A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts.

With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell-cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen transmission that might arise from animal-derived materials.

Under-utilization of health care services for infectious diseases syndromes in rural Azerbaijan: a cross-sectional study.

BACKGROUND: Infectious diseases present a potentially substantial yet undefined burden on the health of the adult Azerbaijani population. Efforts to quantify this burden in Azerbaijan are currently based almost exclusively on passive disease surveillance, and therefore hinge on the health utilization practices of the population. Understanding the prevalence of infectious syndromes and health utilization practices is paramount to disease surveillance, public health planning, and health care system reform. METHODS: A two-stage, probability proportional to size sampling design was used to select a representative sample of three regions of northern Azerbaijan with village populations less than 500 people. Demographic, clinical, and epidemiologic parameters were assessed using prevalence odds ratios, chi-squared, and the Fisher exact test. Associations with p < 0.10 were included in the regression analysis and removed by backward elimination. Respondents included 796 adults from 39 villages. RESULTS: Self-medication with antibiotics was the predominant utilization practice reported (19.4%). Only 1.3% of respondents reported seeing a health care provider for an infection, and 3.4% missed work or stayed in bed during the day in the last 5 years. In contrast, 338 illness episodes were reported in a 5 year period. Antibiotic use was significantly associated with gender, region, history of febrile illness, sleep disturbances, and arthritis controlling for age, ethnicity, and education. Influenza-like illness was the most prevalent infectious syndrome reported (33.3%). CONCLUSIONS: We observed a remarkably low utilization of health services, despite reported symptoms that would merit use. Widespread availability of antibiotics may deter health care use, and may contribute to the development of antibiotic resistance in this population. Information on utilization of health services during an infection is essential for development of effective intervention strategies, and data on the prevalence of infectious syndromes provides information not otherwise available in populations with low health care utilization.

Seroprevalence of tularemia in rural Azerbaijan.

A representative, two-stage probability sampling design was used to select 40 villages in northern Azerbaijan with populations of <500 people to screen for evidence of prior infection with Francisella tularensis. Informed consent was provided, and samples were obtained from 796 volunteers and tested for the presence of immunoglobulin G antibodies using enzyme-linked immunosorbent assay. F. tularensis seropositivity was prevalent (15.5% of volunteers), but there was minimal reporting of signs and symptoms consistent with clinical tularemia, suggesting that mild or asymptomatic infection commonly occurs. Frequently seeing rodents around the home was a risk factor for seropositivity (POR = 1.6, p = 0.03), controlling for age and gender. Geospatial analysis identified associations between village-level tularemia prevalence and suitable tick habitats, annual rainfall, precipitation in the driest quarter, and altitude. This study contributes to the growing understanding of the geographic distribution of tularemia and provides further information on the climatic and landscape conditions that increased the potential for exposure to this pathogen. The potential occurrence of asymptomatic or mild F. tularensis infection warrants further study.