Evaluation of IL-12RB1, IL-12B, CXCR-3 and IL-17a expression in cases affected by a non-healing form of cutaneous leishmaniasis: an observational study design.

INTRODUCTION: Seldom cutaneous leishmaniasis (CL) may present as a lasting and active lesion(s), known as a non-healing form of CL (NHCL). Non-functional type 1 T helper (Th1) cells are assumed the most important factor in the outcome of the disease. The present study aims to assess some molecular defects that potentially contribute to Th1 impairment in NHCL. METHODS AND ANALYSIS: This prospective observational study will be implemented among five groups. The first and second groups comprise patients afflicted with non-healing and healing forms of CL, respectively. The third group consists of those recovered participants who have scars as a result of CL. Those participants who have never lived or travelled to endemic areas of leishmaniasis will comprise the fourth group. The fifth group comprises participants living in hyperendemic areas for leishmaniasis, although none of them have been afflicted by CL. The aim is to recruit 10 NHCL cases and 30 participants in each of the other groups. A leishmanin skin test (LST) will be performed to assess in vivo immunity against the Leishmania infection. The cytokine profile (interleukin (IL)-12p70, interferon (IFN)-γ, C-X-C motif chemokine ligand (CXCL)-11 and IL-17a) of the isolated peripheral blood mononuclear cells (PBMCs) will be evaluated through ELISA. Real-time PCR will determine the C-X-C motif chemokine receptor (CXCR)-3 and IL-17a gene expression and expression of IL-12Rß1 will be assessed by flow cytometry. Furthermore, IL-12B and IL-12RB1 mutation analysis will be performed. DISCUSSION: It is anticipated that the outcome of the current study will identify IL-12B and IL-12RB1 mutations, which lead to persistent lesions of CL. Furthermore, our expected results will reveal an association between NHCL and pro-inflammatory cytokines (IL-12p70, IFN-γ IL-17a and CXCL-11), as well as CXCR-3 expression. ETHICS AND DISSEMINATION: This study has been approved by a local ethical committee. The final results will be disseminated through peer-reviewed journals and scientific conferences.

Tolerance Induction of Temperature and Starvation with Tricalcium Phosphate on Preservation and Sporulation in Bacillus amyloliquefaciens Detected by Flow Cytometry.

The Bacillus species have many applications in the preparation of various enzymes, probiotic, biofertilizer, and biomarkers for which the survival of resting cells and spore formation under different conditions are important. In this study, water and saline along with different mineral substances such as calcium carbonate, calcium phosphate, and silica were used for the detection of survival and preservation of Bacillus amyloliquefaciens. The results showed intensive death of resting cells at 8 °C, but significant survival at 28 °C after one month. However, preservation by minerals significantly decreased the rate of death and induced sporulation at both the temperatures. The resting cells were maintained at room temperature (about 60 % of the initial population survived after a month) in the presence of tricalcium phosphate. The results showed that temperature has more effect on sporulation compare with starvation. The sporulation in normal saline at 28 °C was 70 times more than that at 8 °C; meanwhile, addition of tricalcium phosphate increases sporulation by 90 times. Also, the FTIR data showed the interaction of tricalcium phosphate with spores and resting cells. The discrimination of sporulation from non-sporulation state was performed by nucleic acid staining with thiazole orange and detected by flow cytometry. The flow cytometric studies confirmed that the rates of sporulation in pure water were significantly more at 28 °C. This is the first report on the detection of bacterial spore with thiazole orange by flow cytometry and also on the interaction of tricalcium phosphate with spores by FTIR analyses.

The effects of bare metal versus drug-eluting stent implantation on circulating endothelial cells following percutaneous coronary intervention.

BACKGROUND: The purpose of this study was to compare the effects of bare metal stents (BMS) and drug-eluting stents (DES) implantation on circulating endothelial cells (CECs) which have been proposed as cellular markers of endothelial dysfunction following percutaneous coronary intervention (PCI). Recently, it has been established that DES further reduce restenosis and revascularization rate compared to bare metal stents in elective procedures. However, its benefits are compromised by the stent-related thrombosis events. METHODS: 22 patients who were candidate of PCI were included in this study. The patients underwent DES implantation (n = 11) or BMS implantation (n = 11). In all patients the numbers of CECs were determined before and a week after stent implantation using flow cytometry and the obtained data were compared within and between groups by paired and unpaired Student's t-test, respectively. CECs were defined as cells negative for CD45 (FITC) and highly double positive for CD146 (PE) and CD34 (PE-Cy5) expression. RESULTS: There were no significant differences in the baseline levels of CECs between two groups (p = 0.96). Stent implantation led to a significant increase in CECs compared with the preprocedural levels in the BMS group (p = 0.005) whereas there was a significant decrease in CEC numbers in DES group (p < 0.001). One week after stent implantation CECs count in BMS group was significantly higher compared to DES group (p < 0.001). CONCLUSIONS: The results indicate that patients undergoing DES implantation were subjected to less endothelial injury than patients receiving BMS as indicated by CEC enumeration.