Nitric Oxide Synthase Gene Research Articles

Iron oxide polyaniline-coated nanoparticles modulate tumor microenvironment in breast cancer: an in vitro study on the reprogramming of tumor-associated macrophages

BackgroundBreast cancer is the neoplastic disease with the highest incidence and mortality in the female population worldwide. Treatment remains challenging due to various factors. Therefore, it is of great importance to develop new therapeutic strategies that promote the safe destruction of neoplastic cells without compromising patients' quality of life. Among advances in the treatment of breast cancer, immunotherapy stands out as a promising trend. Recent studies have demonstrated the potential of iron oxide nanoparticles in promoting the reprogramming of M2 macrophages (pro-tumor phenotype) into M1 macrophages (anti-tumor phenotype) within the tumor microenvironment, resulting in potent antitumor effects. In this study, the effect of polyaniline-coated iron oxide nanoparticles (Pani/y-Fe2O3) on macrophage polarization and breast cancer cell death was investigated.MethodsThe non-cytotoxic concentration of nanoparticles was determined using the MTT assay. For in vitro co-culture experiments, breast cancer cell lines MCF -7 and MDA-MB -231 and macrophages THP-1 were co-cultured in a Transwell system and then the effects of Pani/y-Fe2O3 on cell viability, gene expression, cytokine profile, and oxidative stress markers were investigated.ResultsThe results showed that Pani/y-Fe2O3 nanoparticles induced M2-to-M1 macrophage polarization in both cell lines through different pathways. In MCF -7 and THP-1 macrophage co-culture, the study showed a decrease in cytokine levels IL -1β, upregulation of M1-associated genes (IL-12, TNF-α) in macrophages, resulting in increased MCF -7 cell death by apoptosis (caspase 3/7+). In MDA-MB -231 co-cultures, increases in cytokines IL -6, IL -1β, and oxidative stress markers were observed, as well as upregulation of the inducible nitric oxide synthase (iNOS) gene in macrophages, leading to tumor cell death via apoptosis-independent pathways (Sytox+).ConclusionsThese findings highlight the potential of Pani/y-Fe2O3 as a promising therapeutic approach in the context of breast cancer treatment by effectively reprogramming M2 macrophages into an anti-tumor M1 phenotype, Pani/y-Fe2O3 nanoparticles demonstrated the ability to elicit antitumor effects in both MCF-7 and MDA-MB-231 breast cancer cell lines.

Glucomannan enhanced the macrophage activity in exposure to methicillin-resistant Staphylococcus aureus (MRSA): in-vitro study.

The increasing number of methicillin-resistant Staphylococcus aureus persuade the need for preventive measures. Glucomannan is a polysaccharide choice for developing immunological strategies. This study aimed to investigate changes in gene expression and phagocytic activity of macrophage cells in the presence of glucomannan. The effect of different concentrations of glucomannan (25, 50, and 100 μg/mL) on the phagocytic activity of macrophage cells was measured using the colony count method. The expression of Tumor Necrosis Factor-alpha (TNF-α) and Inducible Nitric Oxide Synthase (iNOS) genes was evaluated by Real-Time PCR. The concentrations of glucomannan significantly reduced the bacterial Colony-Forming Unit (CFU) and increased the phagocytic activity of macrophage cells. The maximum effect of glucomannan on iNOS and TNF-A genes expression was 100 μg/mL. Glucomannan should be considered an adjuvant that stimulates the immune system. It may increase the expression of TNF-α and iNOS genes and the phagocytic activity of macrophage cells against methicillin-resistant Staphylococcus aureus.

Endothelial Nitric Oxide Synthase T-786C and G-894T Gene Polymorphisms: A Risk Assessment of Coronary Heart Disease.

The polymorphism of the endothelial nitric oxide synthase (eNOS) gene is thought to enhance the risk of coronary heart disease (CHD). The most protruding reason for endothelial dysfunction is the diminished production of NO. In the eNOS encoding gene, a number of mutations were associated with decreased NO effect promoting the presence of CHD. This study aims to look at the role of polymorphisms in the eNOS genes G-894T and T-786C in young South Indians. From January 2022 to May 2022, 91 angiographically proven CHD subjects attending the Department of Cardiology and Medicine and 91 controls from a master health checkup in the age group of 45 years participated in this observational cross-sectional study at SRM Medical College Hospital and Research Centre in Chennai, Tamil Nadu, India. Overnight fasting plasma samples were taken for analysis of the lipid profile as well as NO by Griess reaction utilizing an enzyme-linked immunosorbent assay (ELISA) technique. Polymerase chain reaction (PCR) and restricted fragment length polymorphism (RFLP) were used to amplify the T-786C and G-894T eNOS genes. When compared to controls, the mean level of serum NO in CHD patients was considerably lower. For eNOS T-786C polymorphism, the distribution of TC genotype (p = 0.017), odds (OD) ratio = 2.1, CC genotype (p = 0.011), OD ratio = 3.75, and minor C allele frequency (p = 0.001). And for eNOS G-894T polymorphism, the distribution of GT genotype (p = 0.01), TT genotype (p =0.02) with OD ratio and minor T allele frequency (p = 0.002) with OD ratio = 1.07 and 0.83. Our study concludes that the polymorphism of eNOS T-786C and G-894T genes may be the main causative association for the presence of CHD. How to cite this article: Jaishankar T, Shivasekar M, Vinodhini VM. Endothelial Nitric Oxide Synthase T-786C and G-894T Gene Polymorphisms: A Risk Assessment of Coronary Heart Disease. J Assoc Physicians India 2023;71(9):45-50.

Attenuation of Oxidative Damage via Upregulating Nrf2/HO-1 Signaling Pathway by Protease SH21 with Exerting Anti-Inflammatory and Anticancer Properties In Vitro.

Oxidative damage and inflammation are among the very significant aspects interrelated with cancer and other degenerative diseases. In this study, we investigated the biological activities of a 25 kDa protease (SH21) that was purified from Bacillus siamensis. SH21 exhibited very powerful antioxidant and reactive oxygen species (ROS) generation inhibition activity in a dose-dependent approach. The mRNA and protein levels of antioxidant enzymes such as superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (GPx-1) were enhanced in the SH21-treated sample. SH21 also increased the transcriptional and translational activities of NF-E2-related factor 2 (Nrf2) with the subsequent development of detoxifying enzyme heme oxygenase-1 (HO-1). In addition, SH21 showed potential anti-inflammatory activity via inhibition of nitric oxide (NO) and proinflammatory cytokines, such as TNF-α, IL-6, and IL-1β, production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. At concentrations of 60, 80, and 100 μg/mL, SH21 potentially suppressed nitric oxide synthase (iNOS) and cytokine gene expressions. Furthermore, SH21 significantly released lactate dehydrogenase (LDH) enzyme in cancer cell supernatant in a concentration-dependent manner and showed strong activity against three tested cancer cell lines, including HL-60, A549, and Hela. Our results suggest that SH21 has effective antioxidant, anti-inflammatory, and anticancer effects and could be an excellent therapeutic agent against inflammation-related diseases.

Prospective validation of VEGF and eNOS polymorphisms as predictors of first-line bevacizumab efficacy in patients with metastatic colorectal cancer

Bevacizumab (Bev) plus chemotherapy is a standard first-line treatment in metastatic colorectal cancer (mCRC), however to date no predictive factors of response have been identified. Results of our previous analysis on patients enrolled in a randomized prospective phase III multicenter study (ITACa study) showed a predictive value of Vascular Endothelial Growth Factor (VEGF) polymorphism (VEGF + 936), a 27-nucleotide variable number tandem repeat (VNTR) of the endothelial nitric oxide synthase (eNOS) gene and eNOS + 894 polymorphism. mCRC patients, treated with Bev plus chemotherapy, were included in this prospective validation trial. eNOS + 894G > T was analyzed by Real time PCR, while the eNOS VNTR and VEGF + 936C > T were determined by standard PCR and direct sequencing analysis. These polymorphisms were assessed in relation to progression-free survival (PFS), overall survival (OS) and objective response rate (ORR). These three polymorphisms were not predictive of PFS (p 0.91, 0.59 and 0.09, respectively), and OS (p 0.95, 0.32 and 0.46, respectively). Moreover, the haplotype analyses did not confirm what was found in our previous study; patients bearing a specific haplotype of eNOS had not significantly improved outcomes. This prospective study failed to validate the predictive impact of eNOS and VEGF polymorphisms for response to Bev plus first-line chemotherapy in mCRC patients.

A novel imidazo[1,2-a]pyridine derivative and its co-administration with curcumin exert anti-inflammatory effects by modulating the STAT3/NF-κB/iNOS/COX-2 signaling pathway in breast and ovarian cancer cell lines.

Imidazo[1,2-a]pyridine derivatives with diverse pharmacological properties and curcumin, as a potential natural anti-inflammatory compound, are promising compounds for cancer treatment. This study aimed to synthesize a novel imidazo[1,2-a]pyridine derivative, (MIA), and evaluate its anti-inflammatory activity and effects on nuclear factor-κB (NF-κB) and signal transducer and activator of transcription 3 (STAT3) pathways, and their target genes, alone and in combination with curcumin, in MDA-MB-231 and SKOV3 cell lines. We evaluated the interaction between imidazo[1,2-a]pyridine ligand, curcumin, and NF-κB p50 protein, using molecular docking studies. MTT assay was used to investigate the impacts of compounds on cell viability. To evaluate the NF-κB DNA binding activity and the level of inflammatory cytokines in response to the compounds, ELISA-based methods were performed. In addition, quantitative polymerase chain reaction (qPCR) and western blotting were carried out to analyze the expression of genes and investigate NF-κB and STAT3 signaling pathways. Molecular docking studies showed that MIA docked into the NF-κB p50 subunit, and curcumin augmented its binding. The MTT assay results indicated that MIA and its combination with curcumin reduced cell viability. According to the results of the ELISA-based methods, MIA lowered the levels of inflammatory cytokines and suppressed NF-κB activity. In addition, real-time PCR and Griess test results showed that the expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) genes, and nitrite production were reduced by MIA. Furthermore, the western blotting analysis demonstrated that MIA increased the expression of inhibitory κB (IκBα) and B-cell lymphoma 2 (Bcl-2)-associated X proteins (BAX), and suppressed the STAT3 phosphorylation, and Bcl-2 expression. Our findings revealed that curcumin had a potentiating role and enhanced all the anti-inflammatory effects of MIA. This study indicated that the anti-inflammatory activity of MIA is exerted by suppressing the NF-κB and STAT3 signaling pathways in MDA-MB-231 and SKOV3 cancer cell lines.

Triterpenoid and Coumarin Isolated from Astilbe grandis with Anti-Inflammatory Effects through Inhibiting the NF-κB Pathway in LPS-Induced RAW264.7 Cells.

The roots of Astilbe grandis, known as "Ma sang gou bang", are used as a Miao traditional medicine with anti-inflammatory and analgesic properties. However, the active components and mechanism of action of this plant remain mostly uncharacterized. The aim of this study was to identify its active components and verify their pharmacological activity. The extract of A. grandis root was separated using various chromatographic methods. As a result, we obtained one novel triterpenoid, named astigranlactone (1), which has an unusual lactone moiety formed between C-7 and C-27. Additionally, a known coumarin compound, 11-O-galloyl bergenin (2) was isolated from this plant. The structures of these two compounds were elucidated by extensive NMR experiments in conjunction with HR-ESI-MS data. To the best of our knowledge, both compounds were isolated from this species for the first time. Moreover, we tested the anti-inflammation effect of the two compounds by establishing a cellular inflammation model induced by LPS in RAW264.7 cells. The effect of different concentrations of these compounds on the activity of RAW264.7 cells was assessed using a CCK8 assay. The levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in the supernatant of each group were evaluated using the Griess method and an enzyme-linked immunosorbent assay (ELISA). Western blot and quantitative real-time PCR (qRT-RCR) were used to measure the levels of cyclooxygenase 2 (COX-2) and nitric oxide synthase (iNOS) gene expression. Our findings revealed that these two compounds inhibited the high levels of NO, TNF-α, IL-6, IL-1β, COX-2, and iNOS (induced by LPS). Mechanistic studies demonstrated that these two compounds reduced the activation of the nuclear transcription factor-B (NF-κB) signaling pathway by inhibiting the phosphorylation of p65. Therefore, our study indicates that compounds 1 and 2 can exert a definite anti-inflammatory effect by inhibiting the NF-κB signaling pathway.

Effect of dietary arginine supplementation on protein synthesis, meat quality and flavor in growing lambs

This study aimed to assess the effect of dietary arginine supplementation on protein synthesis, meat quality and flavor in lambs. Eighteen Dorper (♂) × Small Tailed Han sheep (♀) crossed ewe lambs of similar weight (27.29 ± 2.02 kg; aged 3 months) were assigned to two groups, the control group was fed the basal diet (Con group), and the arginine group (Arg group) was supplemented with 1% l-arginine based on the Con group for 90 d. The results suggested that dietary arginine significantly increased final body weight, loin eye muscle area, muscle fiber diameter, cross-sectional area (P < 0.050), and decreased shear force value and cooking loss (P < 0.050), as well as altered the composition and contents of volatile flavor compounds in lambs. Importantly, the total protein (TP) content, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP) activities in serum, branched-chain aminotransferase (BCAT), AST, ALT activities and neuronal nitric oxide synthase (nNOS) gene expression and content were elevated (P < 0.050), while content of urea nitrogen (BUN) in serum and 3-methylhistidine (3-MH) were decreased in arginine fed lambs (P < 0.050). In addition, arginine triggered muscle protein synthesis through protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway, while minimized protein degradation by regulating gene expression of myogenin (MyoG), myostatin (MSTN), muscle atrophy F-box (MAFbx) and forkhead box O3 family (FoxO3) (P < 0.050). Taken together, this study suggested that arginine can be used to improve protein deposition and meat quality in lamb production.

Multifactorial model for prediction of the development of polymorphic ventricular tachycardia in patients with drug-induced QT interval prolongation induced by class III antiarrhythmic drugs

Aim. To develop a multifactorial model for predicting the development of polymorphic ventricular tachycardia (VT) in patients with drug-induced long QT syndrome (LQTS) induced by class III antiarrhythmic drugs (AADs) by identifying electrocardiographic, laboratory and molecular genetic predictors.Methods. The study included 64 patients (37 (57.9%) women and 27 (42.1%) men, mean age 57.2±9.4 years) with ischemic heart disease and/or arterial hypertension. and cardiac arrhythmias, in which drug-induced prolongation of the QTc interval (Bazett) (over 450 ms in men and over 470 ms in women) was noted with the use of class III AADs (amiodarone or sotalol) in a cardiac hospital. Depending on the presence or absence of non-sustained polymorphic VT according to 24-hour ECG Holter monitoring, patients were further divided into two groups: 17 patients with episodes of non-sustained polymorphic VT and 47 patients without such episodes. All patients underwent clinical and laboratory, instrumental and molecular genetic studies, which included taking an anamnesis, recording ECG in 12 leads, biochemical blood test, determining the levels of neuronal NO-synthase (NOS1) and the adapter protein of neuronal NO-synthase (NOS1AP) in blood serum by enzyme immunoassay, determination of nitric oxide synthase gene polymorphisms by polymerase chain reaction. To assess the relationship of the studied parameters with the achievement of the end point, the method of logistic regression with a binary response and the logit function of the connection was used.Results. To assess the risk of developing non-sustained polymorphic VT in patients with drug-induced LQTS while taking class III AADs, a complex binary logistic regression model was developed, including the following indicators: patient gender (p=0.019), relative variance of the QT interval (p=0.002), duration of the Tpeak-Tend interval, (p=0.034), serum magnesium (p=0.004) and NOS1 (p=0.004) levels, as well as the AA genotype of the G84A polymorphism of the NOS1 gene (p=0.049). With the calculated value of the threshold probability p≥0.48, the developed model makes it possible to identify patients at high risk of developing polymorphic VT in patients with drug-induced LQTS with a sensitivity of 94.12%, a specificity of 89.36%, and an area under the ROC curve of 0.977 (0.95-1.0, p&lt;0.001).Conclusion. The developed complex model will allow predicting the risk of proarrhythmic effects in patients with drug-induced LQTS, which will lead to a decrease in the number of cardiovascular events in this category of patients.

Curcumin potentiates the anti-inflammatory effects of Tehranolide by modulating the STAT3/NF-κB signaling pathway in breast and ovarian cancer cell lines.

Studies have demonstrated thatnatural products, such as curcumin and artemisinin,possess anti-inflammatory effects, which can be beneficial for cancer treatment.Tehranolide, as a novel natural product, has a wide range of biological activities, including anti-cancer effects. However,many properties of Tehranolide, like its anti-inflammatory activity and its combination with curcumin, have not been investigated yet. This investigation examined the anti-inflammatory activity of Tehranolide, either alone or in combination with curcumin, via modulating the NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) and STAT3 (signal transducer and activator of transcription 3) signaling pathways in MDA-MB-231 and SKOV3, breast and ovarian cancer cell lines. ELISA-based methods were employed to measure the pro-inflammatory cytokine levels and the NF-κB activity in lipopolysaccharide (LPS)-induced cells. The real-time PCR experiment and Griess test were performed to evaluate inducible nitric oxide synthase (iNOS) gene expression and nitrite levels, respectively. The STAT3 and NF-κB signaling pathways were investigated by Western blotting analysis. Tehranolide's anti-cancer activity was also assessed in a mouse model of breast cancer using the TUNEL (terminal deoxynucleotidyl transferase nick-end labeling) assay. Tehranolide diminished levels of pro-inflammatory cytokines in cancer cells. Additionally, it suppressed NF-κB DNA binding and STAT3 phosphorylation, reducing iNOS gene expression and nitrite production. Moreover, Western blotting showed that Tehranolide enhanced the inhibitory κB (IκBα) and Bcl-2 (B-cell lymphoma 2)-associated X (BAX) expression, and downregulated the expression of Bcl-2 proteins. Furthermore, the TUNEL assay demonstrated that Tehranolide induced apoptosis in a breast cancer mouse model. Curcumin potentiated all the anti-inflammatory effects of Tehranolide. This investigation indicated for the first time that Tehranolide, either alone or in combination with curcumin, exerted its anti-inflammatory effects by suppressing NF-κB and STAT3 signaling pathways in SKOV3 and MDA-MB-231 cells.

Nitric oxide synthase 2 genetic variation rs2297514 associates with a decreased susceptibility to extremity post-traumatic osteomyelitis in a Chinese Han population.

Previous studies have indicated that nitric oxide synthase 2 (NOS2) genetic variations are involved in delayed fracture healing and fracture non-union. Whether these genetic variants associate with the development of osteomyelitis (OM) remains unclear. Here, we analyzed the potential relationships between NOS2 genetic variations and the risk of developing post-traumatic OM (PTOM) in a Chinese Han population. Altogether 704 participants, including 336 PTOM patients and 368 healthy controls, were genotyped of rs2297514 and rs2248814 of the NOS2 gene using the SNaPshot genotyping method. Outcomes showed that the frequency of allele C of rs2297514 in the patient group was significantly lower than that in the control group (48.7% vs. 54.5%, P = 0.029, OR = 0.792, 95% CI 0.642 - 0.976). In addition, significant associations were found between rs2297514 and susceptibility to PTOM by the recessive model (P = 0.007, OR = 0.633, 95% CI 0.453 - 0.884), and the homozygous model (P = 0.039, OR = 0.648, 95% CI 0.429 - 0.979). Moreover, patients with the CC genotype of rs2297514 had lower inflammatory biomarkers levels than the TT genotype, especially for the C-reactive protein (CRP) level (median: 4.1 mg/L vs. 8.9 mg/L, P = 0.027). However, no significant relationship was noted between rs2248814 and the risk of developing PTOM. In this Chinese cohort, rs2297514 is correlated with a decreased risk of PTOM development, with genotype CC as a protective factor.

Structural and functional condition of the heart in patients with arterial hypertension depending on A1166C-gene polymorphism of angiotensin II type 1 and T786C-promoter of endothelial NO-synthase gene

It's known that pathological changes in structural and functional condition of the heart, which are caused by arterial hypertension, trigger development of chronic heart failure and disablement of population. Nowadays, it proven that candidate genes, expression products of which participate in regulating vascular tone, have considerable influence upon development of arterial hypertension, however their role in pathogenesis of arterial hypertension has not been fully clarified, and results of the respective researches significantly vary among different populations. To determine the structural and functional state of the heart in patients with hypertension depending on the polymorphism of the A1116C gene of the angiotensin II receptor type I and the T786C promoter of the endothelial NO-synthase gene we examined 86 patients, aged from 45 to 76 years. 30 people without signs of hypertension were in the control group. The structural and functional state of the heart was assessed by cardiac ultrasound according to standard methods. Studies of the A1166C allelic polymorphism of the angiotensin II receptor gene type 1 and the T786C promoter of the eNOS gene were performed by polymerase chain reaction with electrophoretic detection of results. Analysis of cardiac ultrasound showed, that in the patients – carriers of C-allele of both studied genes (AС+СС and TC+CC) left ventricular ejection fraction tended to decrease. We found a bigger thickness of the posterior wall of the left ventricle in patients with CC genotype compared to carriers of AA genotype A1166C – (1.3±0.07) cm vs. (1.1±0.05) cm (p&lt;0.005). The mass of the left ventricular myocardium index in the group of patients with the genotype AC and CC was (157.5±7.3) g/m2 and (161.5±7.1)) g/m2, respectively, being by 16.7% and 19.6% more than in carriers of AA genotype of the AGTR1 gene. In the groups of patients-carriers of C-allele (TC+CC) of the eNOS gene the mass of the left ventricular myocardium index values were (155.2±11.4)) g/m2 and (158.4±7.9)) g/m2, respectively, which is by 5.4% and 7.5% more than in carriers of TT genotype. The mean size of the left atrium was significantly higher in patients who had AC and CC genotype of the AGTR1 gene, as well as TC and CC genotype of the eNOS gene compared with the control group. Carriers of C-allele (AC+CC genotype) of AGTR1 gene polymorphism had clearly bigger sizes of the left atrium, as compared to homozygotes by A-allele. The severity of diastolic dysfunction was higher in carriers of the CC genotype of the AGTR1 gene and the eNOS gene than in heterozygotes of the studied genes by 4.3% and 3.3%, respectively. The research shows that inheritance of CC genotype for A1166C polymorphism of the the angiotensin II type 1 receptor gene and of CC T786C polymorphism in the promoter of eNOS gene is associated with more noticeable changes in structural and functional heart condition among patients with arterial hypertension.

The eNOS-G894T genetic polymorphism and risk of preeclampsia: A case-control study, an updated meta-analysis, and a bioinformatic assay

ObjectivesPreeclampsia (PE) is a leading cause of maternal death worldwide and involves vascular endothelial dysfunction. The aim of this study was to investigate the association of the G894T polymorphism in the endothelial nitric oxide synthase (eNOS) gene and the risk of preeclampsia in a case-control design in an Iranian population, which was followed by a meta-analysis and an in silico approach. MethodsIn the case-control study, 300 people including 135 pregnant women with preeclampsia and 165 healthy pregnant women were included. The genotype of G894T polymorphism was determined by the PCR-RFLP method. We searched authoritative scientific databases to find eligible studies for meta-analysis. The odds ratio with a 95% confidence interval was estimated to find the strength of the association of the mentioned polymorphism with the risk of preeclampsia. In addition, the effect of G894T transversion on eNOS gene function was evaluated by some bioinformatics tools. ResultsOur case-control data showed that the G894T polymorphism is associated with an increased risk of preeclampsia. In the meta-analysis, 33 eligible studies were included, and the results showed that the G894T polymorphism is associated with an increased risk of preeclampsia in the overall analysis and some stratified analyses. In addition, the structural analysis showed that the G894T variant can affect the splicing process as well as the protein stability. ConclusionsBased on the results, the aforementioned polymorphism may be a risk factor for preeclampsia and could be considered a potential molecular biomarker for screening susceptible individuals.

Genetic origins of recurrent miscarriage: A review of the literature

Currently, recurrent miscarriage is considered a polygenic multifactorial disease, that is, a condition in which the predisposition to miscarriage is related to various genetic factors. The realization of these factors depends largely on the environment. This paper analyzes the current world literature on the effect of single-nucleotide polymorphisms of genes associated with recurrent miscarriage. The literature review summarized data from recent meta-analyses regarding the association between recurrent miscarriage and genetic polymorphisms leading to endothelial dysfunction (vascular endothelial growth factor A gene and endothelial NO synthase genes), hereditary thrombophilia (Leiden mutation and polymorphisms in the methylene tetrahydrofolate reductase gene), immunological disorders (cytokine genes, including interleukin-1 beta, interleukin-6, and tumor necrosis factor alpha), altered receptor interactions in the endometrium (progesterone receptor and ESR1 and ESR2 estrogen receptor genes), and imbalance between lipoproteins of different densities (apolipoprotein E gene). The study of genetic predisposition to miscarriage is necessary to identify high-risk cases, initiate timely and careful monitoring, and develop personalized preventive techniques.

ENOS polymorphisms on male infertility: An updated systematic review and meta-analysis.

This meta-analysis was performed to examine the association of 3 endothelial nitric oxide synthase (eNOS) gene polymorphisms with male infertility. The literature on the relation between the mutant of eNOS and male infertility before July 1, 2022, was conducted in Pubmed, Medline, and Web of Science. The search strategy is as follows: (eNOS OR ECNOS OR nitric oxide synthase 3 OR NOS3) AND (polymorphism OR mutation OR variation OR SNP OR genotype) AND (male infertility). Statistical analysis was performed with the web of MetaGenyo, Stata 12, trial sequential analysis 0.9Beta, and the web of GTEx. Overall, 13 studies (26 case-controls) were included involving 6518 cases and 5461 controls for 3 polymorphisms (rs2070744, rs1799983, rs61722009) of eNOS. We found that eNOS rs2070744 was correlated with an increased risk of male infertility (C vs. T: odds ratio [OR], 1.48; 95% confidence interval [CI], [1.19-1.85]; CC vs. TT: OR, 2.59; 95% CI, [1.40-4.80]; CT vs. TT: OR, 1.17; 95% CI, [1.00-1.38]; CC vs. CT + TT: OR, 2.50; 95% CI, [1.35-4.62]; CC + CT vs. TT: OR, 1.41; 95% CI, [1.21-1.64]). And eNOS rs1799983 was correlated with an increased risk of male infertility (allele contrast T vs. G: OR, 1.41; 95% CI, [1.01-1.96]; P = .043; recessive model TT vs. TG + GG: OR, 2.00; 95% CI, [1.03-3.90]; P = .042). In the stratified analysis of rs61722009, we found Asians might be correlated with an increased risk of male infertility (4a vs. 4b: OR, 1.50; 95% CI, [0.94-2.38]; 4a4a vs. 4b4b: OR, 2.56; 95% CI, [0.70-9.38]; 4a4b vs. 4b4b: OR, 1.36; 95% CI, [0.87-2.13]; 4a4a vs. 4a4b + 4b4b: OR, 2.57; 95% CI, [0.91-7.30]; 4a4a + 4a4b vs. 4b4b: OR, 1.44; 95% CI, [0.87-2.40]). The eNOS rs2070744 polymorphism and rs1799983 are associated with the risk of male infertility, and rs61722009 might be a risk factor for Asians.

DYSLIPIDEMIA AND HYPERGLYCEMIA IN HYPERTENSIVE PATIENTS ASSOCIATE WITH GNB3 (RS5443) AND NOS3 (RS2070744) GENES’ ALLELIC STATE

Objective: More than 700 million people worldwide live with untreated essential arterial hypertension (EAH). The objective of the study was to evaluate the risks of metabolic disorders in EAH patients depending on guanine nucleotide-binding protein-↓3 subunit (GNB3, rs5443) and endothelial nitric oxide synthase (NOS3, rs2070744) genes’ allelic state. Design and method: The cohort case-control study involved 100 EAH patients (moderate-very high cardiovascular risks, aged 45-65 years) and 48 practically healthy (control). Metabolic disorders were evaluated by glucose blood value and lipids panel: total cholesterol (TC), triglycerides (TG), Low-, and High- density lipoprotein cholesterol (LDL-C, HDL-C) levels. The atherogenic index (AI) was calculated by the equation: (TC-HDL-C)/HDL-C. GNB3 (rs5443) and NOS3 (rs2070744) genotyping perfrmed by TaqMan probes in CFX96 Real-Time PCR Detection System. Results: Hyperglycemia (&gt;6.1 mmol/l), hypertriglyceridemia (TG&gt;1.7 mmol/l) and decreased HDL-C (&lt;1.2 mmol/l) were relatively more common in EAH patients than in the control group by 36.11% (⇙2 = 17.88; p&lt;0.001), 23.61% (⇙2 = 6.43; p = 0.011) and 25.0% (⇙2 = 8.32; p = 0.004) respectively. Therefore, the fasting hyperglycemia increases the EAH risk in the examined population ninefold [OR95%CI:2.86-27.08; p&lt;0.001], hypertriglyceridemia and decreased HDL-C elevates the risk almost 3 and 3.5 times as well [OR95%CI:1.23-5.56; p = 0.009 and OR95%CI:1.46-8.71; p = 0.003], respectively. The risk of metabolic disorders (dyslipidemia and hyperglycemia) in EAH patients does not depend on NOS3 gene polymorphism (rs2070744). Contrary, in T-allele patients of the GNB3 gene (rs5443) prevailed subjects with elevated LDL-C (&gt;3.0 mmol/l) over those with CC-genotype by 13.89% (p = 0.05). Other parameters of lipid metabolism and hyperglycemia did not differ significantly between GNB3 (rs5443) genes allelic state. However, the mutational T-allele of the GNB3 gene (825C&gt;T) increases the risk of hyperlipidemia 8.5 times [OR 95%CI:0.99-72.70; p = 0.05] due to atherogenic LDL-C, with the protective role of CC-genotype [OR = 0.12; OR 95%CI:0.01-1.0; p = 0.048]. Conclusions: Fasting hyperglycemia, hypertriglyceridemia and lowered HDL-C, enhance the arterial hypertension risk 3-9 times (p&lt;0.01). The polymorphic site of GNB3 (rs5443) gene, but not NOS3 (rs2070744) gene associate with hyperlipidemia in hypertensive patients.

ENDOTHELIAL NITRIC OXIDE SYNTHASE (NOS3) AND GUANINE NUCLEOTIDE-BINDING PROTEIN (GNB3) GENES’ POLYMORPHISMS PORTEND OBESITY IN HYPERTENSIVE PATIENTS: THE GENDER ASPECT

Objective: Obesity is a major risk factor of essential arterial hypertension (EAH) because of neuro-hormonal, sympathoadrenal and renin-angiotensin-aldosterone system activation, as well as endothelial dysfunction and systemic low grade inflammation. However, genetic mechanisms of obesity development in EAH patients remained unclear. Therefore, we studied the guanine nucleotide-binding protein-↓3 (GNB3, rs5443) and endothelial nitric oxide synthase (NOS3, rs2070744) genes’ polymorphisms as possible genetic harbingers of obesity in EAH patients particularly focusing on gender. Design and method: One-hundred EAH patients with hypertensive-mediated organ damage (moderate-very high cardiovascular risks, aged 45-65 years) and 48 practically healthy (control) participated in the cohort case-control study. Obesity was determined by body mass index (BMI) &gt;30 kg/m2. GNB3 (rs5443) and NOS3 (rs2070744) genotyping performed by TaqMan probes (CFX96 Real-Time PCR). Results: The NOS3 gene mutation in the homozygous state occurs in 16.67% of EAH patients and for the GNB3 gene in 8.33% of cases, which does not differ from the control group. The relative frequency of obese prevailed among EAH patients with the mutated C-allele carriers of the NOS3 gene by 31.94% (⇙2 = 13.58; p&lt;0.001) and T-allele of the GNB3 gene (30.56%) in the absence of such among healthy control. The risk of obesity increases in EAH patients in C-allele carriers of the NOS3 gene almost 6 times [OR 95%CI:2.11-14.82; p&lt;0.001] and in T-allele patients of the GNB3 gene – more than 10 times [OR 95%CI:2.25-45.44; p&lt;0.001], 1.3 times more often in women than men (p&lt;0,05), regardless the genes’ allelic state. The TT-genotype of the NOS3 gene and the CC-genotype of the GNB3 gene play a protective role against obesity. Conclusions: The C-allele of the NOS3 gene (rs2070744) and the T-allele of the GNB3 gene (rs5443) increase the obesity risk in hypertensive patients 6-10 fold (p&lt;0.001) as well, 1.3 times more often in women, than men.

Effect of treatment on genetic predictors and quality of life in rheumatoid arthritis patients with type 2 diabetes, hypertension, and obesity

Background. Connective tissue disease, in particular rheumatoid arthritis (RA), is characterized by a progressive joint damage and polymorphism of extra-articular lesions that affect the patient’s quality of life. Although the widespread implementation of basic therapy through the use of disease-modifying antirheumatic drugs has a positive effect on social consequences, the experience gained shows the impossibility of achieving a stable remission of the disease or reducing the activity of autoimmune inflammation against the background of using these medicines in all patients for a number of reasons. One of these reasons is the presence of comorbid pathology, which requires a personalized approach to each person. The objective was to study the quality of life of patients with RA in combination with type 2 diabetes mellitus (T2DM), hypertension and obesity depending on the T-786C polymorphism of the endothelial nitric oxide synthase gene promoter. Materials and methods. A laboratory and instrumental examination involved 110 patients who were treated at the clinical base of the Department of Internal Medicine of Bukovinian State Medical University, Chernivtsi Regional Endocrinology Center, Chernivtsi Regional Clinical Hospital, and Reference Center for Molecular Genetic Research of the Ministry of Health of Ukraine. Results. When assessing the quality of life in the studied groups, a decrease in indicators on all scales was detected. Thus, in RA combined with hypertension and obesity, PF was 1.73 times lower, RP— 1.97 times, BP— 1.44 times, SF— 1.46 times, MH— 1.26 times , RE— 1.66 times, GH— 1.35 times (p&lt;0.05) lower than in patients with isolated RA. Taking into account the T-786C polymorphism of the eNOS gene, it can be asserted that all indicators of quality of life were reduced in the carriers of the СС genotype, in contrast to the carriers of the T-allele: PF— by 2.06 (pTT&lt;0.05) and 2.46 times (pTС&lt;0.05); RP— by 2.0 (pTT&lt;0.05) and 2.87 times (pTС&lt;0.05); BP— by 1.86 (pTT&lt;0.05) and 2.52 times (pTС&lt;0.05); SF— by 1.55 and 2.07 times (рTС&lt;0.05); MH— by 1.42 (pTT&lt;0.05) and 1.53 times (pTС&lt;0.05); RE— by 1.30 (pTT&lt;0.05) and 1.54 times (pTС&lt;0.05); VT— by 3.34 (pTT&lt;0.05) and 3.72 times (pTС&lt;0.05); GH— by 2.32 (pTT&lt;0.05) and 2.38 times (pTС&lt;0.05), respectively. Conclusions. When comparing the quality of life in the studied groups, it can be stated that the lowest indicators on all scales were in patients with RA combined with hypertension, obesity and T2DM and in carriers of the СС genotype of the analyzed gene, which reflected the worst physical condition and psychosocial status.

Association of endothelial nitric oxide synthase (Glu298Asp) gene polymorphism with radial artery spasm during cardiac catheterization in Egyptians

BackgroundNitric oxide (NO) exerts diverse effects on the cardiovascular system. Impairment of NO production plays a key role in cerebral and coronary artery spasm. We aimed to explore the predicting factors of radial artery spasm (RAS) and the association of eNOS gene polymorphism (Glu298Asp) with RAS during cardiac catheterization.Methods and results200 patients underwent elective coronary angiography through a trans-radial approach. The subjects were genotyped to the Glu298Asp polymorphism (rs1799983) on the eNOS gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our results showed that the subjects with the TT genotype and T allele were significantly more likely to develop radial artery spasms (OR = 12.5, 4.6, P < 0.001 respectively). TT genotype of eNOS Glu298Asp polymorphism, number of punctures, size of the radial sheath, radial tortuosity, and right radial access are independent predictors of radial spasm.ConclusionThe eNOS (Glu298Asp) gene polymorphism is associated with RAS during cardiac catheterization in Egyptians. TT genotype of eNOS Glu298Asp polymorphism, number of punctures, size of the radial sheath, right radial access, and tortuosity are independent predictors of RAS during cardiac catheterization.

Synergistic Immunostimulatory Activities of Probiotic Strains, Leuconostoc lactis and Weissella cibaria, and the Prebiotic Oligosaccharides They Produce.

Synbiotics contain health-beneficial bacteria, i.e., probiotics and prebiotics selectively utilized by the probiotics. Herein, three probiotic strains, Leuconostoc lactis CCK940, L. lactis SBC001, and Weissella cibaria YRK005, and the oligosaccharides produced by these strains (CCK, SBC, and YRK, respectively) were used to prepare nine synbiotic combinations. Macrophages (RAW 264.7) were treated with these synbiotic combinations and the corresponding lactic acid bacteria and oligosaccharides alone to evaluate the treatments' immunostimulatory activities. The level of nitric oxide (NO) production was significantly higher in the macrophages treated with the synbiotics than in those treated with the corresponding probiotic strains and the oligosaccharide alone. The immunostimulatory activities of the synbiotics increased regardless of the probiotic strain and the type of oligosaccharide used. The expressions of tissue necrosis factor-α, interleukin-1β, cyclooxygenase-2, inducible NO synthase genes, and extracellular-signal-regulated and c-Jun N-terminal kinases were significantly higher in the macrophages treated with the three synbiotics than in those treated with the corresponding strains or with the oligosaccharides alone. These results indicate that the synergistic immunostimulatory activities of probiotics and the prebiotics they produced in the studied synbiotic preparations resulted from the activation of the mitogen-activated protein-kinase-signaling pathway. This study suggests the combined use of these probiotics and prebiotics in the development of synbiotic preparations as health supplements.