Ceratonia siliqua L improved cryodamage of asthenozoospermic specimens: An experimental study

Background: Sperm freezing is an important procedure in assisted reproductive technology. Freezing results in physical and chemical changes in the sperm. Ceratonia siliqua L (C.siliqua) is a tree that has antioxidant properties. Objective: This study aimed to investigate the effect of different concentrations of C.siliqua in a freezing medium on semen parameters, and some biochemical parameters in asthenozoospermic specimens. Materials and Methods: Forty asthenozoospermic specimens (semen specimens with motility < 32%) were obtained from men aged between 20-40 yr according to the World Health Organization criteria. Each sample was divided into 6 groups: I) fresh, II) control, III) 5, IV) 10, V) 20, and VI) 30 μg/ml C.siliqua extract were added to a freezing medium respectively. Then sperm parameters, malondialdehyde, total antioxidant capacity, reactive oxygen species, and sperm DNA assay were evaluated using related protocols after thawing. Results: Data analysis shows that sperm parameter, and total antioxidant capacity level increased at a concentration of 20 μg/ml of C. siliqua extract compared to the other concentrations of C.siliqua extract after cryopreservation and thawing (p < 0.001). Also, the sperm DNA fragmentation assay, reactive oxygen species, and malondialdehyde levels were significantly reduced by adding 20 μg/ml of C. siliqua extract to the sperm freezing medium compared to the other treated groups after cryopreservation (p < 0.001). Conclusion: C.siliqua extract significantly improved sperm parameters after cryopreservation and thawing in asthenozoospermic specimens, and the greatest impact was observed at the 20 μg/ml C.siliqua L extract concentration (p < 0.001). Key words: Asthenozoospermia, Ceratonia, Cryopreservation, Fertility preservation, Infertility, Male

Seminal plasma total antioxidant capacity and vitamin- C levels in asthenozoospermia: a case- control study

"n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Defective sperm function is now recognized as one of the most important causes of male infertility. Seminal plasma possesses a rich source of different enzymatic and non-enzymatic antioxidants such as vitamin C (ascorbic acid) that protect spermatozoa against oxidative stress as one of the mediators of infertility causing sperm dysfunction and low sperm quality. The aim of this study was investigation of seminal total antioxidant capacity and determination of vitamin C effects on sperm motility. "n"nMethods: We designed a case-control study with a total subject of 62 males. Sperm parameters were analyzed according to World Health Organization guidelines (WHO, 1999). Total antioxidant capacity and vitamin C level of seminal plasma were measured in the 32 normozoospermic as the control group and 32 asthenospermic men as the case group using FRAP (Ferric Reducing of Antioxidants Powers) and RP-HPLC (Reverse Phase High Performance Liquid Chromatography) methods, respectively. "n"nResults: Our results indicated that total antioxidant capacity levels in the seminal plasma of asthenospermic men were significantly lower than healthy men (p=0.002). In addition, we found a positive correlation between reduced total antioxidant capacity levels and low sperm motility. Vitamin C levels of seminal plasma in asthenospermic men were statistically lower than control men (p=0.01)."n"nConclusions: It is suggested that asthenospermia could be related to an antioxidant deficiency or it's reduction

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

Background: 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. Methods: 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. Results: Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium cont-aining 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 isolat-ed from foreskin is more than melanocytes isolated from adult eyelashes. Conclusion: 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

Induction of CD4+CD25+FOXP3+ regulatory T cells by mesenchymal stem cells is associated with modulation of ubiquitination factors and TSDR demethylation

Abstract Background Mesenchymal stem cells (MSCs) are known for their ability to induce the conversion of conventional T cells (Tconvs) into induced regulatory T cells (iTregs) in specific inflammatory contexts. Stable Foxp3 expression plays a major role in the phenotypic and functional stability of iTregs. However, how MSCs induce stable Foxp3 expression remains unknown. Methods We first investigated the role of cell–cell contact and cytokine secretion by bone marrow-derived MSCs (BM-MSCs) on the induction, stability, and suppressive functions of Tregs under various experimental conditions that lead to Foxp3 generation by flow cytometry and ELISA respectively. Second, we studied the effect of MSCs on TRAF6, GRAIL, USP7, STUB1, and UBC13 mRNA expression in CD4+ T cells in correlation with the suppressive function of iTregs by real-time PCR; also, we investigated Foxp3 Treg-specific demethylated region (TSDR) methylation in correlation with Foxp3 stability by the high-resolution melting technique. Third, we studied the effect of ex-vivo-expanded BM-MSCs on the induction of transplant tolerance in a model of fully allogeneic skin transplantation. We further analyzed the cytokine secretion patterns in grafted mice as well as the mRNA expression of ubiquitination genes in CD4+ T cells collected from the spleens of protected mice. Results We found that in-vitro MSC-induced Tregs express high mRNA levels of ubiquitination genes such as TRAF6, GRAIL, and USP7 and low levels of STUB1. Moreover, they have enhanced TSDR demethylation. Infusion of MSCs in a murine model of allogeneic skin transplantation prolonged allograft survival. When CD4+ T cells were harvested from the spleens of grafted mice, we observed that mRNA expression of the Foxp3 gene was elevated. Furthermore, Foxp3 mRNA expression was associated with increased TRAF6, GRAIL, UBC13, and USP7 and decreased STUB1 mRNA levels compared with the levels observed in vitro. Conclusions Our data suggest a possible ubiquitination mechanism by which MSCs convert Tconvs to suppressive and stable iTregs