Association of ACE I/D and AGTR1 A1166C Gene Polymorphisms and Risk of Uterine Leiomyoma: A Case-Control Study.

Objective: Uterine leiomyoma (UL) can be considered as the most common benign gynecological tumors of the smooth muscle cells in the myometrium. They are likely to be associated with infertility and recurrent abortion as well as obstructed labor and post-partum hemorrhage. Moreover, altered vascular-related genes can be linked to developing leiomyoma. Polymorphisms of the angiotensin-converting enzyme (ACE) gene are associated with some vascular diseases. The present study was carried out to investigate the association of ACE I/D and AGTR1 A1166C gene polymorphisms and the risk of uterine leiomyoma in a sample of Iranian population. Methods: The study was carried out on a total of 413 women divided into 202 patients with diagnosed uterine leiomyomas and a control group of 211. Genotyping was performed using the PCR or PCR-RFLP methods. Results: The ID and DD genotypes of ACE I/D polymorphism were associated with 2 and 2.9 fold higher risk of UL compared to II genotype (OR, 2 [95% CI, 1.3 to 3.2]; P = 0.004 and OR, 2.9 [95% CI, 1.6 to 5]; P = 0.0002). The frequencies of ACE D alleles were 53.7% in women with UL and 40.3% in controls, which were observed to be statistically different (P < 0.0001). The alleles and genotypes of AGTR1 A1166C polymorphism were not different between UL and control women (P=0.9). Conclusion: The ACE ID and DD genotypes were associated with a higher risk of UL. No relationship was found between AGTR1 A1166C polymorphism and UL.

Association of the placental VEGF promoter polymorphisms and VEGF mRNA expression with preeclampsia.

BACKGROUND: Preeclampsia (PE), is a pregnancy-specific complication with the placental origin which associated with altered expression of angiogenic factors. Vascular Endothelial Growth Factor (VEGF), is a growth and hypoxia-induced factor which contributes to the regulation of various processes. The present study has investigated the association of the placental VEGF -634G/C (rs2010963), -1154G/A (rs1570360), and -2549 I/D (18bpindel) polymorphisms in the promoter region with VEGF mRNA expression in PE women and control group. METHODS: This case-control study was performed on the placenta of 84 PE women and 103 controls after delivery. Genotyping of the VEGF polymorphisms was done by PCR or PCR, PCR-RFLP or sequencing methods. The mRNA expression levels were measured by Quantitative Real-Time PCR. RESULTS: The relative mRNA expression of VEGF gene was significantly higher in PE women compared to controls. The relative mRNA expression of VEGF gene was significantly higher in women with -634CC genotype compared to CG + GG genotypes in PE women and total studied women but not in control women. However, there was no association between the placental VEGF -1154G/A and -2549 I/D polymorphisms and VEGF mRNA expression neither in PE nor in control groups. CONCLUSION: The current study found higher mRNA expression of placental VEGF gene in PE women. The mRNA expression of the placental VEGF gene has been up-regulated in the placenta of women with -634CC genotype. No association was found between the placental VEGF -1154G/A and -2549 I/D polymorphisms and VEGF mRNA expression.

Antifungal effects of ethanolic and aqueous extracts of Vitexagnus-castus against vaginal isolates of Candida albicans.

BACKGROUND AND PURPOSE: Vulvovaginal candidiasis is one of the most common infections in female genital organs, which is caused by Candida species. Candida albicans is the causative agent of more than 80% of infections, and the role of non-Candida strains in the disease etiology is less prominent. The expansion of Azoles resistance among C. albicans strains is considered an important medical problem. According to previous studies, Vitex agnus-castus (vitex) has some antimicrobial effects. We aimed to evaluate the anti-fungal effects of aqueous and alcoholic extracts of vitex against clinical vaginal isolates of C. albicans in comparison with fluconazole. MATERIALS AND METHODS: Gas chromatography-mass spectrometry analysis was performed on vitex to identify its possible bioactive components. Forty C. albicans clinical isolates were identified by using germ tube, chlamydospore production, culture on CHROMagar, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Finally, after the extraction of vitex, drug susceptibility test was carried out according to the clinical laboratory standards institute (CLSI) M27-S4 document guidelines. RESULTS: The major chemical components of vitex leaf as determined by gas chromatography included α-Pinene, isoterpinolene, caryophyllene, and azulene. The minimum inhibitory concentrations (MICs) of aqueous and alcoholic extracts of vitex, as well as fluconazole were within the ranges of 15.62-62.5, 7.81-15.62, and 0.25-8 µg/mL, respectively. CONCLUSION: Our findings showed that the alcoholic and aqueous extracts of vitex had antifungal activity against clinical isolates of C. albicans. Moreover, the alcoholic extract of vitex and fluconazole were more effective against clinical vaginal isolates of C. albicans compared to the aqueous extract of vitex.

Association between ERα polymorphisms and systemic lupus erythematosus: susceptibility and in silico analysis.

BACKGROUND: Systemic lupus erythematous (SLE) is a multisystem and autoimmune disorder leading to damage of multi-organ systems. The current study aimed to assess the possible association between ERα gene polymorphisms and SLE in a southeast Iranian population. METHODS: The ERα PvuII and XbaI polymorphisms were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method in 170 SLE patients and 186 healthy subjects. RESULTS: There was no association between ERα PvuII and XbaI polymorphisms and SLE susceptibility; however, the combination of the TC/AA and CC/GG genotypes of ESR α PvuII and XbaI polymorphisms were more frequent in SLE patients. The results indicated that TT haplotype of the ERα gene polymorphisms could increase the SLE risk almost 2.4-fold (odds ratio 2.4, 95% CI 1.3-4.3, P = 0.005). The in silico analysis revealed that the ERα PvuII and XbaI single nucleotide polymorphisms occurred in acceptor splicing sites, and these mutations can lead to the increase of Human Splicing Finder score of the mutant alleles. CONCLUSIONS: The ESR α PvuII and XbaI polymorphisms have no association with SLE; however, the combination of the TC/AA and CC/GG genotypes were associated with SLE susceptibility.

Genetic variants in 3'-UTRs of MTHFR in the pregnancies complicated with preeclampsia and bioinformatics analysis.

Preeclampsia (PE) as a pregnancy-specific disorder is the major cause of mortality and morbidity of mothers and fetuses. This study attempts to investigate the possible association between the 2572C>A (rs4846049) and 4869C>G (rs1537514) polymorphisms in the 3'- untranslated region of MTHFR gene and the risk of PE. A total of 198 patients diagnosed with PE and 171 unrelated, age matched healthy pregnant women, were recruited for this case-control study. The MTHFR 2572C>A and 4869C>G genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The CG genotype of MTHFR 4869C>G was associated with decreased risk of PE, and this genotype was found to be a protective factor for PE susceptibility. There was no significant difference in the genotypes of MTHFR 2572C>A polymorphism between PE patients and control group. The frequency of combined AC/CG genotypes of MTHFR 2572C>A and 4869C>G polymorphisms were less frequent in PE patients and were associated with a lower risk of PE. The C-G and A-G haplotypes of MTHFR 2572C>A and 4869C>G polymorphisms were significantly lower in PE patients. In conclusion, the CG genotype of MTHFR 4869C>G polymorphism was associated with a lower risk of PE. No association was found between MTHFR 2572C>A polymorphism and PE.

Polymorphisms of the folate metabolizing enzymes: Association with SLE susceptibility and in silico analysis.

Systemic lupus erythematosus (SLE) is a typical autoimmune disorder with multiple organ involvement and unknown etiology. It has been shown that polymorphic variants of the genes encoding key enzymes of folate and methionine metabolism may influence DNA methylation. Genomic DNA was extracted from blood samples of 150 SLE patients and 160 controls, matching age, sex, and ethnicity. MTHFR rs1801133C>T and MTR rs1805087A>G polymorphisms were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. MTHFR rs1801131A>C genotype was determined by the tetra-primer amplification refractory mutation system (Tetra-ARMS), and DHFR rs70991108 polymorphisms' genotyping was performed by PCR methods. In-silico approach was used to analyses the effects of these variations on the structure of mRNA and protein. MTHFR rs1801131AC+CC genotypes were significantly higher in the SLE patients compared to the controls (37 vs. 26%, OR 1.7 (95% CI 1-2.8); p=0.03). The frequency of MTHFR rs1801131C allele was significantly higher in the SLE patients than the controls (22 vs. 15%, p=0.02). However, there was no association between MTHFR rs1801133C>T polymorphism and SLE. The frequency of CT haplotype of MTHFR rs1801133C>T and rs1801131A>C polymorphisms was significantly higher in the SLE patients (20 vs. 12%), and CT haplotype may be potentially a risk factor for SLE susceptibility [OR 1.9 (95% CI 1.2-2.9); p=0.006]. There was no association between alleles and genotypes of DHFR rs70991108 polymorphism and SLE susceptibility. The frequency of MTR rs1805087AG genotype was less frequent in the SLE patients compared to the controls, and this genotype could decrease the SLE risk (35 vs. 48%), (OR, 0.6 (95% CI, 0.4 to 0.9), p=0.03). In silico-analysis showed that both of MTHFR rs1801133C>T and rs1801131A>C SNPs made fundamental changes in the secondary structure of MTHFR-mRNA (p=0.0412 and p=0.1641; p<0.2). Also, structural analysis of the rs1801131A>C variation showed a significant effect on MTHFR function. Bioinformatics analysis showed that rs70991108 polymorphism in DHFR gene would lead to a significant alteration of the splicing process. In conclusion, MTHFR rs1801131 AC+CC genotypes could be a risk factor and MTR rs1805087AG genotype could be a protective factor for SLE susceptibility. There was no association between MTHFR rs1801133C>T and DHFR rs70991108 polymorphisms and SLE.

The placental vascular endothelial growth factor polymorphisms and preeclampsia/preeclampsia severity.

Preeclampsia (PE) is a serious pregnancy-specific condition, which originates from placenta and finishes after delivery. The present study has investigated the association between placental VEGF I/D (rs35569394), -1154G/A (rs1570360), and -634G/C(rs2010963) polymorphisms and maternal VEGF -2549 I/D (rs35569394) polymorphism with PE and PE severity. In this case-control study, the maternal blood of 217 women with PE and 210 normotensive pregnant women and the placenta of 84 PE women and 103 normotensive women were collected after delivery. Genotyping was done by PCR or PCR-RFLP methods. The maternal VEGF-2549I/D genotypes were not associated with PE or PE severity. The placental VEGF -2549 I/D genotypes were not associated with PE too; however; the placental VEGF-2549 DD genotype was statistically different between women with severe PE and mild PE or the controls. The placental VEGF -634GC and CC genotypes were significantly higher in PE women and associated with 2.6 and 2-fold higher risk of PE, respectively. The VEGF -634GC and CC genotypes were associated with PE severity. No association was found between placental VEGF -1154G/A polymorphism and PE or PE severity. The placental DGC haplotype of VEGF -2549 I/D, -1154G/A, and -634G/C polymorphisms was associated with 2.9-fold higher risk of PE. However, the placental IAG haplotype was associated with 0.3-fold lower risk of PE. In conclusion, the placental VEGF -2549 DD genotype was associated with severe PE and the placental -634GC and CC genotypes were associated with PE and severe PE. No association was found between VEGF -1154G/A polymorphism and PE or PE severity.

The -2549 insertion/deletion polymorphism of VEGF gene associated with uterine leiomyoma susceptibility in women from Southeastern Iran.

OBJECTIVES: Vascular endothelial growth factor (VEGF) is an important angiogenic factor that regulates angiogenesis and mediates sex steroid-induced cell growth. The present study investigated the association of VEGF gene-2578C/A (rs699947) and -2549 insertion/deletion polymorphisms in the promoter region of VEGF-A gene and uterine leiomyoma susceptibility in Southeast of Iran. MATERIAL AND METHODS: One hundred and fifty five women with uterine leiomyoma and 157 age, BMI, and ethnicity matched healthy women were enrolled in this study. VEGF gene -2578C/A polymorphism genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, and the -2549 insertion/dele-tion polymorphism was analyzed by PCR method. RESULTS: The frequency of alleles and genotypes of VEGF-2578C/A polymorphism was not different between women with uterine leiomyoma and the controls; however, a significant association was revealed between II genotype of -2549 insertion/deletion (I/D) polymorphism of VEGF gene and uterine leiomyoma. CONCLUSIONS: The findings showed that VEGF gene -2549 insertion/deletion polymorphism was associated with uterine leiomyoma.

Association of FAS A-670G Polymorphism and Risk of Uterine Leiomyoma in a Southeast Iranian Population.

BACKGROUND: Uterine leiomyoma (UL) is a benign tumor of uterine smooth muscle that affects women in reproductive ages. FAS has an important role in initial stages of apoptosis. Previous studies have shown an association between the FAS gene and tumorigenesis. In the present study, we evaluated the relationship between FAS A-670G (rs 1800682) and UL risk. METHODS: The FAS gene polymorphism of 155 women with UL and 157 healthy controls was analyzed by the polymerase chain reaction restriction fragment length polymorphism method. RESULTS: The AA, AG, and GG genotype frequencies of the FAS A-670G polymorphism were respectively 37.4, 42.6, and 20% in women with UL, and 46, 42.6, and 11.5% in healthy controls. The risk of UL in women was 1.5-fold greater in GG-genotype women than in AA-genotype women. The G allele frequencies were 41% in women with UL and 33% in healthy controls and statistically different (P = 0.03). CONCLUSION: The FAS polymorphism was associated with the risk of UL in a sample of Iranian women.