Proteins of Leishmania (Viannia) shawi confer protection associated with Th1 immune response and memory generation

<p>Abstract</p> <p>Background</p> <p><it>Leishmania (Viannia) shawi </it>parasite was first characterized in 1989. Recently the protective effects of soluble leishmanial antigen (SLA) from <it>L. (V.) shawi </it>promastigotes were demonstrated using BALB/c mice, the susceptibility model for this parasite. In order to identify protective fractions, SLA was fractionated by reverse phase HPLC and five antigenic fractions were obtained.</p> <p>Methods</p> <p>F1 fraction was purified from L. (V.) shawi parasite extract by reverse phase HPLC. BALB/c mice were immunized once a week for two consecutive weeks by subcutaneous routes in the rump, using 25 μg of F1. After 1 and 16 weeks of last immunization, groups were challenged in the footpad with L. (V.) shawi promastigotes. After 2 months, those same mice were sacrificed and parasite burden, cellular and humoral immune responses were evaluated.</p> <p>Results</p> <p>The F1 fraction induced a high degree of protection associated with an increase in IFN-γ, a decrease in IL-4, increased cell proliferation and activation of CD8⁺T lymphocytes. Long-term protection was acquired in F1-immunized mice, associated with increased CD4⁺central memory T lymphocytes and activation of both CD4⁺and CD8⁺T cells. In addition, F1-immunized groups showed an increase in IgG2a levels.</p> <p>Conclusions</p> <p>The inductor capability of antigens to generate memory lymphocytes that can proliferate and secrete beneficial cytokines upon infection could be an important factor in the development of vaccine candidates against American Tegumentary Leishmaniasis.</p

The Impact of Magnesium Supplementation on Serum Alkaline Phosphatase, Osteocalcin and Fracture Healing in Women with Bone Fracture

Introduction: Magnesium is an essential mineral in bone formation. This nutrient incorporates in bone metabolism and enhances bone mineralization. This study was designed to assess the effect of magnesium supplementation on alkaline phosphatase , osteocalcin, and also callus formation in women with long bone fracture. Methods: In this double-blind randomized placebo controlled trial, 32 women with long bone fracture, aged 20-45 years old, were randomly divided into the Mg group and control, receiving 250 mg magnesium oxide daily and placebo respectively for 8 weeks. Serum alkaline phosphatase and osteocalcin were measured at the beginning and the end point, and also callus formation was checked at the end of study. P value < 0.05 was considered as the significant level. Results: There was no significant difference between two groups in alkaline phosphatase and osteocalcin levels at the beginning and the end of study. Serum alkaline phosphatase and osteocalcin levels were increased in both groups, but they were not statistically significant. Furthermore, the callus formation, which revealed the fracture healing, was not different between 2 groups. Conclusion: This study concluded that Magnesium supplementation did not change the serum markers of bone formation and fracture healing; however, further studies need to approve this finding

Investigating Risk Factors for Cardiovascular Disease Based on Polycystic Ovary Syndrome phenotypes in the 18-14 year Old High School Girls in Shiraz 2009

Introduction: In patients with polycystic ovary syndrome hyperinsulinaemia, insulin resistance, dyslipidemia and hyperglycemia may represent an increased risk for coronary cardiovascular disease .This study aimed to investigate risk factors for cardiovascular disease based on polycystic ovary syndrome phenotypes in Shiraz. Methods: This Cross-sectional study was performed on 3200 students aged 18-14. Demographic survey, clinical signs of androgen excess (acne, hirsutism, alopecia), Ultrasound were applied in order to find the cyst. Tests included prolactin, dehydroepiandrodion sulfate, and oral glucose tolerance test, fasting blood glucose, blood sugar two hours later, triglycerides, cholesterol, high density lipoprotein. Data were submitted to SPSS software, version 11.5 and then analyzed by chi-square tests. Results: The serum cholesterol mean in four phenotypes had a statistically significant relationship with non-PCOS patients(p<0.05). Mean of serum cholesterol in oligomenorrhea, Hyperandrogenism and polycystic ovary phenotype (195.09±30.28) was higher than the other phenotypes. Mean of serum cholesterol and low density lipoprotein(LDL-C) were significantly higher in patients with Hyperandrogenism and polycystic ovarian phenotype(130.046±26.27) and oligomenorrhea, Hyperandrogenism and polycystic ovary syndrome phenotype(138.58±28.34) compared with non-infected individuals. Serum glucose mean in all phenotype was higher than non-infected after two hours and it showed a significant relation in oligomenorrhea and also polycystic ovarian phenotype(98.03 ± 20.98 versus 87.5±12.97) with non-infected individuals. Conclusion: Biochemical factors that lead to increased risk of cardiovascular diseases is increased in patients with polycystic ovary syndrome. Therefore, it should be attended in prevention program

Isolation and Cultivation of Adult Human Keratinocyte Stem Cells for Regeneration of Epidermal Sheets

Background: Keratinocyte stem cell is one of the adult stem cells that inhabits the skin and contributes to skin function and renewal. Adult stem cells are best defined by their capacity to self-renew, and to maintain tissue function for a long period of time. These findings indicate the importance of these cells for clinical applications including regenerative medicine, tissue engineering and gene therapy. In full-thickness damage or injury including burns, the  cultured epidermal autografts (CEAs) may be placed directly onto muscle or fascia. Methods: A small split thickness skin biopsy (1 12أ—'> 2 cm) was obtained aseptically to isolate stem cells. The biopsy was cut into thin pieces and treated with trypsin at 4° C overnight (cold trypsin method) to obtain a single-cell suspension. The cells were seeded at a density of 3×104 cells/cm2 onto a preformed mitomycine-C treated 3T3 cell as feeder  layer  in DMEM medium supplemented with 10% fetal  bovine serum (FBS) and other special supplements. Clonogenic keratinocytes divided and colonies quickly expanded and pushed away the 3T3 feeder layer cells, which then detached from the culture vessel and eliminated with medium changes. Primary cultures were usually subcultured when the cells were in exponential growth phase .Colonies of keratinocytes were expanded and after 7-10 days fused and formed a coherent stratified epithelium. Confluent  cultured  epithelia were detached enzymatically as coherent sheets from the surface of  the culture flasks and transferred onto petrolatum- impregnated gauze.Histological studies of cultured epithelium were also carried out. Results: In our experience from 1 cm2 of skin sample, 2,5- 4×106 cells were obtained. It resulted in keratinocytes suspensions which consisted at least 90% single cells. Cultured keratinocytes proliferated and after 8-10 days became confluent. The area of cultured epithelium detached from T-25 and T-75 culture flasks was approximately 12-15 cm2 and 35-40 cm2 respectively. Histological studies showed that 10-day old cultured epithelium had 3-4 cell layers consisting of small basal cells and big scquamous cells with large nucleus. Also in the basal layer few melanocytes with melanin pigments in the cells cytoplasm were found.The 20-day old cultured epithelium had 8-10 layers consisting of small and round basal cells, scquamous cells and 2-3 layers of keratinized cells. Conclusion: Culture of keratinocyte stem cells could result in multilayer epithelium that creates a good cosmetic appearance upon transplantation. This could re-generate an epidermis that is resistant to trauma and infections. It can be considered as an appropriate substitution in skin loss conditions.   Keywords: Epidermal Sheets, Skin Adult Stem cells, Keratinocyte