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Showing 115 results for Pcr

Hossein Hadinedoushan, Robert Normann,
Volume 3, Issue 2 (7-2005)

Background: Polycystic ovary syndrome (PCOS) is related to obesity and to major metabolic alterations including both insulin resistance and beta-cell dysfunction. Ghrelin was identified as the endogenous ligand for the growth hormone secretagogue (GHS) receptor. The actions of ghrelin are carried out through interaction with specific receptor, named GHS-R. Objective: In a case-control study, we compared the expression of ghrelin and GHS-Rs mRNA by Quantitative Real-time PCR method in studied groups in order to determine the role of ghrelin and GHS-Rs in pathogenesis of PCOS. Materials & Methods: Follicular fluid samples were obtained at oocyte collection from 22 patients undergoing IVF-ET as control and 11 patients were diagnosed as having PCOS. Total RNA was extracted from isolated follicular fluid cells and 2�g RNA was diluted and reverses transcribed using random primers and Superscript II. Specific primers for the ghrelin, GHS-R1a and GHS-R1b were designed. Samples were run in triplicate on an ABI Geneamp 5700 sequence detection system. They were subjected to 40 cycles of amplification under condition 92�C- 20s and 62�C-1min using 3�l diluted cDNA (1:7), 10�l 2X SYBR green, 3�l diluted cDNA. ?-actin mRNA was assayed and then normalized to total RNA measurements for each sample. Results: Age, weight and resulting pregnancies did not vary between PCOS and non-PCOS patients, whereas the BMI and serum testosterone level of PCOS were significantly higher than non-PCOS patients. Quantitative real-time RT-PCR showed that mRNA for ghrelin and GHS-R 1b were detectable in follicular fluid cells from all patients. We failed to find mRNA for GHS-R 1a in any of follicular fluid cells. There were no significant difference in ghrelin and GHS-R1b mRNA expression levels between PCOS and non-PCOS groups. Conclusion: Our findings indicate that ghrelin and ghrelin receptors may not be considered risk factors for pathogenesis of PCOS.
Mohamed Youssry, Batuhan Ozmen, Yasser Orief, Khaled Zohni, Safaa Al-Hasani,
Volume 5, Issue 5 (7-2007)

Fertilization involves direct interaction of the sperm and oocyte, fusion of the cell membranes and :::::union::::: of male and female gamete genomes. The completion of this process and subsequent embryo development depend in part on the inherent integrity of the sperm DNA. Sperm genome quality has been emphasized for several years as playing a major role in early embryogenesis. There is clinical evidence showing that human sperm DNA damage may adversely affect reproductive outcomes and that spermatozoa of infertile men possess substantially more DNA damage than do spermatozoa of fertile men. Testing DNA integrity may help selecting spermatozoa with intact DNA or with the least amount of DNA damage for use in assisted reproductive techniques (ARTs). This review will focus on how sperm DNA is organized, what causes sperm DNA damage and what impact this damage may have on reproductive outcome.
Abdolhossein Rezaeian, Seyed Mehdi Kalantar, Safar Farajnia, Mehrdad Soleimani, Abbas Baghi, Abbas Aflatoonian, Sirous Zeinali,
Volume 5, Issue 5 (7-2007)

Seyed Mohammad Seyedhassani, Massoud Houshmand, Seyed Mehdi Kalantar, Abbas Aflatoonian, Glayol Modabber, Fatemeh Hadipour, Mohammad Hossein Fallahzadeh,
Volume 8, Issue 2 (7-2010)

Background: Mitochondrial transfer RNAs (tRNA) genes are essential components of protein biosynthesis. These genes are hotspots for mutations. These mutations are associated with a wide spectrum of human disease. Many genetic factors are known in assessment of repeated pregnancy loss (RPL). Objective: The aim of this study was analysis of tRNA Thr and tRNA Pro in women with RPL.
Materials and Methods: The nucleotide variations of threonine and proline were investigated in 96 women with idiopathic repeated pregnancy loss. The related mitochondrial area was amplified using a polymerase chain reaction (PCR). The PCR products were demonstrated by 2% agarose gel electrophoresis and all the positive samples were purified and verified by an automated DNA sequencing method.
Results: The sequence analysis revealed 4 mutations in tRNA Thr. These mutations were A15907G in 2 cases (2.08%) A15924G in 3 cases (3.12%) G15928A in 10 cases (10.42%) as the most common mutations and G15930A in 3 cases (3.12%) as a novel mutation. Also the result of tRNApro sequencing showed the T15972C mutation in 1 woman (1.04%) as a novel mutation.
Conclusion: These tRNAs mutations can alter their steady state level and affect the structure of tRNAs. It results in protein synthesis defects and in turn mitochondrial dysfunction. The mutations of these genes may help in the assessment of RPL. Further study of an expanded series of these tRNA mutants is recommended to describe their etiologic role in idiopathic RPL.
Seyed Morteza Seifati, Kazem Parivar, Abbas Aflatoonian, Razieh Dehghani Firouzabadi, Mohammad Hasan Sheikhha,
Volume 10, Issue 1 (7-2012)

Background: Endometriosis is one of the most common gynecologic disorders. It is a complex trait and both genetic and environmental factors have been implicated in its pathogenesis. There is growing evidence indicating that exposure to environmental contaminants is a risk factor for endometriosis. Glutathione-S-Transferase M1 (GSTM1) is one of the genes involved in detoxification of endogenous and exogenous compounds.
Objective: Several studies have indicated an association between GSTM1 null mutation and endometriosis. In this study, the possible association between the GSTM1 gene null genotype and susceptibility to endometriosis in woman from central and southern Iran was investigated.
Materials and Methods: One hundred and one unrelated premenopausal women with endometriosis and 142 unrelated healthy premenopausal women without endometriosis were enrolled in the study. Genomic DNA was extracted from Peripheral blood in all subjects. GSTM1 null genotyping was performed by polymerase chain reaction (PCR).
Results: There was no significant difference between frequencies of GSTM1 null genotype in case and control groups (50.5% Vs. 52.1%, p=0.804). Furthermore, this genotype was not associated with severity of endometriosis in our sample (p=0.77).
Conclusion: further studies involving gene-environment and gene-gene interactions, particularly combination of GSTM1 and other GST gene family polymorphisms are needed.
Maryam Khodadadi, Shiva Basavaiah, Saeid Abediankenari,
Volume 10, Issue 3 (7-2012)

Background: Main function of corpus luteum is progesterone synthesis that is significantly accompanied with an increase in levels of mRNA encoding of steroidogenic enzymes known as luteal markers.
Objective: This study was designed to evaluate effects of lithium chloride on the release of steroid hormones and steroidogenic enzymes in gonadotropin-stimulated rats.
Materials and Methods: Immature 23 days old Wistar rats were divided into 10 groups; each group comprised of 8 rats, and induced with single injection of pregnant mare’s serum gonadotrophin (PMSG) and followed by single injection of human chorionic gonadotropin (hCG). Then, rats were given lithium chloride (LiCl) or saline at 12 hours post-hCG injection. Ovaries were collected in 4-hour interval from 8-24 hour post-hCG injection. Expression pattern of steroidogenic acute regulatory protein (StAR), side-chain cleavage cytochrome P450 (P450scc) and 3β-hydroxysteroid dehydrogenase (3β-HSD) genes were determined by semi-quantitative RT-PCR. In addition, serum levels of progesterone and 17β-estradiol were measured by ELISA.
Results: Our results showed that hCG stimulation of progesterone was markedly diminished and transcript levels of key steroidogenic enzymes were altered in the hormone-stimulated rats following LiCl treatment.
Conclusion: These results suggest that critical steps in the function of corpus luteum are disrupted by lithium. It is concluded that LiCl is an effective factor for suppressing of steroid genes expression.

Zohreh Hojati, Somaye Heidari, Majid Motovali-Bashi,
Volume 10, Issue 4 (8-2012)

Background: About 10% of infertilities with obstructive azoospermia are congenital and caused by CF gene mutations. M469I mutation was observed for the first time in Taiwanese patients. This mutation not only causes CF, but also may be the origin of infertility too.
Objective: In this study, we aimed in designing a rapid, reliable RFLP-PCR procedure for detection of M469I mutation. The correlation and association between M469I mutation with infertility was investigated in this study.
Materials and Methods: one hundred ten patients (90 non obstructive and 20 obstructive) and 60 normal individuals were considered in this study. M469I mutation was detected using RFLP-PCR. This technique was completely designed for M469I genotyping, for the first time in our study. Amplification of the region surrounding the mutation in exon 10 of CFTR gene was then performed. RFLP analysis was carried out using the NdeI restriction enzyme. Results: All genomic DNA samples were genotyped successfully. M469I mutation was observed only in patients group. Therefore, genotype containing mutant allele (GT) has been detected only in the patients group. There was no significant correlation between GT and TT genotypes with infertility (p=0.437).
Conclusion: The M469I mutation has only been observed in Exon 10 CFTR gene of infertile patients, not in the control group. This mutation causes congenital bilateral absence of vaz deferens and finally infertility. This indicates a strong association between the M469I mutation and male infertility. Therefore, this is a CF-causing CFTR mutation that could be considered as a cause of infertility.
Elham Siasi, Ahmad Aleyasin, Javad Mowla, Hamid Sahebkashaf,
Volume 10, Issue 4 (8-2012)

Background: Histones are replaced by protamines to condensate and package DNA into the sperm head during mammalian spermatogenesis. Protamine genes defects have been reported to cause sperm DNA damage and male infertility.
Objective: In this study relationship among some protamines genes family SNPs include PRM1 (C321A), PRM2 (C248T) and TNP2 (T1019C), (G1272C), (G del in 1036 and 1046 bp) were studied in 96 idiopathic infertile men with azoospermia or oligospermia and 100 normal control men.
Materials and Methods: Analysis of SNPs was performed using restriction fragment length polymorphism (PCR-RFLP), single strand conformational polymorphism (PCR-SSCP) and PCR sequencing.
Results: No polymorphisms were found for tested SNPs except for PRM1 (C321A) and TNP2 (G1272C) in which frequency of altered AA and GG genotypes were slightly higher in infertile case group. Statistical analysis showed no significant association related to PRM1 (C321A) p=0.805 and TNP2 (G1272C) loci p=0.654.
Conclusion: These results are consistent with previous studies and indicating that all tested SNPs was not associated with oligospermia and azospermia and idiopatic male infertility in Iranian population.
Arash Davoudi, Alireza Tarang, Seyed Ahmad Aleyasin, Abdolreza Salehi, Ramin Seighalani, Farideh Tahmoressi,
Volume 10, Issue 6 (4-2012)

Background: Fetal DNA in maternal plasma and serum has been shown to be a useful material for prenatal fetal sex determination during early gestational ages. Non-invasive prenatal diagnosis is now possible at 8th week of pregnancy, by maternal blood sample testing.
Objective: The purpose of this study was to evaluate two DNA extraction methods from mother plasma and its routine clinical application in bovine fetus gender determination with non-invasive method.
Materials and Methods: Maternal blood samples were taken from 40 pregnant cows during the 8th-38th weeks of gestation. DNA was extracted from 350 μl of maternal plasma with two salting-out and phenol-chloroform methods. The absorption in A260 and purity (A260/A280) of extracted DNA were detected by ultraviolet spectrophotometer. Three μl of the extracted DNA with phenol-chloroform method was used as a template. The PCR reaction was carried out to amplify the fragments of X and Y chromosomes of amelogenin, TSPY and BC1.2 genes.
Results: The difference between the mean absorption of DNA extracted by phenol-chloroform method and salting-out method was not significant in A260 (p>0.05, p=0.3549), but the difference between mean purity (A260/A280) of DNA extracted by phenol-chloroform method and salting-out method was significant (p<0.001). X chromosome fragment was detected in all 40 samples and Y chromosome fragments were detected in 25 plasma samples which were delivered a male calf. The sensitivity and specificity of test was 100% with no false negative and false positive results.
Conclusion: The results showed that phenol-chloroform method is a simple and sensitive method for isolation of fetal DNA in maternal plasma.
Majid Teremmahi Ardestani, Hossein Hadi Nodushan, Abbas Aflatoonian, Nasrin Ghasemi, Mohammad Hasan Sheikhha,
Volume 11, Issue 1 (4-2013)

Background: Recurrent pregnancy loss (RPL) caused by various genetic and non-genetic factors. After chromosome abnormality, thrombophilia is one of the most important genetic factors that could cause RPL. Factor V Leiden and factor II G20210A mutation were the most common mutations cause thrombophilia in the world.
Objective: The purpose of this study was to determine the frequency of factor V Leiden and prothrombine gene mutations in women with RPL compared with women who had uneventful pregnancies.
Materials and Methods: This case control study evaluates the frequency of factor V-Leiden and factor II G20210 genotypes in 80 women with two or more pregnancy losses, compared with 80 women without adverse pregnancy outcome. The mutations were assessed by PCR-RFLP.
Results: Frequency of the factor V Leiden among cases was 2.5%, which was higher than controls (1.25%), but the difference was not significant. No factor II G20210 mutation was found among cases and controls.
Conclusion: These data did not confirm that factor V Leiden and factor II G20210 mutation might play a role in recurrent pregnancy loss in Iranian women.
Hossein Pashaiefar, Mohammad Hasan Sheikhha, Seyyed Mehdi Kalantar, Tahereh Jahaninejad, Mohammad Ali Zaimy, Nasrin Ghasemi,
Volume 11, Issue 1 (4-2013)

Background: Meiotic genes are very important candidates for genes contributing to female and male infertility. Mammalian MutL homologues have dual roles in DNA mismatch repair (MMR) after replication errors and meiotic reciprocal recombination. The MutL homologs, MLH1 and MLH3 , are crucial for meiotic reciprocal recombination and human fertility. In this study the functional polymorphisms of MLH3 C2531T was investigated in Iranian women with unexplained infertility.
Objective: Investigating the association between a common SNP (single nucleotide polymorphism) C2531T in the MLH3 gene and female infertility.
Materials and Methods: In total, 105 women with unexplained infertility as case group and 100 women with at least one child and no history of infertility or abortion as controls were recruited for this association study. The MLH3 C2531T polymorphism was tested by tetra-amplification refractory mutation system-PCR (4P-ARMS-PCR) method.
Results: The MLH3 2531C and T alleles frequencies were 43.33% and 56.67% among infertile patients, and 61.5% and 38.5% among normal controls, respectively. In the patient and control subjects the CC (Pro 844 Pro) genotype frequency of MLH3 C2531T was 4.76% and 25%, the CT (Pro 844 Leu) genotype was 77.15% and 73%, and the TT (Leu 844 Leu) genotype was 19% and 2%, respectively (p=0.0001).
Conclusion: The presence of the polymorphic allele T leads to an increased risk of 2.09 times (OR=2.09, 95% CI=1.38-3.16; p=0.0001) for developing infertility in relation to the control group. Therefore, our data suggest that the MLH3 C2531T polymorphism can be associated with the risk of unexplained infertility in Iranian women.
Li Hai-Xia, Guo Xin-Yu, Xie Yan, Yuan Qi-Long, Ge Ming-Xiao, Zhang Jin-Yu,
Volume 11, Issue 2 (4-2013)

Background: Aggressive embryo and receptive endometrium are necessary for successful implantation. On this time endometrium transformates to receptive state, which permits embryonic implantation. Studies about embryonic implantation and endometrial receptivity are always a hot spot in the field of reproductive medicine.
Objective: To investigate the expression pattern of Meis1 during peri-implantation in mice endometrium.
Materials and Methods: Mice for experiment were raised in SPF environment. The mice were mated with a female/male ratio of 2:1. The female mice with detected plugs were regarded as pregnant day 1 (pd1). Endometrial tissues were collected respectively on pd1, pd2, pd4, pd5 and pd6. Immunohistochemistry was used to detect the location of Meis1 in mice endometrium. The expression level of mRNA and protein of Meis1 were further detected using Quantitative PCR and Western blotting, respectively.
Results: We found that Meis1 is located in the cytoplasm and membrane of endometrial glandual epithelium cells and the nucleus of endometrial stromal and decidual cells. Both Quantitative RT-PCR and western blotting showed that Meis1 expressed regularly in mice endometrium. Meis1 mRNA expressed weakly on pd1, then significantly increased on pd4 (p=0.018), and achieved to a peak on pd5 (p=0.0012), it showed a decrease trend on pd6. Meis1 protein expressed weakly on pd1 and pd2, then significantly increased on pd4 and pd5 (p=0.0019), it showed a decrease trend on pd6.
Conclusion: Meis1 is dynamically expressed in mice endometrium during peri-implantation. The time that Meis1 expression reaches its peak value is coincident with the implantation window, which implied that Meis1 is closely related with embryonic implantation.
Ramaswamy Suganthi, Vv Vijesh, Sanjay Jayachandran, Jahangir Ali Fathima Benazir,
Volume 11, Issue 3 (5-2013)

Background: Y chromosomal microdeletion is an important genetic disorder, which may arise due to intrachromosomal recombination between homologous sequences in the male specific region of the human Y chromosome. It is frequently associated with the quantitative reduction of sperm. The screening for Y chromosomal microdeletions has a great clinical value.
Objective: To develop a sequence tagged site (STS) based multiplex PCR protocol, which could be specific for the rapid detection of AZF deletions and thereby estimating the frequency of AZF sub deletions in infertile South Indian men.
Materials and Methods: In the current study, PCR based Y chromosomal microdeletion screening analysis was performed in 75 men including 30 non-obstructive azoospermic men, 20 severe oligozoospermic, and 25 normozoospermic fertile men (controls) using 15 known STS primer pairs mapped within the AZF locus. Deletion frequency was estimated after successful PCR amplification.
Results: We designed and optimized a STS based multiplex PCR protocol, which could be helpful for the clinicians to detect the Y chromosomal deletions rapidly and specifically. In our study, we estimated an overall deletion frequency of 36%. Among these 12 (40%) were azoospermic and 6 (30%) were oligozoospermic. No microdeletions were observed in normozoospermic fertile men.
Conclusion: Our Study emphasizes the fact that Y chromosomal microdeletion screening tests are unavoidable in the workup of idiopathic male infertility. Mandatory screening for Y deletions should be done in all azoospermic and severe oligozoospermic patients before undergoing assisted reproductive technology.
Ali Reza Navabazam, Fatemeh Sadeghian Nodoshan, Mohammad Hasan Sheikhha, Sayyed Mohsen Miresmaeili, Mehrdad Soleimani, Farzaneh Fesahat,
Volume 11, Issue 3 (5-2013)

Background: Human dental stem cells have high proliferative potential for self-renewal that is important to the regenerative capacity of the tissue.
Objective: The aim was to isolate human dental pulp stem cells (DPSC), periodontal ligament stem cells (PDLSC) and periapical follicle stem cells (PAFSC) for their potential role in tissue regeneration.
Materials and Methods: In this experimental study, the postnatal stem cells were isolated from dental pulp, preapical follicle and periodontal ligament .The cells were stained for different stem cell markers by immunocytochemistry. To investigate the mesenchymal nature of cells, differentiation potential along osteoblastic and adipogenic lineages and gene expression profile were performed. For proliferation potential assay, Brdu staining and growth curve tests were performed. Finally, all three cell types were compared together regarding their proliferation, differentiation and displaying phenotype.
Results: The isolated cell populations have similar fibroblastic like morphology and expressed all examined cell surface molecule markers. These cells were capable of differentiating into osteocyte with different capability and adipocyte with the same rate. PAFSCs showed more significant proliferation rate than others. Reverse transcriptase PCR (RT-PCR) for nanog, oct4, Alkaline phosphatase (ALP) and glyceraldehydes-3-phosphate dehydrogenease (GADPH) as control gene showed strong positive expression of these genes in all three isolated cell types.
Conclusion: PDLSCs, DPSCs and PAFSCs exist in various tissues of the teeth and can use as a source of mesenchymal stem cells for developing bioengineered organs and also in craniomaxillofacial reconstruction with varying efficiency in differentiation and proliferation.
Mohammad Hasan Sheikhha, Mohammad Ali Zaimy, Saeede Soleimanian, Seyed Mehdi Kalantar, Azam Rasti, Maryam Golzade, Hamid Hoseini Fahraji,
Volume 11, Issue 4 (6-2013)

Background: It has been hypothesized that Y-q microdeletion can account for significant proportion of infertility in men. There are three nonoverlapping regions referred to as the "azoozpermia factors" AZFa, AZFb, and AZFc from proximal to distal part of Y-q. These have been defined as spermatogenesis loci, this region deletions have been shown to be involved in male azoospermic or severe oligoozospermic infertility.
Objective: Evaluation the rate of Y-chromosome microdeletions in infertile men.
Materials and Methods: In this case-control study, 25 azoospermic infertile men candidate for intracytoplasmic sperm injection (ICSI) were selected as case group. For control group, 25 normoozoospemric men were selected. All cases and controls had normal 46XY karyotype. DNA extraction and molecular analysis were done on blood samples. Multiplex-PCR method was done to identify the presence of microdeletion in AZFa, AZFb or AZFc loci. Eight STS primers that include two controls were selected to determine Y-chromosome microdeletions.
Results: 20% (5/25) of all patients have at least one microdeletion in more than one region of AZF loci. Totally 17 microdeletions was observed, one case had deletions in three AZF regions, and 4 cases had deletions in two AZF regions. The rate of deletions was 42% (7/17) for AZFc, 35% (6/17) for AZFa and 23% (4/17) for AZFb.
Conclusion: The molecular DNA analysis could help us to know the real cause of infertility and can give good information for good decision for example in men whit microdeletions who want to undertake ICSI procedure the deletions will be passed to their son.
Fatemeh Mirzaei, Zohreh Farzad-Mahajeri,
Volume 11, Issue 4 (6-2013)

Background: Intrauterine growth retardation (IUGR) contributes significantly to fetal morbidity and mortality, but its etiology is unknown in most cases.
Objective: The aim of this study was to examine the association between inherited thrombophilia and IUGR.
Materials and Methods: A case-control study was performed in a tertiary referral center (Afzalipour Hospital) over 2-years period (2010-2011). Cases (n=25) were women who had pregnancies complicated by IUDR and control subjects (n=25) were women who had normal growth fetuses. All women were tested for inherited thrombophilia at least 4 weeks after delivery. Main outcome measure was prevalence of maternal thrombophlia. Genotyping for factor V Leiden, prothrombin gene (nucleotide G20210A), and MTHFR (C677T) mutation was performed by PCR technique. Protein C, S and antithrombin III activity were determined with a clotting assay (STA-Staclot, France).
Results: The prevalence of hereditary thrombophilia was 68% (n=17) in IUGR group, and 32% (n=8) in control group (OR: 1.5, p=0.011, 95% CI: 1.3-14.8). The frequency of MTHFR (C677T) gene mutation (p=0.037; OR: 3.69) and protein S deficiency (p=0.034; OR: 5.41) was significantly increased in the group with IUGR compared with the control group. There was no significant difference between the two groups in prothrombin G20210A mutation (p=0.490) and protein C deficiency (p=0.609). A significant difference in the frequency of multiple thrombophilias was detected between the two groups (p=0.009).
Conclusion: This study revealed that protein S deficiency and MTHFR gene mutation are more prevalent in pregnancies with IUGR.
Maryam Afrakhteh, Atossa Mahdavi, Hadi Beyhaghi, Afshin Moradi, Sima Gity, Shirin Zafarghandi, Zahra Zonoubi,
Volume 11, Issue 4 (6-2013)

Background: Sexually transmitted infections (STIs) are among the most common causes of illness in the world and have far-reaching health, economic and social consequences for many countries. Failure to diagnose and treat STIs at an early stage may result in serious complications and sequels.
Objective: This study aimed to determine the prevalence of Chlamydia trachomatis infection in patients who remain symptomatic after completion of their first episode of treatment for STI.
Materials and Methods: We conducted a cross-sectional study on 49 patients suffering from symptoms or signs of sexually transmitted infections despite their first complete anti STI treatment. Conducting physical exam and smear preparation from vaginal discharge, diagnosis was confirmed by Polymerase chain reaction (PCR) method on every patient’s first-voided urine sample.
Results: Among the etiologic factors investigated in this study, Chlamydia was reported in 17 patients. Trichomoniasis, Candidiasis, Gonorrhea and nonspecific germs were next organisms with 11, 9, 6 and 6 patients, respectively. Sixteen specimens were PCR positive (32.65%), while 33 patients had negative PCR results (67.34%) for Chlamydia trachomatis.
Conclusion: Gonorrheal infection was the most prevalent infection in patients with completed treatment (6/10), which must be remembered in patients follow ups, because this prevalence warrants empirical therapy for Gonorrheain similar clinical conditions. Chlamydia trachomatis was the responsible organism in approximately a quarter of patients (17/75) who despite their full compliance on anti-Chlamydial treatment still suffered from signs and symptoms of STI. This rate also recommends empirical therapy for Chlamydia trachomatis in the similar clinical signs and symptoms.
Saeede Soleimanian, Seyyed Mahdi Kalantar, Mohamad Hasan Sheikhha, Mohamad Ali Zaimy, Azam Rasti, Hossein Fazli,
Volume 11, Issue 5 (7-2013)

Background: In human, about 25% of implanted embryos are losing 1-2 week following attachment to the uterus. A subset of this population will have three or more consecutive miscarriages which define as repeated pregnancy loss (RPL). Introducing the assisted reproductive technologies (ARTS) made a chance for infertile couples to solve their childless problem.
Objective:  This study was conducted to evaluate the incidence of Y-chromosome AZF region's micro-deletions in male partners of couples with recurrent miscarriage (RM).
Materials and Methods:  Thirty male partner of couples with RM and thirty infertile males, who referred to the Yazd Research and Clinical Center for Infertility were recruited to this study. In addition, 30 healthy men were screened as a control group from the same center. After DNA extraction using salting out method, the multiplex-PCR was done for amplifying 8 known STSs proximal to the AZF region of the Y-chromosome. The results were compared between the groups using Fisher's exact t-test and p<0.05 was considered statistically significant.
Results:  Of the 30 infertile males, 5 (16.6%) cases were associated with the AZF region micro-deletions of DYF87S, DYF84S1, DYF83S1 and DYF51S1, STSs. But in the fertile and RM male groups was found no deletions similar to those, of the infertile males (p=1.0). Instead 4 (13.3%) cases of the RM group males had different micro-deletions included DYS220 (AZFb, sY129), DYS262, DYF8551, and DYF8651, STSs. The AZFc locus of Y-chromosome micro-deletions have a significant role in RM (p=0.045).
Conclusion:  It seems that the Y-chromosome AZF region's micro-deletions are associated with RM, and we recommend adding this AZF region STSs into infertility analyzing panels.
Mohammad Ali Zaimy, Seyyed Mehdi Kalantar, Mohammad Hasan Sheikhha, Tahere Jahaninejad, Hossein Pashaiefar, Jalal Ghasemzadeh, Mahnaz Zahraei,
Volume 11, Issue 6 (9-2013)

Background: About 15% of couples have infertility problems which 40% of them are related to the male factors. Genetic factors are candidate for about 10% of male infertility conditions. Among these, AZFa, AZFb, AZFc and AZFd regions on the Yq are considered most important for spermatogenesis. Microdeletions of these regions are thought to be involved in some cases of azoospermic or oligospermic infertile men.
Objective: We studied the prevalence of AZF microdeletions among Iranian infertile men with non-obstructive azoospermia and oligospermia.
Materials and Methods: A total of 50 Iranian azoospermic and oligospermic infertile men were selected for case group and 50 men with normal spermogram as control group. The molecular study of Y chromosome microdeletions was done by multiplex polymerase chain reaction (M-PCR) method by using of 13 sequence tagged site (STS) markers from AZF region.
Results: Four (8%) patients showed Y chromosome microdeletions among case group, deletion in AZFc region was the most frequent (80%) followed by AZFb (20%), in AZFa and AZFd region we did not detect any deletions. No deletion was detected in control group; the ratio of Y chromosome microdeletion in azoospermic men was higher than this ratio in oligospermic men [19% (3/16) among azoospermic men and 3% (1/34) among oligospermics]. Serum FSH level in men with microdeletions was higher than this level in men with no deletions (p=0.034).
Conclusion: Because of relatively high prevalence of microdeletions on the long arm of Y chromosome among Iranian azoospermic and oligospermic patients, screening of this microdeletion may be advised to infertile men particularly azoospermic and oligospermic men before using assisted reproductive treatments.
Fatemeh Kheradmand, Issa Nourmohammadi, Mohamad Amin Ahmadi-Faghih, Mohsen Firoozrai, Mohammad Hossein Modarressi,
Volume 11, Issue 6 (9-2013)

Background: The impact of cadmium (Cd) on male infertility may be related to the interaction with metal-binding proteins known as metallothioneins (Mts). Trace elements like zinc (Zn) have protective effects on testicular damage induced by Cd.
Objective: We determined the effect of Zn and low-dose Cd pre-treatment on the expression of Mt1 and Mt2 genes on testicular Sertoli cells.
Materials and Methods: The cultured TM4 mouse sertoli cells were treated with 50 μM ZnSO4 (Zn pre-treated group; ZnPG), 2 μM CdCl2 (Cd pre-treated group; CdPG), or distilled water (DW pre-treated group; DWPG). After 18 hour, all of these groups were exposed to 100 μM CdCl2 for different periods of time (1, 2, 3, and 6 hours). There was also a control group for all three groups, which was treated only with distilled water (without Cd or Zn pre-treatment). Cellular viability, Zn and Cd concentrations and gene expression were assessed by MTT, atomic absorption spectrometry and real time PCR methods, respectively.
Results: The expression of Mt1 and Mt2 genes in ZnPG, CdPG, and DWPG was greater than the control group (p=0.02 and p=0.01, respectively). Cd concentrations in CdPG and DWPG were greater than the control group (p=0.00). Expression of both genes in ZnPG and CdPG increased after 3 hours of treatment and Cd concentration decreased simultaneously, which was more obvious in ZnPG.
Conclusion: Zn and short term low-dose Cd pre-treatment might reduce the adverse effects of Cd by increasing expression of Mts genes in Sertoli cells. The protective effect of Zn was stronger than Cd.

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