Modulation of A375 human melanoma cell proliferation and apoptosis by nitric oxide

The present study aimed to assess the effect of NO• on melanoma A375 cell growth and apoptotic cell death. Trypan blue exclusion assay was employed to detect the cytotoxicity induced by controlled steady-state concentrations (given in µM • min) of NO•. The characteristics of the cellular cell cycle and apoptosis in NO•-treated A375 cells were also analyzed by Annexin V/PI and DNA fragmentation assays. Western blotting was applied to detect the expression of apoptosis-related proteins (p53, Bax, Fas, DR5, caspase-3 and -9, and PARP). When exposed to preformed 100% NO• for 8 h reactor system, a cumulative dose of 3360 μM • min reduced the viability by 22% 24 h after treatment and promoted apoptosis, 2.9- and 12.2-folds 24 and 48 h after treatment higher than the argon control, respectively. Cell cycle analysis 48 h after treatment revealed S-phase arrest in cells treated with 3360 μM • min NO•. It was also observed that the expression of p53, DR5, caspase 9 and PARP increased significantly upon NO• treatment. In addition, the present study assessed the inhibitory effects of endogenous NO• on the proliferation of human melanoma cells by employing specific (AMG, 1400W and/or SMTC) and nonspecific (NMA) NO• synthase (NOS) inhibitors resulting in melanoma cell growth inhibition; the highest cytotoxic effect was seen when inducible NOS inhibition by 1 mM 1400W treatment. Collectively, the present data suggest that NO• is involved in a key mechanism limiting melanoma proliferation and apoptosis, which may play in improving the efficacy of melanoma treatment

Concert recording 2019-10-29

[Track 1]. From Deux melodies hebraiques. Kaddish / Maurice Ravel -- [Track 2]. From Morike Liederbuch. Der Genesene an die Hoffnung [Track 3]. Denk\u27es, o Seele! [Track 4]. Wo find\u27 ich Trost [Track 5]. Gebet / Hugo Wolf -- [Track 6]. Canata. Prelude Rondo (Peter go ring dem bells) Recitative (Sometimes I feel like a motherless child) Air (Let us break bread together) Toccata (Ride on King Jesus) / John Carter -- [Track 7]. Korean spiritual songs. Yeo-hun wanun nae mokzasini (Jehovah is my Shepherd) Psalm 23 / Unyoung Na -- [Track 8]. Narul badu opsoseo (Lord please take me) / Deok-Shin Choi -- [Track 9]. From Chants de Terre et de Ciel. I. Bail avec Mi [Track 10]. IV. Resurrection / Olivier Messiaen

Phytochemical profile, antioxidant, antimicrobial and antipancreatic lipase activities of fermented Camellia japonica L leaf extracts

Purpose: To investigate the probable antioxidant, antimicrobial and  antipancreatic lipase effects of fermented Camellia japonica leaf extracts.Methods: Camellia japonica leaves fermented with Nuruk were extracted using methanol and ethanol. Total phenolic, flavonoid, carotenoid and L-ascorbic acid contents were determined by UV-visible spectrophotometry. The antioxidant activities of these extracts were determined by free radical scavenging, ferrous ion chelating and reducing power assays. Their  antimicrobial properties against Gram-positive Staphylococcus epidermidis and Bacillus subtilis, and Gram-negative Klebsiella pneumonia and Escherichia coli bacteria were evaluated by disc diffusion method. Inhibition of pancreatic lipase was measured based on the hydrolytic reaction of p-nitrophenyl butyrate with pancreatic lipase.Results: The ethanol extracts of fermented Camellia japonica leaves exhibited higher phenolic (32274mg GAE/100 g) and flavonoid (20519 mg RE/100 g) contents with higher superoxide (IC50 = 0.23  mg/mL), hydrogen peroxide (IC50 = 0.28 mg/mL) radical scavenging and ferrous ion chelating (IC50 = 0.21 mg/mL) activities than those of methanol. These ethanol extracts also showed higher antimicrobial activities against all bacterial strains tested with higher inhibitory effects on pancreatic lipase than methanol extracts.Conclusion: The results highlight the possible use of fermented Camellia japonica leaf extracts as a source of antioxidant, antibacterial and antiobesity agents. Ethanol is recommended as solvent for extracting antioxidants, antibacterial and antiobesity agents from this plant.Keywords: Antioxidant activity, Antimicrobial activity, Fermented Camellia japonica extracts, Pancreatic lipase inhibitio

GENOTOXICITY OF N-HYDROXY AND AMINOPHENOL METABOLITES OF 2,6- AND 3,5-DIMETHYLANILINE AT THE HYPOXANTHINEGUANINE PHOSPHORIBOSYLTRANSFERASE LOCUS IN TK6 CELLS

Objective: The objective of this study as to characterize the genotoxicity of reactive metabolites of 2,6-dimethylaniline (2,6-DMA) and 3,5-DMA in the hypoxanthineguanine phosphoribosyltransferase (HPRT) gene of human lymphoblastoid TK6 cells.Methods: Cultures were exposed to N-hydroxylamine and aminophenol metabolites of 2,6- and 3,5-DMA for 1 h in serum-free medium. Cell survival 24 h after exposure was determined by trypan blue exclusion. Cells were then subcultured for 7–10 days to allow to phenotypic expression of HPRT mutants. After the expression period, cells were plated in the presence of 2 μg/ml 6-thioguanine for the selection of HPRT mutants. Plating efficiency was determined and mutant fraction calculated. Electron paramagnetic resonance (EPR) was also used to determine whether 3,5- dimethylaminophenol (DMAP) produced reactive oxygen species (ROS).Results: All of the metabolites tested were cytotoxic to these cells but exhibited a considerable variation in potency. The aminophenol metabolites of 2,6- and 3,5-DMA were considerably more toxic than the corresponding N-hydroxylamines. Furthermore, each metabolite of 3,5-DMA was more toxic than its 2,6-DMA counterpart; N-OH-3,5-DMA and 3,5-DMAP were clearly mutagenic at a level of 50 μM. EPR studies showed intracellular oxidative stress induced under 3,5-DMAP treatment.Conclusions: Our findings suggest that genotoxic responses of 2,6- and 3,5-DMA are mediated through the generation of ROS by hydroxylamine and/ or aminophenol metabolites.Â

METABOLIC ACTIVATION OF 2,6-DIMETHYLANILINE: MUTATIONAL SPECIFICITY IN THE GPT GENE OF AS52 CELLS

Objective: The purpose of the current work was to characterize the mechanisms of cytotoxicity and mutagenesis of a potential human bladder carcinogen 2,6-dimethylaniline (2,6-DMA).Methods: Chinese hamster ovary (CHO) AS52 cells were exposed to either human S9 activated 2,6-DMA for 6 h or its N-hydroxylamine and aminophenol metabolites for 1 h in serum-free medium. Cell survival was determined by trypan blue exclusion 24 h after treatment, and 6-thioguanine-resistant mutants at the xanthine-guanine phosphoribosyl transferase (gpt) gene locus were assessed with doses, of which relative survival is 30% or more. Nested polymerase chain reaction-based deletion analysis was also performed.Results: AS52 cells exhibited a dose-dependent increase in cytotoxicity and mutant fraction on treatment of 2,6-DMA and its metabolites but show a considerable variation in potency with aminophenol metabolites having the highest potency and parent compound at least; at the highest 2,6-dimethylaminophenol dose (10 μM), the mutant fraction in AS52 cells was 8-fold (13.2×10−5) greater than the spontaneous fraction of 1.62×10−5. Total deletion of the gpt gene sequences was found in 1 (4%) spontaneous and 2 (6%) the 6-thioguanine mutants generated by N-hydroxy-2,6-DMA.Conclusions: These findings indicate the mutagenicity of 2,6-DMA at the gpt gene, which is mediated through hydroxylamine and aminophenol metabolites, and contribute to the elucidation of mechanisms through which 2,6-DMA may exert its effects in vivo

Current-Induced Resonant Motion of a Magnetic Vortex Core: Effect of Nonadiabatic Spin Torque

The current-induced resonant excitation of a magnetic vortex core is investigated by means of analytical and micromagnetic calculations. We find that the radius and the phase shift of the resonant motion are not correctly described by the analytical equations because of the dynamic distortion of a vortex core. In contrast, the initial tilting angle of a vortex core is free from the distortion and determined by the nonadiabaticity of the spin torque. It is insensitive to experimentally uncontrollable current-induced in-plane Oersted field. We propose that a time-resolved imaging of the very initial trajectory of a core is essential to experimentally estimate the nonadiabaticity.Comment: 4 pages, 4 figure