Use of single-enzyme PCR-restriction digestion barcode targeting the internal transcribed spacers [ITs rDNA] to identify dermatophyte species

Dermatophytes are the most common causative agents of superficial mycoses. Species identification of these fungi is important from therapeutic and epidemiological point of wive. Traditional approaches for identification of dermatophytes at the species level, relying on macroscopic and microscopic features of the colonies, usually are time-consuming and unreliable in many circumstances. Recently a broad varieties of rapid and accurate DNA-based techniques were successfuly utilized for species delineation of dermatophytes. The ITS1-5.8S-ITS2 region of rDNA from various reference strains of dermatophyte species were amplified using the universal fungal primers ITS1 and ITS4.The PCR products were digested by a single restriction enzyme, MvaI. The enzyme was evaluated in both in silico and practical PCR-RFLP assay to find the exact differentiating restriction profiles for each species. To validate the standardized PCR-RFLP system, all tested strains were subjected to sequencing and sequence analysis. The obtained RFLP patterns were specific for many species including T. interdigitale, T. rubrum, T. violaceum, M. persicolor, M. audouinii, M. nanum [A. obtusum] and E. floccosum but were similar for some closely related species such as M. canis / M. ferrugineum. Sequencing of the ITS1-5.8S-ITS2 fragment from all type strains affirmed the RFLP findings. It was practically revealed that the ITS-PCR followed by MvaI-RFLP is a useful and reliable schema for identification and differentiation of several pathogenic species and can be used for rapid screening of even closely related species of dermatophytes in clinical and epidemiological settings

[Inductive effect of deprenyl and dimethyl sulfoxide on proliferation and survival of the mesenchymal stem cells]

Research have been focused on the applying the chemical inducer for transdifferentiation the adult BMSCs into neural cell. So that, at the first should investigate the toxcity effect of the chemical inducer on the induced cells. Plasticity and easy accessibility of bone marrow mesenchymal stem cells is a unique charactristic for treatment of neural disorderies. This study was desgined to determine the inductive effect of Deprenyl and Dimethyl sulfoxide on proliferation and survival of the mesenchymal stem cells. In this experimental study, BMSCs isolated from the adult rat bone marrow and cultured in alpha MEM containing 10% FBS. Cell identity for surface antigens was performed in third passage by immunocytochemistry and multipotancy capacity of BMSCs was done by BMSC differentiation into adipocytes and osteocytes. The cells were exposed to chemical agents [a: the alpha MEM medium supplemented with 2% DMSO, b: the alpha MEM medium supplemented with 10[-8]M Deprenyl] for 24 hours and then transferred to alpha MEM containing 10% FBS cell survival and proliferation was evaluated after the 24, 48, 72 and 96 houres by MTT [3-[4-5-Dimethylthiazolyl-2-y1]-2,5-diphenyltetrazolium romid] test. Data were analyzed using SPSS-16, One-Way ANOVA and Tukey tests. In addition to expression the surface antigens and adipogenic and osteogenic differentiation by BMSCs, MTT test results showed that proliferation and survival of induced-deprenyl and DMSO cells within 48, 72 and 96 hours after the induction was increased significantly than negative control group. Deprenyl increases survival and cell proliferation compared to Dimethyl Sulfoxide. It can be used as cell inducer

[ effects of using retinoic acid during pregnancy on spermatogenesis and steroidogenesis in mature male rat]

The aim of this study was to investigate the effects of using Retinoic Acid during pregnancy on the spermatogenesis, testosterone and gonadotropin hormones in mature male rat. Pregnant rats were injected by 10, 20 and 30 mg/kg Retinoic acid [all-trans Retinoic Acid] intraperitoneally on the 8.5, 10.5, 12.5 and 14.5 days after copulation. An equivalent amount of the vehicle was similarly injected to corresponding control rats. Then animals did child birth. On time of maturity [10 weeks], from male animals in all of groups blood to determined the amount of hormones and the reproductive organs were separated. Morphometerical and Histological changes were studied in epididym and testis among experimental and control groups. The results were evaluated by using one way ANOVA and Tukey- test. Results indicateed that there were significant decrease in LH and FSH hormones, seminiferous tubules diameter, number of spermatocyte, spermatid and leydig cells experimental groups compared with the control group especially in animals which received 30 mg/kg of retinoic acid [p<0.05]. Also RA is the cause of decrease of the epididymis and deferent weight, and sperms in the epididymis, significantly [p<0.05]. Using of Retinoic Acid during pregnancy decreases the normal spermatogenesis and Steroidogenesis in mature male rats

Study on fungal flora in the midgut of the larva and adult of the different populations of the malaria vector anopheles stephensi

Many microorganisms in midgut of mosquito challenge with their host and also other pathogens present in midgut. The aim of this study was presence of non-pathogens microorganisms like fungal flora which may be crucial on interaction between vectors and pathogens. Different populations of Anopheles stephensi were reared in insectary and objected to determine fungal flora in their midguts. The midgut paunch of mosquito adults and larvae as well as breading water and larval food samples transferred on Subaru-dextrose agar, in order to detect the environment fungus. Although four fungi, Aspergillus, Rhizopus, Geotrichum and Sacharomyces were found in the food and water, but only Aspiragilus observed in the midgut of larvae. No fungus was found in the midgut of adults. This is the first report on fungal flora in the midgut of the adults and larvae of An. stephensi and possible stadial transmission of fungi from immature stages to adults. The midgut environment of adults is not compatible for survivorship of fungi but the larval midgut may contain few fungi as a host or even pathogen