Light-microscopy evaluation of zonular fiber morphology in dogs with glaucoma: secondary to lens displacement

Lens displacement resulting in secondary glaucoma is common in terrier breeds. This study was carried out to evaluate whether light microscopy was useful in detecting abnormal patterns in zonular fiber protein. Eyes from 63 dogs with glaucoma secondary to lens displacement were evaluated for the presence of abnormal zonular fiber morphology using the following stains: hematoxylin and eosin, periodic acid Schiff (PAS), Masson's trichrome and Verhoeff's elastin stains. Two distinct forms of abnormal zonular fiber morphology were recognized and designated as zonular fiber dysplasia (ZFD) and zonular fiber collagenization (ZFC). ZFD protein morphology was characterized by being tightly adherent to the nonpigmented ciliary body epithelium, exhibiting a distinct lamellar and cross-hatched pattern and staining strongly positive with PAS and trichrome stains, and staining negative with elastin stains. ZFD was predominant in terrier breeds (18 of 29) and Shar-Pei dogs (4 of 29). ZFC abnormality was characterized by excessive zonular fiber that was not tightly adherent to the ciliary body epithelium and staining positive with PAS, trichrome (blue for collagen) and elastin stains. Only 7 of 19 dogs with ZFC changes were terrier breeds, and there was no pattern in the breeds affected. Fifteen of the 63 dogs used in the study had normal appearing zonular fibers. The staining pattern in these dogs matched normal controls by staining positive with PAS and Verhoeff's elastin stains and had only minimal positive staining with Masson's trichrome stain. Results suggest that light microscopy is useful in detecting breed-related changes in zonular fiber morphology in cases of glaucoma secondary to lens displacement. These changes may correlate with the presence of abnormal zonular fiber proteins and might be important in the pathogenesis of primary lens displacement in terrier and Shar-Pei dogs.     

宫内生长迟缓仔猪T细胞的发育研究

研究旨在探讨从出生到断奶不同日龄宫内生长迟缓(IUGR)猪T细胞的发育变化情况。选用体重相近和配种期相同的达兰初产妊娠母猪30头,分娩时从每窝仔猪中挑选1头正常体重猪和1头IUGR猪,分别在1、7、14、21、28日龄时屠宰取胸腺,分离T细胞,通过流式细胞仪测定CD3、CD4、CD8T细胞的数量,每个日龄6个重复。结果表明:出生后第1天IUGR仔猪与正常仔猪的CD4+CD8+细胞(双阳性T细胞)占总T细胞的百分比差异显著(P<0.05);出生后第7天IUGR仔猪与正常仔猪的CD4细胞占总T细胞的百分比差异极显著(P<0.01);后期各细胞亚群之间没有差异。结果提示,子宫内各种因素的综合影响能造成出生后早期IUGR仔猪T细胞亚群发育不完善,这就从T细胞亚群的发育方面解释了IUGR仔猪免疫力低的原因。     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

用直接技术测定玉米蛋白粉猪回肠表观氨基酸消化率

本试验采用直接技术 ,测定玉米蛋白粉的 18种猪回肠表观氨基酸消化率。选择的玉米蛋白粉是具有我国代表性的玉米加工副产品 ,粗蛋白质含量分别为 6 3% ,5 2 %和 4 7%。 6头T型瘘管猪平均体重 (4 7± 2 .81)kg,6× 6拉丁方试验设计。试验结果表明 ,玉米蛋白粉的蛋氨酸、酪氨酸、亮氨酸和苯丙氨酸回肠表观氨基酸消化率相对较高 ;不同蛋白质含量的玉米蛋白粉的色氨酸、酪氨酸和亮氨酸回肠表观消化率有显著性差异 (P <0 .0 5 )。赖氨酸、色氨酸和组氨酸分别是玉米蛋白粉的第一、第二和第三限制性氨基酸。加工工艺影响玉米蛋白粉猪回肠表观氨基酸消化率     

Risk factors and prevalence of injuries in horses during various types of steeplechase races

OBJECTIVE: To identify the types of injuries sustained by horses that competed in steeplechase races and determine the prevalence of and risk factors for those injuries. DESIGN: Prospective study. ANIMALS: 2,680 horses that competed in various types of steeplechase races from 1996 through 2000. PROCEDURE: Data regarding races; environment; equipment problems; the number of horses that entered, started, and finished races; the number of riders that fell; and the number of horses that were slowed or stopped by the rider, ran off the course, fell, and sustained injuries or physical abnormalities during races were collected on a standard form by the official veterinarian who attended each meet. Data from all meets were not recorded; however, in recorded meets, data from every race were reported. RESULTS: Data for 197 hurdle, 65 timber, 76 flat, and 8 mixed races were recorded. Nine (3.4/1,000 horses that started in races) horses died or were euthanatized, and 7 of those were associated with catastrophic musculoskeletal injury. Seven fractures were recorded. Four fractures involved forelimbs, 1 involved a hind limb, and 2 involved the cervical portion of the vertebral column. All horses with fractures were euthanatized. Deep or hard course conditions were associated with an increased risk of breakdown injuries. CONCLUSIONS AND CLINICAL RELEVANCE: Successful development and implementation of strategies to prevent injuries and death in horses in steeplechase races depend on a clear understanding of the types and prevalence of injuries involved and risk factors associated with those injuries.     

Leptin mRNA expression and serum leptin concentrations as influenced by age, weight, and estradiol in pigs

Two experiments (EXP) were conducted to determine the roles of age, weight and estradiol (E) treatment on serum leptin concentrations and leptin gene expression. In EXP I, jugular blood samples were collected from gilts at 42 to 49 (n = 8), 105 to 112 (n = 8) and 140 to 154 (n = 8) d of age. Serum leptin concentrations increased (P < 0.05) with age and averaged 0.66, 2.7, and 3.0 ng/ml (pooled SE 0.21) for the 42- to 49-, 105- to 112-, and 140- to 154-d-old gilts, respectively. In EXP II, RNase protection assays were used to assess leptin mRNA in adipose tissue of ovariectomized gilts at 90 (n = 12), 150 (n = 11) or 210 (n = 12) d of age. Six pigs from each age group received estradiol (E) osmotic pump implants and the remaining animals received vehicle control implants (C; Day 0). On Day 7, back fat and blood samples were collected. Estradiol treatment resulted in greater (P < 0.05) serum E levels in E (9 +/- 1 pg/ml) than C (3 +/- 1 pg/ml) pigs. Serum leptin concentrations were not affected by age, nor E treatment. Leptin mRNA expression was not increased by age in C pigs nor by F in 90- and 150-d-old pigs. However, by 210 d of age, leptin mRNA expression was 2.5-fold greater (P < 0.01) in E-treated pigs compared to C animals. Serum insulin concentrations were similar between treatments for 210-d-old pigs. However, insulin concentrations were greater (P < 0.05) in E than C pigs at 90 d and greater in C than E animals at 150 d. Plasma glucose and serum insulin-like growth factor-I concentrations were not influenced by treatment. These results demonstrate that serum leptin concentrations increased with age and E-induced leptin mRNA expression is age- and weight-dependent.     

Congenital stationary night blindness is associated with the leopard complex in the Miniature Horse

Objective To determine if congenital stationary night blindness (CSNB) exists in the miniature horse in association with leopard complex spotting patterns (LP), and to investigate if CSNB in the miniature horse is associated with three single nucleotide polymorphisms (SNPs) in the region of TRPM1 that are highly associated with CSNB and LP in Appaloosas. Animals studied Three groups of miniature horses were studied based on coat patterns suggestive of LP/LP (n = 3), LP/lp (n = 4), and lp/lp genotype (n = 4). Procedures Horses were categorized based on phenotype as well as pedigree analysis as LP/LP, LP/lp, and lp/lp. Neurophthalmic examination, slit‐lamp biomicroscopy, indirect ophthalmoscopy, and scotopic flash electroretinography were performed on all horses. Hair samples were processed for DNA analysis. Three SNPs identified and associated with LP and CSNB in the Appaloosa were investigated for association with LP and CSNB in these Miniature horses. Results All horses in the LP/LP group were affected by CSNB, while none in the LP/lp or lp/lp groups were affected. All three SNPs were completely associated with LP genotype (χ² = 22, P << 0.0005) and CSNB status (χ² = 11, P < 0.0005). Conclusions The Miniature Horse breed is affected by CSNB and it appears to be associated with LP as in the Appaloosa breed. The SNPs tested could be used as a DNA test for CSNB until the causative mutation is determined.     

Canine corneal fibroblast and myofibroblast transduction with AAV5

Objective The aims of this study were (1) to determine the efficacy of adeno‐associated vector serotype 5 (AAV5) for delivering gene therapy to canine corneal fibroblasts (CCFs) and myofibroblasts (CCMs) using enhanced green fluorescent protein (GFP) marker gene and (2) to evaluate the cytotoxicity of AAV5 to CCFs and CCMs using an in vitro model. Methods Healthy donor canine corneas were used to generate primary CCFs by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). An AAV5 titer (6.5 × 10¹² μg/mL) expressing GFP under control of hybrid cytomegalovirus + chicken β‐actin promoters (AAV5‐gfp) was used to transduce CCF and CCM cultures. Delivered gene expression in CCFs and CCMs was quantified using immunocytochemistry, fluorescent microscopy, and real‐time PCR. Transduction efficacy of the AAV5 vector was determined by counting DAPI‐stained nuclei and EGFP‐positive cells in culture. Phase‐contrast microscopy, trypan blue, and dUTP nick end labeling (TUNEL) assays were used to determine the toxicity and safety of AAV5 in this canine corneal model. Results Topical AAV5 application successfully transduced a significant population of CCFs (42.8%; P < 0.01) and CCMs (28%; P < 0.01). Tested AAV5 did not affect CCF or CCM phenotype or cellular viability and did not cause significant cell death. Conclusions The tested AAV5 is an effective and safe vector for canine corneal gene therapy in this in vitro model. In vivo studies are warranted.     

Diabetes mellitus-related morphoquantitative changes in the celiac ganglion neurons of the dog

Diabetes mellitus is the most common endocrine disturbance of domestic carnivores and can cause autonomic neurological disorders, although these are still poorly understood in veterinary medicine. There is little information available on the quantitative adaptation mechanisms of the sympathetic ganglia during diabetes mellitus in domestic mammals. By combining morphometric methods and NADPH-diaphorase staining (as a possible marker for nitric oxide producing neurons), type I diabetes mellitus-related morphoquantitative changes were investigated in the celiac ganglion neurons in dogs. Twelve left celiac ganglia from adult female German shepherd dogs were examined: six ganglia were from non-diabetic and six from diabetic subjects. Consistent hypertrophy of the ganglia was noted in diabetic animals with increase of 55% in length, 53% in width, and 61.5% in thickness. The ordinary microstructure of the ganglia was modified leading to an uneven distribution of the ganglionic units and a more evident distribution of axon fascicles. In contrast to non-diabetic dogs, there was a lack of NADPH-diaphorase perikarial labelling in the celiac ganglion neurons of diabetic animals. The morphometric study showed that both the neuronal and nuclear sizes were significantly larger in diabetic dogs (1.3 and 1.39 times, respectively). The profile density and area fraction of NADPH-diaphorase-reactive celiac ganglion neurons were significantly larger (1.35 and 1.48 times, respectively) in non-diabetic dogs compared to NADPH-diaphorase-non-reactive celiac ganglion neurons in diabetic dogs. Although this study suggests that diabetic neuropathy is associated with neuronal hypertrophy, controversy remains over the possibility of ongoing neuronal loss and the functional interrelationship between them. It is unclear whether neuronal hypertrophy could be a compensation mechanism for a putative neuronal loss during the diabetes mellitus.