Plasma-based proteomic profiling identifies the distinct regulation of proteins in hyperplasia and endometrial cancer.

BACKGROUND: Among gynaecological malignancies, endometrial cancer (EC) is the most prevalent type of uterine cancer affecting women. This study explored the proteomic profiles of plasma samples obtained from EC patients, those with hyperplasia (Hy), and a control group (CO). A combination of techniques, such as 2D-DIGE, mass spectrometry, and bioinformatics, including pathway analysis, was used to identify proteins with modified expression levels, biomarkers and their associated metabolic pathways in these groups. METHODS: Thirty-four patients, categorized into three groups-10 with EC, 12 with Hy, and 12 CO-between the ages of 46 and 75 years old were included in the study. Untargeted proteomic analysis was carried out using two-dimensional difference in gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). RESULTS: In all three groups, 114 proteins that were significantly (p ≤ 0.05 and fold change ≥ 1.5) altered were successfully identified using peptide mass fingerprints (PMFs). Compared with those in the control group (CO), the EC samples had 85 differentially expressed proteins (39 upregulated and 46 downregulated), and in the Hy group, 81 proteins were dysregulated (40 upregulated and 41 downregulated) compared to those in the CO group, while 33 proteins exhibited differential regulation (12 upregulated and 21 downregulated) in the EC plasma samples compared to those in the Hy group. Vitamin D binding protein and complement C3 distinguished Hy and EC from CO with the greatest changes in expression. Among the differentially expressed proteins identified, enzymes with catalytic activity represented the largest group (42.9%). In terms of biological processes, most of the proteins were involved in cellular processes (28.8%), followed by metabolic processes (16.7%). STRING analysis for protein interactions revealed that the significantly differentially abundant proteins in the three groups are involved in three main biological processes: signalling of complement and coagulation cascades, regulation of insulin-like growth factor (IGF) transport and uptake by insulin-like growth factor binding proteins (IGFBPs), and plasma lipoprotein assembly, remodelling, and clearance. CONCLUSION: The identified plasma protein markers have the potential to serve as biomarkers for differentiating between EC and Hy, as well as for early diagnosis and monitoring of cancer progression.

Plasma Proteomic Signature of Endometrial Cancer in Patients with Diabetes.

The incidence and mortality of endometrial cancer (EC) have increased in recent years. There is mounting evidence that diabetes may play a role in the greater incidence of EC. The molecular mechanisms of the interaction between type 2 diabetes and EC are not yet clearly understood yet. The present study was undertaken to investigate the plasma proteomics of EC patients with diabetes in comparison to those of EC patients without diabetes. Plasma samples were obtained from age-matched patients (EC diabetic and EC nondiabetic). Untargeted proteomic analysis was carried out using a two-dimensional differential gel electrophoresis coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Of the 33 proteins identified, which significantly differed in the plasma abundance between groups, 17 were upregulated and 16 were downregulated. The majority of the altered proteins are involved in the acute phase reaction, cholesterol metabolism, scavenging of heme from plasma, and plasma lipoprotein assembly and mobilization. α-2-macroglobulin, Ras association domain-containing protein 3, apolipoprotein A-I, α-1B-glycoprotein, and zinc-α-2-glycoprotein were significantly upregulated. The significantly downregulated proteins included haptoglobin, apolipoprotein A-IV, hemopexin, and α-1-antichymotrypsin. The differential expression of proteins found in patients who had EC and diabetes indicated severe disease and a poor prognosis. The protein interaction analysis showed dysregulation of cholesterol metabolism and heme scavenging pathways in these patients.

Metabolomic Profiling of Blood Plasma in Females with Hyperplasia and Endometrial Cancer.

Uterine cancer is the most prevalent gynecologic malignancy in women worldwide. Endometrial cancer (EC) has an 81% five-year survival rate, depending on disease stage and time of diagnosis. While endometrial cancer is largely treatable when detected early, no established screening techniques are available in clinical practice. As a result, one of the most significant issues in the medical field is the development of novel ways for early cancer identification, which could boost treatment success rates. Liquid chromatography-high-resolution mass spectrometry (LC-HRMS)-based metabolomics was employed to explore the metabolomic markers and pathways unique to this cancer type and link them to the benign endometrial hyperplasia that may progress to cancer in 5% to 25% of patients. The study involved 59 postmenopausal participants, 20 with EC type 1, 20 with benign hyperplasia, and 19 healthy participants. Metabolite distribution changes were analyzed, and 338 of these features were dysregulated and significant. The first two main components, PC1 and PC2, were responsible for 11.5% and 12.2% of the total metabolites, respectively. Compared with the control group (CO), EC samples had 203 differentially expressed metabolites (180 upregulated and 23 downregulated); in hyperplasia (HP), 157 metabolites were dysregulated (127 upregulated and 30 downregulated) compared to the CO group while 21 metabolites exhibited differential regulation (16 upregulated and 5 downregulated) in EC plasma samples compared to the HP group. Hyperplasia samples exhibited similar metabolic changes to those reported in cancer, except for alterations in triglyceride levels, 7a,12 b-dihydroxy-5b-Cholan-24-oic acid, and Hept-2-enedioyl carnitine levels. The metabolites N-heptanoyl glycine and -(Methylthio)-2,3-isopentyl phosphate and formimino glutamic acid can be specific markers for hyperplasia conditions and dimethyl phosphatidyl ethanolamine and 8-isoprostaglandin E2 can be specific markers for EC conditions. Metabolic activities rely on mitochondrial oxidative phosphorylation for energy generation. The changes in metabolites identified in our study indicate that endometrial cancer cells adopt alternative strategies to increase energy production to meet the energy demand, thereby supporting proliferation.

Urinary Proteome Differences in Patients with Type 2 Diabetes Pre and Post Liraglutide Treatment.

Diabetes mellitus is a chronic multisystem disease with a high global prevalence. The glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide is known to lower glucose levels and reduce weight. However, the mechanisms underlying the benefits of liraglutide treatment in patients with type 2 diabetes mellitus (T2DM) remain unclear. Twelve male patients with T2DM (pre and post liraglutide treatment) and HbA1c between 8% and 11% were recruited. In the present study, a two-dimensional difference gel electrophoresis (2D-DIGE) matrix-assisted laser desorption/ionization-time of flight (MALDI TOF) mass spectrometric approach combined with bioinformatics and network pathway analysis was used to explore the urine proteomic profile. The mean age of the patients was 52.4 ± 7.5 years. After treatment with liraglutide, a statistically significant change (p < 0.006) was observed in HbA1c with no significant changes in body weight or markers of dyslipidemia. Two-dimensional difference gel electrophoresis identified significant changes (≥1.5-fold change, ANOVA, p ≤ 0.05) in 32 proteins (4 down- and 28 upregulated) in liraglutide post treatment compared to the pre-treatment state. Albumin, serotransferrin, metallothionein-2 (MT-2), and keratins K1 and K10 were found to be upregulated after liraglutide treatment. The patients showed significant improvement in glycemic control after the 12-week treatment with liraglutide. The renoprotective effect of liraglutide may be linked to the increased urinary abundance of MT-2 and the decreased abundance of zinc alpha 2-glycoprotein (ZAG) and Alpha-1 antitrypsin (α1-AT). More studies are needed to elucidate the molecular mechanisms behind the renoprotective effects of liraglutide.

Cytokines Involved in COVID-19 Patients with Diabetes: A Systematic Review.

BACKGROUND: Hyperglycemic condition and compromised immune system may contribute to the progression of COVID-19 infection and increase the disease severity, relatively requiring a longer recovery period among diabetic patients. OBJECTIVE: A systematic review was conducted to examine cytokine levels, the prevalence of risk factors, and other comorbidities in COVID-19 patients with and without diabetes mellitus during the early COVID-19 outbreak. METHODS: A systematic literature search was conducted in PubMed central, PMC Europe databases, and Web of Science, evaluating the articles published between Dec 1st, 2019, and June 15th, 2020. This systematic review was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). RESULTS: The systematic search generated 14,960 articles and ended up with 9 articles, of which 8 articles involved data on the comparison of cytokines in diabetic and non-diabetic subjects with COVID-19, while 4 of them involved data on cytokines in the diabetes patients compared either by the severity of diseases or the rate of survival. Among the studied cytokines, interleukin-6, interleukin- 8, and tumor necrosis factor-α may cause the worst prognosis or fatality among diabetic patients. Increased cytokine levels indicate higher mortality and are linked to risk factors and comorbidities, such as hypertension and cardiovascular disease. Management of diabetes by insulin treatment may reduce the rate of mortality among diabetic patients but may be contraindicated in diabetic patients with COVID-19 who had at least one previous comorbidity, especially hypertension and CVD. CONCLUSION: The pathophysiological mechanisms linked to cytokine storm in diabetic patients may lead to the design of treatment strategies in the future, thus improving early diagnosis of the disease and mitigating cytokine storm-associated morbidity and mortality.

Association of sex hormone binding globulin with gestational age and parity in gestational diabetes mellitus.

OBJECTIVE: Recent research on sex hormone binding globulin (SHBG) has emphasized its role in the prediction of gestational diabetes mellitus (GDM) development. SHBG is associated with glucose tolerance status, and its level is regulated by prenatal and perinatal factors during pregnancy. This study aimed to determine the levels of SHBG in pregnant women with normal glucose tolerance and those with GDM in association with parity and gestational age (weeks). MATERIALS AND METHODS: This cross-sectional study enrolled 218 pregnant women (165 controls and 53 women with GDM). Serum SHBG levels were analyzed using enzyme-linked immunosorbent assay. The association of SHBG with gestational age was assessed using multivariate regression analysis after adjustment for GDM-related risk factors. RESULTS: Parity sub-group analyses indicated the presence of significant differences in the SHBG levels between nulliparous women in the GDM and control groups (p = .04). Moreover, in the GDM group, SHBG was significantly associated with gestational age beyond the risk factors of GDM. CONCLUSION: This study demonstrated a strong association between SHBG and gestational age in women with GDM. Our findings suggest that parity and gestational age should be considered in the analysis of SHBG as a marker for GDM diagnosis.

Association of urinary non-albumin protein with the different urinary marker for glomerular and tubular damage in patients with type 2 diabetes.

BACKGROUND/AIM: In recent years, the diagnostic utility of urinary protein levels has been demonstrated for the early detection and progression of kidney disease. This study aimed to evaluate the associations of the non-albumin protein (NAP) with different urinary marker for tubular and glomerular damage in patients with type 2 diabetes (T2D). METHODS: In this observational cross-sectional study, 424 patients with T2D duration > 10 years were classified into two groups according to estimated glomerular filtration rate (eGFR). The ratios of different urinary markers (albumin, NAP, total protein, transferrin, retinol-binding protein (RBP), and neutrophil gelatinase-associated lipocalin (NGAL) to creatinine were analyzed. RESULTS: The levels of urinary biomarkers increased significantly with decrease in eGFR levels. In the group with moderately decreased eGFR, the albumin to-creatinine ratio (ACR), non-albumin protein-to-creatinine ratio (NAPCR), and total protein-to-creatinine ratio (PCR) were independently associated with all urinary markers after being adjusted for risk factors. The area under the receiver operating characteristics (ROC) curve for ACR and PCR had a better diagnostic value than other urinary biomarkers. Comparing ROC curve of NAPCR with other urinary biomarkers, it was significantly better than NGAL/Cr (p = 0.033). CONCLUSIONS: The findings of the present study confirm that ACR and PCR are diagnostic biomarkers in T2D patients with decreased eGFR. NAPCR in these patients diagnostically only outperformed NGAL/Cr.

Potential Role and Excretion Level of Urinary Transferrin, KIM-1, RBP, MCP-1 and NGAL Markers in Diabetic Nephropathy.

BACKGROUND: Analyzing urinary biomarkers may provide better insight into pathophysiological mechanisms of diabetic kidney diseases. The study aimed to analyze the pattern of selected excreted urinary biomarkers and its correlation with albuminuria and estimated glomerular filtration rate (eGFR) in patients with type 2 diabetes. METHODS: A total of 185 patients with type 2 diabetes were categorized according to KDIGO guideline based on albuminuria and eGFR. The urinary markers (transferrin, KIM-1, RBP, MCP-1 and NGAL) were measured by ELISA. RESULTS: The urinary markers were associated with eGFR (total protein/Cr, p=0.001; RBP/Cr, p=0.007; MCP-1/Cr, p=0.023; NGAL/Cr, p=0.011) and albuminuria (total protein/Cr, p<0.001; transferrin, p<0.001; RBP/Cr, p<0.001; MCP-1/Cr, p<0.001; NGAL/Cr, p=0.002). CONCLUSION: The urinary marker levels (total protein, RBP, MCP-1, and NGAL) are elevated with severity of kidney damage and expressed more in progressive renal impairment.

Variation in the Level of Thyroid Markers in Association with Inflammation in Patients with Type 2 Diabetes.

BACKGROUND: A possible relationship between thyroid hormones and glucose metabolism in diabetes has already been established. OBJECTIVES: We aimed to evaluate the thyroid function markers and their relationship with inflammation, which is considered as a pathogenic condition of diabetes. METHODS: This cross-sectional study included 276 patients with type 2 diabetes. Serum levels of thyroid (TSH, FT4, and FT3) and inflammatory markers (CRP, IL-6, and TNF-α) were measured. RESULTS: The mean age of the subjects was 55.2 years and mean diabetes duration of 16.8 years. The inflammatory markers showed significant differences with the tertiles of TSH and thyroid hormones. TSH was significantly correlated with inflammatory markers, IL-6 (r = 0.13, P = 0.020) and TNF-α (r = 0.17, P = 0.003), while FT4 had a correlation only with TNF-α (r = 0.25, P = <0.001). FT3 was negatively correlated with inflammatory marker IL-6 (r = -0.14, P = 0.020), HbA1c (r = -0.12, P = 0 .040), and HOMA-IR (r = -0.17, P = 0.010). CONCLUSION: Abnormalities in the thyroid hormone metabolism are related to the increased inflammatory activity as well as insulin resistance, and are associated with the disorders of glucose metabolism.

Impact of Early Life or Intrauterine Factors and Socio-Economic Interaction on Diabetes - An Evidence on Thrifty Hypothesis.

Diabetes mellitus is one of the most concerning non-communicable diseases worldwide. The prevalence of diabetes increased rapidly by the influence of socioeconomic interactions. The thrifty hypothesis postulates that certain genes that are involved in positive selection promote efficient fat deposition and storage. This is beneficial for the survival of mankind in adverse conditions. However, in this modern society, these genes have become disadvantageous as people are significantly less likely to experience famines and nutrition shortages. The socioeconomic development that has occurred during the 20th century induced abundance of food supplies in almost all regions of the world. This has led to a rapid rise in the prevalence of obesity, and type 2 diabetes as a consequence. Boom of diabetic pandemic in newly developed countries compare with others those who developed gradually can be explain by thrifty hypothesis, as a result of the difference in the exposure to environmental factors and famine by the ancestors leads. The globalization, urbanization, lack of physical activity, intake of high calorie food and migration is major cause of pandemic emergence of diabetes in high as well as middle and low-income countries.

VCAM-1, ICAM-1 and selectins in gestational diabetes mellitus and the risk for vascular disorders.

Aim: Levels of VCAM-1, ICAM-1 and selectins in gestational diabetes mellitus (GDM) subjects are an indication of endothelial dysfunction predicting the future metabolic consequence via metabolic memory effect. Materials & methods: This cross-sectional study was conducted in 92 pregnant women and serum endothelial cell adhesion molecules were measured using Randox biochip analyzer. Results: Significantly elevated serum level of VCAM-1 was found in GDM subjects and in greater than equal to one parity categorized GDM group when compared with control. The correlation of parity and P-selectin was statistically significant in GDM subjects. Conclusion: Elevated levels of endothelial cell adhesion molecules in GDM women indicate an imbalance in vascular function. Transient hyperglycemia during pregnancy may induce persistent modifications to the memory cells and GDM subjects are more prone to develop future consequences.

Low magnesium level as an indicator of poor glycemic control in type 2 diabetic patients with complications.

AIM: Alteration in the metabolism of magnesium have an influence on different metabolic and signaling pathways involved in development of diabetes and its progression. Reduced magnesium level was associated with diabetes related complications. The aim of this study is to determine the serum levels of magnesium in diabetic patients having different complications and the association of magnesium with status of glycemic control. MATERIALS AND METHODS: This study was conducted among 88 type 2 diabetic patients, subdivided into two groups according to diabetic complications (with complications n = 55; without complications n = 33) and biochemical variables were measured. RESULTS: The serum magnesium level was decreased in diabetic patients having any complications (P = 0.039) or independent complication (nephropathy, P = 0.437; retinopathy, P = 0.038; neuropathy, P = 0.012 and macrovascular complication, P = 0.039), also decrease with increase in number of diabetic complications. Serum magnesium showed an inverse relation with glycemic parameters (HbA1c (r = -0.323; P = 0.002) and fasting blood glucose (r = - 0.321; P = 0.002)). CONCLUSION: The low levels of magnesium in diabetic complications, indicates the poor glycemic control in diabetic patients. Hence, maintaining the sufficient level of magnesium can control glycemia, thereby prevent the development of diabetic complications.

Association of urinary monocyte chemoattractant protein-1 (MCP-1) and kidney injury molecule-1 (KIM-1) with risk factors of diabetic kidney disease in type 2 diabetes patients.

PURPOSE: Urinary kidney injury molecule-1 and monocyte chemoattractant protein-1 are significance factors in the diagnosis and intervention of diabetic kidney diseases. This study determined levels of these proteins in diabetic patients with varying degrees of kidney disease and assessed their relationship with risk factors associated with diabetic kidney diseases. METHODS: A total of 185 patients with type 2 diabetes were divided into three groups [low risk (n = 47), moderate risk (n = 63), and high risk (n = 75)] based on the severity of diabetic kidney disease according to kidney disease: improving global outcomes guidelines. Both urinary kidney injury molecule-1 and monocyte chemoattractant protein-1 levels were measured by enzyme-linked immunosorbent assay. Student`s t test, analysis of variance, and Spearman's correlation were used for statistical analysis. RESULTS: The kidney injury molecule-1-to-creatinine ratio (P = 0.035) and monocyte chemoattractant protein-1-to-creatinine ratio (P < 0.001) increased significantly with the increase in kidney disease severity and varied according to different albuminuria statuses and estimated glomerular-filtration rates. The monocyte chemoattractant protein-1-to-creatinine ratio showed a significant correlation with hemoglobin A1c (P = 0.002) and inflammatory marker levels (interleukin-6, P = 0.005; tumor necrosis factor-α, P < 0.001). CONCLUSION: Urinary levels of both kidney injury molecule-1 and monocyte chemoattractant protein-1 represent distinguishing markers for the evaluation of diabetic kidney disease progression according to the associated degrees of albuminuria or/and the estimated glomerular-filtration rate. In addition, correlations between urinary monocyte chemoattractant protein-1 and glycemic and inflammatory marker levels revealed the role of hyperglycemia and chronic inflammation in the pathogenesis of diabetic kidney disease.

Potential role of serum fetuin-A in relation with pro-inflammatory, chemokine and adhesion molecules in diabetic kidney disease: a case-control study.

Inflammatory cytokine, adipokine and adhesion molecules are known to play a key role in pathogenesis of diabetic kidney disease (DKD). In this study, our aim was to investigate the role of fetuin-A in relation with pro-inflammatory cytokines (IL-6, IL-18), adipokines (adiponectin, leptin), chemokine (MCP-1), and adhesion molecules (ICAM-1, VCAM-1) in control and DKD subjects. We recruited a total of 224 type 2 diabetic (T2D) subjects. The control subjects were T2D with a normal albumin excrete (albumin-to-creatinine ratio-ACR ≤ 30 mg/g creatinine) and estimated glomerular filtration rate (eGFR) ≥ 60 (ml/min/1.73 m2), while cases were T2D subjects with albumin excrete (ACR ≥ 30 mg/g creatinine) and eGFR ≤ 60 (ml/min/1.73 m2). FBS, HbA1c, lipid profile (TC, LDL, HDL, triglyceride), ALT, AST, GGT, serum creatinine, BMI, blood pressure was evaluated in all the study subjects. Randox evidence biochip analyzer was used for measuring inflammatory cytokines, adipokines, and adhesion molecules by chemiluminescent assay. Serum fetuin-A and IL-18 were measured by ELISA kits. Serum fetuin-A levels were significantly decreased in DKD cases compare to control group [456.8 (299.2-649.0) µg/ml versus 670.6 (573.0-726.1) µg/ml; p < 0.001)]. Serum fetuin-A levels correlates significantly with IL-6, IL-18, TNF-α, PAI-1, leptin, resistin and ACR (p < 0.001). This study concludes that serum fetuin-A and pro-inflammatory markers (IL-18, IL-6, IL-1α and TNF-α) might play an important role in the pathophysiology and inflammatory process of DKD.

Molecular and Pathophysiological Mechanisms of Diabetic Retinopathy in Relation to Adhesion Molecules.

Diabetic Retinopathy (DR) is considered as a most common microvascular complication of diabetes affected by one in three people who are suffered for diabetes. Several pathophysiological mechanisms and adhesion molecules may play an etiologic role in the development of diabetes and its complications. The adhesion molecules located on both leucocytes and endothelial cells and considered as important molecules which can assessed the endothelial function. The functions of adhesion molecules involved in the cellular margination, slow rolling and transmigration of leukocytes. Hyperglycemia and its immediate biochemical sequelae or the low-grade inflammation directly alter endothelial function or influence endothelial cell functioning indirectly by induce oxidative stress and activates leukocytosis and leukocyte-endothelial cell interactions by the increased expression of adhesion molecules, growth factors, inflammatory factors, chemokines etc. and results DR. This review summarized the several pathophysiological mechanisms and role of adhesion molecules in disruption of homeostasis of vasculature by leukocytes in the development of diabetic retinopathy.

Angiopoietin-2 level as a tool for cardiovascular risk stratification in hypertensive type 2 diabetic subjects.

OBJECTIVES: This observational cross-sectional study aimed to investigate the relationship between serum Angiopoietin-2 (Ang-2) levels and cardiovascular (CVD) risk factors in drug controlled hypertensive diabetic subjects without cardiovascular complications. METHODS: All subjects were evaluated for fasting blood glucose (FBG), HbA1c, liver enzymes, lipid profile and serum Ang-2. RESULTS: Mean serum Ang-2 level was significantly higher in hypertensive diabetic subjects. In bivariate analysis in diabetic subjects with cardiovascular risk factors, Ang-2 positively correlated with waist circumference, body mass index (BMI), systolic blood pressure (SBP), FBG, HbA1c and triglycerides. In multivariate linear regression analysis, this association remained significant with FBG and triglycerides. Ang-2 levels were independently associated with CVD risk factors in drug controlled Type 2 diabetes (T2D) subjects. CONCLUSIONS: Further detailed studies in larger population with more attention is needed to consider Ang-2 level as a tool for CVD risk stratification in hypertensive diabetic subjects.

Variation in macro and trace elements in progression of type 2 diabetes.

Macro elements are the minerals of which the body needs more amounts and are more important than any other elements. Trace elements constitute a minute part of the living tissues and have various metabolic characteristics and functions. Trace elements participate in tissue and cellular and subcellular functions; these include immune regulation by humoral and cellular mechanisms, nerve conduction, muscle contractions, membrane potential regulations, and mitochondrial activity and enzyme reactions. The status of micronutrients such as iron and vanadium is higher in type 2 diabetes. The calcium, magnesium, sodium, chromium, cobalt, iodine, iron, selenium, manganese, and zinc seem to be low in type 2 diabetes while elements such as potassium and copper have no effect. In this review, we emphasized the status of macro and trace elements in type 2 diabetes and its advantages or disadvantages; this helps to understand the mechanism, progression, and prevention of type 2 diabetes due to the lack and deficiency of different macro and trace elements.