Patients with head and neck cancer are treated with therapeutic irradiation, which can result in irreversible salivary gland dysfunction. Because there is no complete cure for such patients, stem cell therapy is an emerging alternative for functional restoration of salivary glands. In this study, we investigated in vitro characteristics of primarily isolated epithelial cells from human salivary gland (Epi-SGs) and in vivo formation of acini-like structures by Epi-SGs. Primarily isolated Epi-SGs showed typical epithelial cell-like morphology and expressed E-cadherin but not N-cadherin. Epi-SGs expressed epithelial stem cell (EpiSC) and embryonic stem cell (ESC) markers. During long-term culture, the expression of EpiSC and ESC markers was highly detected and maintained within the core population with small size and low cytoplasmic complexity. The core population expressed cytokeratin 7 and cytokeratin 14, known as duct markers indicating that Epi-SGs might be originated from the duct. When Epi-SGs were transplanted in vivo with Matrigel, acini-like structures were readily formed at 4 days after transplantation and they were maintained at 7 days after transplantation. Taken together, our data suggested that Epi-SGs might contain stem cells which were positive for EpiSC and ESC markers, and Epi-SGs might contribute to the regeneration of acini-like structures in vivo. We expect that Epi-SGs will be useful source for the functional restoration of damaged salivary gland.
Kim, Ji-Hwan;Kim, Goo-Young;Nam, Min-Kyung;Kim, Sang-Soo;Rhim, Hyang-Shuk
Korean Journal of Microbiology
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v.45
no.4
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pp.291-299
/
2009
To elucidate HtrA3's functional roles in the HtrA3 mediated cellular processes, it is necessary to investigate its biochemical characteristics. In the present study, we constructed the plasmids encoding putative mature HtrA3 proteins (M1-HtrA3 and M2-HtrA3) based on the putative maturation sites of highly homologous HtrA1 and mouse HtrA3. We used the pGEX bacterial expression system to develop a simple and rapid purification for the recombinant HtrA3 protein. Although yields of the mature HtrA3 proteins were slightly low as 10~50 ${\mu}g$/L, the amounts and purity of M1- and M2-HtrA3 were enough to investigate their proteolytic activities. The putative mature HtrA3 proteins have proteolytic activity which could cleave $\beta$-casein as an exogenous substrate. We compared the proteolytic activity between the HtrA family, HtrA1, HtrA2, and HtrA3. The cleavage activity of HtrA3 and HtrA2 were 2 folds higher than that of HtrA1, respectively. Our study provides a method for generating useful reagents to identify natural substrates of HtrA3 in the further studies.
This study was conducted to examine community residents' knowledge level and related factor on electronic wave in order to provide basic data for development of education and publicity program. 2,000 people, who lived in five big cities and five small and medium cities, were selected ad subjects of this study. The data were collected from May 1, 200 I to August 31, 2001. The results of this study are as follows. According to the average knowledge level of harmful affect of electronic wave on health in general characteristics, female was higher(37.40 ± 5.24 points) than male; ‘forties’ was highest(37.77 ± 5.69 points); ‘married spouse’ was high(36.84 ± 5.59 points); ‘living in small-ta-medium city’ was high(36.84 ± 5.32 points). ‘university graduate’ was highest(37.41 ± 5.32 points) in education level, ‘middle class’ was high(36.61 ± 4.96 points) in economic status, ‘professional technician’ was higher(36.68 ± 6.55 points) than other occupations in occupational type. According to the knowledge level of harmful affect of electronic wave on health in health condition by self-judgment, ‘good health condition’ was highest(36.77 ± 4.99 points). In the case of the knowledge level of those who visited medical institutions for last one year, ‘never visited’ was highest(37.19 ± 5.02 points). In the kind of medical institutions, ‘those who visited general hospital’ was highest(36.58 ± 5.63 points). In the way of knowledge obtainments of electronic wave through education and publicity media, ‘school education’ was highest(37.55 ± 5.19 points). According to the score of awareness level of disease incidence related to electronic wave, allergy and erethism was highest(57.8 points on the basis of 100 points). It appeared in order of leukemia, skin disease or skin cancer, dementia, various cancers, cataract, and brain tumor. The variables which significantly influenced knowledge level of harm of electronic wave were knowledge obtainments of electronic wave, age, economic status, daily TV watching period, sex, period of daily cellular phone use, period of working with computer, and daily VTR watching period. The knowledge of community residents concerning harmful affect of electronic wave on health is needed because people's opportunity of exposing to electronic wave is increasing. Especially, it is the demands of the times to provide information on knowledge of each equipment which generate electronic wave. The government, the product manufacturing companies, related social organizations, and education institutions must make efforts to develop the education program which is needed to make people have right knowledge and attitude.
Production of a high viscosity exoploysaccharide, methylan, by Methylobacterium organophilum from methanol was carried out in fed-batch cultures and the rheological properties of methylan fermentation broth were studied. Bacterial biomass showed little influence on viscosity, but the accumulation of methylan caused the increase of viscosity. With proceeding fermention, the viscosity at the same concentration of methylan was significantly increased and methylan solution showed slightly higher pseudoplasticity. The composition changes of methylan were investigated at various fermentation times. Contents of total sugar, reducing sugar and methylan were decreased but contents of acids(pyruvic acid, uronic acid and acetic acid) were increased with the culture time. It was considered that the increased content of acids resulted in the increase of the hyrodynamic domain in the solution due to charge repulsion. Consequently, the solution viscosity increased in propotion to the acids contents of methylan. Cell growth and methylan production were severely decreased by the limitation of dissolved oxygen. However, the cellular activity for methylan production was almost constant regardless of the level of dissolved oxygen. As a result, the high speed of agitation increased the methylan production, the specific production rate of methylan, and the methylan yield of the cell.
The purpose of this work is to develope sodium-containing nonstoichiometric apatitic coatings on solid substrate. The apatitic coatings prepared at different concentrations of sodium salt indicated that the presence of sodium ions exerted significant effects on the physicochemical properties of the apatitic coating including surface morphology, chemical state, and Ca/P ratio. The variation of these properties was sustained up to 0.01 mM of sodium ion concentration. The ratio of calcium to phosphorus was varied from 2.18 to 2.03 which indicated the apatitic coating prepared in this study was calcium-rich nonstoichiometric apatite. The structure of all the samples appeared to be low crystalline. In the presence of sodium ion within the apaptitic coating, the adhesion of human osteoblast-like SaOS-2 cells was significantly promoted. On the other hand, the proliferation of the cells on the apatitic coatings was decreased with the increase of sodium ions. This reverse response of SaOS-2 cells indicates that the interaction between SaOS-2 and apatitic surface triggered considerable changes in intracellular mechanisms including cellular signal transductions.
Oxidative stress leads to the induction of cellular oxidative damage, which may cause adverse modifications of DNA, proteins, and lipids. The production of reactive species during oxidative stress contributes to the pathogenesis of many diseases. Antioxidant defenses can neutralize reactive oxygen species and protect against oxidative damage. The aim of this study was to assess the antioxidant status and the degree of DNA damage in Korean young adults using glutathione s-transferase (GST) polymorphisms. The GSTM1 and GSTT1 genotypes were characterized in 245 healthy young adults by smoking status, and their oxidative DNA damage in lymphocytes and antioxidant status were assessed by GST genotype. General characteristics were investigated by simple questionnaire. From the blood of the subjects, GST genotypes; degree of DNA damage in lymphocytes; the erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase; plasma concentrations of total peroxyl radical-trapping potential (TRAP), vitamin C, ${\alpha}$- and ${\gamma}$-tocopherol, ${\alpha}$- and ${\beta}$-carotene and cryptoxanthin, as well as plasma lipid profiles, conjugated diene (CD), GOT, and GPT were analyzed. Of the 245 subjects studied, 23.2% were GSTM1 wild genotypes and 33.4% were GSTT1 wild genotype. No difference in erythrocyte activities of superoxide dismutase, catalase, or glutathione peroxidase, and the plasma TRAP level, CD, GOT, and GPT levels were observed between smokers and non-smokers categorized by GSTM1 or GSTT1 genotype. Plasma levels of ${\alpha}$- and ${\gamma}$-tocopherol increased significantly in smokers with the GSTT1 wild genotype (p < 0.05); however, plasma level of ${\alpha}$-carotene decreased significantly in non-smokers with the GSTM1 wild genotype (p < 0.05). DNA damage assessed by the Comet assay was significantly higher in non-smokers with the GSTM1 genotype; whereas DNA damage was significantly lower in non-smokers with the GSTT1 genotype. Total cholesterol and LDL cholesterol levels were significantly higher in non-smokers with the GSTT1 genotype than those with the GSTT1 wild genotype (p < 0.05). In conclusion, the GSTM1 genotype or the GSTT1 wild genotype in non-smokers aggravated their antioxidant status through DNA damage of lymphocytes; however, the GSTT1 wild type in non-smokers had normal plasma total cholesterol and LDL-cholesterol levels. This finding confirms that GST polymorphisms could be an important determinant of antioxidant status and plasma lipid profiles in non-smoking young adults. Further study is necessary to clarify the antioxidant status and/or lipid profiles of smokers with the GST polymorphism and to conduct a study with significantly more subjects.
Demineralized boneparticle (DBP) has been widely used as and a powerful promoter of new bone growth. In this study, DBP sponges were chemically crosslinked and characterized for the potential application of tissue engineered scaffolds. The DBP sponges prepared by crosslinking with EDC. 0.1, 0.2 or 0.3% pepsin was applied to DBP dissolved in 3% (v/v) acetic acid aqueous solution for 48 hrs. The prepared sponges were crosslinked by 1, 5, 10, 50 or 100 mM of EDC solution concentration and then were lyophilized. The DBP sponges were characterized by SEM, FT-IR and DSC and analyzed in terms of their porosity and water absorption ability. The cellular viability and proliferation were assayed by MTT assay. Our investigation revealed that 0.2$\sim$0.3% of pepsin and 50$\sim$100 mM of EDC produced DBP sponges with good physical characteristics. In conclusion, DBP sponge prepared under these conditions is potentially useful for the applications of tissue construction.
Jung, Won Kwon;Kim, Young Soo;Choi, Jin Kook;Kim, Seung-Han;Jang, Myeong-Hwan;Kwon, Tae Lyong;Jeon, Yongho
Research in Plant Disease
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v.24
no.4
/
pp.332-338
/
2018
In August of 2011, a wilting disease of ginseng was observed at Bongwha, Gyeongbuk province, Korea. Affected plants initially show withering symptoms on leaves of ginseng. As the disease progresses, withering leaves spread downward, eventually encompassing the whole plant. Leaves lose vigor but remain pale green. Symptoms of roots were brown, and soft rots characterized by moist and watery decay of the whole ginseng root, which initiated as small brown, water-soaked lesions of hairy roots and enlarged to the entire roots. The causal organism isolated from the infected roots was identified as Serratia plymuthica based on its physiological and biochemical characteristics, by cellular fatty acid composition (GC-FAME), the utilization of carbon sources (BioLog System), and 16S rRNA sequence of the isolated bacterium were 99% homologous to those of Serratia plymuthica strains. Artificial inoculation of the bacterium produced the same brown or soft rot symptoms on the ginseng roots, from which the same bacterium was isolated. This is the first report of bacterial root rot caused by the Serratia plymuthica in ginseng in Korea. Serratia plymuthica has been used as antagonistic microorganism for biological control on several crop plants. But it was proved pathogen of ginseng at humid condition in this study.
Ahn, Jae-Sung;Park, Kyung Sun;Park, Jongha;Chung, Hyun Chul;Park, Hojong;Park, Sang Jun;Cho, Hong Rae;Lee, Jong Soo
Korean Journal of Transplantation
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v.31
no.4
/
pp.182-192
/
2017
Background: In recent years, introduction of novel immunosuppressive agents and its proper implementation for clinical practice have contributed to improving clinical outcomes of kidney transplantation (KT). Here, we report clinical outcomes of KTs and related risk factors. Methods: From July 1998 to June 2016, 354 KTs (182 from living and 172 from deceased donors) have been performed at Ulsan University Hospital. We retrospectively reviewed the clinical characteristics and outcomes of KT recipients, then estimated graft and patient survival rate were estimated and analyzed risk factors using Cox-regression. Results: The median follow-up period was 53 months (range; 3 to 220 months). The mean ages of recipients and donors were 45.0 years (SD, 12.5) and 44.7 years (SD, 13.6) years, respectively. During follow-up, 18 grafts were lost and 5- and 10-year death-censored graft survival was 96.7% and 91.5%, respectively. Biopsy-proven acute rejection (BPAR) occurred in 71 patients (55 cases of acute cellular rejection and 16 of antibody-mediated rejection). Cox-regression analysis showed that BPAR was a risk factor related to graft loss (hazard ratio [HR], 14.38; 95% confidence interval [CI], 3.79 to 54.53; P<0.001). In addition, 15 patients died, and the 5- and 10-year patient survival was 97.2% and 91.9%, respectively. Age ≥60 years (HR, 6.03; 95% CI, 1.12 to 32.61; P=0.037) and diabetes (HR, 6.18; 95% CI, 1.35 to 28.22; P=0.019) were significantly related to patient survival. Conclusions: We experienced excellent clinical outcomes of KT in terms of graft failure and patient survival despite the relatively high proportion of deceased donors. Long-term and short-term clinical outcomes have improved in the last two decades.
Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.
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