IL-1alpha C376A transversion variant and risk of idiopathic male infertility in Iranian men: a genetic association study

Background: IL-1alpha produced by Sertoli cells is considered to act as a growth factor for spermatogonia. In this study, we investigated the association of the C376A polymorphism in IL-1alpha with male infertility in men referring to the Kashan IVF Center

Materials and Methods: In this case-control study, 2 ml of blood was collected from 230 fertile and 230 infertile men. After DNA extraction, the C376A variant was genotyped by polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP]. In addition, the molecular effects of the C376A transversion were analysed using bioinformatics tools

Results: A significant association was observed between the homozygous genotype CC with male infertility [odds ratio [OR]=1.97, 95% confidence interval [CI]=1.14-3.41, P=0.016]]. Carriers of C [AC+CC] showed a similar risk for male infertility [OR=1.78, 95% CI=1.06-2.99, P=0.030]. Also, allelic analysis showed that the C allele is associated with male infertility [OR=1.43, 95% CI=1.09-1.88, P=0.011]. In sub-group analysis, we found that the AC genotype is associated with asthenozoospermia [OR=2.38, 95% CI=1.03-5.53, P=0.043]. In addition, carriers of C were at high risk for asthenozoospermia [OR=2.25, 95% CI=1.01-4.10, P=0.047]. Also, C allele was significantly associated with oligozoospermia [OR=1.44, 95% CI=1.01-2.06, P=0.049] and non-obstructive azoospermia [OR=1.67, 95% CI =1.04-2.68, P=0.034]. Finally, in silico analysis showed that the C376A polymorphism could alter splicing especially in the acceptor site

Conclusion: This is the preliminary report on the association of IL-1alpha C376A polymorphism with male infertility in the Kashan population. This association shows that the IL-1alpha gene may be a biomarker for male infertility, and therefore needs additional investigations in future studies to validate this

Learning styles and strategies preferences of Iranian medical students in gross anatomy courses and their correlations with gender / 대한해부학회지

The learning approaches can help anatomy teachers design a suitable curriculum in harmony with their students' learning styles. The research objective is to evaluate gross anatomy learning styles and strategies preferences of Iranian medical students at Kashan University of Medical Sciences (KAUMS). This cross-sectional questionnaire-based study was carried out on 237 Iranian medical students. The students answered questions on approaches to learning anatomy and expressed opinions about learning anatomy in medical curriculum. The data were analyzed to disclose statistically significant differences between male and female students. Iranian male and female students were interested in learning anatomy using notes, plastic models, pictures and diagrams, clinical context, dissection and prosection of cadavers; however, they rarely used cross-sectional images and web-based resources. Both groups of medical students used region and system in learning anatomy. However, there existed some striking differences, particularly in having difficulty in studying anatomy using cadaveric specimens, using books alone, and learning it in small groups. Male students were less interested in learning with cadavers than female counterparts. However, female students were more interested in learning anatomy in small groups. This study suggests that instructors should design gross anatomy curriculum based on limitations of using dissection of cadaver in Iranian universities, emphasis on the applied anatomy, and learning of gross anatomy in small groups.

Evaluation of ebselen supplementation on cryopreservation medium in human semen

An effect of cryopreservation on human sperm is sublethal cryodamage, in which cell viability post-thaw is lost more rapidly at later times than in fresh cells. This study examined whether the addition of an antioxidant to cryopreservation medium could improve the post-thaw parameters and evaluation of sperm chromatin quality of cryopreserved human spermatozoa from men with normal semen parameters. Semen samples [n=35] were collected by masturbation and assessed following WHO standards. Individual samples were classified as two portions. One portion [n=10] was for elucidate the concentration of ebselen.Then the samples [n=25] were divided in to 5groups.The first aliquot remained fresh.The second aliquots was mixed with cryopreservation medium.The third aliquots were mixed with cryopreservation medium containing solvent of ebselen.The forth and fifth aliquots were mixed with cryopreservation medium containing 1.25 and 2.5 microm of ebselen.Samples were frozen and thawed samples were assessed for sperm parameters.Three-way ANOVA Multivariate measures were used to assess. According to this assesment the differences are observed in existent groups in post-thaw count, motility index, vitality staining, and morphology and DNA fragmentation. After freezing the media containing of ebselen, DNA fragmentation is significantly different in comparison with control group. ebselen with 1.25 microm dose was significantly associated with post-thaw DNA fragmentation [p=0.047]. Similarly ebselen with 2.5 microm dose was significantly associated with post-thaw DNA fragmentation [p=0.038]. But other parameters were not altered. These results suggest that the addition of ebselen to cryopreservation medium doesnot improve post-thaw parameters and DNA fragmentation of sperm

Expression of Galectin-3 as a testis inflammatory marker in vasectomised mice

Vasectomy, though in some cases are being confronted with irreversibility, has been accepted as an effective contraceptive method. It is estimated that near 2-6% of vasectomised men ultimately show a tendency to restore their fertility. In some cases, vasectomy has been considered as an irreversible procedure due to many post-vasectomy complications causing this debate. The aim of present study was to investigate the pattern of expression of galectin-3, an inflammatory factor secreted by macrophages and immune cells, following the vasectomy in mice testis tissue. In this experimental study, twenty mature male Balb/c mice, aged two months, were divided into two equal groups: sham and vasectomised groups [n=10]. They were sacrificed four months after vasectomy, while the pattern of galectin-3 expression was investigated using a standard immunohistochemistry technique on testicular tissues. Stereological analyses of testes parameters in vasectomised and sham-operated groups were compared by mixed model analysis. Based on observations, although galectin-3 was not expressed in sham-operated group, it was expressed in 40% of testicular tissues of vasectomised mice, like: seminiferous tubules, interstitial tissues and tunica albugina. Also, our result showed a significant alteration in number of germ and sertoli cells of testicular tissue in vasectomised group in comparison to sham-operated group. In addition, the result of mixed model method confirmed a significant reduction in germ and sertoli cells of vasectomised group. The expression of galectin-3 at different parts of testicular tissue in vasectomised group is higher than sham group. This express illustrates the increase of degenerative changes and inflammation reactions in testicular tissue, leading to chronic complications and infertility, after the vasovasostomy

Expression of galectin-8 on human endometrium: molecular and cellular aspects

Background: the up-regulation of galectin-3, galectin-9, and galectin-15 expression in the luminal and glandular epithelium was reported in preparation of the endometrium for embryo implantation at the midlutheal phase. However, no data was available regarding the expression and the distribution pattern of galectin-8 in the human endometrium during a regular menstrual cycle

Objective: the current study designed to investigate the expression and the distribution pattern of galectin-8, a beta-galactoside-binding lectin in the human endometrium during both proliferative and luteal phases of a regular menstrual cycle

Materials and Methods: endometrial biopsies were obtained from the anterior wall of the uterine cavity of 16 women [proliferative phase: n=4, lutheal phase: n=12]. All female patients with mean age of 37.5 years were fertile [range 25-45]. Each biopsy was divided into three pieces; one piece was fixed in formaldehyde for light microscopy and immunohistochemistry. The second portion fixed in glutaraldehyde for scanning electron microscopy and the third portion was prepared for western blot analysis

Results: data of immunoblotting revealed a molecular weight of 34 KD band with high intensity in the lutheal phase samples. The immunohistochemistry staining demonstrated that galectin-8 expressed at a very low concentration during the proliferative phase, but showed a high expression throughout the lutheal phase. The expression of galectin-8 observed in luminal surface epithelium, glandular epithelium and stroma

Conclusion: the up-regulation of the expression of galectin-8 during lutheal phase may suggest galectin-8 as one of the potential molecular marker of the endometrial receptivity. These data propose that galectin-8 may play an important role during the initial events of human embryo implantation

Low doses of celecoxib stimulate human endometrium growth in a three-dimensional culture model

The endometrium plays a pivotal role in implantation and pregnancy. Cyclooxygenase II [COX-2] has an important function in biological processes such as cell proliferation and inflammation. Celecoxib is a selective inhibitor of COX-2 with numerous pharmacologic functions. The aim of present study is to investigate the effects of celecoxib on the human endometrium in a three-dimensional [3D] culture model. In this experimental study, normal human endometria [n=10] obtained from reproductive age women were cut into 1x1 mm sections. Endometrial explants were placed between two layers of fibrin gel. To create the fibrin gel, we poured a thin layer of fibrinogen solution [3 mg/ml in medium 199 [M199]] into each well of a 24-well culture dish and added thrombin enzyme. Endometrial fragments were placed in the center of each well and covered with a second layer of fibrinogen solution. M199 supplemented with L-glutamine, fetal bovine serum [FBS, 5%] and antibiotics were added to each well. The media in each experimental well contained either1, 10 or 50 micro M of celecoxib. At the end of the study, we calculated endometrial tissue growth changes by scoring methods and determined the percentage of angiogenesis. Data were analyzed by the Kruskal-Wallis method. P<0.05 was considered significant. The growth scores were as follows: control [1.37 +/- 0.16], 1 micro M [1.96 +/- 0.28], 10 micro M [2.01 +/- 0.25], and 50 micro M [1.17 +/- 0.14] celecoxib, all of which were significantly different. The angiogenesis percentages were: 25.56 +/- 6.72% [control], 31.98 +/- 6.18% [1 micro M], 42.67 +/- 7.27% [10 micro M] and 23.44 +/- 4.03% [50 micro M], which were not significantly different from each other. Lower celecoxib concentrations had stimulatory effects on the growth of normal endometrium

Co-expression of galectin-3 and alpha v beta 3 integrin at pinopodes of human endometrium

Pinopodes are suggested as biological markers of uterine receptivity, but their molecular components are unknown. Co-expression of galectin-3 and alpha v beta integrin at human pinopodes has been examined in this study to propose a role for them during adhesion phase of embryo implantation. Biopsies were obtained from early and mid luteal phase endometrium of 12 fertile women with regular menstrual periods [25-35 days] and the mean age of 37 years [range 25-45]. Then, they were examined under light and scanning electron microscopy for detection and dating of pinoposes. Using immunofluorescent staining and immunogold electron microscopy, the expression of galectin-3 and alpha v beta integrin in human endometrium and pinopodes was detected. Further, statistical analysis was performed using immunogold electron microscopy to investigate the expression and subcellular distribution of these, before and during the frame of implantation window. The results demonstrated that pinopodes of luminal epithelia cells exhibited immnoreactivity for both galectin-3 and alpha v beta integrin, which was increased statistically significant [p<0.05] at the time of implantation window. Furthermore, area-related distribution of these proteins was found higher in pinopodes compared to the neighboring apical membrane without pinopode. Temporal and spatial expression of galectin-3 and alpha v beta integrin at pinopodes proposes a role for pinopodes in the adhesion of embryo and the involvement of galecting-3 as a binding partner of integrins in the human utero-fetal complex