Low Levels Of Nitric Oxide Research Articles

Low endogenous NO levels in roots and antioxidant systems are determinants for the resistance of Arabidopsis seedlings grown in Cd

Cadmium (Cd), which is a toxic non-essential heavy metal capable of entering plants and thus the food chain, constitutes a major environmental and health concern worldwide. An understanding of the tools used by plants to overcome Cd stress could lead to the production of food crops with lower Cd uptake capacity and of plants with greater Cd uptake potential for phytoremediation purposes in order to restore soil efficiency in self-sustaining ecosystems. The signalling molecule nitric oxide (NO), whose function remains unclear, has recently been involved in responses to Cd stress. Using different mutants, such as nia1nia2, nox1, argh1-1 and Atnoa1, which were altered in NO metabolism, we analysed various parameters related to reactive oxygen and nitrogen species (ROS/RNS) metabolism and seedling fitness following germination and growth under Cd treatment conditions for seven days. Seedling roots were the most affected, with an increase in ROS and RNS observed in wild type (WT) seedling roots, leading to increased oxidative damage and fitness loss. Mutants that showed lower NO levels in seedling roots under Cd stress were more resistant than WT seedlings due to the maintenance of antioxidant systems which protect against oxidative damage.

Nitric Oxide Gene Polymorphism is a Risk Factor for Diabetic Nephropathy and Atherosclerosis in Type 1 Diabetic Patients.

AIM:To assess the risk factor for diabetic atherosclerosis nephropathy and diabetic nephropathy in type 1 diabetic patients.PATIENTS AND METHODS:Thirty healthy volunteers age and sex-matched and Sixty-five type 1 diabetic patient were in rolled in the study. The mean age of patients was 17.99 ± 2.59 years, mean age of onset of diabetes was 7.00 ± 3.28 years, mean duration of diabetes was 10.91 ± 3.54 years. Glycosylated sex-matched (HbA1c) was assessed in blood samples, serum lipid profile was determined, and serum level of oxidised low-density lipoprotein (OxLDL), and nitric oxide was evaluated by enzyme-linked immunosorbent assay (ELISA) technique. Nitric oxide 894G > T genotype was analysed by (PCR-RFLP) method and confirmed by Sequencing. Assessment of the albumin / creatinine ratio was done in urine samples. Renal Doppler and Carotid intima-media thickness (cIMT) via ultrasound was also performed.RESULTS:OxLDL, lipid profile, albumin/creatinine ratio, cIMT and resistivity index were significantly higher in diabetic patients while nitric oxide was significantly lower. Nitric oxide genotype shows no significant difference between diabetic’s patients and controls. Diabetic patients with homozygous NO had a significantly lower serum level of Nitric oxide, a significantly higher OxLDL, albumin / creatinine ratio and lipid profile.CONCLUSION:diabetic patients are liable for the occurrence of early diabetic nephropathy and atherosclerosis as a result of the presence of low level of nitric oxide. Nitric oxide gene polymorphism 894G > T in diabetic patients is a risk factor for diabetic nephropathy and atherosclerosis.

Effect of ulinastatin on myocardial ischemia-reperfusion injury through JNK and P38 MAPK signaling pathways.

The aim of this study was to investigate the effect of ulinastatin (UTI) on myocardial ischemia-reperfusion injury (MIRI) through the c-Jun N-terminal kinase (JNK) signaling pathway and p38 mitogen-activated protein kinase (MAPK) signaling pathway. Healthy adult male Sprague-Dawley (SD) rats were randomly divided into 5 groups, including the sham group (n=10), MIRI group (model group, n=10), UTI group (n=10), UTI + JNK inhibitor SP600125 (UTI+SP600125 group, n=10) and UTI + p38 MAPK inhibitor SB203580 (UTI+SB203580 group, n=10). Hemodynamics, myocardial infarction and the messenger ribonucleic acid (mRNA) expressions of p38 MAPK, JNK, superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and high-sensitivity C-reactive protein (hs-CRP) were compared among groups. The protein levels of p38 MAPK and JNK in serum were detected via Western blotting. Furthermore, the correlations of serum p38 MAPK and JNK with TNF-α were analyzed. In the UTI group, the left ventricular systolic pressure (LVSP), the left ventricular end-diastolic pressure (LVEDP) and maximal rate of increase of the left ventricular pressure (+dp/dtmax) were significantly higher than those of the model group. However, the maximal rate of the decrease of the left ventricular pressure (-dp/dtmax), infarction area and ischemia area were significantly smaller than those of the model group. LVSP, LVEDP and +dp/dtmax in UTI+SP600125 group and UTI+SB203580 group were markedly higher than those of the UTI group. Meanwhile, the mRNA expressions of p38 MAPK and JNK in the UTI group were significantly lower than those of the model group. However, the mRNA expression levels of p38 MAPK and JNK in UTI+SP600125 group and UTI+SB203580 group were remarkably higher than the UTI group. Besides, the UTI group showed a markedly higher level of SOD and significantly lower levels of MDA, NO, TNF-α, IL-6 and hs-CRP than the model group. Furthermore, UTI+SP600125 group and UTI+SB203580 group showed significantly higher levels of MDA, NO, TNF-α, IL-6 and hs-CRP than the UTI group. The protein levels p38 MAPK and JNK were remarkably lower in the UTI group than those of the model group. However, they were remarkably higher in UTI+SP600125 group and UTI+SB203580 group than the UTI group. In addition, both p38 MAPK and JNK proteins were positively correlated with TNF-α (r=0.983 and 0.892, p=0.043 and p=0.033). UTI may inhibit MIRI caused by p38 MAPK signaling pathway and JNK signaling pathway by down-regulating TNF-α expression, thereby protecting and improving MIR.

Cytoplasmic "ciliary inclusions" in isolation are not sufficient for the diagnosis of primary ciliary dyskinesia.

The diagnosis of primary ciliary dyskinesia (PCD) is difficult and requires a combination of clinical features, nasal nitric oxide testing, cilia ultrastructural analysis by electron microscopy (EM), and genetics. A recently described cytoplasmic ultrastructural change termed "ciliary inclusions" was reported to be diagnostic of PCD; however, no supporting evidence of PCD was provided. In this study, we sought to confirm, or refute, the diagnosis of PCD in subjects with "ciliary inclusions" on EM. Six subjects from five families with previous lab reports of "ciliary inclusions" on EMs of ciliated cells were identified and evaluated at a Genetic Disorders of Mucociliary Clearance Consortium site. We performed a detailed clinical history, nasal nitric oxide measurement, genetic testing including whole-exome sequencing (WES), and when possible, repeat ciliary EM study. Only one of six subjects had multiple and persistent clinical features congruent with PCD. No subject had situs inversus. Only one of six subjects had a very low nasal nitric oxide level. No "ciliary inclusions" were found in three subjects who had a repeat ciliary EM, and ciliary axonemal ultrastructures were normal. Genetic testing, including WES, was negative for PCD-causing genes, and for pathogenic variants in gene pathways that might cause "ciliary inclusions," such as ciliary biogenesis. "Ciliary Inclusions", in isolation, are not sufficient to diagnosis PCD. If seen, additional studies should be done to pursue an accurate diagnosis.

The adipokine vaspin reduces apoptosis in human hepatocellular carcinoma (Hep-3B) cells, associated with lower levels of NO and superoxide anion

BackgroundAmong adipose-derived factors, adipocytokines play roles as hormones and signaling mediators for apoptotic pathway. Among of them, vaspin, regulates the metabolism of adipose tissue itself as an endocrine organ, and stimulates adipocytes to maturation, differentiation, etc. Damaged adipocytes, present in obesity and hepatocellular carcinoma (HCC) respond with over-production of inflammatory cytokines. Such pro-inflammatory stimulation remains under adipokine control. Pro-inflammatory pathways are connected to oxidative stress and apoptosis, reported as co-existing with an elevated level of some adipokines in cancer cell lines. However, some hormones, such as vaspin, reduce apoptosis, have anti-inflammatory and anti-oxidative roles in cancer cell lines.MethodsHep-3B cells were cytometrically evaluated under vaspin treatment for reactive oxygen species (ROS) and apoptosiss induction. The statistical significant changes to the untreated controls was calculated by T-tests (indicated at value p < 0.05).ResultsHere we studied the production of reactive oxygen and nitrogen species in cells of HCC line Hep-3B after vaspin treatment. A decreased level of nitric oxide and superoxide anion 24 h after vaspin addition at 5 ng/ml was correlated with restricted, to the physiological level, apoptosis. A protective role of vaspin was displayed as enhanced cell viability and proliferation, which could be a poor prognostic in liver cancer.ConclusionsApoptosis was suppressed after vaspin treatment, together with low levels of nitric oxide and superoxide anions.

Ellisras Longitudinal Study 2017: elevated serum levels of carboxymethyl-lysine, an advanced glycation end-product, are associated with higher odds of developing endothelial dysfunction in black South African patients with type 2 diabetes mellitus (ELS 29).

This case-control study investigated the association between major types of serum advanced glycation end-products (AGEs) and selected serum/plasma markers of endothelial dysfunction in black patients with type 2 diabetes mellitus at Dr George Mukhari Academic Hospital. Serum AGEs were measured using either enzyme-linked immunosorbent assay (ELISA) or spectrofluoremetry. Serum markers of endothelial dysfunction were measured using either ELISA or calometry. The correlation and associations between major types of serum AGEs and markers of endothelial dysfunction were investigated using the Spearman correlation coefficient and bivariate logistic regression analysis, respectively. Although both serum total immunogenic AGEs and serum carboxymethyl-lysine (CML) were moderately and negatively associated with endothelial dysfunction, only serum CML was significantly associated with a higher odds for the development of endothelial dysfunction (low nitric oxide levels) in our diabetic subjects. It can therefore be concluded from this study that high serum levels of CML may predispose to endothelial dysfunction in black South Africans with type 2 diabetes.

Abstract P3047: Soluble Guanylyl Cyclase Stimulators And Activators Inhibit Hypoxia-induced Placental Sflt-1 Levels

Preeclampsia (PE) is a pregnancy-specific disease characterized by placental ischemia, maternal endothelial dysfunction, and hypertension. The ischemic placenta releases anti-angiogenic factors, such as sFlt-1, which antagonizes the angiogenic factors, Placental Growth Factor (PlGF) and Vascular Endothelial Growth Factor (VEGF), causing endothelial dysfunction and impaired nitric oxide (NO) signaling. During normal pregnancy, NO would bind soluble guanylyl cyclase (sGC) increasing cGMP levels leading to vasodilation. However, in PE this signaling is reduced. sGC stimulators and activators are a novel class of drug that directly stimulate sGC despite low levels of NO. Our preliminary data suggest that sGC modulators reduce hypertension in an animal model of PE, but the mechanisms are not known. Therefore, this study tested the hypothesis that sGC stimulators (Riociguat) and activators (Cinaciguat) attenuate sFlt-1 production and increase PlGF levels in the placenta. Placentas were obtained from normal pregnant rats on gestational day 19. Isolated placental villi were plated (1 per well) and coated with Matrix Matrigel Basement Membrane. The villi were treated with various doses of sGC modulators (0.2μM, 1μM, 10μM, 30μM) and then exposed to a hypoxic environment (1% oxygen) for 48 hours. Levels of sFlt-1, PlGF, and VEGF were determined using an Enzyme-Linked Immunosorbent Assay kit.sFlt-1 concentration was significantly reduced in both groups treated with the stimulator or activator compared to the untreated group. The greatest attenuation was observed with the 30μM sGC activator treatment (Untreated, 5117±1313 pg/mL; 30μM sGC stimulator, 3975±621pg/mL, 30μM sGC activator, 3811±712 pg/mL, P&lt;0.05). PlGF was not significantly different between groups, however, VEGF concentrations were significantly increased at higher doses (Untreated, 3099±100 pg/mL; 30μM sGC stimulator, 3871±449 pg/mL 30μM sGC activator, 3952±252 pg/mL; p&lt;0.05). These data suggest that sGC modulators significantly attenuate sFlt-1 and thus, improve the sFlt-1/PlGF and sFlt-1/VEGF ratios. Therefore, a potential mechanism whereby sGC modulators reduce blood pressure in PE is by inhibiting hypoxia-induced placental sFlt-1 levels.

Cooperative Roles of Nitric Oxide-Metabolizing Enzymes To Counteract Nitrosative Stress in Enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli (EHEC) has at least three enzymes, NorV, Hmp, and Hcp, that act independently to lower the toxicity of nitric oxide (NO), a potent antimicrobial molecule. This study aimed to reveal the cooperative roles of these defensive enzymes in EHEC against nitrosative stress. Under anaerobic conditions, combined deletion of all three enzymes significantly increased the NO sensitivity of EHEC determined by the growth at late stationary phase; however, the expression of norV restored the NO resistance of EHEC. On the other hand, the growth of Δhmp mutant EHEC was inhibited after early stationary phase, indicating that NorV and Hmp play a cooperative role in anaerobic growth. Under microaerobic conditions, the growth of Δhmp mutant EHEC was inhibited by NO, indicating that Hmp is the enzyme that protects cells from NO stress under microaerobic conditions. When EHEC cells were exposed to a lower concentration of NO, the NO level in bacterial cells of Δhcp mutant EHEC was higher than those of the other EHEC mutants, suggesting that Hcp is effective at regulating NO levels only at a low concentration. These findings of a low level of NO in bacterial cells with hcp indicate that the NO consumption activity of Hcp was suppressed by Hmp at a low range of NO concentrations. Taken together, these results show that the cooperative effects of NO-metabolizing enzymes are regulated by the range of NO concentrations to which the EHEC cells are exposed.

Abstract 1186: Regulation of inducible nitric oxide synthase at the single cell level modulates the inflammatory microenvironment

Abstract The anti-cancer versus pro-tumor behavior of cancer tissues is dependent largely on tumor cell-macrophage interactions in the tumor/inflammatory microenvironment and is regulated by Nitric Oxide (NO). High inducible nitric oxide synthase (Nos2) is associated with poor prognosis in breast cancer. Previously, we found that murine macrophages can be activated by different inflammatory cytokines/LPS to produce distinct NO fluxes (entirely Nos2-derived), suggesting flux-specific biological ramifications of NO. However, the effects of these NO fluxes on M1 phenotype have not been delineated. LC/MS analysis of M1 stimulated, wild type (wt) and Nos2-/- macrophages showed that Nos2 was the only source of citrulline. Metabolic analyses and microscopy showed that flattened cell phenotype characteristic of M1 macrophages and mitochondrial respiration are Nos2 dependent and regulated in a NO flux-dependent manner, while proinflammatory cytokine profile and aerobic glycolysis are Nos2 independent. We show for the first time that induction of Nos2 expression occurs only in specific stimulated cells that also harbor depolarized mitochondria. NO production has been linked to decreased oxygen consumption in hypoxic environments. We utilized a novel, in vitro chamber system that forms cell-generated hypoxic and metabolic gradients in two-dimensions by restricting the diffusive exchange of oxygen and metabolites to a monolayer of cells in a small volume- analogous to diffusion between a capillary and nearby tissue. We investigated interactions between Nos2 in M1-stimulated macrophages and hypoxia and demonstrated that treatment with IFNγ+LPS increases Nos2 expression and alters the magnitude and spatial extent of hypoxic gradients. A modified scratch assay revealed that low doses (1-50μM) of NO increased and high doses (1000μM) inhibited the migratory capacity of 4T1 tumor cells. However, in vivo, Nos2-/- mice did not show difference in primary tumor or metastatic burden compared to wt mice but bone marrow derived macrophages (BMDMs) from wt tumor bearing mice produced significantly lower levels of NO compared to BMDMs from tumor bearing- Nos2-/- mice. In summary, we find that the right flux NO is required to tune the inflammatory microenvironment. Nos2 and citrulline are robust intracellular readouts of extracellular NO flux. Nos2 dependent and independent events cooperate to regulate inflammatory macrophages. Autocrine, single cell effects on metabolism build up to cause paracrine effects including alleviation of hypoxia. 4T1 primary tumor and lung metastasis were not Nos2-regulated but host Nos2 hampered the ability of BMDMs to respond to proinflammatory stimuli hinting at possible systemic effects of Nos2. Hence, to make a reliable prognostic prediction, it is important to know the exact NO flux, which cells within the tumor express Nos2 and what other cells associate with Nos2hi cells. Citation Format: Veena Somasundaram, Caroline Gilmore, Erika M. Palmieri, Debashree Basudhar, Will Heinz, Robert Y. Cheng, Lisa A. Ridnour, Stephen J. Lockett, Daniel W. McVicar, David A. Wink. Regulation of inducible nitric oxide synthase at the single cell level modulates the inflammatory microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1186.

Stimulation of Macrophage Cells Against Cutaneous Leishmaniasis Using Silver Nanoparticles

Cutaneous leishmaniasis is a disease caused by Leishmania tropica parasite. Current treatments for this parasite are undesirable because of their toxicity, resistance, and high cost. Macrophages are key players against pathogens. Nitric oxide (NO), a molecule produce by immune cells, controls intracellular killing of pathogens during infection. Silver nanoparticles (Ag NPs) demonstrated broad-spectrum activity against various types of infectious diseases. It has the ability to stimulate oxygen species production. This study aims to analyze the macrophages activation through NO production and estimate the cytotoxicity based on the lactate dehydrogenase (LDH) release upon exposure to L. tropica and Ag NPs. Serially concentrations of Ag NPs were used under two conditions during and following macrophages exposure to L. tropica. MTT assay was used to determine the cytotoxicity of Ag NPs on L. tropica amastigotes during infection of macrophages in vitro. The results showed that by increasing the Ag NPs concentrations, the viability percentage of L. tropica amastigotes decreased and reached to 21.7 ± 0.64 % during infection compared with the control. The 50% inhibitory concentration of Ag NPs on amastigotes was 2.048µg/ml during infection. Moreover, post-phagocytosis study involved the assessment of NO and LDH release by macrophages upon exposure to L. tropica. It have shown that untreated macrophages released low levels of NO while in the presence of Ag NPs, macrophages were activated to produce higher levels of NO under all experimental conditions. On the other hand, macrophages were capable of controlling cytotoxicity and decreasing LDH levels during phagocytosis of L. tropica amastiogotes. Taking together, these findings suggest that Ag NPs can enhance macrophages NO production which provides a method for the identification of Ag NPs ligands with microbicidal and anti-cytotoxic properties against L. tropica pathogens.

Stimulation of Macrophage Cells Against Cutaneous Leishmaniasis Using Silver Nanoparticles

Cutaneous leishmaniasis is a disease caused by Leishmania tropica parasite. Current treatments for this parasite are undesirable because of their toxicity, resistance, and high cost. Macrophages are key players against pathogens. Nitric oxide (NO), a molecule produce by immune cells, controls intracellular killing of pathogens during infection. Silver nanoparticles (Ag NPs) demonstrated broad-spectrum activity against various types of infectious diseases. It has the ability to stimulate oxygen species production. This study aims to analyze the macrophages activation through NO production and estimate the cytotoxicity based on the lactate dehydrogenase (LDH) release upon exposure to L. tropica and Ag NPs. Serially concentrations of Ag NPs were used under two conditions during and following macrophages exposure to L. tropica. MTT assay was used to determine the cytotoxicity of Ag NPs on L. tropica amastigotes during infection of macrophages in vitro. The results showed that by increasing the Ag NPs concentrations, the viability percentage of L. tropica amastigotes decreased and reached to 21.7 ± 0.64 % during infection compared with the control. The 50% inhibitory concentration of Ag NPs on amastigotes was 2.048µg/ml during infection. Moreover, post-phagocytosis study involved the assessment of NO and LDH release by macrophages upon exposure to L. tropica. It have shown that untreated macrophages released low levels of NO while in the presence of Ag NPs, macrophages were activated to produce higher levels of NO under all experimental conditions. On the other hand, macrophages were capable of controlling cytotoxicity and decreasing LDH levels during phagocytosis of L. tropica amastiogotes. Taking together, these findings suggest that Ag NPs can enhance macrophages NO production which provides a method for the identification of Ag NPs ligands with microbicidal and anti-cytotoxic properties against L. tropica pathogens.

Associations of the NOS3 rs1799983 polymorphism with circulating nitric oxide and lipid levels: a systematic review and meta-analysis.

Circulating nitric oxide (NO) and lipid levels are closely associated with coronary artery disease (CAD). It is unclear whether the rs1799983 polymorphism in endothelial nitric oxide synthase (NOS3) gene is associated with plasma levels of NO and lipids. This systematic review and meta-analysis (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) aimed to clarify the relationships between the rs1799983 polymorphism and plasma levels of NO and lipids. Sixteen studies (2702 subjects) and 59 studies (14 148 subjects) were identified for the association analyses for NO and lipids, respectively. Mean difference (MD) and 95% CI were used to estimate the effects of the rs1799983 polymorphism on plasma NO and lipid levels. The primary outcome variable was NO, and the secondary outcomes included triglycerides, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C). Carriers of the T allele had lower levels of NO (MD -0.27 μmol/L, 95% CI -0.42 to -0.12 μmol/L, p<0.001) and HDL-C (MD -0.07 mmol/L, 95% CI -0.14 to -0.00 mmol/L, p=0.04), and higher levels of TC (MD 0.13 mmol/L, 95% CI 0.06 to 0.20 mmol/L, p<0.001) and LDL-C (MD 0.14 mmol/L, 95% CI 0.05 to 0.22 mmol/L, p=0.002) than the non-carriers. Triglyceride levels were comparable between the genotypes. The association between the NOS3 rs1799983 polymorphism and CAD may be partly mediated by abnormal NO and lipid levels caused by the T allele.

Long-Term Aspirin Administration Has No Effect on Erectile Function: Evidence from Adult Rats and Ageing Rat Model

As the broad spectrum pharmacological action, aspirin has been one of the most widely used medicines since its initial synthesis; however, the association between aspirin and erectile function is still controversial. We aim to explore whether long-term aspirin administration deteriorates or preserves erectile function from adult rats and ageing rat model. Twenty adult rats (10 weeks of age) and twenty ageing rats (80 weeks of age) were randomly divided into four groups as follows: Adult-Control (normal saline [NS]), Adult-Aspirin (aspirin, 10 mg/kg/d), Ageing-Control (NS), and Ageing-Aspirin (aspirin, 10 mg/kg/d) groups (n = 10 per group). For all rats, erectile function was assessed by maximum intracavernous pressure (ICP), total area under ICP curve (AUC), ICP/mean arterial pressure (MAP) ratio, and MAP. The total treatment duration was one month. Protein expression levels of cyclooxygenase-1 (COX-1), COX-2, endothelial nitric oxide synthase (eNOS), and nNOS of the corpus cavernosum were detected by Western blot. ELISA kits were used to determine 6-keto PGF1a, PGE2, TXB2, cyclic adenosine monophosphate (cAMP), and cyclic guanosine monophosphate (cGMP) levels. Total nitric oxide (NO) concentration was measured using a fluorometric assay kit. As a result, Ageing-Control rats revealed significantly decreased ICP, AUC, and ICP/MAP ratios compared to Adult-Control rats, and these effects were accompanied by reduced eNOS protein expression and lower total NO and cGMP levels; however, no difference was found in nNOS protein expression. For adult rat groups, aspirin significantly inhibited the production of 6-keto PGF1a, PGE2, and TXB2; however, it neither changed the ICP, AUC, or ICP/ MAP ratios nor altered the protein expression of eNOS, nNOS, COX-1, and COX-2. Meanwhile, aspirin did not influence the concentrations of total NO, cAMP, or cGMP. The same tendency was also found in the ageing rat model, which confirmed that aspirin did not alter erectile function. Our data suggested that long-term aspirin administration did not strengthen or weaken erectile function in adult rats or ageing rat model. Thus, it had no impact on erectile function.

A Very Uncommon Case of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) describes a rare, progressive disease of the pulmonary vasculature, involving a group of clinical conditions that result in precapillary pulmonary hypertension (PH). PAH is characterized by proliferative vasculopathy, and subsequent right heart failure. For the past several decades research has focused on identification of underlying molecular causes of this disease. Recently, there have been a number of reported cases of patients with scurvy developing pulmonary arterial hypertension. Thus, it has been hypothesized that vitamin C deficiency results in non-hypoxic activation of hypoxia inducible transcription factors (HIF) and low nitric oxide (NO) level in the pulmonary vasculature, leading to subsequent pulmonary vasculopathy and an exaggerated pulmonary vasoconstrictive response. Immediate supplementation of vitamin C is considered the definitive treatment, preventing an otherwise fatal outcome. In this paper, we describe a patient with a fatal case of scurvy related PAH, who was admitted with ecchymosis, oral ulceration, and dyspnea.

Evaluation of elabela, apelin and nitric oxide findings in maternal blood of normal pregnant women, pregnant women with pre-eclampsia, severe pre-eclampsia and umbilical arteries and venules of newborns

Pre-eclampsia is multisystem metabolic diseases, commonly accompanied by hypertension and proteinuria, which are among the important causes of maternal and perinatal mortality and morbidity worldwide. In a pre-eclampsia animal model study in the last year, Elabela (ELA) infusion was reported to correct hypertension and proteinuria and to normalise the birth weights of the offspring. Therefore, our main goal in this human study is to compare ELA, apelin (APLN) and nitric oxide (NO) levels in the maternal blood of pregnant women with pre-eclampsia and severe pre-eclampsia and in their newborns’ venous-arterial cord blood with maternal blood of healthy pregnant women and their newborns’ venous-arterial cord blood. Thirty controls, 28 pre-eclampsia and 24 severe pre-eclampsia cases and their newborns participated in this study. Maternal blood and newborn venous-arterial cord blood samples were collected from these patients. ELA, APLN and NO levels in these samples were measured by ELISA method. When the maternal blood ELA, APLN and NO amounts were compared with control groups, there was a significant decrease in both pre-eclamptic and severe pre-eclamptic women and this was more prominent in the women with severe pre-eclampsia. When ELA, APLN and NO levels in the newborn venous-arterial cord blood of control group was compared with that of severe pre-eclamptic and pre-eclamptic women; it was parallel with maternal findings. ELA, APLN and NO levels appear to play a role in the pathophysiology of pre-eclampsia. It is predicted that if these molecules, which are reduced due to pre-eclampsia and severe pre-eclampsia, are brought to physiological limits in the future; pre-eclampsia related maternal and perinatal mortality and morbidity can be reduced.Impact StatementWhat is already known on this subject? There are two studies (one human and one animal) in the literature evaluating only maternal elabela (ELA) levels in pre-eclamptic pregnancies. The animal study demonstrated decreased blood ELA levels in pre-eclamptic animals and the human study found increased blood ELA levels in pre-eclamptic patients. There are no studies evaluating maternal ELA levels in severe pre-eclampsia patients and also there are no studies evaluating maternal ELA levels in pre-eclampsia and severe pre-eclampsia patients. There are no studies evaluating newborns’ venous-arterial blood APLN and NO levels. Apelin (APLN) and nitric oxide (NO) results were controversial in pre-eclampsia and severe pre-eclampsia patients.What the results of this study add? The present study, for the first time, demonstrates that decreased blood ELA, APLN and NO levels in maternal blood of pregnant women with pre-eclampsia and severe pre-eclampsia and in their newborns’ venous-arterial blood. Furthermore, we have also demonstrated for the first time that decreased ELA, APLN and NO are also related with low birth weights.What the implications are of these findings for clinical practice and/or further research? The low levels of ELA, APLN and NO in maternal blood and newborns’ venous-arterial blood may be the result or the cause of pathologic changes in pre-eclampsia and severe pre-eclampsia. Also, ELA, APLN and NO may be new indicator parameters of systemic endothelial dysfunction together. More studies are needed to evaluate the relationship between of ELA, APLN and NO and pre-eclampsia and severe pre-eclampsia and in newborns’ venous-arterial blood.

Metformin prevention of doxorubicin resistance in MCF-7 and MDA-MB-231 involves oxidative stress generation and modulation of cell adaptation genes

Metformin was shown to sensitize multidrug resistant breast cancer cells; however, the mechanisms involved in this capacity need to be clarified. We investigated oxidative stress and inflammatory-related pathways during the induction of doxorubicin resistance in MCF-7 and MDA-MB-231 human breast cancer cells (DOX-res group), and evaluated metformin-induced cellular responses that resulted in the prevention of doxorubicin resistance (Met-DOX group). Microarray analysis demonstrated that DOX-res changed the expression of genes involved in oxidative stress (OS) and the TGF- β1 pathway. The DOX-res group presented increased thiols and reduced lipoperoxidation, increased levels of nitric oxide, nuclear NF-kB and Nrf2, and reduced nuclear p53 labelling. Analysis of the TGF-β1 signaling pathway by RT-PCR array showed that DOX-res developed adaptive responses, such as resistance against apoptosis and OS. Metformin treatment modified gene expression related to OS and the IFN-α signaling pathway. The Met-DOX group was more sensitive to DOX-induced OS, presented lower levels of nitric oxide, nuclear NF-kB and Nrf2, and increased nuclear p53. Analysis of the IFN-α signaling pathway showed that Met-DOX presented more sensitivity to apoptosis and OS. Our findings indicate that metformin is a promising tool in the prevention of chemoresistance in patients with breast cancer submitted to doxorubicin-based treatments.

CORRELATION OF FREE HEMOGLOBIN LEVEL AND PLASMA NITRIC OXIDE IN PACKED RED CELL DURING BLOOD BANK STORAGE PERIOD

Stored red blood cells undergo morphological and biochemical changes with increased storage time, commonly refferred as the storage lesion, reduced integrity of erythrocyte membrane, causing hemolysis and increased free plasma hemoglobin level. Nitric Oxide (NO) is an endogenous vasodilator with the major role in vascular and blood flow regulation. Accumulation of free hemoglobin (fHb) during storage RBC hemolysis is thought to be correlated with elevated NO consuming causes low NO level that significantly impair endothelial function. This study aim to was to analyze the correlation of free Hb and plasma nitric oxide level during PRC storage period in the blood bank. This research was an analytic study with a cross-sectional design to 14 unit PRC those were stored in Dr. M. Djamil Padang Hospital Blood Bank. The study was conducted from May 2016 to August 2017. The assay was performed for 28 days of storage with one week intervals. Free plasma hemoglobin and NO level were analyzed by cyanmethemoglobin and colorimetry method respectively. Spearman test was used to access the correlation between free Hb level and nitric oxide plasma with p&lt;0.05 means significant. Most donors were male (85.7%) with age range were 33(9) years. Free Hb and nitric oxide level were significantly differenced in each week storage periods (p&lt;0.05). Two parameters was significantly had moderate negative correlation during storage periods (r=-0.56; p=0.01).

Characterization of eNOS ser‐600 Phosphorylation: A Link to the Cell Cycle

Nitric oxide (NO) is a gaseous, free‐radical signaling molecule produced by nitric oxide synthases (NOS). NO has a variety of cellular and physiological functions ranging from maintaining cardiovascular homeostasis to neurotransmission. In low concentrations NO is deemed physiologically crucial and in high concentrations NO is understood to be pathophysiological. The role of NO in the cell cycle is less understood, it is likely cell type specific and concentration dependent, in many cases low levels of NO promote proliferation and prevent apoptosis; at high levels NO has been shown to arrest cells and cause apoptosis. Endothelial nitric oxide synthase (eNOS) is one of three isoforms in the NOS family, it is the most regulated of the isoforms. Regulations include protein‐protein interactions and posttranslational modifications. The most commonly studied and well understood posttranslational modification of eNOS is phosphorylation. This work builds on our laboratories discovery of the ser‐600 phosphorylation site in the auto‐inhibitory loop of eNOS. Our work confirms this site is modulated in cells and links it to the cell cycle using immunofluorescence, flow cytometry, western blot analysis, in vitro kinase assays, and in vitro eNOS activity assays.Support or Funding InformationThis work was supported by NIH grant # R15 GM110634‐01A1 to Dr. Carol Chrestensen.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Raffia palm (Raphia hookeri) wine extenuates redox imbalance and modulates activities of glycolytic and cholinergic enzymes in hyperglycemia-induced testicular injury in type 2 diabetic rats.

This study investigated the effects of Raffia palm wine (RPW) on redox imbalance, glycolytic and cholinergic enzymes, and ATPase activities in hyperglycemia-induced oxidative testicular injury. Type 2 diabetes (T2D) was induced in male albino rats (Sprague-Dawley) by first administering 10% fructose solution for 14 days, before injecting with an intraperitoneal injection (40mg/kg body weight) of streptozotocin. Raffia palm wine was administered to two diabetic groups at 150 and 300mg/kg body weight (bw), when untreated diabetic group was used as a negative control, and metformin-fed group was served as a positive control. The rats were sacrificed after 5 weeks of treatment, and testes were harvested. Treatment with RPW led to lower levels of nitric oxide, malondialdehyde and myeloperoxidase activity, with concomitant elevation of reduced glutathione level, superoxide dismutase, catalase and ATPase activities. Raffia palm wine also inhibited glycogen phosphorylase, glucose-6-phosphatase, fructose-1,6-bisphosphatase, and acetylcholinesterase, while restoring the altered testicular morphology to near-normal. The results of this study suggest the therapeutic potentials of RPW against the effects of T2D on testicular functions and morphology. PRACTICAL APPLICATIONS: Diabetes mellitus is one of the fastest growing global epidemy, with most developing countries being at the receiving end owing to the cost of treatment. Testicular damage has been recognized as one of the complications of diabetes, and if left untreated will lead to erectile dysfunction followed by infertility. Raffia palm wine is among the common natural beverage in West Africa, which is utilized for both social and medicinal purposes. In this study, for the first time we showed its ability to protect diabetes-induced testicular injury in type 2 diabetic rats. This may be of great benefit in managing diabetes associated erectly dysfunction and male infertility, as Raffia palm wine is readily available in all seasons. This study will also improve the medicinal use of this wine, which may also indirectly improve its commercial benefit.

Periodic Assessment Of Antenatal And Post Natal Serum Endothelin1 and Nitric Oxide Levels In Hypertensive Disorders Of Pregnancy

Introduction: Hypertensive Disorder of Pregnancy (HDP) is an independent risk factor of cardiovascular (CVS) disease with persistent endothelial dysfunction, that occurs after the initial insult during pregnancy and persists post partum postulated to be the pathophysiology. Endothelin-1 (ET-1), a potent vasoconstrictor, has been identified as a pivotal mediator in both essential hypertension and HDP. Disturbances in Nitric Oxide (NO) bioavailability found in endothelial dysfunction may increase susceptibility to cardiovascular diseases such as hypertension. Materials and Methods: Thirty six pregnant women at 30-36 weeks period of gestation from the following categories (i) pregnancy induced hypertension (PIH) (ii) chronic hypertension during pregnancy (CH) and (iii) normal pregnant women (Control). Blood pressure indices measurements and sample collection was done at antepartum (30-36 weeks), post partum (8 weeks and 12 weeks). Endothelin-1 was measured using the Human ET-1 (Endothelin-1) ELISA Kit and serum NO was measured using the Nitric Oxide (total) detection kit. Results: All blood pressure indices were significantly higher in HDP patients compared to control during antenatal and post partum periods. Serum ET-1 was significantly higher in patients with HDP compared to control during antenatal until 3 months post partum. This was accompanied by significantly lower levels of serum NO in HDP patients. Conclusion: ET-1 levels in patients with history of HDP irrespective of post partum blood pressure contributes to persistent endothelial dysfunction.