MiR-182-5p, MiR-192-5p, and MiR-493-5p Constitute a Regulatory Network with CRISP3 in Seminal Plasma Fluid of Teratozoospermia Patients.

Numerous evidences suggested that microRNAs (miRs) could play an active and significant role during spermatogenesis. Cysteine-rich secretory protein (CRISP3) has a role in inflammatory response and is extremely over-expressed in adolescents with varicocele seminal plasma and modified semen analysis. Nowadays, the miRs expression's association with their target genes is well recognized. The aim of this study was evaluating the association of CRISP3 and four candidate miRs among teratozoospermia (TZ) infertile men. First, we have selected four miRs, miR-182-5p, miR-192-5p, miR-204-5p, and miR-493-5p bioinformatically. After that, RNA was extracted from semen samples of 21 TZ patients and 20 normozoospermia (Norm). Then, their expression levels were assessed using real-time polymerase chain reaction method. In the next step, we quantified the expression of two CRISP3 protein isoforms, targeted by these miRs, using western blotting. According to our results, up-regulation of miR-182-5p, miR-192-5p, and miR-493-5p was observed. MiR-182-5p, miR-192-5p, and miR-493-5p showed good AUC values which can be introduced as possible biomarkers of TZ. In addition, the expression level of the CRISP3 glycosylated (31 kDa) isoform was significantly lower in TZ patients than Norm ones. Notably, in TZ patients, there was a possibly positive correlation of glycosylated CRISP3 expression with normal sperm morphology. According to our results, CRISP3 protein can play a significant role in male infertility especially in maturation formation of spermatozoa. Also, deregulation of the studied miRs, miR-182-5p, miR-92-5p, and miR-493-5p, can suggest a regulatory network between these miRs and CRISP3 isoforms and suggest their regulatory roles in male infertility.

Detailed Investigation of Downstream TLR Signaling in the Follicular Cells of Women with Endometriosis.

BACKGROUND: Inflammatory responses within the peritoneal cavity may result in endometrial dysfunction in women with endometriosis. The true causes of this disease remain poorly understood. It is hypothesized that downstream toll-like receptors (TLRs) inflammatory cytokines in response to pathogens may be associated with endometriosis. So, this study was aimed at evaluating the expression of TLRs signaling and endometriosis-associated inflammatory responses. METHODS: Totally, 20 infertile endometriosis patients and 20 normal women undergoing controlled ovarian stimulation were enrolled. The cellular pellet and supernatant were obtained by centrifugation of follicular fluid (FF). Evaluation of TLRs and their signaling pathway gene expression was performed on cellular pellets using quantitative-PCR. The supernatant was used for determination of cytokine protein expression by ELISA. The results are expressed as mean±SEM and a p<0.05 was considered statistically significant. RESULTS: Quantitative-PCR analysis suggested that TLR1, 5, 6, 7, 8, 10, MYD88, NF-ĸB, IL-10 and TGF-ß genes expression significantly increased in patients compared to the control group (p<0.05). TLR3, 9, INF-ß genes expression was significantly lower in endometriosis than control group (p<0.05). There was no significant difference in the expression of TLR2, TLR4, TIRAP, TRIF, TRAM, and IRF3 between two groups. Also, significant increase in the levels of IL-6, IL-8 and MIF protein in FF of endometriosis group was detected in comparison with normal women (p<0.05). CONCLUSION: The expression of TLR downstream signaling in the follicular cells can initiate inflammatory responses and changes in the FF cytokine profile which in turn may induce endometriosis and infertility disorder.

Transplantation of Mouse Induced Pluripotent Stem Cell-Derived Podocytes in a Mouse Model of Membranous Nephropathy Attenuates Proteinuria.

Injury to podocytes is a principle cause of initiation and progression of both immune and non-immune mediated glomerular diseases that result in proteinuria and decreased function of the kidney. Current advances in regenerative medicine shed light on the therapeutic potential of cell-based strategies for treatment of such disorders. Thus, there is hope that generation and transplantation of podocytes from induced pluripotent stem cells (iPSCs), could potentially be used as a curative treatment for glomerulonephritis caused by podocytes injury and loss. Despite several reports on the generation of iPSC-derived podocytes, there are rare reports about successful use of these cells in animal models. In this study, we first generated a model of anti-podocyte antibody-induced heavy proteinuria that resembled human membranous nephropathy and was characterized by the presence of sub-epithelial immune deposits and podocytes loss. Thereafter, we showed that transplantation of functional iPSC-derived podocytes following podocytes depletion results in recruitment of iPSC-derived podocytes within the damaged glomerulus, and leads to attenuation of proteinuria and histological alterations. These results provided evidence that application of iPSCs-derived renal cells could be a possible therapeutic strategy to favorably influence glomerular diseases outcomes.

Impact of hepatitis B virus and hepatitis C virus infection on sperm parameters of infertile men.

BACKGROUND: Viral hepatitis is one of the health problems which have the effects on the health issues. It seems that hepatitis B virus (HBV) and hepatitis C virus (HCV) infection have negative impacts on the semen quality and male infertility rate. OBJECTIVE: In this study, we aimed to evaluate the effects of HBV and HCV on sperm quality among Iranian infertile men referred to Royan Institute Reproductive Biomedicine Research Center between 2003 and 2014. MATERIALS AND METHODS: This retrospective case-control study included 112 HBV positive infertile men and 47 HCV positive infertile men as case group and 112 HBV negative and HCV negative matched infertile men as a control group. All semen analysis and viral parameters assessment was performed in the central laboratory with the same method and instruments. RESULTS: Sperm count among infertile men with HBV and HCV infection was significantly lower than control group [the mean of the total sperm count 100.95░±â–‘118.59, 118.22░±â–‘141.18, 166.27░±â–‘151.25 (p░<░0.001)]. Sperm motility was significantly decreased in HBV and HCV positive men in comparison to the control group [30.97░±â–‘25.88, 31.09░±â–‘28.72, 40.87░±â–‘23.37, respectively (p░<░0.007)]. The percentage of normal sperm morphology was significantly higher in control group in comparison to HBV and HCV infected group [the mean of the normal semen morphology 3.23░±â–‘3.27, 3.70░±â–‘3.83, 4.51░±â–‘3.15 p░<░0.015]. Although there is a significant decline in liquefaction time in the case group but the viscosity, semen volume, and PH of semen samples were similar in the both case and control groups. CONCLUSION: Our results suggest that HBV and HCV infection are associated with poor sperm quality.

Dietary Vitamin E Is More Effective than Omega-3 and Omega-6 Fatty Acid for Improving The Kinematic Characteristics of Rat Sperm.

OBJECTIVE: Although key roles for dietary vitamin E (VITE) and fatty acid (FA) in fertility have been confirmed, limited data are available on the effects of VITE alone, or a constant level of VITE supplemented by dietary omega-6 and omega-3 FAs in combination on male reproduction. Consequently in this paper, the effects of VITE, sunflower oil, fish oil and their combination on rat sperm were investigated. MATERIALS AND METHODS: We divided 50 mature male Wistar rats into 5 groups (n=10) in a experimental completely randomized design for eight weeks: i. Control (CTR): standard diet; ii. Vitamin E diet (VITE): 2 times greater than recommendations; iii. Sunflower oil group (n-6) [gavaged with 0.5 ml/day/rat sunflower oil+VITE diet]; iv. Fish oil group (n-3): [gavaged with 0.5 ml/day/rat fish oil+VITE diet] and v. n-3+n-6 group [gavaged with 0.3 ml fish oil/day/rat+0.2 ml sunflower oil/day/rat+VITE diet]. The sperm parameters were measured by computer assisted semen analyzer (CASA). All data were analyzed with SPSS software. RESULTS: Feed intake decreased in groups which were administered sunflower oil compared with the other groups (P<0.05). The groups which received only VITE or fish oil+VITE had a significantly higher concentration of sperm compared with the n-6+n-3 and CTR group (P<0.05). VITE and n-3 showed significant improved progressive motility compared to the CTR group, whereas the n-6 and n-6+n-3 groups were in the middle (P<0.05). The highest sperm kinematic parameters were observed in the VITE only group. There was no strong correlation between sperm parameters and blood lipid profiles. CONCLUSION: Dietary VITE and fish oil+VITE can improve sperm quality. Our findings can be a focus for improvements in sperm quantity and motility in fertile animals using only dietary VITE.

Macrophage migration inhibitory factor as a potential biomarker of endometriosis.

OBJECTIVE: To evaluate the expression of MIF, CD74, and COX-2 in normal, ectopic, and eutopic endometrium during the menstrual cycle and to assess MIF level in peripheral blood. DESIGN: The expressions of MIF, CD74, and COX-2 in normal, ectopic, and eutopic endometrium were evaluated with the use of real-time polymerase chain reaction. MIF protein in peripheral blood samples was checked with the use of ELISA. SETTING: Reproductive biomedicine research center. PATIENT(S): Sixteen normal women and 20 women with endometriosis. INTERVENTION(S): Ectopic biopsies were obtained with the use of laparoscopic procedure, and eutopic and control biopsies were obtained with the use of Pipelle. Peripheral blood samples were collected before laparoscopy. MAIN OUTCOME MEASURE(S): The expression of MIF, CD74, and COX-2 in normal, ectopic and eutopic endometrium during the menstrual cycle and the expression level of MIF in peripheral blood samples. RESULT(S): Relative mRNA expression of MIF, CD74, and COX-2 were significantly higher in ectopic endometrium than in eutopic and control endometrium. Also, there were significant differences in expression of these genes in normal, ectopic, and eutopic endometrium during the menstrual cycle. Moreover, women with endometriosis had significantly higher circulating levels of MIF compared with control subjects. CONCLUSION(S): Dynamic expression of MIF, CD74, and COX-2 during the menstrual cycle could play an essential role in reproduction, inflammation, and endometrium reconstruction. A higher expression of these genes in ectopic endometrium can be considered as a molecular biomarker for endometriosis development and pathophysiology. Also, a high level of MIF in blood serum can act as a biomarker in the diagnosis of endometriosis.

Intra-renal arterial injection of autologous bone marrow mesenchymal stromal cells ameliorates cisplatin-induced acute kidney injury in a rhesus Macaque mulatta monkey model.

BACKGROUND: Clinically, acute kidney injury (AKI) is a potentially devastating condition for which no specific therapy improves efficacy of the repair process. Bone marrow mesenchymal stromal cells (BM-MSCs) are proven to be beneficial for the renal repair process after AKI in different experimental rodent models, but their efficacy in large animals and humans remains unknown. This study aims to assess the effect of autologous rhesus Macaque mulatta monkey BM-MSC transplantation in cisplatin-induced AKI. METHODS: We chose a model of AKI induced by intravenous administration of 5 mg/kg cisplatin. BM-MSCs were transplanted through intra-arterial injection. The animals were followed for survival, biochemistry analysis and pathology. RESULTS: Transplantation of 5 × 10(6) cells/kg ameliorated renal function during the first week, as shown by significantly lower serum creatinine and urea values and higher urine creatinine and urea clearance without hyponatremia, hyperkalemia, proteinuria and polyuria up to 84 d compared with the vehicle and control groups. The superparamagnetic iron oxide nanoparticle-labeled cells were found in both the glomeruli and tubules. BM-MSCs markedly accelerated Foxp3+ T-regulatory cells in response to cisplatin-induced damage, as revealed by higher numbers of Foxp3+ cells within the tubuli of these monkeys compared with cisplatin-treated monkeys in the control and vehicle groups. CONCLUSIONS: These data demonstrate that BM-MSCs in this unique large-animal model of cisplatin-induced AKI exhibited recovery and protective properties.

Evaluation of conventional semen parameters, intracellular reactive oxygen species, DNA fragmentation and dysfunction of mitochondrial membrane potential after semen preparation techniques: a flow cytometric study.

PURPOSE: The aim of this study was to evaluate the conventional sperm parameters, level of intracellular reactive oxygen species (ROS), DNA fragmentation (DF) and dysfunction of mitochondrial membrane potential (MMP) after semen preparation techniques with flow cytometry. METHODS: Semen samples were obtained from 28 men with normal semen analysis according to WHO (world health organization). Each was divided into three equal parts for processing with routine techniques: conventional swim up (CSW), direct swim up (DSW) and density gradient centrifugation (DGC). The conventional sperm parameters were evaluated with computer-assisted sperm analyzer (CASA) and the level of intracellular ROS, dysfunction of MMP and DF were determined with flow cytometry procedure. RESULTS: Conventional sperm parameters such as motility, progressive motility and normal morphology increased after sperm processing by CSW and DGC compared to DSW. A significant increase in intracellular H2O2 (p < 0.05) was demonstrated in the CSW versus DSW technique, while processed sperm by the DSW procedure showed a significant increase in the percentage of dysfunction of MMP and intracellular O2(•-) (p < 0.05) when compared with CSW and DGC techniques. Additionally, a high mean of DF (p < 0.05) was observed in the DGC technique as compared to CSW. CONCLUSION: Data from flow cytometry study demonstrated that intracellular H2O2 and DF increased after CSW and DGC processing techniques, respectively, whereas the level of intracellular O2(•-) and dysfunction of MMP only increased after the DSW processing technique.

Association of thrombophilia and polycystic ovarian syndrome in women with history of recurrent pregnancy loss.

PURPOSE: To evaluate the prevalence of thrombophilic disorders in polycystic ovarian syndrome (PCOS) women with history of recurrent pregnancy loss (RPL). MATERIALS AND METHODS: This study was carried out in 184 women with history of RPL, of which 92 of them were diagnosed with PCOS and 92 patients were without known PCOS. The prevalence of thrombophilic disorders was compared between the two mentioned groups. RESULTS: According to the findings, 70.7% of PCOS women with history of RPL had thrombophilic disorders. The prevalence of protein C deficiency was significantly higher in PCOS group compared to the non-PCOS group (21.7% vs. 10.9%, p = 0.04). There was a trend toward higher prevalence of protein S deficiency in PCOS group compared to the control group, but the difference did not reach statistical significance (23.9% vs. 13%, p = 0.05). The prevalence of other thrombophilic disorders such as antithrombin III deficiency, homocysteine elevation, antiphospholipid antibody and Factor V Leiden was comparable between groups. CONCLUSION: The prevalence of thrombophilic disorders was more common in PCOS women than the normal group. The protein C deficiency is associated with PCOS in women with history of RPL. There was a trend toward higher prevalence of protein S deficiency in PCOS women, which needs further study.

Generation of human induced pluripotent stem cells from a Bombay individual: moving towards "universal-donor" red blood cells.

Bombay phenotype is one of the rare phenotypes in the ABO blood group system that fails to express ABH antigens on red blood cells. Nonsense or missense mutations in fucosyltransfrase1 (FUT1) and fucosyltransfrase2 (FUT2) genes are known to create this phenotype. This blood group is compatible with all other blood groups as a donor, as it does not express the H antigen on the red blood cells. In this study, we describe the establishment of human induced pluripotent stem cells (iPSCs) from the dermal fibroblasts of a Bombay blood-type individual by the ectopic expression of established transcription factors Klf4, Oct4, Sox2, and c-Myc. Sequence analyses of fibroblasts and iPSCs revealed a nonsense mutation 826C to T (276 Gln to Ter) in the FUT1 gene and a missense mutation 739G to A (247 Gly to Ser) in the FUT2 gene in the Bombay phenotype under study. The established iPSCs resemble human embryonic stem cells in morphology, passaging, surface and pluripotency markers, normal karyotype, gene expression, DNA methylation of critical pluripotency genes, and in-vitro differentiation. The directed differentiation of the iPSCs into hematopoietic lineage cells displayed increased expression of the hematopoietic lineage markers such as CD34, CD133, RUNX1, KDR, alpha-globulin, and gamma-globulin. Such specific stem cells provide an unprecedented opportunity to produce a universal blood group donor, in-vitro, thus enabling cellular replacement therapies, once the safety issue is resolved.

Calciotropic hormones, insulin resistance, and the polycystic ovary syndrome.

OBJECTIVE: To investigate whether abnormalities in serum concentrations of 1,25-dihydroxyvitamin D [1,25(OH)(2)D], 25-hydroxyvitamin D [25(OH)D], parathyroid hormone (PTH), calcium, and phosphorus were associated with risk of polycystic ovary syndrome (PCOS) and obesity. The possible correlations of the calciotropic hormones with insulin resistance were also examined. DESIGN: Case-control study. SETTING: Department of Genetics, Royan Institute. PATIENT(S): Eighty-five women with PCOS and 115 control women were recruited. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Serum levels of glucose, insulin, total calcium, phosphorus, PTH, 25(OH)D, and 1,25(OH)(2)D were measured in all 200 subjects. RESULT(S): The presence of PCOS had age- and body mass index (BMI)-independent positive effects on serum phosphorus, PTH, 25(OH)D, and insulin concentrations as well as on insulin resistance. Furthermore, overweight/obese (BMI > or =25 kg/m(2)) women with PCOS had significantly decreased levels of 1,25(OH)(2)D and glucose compared with normal-weight (BMI <25 kg/m(2)) women with PCOS. In women with PCOS, phosphorus was correlated negatively with insulin and insulin resistance and positively with 1,25(OH)(2) D. In addition, in normal-weight patients, PTH correlated positively with insulin and insulin resistance. CONCLUSION(S): It is possible that elevated levels of phosphorus and PTH in women with PCOS, at least in part, through their effects on insulin levels and insulin resistance, are involved in pathogenesis of the syndrome.

Association of Ureaplasma urealyticum infection with varicocele-related infertility.

BACKGROUND: The role of seminal colonization of Ureaplasma urealyticum in varicocele-related infertility was investigated. METHODOLOGY: Semen samples were obtained from infertile patients with or without varicocele and healthy controls and were subjected to routine semen analysis and PCR. DNA was extracted by Cadieux method and analyzed by PCR protocol with species-specific primers for U. urealyticum (urease gene). RESULTS: U. urealyticum was detected by PCR in 23 of 146 (15.75%) semen specimens from infertile patients and in 3 of 100(3%) healthy men (P<0.001). Infertile patients with varicocele had higher U. urealyticum colonization [17/81(20.98%)] than those without varicocele [6/65(9.23%), P=0.086] or healthy controls [3/100 (3%), P<0.001].The percentage of sperm cells with motility, volume of semen fluid, concentration of sperm cells, and sperm cell with normal morphology were significantly decreased in infertile men (P<0.001). In the group of varicocele patients with PCR positive for U. urealyticum the volume, count and morphology of semen samples were lower than those in the varicocele patients with PCR negative results, but the differences were not significant (P>0.05). CONCLUSION: Although the colonization of U. urealyticum does not affect the semen quality, the high prevalence of this microorganism in varicocele patients may be an additional negative factor affecting varicocele status and worsening reproductive potential.

Urine macrophage migration inhibitory factor in pediatric systemic lupus erythematosus.

We reported a series of ten patients with lupus nephritis (five patients in the relapse phase and five in the remission phase) and measured the macrophage migration inhibitory factor (MIF), an important pro-inflammatory cytokine with probable role in the pathogenesis of many inflammatory diseases, in their urine samples. MIF/creatinine (Cr) ratio directly correlated with disease activity and it does not have any significant difference between inactive disease and normal ones. We found that the urine MIF/Cr ratio not only differentiates active disease from inactive disease and normal ones but also correlates with the activity indices of renal pathology.