Effects Of Exercise Training And Chlorogenic Acid Supplementation On Hepatic Lipid Metabolism In Prediabetes Mice

Background@#Since prediabetes is a risk factor for metabolic syndromes, it is important to promote a healthy lifestyle to prevent prediabetes. This study aimed to determine the effects of green coffee (GC), chlorogenic acid (CGA) intake, and exercise training (EX) on hepatic lipid metabolism in prediabetes male C57BL/6 mice. @*Methods@#Forty-nine mice were randomly divided into two groups feeding with a normal diet (n=7) or a high-fat diet (HFD, n=42) for 12 weeks. Then, HFD mice were further divided into six groups (n=7/group): control (pre-D), GC, CGA, EX, GC+EX, and CGA+EX. After additional 10 weeks under the same diet, plasma, and liver samples were obtained. @*Results@#HFD-induced prediabetes conditions with increases in body weight, glucose, insulin, insulin resistance, and lipid profiles were alleviated in all treatment groups. Acsl3, a candidate gene identified through an in silico approach, was lowered in the pre-D group, while treatments partly restored it. HFD induced adverse alterations of de novo lipogenesis- and β oxidation-associated molecules in the liver. However, GC and CGA supplementation and EX reversed or ameliorated these changes. In most cases, GC or CGA supplementation combined with EX has no synergistic effect and the GC group had similar results to the CGA group. @*Conclusion@#These findings suggest that regular exercise is an effective non-therapeutic approach for prediabetes, and CGA supplementation could be an alternative to partially mimic the beneficial effects of exercise on prediabetes.

Dietary Antioxidant Intake in Relation to Semen Quality Parameters in Infertile Men: a Cross-Sectional Study

The objective of this study was to assess the association between dietary antioxidant intake and semen quality parameters in infertile men. In this cross-sectional study, dietary antioxidant intake was evaluated in 175 infertile Iranian men by a validated dish-based 106-item semi-quantitative food frequency questionnaire. Men were asked to abstain from ejaculation for at least 72 hours before sample collection. Semen parameters were assessed by a sperm counting chamber and Terminal deoxynucleotidyl transferase dUTP nick end labeling assay methods. Linear quantile regression was used to determine the associations between antioxidant nutrient intake and semen quality parameters (including total sperm count, sperm density, total motility, DNA damage and DNA fragmentation). Mean age of study participants was 32.19 ± 2.34 years. Compared with the lowest quartile, men in the highest quartile of dietary β-carotene and vitamin C intake had lower sperm DNA fragmentation index (Ptrend = 0.042 and Ptrend = 0.03, respectively). Also, dietary intake of beta-cryptoxanthin had a positive association with sperm density (Ptrend = 0.02), and dietary lutein was associated with total sperm count (P(trend) = 0.045). Dietary intake of other antioxidants did not significantly correlate with the indicators related to the quantity and quality of sperm (p > 0.05). These data suggest that dietary intake of some of the antioxidants is associated with semen related parameters.

Epigenotoxic effect of dimethyl sulfoxide on buffalo somatic cells and buffalo-bovine interspecies somatic cell nuclear transfer embryos

Objective: In the present study, we investigated the possible epigenotoxic effect of dimethyl sulfoxide [DMSO] on buffalo fibroblast cells and on reconstructed oocytes during buffalo-bovine interspecies somatic cell nuclear transfer [iSCNT] procedure and its effect on rate and quality of blastocyst which derived from these reconstructed oocytes

Materials and Methods: In this experimental study, cell viability of buffalo fibroblasts was assessed after exposure to various concentration [0.5, 1, 2 and 4%] of DMSO using MTS assay. The epigenetic effect of DMSO was also assessed in terms of DNA methylation in treated cells by flowcytometry. Reconstructed oocytes of buffalo-bovine iSCNT exposed for 16 hours after activation to non-toxic concentration of DMSO [0.5%] to investigate the respective level of 5-methylcytosine, cleavage and blastocyst rates and gene expression [pluripotent genes: OCT4, NANOG, SOX2, and trophectodermal genes: CDX2 and TEAD4] of produced blastocysts

Results: Supplementation of culture medium with 4% DMSO had substantial adverse effect on the cell viability after 24 hours. DMSO, at 2% concentration, affected cell viability after 48 hours and increased DNA methylation and mRNA expression of DNMT3A in fibroblast cells. Exposure of reconstructed oocytes to 0.5% DMSO for 16 hours post activation did not have significant effect on DNA methylation, nor on the developmental competency of reconstructed oocyte, however, it decreased the mRNA expression of NANOG in iSCNT blastocysts

Conclusion: Depending on the dose, DMSO might have epigenotoxic effect on buffalo fibroblast cells and reconstructed oocytes and perturb the mRNA expression of NANOG in iSCNT blastocysts

Effect of vitamin D supplementation on intracytoplasmic sperm injection outcomes: a randomized double-blind placebo-controlled trial

Background: Despite numerous studies indicating an imperative role for reproduction, however, the role of Vitamin D supplementation on outcomes of assisted reproductive techniques remains controversial. This clinical trial was per- formed to evaluate the effect of Vitamin D supplementation 6 weeks prior to intracytoplasmic sperm injection [ICSI] on fertility indices

Materials and Methods: The present study was a double-blind clinical trial conducted on infertile women was randomly allocated into two groups: Vitamin D supplementation [42 participants] and placebo [43 participants]. Serum Vitamin D was measured before and six to eight weeks after treatment, on the day of ovum pick up. Results were analyzed using SPSS16 and fertility indices were compared between the two groups

Results: No significant difference was observed between the intervention and control groups regarding the mean number of oocytes retrieved, percentage mature oocyte, fertilization rate and the rate of good quality embryos [all P>0.05]. But, percentages of the individual with suitable endometrium [7-14 mm thickness] were significantly higher in the Vitamin D compared to control group [P=0.011]. The rate of chemical [47.6 vs. 25.5%, P=0.013] and clinical pregnancy rate [38.1 vs. 20.9%, P=0.019] were also significantly higher in the Vitamin D compared to control group

Conclusion: The present study reveals that consuming Vitamin D for 6 weeks prior to ICSI improves quality of endo- metrium, rate of chemical and clinical pregnancy

Hypothesis: a challenge of overexpression Zfp521 in neural tendency of derived dental pulp stem cells

Neurodegenerative diseases have now become a major challenge, especially in aged societies. Most of the traditional strategies used for treatment of these diseases are untargeted and have little efficiency. Developments in stem cell investigations have given much attention to cell therapy as an alternative concept in the regeneration of neural tissues. Dental pulp stem cells [DPSCs] can be readily obtained by noninvasive procedures and have been shown to possess properties similar to well-known mesenchymal stem cells. Furthermore, based on their neural crest origin, DPSCs are considered to have a good potential to differentiate into neural cells. Zfp521 is a transcription factor that regulates expression of many genes, including ones involved in the neural differentiation process. Therefor based on neural crest origin of the cell and high expression of neural progenitor markers, we speculate that sole overexpression of Zfp521 protein can facilitate differentiation of dental stem cells to neural cells and researchers may find these cells suitable for therapeutic treatment of neurodegenerative diseases

Assessment of PGC1 Alpha -FNDC5 axis in granulosa cells of PCOS mouse model

Background: Polycystic ovarian syndrome [PCOS] is a metabolic and endocrine disorder which is characterized by hyperandrogenism, anovulation or oligomenor-rhea and polycystic ovarian morphology. It is believed that modulation in metabo-lism of granulosa cells of PCOS patients may lead to infertility. One of the metabolic modulators is FNDC5 and its cleaved form, irisin. The axis of PGC1 Alpha - FNDC5 pathway is one of the main factors affecting cellular energy balance the purpose of this study was to evaluate this pathway in granulosa cells derived from PCOS mice model in comparison with control group

Methods: In the present study, PCOS mouse model was developed by injection of dehydroepiandrosterone [DHEA] hormone in 20 mice for a period of 20 days. Also, 20 uninjected mice were used as the control. Meanwhile, a vehicle group consisted of mice which received daily subcutaneous sesame oil injection [n=20]. Relative ex-pressions of PGC1á and FNDC5 in granulosa cells were evaluated by RT-qPCR. Analysis of gene expressions was calculated by the Delta Delta CT method and the relative levels of mRNA were normalized to GAPDH transcript levels. Differences in genes expression among three groups were assessed using one-way ANOVA, Tukey's Post Hoc test

Results: Our results showed that expression of FNDC5 was significantly reduced in granulosa cells of DHEA-induced PCOS mice compared with control and vehicle groups [p<0.05], while there was no significant differences in PGC1 Alpha expression among different groups

Conclusion: Down regulation of FNDC5 transcript level may contribute in metabol-ic disturbance of granulosa cells derived from PCOS ovary apart from PGC1 Alpha levels which remained unchanged

Rational development of a polycistronic plasmid with a CpG-free bacterial backbone as a potential tool for direct reprogramming

Objective: Induced pluripotent stem cells are generated from somatic cells by direct reprogramming. These reprogrammed pluripotent cells have different applications in biomedical fields such as regenerative medicine. Although viral vectors are widely used for efficient reprogramming, they have limited applications in the clinic due to the risk for immunogenicity and insertional mutagenesis. Accordingly, we designed and developed a small, non-integrating plasmid named pLENSO/Zeo as a 2A-mediated polycistronic expression vector

Materials and Methods: In this experimental study, we developed a single plasmid which includes a single expression cassette containing open reading frames of human LIN28, NANOG, SOX2 and OCT4 along with an EGFP reporter gene. Each reprogramming factor is separated by an intervening sequence that encodes a 2A self-processing peptide. The reprogramming cassette is located downstream of a CMV promoter. The vector is easily propagated in the E. coli GT115 strain through a CpG-depleted vector backbone. We evaluated the stability of the constructed vector bioinformatically, and its ability to stoichiometric expression of the reprogramming factors using quantitative molecular methods analysis after transient transfection into HEK293 cells

Results: In the present study, we developed a nonviral episomal vector named pLENSO/Zeo. Our results demonstrated the general structural stability of the plasmid DNA. This relatively small vector showed concomitant, high-level expression of the four reprogramming factors with similar titers, which are considered as the critical parameters for efficient and consistent reprogramming

Conclusion: According to our experimental results, this stable extrachromosomal plasmid expresses reliable amounts of four reprogramming factors simultaneously. Consequently, these promising results encouraged us to evaluate the capability of pLENSO/Zeo as a simple and feasible tool for generation of induced pluripotent stem cells from primary cells in the future

Relationship between potential sperm factors involved in oocyte activation and sperm DNA fragmentation with intra-cytoplasmic sperm injection clinical outcomes

Objective: The present study aimed to simultaneously evaluate the association between expression of three potential factors [post-acrosomal sheath WW domain-binding protein [PAWP], phospholipase Czeta [PLCzeta], and truncated form of the kit receptor [TR-KIT]] as candidates of oocyte activation with fertilization rate and early embryonic development

Materials and Methods: In this experimental study, semen samples were collected from 35 intra-cytoplasmic sperm injection [ICSI] candidates and analyzed according to World Health Organization criteria [2010]. Each sample was divided into two parts. The first part was processed for insemination by density-gradient centrifugation [DGC] and the second part was prepared for assessment of sperm morphology [Papanicolaou staining], DNA fragmentation [transferase dUTP nick end labeling [TUNEL]], and three Sperm-borne oocyte-activating factor [s] [SOAFs]-PLCzeta, PAWP, and TR-KIT

Results: Significant positive correlations existed between the percentages of PLCzeta, PAWP, and TR-KIT with fertilization rate. In addition, significant negative correlations existed between the percentage of DNA fragmentation with the percentages of PLCzeta and PAWP. We did not find a relationship between percentages of PLCzeta, PAWP, and TR-KIT with embryo quality and pregnancy rate [P>0.05]. There was a significant negative correlation between percentage of DNA fragmentation with fertilization and embryo quality

Conclusion: Oocyte activation was associated with the studied sperm factors [PAWP, PLCzeta, and TR-KIT]. These factors might hold the potential to be considered as diagnostic factors in the assessment of semen samples to evaluate their potential to induce oocyte activation. In addition, we observed a significant association between DNA fragmentation with fertilization, as well as embryo quality and expression of PAWP and PLCzeta, which indicated that men with high degrees of DNA fragmentation might require artificial oocyte activation. Whether such action should take place, and its cost and benefits should be evaluated in the future

Chances to have a boy after gender selection by pre-implantation genetic screening are reduced in couples with only girls and without a boy sired by the male partner

Background: Gender selection and family planning have their roots in human history. Despite great interest in these fields, very few scientific propositions exist which could explain why some family do not attain the desired sex. Therefore, the aim of this study was to evaluate whether sex of previous child or children could affect the outcomes of pre-implantation genetic screening [PGS]

Materials and Methods: This historical cohort study including 218 PGS cases referring to Isfahan Fertility and Infertility Center [IFIC]. Couples were grouped as those who their male child passed away or her husbands' has a son[s] from their previous marriage [n=70] and couples who just have daughter [n=148]. Male normal blastocysts were transferred for both groups. The outcomes of PGS including pregnancy, implantation and abortion rates, along with possible confounding factors were compared between the two groups

Results: Significant differences in pregnancy, implantation and abortion rates were observed between couples whose their male partner had/has one boy [n=70] compared to those who have just girl[s] [n=148] despite similar number and quality of male normal blastocyst transferred in the two groups. Confounding factors were also considered

Conclusion: The Y- bearing spermatozoa in male partners with no history of previous boy have lower ability to support a normal development to term, compared to male partners with previous history of boy requesting family balancing

Evidence of oocyte polarity in bovine; implications for intracytoplasmic sperm injection and somatic cell nuclear transfer

Objective: We recently demonstrated spatial regionalization of maternal transcripts and proteins within unfertilized ovine oocyte. Here, we investigated the likelihood of oocyte polarity for the first time in bovine

Materials and Methods: In this experimental study, in vitro matured bovine oocytes were used for manual bisection [into oocyte halve that were near-to [HNS] and far-from [FS] spindle] or trisection [into MII-spindle [S], the spindle-side half [NS], and the distal half unassociated with the spindle [FS]]. Prepared pools of oocyte substructures were used for comparative quantitative real-time polymerase chain reaction [RT-qPCR]. To map the possible preferential sperm entry point [SEP], the spatial relationship between SEP and MII-spindle was measured 5 hours post-fertilization

Results: The proportional amount of maternal mRNA in S oocyte fragment was estimated to be 6 to 11-fold higher than NS and FS counterparts. The relative abundances of Nanog, Oct4, Fgf4 and Tead4 were significantly higher in HNS oocyte fragment compared t0 FS. The relative abundances of Ctnb, Carm1, Rex1, Sox2 and Cdx2 were comparable between HNS and NS oocyte fragments. FS oocyte fragment possessed significantly higher transcripts of Gata4 compared to HNS. The distribution of certain transcripts related to pluripotency and lineage commitment were different depending upon the region of the oocyte; either enriched at S [Tead4, Nanog, Ctnb and Sox2], NS [Oct4], or FS [Gata6]. The SEP in almost [90%] fertilized oocytes was located in MII-hemisphere

Conclusion: The observation of spatial restriction of mRNAs and SEP within MII-oocyte may indicate that the principal forces of oocyte polarity are evolutionary conserved. This may in turn highlight the need for refinements in the methodology of intracytoplasmic sperm injection [where a sperm is injected far from the MII-spindle] and somatic cell nuclear transfer [where a major amount of regulative mRNAs that are associated with MIIspindle is removed during enucleation]

Methylation status of H19/IGF2 differentially methylated region in in vitro human blastocysts donated by healthy couples

Backgrund: Imprinted genes are a unique subset of few genes, which have been differentially methylated region [DMR] in a parental origin-dependent manner during gametogenesis, and these genes are highly protected during pre-implantation epigenetic reprogramming. Several studies have shown that the particular vulnerability of imprinting genes during suboptimal pre- and peri-conception micro-environments often is occurred by assisted reproduction techniques [ART]. This study investigated the methylation status of H19/IGF2 DMR at high-quality expanding/expanded human blastocysts donated by healthy individuals to evaluate the risks linked to ART

Method: Methylation levels of H19/IGF2 DMR were analyzed by bisulfite conversion and sequencing at 18 CpG sites [CpGs] located in this region

Result: The overall percentage of methylated CpGs and the proportion of hyper-methylated clones of H19/IGF2 DMR in analyzed blastocysts were 37.85 +/- 4.87% and 43.75 +/- 5.1%, respectively. For validation of our technique, the corresponding methylation levels of peripheral human lymphocytes were defined [49.52 +/- 1.86% and 50%, respectively]

Conclusion: Considering the absence of in vivoproduced human embryos, it is not possible to conclude that the methylation found in H19/IGF2 DMR is actually normal or abnormal. Regarding the possible risks associated with ART, the procedures should be optimized in order to at least reduce some of the epigenetic risks

Comparative stepwise pattern of reactive oxygen species production during in vitro development of fertilized and nuclear transferred goat embryos

Background: A unique feature of embryo metabolism is production of reactive oxygen species [ROS]. It is well established that during in vitro culture, ROS levels increase over normal ranges observed for embryos developed in vivo. This study evaluates and compares the stepwise pattern of ROS production during in vitro development of reconstructed goat embryos produced by zona-free method of somatic cell nuclear transfer [SCNT]. Furthermore, the pattern of ROS production of SCNT embryos were compared with zona free embryos derived from in vitro fertilization [IVF]

Materials and Methods: In this experimental study, zona-free oocytes, SCNT and IVF embryos at different stages of in vitro development [2, 4, 8, 16-cells, morula, and blastocyst] were used for assessment of ROS production using 2, 7-dichloro dihydroflourescein diacetate [DCHFDA] probe and the result were presented as fold increase or decrease relative zona free oocytes

Results: The relative level of ROS compared to metaphase-II [MII] oocytes insignificantly decrease during early stages post embryo reconstitution and regained its value by 8-cell and morula stage and, significantly increase compared to MII oocytes by blastocyst stage

Conclusion: The pattern of ROS change in SCNT embryos is similar to zona free IVF derived embryos, except it decrease from two cell stage and regain its value at morula stage. The sudden rise in ROS at blastocyst stage, further emphasizes the special need of IVF and SCNT derived embryos during this stage of development

Zinc finger protein 521 overexpression increased transcript levels of Fndc5 in mouse embryonic stem cells.

Zinc finger protein 521 is highly expressed in brain, neural stem cells and early progenitors of the human hematopoietic cells. Zfp521 triggers the cascade of neurogenesis inmouse embryonic stemcells through inducing expression of the early neuroectodermal genes Sox1, Sox3 and Pax6. Fndc5, a precursor of Irisin has inducing effects on the expression level of brain derived neurotrophic factor in hippocampus. Therefore, it is most likely that Fndc5 may play an important role in neural differentiation. To exhibit whether the expression of this protein is under regulation with Zfp521, we overexpressed Zfp521 in a stable transformants of mESCs expressing EGFP under control of Fndc5 promoter. Increased expression of Zfp521 enhanced transcription levels of both EGFP and endogenous Fndc5. This result was confirmed by overexpression the aforementioned vectors in HEK cells and indicated that Zfp521 functions upstream of Fndc5 expression. It is most likely that Zfp521 may act through the binding to its response element on Fndc5 core promoter. Therefore it is concluding that an enhanced expression of Fndc5 in neural progenitor cells is stimulated by Zfp521 overexpression in these cells.

Altered expression of KLC3 may affect semen parameters

Background: KLC3 protein as a member of the kinesin light-chain protein family plays an important role in spermatogenesis, during formation of mitochondrial sheath in the mid piece of the sperm tail

Objective:This study for the first time aims to compare the expression of the KLC3 gene between fertile and infertile individuals

Materials and Methods:Semen samples were collected from 19 fertile individuals who were selected from embryo-donor volunteers and 57 infertile individuals who had abnormal sperm parameters according to world health organization criteria. Sperm parameters using computer assisted sperm analysis and the quantitative KLC3-gene expression using the real-time PCR method were measured

Results: Our results revealed a significant correlations between sperm concentration with relative expression of KLC3 only in infertile groups [r=0.45, p=0.00]. A significant correlation was not found between KLC3 expression and sperm motility; however, the relative expression of KLC3 was significantly higher in asthenozoospermic compared to non-asthenozoospermic individuals

Conclusion: Low expression of KLC3 may result in improper function of midpiece, which has important function in sperm motility. The results of this study show that aberrant expression of KLC3 might be associated with phenomena like oligozoospermia and asthenozoospermia. This article is extracted from student's thesis

Stage-specific profiling of transforming growth factor-beta, fibroblast growth factor and wingless-int signaling pathways during early embryo development in the goat

Objective: This research intends to unravel the temporal expression profiles of genes involved in three developmentally important signaling pathways [transforming growth factor-beta [TGF-beta], fibroblast growth factor [FGF] and wingless/int [WNT]] during pre- and peri-implantation goat embryo development

Materials and Methods: In this experimental study, we examined the transcripts that encoded the ligand, receptor, intracellular signal transducer and modifier, and the downstream effector, for each signaling pathway. In vitro mature MII oocytes and embryos at three distinctive stages [8-16 cell stage, day-7 [D7] blastocysts and day-14 [D14] blastocysts] were separately prepared in triplicate for comparative real-time reverse transcriptase polymerase chain reaction [RT-PCR] using the selected gene sets

Results: Most components of the three signaling pathways were present at more or less stable levels throughout the assessed oocyte and embryo developmental stages. The transcripts for TGF-beta, FGF and WNT signaling pathways were all induced in unfertilized MII-oocytes. However, developing embryos showed gradual patterns of decrease in the activities of TGF-beta, FGF and WNT components with renewal thereafter

Conclusion: The results suggested that TGF-beta, FGF and WNT are maternally active signaling pathways required during earlier, rather than later, stages of pre- and peri-implantation goat embryo development

Overexpression of annexin A1 suppresses pro-inflammatory factors in PC12 cells induced by 1-methyl-4-phenylpyridinium

Objective: Annexin A1 [ANXA1] is suggested to have anti-inflammatory function. However, the precise function of ANXA1 has remained unclear. In this study, we therefore examined the potency of ANXA1 in regulating reactive oxygen species [ROS] production and suppressing pro-inflammatory responses in PC12 cells induced by 1-methyl-4-phenylpyridinium [MPP+]

Materials and Methods: In this experimental study, cDNA of ANXA1 was cloned and inserted to the PGL268 pEpi-FGM18F vector to produce a recombinant PGL/ANXA1 vector for transfection into the PC12 cells. ANXA1 transfected cells were then treated with MPP+. Apoptosis and the content of pro-inflammatory factors including ROS, Interlukin-6 [IL-6], inducible nitric oxide synthase [iNOS] and nuclear factor-kappa B [NF-kappaB] were assessed by flow-cytometry, real-time quantitative polymerase chain reaction [RT-qPCR] and western blot in ANXA1-transfected cells and the data were compared with those obtained from mock and control cells

Results: Data revealed that overexpression of ANXA1 is associated with decreased levels of ROS and expression level of IL-6 and iNOS transcripts, and NF-kappaB protein in MPP+ treated PC12 cells

Conclusion: ANXA1 may be considered as an agent for prevention of neurodegenerative or inflammatory conditions

Peroxisome proliferator-activated receptor gamma activity is required for appropriate cardiomyocyte differentiation

Objective: Peroxisome proliferator-activated receptor gamma [PPAR gamma] is a member of the PPAR nuclear receptor superfamily. Although PPAR gamma acts as a master transcription factor in adipocyte differentiation, it is also associated with a variety of cell functions including carbohydrate and lipid metabolism, glucose homeostasis, cell proliferation and cell differentiation. This study aimed to assess the expression level of PPAR gamma in order to address its role in cardiac cell differentiation of mouse embryonic stem cells [mESCs]

Materials and Methods: In this an intervening study, mESCs were subjected to cardiac differentiation. Total RNA was extracted from the cells and quantitative real time polymerase chain reaction [qPCR] was carried out to estimate level of gene expression. Furthermore, the requirement of PPAR gamma in cardiac differentiation of mESCs, during cardiac progenitor cells [CPCs] formation, was examined by applying the respective agonist and antagonist

Results: The obtained data revealed an elevation in the expression level of PPAR gamma during spontaneous formation of CPCs and cardiomyocytes. Our results indicated that during CPC formation, PPAR gamma inactivation via treatment with GW9662 [GW] reduced expression of CPC and cardiac markers

Conclusion: We conclude that PPAR gamma modulation has an effective role on cardiac differentiation of mESCs at the early stage of cardiomyogenesis

Assessment of DPY19L2 deletion in familial and non-familial individuals with globozoospermia and DPY19L2 genotyping

Background: Globozoospermia is a rare syndrome with an incidence of less than 0.1% among infertile men. Researchers have recently identified a large deletion, about 200 kbp, encompassing the whole length of DPY19L2 or mutations in SPATA16 and PICK1 genes associated with globozoospermia. The aim of this study was to analyze the DPY19L2 gene deletion using polymerase chain reaction technique for the exons 1, 48, 11 and 22 as well as break point [BP] [a] in globozoospermic men

Materials and Methods: In this experimental study, genome samples were collected from 27 men with globozoospermia [cases] and 36 fertile individuals [controls], and genomic analysis was carried out on each sample

Results: Deletion of DPY19L2 gene accounted for 74% of individuals with globozoospermia. DPY19L2 gene deletion was considered as the molecular pathogenic factor for the onset of globozoospermia in infertile men. By quantitative real-time polymerase chain reaction [qPCR], we genotyped DPY19L2 deletion and identified carriers within the population

Conclusion: This technique may be considered as a method for family counseling and has the potential to be used as a pre-implantation genetic diagnosis, especially in ethnic community with high rate of consanguineous marriages

Zeta sperm selection improves pregnancy rate and alters sex ratio in male factor infertility patients: a double-blind, randomized clinical trial

Background: Selection of sperm for intracytoplasmic sperm injection [ICSI] is usually considered as the ultimate technique to alleviate male-factor infertility. In routine ICSI, selection is based on morphology and viability which does not necessarily preclude the chance injection of DNA-damaged or apoptotic sperm into the oocyte. Sperm with high negative surface electrical charge, named [Zeta potential], are mature and more likely to have intact chromatin. In addition, X-bearing spermatozoa carry more negative charge. Therefore, we aimed to compare the clinical outcomes of Zeta procedure with routine sperm selection in infertile men candidate for ICSI

Materials and Methods: From a total of 203 ICSI cycles studied, 101 cycles were allocated to density gradient centrifugation [DGC]/Zeta group and the remaining 102 were included in the DGC group in this prospective study. Clinical outcomes were com- pared between the two groups. The ratios of Xand Y bearing sperm were assessed by fluorescence in situ hybridization [FISH] and quantitative polymerase chain reaction [qPCR] methods in 17 independent semen samples

Results: In the present double-blind randomized clinical trial, a significant increase in top quality embryos and pregnancy rate were observed in DGC/Zeta group compared to DGC group. Moreover, sex ratio [XY/XX] at birth significantly was lower in the DGC/Zeta group compared to DGC group despite similar ratio of X/Y bearings sper- matozoa following Zeta selection

Conclusion: Zeta method not only improves the percentage of top embryo quality and pregnancy outcome but also alters the sex ratio compared to the conventional DGC method, despite no significant change in the ratio of Xand Ybearing sperm population

Upregulation of CD4[+]T-cell derived MiR-223 in the relapsing phase of multiple sclerosis patients

Objective: MicroRNAs [miRNA] are a class of non-coding RNAs which play key roles in post-transcriptional gene regulation. Previous studies indicate that miRNAs are dysregulated in patients with multiple sclerosis [MS]. Th17 and regulatory T [Treg] cells are two subsets of CD4[+] T-cells which have critical functions in the onset and progression of MS. The current study seeks to distinguish fluctuations in expression of CD4[+] T-cell derived miR-223 during the relapsing-remitting [RR] phase of MS [RR-MS], as well as the expressions of Th17 and Treg cell markers

Materials and Methods: this experimental study used real-time quantitative polymerase chain reaction [qRT-PCR] to evaluate CD4[+] T cell derived miR-223 expression patterns in patients that experienced either of the RR-MS phases [n=40] compared to healthy controls [n=12], along with RNA markers for Th17 and Treg cells. We conducted flow cytometry analyses of forkhead box P3 [FOXP3] and RAR-related orphan receptor ?t [ROR?t] in CD4[+] T-cells. Putative and validated targets of miR-223 were investigated in the miRWalk and miRTarBase databases, respectively

Results: miR-223 significantly upregulated in CD4[+] T-cells during the relapsing phase of RR-MS compared to the remitting phase [P=0.000] and healthy individuals [P=0.036]. Expression of ROR?t, a master transcription factor of Th17, upregulated in the relapsing phase, whereas FOXP3 upregulated in the remitting phase. Additionally, potential targets of miR-223, STAT1, FORKHEAD BOX O [FOXO1] and FOXO3 were predicted by in silico studies

Conclusion: miR-223 may have a potential role in MS progression. Therefore, suppression of miR-223 can be proposed as an appropriate approach to control progression of the relapsing phase of MS