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 (32274 mg 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.

Potential toxicological interactions of 4-(N-methyl-Nnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and/or dibuthyl phthalate (DBP) on ozone were investigated after 32-and 52-wk exposures using hprt mutation assay. Male and female B6C3F1 mice exposed to ozone (0.5 ppm), NNK (1.0 mg/kg), DBP (5,000 ppm), and two or three combinations of these toxicants 6 h per day for 32-and 52-wk showed increases in the frequencies of TG r lymphocytes compared to the control groups. Additive interactions were noted from two combination groups compared to the ozone alone in both sexes of 32-and 52-wk studies. The most common specific mutation type in the hprt genes of test materials-treated male and female mice was transversion with very few transition. The results indicate that such dominant transversion may be responsible for toxicity and combined exposure to ozone, NNK, and DBP induces additive genotoxicities compared to ozone alone.

Objective: The purpose of the current work was to characterize 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 determined by trypan blue exclusion 24 h after treatment, and 6-thioguanineresistant mutants at the xanthine-guanine phosphoribosyltransferase (gpt) gene locus were assessed with doses of which relative survival is 30% or more. Nested PCR-based deletion analysis was also performed.Results: AS52 cells exhibited a dose-dependent increase in cytotoxicity and mutant fraction upon treatment of 2,6-DMA and its metabolites, but showing considerable variation in potency with aminophenol metabolites having the highest potency and parent compound at least; at the highest 2,6-dimethyaminophenol 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 Conclusion: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.. Total deletion of the gpt gene sequences was found in 1 (4%) of spontaneous and 2 (6%) of the 6-thioguanine mutants generated by N-hydroxy-2,6-DMA.

Purpose: To examine the role of endogenous nitric oxide (NO • ) and influence of p53 status during apoptosis induced by a selective iNOS inhibitor, N- [(3-aminomethyl)

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A Satsuma mandarin (Citrus unshiu Marc. cv. Miyagawa) spontaneous mutant, Shinheungri, produces fruit with an abnormal red-coloured peel during ripening. The peel and pulp extracts of Shinheungri fruit were evaluated for polyphenolic contents and antioxidant activities by using various in vitro assays. Compared with those of wild type (WT), Shinheungri exhibited slightly higher total flavonoid content and antioxidant activities. The results of UPLC analyses indicate that the peel and pulp extracts of Shinheungri fruit were rich in mainly hesperidin, and they had different flavonoid composition. The present data on Shinheungri revealed a potential source of powerful flavonoids for further detailed phytochemical investigation.

Dogs have long shared close relationships with many humans. Due to the large number of dogs in human populations, they are often involved in crimes. Occasionally, canine biological evidence such as saliva, bloodstains and hairs can be found at crime scenes. Accordingly, canine DNA can be used as forensic evidence. The use of short tandem repeat (STR) loci from biological evidence is valuable for forensic investigations. In Korea, canine STR profiling-related crimes are being successfully analyzed, leading to diverse crimes such as animal cruelty, dog-attacks, murder, robbery, and missing and abandoned dogs being solved. However, the probability of random DNA profile matches cannot be analyzed because of a lack of canine STR data. Therefore, in this study, 10 STR loci were analyzed in 600 dogs in Korea (344 dogs belonging to 30 different purebreds and 256 crossbred dogs) to estimate canine forensic genetic parameters. Among purebred dogs, a separate statistical analysis was conducted for five major subgroups, 97 Maltese, 47 Poodles, 31 Shih Tzus, 32 Yorkshire Terriers, and 25 Pomeranians. Allele frequencies, expected (Hexp) and observed heterozygosity (Hobs), fixation index (F), probability of identity (P(ID)), probability of sibling identity (P(ID)sib) and probability of exclusion (PE) were then calculated. The Hexp values ranged from 0.901 (PEZ12) to 0.634 (FHC2079), while the P(ID)sib values were between 0.481 (FHC2079) and 0.304 (PEZ12) and the P(ID)sib was about 3.35 × 10−5 for the combination of all 10 loci. The results presented herein will strengthen the value of canine DNA to solving dog-related crimes.

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.