Overproduction Of Nitric Oxide Research Articles

Anti-inflammatory activity of flavonols via inhibiting MAPK and NF-κB signaling pathways in RAW264.7 macrophages

Fisetin (Fis), quercetin (Que), and myricetin (Myr) are flavonols with similar structure but different number of hydroxyl groups. The present research focused on the anti-inflammatory effect of these three flavonols in lipopolysaccharide-stimulated RAW264.7 cells. The number and site of hydroxyl group in flavonols obviously affected their anti-inflammation activity. These flavonols suppressed the overproduction of nitric oxide. Fis showed the best activity with an inhibition rate of 52% at 20 μM. Moreover, the flavonols reduced the levels of ROS, TNF-α, and IL-6. The mechanistic study showed that they inhibited the activation of NF-κB and MAPK pathways by suppressing the phosphorylation of IκBα, p65, JNK, ERK, p38, MEK, and reducing the nuclear translocation of NF-κB p65. In addition, the metabolism of the flavonols was examined. The results indicated that Fis was both methylated and glucuronidated. Que and Myr were mainly transformed into methylated products. This study highlights the anti-inflammatory activity of flavonols, particularly Fis, which has the potential for the prevention or treatment of inflammation as an adjuvant medicine or food additive.

Association between intestinal lymphangiectasia and expression of inducible nitric oxide synthase in dogs with lymphoplasmacytic enteritis.

Intestinal lymphangiectasia (IL) is a common complication in dogs. Since nitric oxide (NO) is known to relax the lymphatic vessel, we evaluated inducible NO synthase (iNOS) expression using immunohistochemistry in 13 dogs with lymphoplasmacytic enteritis (LPE) with or without IL. The duodenal iNOS expressing cells were significantly increased in dogs with IL-negative or IL-positive LPE dogs (P=0.025, P=0.007) compared with control dogs. However, there was no significant difference in iNOS expression between IL-positive and IL-negative tissues. Based on these results, there is no clear evidence for the NO overproduction in the pathogenesis of IL in dogs with LPE. Factors other than NO could, thus, contribute to IL in dogs with LPE.

Bioactive Diterpenes, Norditerpenes, and Sesquiterpenes from a Formosan Soft Coral Cespitularia sp.

Chemical investigation of the soft coral Cespitularia sp. led to the discovery of twelve new verticillane-type diterpenes and norditerpenes: cespitulins H–O (1–8), one cyclic diterpenoidal amide cespitulactam L (9), norditerpenes cespitulin P (10), cespitulins Q and R (11 and 12), four new sesquiterpenes: cespilins A–C (13–15) and cespitulolide (16), along with twelve known metabolites. The structures of these metabolites were established by extensive spectroscopic analyses, including 2D NMR experiments. Anti-inflammatory effects of the isolated compounds were studied by evaluating the suppression of pro-inflammatory protein tumor necrosis factor-α (TNF-α) and nitric oxide (NO) overproduction, and the inhibition of the gene expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in lipopolysaccharide-induced dendritic cells. A number of these metabolites were found to exhibit promising anti-inflammatory activities.

S2813 Pulmonary Hypertension as a Rare Complication After Orthotopic Liver Transplant in a Patient With NASH Cirrhosis Complicated by Hepatopulmonary Syndrome

Introduction: Hepatopulmonary syndrome (HPS) is a phenomenon that results in an intrapulmonary shunt leading to dyspnea and hypoxemia with poor response to oxygen supplementation. In this case, we describe a rare occurrence of pulmonary hypertension developing after orthotopic liver transplant (OLT) in a cirrhotic patient with HPS. Case Description/Methods: 61-year-old female with a history of NASH cirrhosis complicated by esophageal varices and hepatic encephalopathy underwent liver transplant for worsening shortness of breath secondary to HPS. Two years after transplant, she presented with worsening dyspnea, saturating 86% on room air. Chest X ray and CT scan showed enlarged pulmonary trunk without consolidation or pulmonary edema, and 2D-echo showed normal LV ejection fraction with no valvular abnormality. Right and left heart catheterization showed high mPAP of 66 mm Hg consistent with severe pulmonary hypertension, with normal wedge pressure. She was started on intravenous epoprostenol and riociguat for pulmonary arterial hypertension, as well as diuretics. Repeat right heart catheterization during hospitalization showed mild improvement of mPAP to 57 mm Hg. However, patient’s hospital course was complicated by worsening shortness of breath requiring intubation, acute kidney injury and blood loss anemia from variceal bleed. Unfortunately, the patient’s condition continued to deteriorate, and comfort measures were initiated. Discussion: Hepatopulmonary syndrome, prevalent in 5-23% of cirrhotic patients, classically presents with progressive dyspnea and exercise induced hypoxemia. Mechanism consists of pulmonary vascular dilation secondary to overproduction of nitric oxide affectively creates an intrapulmonary shunt causing hypoxemia. HPS is reversible with liver transplantation, however, some transplants result in subsequent pulmonary hypertension. Postulated mechanisms include unmasking of underlying pulmonary hypertension with OLT, and increased PVR due to increased blood flow in HPS. In a case series by Zopey et al., three patients achieved spontaneous resolution of HPS and subsequently developed portopulmonary hypertension before liver transplantation, highlighting the intricate balance between pulmonary vasodilators and vasoconstrictors. Our case adds to the growing recognition of the development of pulmonary hypertension after liver transplant in patients with HPS. It is important for providers to be aware of this phenomenon as mortality is estimated to be 69%.

Light-activated nitric-oxide overproduction theranostic nanoplatform based on long-circulating biomimetic nanoerythrocyte for enhanced cancer gas therapy

Light-activated nitric-oxide overproduction theranostic nanoplatform based on long-circulating biomimetic nanoerythrocyte for enhanced cancer gas therapy

Nitric Oxide Impacts Human Gut Microbiota Diversity and Functionalities

ABSTRACTThe disruption of gut microbiota homeostasis has been associated with numerous diseases and with a disproportionate inflammatory response, including overproduction of nitric oxide (NO) in the intestinal lumen. However, the influence of NO on the human gut microbiota has not been well characterized yet. We used in vitro fermentation systems inoculated with human fecal samples to monitor the effect of repetitive NO pulses on the gut microbiota. NO exposure increased the redox potential and modified the fermentation profile and gas production. The overall metabolome was modified, reflecting less strict anaerobic conditions and shifts in amino acid and nitrogen metabolism. NO exposure led to a microbial shift in diversity with a decrease in Clostridium leptum group and Faecalibacterium prausnitzii biomass and an increased abundance of the Dialister genus. Escherichia coli, Enterococcus faecalis, and Proteus mirabilis operational taxonomic unit abundance increased, and strains from those species isolated after NO stress showed resistance to high NO concentrations. As a whole, NO quickly changed microbial fermentations, functions, and composition in a pulse- and dose-dependent manner. NO could shift, over time, the trophic chain to conditions that are unfavorable for strict anaerobic microbial processes, implying that a prolonged or uncontrolled inflammation has detrimental and irreversible consequences on the human microbiome.IMPORTANCE Gut microbiota dysbiosis has been associated with inflammatory diseases. The human inflammatory response leads to an overproduction of nitric oxide (NO) in the gut. However, so far, the influence of NO on the human gut microbiota has not been characterized. In this study, we used in vitro fermentation systems with human fecal samples to understand the effect of NO on the microbiota: NO modified the microbial composition and its functionality. High NO concentration depleted the microbiota of beneficial butyrate-producing species and favored potentially deleterious species (E. coli, E. faecalis, and P. mirabilis), which we showed can sustain high NO concentrations. Our work shows that NO may participate in the vicious circle of inflammation, leading to detrimental and irreversible consequences on human health.

Hostaflavone A from Hosta plantaginea (Lam.) Asch. blocked NF-κB/iNOS/COX-2/MAPKs/Akt signaling pathways in LPS-induced RAW 264.7 macrophages

Ethnopharmacological relevanceHostaflavone A (HA) is a new flavonoid component isolated from the flower of Hosta plantaginea (Lam.) Asch., which is commonly used as a folk herbal to treat inflammatory diseases in China. Nevertheless, the anti-inflammatory effect of HA remains unknown. Aim of the studyThis work aimed to evaluate the HA with anti-inflammatory activity and mechanism in RAW 264.7 macrophages activated by lipopolysaccharide (LPS). Materials and methodsAnti-inflammatory effect of HA was evaluated by measuring of cell viability, nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β) and IL-6 levels in RAW 264.7 cells. In parallel, the HA action mechanism of nuclear factor kappa B (NF-κB) p65, inhibitor of NF-κB (IκB), inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinase (Erk), p38, and protein kinase B (Akt) were detected by Western blot analysis. ResultsHA has no cytotoxicity at concentrations as high as 40 μM. Besides, HA concentration-dependently clearly suppressed the overproduction of NO, PGE2, TNF-α, IL-1β and IL-6 in RAW 264.7 cells induced by LPS. In addition, HA remarkably reduced the upregulation of phosphorylated NF-κB p65, phosphorylated IκB, phosphorylated JNK, phosphorylated Erk and phosphorylated p38, together with iNOS and COX-2 protein expressions in a concentration-dependent manner. ConclusionHA blocked the LPS activated inflammation via suppressing NF-κB, iNOS, COX-2, mitogen-activated protein kinases (MAPKs) and Akt pathways in RAW 264.7 cells, and might be a new anti-inflammatory agent.

Plantanone C attenuates LPS-stimulated inflammation by inhibiting NF-κB/iNOS/COX-2/MAPKs/Akt pathways in RAW 264.7 macrophages

The flowers of Hosta plantaginea (Lam.) Aschers are commonly used for the treatment of inflammatory diseases in traditional Chinese medicine with limited scientific evidence. Plantanone C (PC) is a new phytochemical isolated from H. plantaginea flowers; nevertheless, the anti-inflammatory effect remains unknown. Herein, we aimed to study the anti-inflammatory effects of PC and its underlying molecular mechanisms in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The cell viability of PC-treated RAW 264.7 macrophage was measured by the Cell Counting kit-8 (CCK-8) assay. The anti-inflammatory effect of PC was investigated by measuring the levels of inflammatory mediators and pro-inflammatory cytokines using the Griess reaction and enzyme-linked immunosorbent assay (ELISA). Furthermore, the mechanism of action of PC was evaluated by Western blot analysis. The results showed that PC was not cytotoxic at concentrations as high as 40 μM. Furthermore, PC potently suppressed LPS-stimulated overproduction of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β) and IL-6 in RAW 264.7 macrophages. Western blot demonstrated that PC remarkably suppressed the phosphorylation of nuclear factor kappa-B (NF-κB) p65, inhibitor of NF-κB (IκB), c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinase (Erk), p38, and protein kinase B (Akt), as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) in a concentration-dependent manner. Taken together, these findings suggest that PC exhibits anti-inflammatory effects by inhibiting NF-κB, iNOS, COX-2, mitogen-activated protein kinases (MAPKs), and Akt signaling pathways in RAW 264.7 macrophages.

Nitrosative stress parameters and the level of oxidized DNA bases in patients with multiple sclerosis.

Multiple sclerosis (MS) is a neurodegenerative disease with various factors affecting its etiology. Overproduction of nitric oxide and subsequent lesions of biopolymers are some of the possible causes of the disease. This study aimed to measure the most relevant nitrosative and oxidative stress biomarkers and the level of modified DNA bases in patients with MS. Each parameter was assayed in 25 patients with MS and 25 healthy controls. This study involved detecting blood plasma and serum nitric oxide metabolites by chemiluminescence detector Sievers NOA-280i, malondialdehyde (MDA) measurements with thiobarbituric acid reactive substance (TBARS) assay, detection of oxidized purines and pyrimidines with the enzyme-modified comet assay. Statistical analysis of the results was performed by one-way analysis of variance (ANOVA) and unpaired t test for the comparison of less than three data sets. DNA single-strand breaks, levels of modified purines and pyrimidines, as well as nitrite and nitrate levels in plasma and serum samples, were significantly higher in patients with MS than in healthy controls. On the contrary, MDA levels appeared to be lower in patients with MS.

Glycogen-based pH and redox sensitive nanoparticles with ginsenoside Rh2 for effective treatment of ulcerative colitis

The purpose of this study is to construct a pH and redox sensitive nanoparticle to effectively deliver ginsenoside Rh2 for the treatment of ulcerative colitis (UC). Herein, glycogen was modified by urocanic acid and α-lipoic acid (α-LA) to obtain an amphiphilic polymer (LA-UaGly). Such polymer LA-UaGly could self-assemble to form nanoparticles (Blank NPs) in water with excellent stability, which could also successfully encapsulated ginsenoside Rh2 to form Rh2 nanoparticles (Rh2 NPs) with encapsulation efficiency of 74.36 ± 0.34%. DLS analysis indicated Rh2 NPs were spherical with a particle size of 128.9 ± 0.3 nm. As expected, Rh2 NPs exhibited typical pH and redox dual response release behaviour as well as the excellent in vivo safety. In vitro tests showed that Rh2 NPs could effectively internalize and release Rh2 into RAW264.7 cells, and protect cells from apoptosis (p < 0.05). More interestingly, Rh2 NPs exhibited strong anti-inflammatory activity via significantly inhibiting the overproduction of nitric oxide (NO) and inflammatory cytokines (TNF-α, IL-1β and IL-6) (p < 0.05). In vivo experiments suggested that Rh2 NPs significantly ameliorated the weight loss, colon length, disease activity index (DAI) score, and myeloperoxidase (MPO) activity in mice caused by dextran sulfate sodium salt (DSS) (p < 0.05). Simultaneously, pathological analysis proved that Rh2 NPs could significantly reduce histological damage and inflammatory infiltration in mice. Rh2 NPs could also effectively regulate the intestinal flora of mice by improving the species uniformity and abundance of the intestinal flora of mice and restoring the species diversity of the intestinal flora. In addition, both in vivo and in vitro experiments proved that Rh2 NPs had stronger anti-inflammatory activity than Rh2. This study provides a promising strategy for the effective treatment of UC.

Perspective of Immunopathogenesis and Immunotherapies for Kawasaki Disease.

Kawasaki Disease (KD) is an acute inflammatory illness that mostly occurs in children below 5 years of age, with intractable fever, mucocutaneous lesions, lymphadenopathy, and lesions of the coronary artery (CAL). KD is sharing clinical symptoms with systemic inflammatory syndrome in children (MIS-C) which is related to COVID-19. Certain genes are identified to be associated with KD, but the findings usually differ between countries and races. Human Leukocyte Antigen (HLA) allele types and toll-like receptor (TLR) expression are also correlated to KD. The acute hyperinflammation in KD is mediated by an imbalance between augmented T helper 17 (Th17)/Th1 responses with high levels of interleukin (IL)-6, IL-10, IL-17A, IFN-γ, and IP-10, in contrast to reduced Th2/Treg responses with lower IL-4, IL-5, FoxP3, and TGF-β expression. KD has varying phenotypic variations regarding age, gender, intravenous immunoglobulin (IVIG) resistance, macrophage activation and shock syndrome. The signs of macrophage activation syndrome (MAS) can be interpreted as hyperferritinemia and thrombocytopenia contradictory to thrombocytosis in typical KD; the signs of KD with shock syndrome (KDSS) can be interpreted as overproduction of nitric oxide (NO) and coagulopathy. For over five decades, IVIG and aspirin are the standard treatment for KD. However, some KD patients are refractory to IVIG required additional medications against inflammation. Further studies are proposed to delineate the immunopathogenesis of IVIG-resistance and KDSS, to identify high risk patients with genetic susceptibility, and to develop an ideal treatment regimen, such as by providing idiotypic immunoglobulins to curb cytokine storms, NO overproduction, and the epigenetic induction of Treg function.

Anti-allergic and anti-inflammatory effects of hydrosol extracted from Zanthoxylum schinifolium branch

Zanthoxylum schinifolium Sieb. et Zucc. (syn. Fagara schinifolia Engler) was studied for its potential anti-inflammatory properties. The hydrosol extract prepared from the Z. schinifolium branch was analyzed by gas chromatography/mass spectrometry. Here, five main chemical components were identified in the hydrosol of the branches of this shrub. The main chemical compounds in the branch inhibited both an Immunoglobulin E (IgE)-antigen complex and a dinitrophenyl-bovine serum albumin (DNP-BSA)-induced β-hexosaminidase release in a dose-dependent manner in RBL-2H3 mast cells, and at the tested concentrations did not show cytotoxicity to RBL-2H3 cells. Moreover, hydrosol obtained from the branch substantially inhibited a lipopolysaccharide (LPS) induced overproduction of intracellular active oxygen (ROS) and nitric oxide (NO). Consistently, the soluble N-ethylmaleimide-sensitive factor-attachment protein receptor (SNARE) proteins of SNAP23, syntaxin4, VAMP7, and VAMP8 were remarkably decreased through hydrosol treatment. Hydrosol suppressed the activation of SNARE proteins in DNP-BSA-stimulated RBL-2H3 cells and inhibited ROS and NO in LPS-stimulated RAW264.7 cells. Camphor and estragole are the main chemical components of hydrosol and downregulate the LPS-induced phosphorylation of the SNARE proteins. The hydrosol obtained from the branch of Z. schinifolium has therapeutic benefits for allergic inflammatory diseases.

Гепатопульмональный синдром. Обзор литературы

Liver cirrhosis is often accompanied by complications from the pulmonary system. These include hydrothorax, portopulmonary hypertension and hepatopulmonary syndrome. Hepatic hydrothorax affects about 6-10% of patients with end-stage disease, which results in the passage of ascetic fluid into the pleural space through diaphragm defects. The common cause of the hepatopulmonary syndrome and portopulmonary hypertension is portal hypertension and portosystemic shunting, indicating that vasoactive and angiogenetic factors originating from the liver normally control the pulmonary circulation. Portopulmonary hypertension is like pulmonary arterial hypertension, which develops against the background of portal hypertension as a result of chronic liver disease or without other causes of increased pressure in the pulmonary vessels. The prevalence of portopulmonary hypertension ranges from 2% to 8.5% among patients with portal hypertension and is associated with a poor prognosis. Hepatopulmonary syndrome is characterized by intrapulmonary dilatation of microvessels, which causes intrapulmonary shunting and leads to impaired gas exchange in liver diseases, and is associated with a decrease in the quality and duration of life in patients with cirrhosis. Nitric oxide overproduction and angiogenesis seem to be the hallmarks of a complicated pathogenetic mechanism, leading to intrapulmonary shunting and ventilation-perfusion mismatch. A classification of hepatopulmonary syndrome according to the severity of hypoxemia has been suggested. Hepatopulmonary syndrome includes a triad: hepatic dysfunction and / or portal hypertension, dilatation of intrapulmonary vessels, and increased alveolar-arterial oxygen gradient. The prevalence of hepatopulmonary syndrome varies depending on the study groups from 5% to 30%. The most common symptom of the complication is shortness of breath, but in most cases, hepatopulmonary syndrome is asymptomatic. A decrease in oxygen saturation less than 96% corresponds to a decrease in PaO2&lt;70 mm Hg and testifies to the possible development of hepatopulmonary syndrome. In the case of a positive screening, the patient should undergo arterial blood gas analysis, which helps to determine PaO2 and alveolar to arterial oxygen gradient. Conclusion. Contrast-enhanced echocardiography with agitated saline is the gold standard in the diagnosis of intrapulmonary dilatation. The only effective treatment for hepatopulmonary syndrome is liver transplantation. Complete recovery of hepatopulmonary syndrome after liver transplantation is observed within a year in most patients with cirrhosis and hepatopulmonary syndrome

Relationship between endothelin and nitric oxide pathways in the onset and maintenance of hypertension in children and adolescents

The mechanisms that regulate blood pressure are numerous and complex; one mechanism that plays an important role in this scenario is represented by the balance between the vasoconstrictor effect of endothelin-1 and the vasodilator effect of nitric oxide. While there is agreement on the fact that increased endothelin-1 activity and decreased nitric oxide bioavailability are present in hypertensive adults, the situation is less clear in children and adolescents. Not all studies agree on the finding of an increase in plasma endothelin-1 levels in hypertensive children and adolescents; in addition, the picture is often confused by the concomitant presence of obesity, a condition that stimulates the production of endothelin-1. Furthermore, there is recent evidence that, in younger obese and hypertensive subjects, there is an overproduction of nitric oxide, rather than a reduction. This condition may change over time, causing endothelial dysfunction due to a reduced availability of nitric oxide in hypertensive adolescents. The purpose of this review is to address the main biochemical and pathophysiological aspects of endothelin and nitric oxide involvement in hypertension and to summarize the available scientific evidence on their role in the onset and maintenance of high blood pressure in children and adolescents.

The Effect of Acute Exercise on the Interaction between Doxorubicin Administration and Nitric Oxide Metabolism in Heart and Liver Cells

A portion of the reactive species produced as a result of doxorubicin (DOX)-induced oxidative stress is associated with the overproduction of nitric oxide (NO). The subsequent accumulation of DOX in cardiac and liver tissue make these areas particularly susceptible to DOX-induced damage. The purpose of this study was to examine the interaction between DOX administration and exercise on NO metabolism in heart and liver cells. Thirty male Sprague-Dawley rats received an intraperitoneal injection of DOX (4.5 mg/kg) and were randomly separated into 5 experimental groups (n=6). Groups A and E did not perform any exercise and were euthanized at 24 and 48 hours, respectively. Groups B and C underwent a single bout of swimming exercise for 60 min and were sacrificed at 25 and 48 hrs, respectively. Group D exercised at 24 and 48 hrs and were sacrificed at 49 hrs. No changes (P>0.05) in NO concentrations were observed in liver tissue following exercise as compared the non-exercised groups. In contrast, a consistent pattern was observed in cardiac tissues that showed a decrease in NO concentrations in the exercised (B, C, and D) vs. the non-exercised groups (groups A and E). For example, the NO concentrations in the left atrium of group A (non-exercised) was 20±1.1 μmol/mg and decreased (P<0.05) to 15.6±0.6 μmol/mg in group D (exercised). These results suggest that exercise may play a role in lowering DOX induced elevations in NO levels in cardiac tissue, indicating that exercise may assist in mitigating the cardiotoxic effects of chemotherapy.

Nitric Oxide Alters the Pattern of Auxin Maxima and PIN-FORMED1 During Shoot Development.

Hormone patterns tailor cell fate decisions during plant organ formation. Among them, auxins and cytokinins are critical phytohormones during early development. Nitric oxide (NO) modulates root architecture by the control of auxin spatial patterns. However, NO involvement during the coordination of shoot organogenesis remains unclear. Here, we explore the effect of NO during shoot development by using a phenotypic, cellular, and genetic analysis in Arabidopsis thaliana and get new insights into the characterization of NO-mediated leaf-related phenotypes. NO homeostasis mutants are impaired in several shoot architectural parameters, including phyllotactic patterns, inflorescence stem elongation, silique production, leaf number, and margin. Auxin distribution is a key feature for tissue differentiation and need to be controlled at different levels (i.e., synthesis, transport, and degradation mechanisms). The phenotypes resulting from the introduction of the cue1 mutation in the axr1 auxin resistant and pin1 backgrounds exacerbate the relationship between NO and auxins. Using the auxin reporter DR5:GUS, we observed an increase in auxin maxima under NO-deficient mutant backgrounds and NO scavenging, pointing to NO-ASSOCIATED 1 (NOA1) as the main player related to NO production in this process. Furthermore, polar auxin transport is mainly regulated by PIN-FORMED 1 (PIN1), which controls the flow along leaf margin and venations. Analysis of PIN1 protein levels shows that NO controls its accumulation during leaf development, impacting the auxin mediated mechanism of leaf building. With these findings, we also provide evidence for the NO opposite effects to determine root and shoot architecture, in terms of PIN1 accumulation under NO overproduction.

Chromatographic Analysis and In Vitro Cytotoxic Properties of Different Root Extracts of Hermannia geniculata Eckl. &amp; Zeyh on Vero, HepG2 And RAW 264.7 Macrophage

Hermannia geniculata is widely used in the management of several illnesses in South African traditional medicine. Chromatographic analysis, in vitro cytotoxicity, and biological activities of secondary metabolites present in Hermannia geniculata root extracts were investigated. Vero monkey kidney cells, human hepatocellular carcinoma (HepG2) cells, and RAW 264.7 macrophage cell lines were used to determine the cytotoxicity of the extracts using MTT assay. The capabilities of the plant extracts to inhibit 5-lipoxygenase enzyme activities, the overproduction of nitric oxide (NO) following lipopolysaccharide (LPS)-activated RAW 264.7 macrophages by the ethanol extract was evaluated. Results showed selective toxicity of the extracts with LC50 values of Vero cells ranging from (0.40-0.57 mg/mL) while the LC50 value of HepG2 cells varies between (0.016-0.136 mg/mL). The selectivity indexes (SI) were recorded (31.87, 18.87, 33.33, and 13.52) for ethanol, hydro-ethanol, decoction, and aqueous extracts respectively. The ethanol extract inhibited NO production in a concentration-dependent manner showing a decrease of 82% at a concentration of 0.1 mg/mL. Its LC50 value (3.64 mg/mL) is lower and significantly different (p&lt;0.05) compared to quercetin (standard) with an LC50 (8.28 mg/mL). Similarly, the ethanol extract is a potent inhibitor of 5-lipoxygenase enzyme with the lowest IC50 value of 0.14 mg/mL which is significantly different (p&lt;0.05) from other extracts and indomethacin (standard). The GC-MS chromatograms of the ethanol extract revealed five principal compounds that have been reported to have antioxidant, anti-inflammatory, and antifungal properties. This result indicated that Hermannia geniculata root extracts is not toxic to Vero and RAW 264.7 macrophage cell lines and toxic to HepG2 cell lines used in this experiment, it may also possesses antiiflammatory and antiploriferative activities which could be exploited in the development of new, safer, and efficacious drugs. &#x0D; Keywords: Cytotoxicity; Nitric oxide; 5-lipoxygenase; Hermannia geniculata, Antioxidant GCMS.

Hepatic Oxidative Stress, Apoptosis, and Inflammation in Broiler Chickens With Wooden Breast Myopathy.

Wooden breast (WB) syndrome has emerged as a global myopathy in modern commercial broiler chickens, mainly affecting the pectoralis major muscle. Recent evidence suggests that WB myopathy is a systemic disease, which might be accompanied by other physiological disparities and metabolic changes. This study was conducted to systemically investigate the potential physiological changes in liver tissues as well as the possible mechanisms involved to enhance the understanding of the etiology. A total of 93 market-age Arbor Acres male broiler chickens were sampled and categorized into control (CON) and WB groups based on the evaluation of myopathic lesions. Liver samples were collected (n = 10 in each group) for histopathological evaluation and biochemical analyses. Results indicated that WB birds exhibited significantly higher plasma aspartate amino transferase, alkaline phosphatase, and gamma glutamyl transpeptidase activities. Histopathological changes in hydropic/fatty degeneration, inflammatory cell infiltration, intrahepatic hemorrhages, elevated myeloperoxidase activity, and overproduction of nitric oxide were observed in WB liver compared with CON, suggesting the occurrence of liver injury in birds affected by WB myopathy. The WB group showed increased levels of reactive oxygen species, oxidative products, as well as enhanced antioxidant capacities in the liver. These changes were associated with impaired mitochondria morphology and mitochondrial dysfunction. WB myopathy also induced mitochondria-mediated hepatic apoptosis by upregulating levels of caspases 3 and 9, altering the expressions of apoptotic B-cell lymphoma-2 family regulators, as well as increasing the release of cytochrome c. The activation of nuclear factor kappa-light-chain-enhancer of activated B cell signaling enhanced the mRNA expression of downstream inflammatory mediators, contributing to the production of inflammatory cytokines in WB liver. Combined, these findings suggest that hepatic disorders may be conjoined with WB myopathy in broiler chickens and indicating systemic physiological disparities, and other metabolic changes accompanying this myopathy need further assessment.

Insights into the effects of inducible and neuronal nitric oxide synthase isoenzymes in experimental intestinal heterophyiasis

Background: Heterophyiasis is one of the food-borne trematode infections, caused by the intestinal flukeHeterophyes heterophyes. The exact role of nitric oxide (NO) in the immune response against the majorityof parasites remains controversial. It proved protective against a wide range of protozoan and helminthicparasites. Nevertheless, its role in intestinal heterophyiasis is yet to be explored.Objectives: The study aims to explore the possible roles of inducible nitric oxide synthase (iNOS) andneuronal nitric oxide synthase (nNOS) in experimental intestinal heterophyiasis.Material and Methods: The experimental study design included infection of male puppies with H.heterophyes encysted metacercariae (EMC), followed by treatment with aminoguanidine (AG) and7-nitroindazole (7-NI) drugs, as selective inhibitors of iNOS and nNOS, respectively. Controls includednon-infected and infected untreated puppies. Intestinal tissue sections from all puppies were stained forhistopathological and immunohistochemical (IHC) assessments.Results: Different intensities of iNOS and nNOS isoenzymes were observed in intestinal sections. The studyshowed the highest concentration of iNOS isoenzyme in the infected-7-NI treated group. The control noninfectedpuppies exhibited the highest levels of nNOS expression, with statistical significance (P<0.05).The study also showed that AG significantly reduced the degree of inflammatory cellular infiltrations.Additionally, the over-production of NO worsened the degree of intestinal apoptotic changes.Conclusion: Results obtained in the study suggested that inhibition of iNOS, to some extent, improvedintestinal architecture, while inhibition of nNOS failed to eliminate experimental intestinal heterophyiasis.

Identification of 4'-Demethyltangeretin as a Major Urinary Metabolite of Tangeretin in Mice and Its Anti-inflammatory Activities.

The present study showed that oral administration of tangeretin (TAN) in mice resulted in the production of 4'-demethyltangeretin (4DT) as a major urinary metabolite. The anti-inflammatory efficacy of TAN and 4DT was determined in RAW 264.7 macrophages stimulated by lipopolysaccharides (LPS). 4DT produced considerably stronger inhibition on the overproduction of prostaglandin E2 and nitric oxide than TAN did at the same concentrations. Western blot and quantitative polymerase chain reaction analyses indicated that 4DT exerted more potent suppressive activity on the over-expression of interleukin-1β, inducible nitric oxide synthase, and cyclooxygenase-2 than TAN. Treatments with TAN and 4DT diminished LPS-stimulated nuclear factor κB (NFκB) translocation via suppressing the degradation of inhibitor κB (IκBα). Furthermore, both compounds attenuated mitogen-activated protein kinases (MAPKs) and Akt signaling upregulated by LPS. Overall, our findings showed that TAN and 4DT inhibited the LPS-stimulated inflammatory response in macrophages by suppressing Akt/MAPKs/NFκB proinflammatory pathways, while 4DT showed more potent activity than TAN, its parent compound.