• Journal of Genetic Medicine
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• v.4 no.1
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• pp.45-52
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• 2007

Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

• Journal of Life Science
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• v.28 no.11
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• pp.1339-1346
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• 2018

In Korea, edible mushrooms are produced largely on commercial artificial media, so the annual production of spent mushroom substrate (SMS), as a by-product of the mushroom industry, is estimated at over 200 million tons. This SMS is assumed to contain abundant fungal mycelia and pre-fruiting bodies, as well as various nutritive and bioactive compounds that are presently discarded. This study examined the physico-chemical, nutritional, and enzymatic characteristics of uninoculated sterilized medium (USM) and SMS of shiitake mushrooms with the aim of developing a high-value added product from SMS. The contents of crude protein, crude lipid, and ash were higher after the third SMS harvest ($SMS-A-3^{rd}$) than in USM or $SMS-A-1^{st}$. The contents of Ca, Mg, and P in $SMS-A-3^{rd}$ were 2.95, 2.35, and 2.1-fold higher compared than in USM. No As or Cd was detected in USM or SMS. The pH, Brix, and acidity were 4.6, 20.0, and 1.4, respectively in $SMS-A-3^{rd}$, but 5.6, 6.0, and 0.0, respectively, in USM. These results suggest a highly active production of soluble components and organic acids in $SMS-A-3^{rd}$. The distinct color differences noted for USM, $SMS-A-1^{st}$, and $SMS-A-3^{rd}$ could be used as a mycelial growth indicator. Enzyme activity assays using the APIZYM system showed that SMS is a potent source of hydrolysis-related enzymes, especially esterase (C4) and ${\beta}$-glucuronidase. Our results suggested that the SMS of shiitake has a high potential for use in environmental, agricultural, and stock-breeding industries, for example, as active ingredients for sewage treatment, waste-polymer degradation, and feed additives.

• International Journal of Oral Biology
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• v.32 no.1
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• pp.35-43
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• 2007

Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated ${\beta},-galactosidase$, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and $p16^{INK4A}$ protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although $p27^{Kip1}$ and $p15^{INK4B}$, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin ${\alpha}2$, ${\alpha}v$, and ${\beta}1$. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that $p16^{INK4A}/RB$ might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.

• Journal of Life Science
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• v.8 no.3
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• pp.263-271
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• 1998

One of the better-characterized transcription factor of E. coli is the cAMP receptor protein(CRP) and the CRP binds cAMP and DNA. The cAMP-CRP complex is involved in regulation of many genes at bacteria. The cAMP-CRP regulatory element represents, in some respects, a global regulatory network. The aim of this work was to study the structure and the mechanisms controlling the expression of CRP in Serratia marcescens. We have been get 5 different clones from Serratia which stimulated the cells to use maltose as a sole carbon source in E. coli TP2139. The crp gene clone, pCKB12, was confirmed by Southern hybridization with E. coli crp gene. The location of the crp gene was determined by construction subclones carrying various portions of pCKB12. To investigate the potential role of CRP in E. coli, lacZ fused plasmids were constructed and investigated the ${\beta}$-galactosidase activity of the fused plasmid. The Serratiamarcescens cAMP receptor protein can substitute the E. coli CRP in transcriptional activation at the lacZ gene. These results suggest that Serratia marcescens cAMP receptor protein complex functions to regulate several promoters in E. coli.

• KSBB Journal
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• v.16 no.5
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• pp.486-492
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• 2001

The antimutagenic mechanism of the fraction III(RG III)separated from the water extract of Rehmannia glutionosa was investigated by Escherichia. coli GW and B/r strains. RG-III treatment did not affect the ${\beta}$-galactosidase activity E. coli GW-1060, 1106, 1107 and 1105. These results indicated that RG-III did not induce RecA protein amplification and did not also prevent the proteolytic cleavage of LexA. The bio-antimutagenicity and survival effect of RG-III on 4-nitroquinoline 1-oxide(4NQO), N-methyl-N-nitor-N\`-nitrosoguanidine(MNING) were investigate by E. coli B/r strains with have different pathway of DNA repai. RG-III slightly increased the survival of 4NQO-treated WP2, WP2s, WP67, CM561, CM611 cells, but the reactivation of survival cannot ve explained by the repair mode. RG-III caused the decrease of mutagenicity and lethality treated with MNNG in ZA159 despite of the increase in WP2, WP2s, WP67, CW561, CM611. Compared with bio-antimutagenic effects of RG-III on 4NQO, greatly increased antimutagenic effects of RG-III were observed with all the E. coli B/r strains tested, but less active in ZA159. These results suggest that RG-III was identified as a blocking agent for preventing the 4NQO induced mutagenesis, and may act as chl-products.

• Environmental Analysis Health and Toxicology
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• v.15 no.4
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• pp.123-130
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• 2000

2-Bromopropane, important industrial chemical, specially in electronic industry at Yangsan in Korea has been reported to cause amenorrhea for female and azoospermia, oligozoospermia or reduced sperm motility for male. 2-BP was investigated through 21 days of repeated dose in male Sprague-Dawley rats. The dose levels per body weight were 0 (control), 250,500 and 1,000 mg/kg. 2-BP dissolved in vehicle olive oil was injected into the intraperitoneum 6 times per week for 3 weeks, but 1,000 mg/kg dose group was 2 weeks because of serious illness. Male rats showed significant decreases in body weight and right and left testis showed typical weight losses depending on the 2-BP. The number of white blood cell and red blood cell , percentage of monocytes, and hemoglobin decreased significantly in high dose (P< 0.05). Red cell volume distribution width increased significantly in the high dose (P< 0.05). Histopathological findings of testes showed a decrease of spermatogenic cells, exfoliation of spermatid and spermatocyte, vacuolization of Sertoli cells and hyperplasia of Leydig cells. Protein band density between 113,000 dalton ($\beta$-galactosidase) and 53,900 dalton (ovalbumin) has decreased in 250 mg/kg dose group, but it has gradually increased to the higher density in 1,000 mg/kg dose group than in control group.

• Journal of Food Hygiene and Safety
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• v.32 no.3
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• pp.243-248
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• 2017

Intracellular and extracellular oxidative stress initiated by reactive oxygen species (ROS) causes skin aging, which is characterized by wrinkles and atypical pigmentation. Use of antioxidant is an effective approach to prevent symptoms related to ROS-induced aging of the skin. Therefore, the antioxidant and anti-aging effect of Castanea crenata Siebold & Zucc. extracts (LCE) was investigated in this study. The LCE markedly reduced the hydrogen peroxide-induced cell damage, intracellular ROS, and oxidative stress-induced senescence in human dermal fibroblasts (HDFs). These results indicate that LCE might have beneficial effects on oxidative stress-induced damage and thus reduce skin aging.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6191-6196
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• 2012

Aims and Background: Traditional chemotherapy strategies for human leukemia commonly use drugs based on cytotoxicity to eradicate cancer cells. One predicament is that substantial damage to normal tissues is likely to occur in the course of standard treatments. Obviously, it is urgent to explore therapies that can effectively eliminate malignant cells without affecting normal cells. Our previous studies indicated that ginsenoside $Rg_1$ ($Rg_1$), a major active pharmacological ingredient of ginseng, could delay normal hematopoietic stem cell senescence. However, whether $Rg_1$ can induce cancer cell senescence is still unclear. Methods: In the current study, human leukemia K562 cells were subjected to $Rg_1$ exposure. The optimal drug concentration and duration with K562 cells was obtained by MTT colorimetric test. Effects of $Rg_1$ on cell cycle were analyzed using flow cytometry and by SA-${\beta}$-Gal staining. Colony-forming ability was measured by colony-assay. Telomere lengths were assessed by Southern blotting and expression of senescence-associated proteins P21, P16 and RB by Western blotting. Ultrastructural morphology changes were observed by transmission electron microscopy. Results: K562 cells demonstrated a maximum proliferation inhibition rate with an $Rg_1$ concentration of $20{\mu}\;mol{\cdot}L^{-1}$ for 48h, the cells exhibiting dramatic morphological alterations including an enlarged and flat cellular morphology, larger mitochondria and increased number of lysosomes. Senescence associated-${\beta}$-galactosidase (SA-${\beta}$-Gal) activity was increased. K562 cells also had decreased ability for colony formation, and shortened telomere length as well as reduction of proliferating potential and arrestin $G_2$/M phase after $Rg_1$ interaction. The senescence associated proteins P21, P16 and RB were significantly up-regulated. Conclusion: Ginsenoside $Rg_1$ can induce a state of senescence in human leukemia K562 cells, which is associated with p21-Rb and p16-Rb pathways.

• Journal of Dairy Science and Biotechnology
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• v.31 no.2
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• pp.99-108
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• 2013

Kefir is a unique fermented dairy product resulting from combined lactic acid and alcoholic fermentation of lactose in milk. Kefir is produced by the microbial activity of kefir grains. Kefir has numerous health benefits such as the reduction of cholesterol levels, stimulation of the immune system, antimutagenic and anticarcinogenic properties, and improvements of lactose intolerance. Furthermore, kefir is excellent as both a dietetic beverage and for protection against various diseases in small babies. Therefore, kefir has recently been regarded as an important functional dairy food. To date, the majority of research on kefir has focused on the applications of functional kefir using advanced biotechnology methods. The purpose of this review article is to facilitate the recognition of kefir as a novel functional food.

• Journal of Dairy Science and Biotechnology
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• v.31 no.2
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• pp.143-152
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• 2013

Aminobutyric acid (GABA), a major inhibitory neurotransmitter in the central nervous system of animals, has several physiological effects including anti-hypertensive, diuretic, tranquilizing, and anti-stress properties, in humans. The purpose of this study was to investigate Lactobacillus plantarum K74, which was isolated from kimchi and selected as a strain with a high ability to produce GABA, to develop a new starter culture for fermented milk production. L. plantarum K74 produced $134.52{\mu}g/mL$ GABA in MRS broth containing 1% MSG, $212.27{\mu}g/mL$ GABA in MRS broth containing 2% MSG, and $234.63{\mu}g/mL$ GABA in MRS broth containing 3% MSG. The optimum growth temperature of L. plantarum K74 was $34^{\circ}C$, reaching a pH of 4.4 after 18 hours of growth. L. plantarum K74 was most sensitive to novobiocin out of 16 different antibiotics tested, and was most resistant to kanamycin and polymyxin B. L. plantarum K74 did not produce ${\beta}$-glucuronidase, a carcinogenic enzyme, and was comparatively tolerant to bile juice and low pH. Furthermore, it displayed resistance to Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus at rates of 54.9%, 46.3%, and 0.7%, respectively.