• Title/Summary/Keyword: IS-PCR

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Immune Regulatory Function of Dendritic Cells Expressing Indoleamine 2,3-Dioxygenase in Orally Tolerance to Type II Collagen-induced Animal Model (제2형 콜라겐 경구관용 유도 동물모델에서 수지상 세포의 Indoleamine 2,3-dioxygenase의 의존성 관절염 항원 특이 T세포 증식반응 제어 연구)

  • Park, Min-Jung;Min, So-Youn;Park, Kyoung-Su;Cho, Mi-La;CHo, Young-Gyu;Min, Jun-Ki;Yoon, Chong-Hyeon;Park, Sung-Hwa;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.5 no.4
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    • pp.221-231
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    • 2005
  • Background: Immune regulatory dendritic cells (DCs) play an important role in maintaining self-tolerance. Recent evidences demonstrate that DCs expressing indoleamine 2,3-dioxygenase (IDO), which is involved in tryptophan catabolism, play an important role in immunoregulation and tolerance and induce T cell apoptosis. This study was devised to examine the role of IDO in the oral tolerance induction in collagen-induced arthritis (CIA) mouse model. Methods: Beginning 2 weeks before immunization, CII was fed six times to DBA/1 mice and the effect on arthritis was assessed. In tolerized mice, $CD11c^+$ DCs were isolated and stimulated with CII, IFN-${\gamma}$, and LPS with or without IDO inhibitor, 1-methyl-DL-tryptophan (1-MT) and IDO expression by $CD11c^+$ DCs was analyzed using FACS and RT-PCR. The expression of IDO, MHC II, CD80, and CD86 by $CD11c^+$ DCs were examined using confocal microscopy. Regulatory effect of $CD11c^+$ DCs on Ag-specific T cell proliferative response to CII was examined by mixed lymphocyte reaction (MLR) with or without 1-MT. Results: The proportion of IDO-expressing $CD11c^+$ DCs was slightly higher in tolerized mice than in CIA mice and significantly increased after stimulation with CII, IFN-${\gamma}$, and LPS in an IDO-dependent manner. On confocal microscopic examination, the expression of IDO was higher and those of MHC II and CD86 were lower in CD11c + DCs from tolerized mice compared to those from CIA mice. On MLR, $CD11c^+$ DCs from tolerized mice inhibited T cell proliferative response to CII in an IDO-dependent manner. Conclusion: Enhanced IDO expression by $CD11c^+$ DCs from tolerized mice may contribute to the regulation of proliferative response of CII-reactive T cells and could be involved in the induction of oral tolerance to CII.

Biosynthesis of polyhydroxyalkanoate by mixed microbial cultures from hydrolysate of waste activated sludge (혼합미생물배양체를 이용한 폐활성슬러지 가용화 산물로부터 polyhydroxyalkanoate 생합성)

  • Park, Taejun;Yoo, Young Jae;Jung, Dong Hoon;Lee, Sun Hee;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.200-207
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    • 2017
  • A new approach to the solubilization of waste activated sludge (WAS) using alginate-quaternary ammonium complex beads was investigated under controlled mild alkaline conditions. The complex beads were prepared by the reaction of sodium alginate (SA) with 3-(trimethoxysilyl)propyl-octadecyldimethylammonium chloride (TSA) in acid solution, followed by crosslinking with $CaCl_2$. Treatment of WAS with SA-TSA complex beads was effective for enhancing the efficacy of WAS solubilization. The highest value of soluble chemical oxygen demand (SCOD) concentration (3,900 mg/L) was achieved after 10 days of treatment with 30% (v/v) SA-TSA complex beads. The WAS solubilization efficacy of the complex beads was also evaluated by estimating the concentrations of volatile fatty acids (VFAs). The maximum value of VFAs was 2,961 mg/L, and the overall proportions of VFAs were more than 75% of SCOD. The main components of VFAs were acetic, propionic, iso-butyric, and butyric acids. These results suggest that SA-TSA complex beads might be useful for enhancing the solubilization of WAS. The potential use of VFAs as the external carbon substrate for the production of polyhydroxyalkanoate (PHA) by a mixed microbial culture (MMC) was also examined. The enrichment of PHA-accumulating MMC could be achieved by periodic feeding of VFAs generated from WAS in a sequencing batch reactor. The composition of PHA synthesized from VFAs mainly consisted of 3-hydroxybutyrate. The maximum PHA content accounted for 25.9% of dry cell weight. PHA production by this process is considered to be promising since it has a doubly beneficial effect on the environment by reducing the amount of WAS and concomitantly producing an eco-friendly biopolymer.

A Microbiological Analysis and Hazard Factor Evaluation of Food Utensils and Fixtures of Food Service Operations in Schools (학교 집단급식소 내 식기류 및 집기류의 미생물학적 분석 및 위해요인 평가)

  • Park, Sung-Jun;Hong, Sung-Ho;Lee, Anne Ha-Young;Kim, Cheol-Ju;Kim, Su-Jin;Kim, Sung-Kyoon;Ko, Gwang-Pyo
    • Journal of Environmental Health Sciences
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    • v.37 no.5
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    • pp.376-386
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    • 2011
  • Objectives: The aim of this study was to evaluate the microbial hazards posed by food utensils and fixtures in food service operations at selected middle and high schools located in Seoul, Korea. Methods: We collected 200 samples of utensils and fixtures including cups, spoons/chopsticks, food trays and tables from five different schools in Seoul. Target microorganisms of this study were divided into two groups: total bacterial count and total coliform as indicators of microbial contamination and Bacillus cereus and Staphylococcus aureus as pathogens of food poisoning. We used selective media to quantify microbial concentration and 16S rRNA PCR assay for qualitative analysis. In addition, intensive interviews with nutritionists were conducted and observations were made to identify factors that may affect microbial contamination. Logistic regression analysis was employed to examine the relationship between the microbial concentration and operation characteristics of each operation. Results: The level of microbial concentration in school B and C were significantly lower than in school A, D and E (p<0.05). Some samples from school A, D and E showed over 3.4 log CFU/100 $cm^2$ (total bacterial count) and 1.0 log CFU/100 $cm^2$ (total coliform), which requires immediate hygienic action. The number of customers per staff member, periodicity of hygiene education for staff and daily operation time of sterilizers were also found to be important factors related with the microbial contamination of food service operations. Conclusions: These results suggested that not only a HACCP (Hazard Analysis and Critical Control Point) approach, but also efforts to assess internal risk factors within operations be needed to reduce the microbial contamination of food utensils and fixtures. This study is expected to provide preliminary data for assessing microbial hazards in food service operations.

The Effect of Tissue Plasminogen Activator on TGF-${\beta}1$ Pre-Treated Human Mesothelial Cell Line (TGF-${\beta}1$으로 자극한 사람중피세포주에서 조직플라스미노겐 활성제가 미치는 영향)

  • Lee, Jung-Lim;Jeon, Soo-Jin;Yoo, Young-Choon;Kim, Ji-Hye;Lee, Yu-Mi;Kwon, Sun-Jung;Son, Ji-Woong;Choi, Eu-Gene;Na, Moon-Jun
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.5
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    • pp.405-415
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    • 2011
  • Background: In an effort to find alternative therapeutic agents to prevent excessive fibrosis as a sequela to complicated parapneumonic effusion or empyema, we examined the effect of tissue plasminogen activator (tPA) as a fibrinolytic agent combined with talc or transforming growth factor (TGF)-${\beta}1$ in a human pleural mesothelial cell line, MeT-5A. Methods: MeT-5A cells were stimulated with various doses of talc, doxycycline or TGF-${\beta}1$ for 24 h and then were treated with tPA for an additional 24 h. Cell viability was measured by MTT assay. The production of interleukin (IL)-8 and vascular endothelial growth factor (VEGF) in the culture supernatants was measured by ELISA. Real-time PCR was carried out for measurement of type I collagen mRNA. Results: MeT-5A cells treated with talc showed a dose-dependent increase in production of IL-8. Talc also increased production of type I collagen mRNA at low doses, but talc did not influence the induction of VEGF. Addition of tPA to talc-stimulated cells showed further increases in the production of IL-8, but tPA did not influence the production of VEGF or type I collagen mRNA. TGF-${\beta}1$ increased the production of both VEGF and collagen type I mRNA, both of which were effectively inhibited by additional tPA treatment in MeT-5A cells. Conclusion: TGF-${\beta}1$ is a potent inducer of collagen synthesis without induction of IL-8 in MeT-5A cells. Addition of tPA after TGF-${\beta}1$ stimulation inhibited further fibrosis by direct inhibition of collagen mRNA synthesis as well as by inhibition of VEGF production.

Clinical Significance of Measuring Levels of CEA, CA19-9 in Peritoneal Washing Fluid in Patients with Gastric Cancer (위암 환자에서 복강 세척액의 CEA, CA19-9 측정의 임상적 의의)

  • Shim, Gyu-Beom;Park, Ji-Hun;Koo, Tea-Young;Min, Hyun-Sik
    • Journal of Gastric Cancer
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    • v.6 no.3
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    • pp.125-131
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    • 2006
  • Purpose: Free cancer cells exfoliated from cancer-invaded serosa contribute to peritoneal dissemination, the most frequent pattern of recurrence in patients with gastric cancer. To detect free cancer cells, CEA and CA19-9 were introduced as the markers of gastric cancer, and many methods, such as cytology, immunoassay, and reverse transcription polymerase chain reaction (RT-PCR), exist for detecting them. The aim of this study is to define the clinical significance of using immunoassay to measure the levels of CEA and CA19-9 in the peritoneal washings in patients with gastric cancer. Materials and Methods: The peritoneal washing fluids were obtained from 130 patients with gastric cancer who received a curative gastrectomy, palliative gastrectomy or open and closure. The pCEA and pCA19-9 levels were measured by using immunoassay and cytology. The results were compared with the clinicopathological data. Results: The pCEA and pCA19-9 levels were correlated with tumor invasion, lymph-node metastasis, and stage (P<0.05). Conclusion: A correlation was found between elevated pCEA and pCA19-9 levels measured by immunoassay and the TNM stage. Therefore, a combined pCEA and pCA19-9 assay could be a sensitive detector of peritoneal dissemination, as well as a predictor of postoperative prognosis. pCEA and pCA19-9 may also determine the adjuvant management strategy.

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Effect of Enamel Matrix Drivatives application on the expression of PDLs17, PDLs22 of cultured human periodontal ligament cells in vitro (Enamel Matrix Derivatives가 사람 치주인대 세포의 특이유전자인 PDLs17, PDLs22의 발현에 끼치는 효과)

  • Han, Geun-A;Jang, Hyun-seon;Kok, Jung-Ki;Park, Ju-Chol;Kim, Heoung-Jung;Kim, Jung-Gwan;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.333-344
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    • 2004
  • The enamel matrix derivative (EMD) has been recently used in the periodontal regenerative techniques. The present study was established to investigate the influence of EMD on human periodontal ligament cells using expression of mRNA of periodontal ligament specific gene (PDLs)17, PDLs22, type I collagen when EMD applied to periodontal ligament cells. Periodontal ligament cells were obtained from a healthy periodontium and cultured in Dulbecco's modified Eagle's medium (DMEM) plus 10% fetal bovine serum and ${\beta}-glycerophosphate$ with ascorbic acid. Test groups were two; One adds EMD in culture media and another added EMD and Dexamethasone (DEX) in culture media. Positive control group added DEX in culture media, and negative control group adds niether of EMD nor DEX. $Emdogain^{(R)}$ (Biora, Sweden, 30 mg/ml) was diluted by 75 ${\mu}g/ml$ concentration to culture media. For reverse transcription-polymerase chain reaction (RT-PCR), total RNA isolated on days 0, 7, 14 and 21. mRNA of PDLs17 was expressed on days 14 and 21 in EMD or DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group, the other side, expressed on days 21 in negative control group. mRNA of PDLs22 expressed on days 7, 14 and 21 in EMD group, and expressed on days 14 and 21 in DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group. Negative control group expressed on days 14 and 21. Type I collagen was expressed on all days and all groups. These results indicate that EMD promotes differentiation of periodontal ligament cells, and this is considered to offer basis that can apply EMD to periodontal tissue regeneration technique.

Oxidative Stress Inhibitory Effects of Low Temperature-Aged Garlic (Allium sativum L.) Extracts through Free Radical Scavenging Activity (저온숙성마늘의 라디칼 소거 활성을 통한 산화스트레스 억제 효과)

  • Hwang, Kyung-A;Kim, Ga Ram;Hwang, Yu-Jin;Hwang, In-Guk;Song, Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.27-34
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    • 2016
  • Garlic has drawn attention as a food material for its anti-oxidative and anti-inflammatory properties as well as for prevention and treatment of cancer. In order to increase efficiency, various aging methods for garlic have been attempted. In particular, thermally processed garlic is known to have higher biological activities due to its various chemical changes during heat treatment. Therefore, in this study, we investigated the anti-oxidative effects of garlic extracts aged at low temperature ($60{\sim}70^{\circ}C$). In the results, 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis (3-ethylbenzo-thiazoline-6-sulfonate) radical scavenging activities and ferric reducing ability of low temperature-aged garlic (LTAG) were similar to those of raw garlic. LTAG also showed decreased lipopolysaccharide (LPS)-induced production of reactive oxygen species, although there were not significant differences among samples. In addition, xanthine oxidase activity was inhibited by LTAG; the 15 days and $60^{\circ}C$ extract showed outstanding inhibition compared with the others. To understand the molecular mechanisms behind the anti-oxidative activity of LTAG, we performed quantitative real-time PCR analysis. The 30 days and $70^{\circ}C$ extract upregulated mRNA expression of antioxidant enzymes such as Cu/Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase, and catalase in LPS-stimulated RAW 264.7 cells. This result indicates that LTAG can be a functional food as a nature antioxidant and antioxidant substance.

PROBLEMS IN OSTEOGENIC DIFFERENTIATION OF RAT BONE MARROW STROMAL CELLS (쥐의 골수로부터 추출한 줄기세포를 이용한 조골세포로의 분화 유도과정에서 나타난 문제점에 관한 분석 연구)

  • Kim, In-Sook;Cho, Tae-Hyung;Zhang, Yu-Lian;Lee, Kyu-Back;Park, Yong-Doo;Rho, In-Sub;Weber, F.;Lee, Jong-Ho;Kim, Myung-Jin;Hwang, Soon-Jung
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.1
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    • pp.1-8
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    • 2005
  • This study was aimed to characterize osteogenic potential of rat bone marrow stromal cells (BMSC) isolated with standard flushing method and investigate the plasticity of transdifferentiation between osteoblastic and adipocytic lineage of cultured BMSC. Unlike aspiration method in human, rat bone marrow was extracted by means of irrigation with culture media that elevates the possibility of co-extraction of committed osteoprogenitor, or preosteoblast or other progenitor cells of several types present inside bone marrow. The cultured stromal cells showed high ALP activity which is representative marker of osteoblast without any treatment. Osteogenic inducers such as Dex and BMP-2 were examined for the evaluation of their effect on osteogenic and adipocytic differentiation of stromal cells, because they function as osteoinductive agent in stromal cells, but simultaneously induce adipogenic differentiation. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity or mRNA expression of osteoblast markers such as osteopontin, bone sialoprotein, collagen type I and CbfaI, and in vitro matrix mineralization by von Kossa staining. Oil red staining method was used to detect adipocyte and adipocytic marker, aP2 and $PPAR{\gamma}2$ expression was examined using RT-PCR. It can be supposed that irrigation procedure resulted in high portion of already differentiation-committed osteoprogenitor cell showing elevated ALP activity and strong mineralization only under the supplement of $100{\mu}M$ ascorbic 2-phosphate and 10mM ${\beta}$-glycerophosphate without any treatment of osteogenic inducers such as Dex and BMP-2. Dex and BMP-2 seemed to transdifferentiate osteoprogenitor cells having high ALP activity into adipocytes temporarily, but continuous treatment redifferentiated into osteoblast and developed in vitro matrix mineralization. This property must be considered either in tissue engineering for bone regeneration, or in research of characterization of osteogenic differentiation, with rat BMSC isolated by the standard irrigation method.

Molecular Characterization of Metallothionein Gene of the Korean Bitterling Acheilognathus signifer (Cyprinidae) (묵납자루 (Acheilognathus signifer; Cyprinidae) metallothionein 유전자의 클로닝 및 특징 분석)

  • Lee, Sang-Yoon;Bang, In-Chul;Nam, Yoon-Kwon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.10-20
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    • 2011
  • Genetic determinant for metallothionein (MT), a cysteine-rich protein playing essential roles in metal detoxification and homeostasis, was characterized in the Korean bitterling (Acheilognathus signifer, Cyprinidae), an endemic fish species. The full-length A. signifer MT (AsMT) cDNA (551 bp) is composed of a single open-reading frame (ORF) to encode a polypeptide of 60 amino acids containing 20 cysteine residues whose positions are conserved in most cypriniform MTs. At the genomic level, the AsMT (2,593 bp spanning the 5'-flanking region to the 3'-untranslated region) represented a conserved tripartite (three exons interrupted by two introns) structure with AT-rich introns. The upstream regulatory region (-1,914 bp from the ATG initiation codon) of AsMT displayed various sites and motifs for transcription factors involved in the metal-mediated regulation and stress/immune responses. The AsMT transcript was ubiquitously detected in various organs with variable expression levels, where the ovary and intestine showed the highest expression, while the heart and skeletal muscle represented the lowest level. During an exposure to copper (immersion in $0.5\;{\mu}M$ Cu for 48 h), the levels of AsMT transcripts were significantly elevated in the liver (more than 3.5-fold), moderately in the gill, kidney, and spleen (ranging from 1.5- to 2.5-fold), and barely in the brain and intestine. Results of this study could form a useful basis to explore the metal-related stress physiology of this endangered fish species.

EXPRESSION OF TGF-${\beta}1$, OSTEONECTIN AND BMP-4 ON BONE REGENERATION IN MANDIBULAR DISTRACTION OSTEOGENESIS COMBINATION WITH A COMPRESSION STIMULATION (하악골 신장술에서 압축자극을 통한 골재생시 TGF-${\beta}1$, Osteonectin 및 BMP-4의 발현)

  • Park, Seong-Jin;Kim, Uk-Kyu;Chung, In-Kyo;Hwang, Dae-Seok;Kim, Yong-Deok;Shin, Sang-Hun;Kim, Cheol-Hoon;Byun, June-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.4
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    • pp.288-296
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    • 2007
  • Distraction osteogenesis(DO) is a technique of lengthning bone including soft tissue by gradual separation of surgically divided bone surfaces. Distraction osteogenesis combination with a compression stimulation(DO-CO) was a new technique by authors to enhance new bone quality and to shorten the consolidation period. The purpose of this study was to compare DO with DO combined with compression force in efficiency by evaluating the expression of TGF-${\beta}1$, osteonectin and BMP-4 on bone regenerate in rabbit mandible. Fourty two rabbits were used for this experiment. On the control group, the distraction was carried out at the rate of 1 mm per day to obtain the amount of 8 mm distraction for 8 days. On the experimental group, the distraction was carried out at the rate of 1 mm per day for 10 days, 3 days-latency period, and then the compression was carried out as counter direction 1 mm per day for 2 days. After 0 day, 5 days, 13 days, 20 days, 27 days, 34 days and 41 days, three rabbits on each group were sacrificed and the distracted portion of mandible were cut and treated for RT-PCR observation. The level of expression of TGF-${\beta}1$ and osteonectin were shown more and longer expression in the experimental group than in the control group. The expression of BMP-4 was maintained with high level during the entire experimental period in both groups. These findings suggested that DO with compression stimulation could be a favorable technique for obtaining a good new bone quality.