Nitric Oxide Activity Research Articles

Biochemical and molecular-physiological aspects of the nitric oxide action in the utera

Biochemical and molecular-physiological aspects of the nitric oxide action in the utera

Biosynthesis and characterization of zinc oxide nanoparticles from an estuarine-associated actinobacterium Streptomyces spp. and its biotherapeutic applications

The present study was conducted to biosynthesize zinc oxide nanoparticles (ZnONPs) by an estuarine associated actinobacterium Streptomyces spp., characterized its spectral features and also determined its biomedical properties. At first, ZnONPs were synthesized by actinobacterial-mediated reduction of zinc oxide metal and then, it was characterized by following standard methodologies. The biosynthesized ZnONPs were confirmed through the reduction of Zn ions with the strong intensity range (3eV) at zinc region and the absorption range of 363nm. The XRD pattern of biosynthesized ZnONPs showed six intensive peaks with 2θ values of 36.38-83.34º. This ZnONPs exhibited spherical in shape with the average size ranges in between 51.42 and 65.13nm. The antimicrobial efficacy of various concentrations of ZnONPs was determined against both bacterial and fungal pathogens and it revealed the highest growth inhibition against E. coli (22mm) at 20μl concentration with the respective MIC and MBC values of 0.625μg/ml and ≤ 2.5μg/ml. Similarly, the antifungal activity represented the maximum (20mm) zone of growth inhibition against Candida fructus at 20μl concentration. Furthermore, the in vitro antioxidant, anti-inflammatory and antibiofilm efficiency of ZnONPs were tested. The result inferred that the maximum in vitro total antioxidant activity (70.86%), DPPH (79.26%), nitric oxide (67.31%) and hydroxyl radical (76.83%) scavenging activities were noted at 125μg/ml concentration of ZnONPs with the respective half maximal inhibitory concentration (IC50) values of 81.69, 83.42, 76.09 and 82.46μg/ml. In in vitro anti-inflammatory activity study, the maximum concentration (500μg/ml) of ZnONPs showed the inhibition of 94.62% with the IC50 value of 146.70μg/ml. Finally, the ZnONPs effectively reduced the biofilm formation with the highest percentage (91.08%) of inhibition against E. coli at 75μg/ml concentration. Based on these results, it could be concluded that the actinobacterium Streptomyces spp., mediated biosynthesized ZnONPs has biomedical potential with antimicrobial, antioxidant, antiinflammatory and antibiofilm properties.

Effects of Nitric Oxide Application on Antioxidant Enzyme Activities of Pepper Plants under Drought Stress

The purpose of the study was to determine the relationship between the messenger molecule Nitric oxide (NO) and antioxidative enzyme (SOD: Superoxide Dismutase; CAT: Catalase; APX: Ascorbate Peroxidase) activities in some metabolic changes that occur under the effect of drought stress in plants, to determine the possible roles of Nitric Oxide and to obtain complementary information. The experiment conducted in a controlled environment, and plant were cultured in containers containing Hoagland nutrient solution. For drought stress application, 10% Polyethylene Glycol (PEG 6000) was added to the nutrient solution, which is equivalent to -0.40 MPa osmotic potential. Before the drought stress is applied, pepper seedlings of Demre cv were pre-treated with different doses of Sodium Nitroprusside (SNP) and Carboxy-PTIO (potassium salt) (cPTIO) (SNP 0.01, SNP 1, SNP 100 and SNP 0.01 + cPTIO, SNP + cPTIO, SNP 100+ cPTIO). On the 10th day of the drought application, the growth parameters of the plants; the plant fresh weights and their Antioxidative Enzyme Activities (SOD, CAT, APX) were determined. In terms of plant growth parameters, both plant growth and antioxidant anzyme activities of plants pretreated with 0.01 and 1 doses of SNP were lower than the high dose of SNP and the PEG application without pretreatment. The reason for the low enzyme activities in these applications can be attributed to factors such as the excess accumulation of organic acids such as proline in the cells of the plants and the decrease in H2O2 and O-2 levels in the presence of SNP.

Synthesis and Characterization of Graphene Oxide Coated Au Nano Particles and The Study of its Application on Electro Catalytic Activity of Nitric Oxide

Synthesis and Characterization of Graphene Oxide Coated Au Nano Particles and The Study of its Application on Electro Catalytic Activity of Nitric Oxide

Simultaneous detection of melatonin and six metabolites of L-tryptophan pathway in rat gastric mucosa.

Melatonin (N-acetyl-5-methoxytryptamine) is an indoleamine synthesized in vertebrates mainly in the pineal gland, and is known to be involved mainly in thermoregulation and control of the circadian rhythm. That indoleamine can affect the auto-, para- and endocrine pathways, regulating body functions and affecting the metabolism of animals and humans. In addition to the pineal gland, melatonin can be synthesized in many extra-pineal tissues, mainly in the gastrointestinal tract. Previous studies have shown that melatonin plays an important role in the defense system of the gastrointestinal mucosa, demonstrating a protective effect on the gastrointestinal tract and the acceleration of healing of chronic ulcers through the scavenging of reactive oxygen metabolites (ROS) and the activation of protective nitric oxide (NO) and vasodilator neuropeptides released from the sensory afferent neurons. The process of converting the melatonin precursor L-tryptophan into melatonin is already known, but not all aspects of this process for the synthesis of other metabolites of this pathway have been fully elucidated and this issue remains poorly understood. In this study, the conversion of L-tryptophan to melatonin and other metabolites was determined in gastric mucosa collected from rats with or without intragastric (i.g.) melatonin or L-tryptophan administration, both administered at a single dose of 50 mg/kg. For the determination of five metabolites of L-tryptophan: kynurenine, 5-hydroxytryptamine, 5-hydroxytryptophan, anthranilic acid, indole-3-acetic acid together with melatonin, we have modified the previously developed high-performance liquid chromatography (HPLC) method using a native fluorescence detection system and UV-VIS. The obtained results show that: 1) L-tryptophan is converted into melatonin in the gastric mucosa during the day, e.g. after eating a meal containing L-tryptophan, as it was imitated and confirmed by our study, in which this amino acid was administered directly to the stomach, 2) the gastric mucosa is capable of producing melatonin in much greater amounts than those recorded in the blood serum of rats given a single dose of L-tryptophan, and 3) apart from melatonin, the only serum levels of these five metabolites of the L-tryptophan metabolic pathway are detectable, while their level in the gastric mucosa is low and barely detectable under physiological conditions. Our present observations support the notion that the gastric mucosa is one of the main sources of melatonin production from L-tryptophan outside the pineal gland.

Effect of glutathione S-transferase gene polymorphisms on semen quality in patients with idiopathic male infertility.

ObjectiveTo investigate the relationship between glutathione S-transferase enzyme (GSTM1, T1, and P1) genetic variants and semen quality in men with idiopathic infertility.MethodsSperm characteristics were measured using computer-assisted sperm analysis. The malondialdehyde (MDA), nitric oxide (NO), and total antioxidant capacity (TAC) activities were detected by spectroscopic analysis, and 8-hydroxy-2′-deoxyguanosine (8-OHdG) was detected by enzyme-linked immunosorbent assay.ResultsThis study included 246 idiopathic infertile men and 117 controls. The GSTM1(−), T1(−), and M1/T1(−/−) genotype frequencies significantly differed between the groups. The GSTM1(−) and T1(−) genotypes in idiopathic infertile men negatively correlated with sperm concentration, motility, mitochondrial membrane potential, and other parameters. However, these genotypes positively correlated with the amplitude of the lateral head displacement and NO and 8-OHdG levels. The GSTT1(−) genotype positively correlated with mean angular displacement and MDA activity. GSTM1(−) and T1(−) had a synergistic effect on semen quality. Sperm motility, normal morphology, straightness, and TAC were lower and amplitude of lateral head displacement and MDA were higher in the GSTP1(A/G + G/G) group than in the GSTP1(A/A) group among men with idiopathic infertility.ConclusionsGSTM1, T1, and P1 genetic variants may be risk factors for infertility by affecting the semen quality men with idiopathic oligoasthenospermia.

Lyophilized cell-free supernatants of Lactobacillus isolates exhibited antibiofilm, antioxidant, and reduces nitric oxide activity in lipopolysaccharide-stimulated RAW 264.7 cells.

BackgroundProbiotics can release bioactive substances known as postbiotics, which can inhibit pathogenic microorganisms, improve immunomodulation, reduce antioxidant production, and modulate the gut microbiota.MethodsIn this study, we evaluated the in vitro antimicrobial effects, antioxidant activity, and anti-inflammatory potential of 10 lyophilized cell-free supernatants (LCFS) of Lactobacillus isolates. LCFS was obtained via centrifugation and subsequent lyophilization of the supernatant collected from the culture medium ofeach isolate. The antibacterial and antibiofilm activities of the LCFS were determined using broth microdilution. The antioxidant potential was evaluated by measuring the total phenolic and flavonoid contents and 2,2-Diphennyl-1-picrylhydrazyl (DPPH) and 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+) radical scavenging activities.ResultsAll the isolates were able to inhibit the four tested pathogens. The isolates exhibited strong antibiofilm activity and eradicated the biofilms formed by Acinetobacter buamannii and Escherichia coli. All the prepared Lactobacillus LCFS contained phenols and flavonoids and exhibited antioxidant activities in the DPPH and ABTS+ radical scavenging assays. The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that LCFS was not cytotoxic to RAW 264.7 cells. In addition, the ten Lactobacillus LCFS decreased the production of nitric oxide.ConclusionsAll the isolates have beneficial properties. This research sheds light on the role of postbiotics in functional fermented foods and pharmaceutical products. Further research to elucidate the precise molecular mechanisms of action of probiotics is warranted.

Plasma proteome of brain-dead organ donors predicts heart transplant outcome

The pathophysiological changes related to brain death may affect the quality of the transplanted organs and expose the recipients to risks. We probed systemic changes reflected in donor plasma proteome and investigated their relationship to heart transplant outcomes. Plasma samples from brain-dead multi-organ donors were analyzed by label-free protein quantification using high-definition mass spectrometry. Unsupervised and supervised statistical models were used to determine proteome differences between brain-dead donors and healthy controls. Proteome variation and the corresponding biological pathways were analyzed and correlated with transplant outcomes. Statistical models revealed that donors had a unique but heterogeneous plasma proteome with 237 of 463 proteins being changed compared to controls. Pathway analysis showed that coagulation, gluconeogenesis, and glycolysis pathways were upregulated in donors, while complement, LXR/RXR activation, and production of nitric oxide and reactive oxygen species in macrophages pathways were downregulated. In point-biserial correlation analysis, lysine-specific demethylase 3A was moderately correlated with any grade and severe PGD. In univariate and multivariate Cox regression analyses myosin Va and proteasome activator complex subunit 2 were significantly associated with the development of acute rejections with hemodynamic compromise within 30 days. Finally, we found that elevated levels of lysine-specific demethylase 3A and moesin were identified as predictors for graft-related 1-year mortality in univariate analysis. We show that brain death significantly changed plasma proteome signature Donor plasma protein changes related to endothelial cell and cardiomyocyte function, inflammation, and vascular growth and arteriogenesis could predict transplant outcome suggesting a role in donor evaluation.

Nitrosylation of tissue transglutaminase enhances fibroblast migration and regulates MMP activation

In wound healing, the TG2 enzyme plays a dual functional role. TG2 has been shown to regulate extracellular matrix (ECM) stabilization by its transamidase activity while increasing cell migration by acting as a cell adhesion molecule. In this process, nitric oxide (NO) plays a particularly important role by nitrosylation of free cysteine ​​residues on TG2, leading to the irreversible inactivation of the catalytic activity.In this study, transfected fibroblasts expressing TG2 under the control of the tetracycline-off promoter were treated with NO donor S-nitroso-N-acetyl penicillamine (SNAP) to analyze the interplay between NO and TG2 in the regulation of cell migration/invasion as well as TGF-β1-dependent MMP activation. Our results demonstrated that inhibition of TG2 cross-linking activity by SNAP promoted the migration and invasion capacity of fibroblasts by hindering TG2-mediated TGF-β1 activation. While the inhibition of TG2 activity by NO downregulated the biosynthesis and activity of MMP-2 and MMP-9, that of MMP-1a and MMP-13 was shown to be upregulated in a TGF-β1-dependent manner under the same conditions. In the presence of SNAP, interaction of TG2 with its cell surface binding partners Integrin-β1 and Syndecan-4 was reduced, which was paralleled by an increase in TG2 and PDGF association. These findings suggests that migratory phenotype of fibroblasts can be regulated by the interplay between nitric oxide and TG2 activity.

Phytochemical Screening and Therapeutic Effects of Binahong (Anredera cordifolia (Ten.) Steenis) Leaves

In this review, the function of secondary metabolites in Binahong (Anredera cordifolia (Ten.) Steenis) leaves were summarized for their therapeutic effects such as antimicrobial activity, anti-inflammatory, wound healing, lowering the blood pressure, and antihyperlipidemic activity as a potential medicinal plant to treat disease. The phytochemical screening of Binahong leaves extracted by using particular solvent showed positive results to the presence of phenolic, steroid, terpenoid, alkaloid, and saponin compounds contained in the extract. The antimicrobial activity is able to disrupt cell membrane activity and inhibit bacterial growth, meanwhile anti-inflammatory ability is able to decrease anti-inflammatory agents level. It also decreases blood pressure by regulation of lipid metabolites, reduction of peripheral resistance, and upregulation of nitric oxide activity. The ethanolic extract of Binahong leaves can reduce fat deposits or layers in the endothelial cells, cholesterol, triglycerides, and low-density lipoprotein (LDL) cholesterol levels, and decrease the levels of malondialdehyde (MDA formation) from the lipid peroxidation to prevent hyperlipidemia and heart disease.

Cellgevity® attenuates liver distruption, oxidative stress and inflammation in STZ-diabetic male rats

The imbalance between reactive oxygen species (ROS) production and the innate antioxidant defence promotes oxidative distress and contributes to tissue dysfunctions which are the hallmark of diabetic complications. Diabetic hepatic injury is less reported, yet it is a target organ damage condition related to diabetes. This study investigated the hepato-protective potentials of Cellgevity® in Streptozotocin (STZ)-diabetic male rats. The rats were assigned into – Control group and diabetes induced groups (DM-untreated, DM-treated 1 and DM-treated 2 groups respectively). Separate treatment of the DM-treated 1 and DM-treated 2 groups with therapeutic doses of Cellgevity® (25 mg/kg and 40 mg/kg BW respectively) was conducted for 30 days, while the control and DM-untreated groups receieved oral distilled water (placebo). The animals were then sacrificed and their sera were evaluated for total antioxidant status and biomarkers of hepatic function. The rats’ liver homogenates were also evaluated for inflammation, nitric oxide, lipid peroxidation and antioxidant enzymes activity. Treatment with Cellgevity® significantly increased the serum total antioxidant status (2 fold) and it reduced the serum ALT, AST, ALP and bilirubin levels compared to untreated diabetes by at least 1.4 fold. Hepatic activities of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) were significantly increased by over 130%, while liver TNF-α, nitrite and MDA levels were reduced by over 140% compared to the diabetic untreated group. The results present promising evidence for the therapeutic potential of Cellgevity® against diabetes-induced liver dysfunction, which might be through the modulation of ROS and inflammation production respectively.

Synergistic effect between quinpirole and L-NAME as well as sulpiride and L-arginine on the modulation of anxiety and memory processes in the 6-OHDA mouse model of Parkinson's disease: An isobologram analysis

We evaluated interactions between dopamine D2 receptor and nitric oxide (NO) actions on the regulation of anxiety and memory in the 6-hydroxydopamine (6-OHDA) mouse model of Parkinson's disease (PD). A unilateral guide cannula was stereotaxically implanted over the right striatum. Elevated plus-maze test (EPM) test–retest protocol was employed to evaluate anxiety and memory in mice. The results revealed that injection of L-NAME (9 mg/kg) induced anxiolytic and amnesic effects, while L-arginine (9 mg/kg) produced anxiogenic and memory-improvement effects in the 6-OHDA mouse model of PD. Administration of sulpiride (20 mg/kg) induced anxiogenic and memory-improvement effects, whereas quinpirole (20 mg/kg) caused anxiolytic and amnesic effects in PD mice. Co-injection of sulpiride (5, 10, and 20 mg/kg) plus L-NAME (3 mg/kg) induced anxiolytic and amnesic effects. Co-injection of sulpiride (20 mg/kg) plus L-arginine (3 mg/kg) induced anxiogenic and memory-improvement effects. Co-administrations of quinpirole (20 mg/kg) and L-NAME (3 mg/kg) induced anxiolytic effect, but co-administration of quinpirole (20 mg/kg) plus L-arginine (3 mg/kg) caused anxiogenic and memory-improvement effects. The isobologram analysis revealed that there is a synergistic effect between sulpiride and L-arginine as well as quinpirole and L-NAME upon induction of anxiogenic and anxiolytic effects, respectively in PD mice. Our results suggested: (1) NO and dopamine D2 receptor mechanisms affect anxiety and memory in PD mice; (2) L-NAME reversed anxiogenic and memory-improvement effect induced by sulpiride; (3) Anxiolytic and amnesic effects induced by quinpirole reversed by L-arginine; (4) There is a synergistic effect between dopamine D2 receptor and NO systems on the modulation of anxiety and memory.

Antioxidant activity, xanthine oxidase inhibition and acute oral toxicity of Dillenia philippinensis Rolfe (Dilleniaceae) leaf extract

Context: Hyperuricemia is a metabolic syndrome characterized by a high serum uric acid level with an increased risk of gout, diabetes, cardiovascular and renal disease. Allopurinol is a widely known xanthine oxidase inhibitor that lowers serum uric acid production. Documented adverse effects of the drug raise the need for a natural alternative for xanthine oxidase inhibitor that is safe and effective. This research is driven to address the need by capitalizing on previous studies made on the Dillenia species showing nephroprotective and antihyperuricemic activity. Aims: To evaluate the antioxidant capacity, xanthine oxidase inhibitory activity, and acute toxicity of Dillenia philippinensis; a plant locally known as Katmon and endemic to the Philippines. Methods: The D. philippinensis ethanolic leaf extract (DPELE) was partitioned using solvents of different polarities, hexane, ethyl acetate, and butanol. The antioxidant activity of the DPELE and the sub-extracts was assessed using nitric oxide, hydrogen peroxide, and hydroxyl radical scavenging assays against ascorbic acid, while the xanthine oxidase inhibitory activity was examined against allopurinol as a standard. The acute toxicity was performed using Sprague-Dawley rats following OECD 425 guidelines. Results: The D. philippinensis ethyl acetate fraction (DPEAF) had the highest scavenging activity of nitric oxide, hydrogen peroxide, and hydroxyl (IC50 = 210.00, 70.92 and 59.88 µg/mL) free radicals, respectively. Also, the xanthine oxidase dose-dependent inhibitory activity was observed to be highest in the DPEAF with an IC50 = 23.09 µg/mL. Lastly, neither toxicity nor mortalities were observed in rats treated with the DPELE and the DPEAF for 14 days. The approximate lethal dose for DPELE and DPEAF was higher than 2000 mg/kg BW. Conclusions: This study shows that the DPELE and DPEAF are potential sources of natural antioxidants and xanthine oxidase inhibitors, which may be used in the treatment of hyperuricemia and are presumed safe to be used orally.

Nitric Oxide Mediation in Hydroxyurea and Nitric Oxide Metabolites' Inhibition of Erythroid Progenitor Growth.

In several systems, hydroxyurea has been shown to trigger nitric oxide (NO) release or activation of NO synthase (NOS). To elucidate this duality in its pharmacological effects, during myelosuppression, we individually examined hydroxyurea’s (NO releasing agent) and NO metabolites’ (stable NO degradation products) effects on erythroid colony growth and NOS/NO levels in mice using NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO). Hydroxyurea and nitrite/nitrate decreased the bone marrow cellularity that was blocked by PTIO only for the NO metabolites. Hydroxyurea inhibition of colony-forming unit-erythroid (CFU-E) formation and reticulocytes was reversed by PTIO. Moreover, hydroxyurea, through a negative feedback mechanism, reduced inducible NOS (iNOS) expressing cells in CFU-E, also prevented by PTIO. Nitrate inhibition of burst-forming units-erythroid (BFU-E) colony growth was blocked by PTIO, but not in mature CFU-E. The presented results reveal that NO release and/or production mediates the hydroxyurea inhibition of mature erythroid colony growth and the frequency of iNOS immunoreactive CFU-E.

Photobiomodulation (λ=808nm) and Platelet-Rich Plasma (PRP) for the Treatment of Acute Rheumatoid Arthritis in Wistar Rats.

Introduction: Rheumatoid arthritis (RA) causes inflammation, pain, edema, and articular degradation and its treatment can be based on anti-inflammatory drugs, photobiomodulation (PBM) and/or platelet-rich plasma (PRP) that can decrease cell flow and promote local healing. In the present study, we evaluate the effects of PBM and PRP on acute arthritis in Wistar rats through inflammatory and oxidative stress parameters. Methods: Thirty female Wistar rats were assigned to five groups (n=6, each group): Control, Sham, PRP, Laser, and PRP+Laser. For arthritis induction, all animals of groups Sham, PRP, Laser and PRP+Laser received an intraarticular injection of Zymosan® (200µg) in the right knee. Twenty-four hours post-arthritis induction, PRP was prepared and injected (8 × 105 of platelets) in animals of PRP and PRP+Laser groups. PBM was performed in Laser and PRP+Laser groups by single-dose therapy with the GaAlAs laser (λ=808 nm, P=25 mW, fluence=30 J/cm2, beam area=0.02 mm2, t=33 seconds, E=0.825 J, punctual application). After seven days of induction, serum samples were collected and thiobarbituric acid reactive substances (TBARS), nitric oxide (NO) and catalase activity were analysed. Morphological parameters were measured for inflammation areas, cartilage thickness, and C3 protein expression in knee samples. Statistical analysis was performed with an ANOVA test and Tukey's post-hoc test with a significance level of 5% (P<0.05). Results: NO was lower in the treated groups compared to the Sham group, and TBARS did not show any differences, while catalase showed greater activity between PRP+Laser versus PRP (P<0.05). Inflammatory areas and cartilage thickness were lower in the treated groups compared to Sham (P<0.05), while no differences in C3 protein expression was observed. Conclusion: PBM associated with PRP is better for anti-inflammatory and joint preservation by morphological aspects and NO levels that concern a potential clinical application.

Three-Dimensional RAW264.7 Cell Model on Electrohydrodynamic Printed Poly(ε-Caprolactone) Scaffolds for In Vitro Study of Anti-Inflammatory Compounds

Inflammation plays an essential role in the human immune system, and anti-inflammatory compounds are important to promote health. However, the in vitro screening of these compounds is largely dependent on flat biology. Herein, we report our efforts in establishing a 3D inflammation murine macrophage model. Murine macrophage RAW 264.7 cells were cultured on poly(ε-caprolactone) (PCL) scaffolds fabricated through an electrohydrodynamic jetting 3D printer and their behavior were examined. Cells on PCL scaffolds showed a 3D shape and morphology with multilayers and a lower proliferation rate. Moreover, macrophages were not activated by scaffold material PCL and 3D microenvironment. The 3D cells showed greater sensitivity to lipopolysaccharide stimulation with higher production activity of nitric oxide (NO), nitric oxide synthases (iNOS), and cyclooxygenase-2 (COX-2). Additionally, the 3D macrophage model showed lower drug sensitivity to commercial anti-inflammatory drugs including aspirin, ibuprofen, and dexamethasone, and natural flavones apigenin and luteolin with higher IC50 for NO production and lower iNOS and COX-2 inhibition efficacy. Overall, the 3D macrophage model showed promise for higher accurate screening of anti-inflammatory compounds. We developed, for the first time, a 3D macrophage model based on a 3D-printed PCL scaffold that provides an extracellular matrix environment for cells to grow in the 3D dimension. 3D-grown RAW 264.7 cells showed different sensitivities and responses to anti-inflammatory compounds from its 2D model. The 3D cells have lower sensitivity to both commercial and natural anti-inflammatory compounds. Consequently, our 3D macrophage model could be applied to screen anti-inflammatory compounds more accurately and thus holds great potential in next-generation drug screening applications.

Prunetin inhibits nitric oxide activity and induces apoptosis in urinary bladder cancer cells via CASP3 and TNF-α genes.

Urinary bladder cancer (UBC) is considered one of the most prevalent malignant tumors worldwide. Complementary and integrative approaches for the treatment of bladder cancer, such as the intake of isoflavonoid phytoestrogens, are of increasing interest due to the risk of mortality and long-term morbidity associated with surgical procedures. The biological effects of prunetin, one of the less-studied phytoestrogens, have not yet been examined in this respect. Therefore, this study aimed to explore the efficacy of prunetin on UBC cells (RT-4). METHODS ANDRESULTS: The cytotoxicity and nitric oxide synthase activities of prunetin were determined in cell cultures. The expression of apoptosis-related genes was determined with RT-PCR. Cell cycle assays were performed using a flow cytometer and cellular apoptotic rate was measured. The results suggested that prunetin has cytotoxic effects at 21.11 µg/mL on RT-4 cells. Flow cytometry analysis showed that prunetin induced apoptosis and arrested th cell cycle in the G0/G1 phase. Prunetin exposure was associated with increases in CASP3 and TNF-α gene expression in RT-4 cells at doses of 21.11 and 42.22 µg/mL, respectively. Strong nitric oxide inhibition was observed at IC50 of 5.18 µg/mL under macrophage mediated inflammatory circumstances. Prunetin possesses anti-cancer properties and may be a candidate compound for the prevention of UBC. This is the first study that evaluated prunetin for its in vitro antitumor activities, clarified its possible apoptotic molecular mechanism and provided novel insights into its anti-inflammatory nature and effects on the expression of related key genes.

Screening of QTc interval and global autonomic activity in autosomal dominant polycystic kidney disease and atherosclerotic renal artery stenosis hypertensive patients.

Arterial hypertension (AH) represents a major risk factor for cardiovascular disease and is associated to several complications, such as prolonged corrected QT (QTc) interval and impaired heart rate variability (HRV). Secondary causes of AH include autosomal dominant polycystic kidney disease (ADPKD) and atherosclerotic renal artery stenosis (ARAS), both known to be related to arrhythmic risk and autonomic imbalance. The aim of the study is to evaluate whether global autonomic activity and QTc interval differently affect ADPKD and ARAS hypertensive patients. An observational study was performed on 59 patients: 16 ADPKD patients and 19 diagnosed with ARAS, compared to 24 healthy controls (HC). All patients underwent clinical evaluation, biochemical lab tests, 24-hour electrocardiogram (ECG) and renal Doppler ultrasound. HRV was assessed through the analysis of 24-hour ECG to detect standard deviation of normal-to-normal RR intervals (SDNN). QTc interval was defined as prolonged when > 440 msec. SDNN was significantly lower in ADPKD and ARAS patients than HC (p < 0.0001) and no significant differences were found between ADPKD and ARAS patients (p > 0.05). QTc was found significantly higher in ARAS patients than HC (p = 0.001) and in ARAS patients than ADPKD patients (p = 0.004). The pathogenesis of hypertension in ADPKD and ARAS patients is related to the activation of the renin angiotensin aldosterone system (RAAS). In ADPKD, cyst enlargement leads to kidney ischemia and renin release, associated to endothelial dysfunction, low nitric oxide and sympathetic tone activation. Differently, reduction in renal perfusion pressure activates RAAS and renal adrenergic nerves in ARAS patients. We can speculate that prolonged QTc interval is more present in ARAS vs. ADPKD hypertensive patients due to a greater activation of RAAS. We suggest adding 24-hour HRV evaluation in association with traditional risk factors in course of ADPKD and ARAS hypertension to better stratify cardiovascular risk in these groups of patients.

Blood circulation activating effect of Sanqi (Radix Notoginseng) on venous thromboembolism rat.

To explore the blood circulation activating effect and mechanism of Sanqi (Radix Notoginseng) in vivo, using a venous thromboembolism (VTE) rat model. We established the VTE rat model, and then intervened with low molecular weight heparin (LMWH), as well as low, medium and high doses of Sanqi (Radix Notoginseng), to observe the blood circulation activating effect of Sanqi (Radix Notoginseng) on VTE rats. After the treatment with high concentrations of Sanqi (Radix Notoginseng), the pulmonary thromboembolism was alleviated, and the lower limb thrombosis was markedly improved. Moreover, the expression quantities of plasma activated partial thromboplastin time, prothrombin time and D-dimer, as well as endothelin, von Willebrand factor, and plasminogen activator inhibitor-1 in thrombosis segment tissues were markedly down-regulated; while those of nitric oxide and tissue-type plasminogen activator were up-regulated. After low and medium concentration Sanqi (Radix Notoginseng) treatment, no obvious improvement was observed in each index. Moreover, the high concentration Sanqi (Radix Notoginseng) showed comparable efficacy to the positive drug LMWH. This data suggests that high concentration of Sanqi (Radix Notoginseng) is effective in preventing and treating VTE.

Is Basal Nitric Oxide Activity of the Renal Vasculature Altered? Analysis of a Randomized Controlled Trial Comparing Two Combination Therapies

Is Basal Nitric Oxide Activity of the Renal Vasculature Altered? Analysis of a Randomized Controlled Trial Comparing Two Combination Therapies