• Title/Summary/Keyword: ITS marker

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Transcriptional regulation and mutational analysis of a dctA encoding organic acid transporter protein from Pseudomonas chlororaphis O6.

  • Nam, Hyo-Song;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.100.1-100
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    • 2003
  • A dctA gene encoding a protein with identity to a C4-dicarboxylate/H+ was cloned from a beneficial biocontrol bacterium, P. chororaphis O6. Expression of the dctA was induced in minimal medium by several organic acids and was repressed by glucose. Highest expression was observed in early-log cells grown on fumarate and succinate with decline as cells approached late-log phase. The dctA transcript accumulated weakly when cells were grown on malate but strong expression was observed with benzoate. Expression of the dctA transcript was repressed in early-log cells upon addition of glucose to fumarate, but was detected as the cell culture aged. A dctA-deficient mutant of O6, constructed by marker exchange mutagenesis, did not grow on minimal medium containing succinate, benzoate, or fumarate, and growth on malate was delayed. The dctA mutant and wild type grew equally on glucose. The dctA mutant on cucumber roots in sterilized potting soil was colonized at levels comparable to those of the wild type, but induction level of disease resistance by the mutant against target leaf spot disease was decreased. These results may indicate that the dctA is essential for utilization of certain organic acids and its expression is controlled by the availability of sugars. In addition, the dctA is not essenitial for cucumber root colonization, but important for induction of disease resistance.

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Analysis of Electronic Endoscopic Image of Intramucosal Gastric Carinoma by Using Homoglobin Index (혈색소지수를 이용한 점막내 위암의 전자내시경 영상 분석)

  • Kim Gwang-Ha;Lim Eun-Kyung;Kim Gwang-Baek
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2005.11a
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    • pp.535-541
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    • 2005
  • It has been suggested that the endoscopic color of intramucosal gastric carcinoma is correlated with mucosal vascularity within the carcinomatous tissue. The development of electronic endoscopy has made it possible to quantitatively measure the mucosal hemoglobin volume, using a hemoglobin index. The aim of this study was to make a software program to calculate the hemoglobin index (IHb) and then investigate whether the mucosal IHb determined from the electronic endoscopic data is a useful marker for evaluating the color of intramucosal gastric carcinoma, in particular with regard to its value for discriminating between the histologic type. The mean values of IHb for the carcinoma (IHb-C) and the mean values of IHb for the surrounding non-cancerous mucosa ( IHb-N) were calculated in 75 intestinal-type and 34 diffuse-type gastric carcinomas. Then, we analyzed the ratio of the IHb-C to IHb-N. The mean IHb-C/IHb-N ratio in the intestinal-type carcinoma group was higher than that in the diffuse-type carcinoma group ($1.28{\pm}0.19$ vs. $0.81{\pm}0.18$, respectively, p<0.001). When the cut-off point of the C/N ratio was set at 1.00, the accuracy rate, the sensitivity, the specificity, and the positive and negative predictive values of a C/N ratio below 1.00 for the differential diagnosis of diffuse-type carcinoma from intestinal-type carcinoma were $94.5\%$, $94.1\%$, $94.7\%$, $88.9\%$ and $97.3\%$, respectively. IHb is useful for quantitative measurement of the endoscopic color in intramucosal gastric carcinoma and the IHb-C/IHb-N ratio would be helpful in distinguishing diffuse-type carcinoma from intestinal -type carcinoma.

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A Case of Microscopic Polyangiitis Presented as Pleural Effusion (흉막삼출로 발현된 현미경적 다발혈관염 1예)

  • Shin, Jin-Kyeong;Kwon, Soon-Seog;Park, Ki-Hoon;Lee, Hee-Joung;Kim, Yong-Hyun
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.2
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    • pp.197-202
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    • 2012
  • Microscopic polyangiitis is a necrotizing vasculitis, characterized by inflammation of small vessels (capillaries, venules, and arterioles) with few or no immune deposits. The kidneys are the most commonly affected organs and are involved in 90% of patients, whereas pulmonary involvement occurs in a minority of cases (10% to 30%). In cases of lung disease, diffuse alveolar hemorrhage with pulmonary capillaritis is the most common manifestation. Microscopic polyangiitis is strongly associated with antineutrophil cytoplasmic autoantibody, which is a useful diagnostic serological marker. We report a case of microscopic polyangiitis presented as pleural effusion in a 67-year-old female. Pleural effusions have been reported in some cases previously, but the number of cases were small and their characteristics have not been well described. This report describes characteristic findings of pleural fluid and its histological features in a case of microscopic polyangiitis.

Approaches to Improving Production Efficiencies of Transgenic Animals

  • Tojo, Hideaki
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.7-8
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    • 2000
  • Transgenic animals are very useful for scientific, pharmaceutical, and agricultural purposes. In livestock, transgenic technology has been used forthe genetic alteration of farm animals, the production of human proteins inlarge quantities in the milk of transgenic farm animals, and the generation of animals with organs suitable for xenotransplantation. To date, the transfer of foreign genes into farm animals has been performed mainly by microinjection of DNA into the pronuclei of fertilized eggs. However, the overall success rate of transgenic animals in livestock so far has been disappointingly low, eg., the efficiency is 0∼5% in swine, and less than 1% in sheep and cattle, compared with the rate in mice where 5% microinjected develop into transgenic animals. Recently, McGreath et al. (2000) have succeeded in producing the gene targeted sheep by the use of nuclear transfer from cultured somatic cells transfected with a foreign gene in vitro. However, we may need plenty of time until currently employ this method for gene transfer to farm animals. We have been studying to exploit the method for improving production efficiencies of transgenic animals with emphasis of its application to farm animals. The present paper describes three approaches that we have made in our laboratory to improve production efficiencies of transgenic animals, based on the DNA microinjection method. 1. Co-injection of restriction enzyme with foreign DNA into the pronucleus for elevating production efficiencies of transgenic animals. 2. Efficient selection of transgenic mouse embryos using EGFP as a marker gene. 3. Phenotypes of tansgenic mice expressing WAP/hGH-CAG/EGFP fusion gene produced by selecting transgenic embryos. 4. Efficient site-specific integration of the transgene targeting an endogenous lox like site in early mouse embryos.

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Retrovirus Vector-Mediated Inductional Expression of the Human Lactadherin Gene in Mouse Mammary Epithelial Cells (Mouse Mammary Epithelial Cell에서 Retrovirus Vector를 이용한 Human Lactadherin 유전자의 유도적 발현)

  • 권모선;구본철;정병현;염행철;박창식;김태완
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.15-23
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    • 2003
  • Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of tissue specific and hormonal inducible mouse whey acidic protein (WAP) promote., the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAP promoter was accomplished in the presence of insulin, hydrocortisone and prolactin. Compared to the control (cells cultured with insulin alone), however we observed that the WAP promoter was leaky. These data indicate that luther studies are needed in finding an appropriate promoter other than WAP promoter because of its leakiness.

Investigation of the Protective Effect of Kefir against Isoproterenol Induced Myocardial Infarction in Rats

  • Mert, Handan;Yilmaz, Hikmet;Irak, Kivanc;Yildirim, Serkan;Mert, Nihat
    • Food Science of Animal Resources
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    • v.38 no.2
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    • pp.259-272
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    • 2018
  • This study aims to investigate the protective effects of kefir against myocardial infarction induced by isoproterenol (ISO). The rats were randomly divided into 4 groups, each group consisting of 8 rats. The control group, the kefir group (5 mL/kg/d kefir administered to rats as intra-gastric gavage for 60 d), the ISO group (100 mg/kg ISO was administered to rats, s.c. on 61. and 62. d), and kefir+ISO group (5 mL/kg/d kefir was administered to rats intra gastric gavage for 60 days prior to ISO, 100 mg/kg in two doses on day 61 and 62). 12 h after the last ISO dose, all rats were decapitated and their blood samples were collected. Cardiac tissue was reserved for histopathological examination. creatine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), triglycerides, total cholesterol,very low density lipoprotein (VLDL), low density lipoprotein (LDL), high density lipoprotein (HDL) and glucose were measured by autoanalyzer, whole blood malondialdehyde (MDA), glutathione (GSH) and plasma advanced oxidation protein products (AOPP) levels were measured spectrophotometrically. It was determined that in the group of kefir+ISO, the levels of AST (p<0.001), CK (p<0.001), LDH (p<0.001), MDA (p<0.001) and AOPP (p<0.001) were decreased, while the GSH (p<0.05) increased, compared to ISO group. There were no significant changes in lipid profile and glucose levels between these two groups. In conclusion, by examining cardiac enzymes and histopathological changes in cardiac tissue, it can be concluded that the administration of kefir in myocardial infarction induced by ISO can protect the heart with its antioxidant characteristic and minimize the toxic damage created by ISO.

Studies on Development of Antagonistic Microorganism by Cell Fusion - Biological control of disease - ) (세포융합에 의한 신 길항미생물 육종에 관한 연구 - 목초 병해의 생물학적 방제 -)

  • 최기춘;이영환;전우복
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.15 no.1
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    • pp.1-12
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    • 1995
  • This study was to investigate an effective biological control of forage diseases and provide a basic data and a model in improving variety of antagonistic bacteria, with growth promoting effect on forage, through cell fusion. The results obtained were summarized as follows; 1. The antagonistic himbacterium against soil-borne phathogenic fungi Fusarium oxysporum and Rhizoctonia solani was isolated from continuous cropping himsphere soil of forage, and its biological and physiological characteristics were investigated. This bacterium was identified as Bacillus subrilis and named BS 101. Another strain for cell fusion was Bacillus thur ingiensis ssp. kurstaki HD-I(BT 37669) with insecticidal crystal. 2. The auxotropic mutants of BS 101 and BT 37669 were derived after mutagenesis using N-methyl-N'nitro- Nitrosoguanidine(NTG) to give amino acid requirement marker. n e s e auxotropic mutants of BS 101 and BT 37669 were named BS 1013(his-) and BT 69(asp-), respectively. 3. The best protoplast requirement was obtained using DM 3 medium, containing 5% casamino acid, 1 M $MgCI_2$ and 2% bovine semm albumin, to give Fusant 3, 7 and 8. BT toxin gene was not identified with fusants by Southern blotting. However, SDS-PAGE analysis of strains showed various protein patterns among fusants. 4. From the dark culture experiment, growth of forage in inoculated soil with antagonistic bacteria was delayed than that of non-inoculated soil with antagonistic bacteria in each continuous cropping soil and in each sterilized soil. On the other hand, growth duration of forage was different between continuous cropping soil and sterilized soil. 5. Seed germination of Alfalfa, Italian ryegrass and Orchardgrass were significantly improved by inoculation of antagonistic bacteria(p< 0.05).

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Ethyl Acetate Fraction of Adenophora triphylla var. japonica Inhibits Migration of Lewis Lung Carcinoma Cells by Suppressing Macrophage Polarization toward an M2 Phenotype

  • Park, Shin-Hyung
    • Journal of Pharmacopuncture
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    • v.22 no.4
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    • pp.253-259
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    • 2019
  • Objectives: It is reported that tumor-associated macrophages (TAMs) contribute to cancer progression by promoting tumor growth and metastasis. The purpose of this study is to investigate the effect of different fractions of Adenophora triphylla var. japonica (AT) on the polarization of macrophages into the M2 phenotype, a major phenotype of TAMs. Methods: We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AT. The cytotoxicity of AT in RAW264.7 cells was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 cells were polarized into the M2 phenotype by treatment with interleukin (IL)-4 and IL-13. The expression of M2 macrophage marker genes was detected by reverse transcription polymerase chain reaction (RT-PCR). The phosphorylation level of signal transducer and activator of transcription 6 (STAT6) was investigated by western blot analysis. The migration of Lewis lung carcinoma (LLC) cells was examined by transwell migration assay using conditioned media (CM) collected from RAW264.7 cells as a chemoattractant. Results: Among various fractions of AT, the ethyl acetate fraction of AT (EAT) showed the most significant suppressive effect on the mRNA expression of M2 macrophage markers, including arginase-1, interleukin (IL)-10 and mannose receptor C type 1 (MRC-1), up-regulated by treatment of IL-4 and IL-13. In addition, EAT suppressed the phosphorylation of STAT6, a critical regulator of IL-4 and IL-13-induced M2 macrophage polarization. Finally, the increased migration of Lewis lung carcinoma (LLC) cells by CM from M2-polarized RAW264.7 cells was reduced by CM from RAW264.7 cells co-treated with EAT and M2 polarization inducers. Conclusion: We demonstrated that EAT attenuated cancer cell migration through suppression of macrophage polarization toward the M2 phenotype. Additional preclinical or clinical researches are needed to evaluate its regulatory effects on macrophage polarization and anti-cancer activities.

Influence of Toxoplasma gondii Acute Infection on Cholinesterase Activities of Wistar Rats

  • Tonin, Alexandre Alberto;da Silva, Aleksandro Schafer;Thorstenberg, Maria Luiza;Castilhos, Livia Gelain;Franca, Raqueli Teresinha;Leal, Daniela Bitencourt Rosa;Duarte, Marta Maria Medeiros Frescura;Vogel, Fernanda Silveira Flores;de La Rue, Mario L.;dos Anjos Lopes, Sonia Terezinha
    • Parasites, Hosts and Diseases
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    • v.51 no.4
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    • pp.421-426
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    • 2013
  • Several studies have shown the mechanisms and importance of immune responses against Toxoplasma gondii infection and the notable role of cholinesterases in inflammatory reactions. However, the association between those factors has not yet been investigated. Therefore, the aim of this study was to evaluate the acetylcholinesterase (AChE) activity in blood and lymphocytes and the activity of butyrylcholinesterase (BChE) in serum of rats experimentally infected with T. gondii during the acute phase of infection. For that, an in vivo study was performed with evaluations of AChE and BChE activities on days 5 and 10 post-infection (PI). The activity of AChE in blood was increased on day 5 PI, while in lymphocytes its activity was enhanced on days 5 and 10 PI (P<0.05). No significant difference was observed between groups regarding to the activity of BChE in serum. A positive (P<0.01) correlation was observed between AChE activity and number of lymphocytes. The role of AChE as an inflammatory marker is well known in different pathologies; thus, our results lead to the hypothesis that AChE has an important role in modulation of early immune responses against T. gondii infection.

The Clinical Significance of Serum Ferritin in Pediatric Non-Alcoholic Fatty Liver Disease

  • Na, Ji Hoon;Park, So Won;Kang, Yunkoo;Koh, Hong;Kim, Seung
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.17 no.4
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    • pp.248-256
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    • 2014
  • Purpose: Non-alcoholic fatty liver disease (NAFLD) in children has become an important public health issue because of its high prevalence and severity. Several noninvasive methods for estimating NAFLD are under investigation. We aimed to evaluate the usefulness of serum ferritin as a biomarker of severity of pediatric NAFLD patients. Methods: A total of 64 NAFLD patient were enrolled from Severance Children's Hospital from March 2010 to February 2013. Serum ferritin levels, liver related laboratory tests, liver magnetic resonance imaging (MRI) (2-dimensional [2D] proton density-fat fraction) and NAFLD severity markers were compared between obese group and overweight group. Correlation analyses were performed between serum ferritin and laboratory values including NAFLD severity markers. Results: In obese group, serum ferritin, alanine aminotransferase (ALT), total bilirubin, international normalized ratio (INR), MRI 2D proton density-fat fraction, aspartate aminotransferase (AST) to platelet ratio index (APRI) and fibrosis- 4 (FIB-4) (an index score calculated from platelet count, ALT, AST and age) were significantly higher than those of overweight group. NAFLD severity markers, APRI and FIB-4, and liver specific important laboratory values, AST, ALT, INR, cholesterol, triglyceride and low density lipoprotein show significant correlation with serum ferritin in NAFLD patients. Conclusion: Serum ferritin concentrations could be a candidate of useful severity marker in the pediatric NAFLD patients.