Screening of two neighboring CFTR mutations in Iranian infertile men with non-obstructive azoospermia

The genetic association between cystic fibrosis transmembrane conductance regulator [CFTR] gene mutations and male infertility due to congenital bilateral absence of vas deferens [CBAVD] is well established. Mutant CFTR, however may also be involved in the etiology of male infertility in non-CBAVD cases. The present study was conducted to estimate the frequency of DELTA I507 and DELTA F508 CFTR gene mutations in Iranian infertile males. We undertook the first study of association between these CFTR mutations and non-obstructive azoospermia in Iran. In this case-control study, 100 fertile healthy fathers and 100 non-obstructive azoospermia's men were recruited from Isfahan Infertility Center [IIC] and Sari Saint Mary's Infertility Center, between 2008 and 2009. Screening of F508del and I507del mutations was carried out by the multiplex-ARMS-PCR. Significance of differences in mutation frequencies between the patient and control groups was assessed by Fisher's exact test. The DELTA F508 was detected in three patients. However there are no significant association was found between the presence of this mutated allele and infertility [OR=9.2 [allele-based] and 7.2 [individual-based], P=0.179]. None of the samples carried the DELTA I507 mutation. Altogether, we show that neither DELTA I507 nor DELTA F508 is involved in this population of Iranian infertile males with non-obstructive azoospermia

Association between serum paraoxonase 1 activities [PONase/AREase] and L55M polymorphism in risk of female infertility

The risk of developing female infertility has been associated with gene polymorphisms that decrease the activity of enzymes involved in systemic Oxidative Stress [OS]. In this study, PON1 L55M polymorphism for association with susceptibility to infertility was investigated among Iranian female population. Samples from 120 Iranian females [20 endometriosis; 30 Polycystic Ovary Syndrome [PCO]; 70 controls] were analyzed and PCR-RFLP assay was used to determine the PON1 rs854560 [L55M] frequencies. The paraoxonase [PONase] and arilesterase [AREase] activities of PON1 enzyme were also assessed in order to investigate the association between serum PON1 activities, female infertility, and PON1 L55M polymorphism. The women with a MM genotype [p=0.021; OR=2.55] showed more possibilities of experiencing infertility than those with a LM genotype [p=0.039; OR=1.91]. According to LSD test, endometriosis subjects had significantly lower paraoxonase enzyme activity compared to control group [p=0.0024; CI=95%]. No significant difference was found in women with PCOS for both PONase and AREase activity in comparison with control group [p=0.469; CI=95%]. Furthermore, PON1 activities were the highest in LL genotype followed by LM and then MM genotype [MM

Role of XmnIgamma[G] polymorphism in hydroxyurea treatment and fetal hemoglobin level at Isfahanian intermediate beta-thalassemia patients

beta-thalassemia is the most common monogenic disorder in human. The [C[right wards arrow]T] polymorphism at -158 upstream region of the gamma[G]-globin gene and pharmacological factors such as hydroxyurea have been reported to influence gamma-globin gene expression and the severity of clinical symptoms of beta-thalassemia. In the present study, 51 beta- thalassemia intermediate patients were studied. Xmn1gamma[G] polymorphism genotype was determined using Tetra-Primer ARMS-PCR technique. Hemoglobin [Hb] and fetal hemoglobin [HbF] levels were determined by gel electrophoresis. Of 51 patients, 35 [68.6%] patients were heterozygous [CT] and 16 [31.4%] patients were homozygous [CC]. Of 30 patients under treatment by hydroxyurea, 20 [66.7%] patients were heterozygous [CT] and 10 [33.3%] patients were homozygous [CC]. Our results demonstrated that in the heterozygous [CT] genotype, the Hb [9.58 +/- 1.25 gm/dl] and HbF [89.30 +/- 21.87] levels were significantly higher in comparison with homozygous [CC] genotype [7.94 +/- 1.34 gm/dl and 70.32 +/- 40.56, respectively]. Furthermore, we observed that after drug usage, the Hb and HbF levels in patients with heterozygous [CT] genotype [0.7 +/- 1.26 gm/dl and 5.95 +/- 14.8, respectively] raised more in comparison with homozygous [CC] genotype [0.26 +/- 1.43 gm/dl and 0.8 +/- 1.31, respectively]. Hb and HbF levels in the patients carrying T allele are increased significantly, and they also response to hydroxyurea treatment

Multiplex PCR based screening for micro/partial deletions in the AZF region of Y-chromosome in severe oligozoospermic and azoospermic infertile men in Iran

Infertility is a health problem which affects about 10-20% of married couples. Male factor infertility is involved approximately 50% of infertile couples. Most of male infertility is regarding to deletions in the male-specific region of the Y chromosome. In this study, the occurrence of deletions in the AZF region and association between infertility and paternal age were investigated in Iranian men population. To assess the occurrence of Y chromosomal microdeletions and partial deletions of the AZF region, 100 infertile men and 100 controls with normal spermatogenesis were analyzed. AZFa, AZFb, AZFc and partial deletions within the AZFc region were analyzed using multiplex PCR method. Finally, the association between paternal age and male infertility was evaluated. No AZFa, AZFb or AZFc deletions were found in the control group. Seven infertile men had deletions as the following: one AZFb, five AZFc, and one AZFab. Partial deletions of AZFc [gr/gr] in 9 of the 100 infertile men [9/100, 9%] and 1 partial AZFc deletions [gr/gr] in the control group [1/100, 1%] were observed. In addition, five b2/b3 deletions in five azoospermic subjects [5/100, 5%] and 2 partial AZFc deletions [b2/b3] in the control group [2/100, 2%] were identified. Moreover, the risk of male infertility was influenced by the paternal age. The results of this study suggested that the frequency of Y chromosome AZF microdeletions increased in subjects with severe spermatogenic failure and gr/gr deletion associated with spermatogenic failure

Association between XPD [Lys751G1n] polymorphism and lung cancer risk: a population-based study in Iran

People are usually susceptible to carcinogenic aromatic amines, present in cigarrette smoke and polluted environment, which can cause DNA damage. Therefore, maintenance of genomic DNA integrity is a direct result of proper function of DNA repair enzymes. Polymorphic diversity could affect the function of repair enzymes and thus augment the risk of different cancers. Xeroderma pigmentosum group D [XPD] gene encodes one of the most prominent repair enzymes and the polymorphisms of this gene are thought to be of importance in lung cancer risk. This gene encodes the helicase, which is a component of transcription factor IIH and an important part of the nucleotide excision repair system. Studies reveal that individuals with Lys751Gln polymorphism of XPD gene have a low repairing capacity to delete the damages of ultraviolet light among other XPD polymorphisms. In this case-control study, first Lys751Gln polymorphism was genotyped, then its association with lung cancer risk was analyzed. Genomic DNA was extracted from the whole blood sample of 640 individuals from Iran [352 healthy individuals and 288 patients]. Allele frequencies and heterozygosity of Lys751Gln polymorphism were determined using polymerase chain reaction-restriction fragment length polymorphism method. According to statistical analyses, lung cancer risk in individuals with Lys751Gln polymorphism [Odd Ratio=1.8, 95% Confidence Interval 0.848-3.819] is approximately twice as high as that of Lys/Lys genotype, however 751Gln/Gln genotype did not relate to lung cancer risk [Odd Ratio=0.7, 95% Confidence Interval 0/307-1/595]. This study suggests that heterozygous polymorphism [Lys/Gln] increases the sensitivity of lung cancer risk, while homozygous polymorphism [Lys/Lys] probably decreases its risk and C allele frequency shows no remarkable increase in the patients

CYP1B1 L432V polymorphism and lung cancer risk in the Iranian population

Lung cancer is considered as one of the most frequent cancers worldwide, and has been the cause of more than one million mortalities each year. Exposure to tobacco smoke is the primary cause of most lung cancers, since it contains several thousand compounds, including more than 50 known carcinogens. However, a small fraction of individuals who are exposed to tobacco smoke develop lung cancer, therefore genetic factors may render some tobacco smokers more susceptible to cancer. Genetic polymorphism in genes that encode metabolizing enzymes may be related to differentiated susceptibility of malignancy. CYP1B1 protein is a member of the more significant CYP1 subfamily enzymes, involved in environmental carcinogen metabolic activation. The most studied polymorphism in CYP1B1 gene includes 4325 C-G, resulting in an amino acid change from leucine to valine amino acid. A case-control study [included 65 lung cancer cases and 80 healthy controls] was designed based on the RFLP-PCR method to estimate the possible association of this polymorphism with lung cancer susceptibility in the Iranian population. Regarding the distribution of CYP1B1 L432V genotypes, there were no meaningful differences among controls and lung cancer patients, however among patients carrying the CC genotype, tobacco smokers had a considerable elevated risk for lung cancer compared to those who had the GG genotype. CYP1B1 L432V polymorphism has an important role in lung cancer risk. Therefore, further studies are recommended for investigation of other related CYP1B1 gene polymorphisms, their association with affective genes and regulatory factors in the Iranian population.

Relationship between XPD Lys 751 Gln polymorphism and colorectal cancer risk: a case-control study in a population-based study

In our study, we analyzed the allelic frequency of XPD Lys751Gln polymorphism of the XPD gene and the correlation between its variant alleles with colorectal cancer in patients and control groups. Human cells are routinely exposed to mutagenic and carcinogenic aromatic amines via smoking, pollution areas and other sources. These chemicals can form DNA adducts in vivo and thus lead to DNA damage. Amongst the known genetic polymorphisms of the DNA-repair genes the xeroderma pigmentosum group D [XPD, also known as ERCC2] has been the most extensively studied most commonly. This study has examined the relationship between the XPD Lys 751 Gin polymorphism and colorectal cancer in 88 patients and their 88 age and sex-matched controls. Genomic DNA from peripheral whole blood was extracted using Miller method to determine the genotype of subjects with RFLP-PCR analysis. This study shows cancer patients have more of the heterozygous genotype [XPD Lys 751 Gin] compared to control group. However the results are not statistically significant. Furthermore, colorectal cancer was less common in individuals with recessive homozygous genotype [P< 0.0001]. This study suggests that individuals with heterozygous polymorphism [Lys/Glri] may have an increased susceptibility to colorectal cancer compared to other polymorphisms [Lys/Lys and Gin/Gin]

Molecular analysis of the clavulanic acid regulatory gene isolated from an Iranian strain of streptomyces clavuligerus, PTCC 1709

The clavulanic acid regulatory gene [claR] is in the clavulanic acid biosynthetic gene cluster that encodes ClaR. This protein is a putative regulator of the late steps of clavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR, isolated from the Iranian strain of Streptomyces clavuligerus [S. clavuligerus]. In this experimental study, two different strains of S. clavuligerus were used [PTCC 1705 and DSM 738], of which there is no claR sequence record for strain PTCC 1705 in all three main gene banks. The specific designed primers were subjected to a few base modifications for introduction of the recognition sites of BamHI and ClaI. The claR gene was amplified by polymerase chain reaction [PCR] using DNA isolated from S. clavuligerus PTCC 1705. Nested-PCR, restriction fragment length polymorphism [PCR-RFLP], and sequencing were used for molecular analysis of the claR gene. The confirmed claR was subjected to double digestion with BamHI and ClaI. The cut claR was ligated into a pBluescript [pBs] vector and transformed into E. coli. The entire sequence of the isolated claR [Iranian strain] was identified. The presence of the recombinant vector in the transformed colonies was confirmed by the colony-PCR procedure. The correct structure of the recombinant vector, isolated from the transformed E. coli, was confirmed using gel electrophoresis, PCR, and double digestion with restriction enzymes. The constructed recombinant cassette, named pZSclaR, can be regarded as an appropriate tool for site directed mutagenesis and sub-cloning. At this time, claR has been cloned accompanied with its precisely selected promoter so it could be used in expression vectors. Hence the ClaR is known as a putative regulatory protein. The overproduced protein could also be used for other related investigations, such as a mobility shift assay

role of matrix metalloproteinase-3 functional 5A/6A promoter polymorphism in tumor cell progression and metastasis of breast cancer

In the human genome, chromosome 11 contains a cluster of matrix metalloproteinase [MMP] genes. Single nucleotide polymorphisms in the promoter region of MMP genes are important for MMP expression. A common adenine deletion polymorphism [5A] at position -1171 of the MMP-3 gene promoter [5-AAAAAACCAT-3 change to 5-AAAAACCAT-3] facilitates transcriptional factor binding and MMP-3 promoter activity. A case-control study was performed including 120 breast cancer patients [60 patients with metastatic activity and 60 patients without metastatic activity]; and 60 healthy controls. Whole blood samples were obtained from patients and healthy controls. Genomic DNA was extracted from samples and the MMP-3 5A/6A genotypes were determined using PCR-RFLP. MMP-3 genotype distributions between patients and controls were similar [OR= 0.89, 95%CI, 0.43-1.84, P= 0.047]. It was observed that the 5A allele was more frequent among patients with metastatic activity than controls [OR= 2.9, 95%CI, 0.94-8.9, P= 0.074]. Therefore, the 5A polymorphism in the MMP-3 promoter showed correlation with the metastasis group than patients without metastasis; both at the time of diagnosis. However our results do not show evidence for correlation between 5A/6A polymorphism and breast cancer susceptibility