• The Korean Journal of Mycology
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• v.44 no.3
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• pp.138-144
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• 2016

Twelve unrecorded yeasts, Pseudozyma prolifica HL9-1, Trichosporon coremiiforme NS19-2, Candida cretensis SA4-1, Cryptococcus diffluens TJ4-3, Cryptococcus pinus YB17-2, Candida vartiovaarae DD2-5, Pichia galeiformis DM3-5, Candida pseudolambica JW2-3, Trichosporon xylopini NS5-1, Trichosporon moniliiforme NS5-7, Tetrapisispora iriomotensis NS14-2, and Tetrapisispora nanseiensis SA17-1, were screened among 97 yeasts from soils of Chungcheongnam-do and Daejeon metropolitan city, Korea. These yeasts were oval or ellipsoidal and had a budding system for vegetative reproduction. They grew well in yeast extract-peptone-dextrose (YPD) medium and, in particular, Tetrapisispora iriomotensis NS14-2 and Candida cretensis SA4-1 grew well in 10% NaCl-containing YPD broth. Nine strains, including Trichosporon coremiiforme NS19-2, assimilated xylose and four yeast strains, such as Candida vartiovaarae DD2-5, also assimilated lactose. Physiological functionalities of cell-free extracts and supernatants from two halophilic unrecorded yeasts, Candida cretensis SA4-1 and Tetrapisispora iriomotensis NS14-2, were investigated. Cell-free extracts from Candida cretensis SA4-1 and Tetrapisispora iriomotensis NS14-2 exhibited 71.3% and 68.4% antihypertensive angiotensin I-converting enzyme inhibitory activity.

• The Journal of Korean Medicine
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• v.19 no.2
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• pp.373-390
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• 1998

35 theses conducted in Korea on the topic of senescence or anti-oxidant were classified on the basis of research methods, animals used for the experiments, and research items. Evaluating these research works with respect to the Free Radical Theory, the following conclusions were reached. 1. Of the 17 theses written in the Oriental medicine aspect, three theses used a single herb, nine theses used a complex prescription, four theses concentrated on the usage of the medicinal acupuncture, and one research paper focused on using scientific components. Common objection of these papers were on the verification of the efficacy of herbs. 2. Of the 18 these written in the Western medicine aspect, five theses used a single drug, seven theses conducted a research on the changes due to senescence, and 6 papers were on variety of topics. The main focus of these works were on the mechanism and pathology related to the senescence rather than on the suppression of senescence. 3. Among the theses written in the Oriental medicine perspective, a total of 48 herbs were utilized. 26 of these herbs has a tonification function on the Kidney. Six out of nine complex prescription mentioned above has a function of tonifying the Kidney. 4. With respect to the research subjects used on the experiments, 8 theses have used Senescence-Accelerated mice, 13 theses have used Sprague-Dawley mice, and remaining 7 papers have used human or other animals. 5. These are the categorization of the research items used: the weight (11) and weight changes of the visceral organs (9), the measurement of the content of peroxide-disease (9), the measurement of enzyme vitality (21), the blood and urine test (10), the experiment concerning immune system (3), the influence on the hepatic capability of metabolizing foreign substance (3), the effect on hepatic cell protection (3), the measurement of both the suppression of Free Radical and ability to create Free Radical (2), the measurement of effect of suppresing MDA(malondialdehyde) (4), the effect of eliminating DPPH Radical (2), and experiements about the functions (2). 6. The rate of vitalization of well known anti-oxidants such as Superoxide dismutase (SOD), Protein-bound SH, Nonprotein-bound SH, Glutathione(GSH), Catalase, and etc. were tested in 17 theses. Considering the conclusions mentioned above, the theses related to the senescence published in Korea elected different animals used for experiments, research items and the methods of research, the end result seems to be a lack of objectivity. Thus, I would argue that research methods to overcome such a deficiency need to be developed systematically.

• Journal of Nutrition and Health
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• v.33 no.7
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• pp.725-732
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• 2000

The purpose of this study was to investigate the effects of green tea catechin on renal dysfunction and blood presure change in chronic cadmium poisoned rats. Sprague-Dawley male rats weighing 100$\pm$10g were randomly assigned to one normal group and three cadmium poisoned groups. Cadmium groups were classified to catechin free diet(Cd-0C group) 0.25% catechin diet(Cd-0.25C group) and 0.5% catechin diet(Cd-0.5C group) according to the levels of catechin supplement. Animals were raids for 20weeks. Cadmium were supplied as drinking water of 50ppm Cd2+ Morphological changes shown through a light microscope and an electro-microscope revealed the mitochondria and tubule epithelial cell edema in Cd -0C group but they were alleviated in catechin supplementation. The urinary $\beta$2-microglobulin that measured to observe the glomerular injury were higher in Cd-poisoned groups than in normal group but they was lowered by catechin supplementation. Glomerular filtration ratios(GFR) in Cd-poisoned groups were significantly lower than in normal group but that of catechin supplementation group was similar to normal group. This suggested that catechin protected the kidney from the functional damage. Angiotensin converting enzyme(ACE) activity and blood pressure(BP) in Cd-poisoned groups were significantly higher than in normal group. Heart rate was tended to increase in Cd-poisoned groups. The results indicate that green tea catechin supplementation on chronic cadmium-poisoned rats normalized the renal dysfunction and blood pressure system.

• Microbiology and Biotechnology Letters
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• v.14 no.5
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• pp.427-432
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• 1986

The regulation of three ammonia assimilatory enzymes, GDH (glutamate dehydrogenase), GS (glutamine synthetase) and GOGAT (glutamate synthase), has been examined in C. glutamicum. Three kinds of arginine auxotrophs blocked in each step of arginine biosynthetic pathway from glutamate were selected as arg 5, arg 6, arg 8. Histidine and tryptophan auxotrophs were also selected because histidine and tryptophan repressed GS biosynthesis in E. coli. These strains were cultured on the media containing nitrogen-excess and limited conditions, to compare the specific activities of ${\alpha}$-ketoglutarate dehydrogenase(${\alpha}-KGDH$), GDH, GS, GOGAT from the cell-free extracts. These results showed that enzyme levels of ${\alpha}-KGDH$ and GDH from 3 kinds of arginine auxotrophs, histidine and tryptophan auxotrophs in nitrogen-excess condition and those of GS and GOGAT in nitrogen limited condition were increased compared with opposite condition. The tryptophan and histidine auxotrophs showed higher level of glutamate and glutamine than parental strains and other mutants. it is assumed that the higher levels of ${\alpha-KGDH}$ and GDH from mutants in nitrogen-excess condition promoted the accumulation of glutamate and glutamine in fermentation broth. The inhibition of GS activities by ADP suggested that GS is regulated by energy charge in C. glutamicum. The results with histidine, tryptophan, glycine, alanine, serine and GMP implied that a system of feedback inhibition were effective. The GDH, GS and GOGAT biosynthesis in culture broth was markedly repressed by the nature and kinds of available nitrogen sources such as tryptophan, proline, glycine, alanine, serine and tyrosine.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.4
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• pp.446-450
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• 2000

The experiment was conducted to study the effects of water soluble vitamins and retinoic acid on the differentiation of preadipocyte from omental, subcutaneous, intermuscular and intramuscular adipose tissue of Hanwoo. Differentiation was assessed by the change in enzyme activity, glycerol-3 phosphate dehydrogenase in serum free cell culture system. Preadipocytes treated with biotin ($10{\mu}M$) and pantothenic acid ($100{\mu}M$) were significantly (p<0.05) less differentiated than those from the control in all adipose tissue depots except intramuscular tissue. Although there was no significance, vitamin C was shown to stimulate the adipocyte conversion in omental and subcutaneous, but not in intermuscular and intramuscular adipose tissues. Lower values of GPDH activity in intermuscular preadipocyte were interpreted to be caused by relatively higher amounts of protein. In this experiment vitamin C did not stimulate fat deposition in intramuscular adipose tissue but further experiments are needed on the role of vitamin C in preadipocyte differentiation. When treated with different levels of retinoic acids, differentiation of preadipocytes was significantly (p<0.05) reduced from the level of $0.5{\mu}g/ml$ in omental and intermuscular, from $50{\mu}g/ml$ in subcutaneous, and in intramuscular at $500{\mu}g/ml$, thus showing that intramuscular preadipocytes were least responsive to retinoic acid in differentiation. All-trans retinoic acid appeared to inhibit the differentiation in a dose dependent manner, regardless of adipose tissues type.

• Journal of Environmental Health Sciences
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• v.30 no.3
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• pp.230-238
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• 2004

The adverse health effects of a number of environment pollutions are related to the formation of free radicals. Induction of antioxidant defensive system in the response to an oxidative attack is an essential element of the cell to survive. CYP2E1 is easily induced by organic solvents and induces continuous formation of reactive oxygen species (ROS). ${\gamma}$-Glutamyltranspeptidase (${\gamma}$GT) plays an important role in glutathione metabolism and xenobiotic detoxification. To evaluate the characteristic of oxidative stress which induces GGT expression and to understand human antioxidant defensive response against oxidative stress induced by CYP2E1, we studied regulation of ${\gamma}$GT enzyme expression in response to various oxidative stresses in human HepG2 cells. The ${\gamma}$GT activity was not modified after exposure of acute oxidative stress inducing agents (ferric nitrilotriacetate, cumene hydroperoxide, ADP-Fe, O-tetradecanoylphorbol-13-acetate, tumor necrosis factor-alpha). To induce continuous exposure of cells to ROS, HepG2 cells were transfected by human CYP2E1 gene transiently. The CYP2E1 activity was verified with chlorzoxazone hydroxylation. Transfection of CYP2E1 showed continuous 60% increase in intracellular ROS and 240 % increase in microsomal ROS. CYP2E1 overexpressing cells showed increased ${\gamma}$GT activity (2.5-fold). The observed enhancement of ${\gamma}$GT activity correlated with a significant increase of ${\gamma}$GT mRNA (2.1-fold). Treatment with antioxidant strongly prevented the increase in ${\gamma}$GT activity. The CYP2E1 overexpression did not modify toxicity index and increased glutathione levels. These results show that continuous exposure of cells to ROS produced by CYP2E1 up-regulates ${\gamma}$GT; This may be one of the adaptive antioxidant responses of cells to oxidative insult. Present study also suggests that the induction of ${\gamma}$GT could be used as a marker of oxidative stress induced by exposure to organic solvents.

• Korean Journal of Environmental Agriculture
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• v.24 no.2
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• pp.146-152
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• 2005

The objective of present study was to investigate the effect of onion extracts on mercuryinduced cytotoxicity, lipid peroxidation and antioxidant enzyme activities in primary monolayer cultures of rat hepatocytes. Primary cultures of rat hepatocytes were incubated for 6 hr in the presence of various concentrations (0, 1, 5, 10, 30 or 50 ppm) of $HgCl_2$. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase (GOT) activity, lactate dehydrogenase (LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) value. Lipid peroxidation w as evaluated using thiobarbituric acid reactive substances (TBARS) assay. Effects of onion extract on antioxidant system were determined by measuring catalase, glutathione peroxidase (GSH-Px), glutathione reductase (GSH-Rd) activities as well as DPPH free radical scavenging activity. $HgCl_2$ at the concentration of 10 ppm increased GOT activity and TBARS concentration but decreased %MTT reduction, whereas $HgCl_2$ at the concentration of 30 ppm increased LDH activity, representing that $HgCl_2$ caused cytotoxicity and lipid peroxidation in dose-dependent manner, $HgCl_2$ at the concentration of 30 ppm significantly decreased catalase, GSH-Px and GSH-Rd activities. When primary cultures of rat hepatocytes were incubated with various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract for 6 hr in the presence of 30 ppm of $HgCl_2$, onion extracts at the concentration of 0.05 mg/ml decreased GOT activity, but increased %MTT reduction by 30 ppm of $HgCl_2$. $HgCl_2-induced$ LDH activity and TBARS concentration were decreased by onion extract at the concentration of 0.01 mg/ml. Taken together, onion extract prevented H$HgCl_2-induced$ hepatocyte injury and lipid peroxidation. Onion extracts at the concentration of 0.1 mg/ml almost or completely inhibited $HgCl_2-induced$ catalase and GSB-Px activities. GSH-Rd activity, however, was not affected by onion extract. Free radical scavengjing activity was increased as concentration of onion extract increased. Onion extract at the concentrion of 5 mg/ml possesed mote than 93% scavenging activity comparing to 100% radical scavenging activity by pyrogallol solution as a reference. These results demonstrate that onion extracts suppressed mercury-induced cytoctoxicity and lipid peroxidation by scavenging free radical and increasing catalase and GSH-Px activities.

• Food Science and Preservation
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• v.23 no.6
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• pp.890-898
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• 2016

The purpose of this paper is to investigate potential anti-inflammatory and anti-oxidant effects of Tenebrio molitor. Macrophage cell response by outside stimulation leads expression of pro-inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), $interleukin-1{\beta}$ ($IL-1{\beta}$), and trigger expression of genes which are affected by inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), resulting in formation of inflammatory factors like nitric oxide (NO) and Prostaglandin $E_2$ (PGE2). Cell viability was determined by MTT assay. In order to investigate anti-inflammatory agents, the inhibitory effects on the production of lipopolysaccharide (LPS)-induced NO in RAW 264.7 cells were examined. T. Molitor significantly decreased the production of NO in a dose-dependent manner, and also reduced the expression of iNOS, a COX-2 protein. As a result, the levels of protein such as $PGE_2$, iNOS, COX-2 and MARKs were significantly reduced compared to non-treated group in T. Molitor water extract (TDW) treated group. Also, antioxidant effect of T. Molitor were investigated using DPPH, ABTS+ and superoxide anion radical scavenging activity tests in cell-free system. Antioxidant activity of T. molitor was found low in the DPPH radical scavenging test while high in the ABTS+ and superoxide anion radical scavenging activity tests. These results show that TDW could be an effective anti-pro-inflammatory and anti-oxidant agent.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.4
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• pp.287-292
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• 2009

Human skin is constantly exposed to environmental irritants such as smoke, chemicals and ultraviolet (UV). Free radicals and reactive oxygen species (ROS) caused by these environmental irritants play critical roles in cellular damage. In this study, to investigate the skin cell protective effect of Ophioglossum vulgatum extract, we investigated its effects on intercellular antioxidative activity and UVA-induced MMP expression in human dermal fibroblasts (HDFs). The dried O. vulgatum was extracted in a mixture of ethanol and water (1 : 1) for 24 h at room temperature. The extract was filtered and concentrated in vacuo and lyophilized. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB $20\;mJ/cm^2$. After treatment of O. vulgatum extracts, intracellular ROS levels were measured by luminescence spectrophotometer. Enzyme linked immuno sorbent assay (ELISA), and RT-PCR techniques were used for evaluating the effects of O. vulgatumon on MMP protein and mRNA expression in UVA irradiated HDFs. O. vulgatum extract was found to have ROS scavenging activity with the $IC_{50}$ values of $18.2\;{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system. After treatment of O. vulgatum extracts, the oxidation of CM-DCFDA was inhibited effectively and O. vulgatum extracts showed a potent free radical scavenging activity by 30.4 % at $100\;{\mu}g/mL$ in UVB-irradiated HDFs. UVA induced MMP protein expression was reduced 37.7 % by treatment with O. vulgatum extract, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Taken together, these results suggest that O. vulgatum extract prevents the skin cell damage induced by UV irradiation, and implies that O. vulgatum extract may be useful as a new ingredient for anti-aging cosmetics.

• Journal of Microbiology
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• v.45 no.2
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• pp.158-167
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• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.