Nearmeen M. Rashad

Association of resistin gene polymorphisms with insulin resistance in Egyptian obese patients

BACKGROUND Obesity associated insulin resistance is a major risk factor for type 2 diabetes mellitus. Resistin is recently reported to provide a link between obesity, insulin resistance and type 2 diabetes mellitus. We aimed to investigate the possible associations of resistin gene (RETN) polymorphisms with obesity, and to detect whether these polymorphisms are associated with glucose intolerance and type 2 diabetes mellitus in obese patients. METHODS One hundred and forty-five Egyptian obese patients with or without glucose intolerance and 155 unrelated healthy controls were enrolled in this study. Polymorphisms of RETN +299G>A and RETN -420 C>G gene were detected by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP). Serum resistin was measured by ELISA. RESULTS RETN +299 AA and RETN -420 GG genotypes were significantly associated with obesity in Egyptian population. Moreover, the mutant alleles or genotypes of both examined polymorphisms were associated with impaired glucose tolerance and diabetes mellitus compared to normal glucose tolerant obese patients. Furthermore, our results revealed elevated waist/hip ratio, BMI, blood pressure, fasting blood glucose level, HOMA-IR, triglycerides, total cholesterol, resistin level, and decreased HDL cholesterol level in homozygote mutant genotypes carriers of both RETN polymorphisms among obese patients. CONCLUSION Resistin gene polymorphisms may play an important role in pathogenesis and susceptibility to obesity, impaired glucose tolerance, and type 2 diabetes mellitus in Egyptian population.

Genetic variation in the vitamin D receptor gene and vitamin D serum levels in Egyptian women with polycystic ovary syndrome

Obesity, insulin resistance, and hyperandrogenism are considered crucial parameters of polycystic ovary syndrome (PCOS) which might be related to vitamin D metabolism. The aim of this study was to investigate the associations between polymorphisms (TaqI and ApaI) in the vitamin D receptor gene (VDR) and PCOS among Egyptian women. We aimed also to elucidate the impact of these polymorphisms on vitamin D level, hormonal and metabolic parameters of PCOS. One hundred and fifty Egyptian women with PCOS and 150 unrelated controls were enrolled in this study. Polymorphisms of VDR Taq-I T/C (rs731236) and Apa-I A/C (rs7975232) gene were genotyped using polymerase chain reaction restriction fragment length polymorphism (PCR–RFLP). Serum 25 hydroxy vitamin D [25(OH) D] levels were measured by high-performance liquid chromatography. PCOS women had significantly lower levels of 25(OH) D compared to healthy women. Our results revealed that Taq-I CC genotype and C allele were associated with increased risk of PCOS, while the Apa-I polymorphism was not. Haplotype Taq-I C/ Apa-I C was associated with a higher PCOS risk more than controls. Moreover, there was a significant decrease of 25(OH) D levels in carriers of haplotype Taq-I C/ Apa-I C (with variant alleles) compared to the non-carriers. Results showed also that there was an obesity- VDR Taq-I genotypes interactions. These results suggested that, VDR Taq-I gene polymorphism is associated with increased risk of PCOS in Egyptian women.

Association between genes encoding components of the Leutinizing hormone/Luteinizing hormone–choriogonadotrophin receptor pathway and polycystic ovary syndrome in Egyptian women

Polycystic ovary syndrome (PCOS) is one of the most common endocrine–metabolic disorders; however, its pathophysiology is still unclear. Certain polymorphisms of luteinizing hormone beta‐subunit (LHβ) and LH/choriogonadotrophin receptor (LHCGR) genes may lead to changes in the bioactivity of this hormone. We aimed to investigate possible associations between polymorphisms in the LHβ and LHCGR genes and PCOS among Egyptian women. We also aimed to shed light on the impact of these polymorphisms on LH level, hormonal, and metabolic features of PCOS. A case–control study included unrelated 210 patients with PCOS and 200 healthy controls, and they were stratified according to their body mass index into two subgroups: lean and obese. Polymorphisms of LHβ G1502A and LHCGR [G935A, and ins18LQ] genes were genotyped using polymerase chain reaction–restriction fragment length polymorphism. Our results revealed that LHβ G1052A GA genotype and A allele, LHCGR G935A GA, AA genotypes, or A allele were significantly associated with PCOS risk, while the LHCGR ins18LQ polymorphism was not. Additionally, there is a synergism between LHβ G1052A minor A and minor A allele of LHCGR G935A or minor ins allele of LHCGR ins18LQ and susceptibility to PCOS. When we stratified PCOS women or controls into obese and lean subjects, we found that LHβ G1502A GA genotype and A allele being more frequent in the obese group when compared with lean patients with PCOS [The odds ratio and 95% confidence interval were 5.6 (1.30–24.56) and 5.15 (1.21–21.90), respectively, P = 0.01, for each group.] These results suggested that LHβ G1052A and LHCGR G935A genes polymorphisms are associated with increased risk of PCOS in Egyptian women especially in obese cases. There was a synergism between LHβ G1052A minor A allele and of LHCGR G935A minor A or minor ins alleles of LHCGR ins18LQ and PCOS risk.

Association between inflammatory biomarker serum procalcitonin and obesity in women with polycystic ovary syndrome.

Procalcitonin (PCT) is a potential biomarker of obesity-related, low-grade inflammation in polycystic ovary syndrome (PCOS). We aimed to investigate whether serum procalcitonin, high-sensitivity C-reactive protein (hs-CRP), white blood cell (WBC) and neutrophil counts are associated with polycystic ovary syndrome and with obesity. A case-control study included 107 women with PCOS and 93 healthy controls, they were then stratified according to their body mass index (BMI) into three subgroups; lean, overweight and obese. Serum PCT levels were measured using enzyme linked immunosorbent assay. PCOS patients had significantly higher levels of serum PCT, hs-CRP, WBC, and neutrophil counts than healthy women. In control and PCOS groups, serum PCT, hs-CRP levels, WBC, and neutrophil counts were significantly increased in overweight and obese women compared with lean subjects. Serum PCT levels were positively correlated with BMI, waist/hip ratio, total cholesterol, serum triglycerides, LH/FSH, hs-CRP values, WBC and neutrophil counts in PCOS women. We also observed that the increasing obesity was accompanied by a significant increase in the mean values of serum PCT and neutrophil counts in PCOS patients. We conclude that serum PCT is a novel biomarker for low-grade chronic inflammation in PCOS patients, especially in obese women. Thus, PCT is a promising useful marker for accurate diagnosis of the inflammatory activity of body fat and of PCOS.

Impact of insulin-like growth factor 2, insulin-like growth factor receptor 2, insulin receptor substrate 2 genes polymorphisms on susceptibility and clinicopathological features of hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma (HCC) is one of the major causes of cancer-related death worldwide. Insulin-like growth factor-2 (IGF-2) is an important autocrine and paracrine growth factor which may induce cell proliferation and inhibit cell apoptosis leading to the transformation of normal cells into malignant cells. This study aimed to evaluate the possible roles of IGF-2, insulin-like growth factor-2 receptor (IGF-2R), and insulin receptor substrate (IRS)-2 genes polymorphisms in susceptibility and clinicopathological features of HCC in Egyptian population. MATERIALS AND METHODS Four hundred and twenty-six HCC patients and 334 controls were enrolled in the study. Polymorphisms of IGF-2+3580, IGF-2+3123, IGF-2R 1619, and IRS-2 1057 gene were detected using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Serum IGF-2 were determined using ELISA. RESULTS Serum IGF-2 levels were significantly lower in HCC patients than in healthy controls. IGF-2+3580 AA genotype, IGF-2+3123 GG genotype or G allele, IRS-2 1057 DD genotype and D allele were significantly associated with HCC risk. The combination of IGF-2+3580 AA homozygosity and IGF-2R 1619 GG homozygosity presented a significant protective effect against HCC (OR=0.16,95% CI=0. 08-0.34, P=0. 005). Serum IGF-2 concentrations were significantly increased in HCC patients with the IGF-2+3580 AA genotype. We also observed that increased alpha-fetoprotein (AFP), Child-Pugh grade, tumor size, and number of malignant lesions were accompanied by a significant increase of serum IGF-2 mean values of in HCC patients. CONCLUSION IGF-2, IGF-2R, and IRS-2 genes polymorphisms and their combinations are associated with risk of HCC.

Lipocalin-2 expression and serum levels as early predictors of type 2 diabetes mellitus in obese women

Obesity and diabetes are increasing in epidemic proportions globally. Lipocalin‐2 (LCN‐2) is an inflammatory adipocytokine and obesity‐related marker of low‐grade inflammation. We aimed to investigate, for first time, the possible role of LCN‐2 expression and serum levels in prediction of impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM) among obese Egyptian women. This study included 188 obese women and 180 controls. Obese women were subdivided into three subgroups according to their fasting blood glucose, normal glucose tolerance (NGT), IGT and T2DM. Circulating LCN‐2 expression levels were determined by real time polymerase chain reaction. Serum LCN‐2 concentrations were assessed by ELISA. Our findings revealed that LCN‐2 expression and serum levels were higher in obese women compared to lean controls. They were higher in IGT and T2DM obese cases than in NGT obese women. Receiver operating characteristic analyses revealed that LCN‐2 expression level was a useful biomarker discriminating IGT from NGT and T2DM from IGT obese women (AUC were 0.735 and 0.740, respectively). It was an independent predictor of IGT and T2DM among obese women. Serum LCN‐2 level was a useful biomarker discriminating IGT from NGT and T2DM from IGT obese women (AUC were 0.705 and 0.728, respectively). It was independent predictor of T2DM without predicting IGT among obese women. The power of combined LCN‐2 serum levels and expression in discriminating between IGT from NGT and T2DM from IGT obese women was high (AUC = 0.717 and 0.741, respectively). In conclusion, LCN‐2 expression and serum levels could discriminate IGT from NGT and T2DM from IGT obese women and early predicting T2DM among obese women. While, LCN‐2 expression level was the independent predictor of IGT in obese women. Combination of both LCN‐2 expression and serum levels improved their diagnostic value in early detection of IGT and T2DM among obese women.

Angiopoietin-like protein 3 and 4 expression 4 and their serum levels in hepatocellular carcinoma

Background Hepatocellular carcinoma (HCC) is the 6th most common cancer and the 3rd leading cause of cancer causing death allover the world. The aim of this research to explore the clinical relevance of blood angiopoietin‐like protein‐3 (ANGPTL3) and ANGPTL4 expression and their proteins levels as non invasive biomarkers in cirrhotic and HCC patients and their influence on the clinicopathological features of HCC. Material and methods This work comprised 200 patients with chronic hepatitis (120 cases complicated with cirrhosis, 80 patients with primary HCC) and 100 controls. circulating ANGPTL3 and ANGPTL4 expression was estimated by real‐time polymerase chain reaction (RT‐PCR). ANGPTL3 and ANGPTL4 protein levels were determined by enzyme‐linked immunosorbent assay (ELISA). Results The circulating ANGPTL3 and ANGPTL 4 expression was significantly elevated in HCC cases compared to chronic hepatitis patients and controls. There were much more serum ANGPTL3 and ANGPTL4 values in HCC and chronic hepatitis patients as compared to controls, but we couldn’t detect this significance between chronic hepatitis and HCC cases as regards ANGPTL4. By Multiple stepwise linear regression analysis, an increased ANGPTL3 expression, alpha‐fetoprotein (AFP), serum ANGPTL 3 levels, Child–Pugh grade were significantly assosciatedassociated with increased risk of HCC. Logistic regression analysis revealed that ANGPTL 3 expression and AFP levels were the only pridectorspredictors of HCC (odds ratio (OR) = 8.9; 8.6 respectively, P = 0.003). Receiver operator characteristic (ROC) demonsterated that serum ANGPTL3 and ANGPTL4 levels were usufuluseful biomarkers discriminating chronic hepatitis cases from controls (AUC = 0.820,0.887, respectively P < 0.001). However, they fail to discriminate HCC patients from chronic hepatitis patients (P = 0.27,0.12 respectively). Moreover, ANGPTL3 and ANGPTL 4 expression were promising biomarkers discriminating chronic hepatitis cases from controls and those HCC cases from chronic hepatitis patients (P < 0.001). Combined ANGPTL3 expression and serum level wasn’t useful in discriminating HCC patient from chronic hepatitis (P = 0.09). In contrast, combined ANGPTL4 expression and serum level was an useful biomarker discriminating HCC cases from chronic hepatitis. Conclusion ANGPTL3 and ANGPTL 4 expression and serum levels can be promising non invasive biomarkers in diagnosis of chronic hepatitis and HCC especially their expression could be useful in discriminating HCC from chronic hepatitis patients. HighlightsANGPTL3 expression was elevated in HCC compared to chronic hepatitis.ANGPTL 4 expression was elevated in HCC compared to chronic hepatitis.Higher serum ANGPTL3 and ANGPTL4 values in HCC and chronic hepatitis.Increased ANGPTL3 expression, AFP, serum ANGPTL 3 were assosciatedassociated with HCC risk.ANGPTL 3 expression and AFP levels were the only pridectorspredictors of HCC.