humans and animals. Virulence typing of E. coli isolates from dogs with peracute pneumonia is recommended to further characterize the epidemiologic characteristics and public health importance of necrotoxigenic E. coli.}, number={12}, journal={Journal of the American Veterinary Medical Association}, publisher={American Veterinary Medical Association (AVMA)}, author={Breitschwerdt, Edward B. and DebRoy, Chitrita and Mexas, Angela M. and Brown, Talmage T. and Remick, Amera K.}, year={2005}, month={Jun}, pages={2016–2019} }
 @article{hill_hopkins_davidson_bolt_diaz_brownie_brown_huntington_whitlow_2005, title={Technical note: Technique for dissection and analysis of the rumen in young calves}, volume={88}, ISSN={["1525-3198"]}, DOI={10.3168/jds.S0022-0302(05)72691-6}, abstractNote={This paper discusses a technique used to evaluate rumen development in young calves, including removal, dissection, and analysis of tissue. The method allowed for examination of the different sacs of the rumen (dorsal, ventral, cranial, and caudal) using scanning electron microscopy to measure papillae denseness and histology slides to measure papillae length and width. Computer software was used to produce accurate measurements of papillae. The rumens of young calves were dissected, and samples were taken from the cranial, caudal, ventral, and dorsal sections. Calves were part of a nutrition research study, and dietary treatments did have an effect on development measurements such as length, width, and papillae denseness.}, number={1}, journal={JOURNAL OF DAIRY SCIENCE}, author={Hill, SR and Hopkins, BA and Davidson, S and Bolt, SM and Diaz, DE and Brownie, C and Brown, T and Huntington, GB and Whitlow, LW}, year={2005}, month={Jan}, pages={324–326} }
 @article{olby_blot_thibaud_phillips_dp o'brien_burr_berg_brown_breen_2004, title={Cerebellar cortical degeneration in adult American Staffordshire Terriers}, volume={18}, ISSN={["1939-1676"]}, DOI={10.1892/0891-6640(2004)18<201:CCDIAA>2.0.CO;2}, abstractNote={Adult-onset cerebellar cortical degeneration recently has been reported in American Staffordshire Terriers. We describe the clinical and histopathologic features of this disease and examine its mode of inheritance in 63 affected dogs. The age at which neurologic deficits 1st were recognized varied from 18 months to 9 years, with the majority of dogs presented to veterinarians between 4 and 6 years of age. Time from onset of clinical signs to euthanasia varied from 6 months to 6.5 years, with the majority of affected dogs surviving from 2 to 4 years. Initial neurologic findings included stumbling, truncal sway, and ataxia exacerbated by lifting the head up and negotiating stairs. Signs progressed to obvious ataxia characterized by dysmetria, nystagmus, coarse intention tremor, variable loss of menace reaction, marked truncal sway, and falling with transient opisthotonus. With continued progression, dogs became unable to walk without falling repeatedly. Cerebellar atrophy was visible on magnetic resonance images and on gross pathology. Histopathologic findings included marked loss of Purkinje neurons with thinning of the molecular and granular layers and increased cellularity of the cerebellar nuclei. The closest common ancestor of the dogs was born in the 1950s and inheritance was most consistent with an autosomal recessive mode of transmission with a prevalence estimated at 1 in 400 dogs. This inherited disease is comparable to the group of diseases known as spinocerebellar ataxias in humans. Many spinocerebellar ataxias in humans are caused by nucleotide repeats, and this genetic aberration merits investigation as a potential cause of the disease in American Staffordshire Terriers.}, number={2}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Olby, N and Blot, S and Thibaud, JL and Phillips, J and DP O'Brien and Burr, J and Berg, J and Brown, T and Breen, M}, year={2004}, pages={201–208} }
 @article{noureddine_harder_olby_spaulding_brown_2004, title={Ultrasonographic appearance of Dandy Walker-like Syndrome in a Boston terrier}, volume={45}, DOI={10.1111/j.1740-8260.2004.04064.x}, number={4}, journal={Veterinary Radiology & Ultrasound}, author={Noureddine, C. and Harder, R. and Olby, Natasha and Spaulding, K. and Brown, T.}, year={2004}, pages={336–339} }
 @article{beaty_jackson_peterson_bird_brown_benjamin_juopperi_kishnani_boney_chen_et al._2002, title={Delivery of glucose-6-phosphatase in a canine model for glycogen storage disease, type Ia, with adeno-associated virus (AAV) vectors}, volume={9}, ISSN={["0969-7128"]}, DOI={10.1038/sj.gt.3301728}, number={15}, journal={GENE THERAPY}, author={Beaty, RM and Jackson, M and Peterson, D and Bird, A and Brown, T and Benjamin, DK and Juopperi, T and Kishnani, P and Boney, A and Chen, YT and et al.}, year={2002}, month={Aug}, pages={1015–1022} }
 @article{feng_tompkins_xu_brown_laster_zhang_mccaw_2002, title={Thymocyte and peripheral blood T lymphocyte subpopulation changes in piglets following in utero infection with porcine reproductive and respiratory syndrome virus}, volume={302}, ISSN={["0042-6822"]}, DOI={10.1006/viro.2002.1650}, abstractNote={Piglets infected in utero with porcine reproductive and respiratory syndrome virus (PRRSV) are born severely immunocompromised. In this article we more closely examine the effects of in utero PRRSV infection on circulating and thymic T cell populations. Numbers of CD4+, CD8+, and dual-positive lymphocytes were quantitated in circulation and in the thymus during the 2 weeks following birth. At birth we found that the number of circulating lymphocytes was suppressed by 60%. Lymphocyte numbers were also suppressed by 42% at day 7, but by day 14 the number of lymphocytes had rebounded and was actually 47% greater than controls. At birth and day 7, a drop in the number of CD4+ cells could partially explain the suppression we observed, while the rebound in total lymphocyte numbers seen at day 14 was due to a nearly fourfold increase in the number of circulating CD8+ cells. As a result, the normal CD4+:CD8+ ratio of between 1.4 and 2.2 for neonatal pigs was reduced to 0.1-0.5. The thymuses of infected piglets were found to be 50% smaller than those of control pigs and were characterized by cortical involution and severe cortical depletion of thymocytes. Analysis of the population of thymocytes revealed that double-positive thymocytes were suppressed to a greater degree than either single positive subpopulation. In addition, we show that the number of thymocytes undergoing apoptosis was increased twofold in piglets infected with PRRSV. Taken together, these results help explain the dramatic immunosuppression observed in neonatal animals infected in utero with PRRSV.}, number={2}, journal={VIROLOGY}, author={Feng, WH and Tompkins, MB and Xu, JS and Brown, TT and Laster, SM and Zhang, HX and McCaw, MB}, year={2002}, month={Oct}, pages={363–372} }
 @article{kishnani_faulkner_vancamp_jackson_brown_boney_koeberl_chen_2001, title={Canine model and genomic structural organization of glycogen storage disease type Ia (GSD Ia)}, volume={38}, ISSN={["0300-9858"]}, DOI={10.1354/vp.38-1-83}, abstractNote={A canine model of glycogen storage disease Ia (GSD Ia), similar clinically, biochemically, and pathologically to the human disease, was established by crossbreeding Maltese and Beagle dogs carrying a mutated, defective glucose-6-phosphatase (G-6-Pase) gene. Ten puppies were born in three litters from these crossbreedings. Six were homozygous for the previously described M121I GSD Ia mutation. Of these six affecteds, two were stillborn, and one died at 2, 32, and 60 days of life, respectively (puppies A, B, C, D, E), while one is alive at age 15 months (puppy F). Affected puppies exhibited tremors, weakness, and neurologic signs when hypoglycemic. They had postnatal growth retardation and progressive hepatomegaly. Biochemical abnormalities included fasting hypoglycemia, hyperlactacidemia, hypercholesterolemia, hypertriglyceridemia, and hyperuricemia. Microscopic examination of tissues from affected puppies showed diffuse, marked hepatocellular vacuolation, with distended clear hepatocytes and central to marginally located rounded nuclei. In the kidneys of puppies D and E, there was segmental glomerular sclerosis and vacuolation of proximal convoluted tubular epithelium. Biochemical analysis revealed increased liver glycogen content and isolated markedly reduced G-6-Pase enzyme activity in liver and kidney. The canine G-6-Pase gene was characterized by screening a canine genomic library. It spans approximately 11.8 kb and consists of five exons with >90% amino acid sequence homology to the derived human sequence. The first 1.5 kb of the 5' region was sequenced and contains several putative response element motifs homologous to the human 5' region. Establishment of this canine colony of GSD Ia that closely resembles human disease and isolation of the canine genomic gene provides an excellent model for studying pathophysiology and long-term complications and an opportunity to develop novel therapeutic approaches such as drug and gene therapy.}, number={1}, journal={VETERINARY PATHOLOGY}, author={Kishnani, PS and Faulkner, E and VanCamp, S and Jackson, M and Brown, T and Boney, A and Koeberl, D and Chen, YT}, year={2001}, month={Jan}, pages={83–91} }
 @article{pappalardo_brown_tompkins_breitschwerdt_2001, title={Immunopathology of Bartonella vinsonii (berkhoffii) in experimentally infected dogs}, volume={83}, ISSN={["1873-2534"]}, DOI={10.1016/S0165-2427(01)00372-5}, abstractNote={Following natural infection with Bartonella, dogs and humans develop comparable disease manifestations including endocarditis, peliosis hepatis, and granulomatous disease. As the immunologic response to infection in these hosts has not been clearly established, data presented here was derived from the experimental infection of six specific pathogen free (SPF) beagles with a known pathogenic strain of Bartonella. Six dogs were inoculated intravenously with 10(9)cfu of B. vinsonii ssp. berkhoffii and six control dogs were injected intravenously with an equivalent volume of sterile saline. Despite production of substantial levels of specific antibody, blood culture and molecular analyses indicated that Bartonella established chronic infection in these dogs. Flow cytometric analysis of monocytes indicated impaired bacterial phagocytosis during chronic Bartonella infection. There was also a sustained decrease in the percentage of CD8+ lymphocytes in the peripheral blood. Moreover, modulation of adhesion molecule expression (downregulation of L-selectin, VLA-4, and LFA-1) on CD8+ lymphocytes suggested quantitative and qualitative impairment of this cell subset in Bartonella-infected dogs. When compared with control dogs, flow cytometric analysis of lymph node (LN) cells from B. vinsonii infected dogs revealed an expanded population of CD4+ T cells with an apparent naïve phenotype (CD45RA+/CD62L+/CD49D(dim)). However, fewer B cells from infected dogs expressed cell-surface MHC II, implicating impaired antigen presentation to helper T cells within LN. Taken together, results from this study indicate that B. vinsonii establishes chronic infection in dogs which may result in immune suppression characterized by defects in monocytic phagocytosis, an impaired subset of CD8+ T lymphocytes, and impaired antigen presentation within LN.}, number={3-4}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Pappalardo, BL and Brown, TT and Tompkins, M and Breitschwerdt, EB}, year={2001}, month={Dec}, pages={125–147} }
 @article{feng_laster_tompkins_brown_xu_altier_gomez_benfield_mccaw_2001, title={In utero infection by porcine reproductive and respiratory syndrome virus is sufficient to increase susceptibility of piglets to challenge by Streptococcus suis type II}, volume={75}, ISSN={["1098-5514"]}, DOI={10.1128/JVI.75.10.4889-4895.2001}, abstractNote={Porcine reproductive and respiratory syndrome (PRRS) consistently elevates the frequency of disease and mortality in young pigs. Many different secondary bacterial diseases occur in PRRS virus (PRRSV)-infected pigs. However, to date, establishing a reproducible experimental model of PRRSV infection in weaned pigs, with subsequent clinical disease following secondary bacterial challenge, has been difficult. PRRSV is frequently isolated during outbreaks from weak-born piglets affected by secondary bacterial diseases. This study was performed to investigate the potential role of intrauterine PRRSV infection on piglet susceptibility to secondary bacterial infection. PRRSV-free pregnant sows were intranasally infected at 98 days of gestation with PRRSV strain SD 23983. All piglets born to the PRRSV-infected sows were viremic. Piglets were removed from the sows at birth and deprived of colostrum. Piglets from PRRSV-infected and noninfected sows were randomly assigned to Streptococcus suis challenge or control subgroups. At 5 days of age, piglets were challenged intranasally with strain MN 87555 of S. suis type II. Total and differential leukocyte counts were performed on blood samples collected at 3 days of age. The numbers of leukocytes, lymphocytes, and monocytes were significantly reduced in the PRRSV-infected piglets. Lesions were observed in bone marrow, brain, lung, heart, spleen, lymph node, tonsil, and thymus of PRRSV-infected piglets. Thymus/body weight ratios of in utero PRRSV-infected piglets were significantly reduced compared to those of non-PRRSV-infected piglets, and thymic lesions were characterized by severe cortical depletion of thymocytes. Lesions were not observed in piglets born to PRRSV-free sows. Overall, 20 out of 22 piglets in the PRRSV-S. suis dual-infection group died within 1 week after challenge with S. suis (10 of 11 in each of two trials). This contrasts with 1 of 18 piglets in the PRRSV-infection-only group and 5 of 23 piglets in the S. suis-challenge-only group (1 of 12 in trial 1 and 4 of 11 in trial 2). No piglets died in the uninfected control groups. Most of the piglets in the PRRSV-S. suis dual-infection group developed suppurative meningitis. S. suis type II was recovered from their brains and joints. These results indicate that in utero infection by PRRSV makes piglets more susceptible to infection and disease following challenge by S. suis type II. In utero infection by PRRSV may provide a useful model to study the interaction between PRRSV and bacterial coinfections in piglets.}, number={10}, journal={JOURNAL OF VIROLOGY}, author={Feng, WH and Laster, SM and Tompkins, M and Brown, T and Xu, JS and Altier, C and Gomez, W and Benfield, D and McCaw, MB}, year={2001}, month={May}, pages={4889–4895} }
 @article{chittick_rotstein_brown_wolfe_2001, title={Pyometra and uterine adenocarcinoma in a melengestrol acetate- implanted captive coati (Nasua nasua)}, volume={32}, DOI={10.1638/1042-7260(2001)032[0245:pauaia]2.0.co;2}, abstractNote={A 9-yr and 3-mo-old captive female coati (Nasua nasua) was implanted with melengestrol acetate for contraception for 4.5 yr prior to presentation. During her annual examination, purulent vaginal discharge and a palpably prominent uterus were identified. Ancillary diagnostic tests including hematology, cystocentesis, radiographs, and abdominal ultrasound were consistent with pyometra. An ovariohysterectomy was performed and histologic examination revealed pyometra and uterine adenocarcinoma, similar to pathology that has been associated with melengestrol acetate contraception in felids, canids, and primates. Given the potential association between melengestrol acetate and uterine pathology in this case, we recommend caution with melengestrol acetate use in procyonids.}, number={2}, journal={Journal of Zoo and Wildlife Medicine}, author={Chittick, E. and Rotstein, D. and Brown, T. and Wolfe, B.}, year={2001}, pages={245–251} }
 @article{pappalardo_brown_gebhardt_sontakke_breitschwerdt_2000, title={Cyclic CD8+lymphopenia in dogs experimentally infected with Bartonella vinsonii subsp berkhoffii}, volume={75}, ISSN={["0165-2427"]}, DOI={10.1016/S0165-2427(00)00182-3}, abstractNote={Until recently, it was presumed that Bartonella vinsonii only infected voles, a species of North American rodents. In April of 1993, however, our laboratory isolated a novel subspecies of B. vinsonii (B. vinsonii subsp. berkhoffii) from the blood of a dog diagnosed with vegetative valvular endocarditis. Subsequently, based on a seroepidemiologic survey of dogs from North Carolina and Virginia presenting for a variety of medical problems, we found evidence supporting a potentially important association between B. vinsonii and Ehrlichia canis co-infection in dogs. In the following study, eight dogs were infected with B. vinsonii: four specific pathogen free dogs and four dogs that had previously been infected with E. canis. Flow cytometric analysis of peripheral blood lymphocytes revealed a cyclic elevation of the CD4/CD8 T-cell ratio that correlated with cyclic CD8+ lymphopenia in all dogs infected with B. vinsonii, regardless of prior exposure to E. canis.}, number={1-2}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Pappalardo, BL and Brown, T and Gebhardt, D and Sontakke, S and Breitschwerdt, EB}, year={2000}, month={Jun}, pages={43–57} }
 @article{luginbuhl_poore_spears_brown_2000, title={Effect of dietary copper level on performance and copper status of growing meat goats}, volume={16}, number={2000}, journal={Sheep & Goat Research Journal}, author={Luginbuhl, J. M. and Poore, M. H. and Spears, J. W. and Brown, T. T.}, year={2000}, pages={65–71} }
 @article{henry_schmader_brown_miller_howell_daley_hamilton_2000, title={Enhanced green fluorescent protein as a marker for localizing murine cytomegalovirus in acute and latent infection}, volume={89}, ISSN={["0166-0934"]}, DOI={10.1016/S0166-0934(00)00202-0}, abstractNote={A recombinant murine cytomegalovirus (mCMV) that expresses enhanced green fluorescent protein (EGFP) under control of the native immediate-early 1/3 promoter was constructed to detect directly sites of viral activity in latent and reactivated infections. The recombinant virus had acute and latent infection characteristics similar to those of wild-type mCMV. Rare green-fluorescing foci were observed in paraffin sections from lungs and spleens infected latently. Positive immunoperoxidase staining for EGFP in sections of the same lung tissues suggests that these cells may be sites of restricted viral gene expression. EGFP was detected easily in tissue explants reactivating from latent infection in vitro. Morphology and adhesion characteristics of fluorescing cells suggest that viral reactivation occurs in tissue macrophages in explant cultures. The observations presented in this study demonstrate the usefulness of EGFP-expressing recombinants as tools for direct tracking of mCMV activity in vivo and in vitro.}, number={1-2}, journal={JOURNAL OF VIROLOGICAL METHODS}, author={Henry, SC and Schmader, K and Brown, TT and Miller, SE and Howell, DN and Daley, GG and Hamilton, JD}, year={2000}, month={Sep}, pages={61–73} }
 @article{pappalardo_brown_gookin_morrill_breitschwerdt_2000, title={Granulomatous disease associated with Bartonella infection in 2 dogs}, volume={14}, ISSN={["0891-6640"]}, DOI={10.1892/0891-6640(2000)014<0037:GDAWII>2.3.CO;2}, abstractNote={Shortly after removal of an engorged tick from the left ear, a 4-year-old Greyhound was referred for evaluation of fever and a rapidly enlarging mass in the region of the left submandibular lymph node. Histopathologic evaluation of the lymph node resulted in a diagnosis of severe granulomatous lymphadenitis. An 11-year-old mixed-breed dog was referred for evaluation of a 6-week history of serous nasal discharge. Histologic examination of a surgical biopsy from a nasal mass indicated multifocal granulomatous inflammation with fibrosis. Serum samples obtained from both dogs were reactive by immunofluorescent assay to Bartonella vinsonii subsp. berkhoffii antigens (reciprocal titers of 128). Although Bartonella organisms were not isolated by lysis centrifugation blood culture, Bartonella DNA was amplified from tissue samples obtained from each dog (lymph node biopsy from dog 1 and nasal biopsy from dog 2) using primers that amplify a portion of the 16S rRNA gene followed by Southern blot hybridization using a genus-specific probe. Additionally, restriction fragment length polymorphism (RFLP) analysis of a Bartonella-specific citrate synthase gene product obtained from dog 2 resulted in a restriction pattern identical to B. vinsonii subsp. berkhoffii. This is the 1st report of granulomatous disease in dogs associated with Bartonella infection.}, number={1}, journal={JOURNAL OF VETERINARY INTERNAL MEDICINE}, author={Pappalardo, BL and Brown, T and Gookin, JL and Morrill, CL and Breitschwerdt, EB}, year={2000}, pages={37–42} }
 @article{breitschwerdt_atkins_brown_kordick_snyder_1999, title={Bartonella vinsonii subsp berkhoffi and related members of the alpha subdivision of the Proteobacteria in dogs with cardiac arrhythmias, endocarditis, or myocarditis}, volume={37}, number={11}, journal={Journal of Clinical Microbiology}, author={Breitschwerdt, E. B. and Atkins, C. E. and Brown, T. T. and Kordick, D. L. and Snyder, P. S.}, year={1999}, pages={3618–3626} }
 @article{kordick_brown_shin_breitschwerdt_1999, title={Clinical and pathologic evaluation of chronic Bartonella henselae or Bartonella clarridgeiae infection in cats}, volume={37}, number={5}, journal={Journal of Clinical Microbiology}, author={Kordick, D. L. and Brown, T. T. and Shin, K. and Breitschwerdt, E. B.}, year={1999}, pages={1536–1547} }
 @article{rogers_poore_ferko_brown_deaton_bawden_1999, title={Dental wear and growth performance in steers fed sweetpotato cannery waste}, volume={214}, number={5}, journal={Journal of the American Veterinary Medical Association}, author={Rogers, G. M. and Poore, M. H. and Ferko, B. L. and Brown, T. T. and Deaton, T. G. and Bawden, J. W.}, year={1999}, pages={681–687} }
 @article{engle_spears_brown_lloyd_1999, title={Effect of breed (Angus Vs Simmental) on immune function and response to a disease challenge in stressed steers and preweaned calves}, volume={77}, DOI={10.2527/1999.773516x}, abstractNote={Two experiments were conducted with feeder steer calves and preweaned calves to determine the effects of breed on immune response. In Exp. 1, newly weaned Angus (n = 24) and Simmental (n = 24) steer calves were blocked by weight within breed and randomly assigned to 12 pens with four calves per pen. The basal diet consisted of 87% corn silage (DM basis) and 13% of a soybean meal-mineral-vitamin supplement. Steers were allowed ad libitum access to feed throughout the study. On d 2 following weaning, calves received an intranasal inoculation of infectious bovine rhinotraecheitis virus (IBRV; 2.7 × 108 CCID50). Rectal temperatures in response to the IBRV were higher (P < .05) in Angus calves. On d 9, calves were injected i.m. with 10 mL of a 25% pig red blood cell (PRBC) suspension. Total immunoglobulin (Ig) and IgM titers against PRBC were higher (P < .05) for the Angus calves. Breed did affect cell-mediated immune response to phytohemagglutinin (PHA). In Exp. 2, preweaned (16 Angus and 16 Simmental) calves were selected based on breed, body weight, and sex. On 0 d, all selected calves were injected i.m. with 10 mL of a 25% PRBC suspension. Total Ig and IgG titers against PRBC were higher (P < .05) for Angus calves. On d 28, lymphocytes were isolated from peripheral blood obtained from eight calves per breed. Peripheral lymphocytes from the Angus calves had a greater (P < .07) blastogenic response to 6.25 µ/mL of PHA than lymphocytes from Simmental calves. Results indicate that the immune response of Angus and Simmental calves may differ.}, number={3}, journal={Journal of Animal Science}, author={Engle, T. E. and Spears, J. W. and Brown, T. T. and Lloyd, K. E.}, year={1999}, pages={516–521} }
 @article{walker_ahmed_brown_ho_hodges_lucier_russo_weigel_weise_vandenbergh_1999, title={Species, interindividual, and tissue specificity in endocrine signaling}, volume={107}, DOI={10.1289/ehp.99107s4619}, abstractNote={The activity of endocrine-active agents exhibits specificity at many levels. Differential responsiveness to these agents has been observed between different species and extends to interindividual differences within a species and between different tissues as well. In cases where they have been identified, the biologic and molecular mechanisms underlying this specificity are quite diverse. Determinants of species specificity include differences that exist in receptor binding, gene transcription, and cellular responses to endocrine-active compounds between species. Interindividual differences in responsiveness may be determined at the level of genetic polymorphisms in hormone-metabolizing enzymes, hormone receptors, and in those genes that are transactivated by these receptors, as well as during changing windows of susceptibility that occur as a function of age, such as prenatal and postmenopausal exposures. Extrinsic factors such as diet can also impact individual susceptibility to endocrine-active agents. Tissue-specific determinants of susceptibility are well documented, but little is known regarding the mechanisms underlying these different responses. Differences in the expression of accessory proteins for steroid hormone receptors and different patterns of receptor expression, estrogen receptor alpha and estrogen receptor beta; for example, may contribute to tissue specificity, as may differences in the pattern of expression of other genes such as hormone-metabolizing enzymes. The use of animal model systems and development of appropriate mathematical models has the potential to yield additional valuable information for elucidating the role of these determinants of specificity at low-dose exposures and for improved risk assessments for the adverse health effects of endocrine-active compounds.}, number={1999 Aug.}, journal={Environmental Health Perspectives}, author={Walker, C. and Ahmed, S. A. and Brown, T. and Ho, S. M. and Hodges, L. and Lucier, G. and Russo, J. and Weigel, N. and Weise, T. and Vandenbergh, J.}, year={1999}, pages={619–624} }
 @article{fernandez_grindem_brown_sharp_saulnier_1997, title={Cytologic and histologic features of a poorly differentiated glioma in a dog}, volume={26}, ISSN={["1939-165X"]}, DOI={10.1111/j.1939-165X.1997.tb00733.x}, abstractNote={A 5-year old female Boxer with a 1-week history of progressive paresis and paraplegia had a T10-13 subarachnoid filling defect on myelography. Exploratory hemilaminectomy revealed an intramedullary spinal cord tumor which was subsequently diagnosed as a poorly differentiated glioma, most likely an anaplastic ependymoma. The cytologic, histologic, and immunocytochemical staining characteristics of this neoplasm are described. Differential diagnoses, including primary and secondary tumors involving the central nervous system are discussed.}, number={4}, journal={VETERINARY CLINICAL PATHOLOGY}, author={Fernandez, FR and Grindem, CB and Brown, TT and Sharp, NJH and Saulnier, M}, year={1997}, pages={182–186} }
 @article{kegley_spears_brown_1997, title={Effect of shipping and chromium supplementation on performance, immune response, and disease resistance of steers}, volume={75}, DOI={10.2527/1997.7571956x}, abstractNote={Forty-eight Angus crossbred steers (263 ± 2 kg initial BW) were blocked by weight and randomly assigned within weight group to treatment. Treatments consisted of control or .4 mg of supplemental Cr as Cr-nicotinic acid complex/kg of DM. Steers were fed diets containing 90% corn silage (DM basis) and 10% of a soybean meal-mineral-vitamin supplement. After 56 d on the dietary treatment, half of the steers in each treatment were transported 343 km and unloaded in an unfamiliar location. The next day, d 58, shipped steers were returned to the feedlot (50 km). On d 58 after shipped steers were returned to the feedlot, all steers were inoculated with infectious bovine rhinotracheitis virus (IBRV) intranasally. Average daily gain from d 0 to 80 was increased (P < .10) by supplemental Cr. There was a shipping × time interaction for serum cortisol concentrations. Shipping increased (P < .02) serum cortisol on d 58, but 7 d after transport there were no effects of shipping on serum cortisol. Transportation increased (P < .05) the ratio of neutrophils to lymphocytes. Supplemental Cr did not affect rectal temperature after the IBRV challenge or the antibody response to IBRV or porcine red blood cells. Immunoglobulin G antibody response to porcine red blood cells was decreased (P < .09) by shipping. Supplemental Cr as Cr-nicotinic acid improved ADG of growing steers, regardless of whether they had been stressed by shipping. Supplemental Cr did not affect any of the immune responses that were measured.}, number={7}, journal={Journal of Animal Science}, author={Kegley, E. B. and Spears, J. W. and Brown, T. T.}, year={1997}, pages={1956–1964} }
 @article{gengelbach_ward_spears_brown_1997, title={Effects of copper deficiency and copper deficiency coupled with high dietary iron or molybdenum on phagocytic cell function and response of calves to a respiratory disease challenge}, volume={75}, DOI={10.2527/1997.7541112x}, abstractNote={A study was conducted to determine the effects of supplementing a diet marginally deficient in copper (Cu) with iron (Fe), molybdenum (Mo), or Cu on phagocytic cell function and disease resistance of calves. Thirty-one calves were born to heifers fed a corn silage-based diet containing 4.5 mg of Cu/kg. Treatments consisted of 1) control (CON; no supplemental Cu, Fe, or Mo), 2) 600 mg of Fe added/kg (FE), 3) 5 mg of Mo added/kg (MO), or 4) 10 mg of Cu added/kg of DM (CU). Activity of superoxide dismutase was lower (P < .06) in neutrophils from MO vs CON or CU calves at 170 d of age. bactericidal activity of neutrophils from MO calves tended (P = .15) to be lower compared with those from CU calves at 70 d of age. Calves were inoculated intranasally with live infectious bovine rhinotracheitis virus (IBRV) 2 d after weaning, followed by intratracheal administration of Pasteurella hemolytica 5 d later. Iron- and Cu-supplemented calves exhibited higher (P < .01) body temperatures and lower (P < .06) feed intakes following IBRV inoculation compared with CON and MO calves. Copper-supplemented calves had higher levels of plasma tumor necrosis factor (TNF) than MO calves at weaning (P < .05) and tended to have higher plasma TNF (P = .11) than FE and MO calves 5 d after IBRV inoculation. These data indicate that dietary levels of Mo and Cu can affect body temperature and feed intake responses to disease by affecting TNF and perhaps other cytokines.}, number={4}, journal={Journal of Animal Science}, author={Gengelbach, G. P. and Ward, J. D. and Spears, J. W. and Brown, T.T.}, year={1997}, pages={1112–1118} }
 @article{ritchey_degernes_brown_1997, title={Exocrine pancreatic insufficiency in a yellow-naped Amazon (Amazona ochrocephala) with pancreatic adenocarcinoma}, volume={34}, ISSN={["0300-9858"]}, DOI={10.1177/030098589703400111}, abstractNote={This report describes exocrine pancreatic insufficiency in a yellow-naped Amazon ( Amazona ochrocephala) with complete effacement of the pancreas by a pancreatic adenocarcinoma. The bird presented with a 3-month history of weight loss and voluminous, foul-smelling droppings. Clinically, routine hematologic findings were normal and fecal tests were performed to evaluate exocrine pancreatic function. The fecal function tests were positive for neutral and split fats and negative for trypsin. Oral administration of corn oil did not result in elevation of blood triglyceride levels. Two days later, the triglyceride tolerance test was repeated using corn oil mixed with pancreatic enzymes. This time, there was a 70% elevation of blood triglyceride levels. Because of a poor prognosis, the bird was euthanatized. At necropsy, the pancreas was diffusely enlarged, white, nodular, and firm. The liver contained multiple, 1-2-mm-diameter, randomly located, tan nodules. Microscopically, the pancreas was effaced by numerous lobules of neoplastic ductular structures surrounded by abundant fibrous connective tissue. In the liver, the hepatic parenchyma was replaced by multiple, well-demarcated, nonencapsulated foci of neoplastic tissue similar to that in the pancreas.}, number={1}, journal={VETERINARY PATHOLOGY}, author={Ritchey, JW and Degernes, LA and Brown, TT}, year={1997}, month={Jan}, pages={55–57} }
 @article{brown_shin_fuller_1995, title={Detection of pseudorabies viral DNA in tonsillar epithelial cells of latently infected pigs}, volume={56}, number={5}, journal={American Journal of Veterinary Research}, author={Brown, T. T. and Shin, K. O. and Fuller, F. J.}, year={1995}, pages={587} }
 @article{brown_shin_1990, title={Effect of bovine herpesvirus-1 or parainfluenza-3 virus on immune receptor-mediated functions of bovine alveolar macrophages in the presence or absence of virus-specific serum or pulmonary lavage fluids collected after virus infection}, volume={51}, number={10}, journal={American Journal of Veterinary Research}, author={Brown, T. T., Jr. and Shin, K.}, year={1990}, pages={1616} }
 @article{carver_shymodjeska_brown_rogers_riviere_1985, title={DOSE-RESPONSE STUDIES OF GENTAMICIN-NEPHROTOXICITY IN RATS WITH EXPERIMENTAL RENAL DYSFUNCTION .1. SUBTOTAL SURGICAL NEPHRECTOMY}, volume={80}, ISSN={["1096-0333"]}, DOI={10.1016/0041-008X(85)90082-1}, abstractNote={Gentamicin pharmacokinetics and nephrotoxicity have not been widely studied in animals with preexisting renal dysfunction, despite the fact that nephrotoxicity is a continuing manifestation of clinical therapy. The present study contrasted the dose-response nephrotoxicity of gentamicin in control rats with that of rats with renal insufficiency secondary to subtotal (2/3) surgical nephrectomy. Total daily doses ranging from 0 to 120 mg/kg were given in a divided regimen, every 8 hr and doses were reduced by doubling the interval in subtotally nephrectomized (Nx) rats, in proportion to impaired renal elimination on the first day of gentamicin administration. Estimates of renal function, including creatinine clearance, fractional sodium and potassium excretion, and serum creatinine and urea nitrogen, were collected after 6 and 12 days of dosing. In addition, urinary N-acetyl-beta-D-glucosaminidase excretion (6 days), in vitro renal cortical slice accumulation of tetraethylammonium (TEA) (6 days), quantified morphological lesions (12 days), and renal gentamicin concentrations (6 days) were examined. Pharmacokinetic data collected immediately after the first dose revealed a reduced gentamicin clearance and slightly reduced volume of distribution, with a corresponding prolonged half-life in the Nx rats. Based on statistical analysis of the dose-response relationships, Nx rats were functionally resistant to gentamicin nephrotoxicity after 6 days of dosing. This resistance was partially reversed by 12 days dosing, despite light-microscopic evidence of greater structural damage in the control rats. Renal parenchymal gentamicin concentrations were lower at some doses in the Nx rats, in contrast to the higher fractional reabsorption found in these rats at all doses. TEA transport was depressed at all doses in control rats but not at the lower doses in Nx rats, indicating that resistance was partially mediated at the level of the proximal tubular epithelium. This study demonstrates altered gentamicin pharmacokinetics and nephrotoxicity in a surgical model of renal dysfunction in rats.}, number={2}, journal={TOXICOLOGY AND APPLIED PHARMACOLOGY}, author={CARVER, MP and SHYMODJESKA, JS and BROWN, TT and ROGERS, RA and RIVIERE, JE}, year={1985}, pages={251–263} }