Association between Genetic Variants Linked to Premature Ovarian Insufficiency and Inflammatory Markers: A Cross-Sectional Study.

BACKGROUND: Premature menopause (PM) is the cessation of ovarian function before age 40. PM women are more likely to have cardiovascular diseases (CVDs), diabetes, and mental disorders. This is the first study that assessed the association of single nucleotide polymorphisms (SNPs) with anti-heat shock protein 27 (Hsp27), High-sensitivity C-reactive protein (hs- CRP), and PM and serum pro-oxidant-antioxidant balance (PAB), as putative risk factors for CVDs. We aimed to explore the association of oxidative stress markers with eight different SNPs shown to be related to premature menopause. MATERIALS AND METHODS: In this cross-sectional research, we included 183 healthy women and 117 premature menopausal women. We determined baseline characteristics for all participants and measured serum hs-CRP, anti-HSP-27 antibody titer, and PAB levels using the established methods. Genotyping for eight SNPs was done using the tetra amplification refractory mutation system polymerase chain reaction (Tetra-ARMS PCR) and allele-specific oligonucleotide PCR (ASO-PCR) methods. RESULTS: We found a significant difference between mean serum PAB levels and the genetic variant of rs16991615 (P=0.03). ANCOVA showed a significant effect of the genotypes rs4806660 and rs10183486 on hs-CRP serum levels in the case and control groups, respectively (P=0.04 and P=0.007). ANCOVA also showed an association between rs244715 genotypes and anti-hsp27 serum levels in the case group (P=0.02). There was a significant effect of the genotypes of rs451417 on the serum hs-CRP level in the control group (P=0.03). CONCLUSION: There was a significant association of the genetic variants related to PM with oxidative stress and inflammatory markers (serum PAB, anti-hsp27 antibody, and hs-CRP). Accordingly, this seems to be an effective approach to predicting susceptible subjects for cardiovascular and mental disorders as well as various cancers.

Micronutrients intake and genetic variants associated with premature ovarian insufficiency; MASHAD cohort study.

BACKGROUND AND AIM: premature ovarian insufficiency (POI) is defined as the menopause before 40 years of age, and its prevalence is reported to be two-fold higher in Iranian women than the average for woman globally. POI is associated with several cardio/cerebrovascular complications as well as an increased overall mortality. Genetic factors, and serum levels of minerals and vitamin D, have been reported to be related to the prevalence of POI. We have investigated the association between some POI -related genotypes with the serum levels of some important micronutrients. METHODS: One hundred and seventeen women with POI and 183 controls without any renal, hepatic, and thyroid abnormalities were recruited as part of the MASHAD study. Demographic and anthropometric features were recorded and blood samples were collected and processed. DNA was extracted from the buffy coat of blood samples from all participants and 8 POI-related single nucleotide polymorphisms (SNPs) were determined using ASO-PCR or Tetra ARMS-PCR. Serum minerals and vitamin D concentrations were measured using routine methods. RESULTS: In women with POI, serum copper, phosphate, and calcium were significantly different for those with rs244715, rs16991615, and rs4806660 genotypes, respectively. In our control population, significant differences were also found in serum copper concentrations between different genotypes of rs4806660, rs7246479, rs1046089, and rs2303369. After adjusting for all confounding factors, the women with POI carrying TC genotype (rs4806660) had a lower risk to have serum copper levels < 80 (µg/dL) than those carrying a TT genotype. Furthermore, women with POI carrying GG genotype (rs244715) had a 6-fold higher risk to have serum copper levels > 155 than those carrying AA genotype. CONCLUSION: The C and G alleles of the rs4806660 and rs244715 polymorphisms respectively are independently associated with serum copper in women with POI. Further studies are necessary to investigate the association of serum copper and other micronutrients in women and other POI -related polymorphisms.

The Effects of Lycopene on Modulating Oxidative Stress and Liver Enzymes Levels in Metabolic Syndrome Patients: A Randomised Clinical Trial.

OBJECTIVE: The pathogenesis of metabolic syndrome (MetS) complications involves the excessive production of
reactive oxygen species, inflammation, and endothelial dysfunction. Due to Lycopene, a highly unstable structure and
its significant effects on modulating the metabolic system, there is a strong need for a formula that can increase its
stability. The aim of this study was to develop an approach for encapsulating Lycopene and investigate its effects on
inflammatory markers, oxidative stress, and liver enzymes in patients with MetS.
Materials and Methods: This study is a simple randomized, double-blind, objective-based clinical trial that involved
eighty subjects with MetS, who were equally and randomly assigned to two groups: one group received 20 mg of
Lycopene per day for 8 weeks, and the Placebo group followed the same protocol as the Lycopene group but received
a placebo instead of Lycopene. They were called Lycopene and placebo, respectively. During follow-up visits after 4
and 8 weeks, 20 ml of blood was collected for evaluation of liver enzymes and some inflammatory related markers.
Results: Prior to the assignment of volunteers to their respective groups, there were no notable differences in C-reactive
protein (CRP), serum liver enzymes, systolic and diastolic blood pressure, or pro-oxidant-antioxidant balance (PAB)
between the Lycopene and placebo groups. However, our subsequent analysis revealed a significant reduction in the
serum levels of CRP (P=0.001) and PAB (P=0.004) in the group that received Lycopene. Our encapsulated Lycopene
treatment was not associated with a significant difference in serum levels of alanine aminotransferase (ALT), aspartate
transferase (AST), or alkaline phosphatase (ALP) between our two groups.
Conclusion: This study investigated the impact of Lycopene on individuals with MetS, revealing a noteworthy
modulation effect on PAB and inflammation linked to MetS. However, no significant differences was demonstrated in
serum levels of ALT, AST and ALP between the studied group (registration number: IRCT20130507013263N3).

The relationship between genetic variants associated with primary ovarian insufficiency and lipid profile in women recruited from MASHAD cohort study.

BACKGROUND AND AIM: Primary Ovarian Insufficiency (POI) is defined by the occurrence of menopause before the age of 40 years. It is often associated with cardiovascular disease (CVD). The purpose of this study was to explore the relationship between POI-associated genotypes cardiometabolic disorder risk factors. METHODS: One hundred seventeen women with POI and one hundred eighty-three healthy women without POI were recruited in this study. DNA was extracted and analyzed using ASO-PCR or Tetra ARMS-PCR. Lipid profiles were also assessed. RESULTS: Multivariate logistic regression analysis showed that individuals with GG vs. TT genotype of the rs1046089 SNP were more likely to have a higher serum LDL (p = 0.03) compared to the control group. There was also a significant association between low serum HDL and rs2303369 and rs4806660 SNP genotypes in the POI group. In the POI group, the percentage of those with high total cholesterol was lower in those with a CC genotype compared to those with a TT genotype (p = 0.03). CONCLUSION: Some SNPs reported to be associated with POI appear to be independently associated with dyslipidemia. These results may be helpful to identify subjects with POI who may be susceptible to CVD.

Genetic Determinants of Premature Menopause in A Mashhad Population Cohort.

BACKGROUND: Premature menopause is characterized by amenorrhea before age of 40 years, markedly raised serum luteinizing hormone (LH) level, follicle-stimulating hormone (FSH) level and reduced serum level of estradiol. Genome-wide analysis suggested several loci associated with premature menopause. Here, we aimed to analyze association of variants at the MCM8, FNDC4, PRRC2A, TLK1, ZNF346 and TMEM150B gene loci with premature menopause. MATERIALS AND METHODS: In this cross-sectional study, a total of 117 women with premature menopause were compared to 183 healthy women. Anthropometric indices were measured in all participants: height, weight, body mass index (BMI), waist circumference (WC) and wrist circumference. Eight single-nucleotide polymorphisms (SNPs) of the indicated genes (rs16991615, rs244715, rs451417, rs1046089, rs7246479, rs4806660, rs10183486 and rs2303369) were identified from the literature. Genotyping was performed using tetra-ARMS polymerase chain reaction (PCR) and ASO-PCR methods. RESULTS: T allele of the rs16991615, rs1046089, rs7246479 and rs10183486, C allele of rs244715, rs451417 and rs4806660 as well as TT genotype of rs2303369 were associated with an increased risk of premature menopause, likely causing susceptibility to primary ovarian insufficiency (POI) in comparison with C allele. We also found an association between the rs16991615 SNP with premature menopause. Frequency of the minor allele in cases was increased for all SNPs in comparison with controls. All minor alleles, except for rs2303369, showed a statistically significant increased odds ratio (OR). However, after Bonferroni correction for multiple testing, none of the P values were remained significant. CONCLUSION: The selected polymorphisms in MCM8, FNDC4, PRRC2A, TLK1, ZNF346 and TMEM150B genes may potentially affect susceptibility to premature menopause, although replication of the results in larger cohort could clarify this.

Prognostic Factors Associating with Pro-oxidant-antioxidant Balance; Neutrophils to Lymphocytes Ratio, Vitamin D, Heat Shock Protein 27, and Red Cell Distribution Width.

BACKGROUND: Several chronic diseases are mediated by oxidative stress. Oxidative stress affects cell morphology and function and is associated with alterations in the serum protein component. In the current study, we analyzed four individual prognostic factors associating with serum Pro-Oxidant-Antioxidant Balance (PAB): neutrophil to lymphocyte ratio (NLR), Vitamin D, anti-heat shock protein 27 (anti-hsp27) antibody titer, and red blood cell distribution width (RDW) to evaluate them as the potential prognostic markers. In the current study, we attempted to investigate the relationship between serum PAB, RDW, NLR, serum vitamin D and anti-hsp27 concentration. METHODS: A total of 852 participants (438 males and 414 females) aged 47.64 ± 7.77 years were recruited in a cross-sectional study based on the Mashhad stroke and heart atherosclerotic disorders (MASHAD) cohort study data. Hematological parameters, and vitamin D, PAB and anti-hsp27 antibody titers were measured using the Sysmex auto analyzer system and enzyme-linked immune sorbent assay (ELISA), respectively. RESULTS: The results showed a significant correlation between Vitamin D and anti-hsp27 antibody titers (r = -0.13 and p <0.001) as well as between RDW and serum PAB (r = 0.120 and p <0.001). Moreover, we found that serum PAB was positively associated with serum anti hsp27 antibody titers. The results showed increasing 1 unit of serum vitamin D can cause 3% decreases in anti hsp 27 values (OR = 0.97; CI 95% (0.96-0.99); p = 0.004). While this association was not significant for RDW, NLR and PAB (p >0.05) we found a significant association between serum PAB and serum anti hsp-27 antibody titers. Subjects with PAB levels 36.31-82.63 had a higher risk (1.83 fold) of having an increased anti-hsp27 antibody titers in comparison to the reference group (PAB level <36.31) (OR = 1.83 (95% CI = 1.33-2.52), p <0.001). CONCLUSION: The present study shows that serum vitamin D can be associated with reduction in inflammatory status probably by decreasing levels of serum anti-hsp27 antibody titers, reduction in oxidative stress and therefore may reduce the risk of cardiovascular diseases. Anti-hsp27 antibody titers are associated with oxidative stress through the serum PAB, therefore these factors may be of prognostic values in detecting oxidative stress and risk of atherosclerosis. The evaluation of these factors in a larger population may help further confirm these findings.