scholarly journals iNOS is not responsible for RyR1 S-nitrosylation in mdx mice with truncated dystrophin.

2020 ◽  
Author(s):  
Ken'ichiro Nogami ◽  
Yusuke Maruyama ◽  
Ahmed Elhussieny ◽  
Fusako Sakai-Takemura ◽  
Jun Tanihata ◽  
...  
Keyword(s):  
Inos Expression ◽  
Mdx Mice ◽  
Transgenic Expression ◽  
Qrt Pcr ◽  
Ryanodine Receptor 1 ◽  
Post Hoc ◽  
Oxide Synthase ◽  
Biotin Switch ◽  
One Way Anova ◽  
Human Dystrophin

Abstract Background: Previous research indicated that nitric oxide synthase (NOS) is the key molecule for S-nitrosylation of ryanodine receptor 1 (RyR1) in DMD model mice ( mdx mice) and that both neuronal NOS (nNOS) and inducible NOS (iNOS) might contribute to the reaction because nNOS is mislocalized in the cytoplasm and iNOS expression is higher in mdx mice. We investigated the effect of iNOS on RyR1 S-nitrosylation in mdx mice and whether transgenic expression of truncated dystrophin reduced iNOS expression in mdx mice or not. Methods: Three- to 4-month-old C57BL/6J, mdx , and transgenic mdx mice expressing exon 45-55-deleted human dystrophin (Tg/ mdx mice) were used. We also generated two double mutant mice, mdx iNOS KO and Tg/ mdx iNOS KO to reveal the iNOS contribution to RyR1 S-nitrosylation. nNOS and iNOS expression levels in skeletal muscle of these mice were assessed by immunohistochemistry (IHC), qRT-PCR, and Western blotting. Total NOS activity was measured by a citrulline assay. A biotin-switch method was used for detection of RyR1 S-nitrosylation. Statistical differences were assessed by one-way ANOVA with Tukey-Kramer post-hoc analysis. Results: mdx and mdx iNOS KO mice showed the same level of RyR1 S-nitrosylation. Total NOS activity was not changed in mdx iNOS KO mice compared with mdx mice. iNOS expression was undetectable in Tg/ mdx mice expressing exon 45-55-deleted human dystrophin, but the level of RyR1 S-nitrosylation was the same in mdx and Tg/ mdx mice. Conclusion: Similar levels of RyR1 S-nitrosylation and total NOS activity in mdx and mdx iNOS KO demonstrated that the proportion of iNOS in total NOS activity was low, even in mdx mice. Exon 45-55-deleted dystrophin reduced the expression level of iNOS, but it did not correct the RyR1 S-nitrosylation. These results indicate that iNOS was not involved in RyR1 S-nitrosylation in mdx and Tg /mdx muscles.

scholarly journals iNOS is not responsible for nitrosylation of RyR1 in mdx mice with truncated dystrophin

2020 ◽  
Author(s):  
Ken'ichiro Nogami ◽  
Yusuke Maruyama ◽  
Ahmed Elhussieny ◽  
Fusako Sakai-Takemura ◽  
Jun Tanihata ◽  
...  
Keyword(s):  
Inos Expression ◽  
Mdx Mice ◽  
Transgenic Expression ◽  
Qrt Pcr ◽  
Ryanodine Receptor 1 ◽  
Post Hoc ◽  
Oxide Synthase ◽  
Biotin Switch ◽  
One Way Anova ◽  
Human Dystrophin

Abstract Background: Previous research indicated that nitric oxide synthase (NOS) is the key molecule for S-nitrosylation of ryanodine receptor 1 (RyR1) in DMD model mice (mdx mice) and that both neuronal NOS (nNOS) and inducible NOS (iNOS) might contribute to the reaction because nNOS is mislocalized in the cytoplasm and iNOS expression is higher in mdx mice. We investigated the effect of iNOS on RyR1 S-nitrosylation in mdx mice and whether transgenic expression of truncated dystrophin reduced iNOS expression in mdx mice or not.Methods: Three- to 4-month-old C57BL/6J, mdx, and transgenic mdx mice expressing exon 45-55-deleted human dystrophin (Tg/mdx mice) were used. We also generated two double mutant mice, mdx iNOS KO and Tg/mdx iNOS KO to reveal the iNOS contribution to RyR1 S-nitrosylation. nNOS and iNOS expression levels in skeletal muscle of these mice were assessed by immunohistochemistry (IHC), qRT-PCR, and Western blotting. Total NOS activity was measured by a citrulline assay. A biotin-switch method was used for detection of RyR1 S-nitrosylation. Statistical differences were assessed by one-way ANOVA with Tukey-Kramer post-hoc analysis.Results: mdx and mdx iNOS KO mice showed the same level of RyR1 S-nitrosylation. Total NOS activity was not changed in mdx iNOS KO mice compared with mdx mice. iNOS expression was undetectable in Tg/mdx mice expressing exon 45-55-deleted human dystrophin, but the level of RyR1 S-nitrosylation was the same in mdx and Tg/mdx mice.Conclusion: Similar levels of RyR1 S-nitrosylation and total NOS activity in mdx and mdx iNOS KO demonstrated that the proportion of iNOS in total NOS activity was low, even in mdx mice. Exon 45-55-deleted dystrophin reduced the expression level of iNOS, but it did not correct the RyR1 S-nitrosylation. These results indicate that iNOS was not involved in RyR1 S-nitrosylation in mdx and Tg/mdx mice muscles.

scholarly journals Anti-Inflammatory Effect of Phytoncide in an Animal Model of Gastrointestinal Inflammation

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 1895
Author(s):  
Azra Memon ◽  
Bae Yong Kim ◽  
Se-eun Kim ◽  
Yuliya Pyao ◽  
Yeong-Geun Lee ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Animal Model ◽  
Gastric Ulcer ◽  
Dextran Sulfate ◽  
Dextran Sulfate Sodium ◽  
Inos Expression ◽  
Gastrointestinal Inflammation ◽  
Anti Inflammatory ◽  
Oxide Synthase ◽  
Inflammatory Effect

Background: Phytoncide is known to have antimicrobial and anti-inflammatory properties. Purpose: This study was carried out to confirm the anti-inflammatory activity of two types of phytoncide extracts from pinecone waste. Methods: We made two types of animal models to evaluate the efficacy, an indomethacin-induced gastroenteritis rat model and a dextran sulfate sodium-induced colitis mouse model. Result: In the gastroenteritis experiment, the expression of induced-nitric oxide synthase (iNOS), a marker for inflammation, decreased in the phytoncide-supplemented groups, and gastric ulcer development was significantly inhibited (p < 0.05). In the colitis experiment, the shortening of the colon length and the iNOS expression were significantly suppressed in the phytoncide-supplemented group (p < 0.05). Conclusions: Through this study, we confirmed that phytoncide can directly inhibit inflammation in digestive organs. Although further research is needed, we conclude that phytoncide has potential anti-inflammatory properties in the digestive tract and can be developed as a functional agent.

scholarly journals Activation of Nuclear Factor κB and Induction of Inducible Nitric Oxide Synthase by Ureaplasma urealyticumin Macrophages

2000 ◽  
Vol 68 (12) ◽  
pp. 7087-7093 ◽  
Author(s):  
Y.-H. Li ◽  
Z.-Q. Yan ◽  
J. Skov Jensen ◽  
K. Tullus ◽  
A. Brauner
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Inducible Nitric Oxide Synthase ◽  
Alveolar Macrophages ◽  
Nuclear Factor Κb ◽  
Inos Expression ◽  
Dependent Manner ◽  
Nuclear Factor ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

ABSTRACT Chronic lung disease (CLD) of prematurity is an inflammatory disease with a multifactorial etiology. The importance ofUreaplasma urealyticum in the development of CLD is debated, and steroids produce some improvement in neonates with this disease. In the present study, the capability of U. urealyticum to stimulate rat alveolar macrophages to produce nitric oxide (NO), express inducible nitric oxide synthase (iNOS), and activate nuclear factor κB (NF-κB) in vitro was characterized. The effect of NO on the growth of U. urealyticum was also investigated. In addition, the impact of dexamethasone and budesonide on these processes was examined. We found that U. urealyticum antigen (≥4 × 107 color-changing units/ml) stimulated alveolar macrophages to produce NO in a dose- and time-dependent manner (P < 0.05). This effect was further enhanced by gamma interferon (100 IU/ml; P < 0.05) but was attenuated by budesonide and dexamethasone (10−4 to 10−6 M) (P < 0.05). The mRNA and protein levels of iNOS were also induced in response to U. urealyticum and inhibited by steroids.U. urealyticum antigen triggered NF-κB activation, a possible mechanism for the induced iNOS expression, which also was inhibited by steroids. NO induced by U. urealyticum caused a sixfold reduction of its own growth after infection for 10 h. Our findings imply that U. urealyticum may be an important factor in the development of CLD. The host defense response againstU. urealyticum infection may also be influenced by NO. The down-regulatory effect of steroids on NF-κB activation, iNOS expression, and NO production might partly explain the beneficial effect of steroids in neonates with CLD.

scholarly journals 16K prolactin induces NF-kappaB activation in pulmonary fibroblasts

2002 ◽  
Vol 175 (3) ◽  
pp. R13-R18 ◽  
Author(s):  
Y Macotela ◽  
C Mendoza ◽  
AM Corbacho ◽  
G Cosio ◽  
JP Eiserich ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nuclear Translocation ◽  
Signal Transduction Pathway ◽  
Binding Activity ◽  
Inos Expression ◽  
Pulmonary Fibroblasts ◽  
Amino Terminal ◽  
Dna Binding Activity ◽  
Nf Kappab ◽  
Oxide Synthase

The amino-terminal 16 kDa fragment of prolactin (16K PRL) promotes the expression of the inducible isoform of nitric oxide synthase (iNOS) accompanied by the production of nitric oxide (NO) by rat pulmonary fibroblasts. The present study was designed to elucidate whether the mechanism by which 16K PRL promotes iNOS expression involves the activation of nuclear factor-kappa B (NF-kappaB), a key transcription factor for iNOS induction. 16K PRL stimulated DNA-binding activity of NF-kappaB in pulmonary fibroblasts as demonstrated by gel shift assays. Likewise, fluorescence immunocytochemistry showed that 16K PRL promotes nuclear translocation of the p65 subunit of NF-kappaB. Finally, treatment with 16K PRL induced the degradation of the NF-kappaB inhibitor kappaB-beta (IkappaB-beta), and such degradation was prevented by blocking IkappaB-beta phosphorylation. Altogether, these results show that 16K PRL activates NF-kappaB nuclear translocation via the phosphorylation and degradation of IkappaB-beta. These findings are consistent with NF-kappaB being part of the signal transduction pathway activated by 16K PRL to induce iNOS expression.

Abstract 316: Effects of Inhibiting of Inducible Nitric Oxide Synthase in the Pathophysiology of Experimental Preeclampsia

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Lorena M Amaral ◽  
Ana Carolina T Palei ◽  
Lucas C Pinheiro ◽  
Jonas T Sertorio ◽  
Danielle A Guimaraes ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Reactive Oxygen Species ◽  
Mean Arterial Pressure ◽  
Arterial Pressure ◽  
Inducible Nitric Oxide Synthase ◽  
Inos Expression ◽  
Oxygen Species ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

The pathophysiology of preeclampsia (PE) is not entirely known. However, increased oxidative stress possibly leading to impaired nitric oxide activity has been implicated in the critical condition. Increased oxidative stress with increased levels of highly reactive species including superoxide may generate peroxynitrite. We examined the role of inducible nitric oxide synthase (iNOS) and oxidative stress in the reduced uterine perfusion pressure (RUPP) preeclampsia experimental model. METHODS: RUPP was induced in wistar rats. Pregnant rats in the RUPP group had their aortic artery clipped at day 14 of gestation. After a midline incision, a silver clip (0.203 mm) was placed around the aorta above the iliac bifurcation; silver clips (0.100 mm) were also placed on branches of both the right and left ovarian arteries that supply the uterus. Sham-operated (pregnant control rats) and RUPP rats were treated with oral vehicle or 1 mg/kg/day 1400W (iNOS inhibitor) for 5 days. Mean arterial pressure (MAP) and plasma levels of thiobarbituric acid-reactive species (TBARS) and total radical-trapping antioxidant potential (TRAP) were measured determined. Aortic iNOS expression (Western blotting) and reactive oxygen species (ROS; assessed by fluorescence microscopy with dihydroethidium-DHE) were measured. We found increased mean arterial pressure in RUPP compared with pregnant control rats (MAP= 128±1 vs. 100±1.8 mmHg, respectively; P<0.05) and 1400W exerted antihypertensive effects (MAP= 114±2 vs.128±1 mmHg in RUPP treated and untreated rats, respectively; P<0.05). Higher reactive oxygen species (ROS) concentrations were found in RUPP compared with pregnant control rats (7.1±0.5 vs. 5.1±0.5 arbitrary units (A.U.), respectively; P<0.05) and 1400W decreased ROS production to 5.8±0.02 A.U. in RUPP treated rats, P<0.05. In addition, 1400W attenuated iNOS expression in RUPP rats (0.29±0.02 vs. 0.55±0.8 A.U. in RUPP treated and untreated rats, respectively; P<0.01) and had no effects on plasma TBARS and TRAP levels. Our results suggest that 1400w exerts antihypertensive effects in the RUPP model and suppresses ROS formation. Supported by FAPESP,Cnpq.

scholarly journals Immune rejection of human dystrophin following intramuscular injections of naked DNA in mdx mice

Gene Therapy ◽  
2000 ◽  
Vol 7 (17) ◽  
pp. 1447-1457 ◽  
Author(s):  
S Braun ◽  
C Thioudellet ◽  
P Rodriguez ◽  
D Ali-Hadji ◽  
F Perraud ◽  
...  
Keyword(s):  
Mdx Mice ◽  
Immune Rejection ◽  
Naked Dna ◽  
Intramuscular Injections ◽  
Human Dystrophin

scholarly journals Depth-Related Curing Potential of Ormocer- and Dimethacrylate-Based Bulk-Fill Composites

Materials ◽  
2021 ◽  
Vol 14 (22) ◽  
pp. 6753
Author(s):  
Ramona S. Oltramare ◽  
Reto Odermatt ◽  
Phoebe Burrer ◽  
Thomas Attin ◽  
Tobias T. Tauböck
Keyword(s):  
Double Bond ◽  
Repeated Measures ◽  
In Vitro Study ◽  
High Viscosity ◽  
Degree Of Conversion ◽  
Double Bond Conversion ◽  
Nanohybrid Composite ◽  
Post Hoc ◽  
One Way Anova

The aim of this in vitro study was to investigate the degree of C=C double bond conversion of high-viscosity dimethacrylate- or ormocer-based bulk-fill composites as a function of measurement depth. Four bulk-fill composites (Tetric EvoCeram Bulk Fill, x-tra fil, SonicFill, and Bulk Ormocer) and the conventional nanohybrid composite Tetric EvoCeram were applied in standardized Class II cavities (n = 6 per group) and photoactivated for 20 s at 1350 mW/cm2. The degree of conversion of the composites was assessed using Fourier-transform infrared spectroscopy at seven measurement depths (0.15, 1, 2, 3, 4, 5, 6 mm). Data were analyzed using repeated measures ANOVA and one-way ANOVA with Bonferroni post-hoc tests (α = 0.05). The investigated bulk-fill composites showed at least 80% of their maximum degree of conversion (80% DCmax) up to a measuring depth of at least 4 mm. Tetric EvoCeram Bulk Fill and Bulk Ormocer achieved more than 80% DCmax up to a measuring depth of 5 mm, x-tra fil up to 6 mm. The conventional nanohybrid composite Tetric EvoCeram achieved more than 80% DCmax up to 3 mm. In contrast to the conventional composite, the investigated ormocer- and dimethacrylate-based bulk-fill composites can be photo-polymerized in thick layers of up to at least 4 mm with regard to their degree of C=C double bond conversion.

scholarly journals Avaliação e proposta de monitoramento do estado de conservação de ambiente recifal costeiro do Estado da Paraíba, Brasil: contribuições para gestão ambiental

2018 ◽  
Vol 5 (10) ◽  
pp. 949-967 ◽  
Author(s):  
Daniel Silva Lula Leite ◽  
George Emmanuel Cavalcanti de Miranda
Keyword(s):  
Gelidiella Acerosa ◽  
Post Hoc ◽  
Canistrocarpus Cervicornis ◽  
One Way Anova

A urbanização nas zonas costeiras torna necessário avaliar o impacto antrópico sobre os ecossistemas associados. O objetivo deste trabalho foi analisar temporalmente a dinâmica de populações de macroalgas com potencial bioindicador, estabelecendo-as como ferramenta para avaliação da saúde ambiental. Foi amostrado o ambiente recifal intertidal da Praia de Formosa, no Município de Cabedelo, Estado da Paraíba, Nordeste do Brasil, durante intervalos trimestrais de agosto/2013 a novembro/2014. A variação temporal da frequência de ocorrência (F') das populações macrofitobênticas foi avaliada pela análise de variância (One-Way Anova) com teste Post-Hoc de Tukey. A influência das variáveis ambientais sobre F’ foi avaliada pela Correlação Linear de Pearson e pela PERMANOVA. A análise da qualidade ambiental e a proposta de biomonitoramento basearam-se no Grupo 1, formado por espécies perenes e/ou raras com maiores valores ao longo do Componente Principal 1 (CP1) da Análise de Componentes Principais (ACP), e Grupo 2, formado por espécies com comportamento errático com menores valores ao longo do CP1 da ACP. Foram encontradas 33 espécies macrofitobênticas. A frequência de ocorrência apresentou variação temporal, porém não influenciada pelo conjunto de variáveis ambientais. Phyllodictyon anastomosans, Gelidiella acerosa e Gelidium corneum compuseram o Grupo 1, com Dictyopteris delicatula, Canistrocarpus cervicornis e Hypnea spinella formando o Grupo 2. O Grupo 2 apresentou correlação positiva com a temperatura, alcançando as máximas frequências nos períodos mais quentes, mas esse fator isolado não é capaz de explicar a distribuição errática das espécies. A ocorrência de espécies bioindicadoras de eutrofização foi considerada natural, fato que, aliado à alta frequência de espécies bioindicadoras de qualidade ambiental, sugerem que o Recife de Formosa não sofre impactos antrópicos significativos. O desaparecimento de espécies perenes ou crescimento populacional significativo de espécie rara (Grupo 1), bem como a fuga do padrão temporal de frequência ou dominância do recife pelas espécies do Grupo 2, são indícios da ocorrência de impactos. Este trabalho fornece à gestão do Recife de Formosa uma eficiente ferramenta para avaliação do impacto urbano sobre a comunidade macrofitobêntica.

scholarly journals The Robusta coffee grounds residues to adsorb the heavy metal Lead (Pb) in the water

2018 ◽  
Author(s):  
Anita Dewi Moelyaningrum ◽  
Khoirul Ngibad ◽  
S.Pd Lilla Puji Lestari ◽  
Dessy Agustina Sari ◽  
Cahya Fajar Budi Hartanto ◽  
...  
Keyword(s):  
High Carbon ◽  
Control Groups ◽  
Robusta Coffee ◽  
Randomized Design ◽  
Coffee Grounds ◽  
Coffee Ground ◽  
Completely Randomized Design ◽  
Post Hoc ◽  
One Way Anova ◽  
Ks Test

This paper has been published in IOP Conf. Series: Journal of Physics: Conf. Ser1i2e3s415161748(9200‘1’8“)”012058 Lead (Pb) were toxic. Lead found in pipes, batteries, paint, dyes ceramic glaze, gasoline, and final waste disposal. The robusta coffee grouds residues had high carbon, nitrogen etc which can adsorb heavy metal.The purpose of this study is to analyze the robusta coffee grounds residues to adsorb the Pb in the water. The method of this research is a True Experimental using completely randomized design (CRD) method. There were control groups (C) and three treatments groups (T1; T2;T3; 5gram/ litre; 8gram/ litre; 10gram/ litre) with six repetitions. The robusta coffee ground residues were contacted for 2 hours.Total samples were24 samples which analyzed each parameter of the Pb with Atomic Adsorption Spectophotometry Analysis. The results showed that the more coffee ground residues that are exposed, increasingly turbid. The KS test showed that data were a normal distribution (sig=0,324). One way ANOVA test; Turkey post Hoc showed that there was sig difference between the control and treatment (F=4,326, Sig= 0,017). There were sig difference between control and treatment 2 and 3 (Sig=0,019; Sig=0,038). Robusta coffee grounds residues can reuse to adsorb the Pb pollution in the water. It can be a solution for treating the lead pollution in the water because of it easy to the application.

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