ABSTRACT
Background: Therapeutic potential of in vitro maturation [IVM] in infertility is growing with great promise. Although significant progress is obtained in recent years, existing IVM protocols are far from favorable results. The first aim of this study was to investi-gate whether two step IVM manner change reactive oxygen species [ROS] and total antioxidant capacity [TAC] levels. The second aim was to find the effect of alpha lipoic acid [ALA] supplementation on oocyte maturation rate and on ROS/TAC levels during IVM
Materials and Methods: In this experimental study, mouse germinal vesicle [GV] oocytes divided into cumulus denuded oocytes [DOs] and cumulus oocyte complexes [COCs] groups. GVs were matured in vitro in the presence or absence of ALA only for 18 hours [control] or with pre-culture of forskolin plus cilostamide for an additional 18 hours. Matured oocytes obtained following 18 and 36 hours based on experimental design. In parallel, the ROS and TAC levels were measured at different time [0, 18 and 36 hours] by 2',7'-dichlorodihydrofluorescein [DCFH] probe and ferric reducing/antioxidant power [FRAP] assay, respectively
Results: Maturation rate of COCs was significantly higher than DOs in control group [P<0.05], while there was no significant difference between COCs and DOs when were pre-cultured with forskolin plus cilostamide. ROS and TAC levels was increased and decreased respectively in DOs after 18 hours while in COCs did not change at 18 hours and showed a significant increase and decrease respectively at 36 hours [P<0.05]. ROS and TAC levels in the presence of ALA were significantly decreased and increased respectively after 36 hours [P<0.05] whereas, maturation rates of COCs and DOs were similar to their corresponding control groups
Conclusion: ALA decreased ROS and increased TAC but could not affect maturation rate of both COCs and DOs in one or two step IVM manner
ABSTRACT
Cryopreservation of ovarian tissues and pre-antral follicles is a promising prospect for preservation of women fertility. The aim of this study was to evaluate the in vitro developmental competence of mouse vitrified pre-antral follicles in comparison to isolated pre-antral follicles derived from vitrified ovaries in the presence of alpha lipoic acid [ALA]. Pre-antral follicles derived from fresh, vitrified-warmed ovarian tissues and vitrified-warmed pre-antral follicles were cultured individually with or without ALA, followed by adding hCG to induce ovulation. The follicle growth, oocyte maturation, and embryo development were assessed. The diameter and development of follicles, oocyte maturation and embryo development rates were significantly higher in ALA supplemented groups compared to the respective ALA-free conditions groups. Aforementioned parameters were significantly higher in vitrified-warmed follicles in comparison to follicles derived from vitrified-warmed ovaries. These findings support a superior performance of pre-antral follicles when vitrified rather than when isolated from vitrified ovaries with regard to increasing the rates of developmental parameters. Moreover, ALA improves the in vitro maturation of pre-antral follicles in vitrified and non-vitrified samples.
ABSTRACT
Cryopreservation of pre-antral follicles is a hopeful technique to preserve female fertility. The aim of the present study was to evaluate reactive oxygen species [ROS] and total antioxidant capacity [TAC] levels of mouse vitrified pre-antral follicles in the presence of alpha lipoic acid [ALA]. Isolated pre-antral follicles [140-150 micro m in diameter] were divided into vitrified-warmed and fresh groups. Each group was subjected to in vitro maturation with or without ALA for 12 days, followed by adding human chronic gonadotropin to induce ovulation. In vitro fertilization was performed to evaluate their developmental competence. In parallel, the amount of ROS and TAC were assessed after 0, 24, 48, 72, and 96 h of culture by 2',7'-dichlorofluorescin assay and ferric reducing/antioxidant power assay, respectively. The respective rates of survival, antrum formation, and metaphase II oocytes were significantly higher in ALA-supplemented groups compared to the groups not treated with ALA. TAC and ROS levels were significantly decreased and increased, respectively during the culture period up to 96 h in the absence of ALA in both vitrified and non-vitrified samples. However, with pretreatment of ALA, TAC levels were increased significantly and remained constant up to 96 h in vitrified-warmed pre-antral follicles, while ROS levels completely returned to the level of starting point after 96 h of culture in the presence of ALA. Pretreatment of ALA positively influences development of pre-antral follicles in vitrified and nonvitrified samples through increasing follicular TAC level and decreasing ROS levels
ABSTRACT
Onychomycosis is a nail disorder associated with aesthetic problems, discomfort, physical injury and loss of dexterity. The purpose of the present study was to isolate and identify the causative fungi of Onychomycosis in 2402 patients in Kermanshah Province, western Iran in 1994 to 2010. Mycologic assessment was carried out by standard methods including either microscopic or cultural procedures. Direct microscopy of the nail clips was positive in 1086 [45.2%] and fingernail and toenail onychomycoses were recognized in 773 [71.1%] and 313 [28.8%], respectively. Yeasts were detected in 853 [78.5%], dermatophytes in 201 [18.5%] and non-dermatophyte fungi in 32 [2.9%] patients. The results of fungal culture showed Candida albicans isolated in 384 [45.0%] and other Candida spp. isolated in 361 [54.0%] of the cases as the most common agents of Onychomycosis while among dermatophytes, Trichophyton rubrum was found in 63 [37.0%] of the cases as the main dermatophytic agent followed by T. mentagrophytes 32 [15.9%] and Epidermophyton Flocosum 30 [17.6%]. Among the non-dermatophyte moulds, Aspergillus Flavus was the most prevalent species 12 [37.5%] followed by A. niger 8 [25.0%] and A. Fumigatus 4 [12.5%]. Moreover, 139 [12.8%] samples with positive direct microscopy yielded no growth. The highest rate of Onychomycosis was found in patients between 30-40 years of age. In sum, the current results identified the aetiological agents and primary epidemiological aspects of Onychomycosis in west Iran
Subject(s)
Humans , Female , Male , Incidence , Fungi , Candida , TrichophytonABSTRACT
In spite of extensive efforts to improve in vitro oocyte maturation, the obtained results are not very efficient. This study was conducted to assess impacts of cAMP elevating agents and alpha lipoic acid [ALA] on in vitro oocyte maturation and fertilization. Mouse germinal vesicle [GV] oocytes were categorized into cumulus denuded oocytes [DOs; n=896] and cumulus oocyte complexes [COCs; n=1077] groups. GV oocytes were matured in vitro with or without ALA; [I] without the meiotic inhibitors; [II] supplemented with cilostamide; [III] supplemented with forskolin and [IV] supplemented with Forskolin plus cilostamide. The obtained metaphase II [MII] oocytes were subjected to in vitro fertilization. Independent-samples t-test and ANOVA were used for data analysis. A p-value less than 0.05 was considered to be statistically significant. The COCs maturation, fertilization and two cell embryo rates were higher than those of DOs in the control group, while no significant difference was observed between relevant COCs and DOs when they were cultured with cilostamide meiotic inhibitors in two step manner. Combined treatment of cilostamide and forskolin significantly elevated the developmental rates in both COCs and DOs as compared to other groups. The developmental rates of COCs and DOs in the presence of ALA were similar to their respective groups without ALA. cAMP elevating agents were more effective on DOs than COCs with regard to rates of maturation and fertilization. However, ALA did not affect the developmental rates of both COCs and DOs in in vitro maturation in one or two step manner