Background:Diabetic nephropathy (DN) is a major complication of Type 2 diabetes (T2D). Transforming growth factor-beta 1 (TGF-β1), a profibrotic cytokine, promotes fibrosis in DN, and its expression is affected by gene polymorphisms. This study aimed to determine the genotypic and allelic frequencies of the TGF-β1 gene (rs1800469 and rs1800470) as well as to test whether there is an association between TGF-β1 single-nucleotide polymorphisms (SNPs) and DN development in Saudi patients with T2D. Methods:This case-control study involved 204 samples, including 76 controls, 81 patients with T2D without DN, and 47 patients with T2D with DN. Genotyping was performed for two TGF-β1 SNPs using real-time PCR and TaqMan. Sanger sequencing was used for validation. Chi-square testing was conducted to confirm the strength of the association between genotype and allele frequencies and the risk of developing T2D and DN. Results:The distribution of rs1800469 and rs1800470 polymorphisms was not significantly different between the participants with T2D and control participants. No significant difference was observed in the same SNPs when comparing participants with DN and control participants. The albumin level in patients with DN with the A/A genotype was the highest, whereas it was the lowest in patients with the G/G genotype of TGF-β1 rs1800470. Conclusions:TGF-β1 rs1800470 and albuminuria are significantly associated with DN. TGF-β1 rs1800469 and rs1800470 were not significantly associated with T2D development and DN progression in Saudi patients with diabetes. Our findings suggest that continued investigation is necessary to understand the underlying mechanisms of TGF-β1 rs1800470 genotypes in DN pathogenesis.

Polymorphisms in antioxidant proteins, such as sirtuin 1 (SIRT1), may contribute to susceptibility to type 2 diabetes (T2D) and diabetic nephropathy (DN). SIRT1 rs3758391 and rs10823108 are associated with T2D and DN in other ethnicities; however, their role in susceptibility to T2D and DN in Saudi patients is unclear. Therefore, we investigated this association and examined the relationship between SIRT1 variants and serum SIRT1 protein levels. This case-control study included 126 patients with T2D (49 with DN and 77 controls). The associations between single-nucleotide polymorphisms and T2D with or without DN and SIRT1 expression were explored using logistic regression analysis. SIRT1 rs3758391 presence in controls and patients with T2D without DN of the T/C type and T/C genotype differed significantly. The presence of rs3758391 between patients with T2D with and without the T/C type and T/C genotype differed significantly. However, no association was observed with rs10823108. There was no association between rs3758391 and serum SIRT1 protein levels. SIRT1 rs3758391 was associated with T2D and DN in this Saudi cohort; however, rs10823108 showed no association. Serum SIRT1 was elevated in T2D irrespective of DN but was not correlated with rs3758391. These preliminary findings require further confirmation in future studies.

The etiology of women with gestational diabetes mellitus (GDM) and a greater risk of developing type 2 diabetes (T2D) after delivery remains unknown. This study aimed to investigate the global gene expression in four postpartum women with previous GDM (pGDM), three with T2D, and three with a history of normoglycemic pregnancy (controls). Total RNA was extracted from whole blood between March and May 2020. Global mRNA expression was determined using an Affymetrix Human Gene 2.0 ST Array. The expression of the selected focused genes was validated by RT-PCR. The microarray revealed 140 transcripts (p < 0.05, fold change cut-off ≥ 2) in patients with pGDM compared to controls. We identified 583 gene-altered transcripts between patients with T2D and controls. Interestingly, 60 transcripts had genes shared by pGDM or T2D versus the controls. The selected upregulated genes involved in inflammatory response, glycosylation, and death-like domains, according to the functional network analysis of pGDM (TNFAIP6, PDK3) and T2D (MMP9 and CARD6), showed similar trends to those obtained via microarray. Thus, these differentially expressed genes and their corresponding network and pathway analyses in women with pGDM and T2D offer valuable insights into the possible biological mechanisms of the progression of GDM to T2D.

BACKGROUND:Although oxidative stress has been implicated in PE, findings on antioxidant and oxidative DNA damage markers remain inconsistent. This study aimed to assess the plasma levels of the antioxidant biomarkers sirtuin 1 (SIRT1) and superoxide dismutase 2 (SOD2), as well as the oxidative DNA damage marker 8-hydroxy-2'-deoxyguanosine (8-OHdG), and to evaluate their correlation with clinical parameters in cases of PE. METHODS:This case-control study included 25 women aged 20-45 years, divided into two groups: the PE group (n = 14) and the control group (n = 11). Plasma SIRT1, SOD2, and 8-OHdG concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Pearson correlation coefficients were calculated to determine the relationships between plasma concentrations of these markers and specific clinical parameters in the PE group. RESULTS:Plasma levels of SIRT1 and SOD2 were significantly higher in the PE group compared to that in the control group, while no significant difference was observed in 8-OHdG. A significant positive correlation was observed between SIRT1 and both systolic and diastolic blood pressures in the PE group compared to that in the control group. In addition, a borderline significant positive association was observed between SOD2 and diastolic blood pressure in the PE group compared to that in the control group. CONCLUSION:Increased levels of both plasma SIRT1 and SOD2, along with their positive correlation with blood pressure, suggest a potential role for oxidative stress and antioxidant response in the pathogenesis of PE.

Aims: Fetal exposure to hyperglycemia in utero have been suggested to induce epigenetic changes through expression of various miRNAs, and cause dysfunctional endothelium connected with elevated risk of cardiovascular disease (CVD) in offspring during early adulthood. In this study, we investigated whether hyperglycemia-induced changes in the expression of 28 fetal endothelial microRNAs (miRNAs) are associated with endothelial dysfunction and CVD. Methods: Differentially expressed miRNAs in TaqMan miRNA human arrays were quantified using qPCR. Results: The expression of miR-140-3p, miR-1307-5p, miR-342-3p, and miR-16-5p was significantly reduced in human umbilical vein endothelial cells (HUVECs) from females with gestational diabetes (GDM-UVECs) compared with that of the control group. Meanwhile, in type 2 diabetes (T2D-HUVECs), miR-126-3p and miR27a-5p were significantly reduced, while miR-27b-3p was significantly increased. Furthermore, miR-29b-3p expression was upregulated in both GDM- and T2D-HUVECs compared with that in the control. The expression of fibroblast growth factor 11 (FGF11) mRNA-a target of dysregulated miR-342-3p-was downregulated in GDMHUVECs compared with that in the control. Conclusions: Altered miRNA and target FGF11 mRNA expression may contribute mechanistically to endothelial dysfunction in diabetic HUVECs when maintained under normal glucose conditions for several passages.

Obesity is a major global health issue associated with an increased risk of early-onset metabolic disorders and chronic inflammation. Identifying the epigenetic mechanisms that contribute to obesity-related metabolic and inflammatory dysregulation is crucial for developing effective prevention and treatment strategies. This pilot study aimed to investigate the effects of obesity on the expression of microRNAs (miRNAs) related to metabolic disorders in human peripheral blood mononuclear cells from metabolically healthy obese subjects and non-obese controls. Differentially expressed miRNAs in TaqMan human miRNA arrays were quantified using quantitative PCR. To validate the robustness and generalizability of our findings, we performed cross-validation using the publicly available GSE155096 dataset. The expression of miR-145-5p was significantly increased (4.913-fold change) in obese individuals compared to the non-obese control group. Two miRNAs, miR-27b-3p and miR-17-5p, were downregulated 2.207- and 1.448-fold, respectively, approaching significance. A positive correlation was established between miR-145-5p and free triiodothyronine, eosinophils, and vitamin D. A cross-validation analysis confirmed the direction of change for these key miRNAs. The data suggest that miR-145-5p, miR-27b-3p, and miR-17-5p could be implicated in the progression of obesity in causing metabolic abnormalities, clarifying how molecular factors cause the metabolic deregulation associated with obesity.

ObjectivesThis case-control study explored the association of sirtuin 1 polymorphisms (rs10997868 and rs730821) with metabolic parameters, smoking duration, sirtuin 1 levels, and the oxidative stress biomarker 8-hydroxy-2'-deoxyguanosine in healthy smokers in Saudi Arabia.MethodsGenomic DNA was isolated from peripheral blood samples, and rs10997868 and rs730821 polymorphism genotyping was performed for 43 healthy smokers and 33 healthy controls using real-time polymerase chain reaction. Plasma sirtuin 1 and 8-hydroxy-2'-deoxyguanosine levels were measured using enzyme-linked immunosorbent assay.ResultsSirtuin 1 levels were significantly higher in healthy smokers than in controls, while 8-hydroxy-2'-deoxyguanosine levels were not significantly lower in healthy smokers. There was no significant association of sirtuin 1 protein expression and 8-hydroxy-2'-deoxyguanosine levels with rs10997868 and rs730821. Our findings showed a significant association between smoking duration and rs10997868 single nucleotide polymorphism, which was highest in those with the C/C genotype and lowest in those with the A/C genotype (p = 0.035). In addition, the serum cholesterol level was highest in the C/C group and lowest in the A/C group (p = 0.029). None of the studied factors (smoking duration, serum cholesterol, fasting blood sugar, or serum triglycerides) was significantly associated with sirtuin 1 rs730821.ConclusionsOur study showed that sirtuin 1 upregulation and polymorphisms are associated with smoking duration and lipid levels in healthy smokers.

Objective: To assess the changes in the expression of anti-oxidant and pro-oxidant enzymes in patients with type 2 diabetes and diabetic nephropathy, and to examine their correlation with insulin resistance. Method: The case-control study was conducted from March to November 2021 at the King Abdulaziz University Hospital, Jeddah, Saudi Arabia, and comprised adult patients of either gender with diabetic nephropathy in DN group, patients with type 2 diabetes but without diabetic nephropathy in T2D group, and non-diabetic individuals in the control group. Serum insulin levels were measured using a modular analyser, while nicotinamide adenine dinucleotide phosphate oxidase, glutathione S-transferase and superoxide dismutase 3 levels were measured using enzyme-linked immunosorbent assay. Data was analysed using SPSS 29.0.1. Results: Of the 74 subjects, 45(60.8%) were females and 29(39.2%) were males. The overall mean age was 53±14 years. There were 20(27%) patients in DN group with mean age 60±11 years, 29(39.2%) patients in T2D group with mean age 56±12 years, and 25(33.8%) in the control group with mean age 43±11 years. Nicotinamide adenine dinucleotide phosphate oxidase levels were significantly lower in T2D and DN groups than in the control group (p<0.05). Glutathione S-transferase levels were significantly lower in the DN group than in the control group (p<0.05). T2D and DN groups had significantly lower superoxide dismutase 3 levels than the control group (p<0.05). Glutathione S-transferase levels correlated positively with glycated haemoglobin levels in the DN group, and negatively with fasting blood glucose levels in the T2D group (p<0.05). Superoxide dismutase 3 levels were negatively correlated with insulin and homeostatic model assessment of insulin in T2D group (p<0.05). Conclusion: Type 2 diabetes and diabetic nephropathy induced changes in the levels of superoxide dismutase 3, glutathione S-transferase and nicotinamide adenine dinucleotide phosphate oxidase. Low levels of superoxide dismutase 3 in type 2 diabetes correlated with insulin resistance, suggesting the need for anti-oxidant replacement therapy as part of diabetes control measures to prevent diabetic nephropathy. Key Words: Type 2 diabetes mellitus, Diabetic nephropathy, Insulin resistance, Oxidative stress, Anti-oxidant.

Oxidative stress has been hypothesized to play a crucial role in the complications of type 2 diabetes (T2D). Hyperglycaemia-mediated increases in free radicals have a deleterious effect on cellular compartments and nucleic acids, leading to imbalances between free radicals and antioxidant enzymes. This case–control study comprised two groups with 100 participants (50 T2D and 50 DN patients) and aimed to examine the association between single-nucleotide polymorphisms (SNPs) in the genes encoding nicotinamide adenine dinucleotide phosphate oxidase (NOX), glutathione S-transferase P (GSTP1), and glutathione peroxidase 1(GPX1) and diabetic nephropathy (DN) risk in patients with T2D. An SNP genotyping assay was performed using TaqMan assay and real-time PCR to identify the SNPs (NOX rs4673, GSTP rs1695, GPX1 rs1050450). The Sanger method was used to validate our findings. Fisher chi-square analyses revealed no significant differences in these genes when comparing T2D patients with and without DN. Our findings suggest no association between the rs4673, rs1695, and rs1050450 SNPs and DN in Saudi patients with T2D.

From an operational, administrative and scientific perspective, cyber security is one of the most important factors affecting the quality of the educational process and the continuity of operations in universities.Therefore, it is crucial to take the necessary security measures to ensure the security of cloud computing.The aim of this study was to identify the key cybersecurity issues in cloud computing and clarify the cybersecurity regulations for cloud computing in the Kingdom of Saudi Arabia.In addition, the study comprehensively examined the current state of cybersecurity for cloud computing at Taibah University.The study used a descriptive methodology that included analytical aspects and a case study, and used a checklist based on cybersecurity controls for cloud computing issued by the National Cybersecurity Authority as a data collection tool.The data were supplemented by content analysis of primary sources from the cybersecurity department of the Taibah University website and openended interviews with a sample of participants consisting of cybersecurity engineers and IT engineers from Taibah University.The study found several findings, including the need to implement security controls and standards to ensure the security of data and content stored in the cloud.It

INTRODUCTION:Despite strong and consistent epidemiological evidence linking cigarette smoking to several cardiovascular diseases (CVDs), the association between smoking intensity and CVD risk factors remains unclear. This study aimed to explore the possible effects of cigarette smoking on cardiometabolic risk in healthy individuals. METHODS:This cross-sectional study was conducted between November 2022 and June 2023. Consecutive sampling was performed to include 160 healthy participants: 100 smokers with 60 males and 40 females; and 60 age- and sex-matched non-smokers with 36 males and 24 females. Blood samples were taken from each participant to assess their cardiometabolic function: lipid profile, von Willebrand factor (vWF), high-sensitivity cardiac troponin I (hs-cTnI), and fibrinogen levels; and liver function using an automated enzymatic method. In addition, blood sugar level, body mass index (BMI), and blood pressure were recorded. RESULTS:Smokers had significantly higher vWF functional activity and hs-cTnI but significantly lower albumin and total bilirubin levels than non-smokers (65.87 ± 19.07 vs 56.45 ± 6.59, respectively, p<0.001; 0.0382 ± 0.0077 vs 0.0147 ± 0.0105, respectively, p<0.001; and 4.63 ± 0.32 vs 4.74 ± 0.28, respectively, p=0.026). The number of cigarettes consumed daily was associated positively and significantly with plasma levels of low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, vWF functional activity, and hs-cTnI but were negatively associated with total bilirubin. Moreover, heavy smokers had a significantly higher BMI and waist-to-hip ratio among male smokers than non-smokers. CONCLUSIONS:Cigarette smoking was associated with increased dyslipidemia, BMI, and central obesity, in addition to higher vWF functional activity. Altogether, increased hs-cTnI levels in smokers indicate a higher susceptibility to CVD.

Foetuses exposed to maternal gestational diabetes (GDM) and type 2 diabetes (T2D) have an increased risk of adverse perinatal outcomes. Epigenetic mechanisms, including DNA methylation and histone modifications, may act as mediators of persistent metabolic memory in endothelial cells (ECs) exposed to hyperglycaemia, even after glucose normalization. Therefore, we investigated alterations in global DNA methylation and epigenetic modifier expression (DNMT1, DNMT3a, DNMT3b, HDAC1, and HDAC2) in human umbilical vein ECs (HUVECs) from the umbilical cords of mothers with GDM (n = 8) and T2D (n = 3) compared to that of healthy mothers (n = 6). Global DNA alteration was measured using a 5-methylation cytosine colorimetric assay, followed by quantitative real-time polymerase chain reaction to measure DNA methyltransferase and histone acetylase transcript expression. We revealed that DNA hypermethylation occurs in both GDM- and T2D-HUVECs compared to that in Control-HUVECs. Furthermore, there was a significant increase in HDAC2 mRNA levels in GDM-HUVECs and increase in DNMT3b mRNA levels in T2D-HUVECs. Overall, our results suggest that GDM and T2D are associated with global DNA hypermethylation in foetal endothelial cells under normoglycemic conditions and the aberrant mRNA expression of HDAC2 and DNMT3b could play a role in this dysregulation.

Aim of the study: To explore the effects of cigarette smoking on cardiometabolic risk in Saudis in Jeddah City. Methods: Following a descriptive case-control approach, the study included 160 healthy Saudi adults who were sampled consecutively (100 smokers [60 males and 40 females] and 60 non-smokers [36 males and 24 females]). Personal characteristics, smoking patterns, and cardiometabolic risk assessment were included in a questionnaire. Each participant’s cardiometabolic functions were assessed by drawing blood. Results: Among participant smokers, 80% smoked one pack daily, while 20% smoked more than one pack daily. Smokers had insignificantly higher levels of total cholesterol, LDL-cholesterol, and non-HDL, significantly higher vWF functional activity, and high-sensitivity cardiac troponin I (p<0.001 for both), but significantly lower albumin and total bilirubin levels than non-smokers (p=0.026, and p<0.001, respectively). The number of daily cigarettes consumed correlated positively and significantly with plasma levels of LDL-cholesterol (r=0.225, p=0.004), non-HDL cholesterol (r=0.220, p=0.005), vWF function activity (r=410, p<0.001), high-sensitivity cardiac troponin I (r=0.686, p<0.001), but negatively correlated with total bilirubin (r=-0.459, p<0.001). Moreover, the cigarette smoking intensity correlated positively and significantly with the participant’s systolic blood pressure (r=0.303, p<0.001) and diastolic blood pressure (r=0.300, p<0.001), body mass index (r=0.448, p<0.001), and waist-to-hip ratio (r=0.493, p<0.001). Those who had smoked for more than 10 years had significantly higher plasma levels of triglycerides (p=0.031), total cholesterol (p=0.023), LDL cholesterol (p=0.011), non-HDL cholesterol (p=0.008), vWF functional activity (p<0.001), systolic blood pressure (p=0.011), and diastolic blood pressure (p=0.023). Moreover, heavy smokers had a significantly higher BMI (p=0.001) and waist-to-hip ratio among male smokers (p=0.003). Conclusions: Cigarette smoking is associated with increased dyslipidemia, body mass index, and central obesity, in addition to higher vWF functional activity. Increased hs-cTnI levels in smokers indicate a higher risk of heart failure and cardiovascular death. Keywords: smoking, cardiometabolic risk, lipid profile, liver function, troponin.

Abstract Background This study aimed to explore the possible effects of cigarette smoking on the cardiometabolic risk of apparently healthy Saudi individuals in Jeddah City.Methods A blood sample was withdrawn from each participant (100 smokers [60 males and 40 females] and 60 nonsmokers [36 males and 24 females]) for the assessment of their cardiometabolic functions (lipid profile, von Willebrand factor (vWF), Troponin I) and liver function tests using an automated enzymatic method.Results Among participant smokers, 80% smoked one pack daily, while 20% smoked more than one pack daily. Smokers had significantly higher vWF functional activity and high-sensitivity cardiac troponin I (p < 0.001 for both), but significantly lower albumin and total bilirubin levels than nonsmokers (p = 0.026, and p < 0.001, respectively). The number of daily cigarettes consumed correlated positively and significantly with plasma levels of LDL-cholesterol (r = 0.225, p = 0.004), non-HDL cholesterol (r = 0.220, p = 0.005), vWF function activity (r = 410, p < 0.001), high-sensitivity cardiac troponin I (r = 0.686, p < 0.001), but negatively correlated with total bilirubin (r=-0.459, p < 0.001). Moreover, heavy smokers had a significantly higher BMI (p = 0.001) and waist-to-hip ratio among male smokers compared to control non- smokers (p = 0.003). Thus,Conclusions Cigarette smoking is associated with increased dyslipidemia, body mass index, and central obesity, in addition to higher vWF functional activity. Increased hs-cTnI levels in smokers, indicating a higher susceptibility to heart failure and cardiovascular mortality among smokers.

Abstract Introduction One of the complications of diabetes mellitus (DM) is diabetic nephropathy (DN), which plays a significant role in the progression of end‐stage renal disease. Oxidative stress is implicated in DN pathogenesis, and genetic variations in antioxidant enzymes such as superoxide dismutase 2 (SOD2) and catalase (CAT) may contribute to the susceptibility. This study aimed to investigate the potential association between single nucleotide polymorphisms (SNPs) in antioxidant enzymes, specifically SOD2 rs4880 and CAT rs769217, and the risk of T2D and susceptibility to DN within the Saudi population. Methods This case–control study included 150 participants, comprising 50 patients with T2D without DN (group 1), 50 patients with T2D with DN (group 2), and 50 healthy participants (group 3). The samples were genotyped using real‐time PCR for SOD2 rs4880 and CAT rs769217 SNPs. Sanger sequencing was used for validation. Statistical analyses were performed to explore associations between these SNPs and T2D with or without DN. Results No significant difference was observed in CAT rs769217 expression between the groups. However, a significant difference was observed in SOD2 rs4880 expression between the healthy controls and patients with T2D with DN (p = .028). Furthermore, SOD2 rs4880 was associated with approximately threefold increased risk of DN in patients with T2D compared to that in healthy participants (odds ratio [OR] = 2.99 [1.31–6.83]). Validation through Sanger sequencing further confirmed these findings. Conclusions The findings of this study provide evidence that SOD2 rs4880 SNP may contribute to inadequate defence by the antioxidant enzyme, SOD2, against DM‐induced oxidative stress and thus cause DN in Saudi patients with T2D. Therefore, SOD2 rs4880 may serve as a predictive marker to prevent the development and progression of DN in patients with T2D.

Stem cells are a versatile source for cell therapy. Their use is particularly significant for the treatment of neurological disorders for which no definitive conventional medical treatment is available. Neurological disorders are of diverse etiology and pathogenesis. Alzheimer’s disease (AD) is caused by abnormal protein deposits, leading to progressive dementia. Parkinson’s disease (PD) is due to the specific degeneration of the dopaminergic neurons causing motor and sensory impairment. Huntington’s disease (HD) includes a transmittable gene mutation, and any treatment should involve gene modulation of the transplanted cells. Multiple sclerosis (MS) is an autoimmune disorder affecting multiple neurons sporadically but induces progressive neuronal dysfunction. Amyotrophic lateral sclerosis (ALS) impacts upper and lower motor neurons, leading to progressive muscle degeneration. This shows the need to try to tailor different types of cells to repair the specific defect characteristic of each disease. In recent years, several types of stem cells were used in different animal models, including transgenic animals of various neurologic disorders. Based on some of the successful animal studies, some clinical trials were designed and approved. Some studies were successful, others were terminated and, still, a few are ongoing. In this manuscript, we aim to review the current information on both the experimental and clinical trials of stem cell therapy in neurological disorders of various disease mechanisms. The different types of cells used, their mode of transplantation and the molecular and physiologic effects are discussed. Recommendations for future use and hopes are highlighted.

OBJECTIVES:This study reports the levels of cytokines, chemokines, and growth factors previously identified as taking part in the pathology of atherosclerosis in human umbilical vein endothelial cells derived from mothers with type 2 diabetes and compares them with those in human umbilical vein endothelial cells derived from healthy mothers under normal glucose conditions.METHODS:Cytokine analysis measures of human umbilical vein endothelial cell lysates were obtained using a multiple analyte profiling (xMAP) assay based on magnetic bead-based technology, using the MAGPIX instrument. The correlation between cytokines, chemokines, and growth factors was examined statistically in human umbilical vein endothelial cells derived from mothers with type 2 diabetes.RESULTS:This study showed that the expression of proinflammatory cytokine interleukin-1 alpha was significantly greater in human umbilical vein endothelial cells derived from mothers with type 2 diabetes than those derived from healthy mothers. The protein level of granulocyte colony-stimulating factor was higher in human umbilical vein endothelial cells derived from mothers with type 2 diabetes than those derived from healthy mothers. A significant positive correlation was demonstrated between the protein expression of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor in human umbilical vein endothelial cells derived from mothers with type 2 diabetes.CONCLUSION:Diabetes evokes a persistent inflammatory phenotype in human umbilical vein endothelial cells, as indicated by the enhanced production of cytokines and growth factors under normal glucose conditions.

Gestational diabetes mellitus (GDM) affects one in four Saudi women and is associated with high risks of cardiovascular diseases in both the mother and foetus. It is believed that endothelial cells (ECs) dysfunction initiates these diabetic complications. In this study, differences in the transcriptome profiles, cell cycle distribution, and mitochondrial superoxide (MTS) between human umbilical vein endothelial cells (HUVECs) from GDM patients and those from healthy (control) subjects were analysed. Transcriptome profiles were generated using high-density expression microarray. The selected four altered genes were validated using qRT-PCR. MTS and cell cycle were analysed by flow cytometry. A total of 84 altered genes were identified, comprising 52 upregulated and 32 downregulated genes in GDM.HUVECs. Our selection of the four interested altered genes (TGFB2, KITLG, NEK7, and IGFBP5) was based on the functional network analysis, which revealed that these altered genes are belonging to the highest enrichment score associated with cellular function and proliferation; all of which may contribute to ECs dysfunction. The cell cycle revealed an increased percentage of cells in the G2/M phase in GDM.HUVECs, indicating cell cycle arrest. In addition, we found that GDM.HUVECs had increased MTS generation. In conclusion, GDM induces persistent impairment of the biological functions of foetal ECs, as evidenced by analyses of transcriptome profiles, cell cycle, and MTS even after ECs culture in vitro for several passages under normal glucose conditions.

BACKGROUND:Allicin (ACN), a sulfoxide in freshly crushed garlic, is known for its diverse bioactive properties. Among the most notable effects of ACN is its antitumor activity against a wide array of cancer types. Thus, ACN may be a promising anticancer therapeutic. Nevertheless, chemotherapy-induced anemia is a major obstacle in cancer management with a prevalence of up to 70%. Although the pathophysiology behind it remains elusive, a number of medications known to cause anemia in patients have been shown to induce premature programmed cell death in red blood cells (RBCs) known as eryptosis. This study, thus, investigates the anticancer potential of ACN against THP-1 monocytic leukemia cells, its toxic effects on human RBCs, and delineate the underlying biochemical mechanisms.METHODS:Cytotoxicity was detected using the MTT assay, while hemoglobin leakage was used as a surrogate for hemolysis which was photometrically measured. Major eryptotic events were examined using flow cytometry with fluorescent probes. Phosphatidylserine (PS) exposure was detected by Annexin-V-FITC, cytosolic calcium with Fluo4/AM, and reactive oxygen species with H2DCFDA.RESULTS:Our results show that ACN induces hemolysis in a dose-dependent fashion, which is significantly abrogated in absence of extracellular calcium. Moreover, ACN stimulates PS exposure, intracellular calcium overload, and oxidative stress. Using small-molecule inhibitors, we demonstrate that the pro-eryptotic activity of ACN is ameliorated in presence of zVAD(OMe)-FMK, SB203580, and D4476.CONCLUSION:ACN possesses both hemolytic and eryptotic properties mediated through elevated intracellular calcium levels, oxidative stress, caspase, p38 MAPK, and CK1α.

Hyperglycemia can induce tissue damage, by increasing the oxidative stress and pro-inflammatory mediators, whereas the secretome of stem cells helps in the restoration of tissue homeostasis. Resveratrol (RSV), a polyphenolic compound, is reported to exert both anti-inflammatory and antioxidant effects on both in vitro and in vivo models of health and disease. In this study, we evaluated the effects of high glucose (HG), at either 28 mM, and 41 mM, RSV (17 mu M), or in combination, on the cell morphology, proliferation, and regulation of cytokines in human Wharton's jelly-derived stem cells (hWJSCs) in vitro. Treatment with HG reduced the hWJSCs proliferation compared with the control but did not affect the cell morphology. In addition, exposure to RSV alone or in combination with HG significantly enhanced cell proliferation. The cytokine analysis of the supernatants of the hWJSCs by Multiple Analyte Profiling (xMAP) based on magnetic bead-based technology using MAGPIX instrument showed that the expression of pro-inflammatory cytokine interleukin-6 (IL-6) was significantly reduced in all the treatment groups, while the interleukin-8 (IL-8) or chemokine (C-X-C motif) ligand 8 (CXCL8) was reduced upon treatment with RSV or RSV with HG at 72 h. Meanwhile, the granulocyte-macrophage colony-stimulating factor (GM-CSF) secretion, was differentially regulated in the treatment groups when compared with control. We, therefore, concluded that HG inhibited the proliferation and caused cellular damage in hWJSCs. However, the RSV treatment helped to reverse these detrimental effects possibly by the differential regulation of IL-6, IL-8, and GM-CSF in hWJSCs.