Evaluation of electroacupuncture for symptom modification in a rodent model of spontaneous osteoarthritis.

OBJECTIVE: Faced with the frustration of chronic discomfort and restricted mobility due to osteoarthritis (OA), many individuals have turned to acupuncture for relief. However, the efficacy of acupuncture for OA is uncertain, as much of the evidence is inconclusive. The purpose of this study was to evaluate electroacupuncture (EA) in a rodent model of OA such that conclusions regarding its effectiveness for symptom or disease modification could be drawn. METHODS: Ten 12-month-old male Hartley guinea pigs-which characteristically have moderate to advanced OA at this age-were randomly assigned to receive EA for knee OA (n = 5) or anesthesia only (control group, n = 5). Treatments were performed three times weekly for 3 weeks, followed by euthanasia 2 weeks later. Gait analysis and enclosure monitoring were performed weekly to evaluate changes in movement. Serum was collected for inflammatory biomarker testing. Knee joints were collected for histology and gene expression. RESULTS: Animals receiving EA had significantly greater changes in movement parameters compared to those receiving anesthesia only. There was a tendency toward decreased serum protein concentrations of complement component 3 (C3) in the EA group compared to the control group. Structural and antioxidant gene transcripts in articular cartilage were increased by EA. There was no significant difference in total joint histology scores between groups. CONCLUSION: This study provides evidence that EA has a positive effect on symptom, but not disease, modification in a rodent model of OA. Further investigations into mechanistic pathways that may explain the efficacy of EA in this animal model are needed.

Cytologic features and staining characteristics of Gamna-Gandy bodies from seven canine fine-needle aspirate preparations.

Gamna-Gandy (GG) bodies are non-infectious, hyphal-like structures associated with siderotic nodules in lymphoid tissue; GG bodies are very rarely reported in veterinary cytologic samples. Cytologically, GG bodies can be misidentified as hyphae or plant material. Seven canine lymphoid tissue aspiration cases that contained GG bodies were investigated for morphologic variability and staining characteristics. Available archived cytology slides containing GG bodies were stained with reagents known to show positive results (Prussian blue, Alizarin red S, Von Kossa) and negative results (Gomori methenamine silver) in histologic samples. Calcofluor white staining was also performed. GG bodies in Wright-Giemsa-stained cytology samples displayed considerable variability but were generally 2-5 µm diameter, 10-35 µm long, refractile, clear, pale-tan or pale-yellow, wavy or straight, tubular structures. Six cases allowed for cytochemical staining; staining properties were similar to histology samples. The bodies did not stain with calcofluor white; this stain may be helpful in distinguishing GG bodes from fungal hyphae.

Canine bicavitary carcinomatosis with transient needle tract metastasis diagnosed by multiplex immunocytochemistry.

A 6-year-old, male castrated, mixed-breed dog was referred to the James L. Voss Veterinary Teaching Hospital at Colorado State University for bicavitary effusion. On examination, the dog was tachycardic and tachypneic with bilaterally decreased lung sounds. Thoracic and abdominal ultrasonic examination revealed pleural and peritoneal effusions, which were aspirated and submitted for fluid analysis and cytology. Both cavity fluids were classified as exudates with a large population of vacuolated mononuclear cells. Multiplex immunocytochemistry (ICC) for cytokeratin and vimentin demonstrated exclusively cytokeratin expression, indicating these cells were of epithelial origin. A full diagnostic evaluation was performed, including CBC, clinical chemistry, a pet-side test for heartworm disease, ehrlichiosis, Lyme disease, and anaplasmosis, imaging modalities of thorax, abdomen, and heart, urinalysis, and fine-needle aspirations of spleen, liver, and popliteal lymph nodes. The dog was diagnosed with pleural and peritoneal carcinoma with presumed carcinomatosis. A single dose of intracavitary carboplatin was administered before discharge, and over a period of 2 weeks, 5 thoracocenteses were performed. A subcutaneous mass was noted at a thoracocentesis site one week after initial presentation. Cytology of the mass was consistent with carcinoma, and neoplastic seeding of the tumor cells from the thoracocentesis was suspected. The dog was euthanized 15 days after the first visit, and a necropsy was performed. Findings were consistent with carcinomatosis secondary to anaplastic pulmonary carcinoma with transient subcutaneous seeding of neoplastic cells during routine thoracocentesis. This case demonstrates the utility of multiplex ICC in the clinical setting.

Prior mucosal exposure to heterologous cells alters the pathogenesis of cell-associated mucosal feline immunodeficiency virus challenge.

BACKGROUND: Several lines of research suggest that exposure to cellular material can alter the susceptibility to infection by HIV-1. Because sexual contact often includes exposure to cellular material, we hypothesized that repeated mucosal exposure to heterologous cells would induce an immune response that would alter the susceptibility to mucosal infection. Using the feline immunodeficiency virus (FIV) model of HIV-1 mucosal transmission, the cervicovaginal mucosa was exposed once weekly for 12 weeks to 5,000 heterologous cells or media (control) and then cats were vaginally challenged with cell-associated or cell-free FIV. RESULTS: Exposure to heterologous cells decreased the percentage of lymphocytes in the mucosal and systemic lymph nodes (LN) expressing L-selectin as well as the percentage of CD4+ CD25+ T cells. These shifts were associated with enhanced ex-vivo proliferative responses to heterologous cells. Following mucosal challenge with cell-associated, but not cell-free, FIV, proviral burden was reduced by 64% in cats previously exposed to heterologous cells as compared to media exposed controls. CONCLUSIONS: The pathogenesis and/or the threshold for mucosal infection by infected cells (but not cell-free virus) can be modulated by mucosal exposure to uninfected heterologous cells.

Analysis and cytologic characterization of hemocytes from freshwater mussels (Quadrula sp.).

BACKGROUND: Freshwater mussels are among the most endangered taxa in North America and minimally invasive techniques to evaluate their health are needed. OBJECTIVE: The objective of this study was to develop a standardized approach for identifying and enumerating the cellular components of freshwater mussel hemolymph. METHODS: Hemocyte clumping, total hemocyte count, and hemocyte morphology were compared in untreated hemolymph or hemolymph treated with formalin, sodium citrate, sodium heparin, EDTA, water, or l-cysteine. Morphology was then used to categorize hemocytes and perform a 100-cell differential. RESULTS: Treatment with formalin or >25 mg/mL l-cysteine reduced hemocyte clumping, although only formalin significantly increased the total hemocyte count. However, formalin also induced crenation that impaired hemocyte identification. Both EDTA and sodium citrate-induced hemocyte degranulation while sodium citrate and >40 mg/mL l-cysteine-induced cell lysis. Hemocytes could be categorized into 2 groups of granulocytes (eosinophilic or basophilic) and 2 groups of agranulocytes (large or small) for performing a cytologic differential. The differential was not significantly altered by anticoagulant treatments providing cell morphology was adequate for obtaining a differential. Eosinophilic granulocytes predominated (59%) with fewer large agranulocytes (27%) and basophilic granulocytes (13%). Small agranulocytes comprised 2% of the total population. CONCLUSIONS: No single treatment provided an optimal method to evaluate freshwater mussel hemolymph. Maximal hemocyte counts were obtained following formalin treatment. l-cysteine reduced clumping and maintained hemocyte morphology for performing a cytologic differential. These techniques provide a standardized approach for the hematologic evaluation of freshwater mussels.

Mucosal administration of low-dose cell-associated feline immunodeficiency virus promotes viral latency.

Human immunodeficiency virus type 1 can occasionally be detected as a cryptic or latent infection in seronegative, asymptomatic patients. To develop an animal model of host latency, cats were mucosally challenged with 10(2)-10(6) feline immunodeficiency virus (FIV)-infected T cells. Although high-dose exposure (10(4)-10(6) T cells) resulted in progressive infection, no evidence of infection was seen in 5 of 6 cats exposed to 10(2) or 10(3) T cells. However, after ex vivo CD8(+) T cell depletion and phorbol myristate acetate treatment, FIV could be reactivated in tissues from 4 cats. Thus, latent tissue viral reservoirs can be induced by low-dose cell-associated mucosal challenge, providing a model to dissect the mechanisms that control reservoir establishment.

Induction of serum and mucosal FIV-specific immune responses by intranasal immunization with p24Gag.

We examined the ability of FIV p24Gag to induce systemic and mucosal FIV-specific immune responses when delivered as a nasal immunogen alone, or with a mucosal adjuvant, Escherichia coli heat labile toxin LT(R192G). Nasal immunization with p24Gag alone induced FIV-specific immune responses but overall responses were weak, transient, and/or present only in a few animals. Co-administration of LT(R192G) resulted in strong FIV-specific serum IgG and enhanced salivary IgA responses. Moreover, FIV-specific IgA was detected in vaginal wash fluid from 6/6 cats co-immunized with LT(R192G) and p24Gag versus 1/6 immunized with p24Gag alone. This is the first report detailing induction of systemic or mucosal FIV-specific immune responses by nasal immunization alone. As such, this study demonstrates that nasal immunization of cats can be a relevant and effective route for the delivery of candidate vaccines. However, while nasal immunization of cats with p24Gag induces antigen-specific systemic immune responses, development of strong systemic and mucosal immune responses requires co-administration of a mucosal adjuvant, such as LT(R192G).

Evaluation of FIV protein-expressing VEE-replicon vaccine vectors in cats.

Venezuelan equine encephalitis (VEE) virus-replicon particles (VRP) were used to generate feline immunodeficiency virus (FIV) Gag- and ENV-expressing vaccine vectors. Serum and mucosal FIV-specific antibody was detected in cats immunized subcutaneously, once monthly for 5 months, with FIV-expressing VRP. Expansion of the CD8+ L-selectin negative phenotype and transient CD8+ noncytolytic suppressor activity were seen in cats immunized with FIV-expressing or control VRP. Despite induction of FIV-specific immune responses and nonspecific suppressor responses, all cats became infected following vaginal challenge with high dose, pathogenic cell-associated FIV-NCSU(1) although relative early maintenance of CD4+ cells was seen in FIV-immunized cats.