novel large-scale deletion of the mitochondrial DNA of spermatozoa of men in North Iran

To investigate the level of correlation between large-scale deletions of the mitochondrial DNA [mtDNA] with defective sperm function. In this analytic study, a total of 25 semen samples of the normozoospermic infertile men from North of Iran were collected from the IVF center in an infertility clinic. The swim-up procedure was performed for the separation of spermatozoa into two groups; [normal motility group and abnormal motility group] by 2.0 ml of Ham's F-10 medium and 1.0 ml of semen. After total DNA extraction, a long-range polymerase chain reaction [PCR] technique was used to determine the mtDNA deletions in human spermatozoa. The products of PCR analysis showed a common 4977 bp deletion and a novel 4866 bp deletion [flanked by a seven-nucleotide direct repeat of 5'-ACCCCCT-3' within the deleted area] from the mtDNA of spermatozoa in both groups. However, the frequency of mtDNA deletions in abnormal motility group was significantly higher than the normal motility group [56, and 24% for 4866 bp-deleted mtDNA and, 52, and 28% for 4977 bp-deleted mtDNA, respectively]. It is suggested that large-scale deletions of the mtDNA is associated with poor sperm motility and may be a causative factor in the decline of fertility in men

[Primary culture of human skin melanocyte and comparison of culture in the presence and absence of phorbol ester]

Primary culture takes place following the cell isolation from tissues. Isolation and culture of melanocytes based on their roll in the protection of body against hazardous sun rays, production of skin, cornea and hair color is really important. This study was done to set isolation, culture and proliferation of melanocytes from children foreskin and adult eyelashes, and also comparison of two types of melanocyte culture medium. Human foreskin and eyelash samples were used for melanocyte isolation and culture. After isolation of epidermis from dermis, epidermis cell suspensions were prepared by enzymatic digestion. The isolated cells were cultured in two melanocyte selective culture media. Immunocytochemistary and reverse transcription-polymerase chain reaction [RT-PCR] assays were used for confirmation of isolated and cultured melanocytes. Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium containing phorbol esters not only morphologically but also physiologically and from the aspect of cell adhesion. In addition, the results showed that isolated melanocyte from adult eyelashes are more dendritic than melanocytes isolated from children foreskin. Conversely, our results indicated that the number of cell passages in melanocyte isolated from foreskin is more than melanocytes isolated from adult eyelashes. Melanocytes cultured in selective medium containing convenient growth factors in absence of phorbol esters show more native physiological and adhesive properties. In addition, melanocyte isolated from younger tissues such as foreskin have better proliferative and sub-culturing properties so we suggest isolation and culture of younger tissues.

Lack of association between TRAF1/C5 rs10818488 polymorphism and rheumatoid arthritis in Iranian population

Rheumatoid arthritis [RA] is a multifactorial disorder related to the inflammatory response system with debilitating and painful conditions. Both genetic and environmental factors, with unknown etiology, play important roles in this disease pathogenesis. Recently, TRAF1/C5 [Tumor Necrosis Factor Receptor-Associated Factor 1/Complement Component 5] polymorphism associated with increased risk for RA has been studied in different populations worldwide, and inconsistent results have been obtained, rs 10818488 allele is located on TRAF1/C5 intergenic region, and has been predicted to be functional. A total of 100 sex- and age-matched people including RA patients [n = 50] and healthy individuals [n = 50] from Iran have been entered in this study and genotyped for rs 10818488 [A/G] polymorphism, using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism [PCR-RFLP]. In our study, rs10818488 allele was not associated with risk for RA in Iranian population [p > 0.05, OR = 1.27, 95% CI = 0.72-2.23]. Results revealed that this allele might be population-specific and not to be associated with their corresponding gene pool. However, further analyses are required to clarify other RA-associated markers in our community

Anti tumor activity of salvia officinalis is due to its anti-angiogenic, anti-migratory and anti-proliferative effects

The significant factor contributing to the distant invasion of cancer cells is the ability of tumors to produce large numbers of new blood vessels, known as angiogenesis. Many natural products inhibit angiogenesis. Herein, ethanol extract of Salvia officinalis [SO] has been analyzed for its anti-angiogenic, anti-proliferation and anti-migration activities. The anti-angiogenic effect of the SO extract was evaluated on chicken chorioallantoic membrane [CAM] neovascularisation model, microscopically. The inhibitory effect of the extract on human umbilical vein endothelial cells [HUVECs] migration was tested on the wound-healing model with an inverted microscope. In addition, SO extract was screened for its possible anti-proliferative effects by separately counting HUVECs, Wehi and K562 cells with cell counter against their control wells, So extract exhibited a significant inhibitory activity in CAM assay in a dose dependent manner. CAM angiogenesis was gradually prevented to from at 100 micro g/ml of SO extract, but completely inhibited to form at 200 micro g/ml. After human umbilical vein endothelial cells [HUVECs] were suppressed by dose-dependent SO extract, their migrations were detected by wound healing model, yet they were unable to show a dose response effect on proliferation of the different cells [50-200 micro g/ml]. As observing in this study, SOextract could inhibit proliferation of the different cells at the concentrations above 200 micro g/ml without toxic effect on the cells in doses ranged from 0-500 micro g/ml. These findings indicated that SO extract might be a promising candidate for anti-angiogenic treatment