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Effects of SIS Sponge and Bone Marrow-Derived Stem Cells on the Osteogenic Differentiation for Tissue Engineered Bone (SIS 스폰지와 골수유래줄기세포를 이용한 조직공학적 골분화 유도)

  • Park Ki Suk;Jin Chae Moon;Yun Sun Jung;Hong Keum Duck;Kim Soon Hee;Kim Moon Suk;Rhee John M.;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.29 no.5
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    • pp.501-507
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    • 2005
  • Small intestinal submucosa (SIS) had been widely used as a biomaterial without immune rejection responses. SIS sponges prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). SIS powders dissolved in $3\%(v/v)$ acetic acid aqueous solution for 48hrs and freeze-dried. EDC solution ($H_2O$ : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100mM. In vitro, rat-BMSCs seeded in SIS sponges and induced the osteogenesis for 28 days. We have characterized the osteogenic potential of rat-BMSCs in SIS sponges by alkaline phosphatase activity(ALP), n assay, SEM and RT-PCR for osteogenic phenotype. In SEM, all morphology of SIS sponges was regular and showed interconnected pore structure. By RT-PCR analysis, we observed type I collagen expression. These results demonstrate osteogenic differentiation of rat-BMSCs. In conclusion, we confirmed that the morphology of surface, cross-section, and side of SIS sponges were highly porous with good interconnections between each pores, which can support the surface of cell growth, proliferation, and differentiation. This result indicates that SIS sponge is useful for osteogenesis of BMSCs.

Characteristic, Genesis and Classification of Soils Derived from Coarse Grain Granitic Materials (조립질(粗粒質) 화강암(花崗巖) 토양(土壤)의 특성(特性)과 생성(生成)·분류(分類))

  • Jung, Sug-Jae;Hyeon, Geun-Soo;Moon, Yong-Taik;Jo, Young-Kil
    • Korean Journal of Soil Science and Fertilizer
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    • v.27 no.1
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    • pp.3-9
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    • 1994
  • Characterstics, genesis and classification of soils derived from coarse grain granitic materials were discussed with four soil series, such as Samgag, Sangju. Sachon and Yecheon which were distributed over the area of Gangdae-Ri, Nengseo-Myeon, Yeoju-Gun, Gyunggi-Do. The results are as follows. 1. Samgag, Sangju, Sachon and Yecheon had a soil of excessively, well, imperfectly and poorly drained, thus they had a soil drainage sequence. 2. Soil textural class were from sandy loam to loam. Silt and clay content were increased with descending to the local bottom, while sand content was decreased. 3. Soils were very strongly to strongly acid and OM, CEC, exchangeable cation, and available $P_2O_5$ in soils seemed to be increased with ascending to the local boctom. 4. Kaolinite and Quartz were the dominant clay mineral and the other was Vermiculite and Illite. 5. Samgag was classified as Typic Dystrochrepts, Sangju as Dystric-Fluventic Eutrochrepts, Sachon as Aeric-Fluventic Halpaquepts, and Yecheon as Fluventic Haplaquepts.

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Differential Gene Expression in the Bovine Transgenic Nuclear Trasnsfer Embryos (소 형질전환 복제란의 유전자 이상발현 규명)

  • Cho, Jong-Ki;Song, Bong-Seok;Yong, Hwan-Yul;Lee, Doo-Soo;Koo, Deok-Bon;Lee, Kyung-Kwang;Shin, Sang-Tae
    • Journal of Veterinary Clinics
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    • v.24 no.3
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    • pp.295-299
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    • 2007
  • The detrimental effects of gene transfection on embryo development and the molecular mechanism behind the differential expression of genes related to early embryo development were assessed in the production of transgenic cow embryos through somatic cell nuclear transfer (NT). Parthenogenetic, IVF, and transgenic NT embryos derived from ${\alpha}_1$-antitrypsin transfected ear fibroblast cells was produced. To investigate the molecular mechanism behind lower developmental competence of transgenic NT embryos, the differential mRNA expression of three genes ($IFN-{\tau}$, Oct4, Fgf4) in the 3 types of embryo (Parthenogenetic, IVF, transgenic NT) was examined. RNA was extracted from ten blastocysts derived from 3 types of embryos and reverse-transcripted for synthesis of the first cDNA. The quantification of 3 gene transcripts ($IFN-{\tau}$, Oct4, and Fgf4) was carried out in three replicate by quantitative real-time reverse transcriptase PCR. Expression level of $IFN-{\tau}$ mRNA was significantly higher in transgenic NT embryos than parthenogenetic and IVF embryos (P<0.05). However, expression level of Oct4 and Fgf4 of transgenic NT embryos was significantly lower than IVF embryos (P<0.05). Altered levels of these three mRNA transcripts may explain some of the embryonic/fetal/neonatal abnormalities observed in offspring from transgenic NT embryos.

Surveillance of viable Acanthamoeba spp. and Naegleria fowleri in major water sources for tap water in Korea (한국 주요 상수원수에서의 가시아메바와 파울러자유아메바 조사)

  • Kim, Min-jeong;Lee, Gyu-Cheol;Kim, Kunwoo;Lee, Hyunji;Kim, Min Young;Seo, Dae Keun;Lee, Jeong Yeob;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.237-243
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    • 2018
  • The pathogenic free-living amoebas (FLAs), Acanthamoeba spp. and Naegleria fowleri, can cause fatal infections, including amoebic encephalitis. They are ubiquitously distributed in nature, including in diverse bodies of water. In order to survey Acanthamoeba spp. and N. fowleri in source water in Korea, we used culture-based real-time PCR to detect viable FLAs in 52 source water samples collected between July 2017 and December 2017. Acanthamoeba spp. and N. fowleri were detected in 42 samples (80.8%) and 6 samples (11.5%), respectively. Acanthamoeba spp. were detected at approximately the same frequency in all seasons, but N. fowleri was mainly detected in summer and autumn, with no N. fowleri detected in winter. These results demonstrate that these pathogenic FLAs, especially N. fowleri, which has caused deaths in the United States and China, are widely distributed in the Korean aquatic environment.

Clinical and Laboratory Finding of the 2009 Pandemic influenza A (H1N1) in Children (소아에서 2009 신종 인플루엔자 A (H1N1) 바이러스 감염의 임상적 특징)

  • Sohn, Yu Rak;Park, Su Hyun;Kim, Won Duck
    • Pediatric Infection and Vaccine
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    • v.18 no.2
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    • pp.173-181
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    • 2011
  • Purpose : 2009 Pandemic influenza A (H1N1) virus was identified in March 2009 and subsequently caused worldwide outbreaks. We described the clinical and epidemiological characteristics of H1N1 influenza infection. Methods : We used retrospective medical chart reviews to collect data on the visiting patients from a single institute. H1N1 infection was confirmed in specimens with the use of a RT-PCR (real time reverse transcriptase polymerase chain reaction assay). Result : 6,836 patients had H1N1 RT-PCR test, and 2,781 were confirmed with H1N1 virus infection. 158 patients (5.7%) had hospital treatment and inpatients were significantly younger (5.4${\pm}$3.3 years) than outpatients (7.5${\pm}$3.9 years) among H1N1 virus confirmed patients. Oxygen, steroid, immunoglobulin, ventilator treatment was provided in a substantial proportion among pneumonia patients accompanying wheezy respiration. In addition more intensive care was needed in patients accompanying segmental, lobar, interstitial, mixed pneumonia and lung effusion (27.2%) than patients with bronchopneumonia (7.3%) among H1N1 virus infection confirmed patients. Seventy-one infants had oseltamivir treatment out of 83 infants under 1 year, and no significant side effects and complications were identified. Conclusion : In 2009 pandemic influenza A (H1N1), hospital treatment was needed in younger patients. Early intensive care was needed in pneumonia patients accompanying wheezy respiration, and patients accompanying segmental, lobar, interstitial, mixed pneumonia and lung effusion.

Epidemiology and Clinical Severity of the Hospitalized Children with Viral Croup (바이러스성 크루프로 입원하는 소아 환자의 역학적 특성과 임상적 중증도 평가)

  • Jeon, In Soo;Cho, Won Je;Lee, Jeongmin;Kim, Hwang Min
    • Pediatric Infection and Vaccine
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    • v.25 no.1
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    • pp.8-16
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    • 2018
  • Purpose: In this study, the clinical and epidemiological characteristics of patients admitted for viral croup were analyzed to evaluate disease severity based on the organism that caused the infection. Methods: We retrospectively reviewed the medical records of 302 patients who were admitted to the Department of Pediatrics at the Wonju Severance Hospital between May 2013 and December 2016 for viral croup. Patients who showed positive results on multiplex polymerase chain reaction were subsequently diagnosed with respiratory virus infection. The Westley scoring system was used to evaluate the severity of viral croup. Results: Of the 302 patients, 149 were admitted due to severe viral croup, including 88 boys and 61 girls, with a boy-to-girl ratio of 1.44:1. About 110 cases of parainfluenza virus infection have been reported, which accounted for almost half of the total cases. The other identified viruses included influenza virus, human rhinovirus, and respiratory syncytial virus. Analysis of the association between severe viral croup and causative pathogen revealed that only parainfluenza type 2 virus showed a significantly high risk. Parainfluenza type 2 virus did not show an age-based difference in frequency but showed relatively a higher frequency of infections during the summer and fall. Conclusions: In this study, parainfluenza virus type 2 was the only virus associated with severe viral croup. To facilitate proper preventive management, treatment, and prognosis evaluation of viral croup, prospective and multicenter studies should assess the additional variables and the severity of the virus. Additionally, further studies should be conducted to assess age-dependent influences, as well as the regional and seasonal incidence of viral infection.

Molecular Serotyping of Group B Streptococcus Isolated from the Pregnant Women by Polymerase Chain Reaction and Sequence Analysis (임신부에서 분리된 B군 연구균의 중합효소연쇄반응과 염기서열분석을 통한 혈청형 분석)

  • Oh, Chi Eun;Jang, Hyun Oh;Kim, Nam Hee;Lee, Jina;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.16 no.1
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    • pp.47-53
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    • 2009
  • Purpose : This study was performed to investigate the serotype distribution of group B streptococcus (GBS) isolated from pregnant Korean women using molecular methods. Methods : The study materials included 42 GBS isolates obtained from the vagina and anorectum of pregnant women in Goyang, Korea between 2005 and 2006. Four clinical isolates with known serotypes (Ia, Ib, III, and V) were used for validation of molecular serotyping. We used serotype-specific primers for identification of the serotypes (Ia, Ib, III, V, and VI). To determine the ambiguous serotypes by serotype-specific PCR, sequence analysis of the PCR amplicons which had been amplified with GBS-common primers was used. Results : The serotypes determined by the molecular methods agreed with the previously known 4 serotypes (Ia, Ib, III, and V). The serotypes of all 42 isolates were successfully determined by molecular methods. The distribution of the GBS serotype was as follows in order of frequency: serotype III was found in 12 isolates (28.6%), serotype V was found in 11 isolates (26.2%), serotype Ia was found in 11 isolates (26.2%), serotype VI was found in 4 isolates (9.5%), serotype Ib was found in 2 isolates (4.8%), and serotype II was found in 2 isolates (4.8%). Conclusion : Serotypes III, V, and Ia were the most frequently identified serotypes in pregnant Korean women. Molecular serotyping is useful for surveillance of the serotype distribution of GBS in colonized pregnant women and GBS diseases of neonates.

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Phylogenetic Groups and Virulence Factors of Escherichia coli Causing Urinary Tract Infection in Children (소아 요로감염의 원인 Escherichia coli 균의 계통 분류와 독성인자 분석)

  • Kim, Ji Mok;Cho, Eun Young;Lee, Jae Ho
    • Pediatric Infection and Vaccine
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    • v.22 no.3
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    • pp.194-200
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    • 2015
  • Purpose: Urinary tract infection (UTI) is a common bacterial infection in children and Escherichia coli is a predominant pathogen. The purpose of this study is to evaluate phylogenetic groups and virulence factors of E. coli causing UTI in children in Korea. Methods: From October 2010 to April 2013, urinary E. coli strains were isolated from the 33 pediatric patients of UTI. Multiplex polymerase chain reactions were performed to evaluate the phylogenetic groups and 5 virulence factor genes (fimH, sfa, papA, hylA, and cnf1) of E. coli. Distribution of molecular characteristics of E. coli was analyzed by clinical diagnosis and accompanying vesicoureteral reflux (VUR). Results: Most (84.8%) uropathogenic E. coli were belonged to phylogenetics group B2 and the others (15.2%) were belonged to group D. The virulence factors were distributed as: fimH (100%), sfa (100%), hylA (63.6%), cnfI (63.6%), and papA (36.4%). According to clinical diagnosis, phylogenetic distribution of E. coli strain was 92.3% of B2 and 7.7% of D in acute pyelonephritis and 57.1% of B2 and 42.9% of D in cystitis. Distribution of virulence factors was similar in both groups. In patients with acute pyelonephritis, phylogenetic distribution was similar in VUR and non-VUR group, but proportion of papA genes were lower in VUR group than that of non-VUR group (43.8% vs. 20.0%, P=0.399). Conclusions: This study provides current epidemiologic molecular data of E. coli causing pediatric UTI in Korea and will be a fundamental for understanding the pathogenesis of pediatric UTI.

Molecular Sex Determination Using Sexual Dimorphisms between ZFX and ZFY Genes in Korean Hares(Lepus coreanus Thomas) (한국멧토끼 ZFX와 ZFY 유전자의 성별 이형성과 분자 성판별)

  • Han, Sang-Hyun;Cho, In-Cheol;Lee, Sung-Soo;Oh, Moon-You;Oh, Hong-Shik
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.402-406
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    • 2007
  • This study was performed to develop the molecular marker for sex determination of hare (Lepus coreanus) distributed in Korea which focused on sexual dimorphism between X and Y chromosomal homologous genes, zinc finger-X (ZFX) and -Y (ZFY). The intron 7 regions of ZFX and ZFY genes exhibited differential amplification patterns between male and female hares. The lengths of intron 7 region of ZFX and ZFY genes were 538 and 233-bp, respectively. Especially, the ZFX intron 7 contained a repetitive sequence identified as member of RNA-mediated transposable elements which was similar to CSINE2 commonly found in the rabbit genome. However, it was not present in intron 7 of ZFY gene. The molecular sex typing by polymerase chain reaction (PCR) was also carried out to determine the sex of hare based on difference in lengths between the intron 7 regions of ZFX and ZFY genes. All DNA samples tested had common band amplified from ZFX. However, the male hare DNAs had two distinct bands which amplified from ZFX and ZFY genes, respectively. The results from ZFX-ZFY PCR sex typing were identical to those from phenotypic investigation and from amplification patterns using male-specific sex determining region Y (SRY) gene as well. Finally, this study suggested that the sexual dimorphism between intron 7 regions of ZFX and ZFY could be useful genetic marker to determine sex of hare.

Enzymatic Characterization of a Thermostable 4-α-Glucanotransferase from Thermotoga neapolitana (Thermotoga neapolitana 유래 내열성 4-알파-글루칸전이효소의 효소적 특성)

  • Choi, Kyoung-Hwa;Seo, Ja-Yeong;Kim, Ji-Eun;Cha, Jae-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.221-226
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    • 2011
  • The gene encoding 4-$\alpha$-glucanotransferase (mgtA) from Thermotoga neapolitana was cloned and expressed in Escherichia coli in order to investigate whether this enzyme was capable of producing cycloamylose for industrial applications. MgtA was purified to homogeneity by HiTrap Q HP and Sephacryl S-200 HR column chromatographies. The size of the enzyme as determined by SDS-PAGE was about 52 kDa, which was in good agreement with its deduced molecular mass of 51.9 kDa. The optimal temperature and pH for the activity of the 4-$\alpha$-glucanotransferase was found to be $85^{\circ}C$ and 6.5, respectively. The enzyme hydrolyzed the 1,4-$\alpha$-glucosidic bonds in oligomeric 1,4-$\alpha$-glucans and transferred oligosaccharides (maltotriose being the shortest one) to acceptor maltodextrins. However, the enzymes had no activity against pullulan, glycogen, and other di- or trioligosaccharides with rare types of $\alpha$-bond. MgtA is distinguished from 4-$\alpha$-glucanotransferase from Thermotoga maritima in that it can convert maltotriose into maltooligosaccharides. The treatment of glucoamylase after the reaction of MgtA with maltotriose, maltotetraose, maltopentaose, or maltohexaose as sole substrate revealed that MgtA yielded linear maltooligosaccharides instead of cycloamylose.