Introduction: Production of nitric oxide (NO) is one of the main responses elicited by a variety of immune cells such as macrophages (e.g. microglia, resident macrophages of brain), during inflammation. Evaluation of NO levels in the inflammatory milieu is considered important to the understanding of the intensity of an immune response; and has been performed using different methods including the Griess assay. To assay NO in culture, an appropriate number of cells are stimulated into an inflammatory phenotype. Common stimuli include lipopolysaccharide (LPS), IFN-γ and TNF-α. However, overt stimulation could cause cell cytotoxicity therefore an ideal concentration of LPS should be used. Objective: To set-up a model of BV-2 cell activation that allows the assay of detectable levels of NO. Optimization of BV-2 microglia cell density and LPS concentrations after stimulation by bacterial lipopolysaccharide (LPS) for the Griess assay is demonstrated in this study. Methods: BV-2 microglia were cultured at different cell densities, and treated with LPS at three concentrations (1, 5, 10 µg/ml). NO production in culture supernatants were then measured at 18, 24, 48 and 72 hours. Moreover, methyl tetrazolium assay (MTT) was also performed to ensure that NO measurement is performed at no-cytotoxic concentrations of LPS. Results and Conclusions: NO production follows a temporal pattern. The density of 25000 cells/ well was the ideal seeding density for NO evaluation in BV-2 cells. BV-2 stimulation by LPS is dose dependent, and NO levels are increased proportional to the LPS concentration up to 1.0µg/ml, whereas the higher LPS concentrations are associated with decreased cell viability may be caused by the high toxic levels of LPS or NO. Although Griess assay has been commonly used by the scientists, however, optimization of its parameters on BV-2 cells will be useful for the experiments which will be performed on this particular cell line. The optimized pattern of Griess assay on BV-2 cells was achieved in this study, hence easier and more practical for the future scientists to perform Griess assay on BV-2 cells.
Nitric Oxide

BACKGROUND: Musculoskeletal disorders (MSDs) are a common problem among carpet weavers. This study was undertaken to introduce affecting personal and occupational factors in developing the number of MSDs among carpet weavers. METHODS: A cross-sectional study was performed among 862 weavers in seven towns with regard to workhouse location in urban or rural regions. Data were collected by using questionnaires that contain personal, workplace, and information tools and the modified Nordic MSDs questionnaire. Statistical analysis was performed by applying Poisson and negative binomial mixed models using a full Bayesian hierarchical approach. The deviance information criterion was used for comparison between models and model selection. RESULTS: The majority of weavers (72%) were female and carpet weaving was the main job of 85.2% of workers. The negative binomial mixed model with lowest deviance information criterion was selected as the best model. The criteria showed the convergence of chains. Based on 95% Bayesian credible interval, the main job and weaving type variables statistically affected the number of MSDs, but variables age, sex, weaving comb, work experience, and carpet weaving looms were not significant. CONCLUSION: According to the results of this study, it can be concluded that occupational factors are associated with the number of MSDs developing among carpet weavers. Thus, using standard tools and decreasing hours of work per day can reduce frequency of MSDs among carpet weavers.
Animals ; Bayes Theorem ; Comb and Wattles ; Cross-Sectional Studies ; Female ; Floors and Floorcoverings* ; Humans ; Iran* ; Occupational Diseases ; Risk Factors*