Duration of Mydriasis Produced by 0.5% and 1% Tropicamide in Sprague-Dawley Rats.

Ophthalmic examination (OE) is a common part of preclinical studies. Pupillary dilation to facilitate OE may affect results of other planned testing. The purpose of this study was to determine the duration of mydriasis produced by commercially available 0.5% and 1% tropicamide in ophthalmologically normal albino rats. Twelve female Sprague-Dawley rats were used. A single drop of 1% tropicamide was applied to one eye of each rat. A single drop of balanced salt solution (BSS) was applied to the contralateral eye. Measurements of pupillary diameter (PD) were obtained using a digital caliper at 0, 20, 40, 60, 120, 180, 240, 300, and 360 min after application. After a 3-wk washout period, the procedure was repeated using 0.5% tropicamide. Pupillary dilation sufficient to allow posterior segment evaluation was achieved with 0.5% and 1% tropicamide. Maximum PD after treatment with 0.5% tropicamide was 4.17 ± 0.22 mm at 40 min; maximum PD after treatment with 1% tropicamide was not significantly different (4.28 mm at both 20 and 40 min (±0.43 mm and 0.23 mm, respectively)). Mean PD remained above 3.5 mm in treated eyes for 60 min. In eyes treated with 0.5% tropicamide, mean PD was significantly different from baseline mean PD for that eye up to 300 min. In eyes treated with 1% tropicamide, mean PD was significantly different than baseline mean PD for that eye at all timepoints. Both concentrations of tropicamide produced a transient mild to moderate mydriasis in the contralateral eye. Duration of action is at least 5 h for 0.5% tropicamide and 6 h for 1% tropicamide. Results of this study support use of 0.5% tropicamide for OE in albino rats, with administration performed no more than 60 min prior to examination.

Spontaneous chronic T-cell leukemia in a male rhesus macaque ( Macaca mulatta).

Blood smears from a 24-year-old male rhesus macaque ( Macaca mulatta) used for cognitive function studies were evaluated. The macaque had an 8-month history of gradual weight loss and increasing lymphocytosis. Most of the lymphocytes present were small to medium and had a mature morphology. Based on the degree and duration of the lymphocytosis, and the appearance of the lymphocytes, a diagnosis of chronic lymphocytic leukemia was made. The animal tested negative for 4 viral diseases that are commonly associated with lymphoproliferative disorders in Old World monkeys. Over the course of 12 months, the lymphocytosis progressed from 18.4 to 384 × 103 lymphocytes/µl (reference range: 0.8-17 × 103 cells/µl), and euthanasia was elected. On histologic examination, cluster of differentiation (CD)3- and CD8-positive, CD79-negative neoplastic cells comprised 40-60% of the bone marrow, diffusely obscured the normal splenic architecture, and were present in the vascular channels in other organs. Findings were characteristic of T-cell lymphocytic leukemia. Naturally occurring T-cell lymphocytic leukemia has been rarely reported in rhesus macaques and, to the authors' knowledge, never in males. A persistent lymphocytosis characterized by a monomorphic population of CD3- and CD8-positive cytotoxic T-lymphocytes and the presence of neoplastic cells in the bone marrow led to a diagnosis in the current case.

Characterization of the natural history of extracellular matrix production in tissue-engineered vascular grafts during neovessel formation.

BACKGROUND: The extracellular matrix (ECM) is a critical determinant of neovessel integrity. MATERIALS AND METHODS: Thirty-six (polyglycolic acid + polycaprolactone and poly lactic acid) tissue-engineered vascular grafts seeded with syngeneic bone marrow mononuclear cells were implanted as inferior vena cava interposition grafts in C57BL/6 mice. Specimens were characterized using immunohistochemical staining and qPCR for representative ECM components in addition to matrix metalloproteinases (MMPs). Total collagen, elastin, and glycosaminoglycan (GAG) contents were determined. MMP activity was measured using zymography. RESULTS: Collagen production on histology demonstrated an initial increase in type III at 1 week followed by type I production at 2 weeks and type IV at 4 weeks. Gene expression of both type I and type III peaked at 2 weeks, whereas type IV continued to increase over the 4-week period. Histology demonstrated fibrillin-1 deposition at 1 week followed by elastin production at 4 weeks. Elastin gene expression significantly increased at 4 weeks, whereas fibrillin-1 decreased at 4 weeks. GAG demonstrated abundant production at each time point on histology. Gene expression of decorin significantly increased at 4 weeks, whereas versican decreased over time. Biochemical analysis showed that total collagen production was greatest at 2 weeks, and there was a significant increase in elastin and GAG production at 4 weeks. Histological characterization of MMPs showed abundant production of MMP-2 at each time point, while MMP-9 decreased over the 4-week period. Gene expression of MMP-2 significantly increased at 4 weeks, whereas MMP-9 significantly decreased at 4 weeks. CONCLUSIONS: ECM production during neovessel formation is characterized by early ECM deposition followed by extensive remodeling.

Fructosamine reference ranges in rhesus macaques (Macaca mulatta).

Naturally occurring diabetes mellitus (DM) is common in several species of Old and New World nonhuman primates. Fructosamine values provide important information about recent glycemic control and can be useful in the diagnosis and management of DM. However, despite an abundance of reports in the literature describing spontaneous and induced DM in monkeys, few reference ranges are available for fructosamine. Reference ranges have been published for woolly monkeys (Lagothrix lagotricha), cynomolgus macaques (Macaca fascicularis), and stumptail macaques (Macaca arctoides) but currently are not available for rhesus macaques. At our institution, DM is a common diagnosis in aging rhesus macaques. Here we report a reference range for fructosamine in rhesus macaques. The overall range was 157 to 230 µmol/L, with male rhesus and macaques 10 y or older having significantly higher values than do female rhesus and macaques younger than 10 y, respectively. This range provides clinical veterinarians with an additional tool for evaluating glycemic control in rhesus macaques.

Maropitant citrate for treatment of ulcerative dermatitis in mice with a C57BL/6 background.

Murine ulcerative dermatitis (UD) is a common progressive condition of mice with a C57BL/6 background. Typically, mice present with scabs and crusts on the skin of the dorsal neck and ears, and are often severely pruritic. Animals tend to scratch the lesions, causing additional trauma to the already ulcerated and inflamed skin. Therapeutic intervention largely has been unsuccessful, in part due to the lack of a known cause for the disease. Though the exact etiology of UD has not been elucidated, substance P (SP) has recently been demonstrated as an important neuropeptide linked to the itch-scratch cycle. SP functions at the tachykinin neurokinin 1 (NK1) receptor. We hypothesized that inhibition of SP binding to the NK1 receptor would decrease the itch sensation, thus decreasing scratching behavior and subsequent skin trauma. The purpose of this study was to evaluate the effectiveness of an NK1 receptor antagonist, maropitant citrate, as a treatment for murine UD. Treatment with 1 mg/kg maropitant citrate significantly reduced the size of UD lesions in mice.

Use of p63, a myoepithelial cell marker, in determining the invasiveness of spontaneous mammary neoplasia in a rhesus macaque (Macaca mulatta).

Here we describe a case of mammary gland ductal carcinoma in an aged rhesus macaque. Tumors were diagnosed based on routine hematoxylin and eosin staining. Invasiveness was further characterized by p63 immunohistochemistry. p63 is a p53 homolog that strongly and specifically stains nuclei of myoepithelial cells in human and canine mammary tissue. Because p63 has an affinity for the nucleus of myoepithelial cells, it is readily visible. Staining of mammary tissue from the monkey for p63 revealed that multiple foci of neoplastic cells had breached the myoepithelial cell layer surrounding ducts, suggesting the potential for local invasion of the tumor. Regional metastasis was confirmed at necropsy. To our knowledge, this is the first documented use of p63 for effectively determining the invasive nature of a mammary tumor in a nonhuman primate and the first use of p63 as an effective means of staining myoepithelial cells in a mammary ductal carcinoma in a nonhuman primate. Because nonhuman primates are important animal models for human diseases, including neoplasia, this method may prove useful for both diagnostic and research purposes.

Disseminated, histologically confirmed Cryptococcus spp infection in a domestic ferret.

CASE DESCRIPTION: A 4-year-old castrated male domestic ferret from central Massachusetts was evaluated for weight loss over a 1.5-month period and for 2 days of retching, diarrhea, and signs of lethargy. It had been housed indoors, with 2 other ferrets, 2 cats, and humans that lacked signs or symptoms of disease. CLINICAL FINDINGS: Physical examination revealed a thin body condition, tachypnea, an increase in respiratory effort, and retching. Splenomegaly was detected during abdominal palpation. Clinicopathologic analysis revealed lymphopenia, lactic acidosis, hypoglycemia, hypocalcemia, hypoalbuminemia, and hyperglobulinemia. A pulmonary bronchointerstitial pattern was evident on radiographs, and abdominal ultrasonography revealed a suspected pancreatic mass and mesenteric lymphadenopathy. TREATMENT AND OUTCOME: After 2 weeks of medical treatment and once clinical signs resolved, an exploratory laparotomy was performed and a lymph node biopsy specimen was collected. Histologic evaluation of the specimen revealed Cryptococcus-like organisms. Antifungal treatment was initiated with itraconazole (PO) and amphotericin B (IV). The ferret died after 2 days of treatment. A full necropsy was performed, revealing multicentric cryptococcosis affecting the lungs, brain, spleen, and multiple lymph nodes. Paraffin-embedded, formalin-fixed lung tissue was submitted for DNA extraction, and the organism was identified as Cryptococcus neoformans var grubii. CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of disseminated cryptococcosis in a North American ferret. This case is unique in that the ferret lived indoors, in a geographic region in which reports of cryptococcosis are rare. The genotyping technique used to identify the Cryptococcus strain can aid in better understanding the epidemiology of cryptococcosis.