This study was planned to determine the effect of certain vitamin applications on antioxidant and oxidant parameters in the osteoblast cell line exposed to sodium fluoride in vitro and to evaluate the protective role of certain vitamins against possible toxic effects of fluoride. Cells were replicated in vitro conditions with regular passaging 2-3 times weekly. MTF viability test was used to determine IC50 of NaF (5000μM) and proliferative doses of vitamins (Vitamin A: 10μM, Vitamin D: 10μM, Vitamin E: 60μM, Vitamin C: 100μM) for hFOB 1.19 cells. Cells were sown in flasks as so to be 106. The study groups were identified as control, NaF, vitamins and NaF+vitamins. After incubation for 24 hours, cells treated with trypsin were prepared by freeze/thaw method and MTT viability test, TAS, SOD, GSH, CAT, TOS and MDA analyzes were performed on these samples.In the hFOB 1.19 cell line, TAS levels decreased significantly in the NaF group (p≤0.05), but were close to the control group in NaF+vitamin groups with the exception of vitamin C. However, there was no difference between the groups in terms of GSH level and CAT and SOD activities when the control and NaF groups were compared. It was observed that TOS level increased significantly in the NaF group (p<0.05), decreased in the NaF+vitamin groups and were lower in the NaF+vitamin C and E groups than the control group (p <0.05). While OSI was the highest in the NaF group, no significant difference in MDA level was observed compared with the control group.Conclusion: As a result, it was found that NaF administration in the osteoblast cell line increased oxidative stress and decreased following vitamin application. It was found that the effect of NaF administration in the osteoblast cell line on cell viability was consistent with the oxidative stability and that the vitamin application conformably changed cell viability and oxidative balance.

8OBJECTIVE To develop a method to maintain the initial phenotype of airway smooth muscle (ASM) cells isolated from equine endobronchial biopsy specimens in long-term cell culture. SAMPLE Endobronchial tissue specimens (8 to 10/horse) collected from the lungs of previously healthy horses at necropsy (n = 12) and endobronchial biopsy specimens collected from standing, sedated, heaves-affected horses in clinical remission of the disease (5) and control horses (4). PROCEDURES A sampling protocol was developed to recover and maintain a contractile phenotype in ASM cells from endobronchial specimens from freshly harvested equine lungs and from healthy and heaves-affected horses. Immunologic techniques were used to evaluate the contractile phenotype of ASM cells in culture. RESULTS Characteristic ASM cells were successfully cultured from endobronchial tissue or biopsy specimens from both healthy and heaves-affected horses, and their contractile phenotype was maintained for up to 7 passages. Moreover, the capacity of cells at the seventh passage to contract in a collagen gel in response to methacholine was maintained. CONCLUSIONS AND CLINICAL RELEVANCE ASM cells isolated from equine endobronchial tissue and biopsy specimens were able to maintain a contractile phenotype in long-term cell cultures, suggesting they could be used for tissue engineering and in vitro studies of equine ASM cells.

OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses. ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes. PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days −6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed. RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses. CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.