Do Serum Nesfatin-1 Levels have A Predictive Role in Type-2 Diabetes Mellitus and its Microvascular Complications? A Case-Control Study.

Background Type 2 diabetes mellitus (T2DM) is a chronic disease with macrovascular and microvascular complications. Nesfatin-1 is a neuropeptide that develops from a more substantial intermediate compound known as nucleobindin 2 (NUCB2). Nesfatin-1 is known to play a role in regulating various physiological processes related to appetite, energy balance, and body weight. The purpose of the current study was to investigate the serum levels of nesfatin-1 in Egyptian patients with type 2 diabetes mellitus (T2DM) in comparison to healthy subjects and to assess the association of serum nesfatin-1 levels with the occurrence of diabetic microvascular complications in those patients. Methods This matched case-control study was conducted on 90 subjects 40-80 years old, with normal hepatic, cardiac, and respiratory functions, and 60 of them had T2DM. The included participants were divided into two groups: group 1, which was the control group and included 30 healthy subjects, and group 2, which included 60 subjects with T2DM. Group 2 was subdivided according to the presence or absence of microvascular complications into group 2a, which included 30 patients having T2DM with no microvascular complications, and group 2b, which included 30 patients having T2DM with one or more microvascular complications. Results T2DM patients had significantly lower serum nesfatin-1 levels (5.07±1.78 versus 9.05±2.1 mmol/L, <0.001) compared to healthy controls. Also, T2DM patients with microvascular complications had lower serum nesfatin-1 levels (4.32±1.72 versus 5.83±1.51 mmol/L, <0.001) compared to T2DM patients without microvascular complications. Serum nesfatin-1 level at a cutoff value of <8.09 mmol/L can be a marker for the detection of diabetes mellitus (DM) with the area under the curve (AUC) of 94.3%, 95% sensitivity, 74.3% specificity, 77.9% positive predictive value (PPV), and 65.7% negative predictive value (NPV), and at a cutoff value of <5.87 mmol/L can be a marker for the detection of microvascular complications of diabetes mellitus at AUC of 75.5%, 76.7% sensitivity, 67.3% specificity, 77.1% PPV, and 62.9% NPV. Conclusions Serum Nesfatin-1 may play a potential protective role in diabetes mellitus (DM) and its microvascular complications, as it decreases in individuals with diabetes and those with diabetic microvascular complications compared to controls. Additionally, serum Nesfatin-1 levels may have predictive value for the early detection of Type 2 diabetes mellitus (T2DM) patients, diabetic microvascular complications, and diabetic kidney disease (DKD) at cut-off values of < 8.09 (mmol/L), < 5.87 (mmol/L), and < 5.46 (mmol/L), respectively. Therefore, targeted Nesfatin-1 drug therapy may be tried to reduce morbidity and mortality caused by microvascular complications of diabetes.

Clinical significance of LncRNA-MIAT as a non-invasive diagnostic marker of nonHodgkin's lymphoma associated with occult hepatitis C virus infection

Aims: the current research aimed to investigate LncRNA-MIAT in patients with nonHodgkin lymphoma (NHL) and to assess its correlation with clinicopathological features and treatment protocols of NHLs among Egyptian patients with Occult hepatitis C virus (HCV) infection (OCI). Patients & Methods: This study was conducted on 20 patients with NHL and 30 healthy subjects as the control group. All subjects were screened for HCV-RNA in both plasma and PBMCs. RT-PCR determined lncRNA-MIAT. Results: lncRNA-MIAT relative expression level was upregulated in NHL groups (2.73±0.86) compared to controls (1.06±0.07), P ˂0.001*. Among NHL, patients with OCI (3.2±0.63) had significantly higher levels of lncRNA-MIAT compared to HCV (2.6±1.08) and non-HCV (2.4±0.4), P ˂0.001*. Additionally, the relative expression levels of lncRNA-MIAT were significantly positively correlated with laboratory and clinicopathological features of NHL. Interestingly, concerning the treatment of DLBCLNHL, there were significantly higher levels of lncRNA-MIAT in no treatment subgroup (n=10, 3.31±0.95) compared to successfully treated subgroups [CHOP (n=7, 1.58±0.34) and R-CHOP (n=3, 11.16±0.21), P ˂0.001* Conclusions: lncRNA-MIAT level was upregulated in NHL patients, particularly patients with OCI. Thus, circulatory lncRNA-MIAT may serve as a promising non-invasive diagnostic marker for NHL associated with OCI

GLP-1R polymorphism (rs1042044) and expression are associated with the risk of papillary thyroid cancer among the Egyptian population.

BACKGROUND: Glucagon like peptide-1 receptor (GLP-1R) agonist usage has previously been linked to an elevated incidence of thyroid cell adenomas and carcinomas in animals. AIM: The goal of this study was to determine if there was an association between GLP-1R gene polymorphism and expression with the risk of papillary thyroid carcinoma (PTC) and its clinical characteristics among the Egyptian population. MATERIAL AND METHODS: A total of eighty PTC patients and eighty healthy controls were included in the study. Real-time polymerase chain reaction (real-time PCR) and immunohistochemistry were used to determine GLP-1R expression in tumor tissue. The polymorphisms rs1042044 and rs6923761 in the GLP-1R gene were determined using PCR -restriction fragment length polymorphism (PCR-RFLP). RESULTS: PTC patients exhibited considerably greater frequencies of rs1042044 AA genotypes and A allele than controls (OR (95% CI) = 4.5 (1.75-11.8), P < 0.001; OR (95% CI) = 2.032 (1.301-3.17), P < 0.001 respectively). GLP-1R mRNA and protein expressions were higher in tumor samples than normal thyroid tissues among PTC patients. In addition, high GLP-1R expressions were more common in rs1042044 AA genotype carriers than CC carriers (P < 0.001). GLP-1R mRNA expression showed 95 % sensitivity and 97% specificity for PTC diagnosis. Moreover, GLP-1R expression was closely associated with LN metastasis, tumor size, tumor stage, and multifocality in PTC patients. CONCLUSION: This research provides new evidence linking the GLP-1R genetic polymorphism and tissue expression to PTC risk and invasiveness among the Egyptian population.

Osteoprotegerin expression and serum values in obese women with type 2 diabetes mellitus.

BACKGROUND: Obesity and diabetes prevalence are increasing worldwide. We aimed to detect the possible association of osteoprotegerin (OPG) gene expression with visceral adiposity indices and cardiometabolic risk factors among obese women. METHODS AND RESULTS: The study enrolled 150 controls and 150 obese cases subdivided into two subgroups non-diabetic (n = 70) and 80 patients with type 2 diabetes mellitus (T2DM). Circulating OPG gene expression levels were figured out by real time PCR (Polymerase Chain Reaction). Serum OPG levels were assessed by Enzyme Linked Immunosorbent Assay. Our results explored that OPG serum levels were lower in the obese women compared to control group (p < 0.001) and obese diabetics had higher serum levels of OPG in comparison to obese non-diabetic patients (p < 0.001). Expression levels of OPG were higher in obese women than controls (p < 0.001). Moreover, the blood expression levels of OPG gene were higher in diabetic obese patients than non-diabetics. We found positive correlations between parameters of metabolic syndrome and obesity indices. After adjustment of the traditional risk factors, stepwise linear regression analysis test revealed that OPG expression levels were independently correlated with glycated hemoglobin, high-density lipoprotein-cholesterol, and waist-to-hip ratio. CONCLUSIONS: OPG mRNA levels were associated with surrogate markers of insulin resistance in Egyptian obese women.

Interferon-gamma Expression Profile as Diagnostic Signatures of Unexplained Infertility in Female Patients Suffer from Hashimoto's Thyroiditis.

Diagnosis of unexplained infertility (UEI) is made by exclusion and a relatively common problem that affects couples worldwide. Unfortunately, it is a not uncommon for females to suffer from Hashimoto's thyroiditis (HT). Interferon-gamma (IFN- γ) has a central key role in HT and in the ability to conceive. We aimed to estimate serum IFN- γ level and its expression profile in Egyptian women with HT and assess their possible association with UEI. In this study, we examined 120 women with HT. We evaluated fertility in all patients; female patients who suffer from UEI were detected. Diagnosis of HT was based on the clinical data and the laboratory measures, enzyme-linked immunosorbent assay was used to measure serum IFN- γ, and the expression of IFN-γ messenger ribonucleic acid (mRNA) was assayed by real-time polymerase chain reaction (PCR). According to the results of this study, 37.5 % of the studied females who suffered from HT were diagnosed with UEI. The serum level of IFN-γ and its gene expression showed a significant positive correlation with thyroid-stimulating hormone (TSH) and thyroid autoantibodies. However, a negative correlation was found with anti-müllerian hormone (AMH), free T4 (FT3), and free T4 (FT4). Analysis by linear regression revealed that TSH and FT3 were associated with serum level of IFN-γ; while FT3 was associated with IFN-γ gene expression. We concluded that both are valued markers in diagnosing UEI in female patients suffering from HT.

The diagnostic significance of circulating lncRNA ADAMTS9-AS2 tumor biomarker in non-small cell lung cancer among the Egyptian population.

BACKGROUND: Long non-coding RNA ADAM metallopeptidase with thrombospondin type 1 motif, 9 antisense RNA 2 (ADAMTS9-AS2) was recognized as a novel tumor suppressor and plays an important role in the initiation and progression of malignant behavior in human cancers, although its plasma expression and clinical value in patients with non-small cell lung cancer (NSCLC) remain unknown. We aimed to analyze the diagnostic role of ADAMTS9-AS2 and cytokeratin 19 fragmentation antigen (CYFRA 21-1) in NSCLC. METHODS: The present study included 80 control subjects, 80 patients with benign lung lesion and 80 NSCLC patients. The expression of ADAMTS9-AS2 in the tissue and plasma was detected by a real-time polymerase chain reaction. Serum CYFRA 21-1 was analyzed using an enzyme-linked immunosorbent assay. RESULTS: In comparison with benign lung lesion and controls, tissue and plasma ADAMTS9-AS2 expression were significantly down-regulated in NSCLC (p < 0.001). Decreased ADAMTS9-AS2 expression was associated with TNM stages in NSCLC patients (p < 0.001). Up-regulation of CYFRA 21-1 was reported among NSCLC patients and it was associated with TNM staging. Tissue and plasma ADAMTS9-AS2 expression levels were the predicting factors for NSCLC and they both correlated negatively with CYFRA 21-1 levels. Plasma ADAMTS9-AS2 levels had a significant positive correlation with their tumor tissue levels. Plasma ADAMTS9-AS2 showed a higher sensitivity (95%) and specificity (99.1%) in the diagnosis of NSCLC than CYFRA 21-1 (61.3% sensitivity and 60% specificity). CONCLUSIONS: Our results suggested that decreased plasma ADAMTS9-AS2 expression might act as a novel non-invasive tumor biomarker in NSCLC diagnosis. Furthermore, plasma ADAMTS9-AS2 might predict aggressive tumor behavior.

Urinary exosomal microRNA-96-5p and microRNA-183-5p expression as potential biomarkers of bladder cancer.

Because of low sensitivity and specificity of the currently available urine biomarkers of bladder cancer (BC) detection and painful cystoscopy procedure. Our study aimed to evaluate expression of urinary exosomal miR-96-5p and miR-183-5p as probable non-invasive and accurate biomarkers for the diagnosis and follow up of BC. Using quantitative real-time polymerase chain reaction; expression of exosomal microRNA (miR)-96-5p and miR- 183-5p in the urine samples of 51 patients with BC, 21 patients with benign urinary bladder lesions and in 24 normal individuals as control group was done. Our study results showed higher expressions of both miR-96-5p and miR-183-5p in urine of BC patients in comparison with control group (P < 0.001 for each). Receiver operating characteristic curve (ROC) analysis showed that each microRNA had good sensitivity and specificity to differentiate BC from non-BC patients miR-96-5p 80.4% and 91.8% and miR-183-5p 78.4% and 81.6% respectively compared to cytology (37.3% and 100%). In addition, it was obvious that the sensitivity of combined miR-96-5p and miR-183-5p for the diagnosis of BC reached 88.2%% and specificity reached 87.8%, which were higher than each one alone. We also found that expression of miR-96-5p and miR-183-5p with high grade, and pathological stage was significantly increased. After surgery, collected urine samples showed significantly lower expression of miR-96-5p-: P < 0.001; and miR-183-5p: P = 0.002. In conclusion, urine miR-96-5p and miR-183-5p are promising tumor biomarkers of BC diagnosis; particularly, when they combined with each other or with urinary cytology.

Circulating miR-181a and miR-223 expression with the potential value of biomarkers for the diagnosis of systemic lupus erythematosus and predicting lupus nephritis.

BACKGROUND: MicroRNAs (miRNAs) contribute to the development and progression of systemic lupus erythematosus (SLE) by affecting a wide range of targeted genes and facilitating the development of lupus nephritis (LN). The present study aimed to analyze the serum expression of miR-181a and miR-223 in SLE patients and to assess whether they could serve as novel biomarkers for SLE diagnosis and to distinguish LN. METHODS: The study included 70 control subjects and 116 patients with SLE (67 non-LN and 49 LN groups). Circulating miR-181a and miR-223 expression levels were analyzed among the Egyptian population using a real-time polymerase chain reaction. RESULTS: Up-regulation of miR-181a was detected among SLE patients compared to healthy controls and higher values were reported among the LN group compared to the non-LN group. Down-regulation of miR-223 was reported among SLE patients compared to controls and lower values were reported among the LN group compared to the non-LN group. The higher miR-181a expression and the lower miR-223 expression were associated with higher stages of LN. SLE disease activity index, proteinuria and serum creatinine were independently correlated with miR-181a and miR-223 among SLE patients by linear regression analysis. Receiver-operating characteristic curve analysis revealed that combined miR-181a and miR-223 expression increased the sensitivity and specificity for the diagnosis of SLE and further distinguished LN from non-LN patients. CONCLUSIONS: miR-181a and miR-223 could play a role in evaluating SLE disease progression and prognosis. Combined miR-181a and miR-223 expression analysis could serve as novel serum-based biomarkers in the diagnosis of SLE and predicting LN among Egyptians.

Combined serum miR-29c and miR-149 expression analysis as diagnostic genetic markers for colorectal cancer.

Circulating miRNAs gathered much interest in cancer research as noninvasive biomarkers. The aim of this study was to analyze the expression of miR-29c and miR-149 among colorectal cancer (CRC) patients and to explore their diagnostic and prognostic potentials in relation to the clinical and pathological features. The expression levels of miR-29c and miR-149 were evaluated in the sera of 80 CRC patients, 80 colorectal adenoma (CRA) patients, and 80 healthy controls using quantitative real time polymerase chain reaction (PCR). Carcinoembryonic antigen serum levels were assayed using enzyme-linked immunosorbent assay. miR-29c and miR-149 were significantly downregulated among CRC patients compared with CRA and controls (miR-29c, 0.54 ± 0.19 vs. 0.86 ± 0.12, 0.99 ± 0.07, P < 0.001, respectively; miR-149, 0.46 ± 0.19 vs. 0.74 ± 0.012, 1.0 ± 0.22, P < 0.001, respectively). miR-29c and miR-149 significantly associated with advanced stages of CRC, tumor size, and lymphatic metastasis. By using receiver operating characteristic curve analysis, combined miR-29c and miR-149 revealed the highest diagnostic potential for CRA (area under the curve [AUC] = 0.967) from healthy controls as well as the diagnosis of CRC (AUC = 0.98) from CRA. Moreover, combined miRNAs revealed high diagnostic potential for the earlier stages of CRC compared with advanced stages (AUC = 0.96). In conclusion, combined serum miR-29c and miR-149 expression analysis established novel noninvasive biomarker for early CRC diagnosis.

The pattern of thyroiditis in multiple sclerosis: a cross-sectional study in a tertiary care hospital in Egypt.

BACKGROUND: Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system with varied clinical features. Disease-modifying drugs (DMDs) of MS associated with different types of thyroiditis. In this cross-sectional study, we aimed to assess the prevalence of thyroid dysfunction in MS and to investigate the association between DMDs and the risk of thyroiditis in MS. A cross-sectional study included 100 patients with relapsing-remitting multiple sclerosis (RRMS) in relapse, and the diagnosed was according to revised McDonald's criteria 2010. RESULTS: Our results revealed that the prevalence of thyroiditis was 40%; autoimmune (34%) and infective (6%) among patients with RRMS in relapse and cerebellar symptoms were significantly higher in patients with thyroiditis compared to patients without thyroiditis. Regarding the association between DMDs and thyroiditis, the prevalence of patients treated with interferon-beta-1b was higher in MS patients with thyroiditis compared to MS patients without thyroiditis. However, the prevalence of patients treated with interferon-beta-1a was lower in MS patients with thyroiditis compared to MS patients without thyroiditis. In addition, we found CMV infection was more common in patients treated by interferon beta-1b and candida infection was common in patients treated by fingolimod. CONCLUSIONS: Thyroiditis is commonly observed in patients with RRMS in relapse and higher prevalence of patients treated with interferon-beta-1b which is commonly associated with thyroiditis and CMV infection; however, candida thyroid infection was common in MS patients treated by fingolimod.

Association of vaspin gene expression and its serum level on the risk of ischemic stroke in type 2 diabetic Egyptian patients: Prospective case-control study.

We aimed to evaluate serum vaspin and its gene expression in patients with type 2 diabetes mellitus (T2DM) and to assess the association of serum vaspin and its gene expression with susceptibility of ischemic stroke (IS). The prospective case-control study included 50 healthy individuals in a control group, and 90 patients with and T2DM were stratified into two subgroups: patients with IS and patients without IS. The serum vaspin concentration was measured by enzyme-linked immunosorbent assay (ELISA). Real-time PCR was performed to detect the mRNA expression of vaspin. Serum vaspin and vaspin expression levels were significantly higher in IS compared to the non-IS group. Interestingly, they were positively correlated with other vascular and metabolic risks. Diastolic and systolic blood pressure, as well as hemoglobin A1c cholesterol (HbA1c), were independently correlated with serum vaspin. After adjusting for the traditional risk factors, the logistic regression analysis test was done to evaluate the predictor of IS among T2DM patients; the vaspin expression level was a statistical significance predictor of IS among T2DM patients. In conclusion, the higher levels of serum vaspin and vaspin expression levels in T2DM emphasizes the pivotal role of vaspin serum level and expression in the progression of metabolic and glucose abnormalities, thus, they could be used as biomarkers of IS.

Intercellular adhesion molecule-1 expression and serum levels as markers of pre-clinical atherosclerosis in polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder characterized by obesity, hyperandrogenism, and insulin resistance. Intercellular adhesion molecule-1 (ICAM-1) is a proinflammatory and proatherogenic cytokine which is associated with atherosclerosis, insulin resistance, and cardiovascular disease (CVD). The pathogenesis of PCOS is not precisely known. Thus, the purpose of this study was to investigate the potential role of ICAM-1 expression and serum ICAM-1 concentrations in pathogenesis of PCOS. Moreover, we aimed to evaluate the possible relationship between ICAM-1 gene expression with carotid intima-media thickness as well as clinic-morphological features of PCOS. METHODS: This case control study enrolled 180 patients with PCOS and 120 controls groups and they were stratified according to their fasting plasma glucose (FPG) into three subgroups; normal glucose tolerance (NGT) [n = 75], those with impaired glucose tolerance (IGT) [n = 65], and 40 patients with type 2 diabetes mellitus (T2DM). Circulating ICAM-1 expression levels were determined by real time polymerase chain reaction (RT-PCR). Serum ICAM-1 concentrations were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Our results revealed that PCOS patients had higher values of ICAM-1expression and serum levels. Among PCOS patients, T2DM patients had the highest values of ICAM-1 expression and serum levels compared to IGT and NGT subgroups. The ICAM-1 expression and serum levels were significantly positive correlated with cardiovascular risk and PCOS phenotypes. Linear regression test showed that HOMA-IR was the main predictors of serum ICAM-1 levels in PCOS. Receiver operating characteristic curve (ROC) analysis revealed that, the power of ICAM-1 expression levels was higher than serum ICAM-1 in diagnosis of PCOS and in differentiating T2DM from IGT and NGT subgroups. Interestingly, combination of both ICAM-1 expression and serum levels improved the diagnostic role of serum ICAM-1. CONCLUSION: ICAM-1 expression and serum levels were higher in women with PCOS compared to control group also, there was a strong independent association between higher ICAM-1 expression and serum levels with cardiovascular risks in PCOS group.

Serum Levels of Transforming Growth Factor Beta -1 (TGF-ß1) as An Early No Invasive Marker for Diagnosis of Lupus Nephritis in Systemic Lupus Erythematosus Patients.

About 40-50% of all patients with systemic lupus erythematosus (SLE) patients are associated with significant morbidity and a poor prognosis. The transforming growth factor ß-1(TGF-ß1) is a member of cytokines families which has emerged as an important player in the pathogenesis of autoimmune diseases, including SLE. In this study we aimed to evaluate TGF-ß1 as a noninvasive diagnostic test for early diagnosis of LN and to assess the correlations between TGFß-1 and clinic-pathologic characteristics as well as disease activity of SLE. This case-control study included 188 patients with SLE, stratified into two subgroups LN group and Non-LN group. We assessed diseases activity by SLE disease activity index and measured TGEß-1 by using ELISA. Our results showed that LN patients had significant lower values of serum TGF-ß1 compared with non-LN patients (P < 0.001). Moreover, there were significant differences between LN histopathological classes. The lowest levels values of serum TGFß1 was in Class V. There were significant negative correlations between levels of TGF-ß1 and SLEDAI, fever, arthritis, proteinuria, hematuria, serum creatinine, thrombocytopenia, lymphopenia, ESR, ANA, pus cell and cellular cast's, all (P < 0.01). In lupus nephritis patients, TGF-ß1 levels were positively correlated with eGFR, C3 and C4 (P < 0.001). Linear regression analysis revealed that, eGFR, CRP, thrombocytopenia, and serum creatinine were independently correlated with TGF-ß1 among lupus nephritis patients (P < 0.001). According to Receiver Operating Characteristic analysis, the sensitivity and specificity of TGF-ß1 were 91% and 65.5%, respectively in the diagnosis of LN among SLE patients. As LN group had significantly lower values of serum TGFß1 and the values further decreased with more damage of kidney tissues and progression of SLE activity. We conclude that serum TGF- ß1 could be a valuable non-invasive marker for assessment of LN activity and organ damage.

Kisspeptin and body weight homeostasis in relation to phenotypic features of polycystic ovary syndrome; metabolic regulation of reproduction.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a heterogeneous disorder characterized by a diverse collection of reproductive and metabolic abnormalities. kisspeptin (KISS) is novel peptides associated with regulation of metabolism, food intake, puberty and reproduction. The aim of the present study was to estimate KISS level in patients with PCOS, and to evaluate the possible relationship between KISS level with anthropometric measures as well as clinic-morphological features of PCOS. MATERIALS AND METHODS: cross section control study enrolled 90 control group and 105 patients with PCOS and they were stratified according to their body mass index (BMI) to; underweight (n = 9, BMI ˂19), normal weight (n = 25, BMI = 19.1-25), over weight (n = 34,BMI = 25.1-30), obese grade I (n = 12, BMI = 30.1-35) , obese grade II (n = 13, BMI 35.1-40) and obese grade III (n = 12, BMI˃40).Circulating KISS levels were measured using ELISA. RESULTS: Our results revealed that, KISS levels were higher in PCOS patients compared to controls. Among PCOS group, there were significant lower level of KISS levels in underweight, overweight and obese compared to normal weight group. Even more importantly, KISS levels decreased with increasing of BMI as the following, grade I, grade II and grade III. Moreover, it was negatively correlated to anthropometric measures, glycemic, lipid profile and positively correlated the phenotype characteristics of PCOS. Linear regression test observed that hirsutism score, HOMA-IR and LH were the main predictors of KISS levels in PCOS. CONCLUSION: circulating KISS is an important regulator of body weight and reproduction especially in PCOS women.

Effect of a 24-week weight management program on serum leptin level in correlation to anthropometric measures in obese female: A randomized controlled clinical trial.

BACKGROUND: Obesity is a major contributor to preventable disease and death across the globe. Obesity is complex. Although its risk factors are myriad and compounding, there is an urgent need for a deeper understanding of the way risk factors interact with each other. Leptin is a peptide regulates food intake and body weight. However, the notion of leptin as an anti-obesity hormone was called into question because obesity is typically associated with high leptin levels and not leptin deficiency thus, we aimed to measure leptin levels in obese female in correlation to anthropometric measures and to evaluate the impact of weight loss on its level and metabolic parameters. SUBJECT AND METHODS: case-control study enrolled 40 control groups, 50 obese women. We measured anthropometric measures BMI, Waist/hip ratio (WHR). Fat mass index (FMI%) and free fat mass index (FFMI%) were assessed by dual energy X-Ray absorptiometry (DEXA) The serum levels of leptin were measured by ELISA. RESULTS: Our results revealed that serum leptin levels were higher in obese women compared to controls. Moreover, it was positively correlated to anthropometric measures, glycemic and lipid profile. Linear regression analysis revealed that BMI was the main independent studied parameters associated with serum leptin level among other clinical and laboratory biomarkers. Interestingly, after 12 weeks of following the Mediterranean diet (MD)-based weight loss program, serum leptin levels were decreased. Logistic regression analysis was performed to detect the main predictors' biomarkers associated with weight loss among obese women. We found that serum leptin and FMI% were an independent predictor of response with odds ratios of 1.69 and 1.64 respectively (P < 0.001), Receiver operating characteristic analyses revealed that the AUC of serum leptin in discriminating obese women from lean ones was 0.893 (95% CI = 0.815-0.917) with sensitivity = 90%, specificity = 96%, and the cutoff values was 36.32 ng/ml. CONCLUSION: Serum leptin could be a valuable diagnostic marker of obesity and its comorbidities. Moreover, significant weight loss leads to decrease serum leptin levels and improvement of glycemic and lipid profiles.

Association of miRNA - 320 expression level and its target gene endothelin-1 with the susceptibility and clinical features of polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder characterized by obesity, hyperandrogenism, and insulin resistance (IR). MicroRNAs (miRNAs) are small noncoding RNA associated with ovarian follicle development and female fertility. The objective of this study was to investigate the role of miRNA- 320 and its target gene endothelin-1 (ET-1) as a noninvasive biomarker of PCOS and to evaluate its possible relationship with IR as well as clinic-morphological features of PCOS. METHODS: Case-control study enrolled 60 patients with PCOS and 40 control group. We subdivided our PCOS women according to homeostasis model assessments of insulin resistance (HOMA-IR) to PCOS women with and without IR.ET-1 levels were measured by ELISA. We estimated the serum expression level of miRNA- 320 by real-time polymerase chain reaction. RESULTS: Our results revealed that serum miR-320 expression level was lower in PCOS patients compared to controls, in particular, PCOS women with IR. Moreover, it was negatively correlated to its target gene; ET-I as well as fasting serum insulin (FSI), HOMA-IR, PCOS phenotype; hirsutism score, ovarian volume and antral follicle count (AFC). In the PCOS group, linear regression analysis revealed that only hirsutism and HOMA-IR was the main predictor of expression levels of miRNA - 320 among other clinical and laboratory biomarkers of PCOS. The sensitivity and specificity of serum miR-320 expression levels in diagnosis PCOS was 80, and 97.5% respectively. CONCLUSION: The Expression serum levels of miR-320 were lower in PCOS compared to control and it could be a noninvasive diagnostic biomarker of PCOS.

Serum miRNA-27a and miRNA-18b as potential predictive biomarkers of hepatitis C virus-associated hepatocellular carcinoma.

Hepatitis C virus (HCV) infection remains the main risk factor for chronic hepatitis (CHC), liver cirrhosis, and hepatocellular carcinoma (HCC). Changes in microRNA (miRNA) profiles can be associated with HCV infection and may either favor or inhibit the virus and/or its complication. Moreover, miRNAs have emerged as key regulators of various cancers including HCC. The aim of this work was to investigate the potentail role of miRNA-27a and miRNA-18b expression levels as non-invasive predictive biomarkers of hepatitis C virus-associated HCC. Furthermore, we aimed to explore potential association of these miRNAs expressions with HCC clinicopathological features' in Egyptian cases. This case control study included 200 participants [60 CHC patients, 39 post-HCV cirrhosis patients, 51 HCC cases], and 50 healthy volunteers. The serum miRNA-27a and miRNA-18b expression profiles were measured using quantitative real time-polymerase chain reaction (qRT-PCR). miRNA-27a and miRNA-18b expression levels were significantly increased in post-hepatitis C cirrhosis cases compared to control and CHC groups. In HCC group, only miRNA-27a expression levels were significantly increased. Moreover, miRNA-27a and miRNA-18b expression levels were positively correlated with distant metastasis, Child-Pugh grade, and lymph node metastasis. Logistic regression analysis revealed that miRNA-27a expression was an independent predictor of cirrhosis among CHC. Receiver operating characteristic (ROC) analyses showed that miRNA-27a and miRNA-18b expression levels were useful biomarkers discriminating cirrhosis from CHC (AUC were 0.672 and 0.487, respectively), and in differentiating HCC from post-hepatitis C cirrhosis (AUC were 0.897 and 0.723, respectively). By combined ROC analysis, power of miRNA-27a and miRNA-18b expression levels as discriminator between HCC from post-hepatitis C cirrhosis was high (AUC = 0.0.821). Serum microRNA-27a and miRNA-18b expression levels are promising diagnostic and non-invasive biomarkers of CHC, post-CHC cirrhosis, and HCC.

Interleukin-6 and hs-CRP as Early Diagnostic Biomarkers for Obesity-Related Peripheral Polyneuropathy in Non-Diabetic Patients.

The number of obese individuals is surging in developed and developing countries. Obesity predisposes to many serious illnesses. Peripheral neuropathy (PN) is one of its major complications. Interleukin-6 (IL-6) is a pro-inflammatory cytokine secreted by the adipose tissue and is believed to play a principal role in obesity complications. Meanwhile, hs-CRP is a known inflammatory biomarker. This study was designed to detect the role of serum IL-6 and hs-CRP as inflammatory biomarkers in early diagnosis of peripheral neuropathy (PN) in non-diabetic obese patients. Additionally, we aimed to explore the association between IL-6 with the clinical and electrophysiological tests of PN. This cross-sectional controlled study enrolled 150 obese patients and 50 subjects as a control group. The obese groups were subclassified according to BMI into three groups; all participants were subjected to a complete neurological examination and motor, and sensory peripheral nerve conduction studies. Serum IL-6 and hs-CRP levels were assessed using a commercially available ELISA. Our results revealed that obese patients with PN had statistically significant higher values of IL-6 and hs-CRP compared to obese patients without PN and controls (P < 0.001). Meanwhile, levels of both marker were inversely related to nerve conduction velocities (P < 0.001). Obese patients with PN had higher values of serum IL-6 and hs-CRP than diabetic patients without DPN. Because the diagnostic power of both markers in serum was significant, we conclude that they could be a useful diagnostic biomarker of PN.

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 (odd's 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.

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. © 2017 IUBMB Life, 69(2):88-97, 2017.