• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.113-118
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• 2009

We investigated the structure of bacterial communities present in livestock manure-based composting processes and evaluated the bacterial succession during the composting processes. Compost samples were derived separately from swine manure, dairy manure and sewage sludge. The structure of the bacterial community was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) using universal eubacterial primers. The genus Bacillus and related genera were mainly detected following the thermophilic composting phase of swine and dairy manure composts, and the members of the phylum Bacteroidetes were mainly detected in the cattle manure waste-based and sewage sludge compost. We recovered and sequenced limited number of the bands; however, the PCR-DGGE analysis showed that predominant diversities during the composting processes were markedly changed. Although PCR-DGGE analysis revealed the presence of different phyla in the early stages of composting, the members of the phylum Firmicutes and Bacteroidetes were observed to be one of the predominant phyla after the thermophilic phase.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.12
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• pp.1666-1673
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• 2011

The presence of sialoglycoproteins (SGPs) in the membranes from goat (Capra aegagrus hircus), buffalo (Bubalus bubalis bubalis) and pig (Sus scrofa domestica) erythrocytes was investigated by partial purification with a chloroform-methanol extraction method followed by Sodium dodecyl sulphate - Polyacrylamide gel electrophoresis in comparison to human (Homo sapiens) erythrocytes. The results show that mammalian erythrocytes possess clear differences in the SGPs numbers and molecular weights although all animals studied in this experiment are from the same class i.e. mammalia. The SGPs number in human, goat, buffalo and pig are four (PAS-1 to PAS-4), ten (PAS-GI to PAS-GX), seven (PAS-BI to PAS-BVII) and four (PAS-PI to PAS-IV) respectively as indicated by staining the polyacrylamide gel with sialoglycoprotein-specific Periodic acid-Schiff's (PAS) stain. The new SGPs could be observed only after the partial purification of membrane fractions named as PAS-HI with molecular weight (Mr) 190 kDa and PAS-HII 150 kDa in human, PAS-BIA in buffalo and PAS-PIA and PAS-PIVA in pig. The gels were also stained with Coomassie brilliant blue (CBB) and Silver stain to check the contamination of other membrane proteins in the purified fractions. The quantitative distribution of SGPs was also determined by densitometry. Present study indicates that there are some basic differences in mammalian erythrocyte membrane SGPs, especially with respect to their number and molecular weights indicating major structural variations.

• Korean Journal of Acupuncture
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• v.37 no.2
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• pp.88-96
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• 2020

Objectives : The purpose of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Euphorbia lathyris L. extracted by supercritical CO₂. Methods : Human dermal fibroblast cells dosed with the extract from Euphorbia lathyris L. were harvested and the intracellular proteome was analyzed to examine the expression of proteins related collagen synthesis pathway, metalloproteinases (MMPs), extracellular matrix (ECM)-cell interaction, cytokines, and antioxidant enzymes by 2-dimensional gel electrophoresis. Results : Fatty acid analysis of the extract from Euphorbia lathyris L. showed oleic acid was 84% and linoleic acid was 4.1%. Antioxidative effect was about 53% by beta carotene bleaching assay. In 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis, fifteen protein changes in five mechanisms which were collagen synthesis pathway, MMPs, ECM-cell interaction, cytokines, and antioxidant enzymes were analyzed. Conclusions : This study suggests the supercritical extraction from the seed of Euphorbia lathyris L. could be used as anti-oxidant substances for pharmacopuncture.

• Journal of Radiation Protection and Research
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• v.31 no.2
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• pp.65-68
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• 2006

In this study, the extent of plasmid DNA damage was observed according to concentration of BSH(Boron Sulfhydryl Hydride) and irradiation doses of neutron and ${\gamma}$-ray. The plasmid used was both pBR 322 (2870 bp) and ${\Phi}X174$ RF(5386 bp) DNA. Plasmid DNA damage by irradiation in the presence of BSH was analyzed by agarose gel electrophoresis. In the neutron experiment, DNA damage of both plasmid DNAs was increased according to increasing the concentration of BSH and neutron doses. But in the ${\gamma}$-ray experiment, there appeared no dose dependency as compared to the neutron experiment. The extent of the plasmid DNA damage in the presence of BSH was somewhat different according to irradiation by neutron or ${\gamma}$-ray.

• Tuberculosis and Respiratory Diseases
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• v.77 no.4
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• pp.172-177
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• 2014

Background: Pseudomonas aeruginosa infection is particularly associated with progressive and ultimately chronic recurrent respiratory infections in chronic obstructive pulmonary disease, bronchiectasis, chronic destroyed lung disease, and cystic fibrosis. Its treatment is also very complex because of drug resistance and recurrence. Methods: Forty eight cultures from 18 patients with recurrent P. aeruginosa pneumonia from 1998 to 2002 were included in this study. Two or more pairs of sputum cultures were performed during 2 or more different periods of recurrences. The comparison of strains was made according to the phenotypic patterns of antibiotic resistance and chromosomal fingerprinting by pulsed field gel electrophoresis (PFGE) using the genomic DNA of P. aeruginosa from the sputum culture. Results: Phenotypic patterns of antibiotic resistance of P. aeruginosa were not correlated with their prior antibiotic exposition. Fifteen of 18 patients (83.3%) had recurrent P. aeruginosa pneumonia caused by the strains with same PFGE pattern. Conclusion: These data suggest that the most of the recurrent P. aeruginosa infections in chronic lung disease occurred due to the relapse of prior infections. Further investigations should be performed for assessing the molecular mechanisms of the persistent colonization and for determining how to eradicate clonal persistence of P. aeruginosa.

• Environmental Mutagens and Carcinogens
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• v.21 no.1
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• pp.34-43
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• 2001

Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

• Environmental Mutagens and Carcinogens
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• v.24 no.2
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• pp.79-84
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• 2004

Three synthetic chemicals, benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol were selected for genotoxicity testing, based on production quantity and available genotoxic data. In our previous report, benzoyl chloride induced chromosomal aberrations in Chinese hamster lung (CHL) fibroblast in vitro with and without metabolic activation, while 2-propyn-l-ol and 2-phenoxy ethanol induced only with metabolic activation. To compare the genotoxicity of chromosome aberration assay, the single cell gel electrophoresis (comet) assay subjected using CHL cells. As a result, statistically significant differences of tail moment values of benzoyl chloride, 2-propyn-1-ol, and 2-phenoxy ethanol were observed compared with control values on almost all concentrations with S9 or without S9 metabolic activation system. This results suggest that genotoxic results of the comet assay and the chromosome aberration assay show correlationship of genotoxicity in the CHL fibroblast. In summary, the positive result of chromosome aberration of benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol was also induced DNA damages in comet assay with same cell line. Consequently, comet assay will be useful and more accurate tool to detect and to confirm the genotoxicity especially DNA damages in CHL fibroblast.

• Toxicological Research
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• v.23 no.1
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• pp.25-31
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• 2007

To evaluate the protective effect of Houttuynia cordata on hydrogen peroxide-induced oxidative DNA damage in HepG2 cell line, we used an alkaline single-cell gel electrophoresis (SCGE; comet assay). The DNA damage was analyzed by tail moment (TM) and tail length (TL), which used markers of DNA strand breaks in SCGE. The $100{\mu}g/ml$ of methanolic extract of Houttuynia cordata root showed significant protective effects (p < 0.01) against hydrogen peroxide-induced DNA damage in HepG2 cells and increased cell viability against hydrogen peroxide. The results of this study indicate that Houttuynia cordata root methanol extract acts as a potential antioxidant, and exhibits potential anticancer properties, which may provide a clue to find applications in new pharmaceuticals for oxidative stability.

• Biomolecules & Therapeutics
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• v.13 no.3
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• pp.150-155
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• 2005

The ethanolic extract of chestnut (Castanea crenata S. et Z., Fagaceae) inner shell (CISE) and one of its components, ellagic acid (EA), were evaluated for their protective effects against 1, 1-diphenyl-2-picryl hydrazine (DPPH) free radical generation and hydrogen peroxide-induced oxidative DNA damage in a mammalian cell line. CISE and EA were shown to possess the free radical scavenging effect against DPPH radical generation, significantly. They were also found to strongly inhibit hydrogen peroxide-induced DNA damage from Chinese hamster lung (CHL) cell, assessed by single cell gel electrophoresis assay and 8-hydroxy -2'-deoxy guanosine (8-OH-2'dG) assay. Furthermore, topical application of CISE [$12.5\%$(w/w) cream] and ellagic acid [$1.0\%$(w/w) cream] for 14 days potently inhibited malondialdehyde (MDA) formation of mouse dorsal skin (a marker of lipid peroxidation) induced by ultraviolet B exposure. Therefore, CISE and its component, ellagic acid, may be the useful natural antioxidants by scavenging free radicals, inhibition of lipid peroxidation and protecting oxidative DNA damage when topically applied.

• Journal of Soil and Groundwater Environment
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• v.19 no.5
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• pp.9-17
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• 2014

This paper reported the use of real-time polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and the culture-based method in the intrinsic bioremediation study at a petroleum contaminated site. The study showed that phenol hydroxylase gene was detected in groundwater contaminated with benzene, toluene, ethylbenzene, xylene isomers (BTEX) and methyl tert-butyl ether (MTBE). This indicated that intrinsic bioremediation occurred at the site. DGGE analyses revealed that the petroleum-hydrocarbon plume caused the variation in microbial communities. MTBE degraders including Pseudomonas sp. NKNU01, Bacillus sp. NKNU01, Klebsiella sp. NKNU01, Enterobacter sp. NKNU01, and Enterobacter sp. NKNU02 were isolated from the contaminated groundwater using the cultured-based method. Among these five strains, Enterobacter sp. NKNU02 is the most effective stain at degrading MTBE without the addition of pentane. The MTBE biodegradation experiment indicated that the isolated bacteria were affected by propane. Biodegradation of MTBE was decreased but not totally inhibited in the mixtures of BTEX. Enterobacter sp. NKNU02 degraded about 60% of MTBE in the bioreactor study. Tert-butyl alcohol (TBA), acetic acid, 2-propanol, and propenoic acid were detected using gas chromatography/mass spectrometry during MTBE degraded by the rest cells of Enterobacter sp. NKNU02. The effectiveness of bioremediation of MTBE was assessed for potential field-scale application.