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Bassiri F.,Royan Institute for Animal Biotechnology | Bassiri F.,Islamic Azad University at Dāmghān | Tavalaee M.,Royan Institute for Animal Biotechnology | Shiravi A.H.,Islamic Azad University at Dāmghān | And 4 more authors.
Human Reproduction | Year: 2012

background: Intracytoplasmic sperm injection (ICSI) is the primary treatment for male infertility. However for this procedure, with the exception of visual morphological selection, there is no standardization for sperm selection. Recently, the hypo-osmotic swelling test (HOST) has been proposed to potentially select sperm with intact membranes. The aim of this study is to evaluate the ability of this technique to select functional sperm in terms of apoptosis and morphology, as well as nuclear integrity. methods: A total of 20 semen samples were randomly collected from men who attended the Andrology Unit of the Isfahan Fertility and Infertility Center. Semen samples were washed and exposed to hypotonic conditions, before being fixed and simultaneously assessed for membrane integrity as well as abnormal morphology, DNA fragmentation and protamine deficiency by using Papanicolaou, TUNEL and CMA3 staining techniques, respectively. The remaining semen samples were washed with calcium buffer and stained by Annexin V, then exposed to hypotonic conditions before being assessed for early apoptosis along with membrane integrity. results: HOST grade 'd', followed by grade 'c', showed the highest percentages of healthy sperm, whereas sperm of HOST grade 'g' in which anomalies in terms of apoptosis, abnormal head morphology or nuclear immaturity or membrane damage, were most frequently observed in the samples assessed. conclusions: Integration of HOST into the sperm selection procedure may provide a valuable tool for selection of functional sperm required for ICSI. According to this study, insemination of HOST grade 'g' sperm should be avoided during ICSI. © The Author 2012.

Motiei M.,Royan Institute of Biotechnology | Motiei M.,Payame Noor University | Tavalaee M.,Royan Institute of Biotechnology | Rabiei F.,Royan Institute for Biotechnology | And 4 more authors.
Andrologia | Year: 2013

Heat-shock protein A2 (HSPA2) is a testis-specific member of the HSP70 family known to correlate with sperm maturity, function and fertility. The aim of this study was to compare expression of HSPA2 in fertile and infertile individuals using a recently marketed highly purified polyclonal antibody. Thus, after analysing sperm concentration, motility and morphology in semen sample of 49 individuals with male factor infertility and 47 fertile individuals according to World Health Organization guidelines, we evaluated HSPA2 by microscope florescence, flow cytometry, Western blot and RT-PCR. We observed higher percentage of sperm expressing HSPA2 in anterior and equatorial regions in fertile than infertile individuals by microscope fluorescence. Percentage of sperm expressing HSPA2 in two conditions (fixed permeabilised and capacitated) by flow cytometry showed that percentage of HSPA2-positive sperm in fixed permeabilised and also capacitated samples was significantly different between fertile and infertile individuals. Western blot analysis of HSPA2 in semen samples revealed high variation within the fertile and infertile population. The results of RT-PCR revealed higher expression of HSPA2 in the fertile compared to infertile individuals, but this difference was not significant. According to the results, we suggest that HSPA2 expression is heterogeneously expressed on different part of fixed permeabilised sperm and its expression is significantly higher in fertile compared to infertile individuals. The surface expression of this protein significantly increases following capacitation in both fertile and infertile individuals. HSPA2 expression significantly correlates with sperm concentration and morphology. Therefore, aberrant HSPA2 expression may play an important role in capacitation and fertilisation processes. © 2012 Blackwell Verlag GmbH.

Tavalaee M.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Isfahan Fertility and Infertility Center
Andrology | Year: 2016

Sperm-mediated oocyte activation critically depends upon appropriate expression and assembly of sperm-borne oocyte-activating factors (SOAFs) during spermiogenesis. Several factors have been considered as candidate for SOAFs over the recent years. However, little is known about the expression profile of these candidates and their potential contribution to the clinical outcomes of intra-cytoplasmic sperm injection (ICSI), particularly in globozoospermia. This study investigated the expression profile of PLCζ, PAWP, and TR-KIT and clinical outcomes of ICSI in 12 men with total globozoospermia and compared with 12 fertile individuals. Levels of PLCζ, PAWP, and TR-KIT mRNA in the spermatozoa of fertile men were significantly higher than the corresponding values of the globozoospermic subjects. Interestingly, at protein level, expressions of these factors in the cases assessed were low in globozoospermic individuals. Fertilization rates following artificial oocyte activation (AOA) in the majority of globozoospermic couples were higher than the expected 30% cut-off value reported for individuals with failed or low fertilization rate. Clinical outcomes of ICSI-AOA were dependent on inter-individual variation in globozoospermic couples. None of the SOAFs assessed could provide a greater prediction value with respect to fertilization rate in globozoospermic couples which underwent ICSI-AOA. High fertilization (56.06%) and pregnancy (41.7%) rates accomplished in this study following ICSI-AOA indicated that expression profiles of PLCζ, PAWP, and TR-KIT were low in globozoospermic individuals, and ICSI combined with artificial oocyte activation could mimic physiological calcium changes which occur during fertilization. © 2016 American Society of Andrology and European Academy of Andrology

Khazaie Y.,Royan Institute for Biotechnology | Nasr Esfahani M.H.,Royan Institute for Biotechnology | Nasr Esfahani M.H.,Isfahan Fertility and Infertility Center
International Journal of Fertility and Sterility | Year: 2014

MicroRNAs (miRNAs) are small non-coding single stranded RNA molecules that are physiologically produced in eukaryotic cells to regulate or mostly down-regulate genes by pairing with their complementary base-sequence in related mRNA molecules in the cytoplasm. It has been reported that other than its function in many physiological cell processes, dysregulation of miRNAs plays a role in the development of many diseases. In this short review, the association between miRNAs and some male reproductive disorders is surveyed. Male factor Infertility is a devastating problem from which a notable percentage of couples suffer. However, the molecular mechanism of many infertility disorders has not been clearly elucidated. Since miRNAs have an important role in numerous biological cell processes and cellular dysfunctions, it is of interest to review the related literature on the role of miRNAs in the male reproductive organs. Aberrant expression of specific miRNAs is associated with certain male reproductive dysfunctions. For this reason, assessment of expression of such miRNAs may serve as a suitable molecular biomarker for diagnosis of those male infertility disorders. The presence of a single nucleotide polymorphism (SNP) at the miRNAs' binding site in its targeted mRNA has been reported to have an association with idiopathic male infertility. Also, a relation with male infertility has been shown with SNP in the genes of the factors necessary for miRNA biogenesis. Therefore, focusing on the role of miRNAs in male reproductive disorders can further elucidate the molecular mechanisms of male infertility and generate the potential for locating efficient biomarkers and therapeutic agents for these disorders.

Deemeh M.R.,Royan Institute for Biotechnology | Tavalaee M.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Isfahan Fertility and Infertility Center
Reproductive Sciences | Year: 2015

Artificial oocyte activation (AOA) has shown to improve fertility in severe male infertility following intracytoplasmic sperm insemination (ICSI). However, the effect of AOA on the health status of children has not been studied. This pilot historical cohort study aims to evaluate physical and mental health of 79 and 89 children from 275 and 406 couples undergoing ICSI-AOA using ionomycin and conventional ICSI, respectively. The outcomes assessed were clinical pregnancy, abortion, type of delivery, and health of children (major birth defect, mental and behavior status). No significant differences were observed between the ICSI-AOA and the ICSI groups for these parameters, and the rate of major birth defects were not significantly different between the 2 groups. In this study, AOA has not imposed a greater risk on physical and mental health of children born through AOA, but for such a solid conclusion, further trails with higher number of cases are required and conclusions drawn are limited to this study. © The Author(s) 2014.

Razavi S.H.,University of Isfahan | Nasr-Esfahani M.H.,Royan Institute for Stem Cell Biology and Technology | Nasr-Esfahani M.H.,Isfahan Fertility and Infertility Center | Deemeh M.R.,University of Tehran | And 2 more authors.
Andrologia | Year: 2010

Sperm selection parameters based on morphology and motility for ICSI might not be relevant to chromatin integrity. Thus sperm selection based on sperm characteristics has been suggested. Therefore, the aim of this study was to evaluate the efficiency of the zeta and hyaluronic acid (HA) sperm selection procedures with neat semen, for recovering spermatozoa with normal morphology, protamine content and DNA integrity in infertile men. Semen samples from 77 infertile couples were assessed during this study. Semen analysis was carried out according to World Health Organization criteriarotamine content, DNA integrity and sperm morphology were assessed by chromomycin A3, sperm chromatin dispersion and Papanicolaou staining respectively. The results show that both HA and zeta methods were efficient to recover spermatozoa with normal morphology and protamine content. In terms of the latter parameters, there was no superiority between the two procedures. However, in terms of DNA integrity, the zeta method was more efficient compared with the control and HA procedure and no significant difference was observed between HA and the controls. Therefore, the zeta method appears to be a suitable procedure to recover spermatozoa with normal DNA integrity. © 2009 Blackwell Verlag GmbH.

Nasr-Esfahani M.H.,Royan Institute for Biotechnology ACECR | Nasr-Esfahani M.H.,Isfahan Fertility and Infertility Center | Nasr-Esfahani M.H.,Royan Institute for Reproductive Biomedicine | Deemeh M.R.,Royan Institute for Biotechnology ACECR | And 2 more authors.
International Journal of Andrology | Year: 2012

Spermatozoa contribute to approximately half of the genome of future progeny, and therefore, have a profound impact on embryo development post-fertilization. Sperm selection based on viability and normal morphology does not eliminate the chance for DNA damaged spermatozoa to be inseminated and may account for a considerable percentage of failed embryo development post-ICSI (Intra cytoplasmic sperm injection). Therefore, sperm selection based on functional sperm characteristics to preclude insemination of DNA damaged spermatozoa have paved the way for successful ICSI outcomes. In regard to this, different laboratories have introduced novel procedures to replace traditional or orthodox sperm selection methods. This review attempts to provide information on the scientific bases of each procedure, and pinpoint their advantages and disadvantages. In addition to data from our research, a systematic search on the literature, publications and presentations was carried out using such databases as PubMed and ISI-Web. © 2011 The Authors. International Journal of Andrology © 2011 European Academy of Andrology.

Nasr-Esfahani M.H.,Isfahan Fertility and Infertility Center | Nasr-Esfahani M.H.,Royan Institute Isfahan Campus | Deemeh M.R.,Isfahan Fertility and Infertility Center | Deemeh M.R.,Royan Institute Isfahan Campus | Tavalaee M.,Royan Institute Isfahan Campus
Fertility and Sterility | Year: 2010

Objective: To review different methods for artificial oocyte activation and its impact on intracytoplasmic sperm injection (ICSI). Design: Literature review. Setting: University-based and university-affiliated medical centers. Patient(s): None. Intervention(s): None. Main Outcome Measure(s): None. Result(s): The ICSI procedure improves fertilization rates in cases of male factor infertility; however, fertilization failure still occurs in 2% to 3% of ICSI cycles. The main cause of failed fertilization is failure to complete oocyte activation. The investigators do not use a variety of mechanical, electrical, and chemical methods to mimic the calcium rise necessary to activate oocytes after ICSI. Chemical activation is the most commonly used method for artificial oocyte activation, resulting in high fertilization rates. Conclusion(s): Artificial oocyte activation (AOA) may be useful in selected patients who have low fertilization potential. Further studies are required to establish the biosafety of AOA, and clinical tests are needed to evaluate the activation potential of semen samples for proper patient selection. © 2010 American Society for Reproductive Medicine.

Tavalaee M.,Royan Institute for Animal Biotechnology | Deemeh M.R.,Royan Institute for Animal Biotechnology | Arbabian M.,Royan Institute for Animal Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Animal Biotechnology | And 2 more authors.
Journal of Assisted Reproduction and Genetics | Year: 2012

Purpose: Although, at present, the selection of sperm prior to ICSI is based on motility and morphology, undetectable anomalies, and more importantly damaged DNA are overlooked. In this regard, novel sperm selection procedures have gained much interest. For instance, sperm has been selected by Magnetic-Activated Cell Sorting (MACS) based on early apoptotic marker, the externalization of phosphatidylserine (EPS). Review of the literature has revealed that the efficiency of this technique has been mainly evaluated post Density Gradient Centrifugation (DGC). Therefore, there is a need to prove the efficiency of this technique independent of DGC. In addition, considering the fact that DGC induces EPS due to capacitation and acrosome reaction, therefore, the role of MACS before DGC(MACS-DGC) and MACS after DGC (DGC-MACS) should be assessed. Methods: Semen samples from fifteen infertile men were divided into three separate fractions: control, DGC, and MACS. To carry out DGC-MACS, DGC samples were further divided into two fractions and MACS was carried on the second fractions. Similarly to carry out MACS-DGC, the MACS samples were further divided into two fractions and DGC was carried on the second fractions. Percentages of sperm with normal morphology, DNA fragmentation, protamine deficiency, EPS and caspase-3 activity were determined in each fraction. Results: DGC is more efficient than MACS in separating intact sperm only in terms of normal morphology, DNA and chromatin integrity but not for active caspase. However, a combination of these procedures was more efficient than a single procedure to separate intact sperm for the aforementioned parameters. Comparison of the combined procedures showed only higher efficiency to separate active caspase in the MACS-DGC group. Conclusion: Based on these results, we propose MACS-DGC rather than DGC-MACS to be implemented in clinical settings. © 2011 Springer Science+Business Media, LLC.

Kiani-Esfahani A.,Royan Institute for Animal Biotechnology | Tavalaee M.,Royan Institute for Animal Biotechnology | Deemeh M.R.,Royan Institute for Animal Biotechnology | Deemeh M.R.,Isfahan Fertility and Infertility Center | And 3 more authors.
Systems Biology in Reproductive Medicine | Year: 2012

The aim of this study was to assess the potential of dihydrorhodamine 123 (DHR123) to measure oxidative stress produced by human spermatozoa. The results were compared with 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) that is routinely used for assessment of H2O2 produced by spermatozoa. Fluorescence intensity and percentage R123 and DCF positive sperm were measured by flow cytometry. The optimal condition for assessment of reactive oxygen species (ROS) produced by sperm with DHR123 was 0.05 M for 1 million sperm per ml for 20 minutes. The results of ROS measurement by DHR123 showed a significant correlation (r0.818, P<0.001) with DCFH-DA staining. Immunofluorescence of sperm stained with DHR123 revealed ROS production in the sperm mid-piece. In addition a significant correlation was observed between oxidant production assessed by DHR123 and semen parameters. Therefore, DHR123 may be considered a suitable probe for estimating oxidants produced by human spermatozoa, and can present heterogeneity in oxidant production between different samples. © 2012 Informa Healthcare USA, Inc.

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