• Title/Summary/Keyword: clonality

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Characterizations of the Antimicrobial Resistant Determinants in Proteus spp. Isolated from Humans and Chickens in the Chungcheong Province (충청지역의 사람과 닭으로부터 분리된 Proteus속에 속하는 균주에 존재하는 항균제 내성유전자의 유전형 분석)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.4
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    • pp.327-334
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    • 2016
  • Recently, antimicrobial resistance of pathogenic bacteria has been increasing due to excessive use of antimicrobial agents in both humans and livestock. PCR amplification and nucleotide sequence analyses were conducted to investigate16S ribosomal RNA methyltransferase (RMTase) genes and integrons in P. mirabilis strains isolated from clinical specimens and chickens in an area of the Chungcheong providence. In addition, clonality analysis of P. mirabilis strains was performed using a repetitive extragenic palindromic sequence-based PCR (REP-PCR) method. Of the total 38 P. mirabilis isolates, 7 (18.4%) strains were isolated from clinical specimens contained in the RMTase genes and showed resistance to amikacin, tobramycin, and gentamicin. A total of 23 (60.5%) isolates carried class 1 integrons, but no isolates in our study harbored class 2 and class 3 integrons. Class 1 integrons detected in our study harbored genes encoding resistance to aminoglycosides (aadA2, aadA5, aadA7, and aacCA5), ${\beta}$-lactams ($bla_{PSE}$), erythromycin (ereA), lincosamides (linF), and trimethoprim (dfrA12, dfrA17, and dfrA32). We confirmed that the RMTase genes had spread among only the P. mirabilis isolates from clinical specimens, but class 1 integrons had widely disseminated among P. mirabilis isolates from clinical specimens and chickens. In addition, identical REP-PCR banding patterns were evidenced in only P. mirabilis isolates from chickens. Our results suggest the horizontal spreading of P. mirabilis isolates in the chicken farm. To prevent further spreading of antimicrobial resistant genes among P. mirabilis isolates, monitoring and clinical policing will be required.

Ex vivo Expansion and Clonal Maintenance of CD34+ Selected Cells from Cord Blood and Peripheral Blood (제대혈 및 말포혈로부터 분리한 CD34 양성 세포의 체외 증폭 및 클론 유지)

  • Kim, Soon Ki;Ghil, Hye Yoon;Song, Sun U.;Choi, Jong Weon;Park, Sang Kyu
    • Clinical and Experimental Pediatrics
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    • v.48 no.8
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    • pp.894-900
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    • 2005
  • Purpose : Because of the unavailability of marrow transplantation, umbilical cord blood (CB) is increasingly being used. We evaluated the potential of ex vivo expansion and clonality in CD34+ cells separated from cord blood source and mobilized peripheral blood (PB) in a serum-free media. Methods : The CD34+ cells, selected from CB and mobilized PB, were expanded with hematopoietic growth factors. They were then cultured for burst-forming units of erythrocytes (BFU-E), colony-forming units of granulocytes and monocytes (CFU-GM) and colony-forming units of megakaryocytes (CFU-Mk) at culture days 0, day 4, day 7, and day 14 with various growth factors. Results : The CB-selected CD34+ cells showed significantly higher total cell expansion than those from the PB at day 7 (2 fold increase than PB). The CB-selected CD34+ cells produced more BFU-E colonies than did the PB on culture at days 7 and at day 14. Also, the CB-selected CD34+ cells produced more CFU-Mk colonies than did the PB on culture at day 4 and at day 7. Conclusion : The ex vivo expansion of the CB cells may be promising in producing total cellular expansion, CFU-Mk and BFU-E compared with PB for 7 to 14 days. The growth factors combination including megakaryocyte growth and development, flt3-ligand and interleukin-3 showed more expansion in the view of total cells and clonal maintenance compared with less combination.

$pep^{27}$ and lytA in Vancomycin-Tolerant Pneumococci

  • Olivares, Alma;Trejo, Jose Olivares;Arellano-Galindo, Jose;Zuniga, Gerardo;Escalona, Gerardo;Vigueras, Juan Carlos;Marin, Paula;Xicohtencatl, Juan;Valencia, Pedro;Velazquez-Guadarrama, Norma
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1345-1351
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    • 2011
  • Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and $pep^{27}$ are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and $pep^{27}$ were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in $pep^{27}$ expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, $pep^{27}$ expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in $pep^{27}$ expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.

A Case of Primary Pulmonary Low-grade B-cell Lymphoma of Bronchus-associated lymphoid tissue with Bilateral Consolidation and Indolent Clinical Course (양측성 고형질화 음영을 보인 무증상의 기관지-관련 림프양 조직의 저등급 B-세포 원발성 폐 림프종 1례)

  • Kim, Yang-Ki;Kim, Chul;Jean, Jin;Jean, Ki-Won;Kim, Dong-Won;Lee, Dong-Wha;Hong, Dae-Sik;Park, Hee-Sook;Uh, Soo-Taek;Kim, Yong-Hoon;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.5
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    • pp.1073-1081
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    • 1998
  • Primary pulmonary lymphoma (PPL) is an uncommon tumor, which constitutes 3-4% of all extranodal lymphomas and 0.3-0.5 % of all primary pulmonary malignant tumors. Low-grade B-cell lymphomas of bronchus-associated lymphoid tissue (BALT) accounted for the majority of PPL. This BALT lymphomas are frequently asymptomatic and have an excellent prognosis and an indolent clinical course by contrast with T-cell type. Therefore, determination of the B- or T-immunophenotype of the tumor cells is known to be very important from a clinical aspect. Recent advances in immunohistochemical techniques, cytogenetics, and molecular biology have allowed better definition of type, maturation, and clonality of lymphoma cells and have made it possible to better understand the PPL. We experienced an asymptomatic 43-year-old man who was evaluated for infiltrates on both sides discovered incidentallly after a routine chest roentgenogram. He was eventually diagnosed as low-grade B-cell lymphoma of BALT by immunohistochemical staining from specimens obtained by open lung biopsy. He was treated with combination chemotherapy. At follow up 12 mons following initial diagnosis, he remains in stable. We report this case, who showed a relatively favorable prognosis and indolent clinical course compatible with low-grade B-cell lymphoma.

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Molecular Typing of Salmonella enterica serovar Typhi Strains Isolated in Busan by Pulsed-Field Gel Electrophoresis (부산지역에서 분리된 Salmonella enterica serovar Typhi균에 대한 PFGE를 이용한 Molecular typing)

  • Min, Sang-Kee;Lee, Ju-Hyun;Park, Eun-Hee;Kim, Jung-Ah;Kim, Kyu-Won
    • Journal of Life Science
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    • v.16 no.4
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    • pp.664-671
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    • 2006
  • We obtained 424 Salmonella enterica serovar Typhi isolates from sporadic cases of infection in Busan during 1996 to 2005. We investigated the trend of antimicrobial resistance and molecular typing by pulsed-field gel electrophoresis (PFGE). Of the total 424 isolates, 6 strains (1.4%) were multidrug-resistant (MDR) S. enterica serovar Typhi isolates, 2 strains (0.5%) were resistant to only nalidixic acid, and the remaining 416 strains (98.1%) were fully susceptible to the 18 antimicrobial agent. PFGE of XbaI-digested chromosomal DNA was performed on 50 sporadic S. enterica serovar Typhi isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that these isolates were much more heterogeneous and at least 32 different PFGE patterns were generated according by dice coefficient, between 0.69 and 1.0. Restriction fragment patterns consisted of 13 to 18 fragments ranged in size from 20 to 630 kb. The results confirmed that PFGE would be an useful tool for investigating surveillance of sporadic or outbreak case and assessing clonality for S. enterica serovar Typhi in Busan area. Our finding will be valuable in developing rational strategies to control this pathogen and setting the basis of an effective PulseNet system in Korea.

Selective Expansion of TCR $V{\beta}3$+CD4+T Cells in Collagen-induced Arthritis in DBA/1 Mice (콜라겐 유도 관절염에서 콜라겐 항원 특이 $V{\beta}3$+CD4+T 세포의 선택적 증식)

  • Lee, Jae-Seon;Cho, Mi-La;Lee, Jung-Eun;Min, So-Youn;Yoon, Chong-Hyeon;Kim, Wan-Uk;Min, Jun-Ki;Park, Sung-Hwan;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.78-88
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    • 2005
  • Background: Collagen-induced arthritis (CIA) in mice is animal model of autoimmune disease known as rheumatic arthritis in human. We investigated CII-specific CD4+ T cell receptor usage in CIA mice. Methods: In CIA model, draining lymph node (dLN) CD4+ T cells and splenocytes at $3^{rd},\;5^{th},\;8^{th}$ week, we investigated CII-specific T cell proliferation, production of IL-17, IFN-${\gamma}$, TNF-${\alpha}$, IL-4 and IL-10. And we also performed anti-CII IgG Ab measurements in serum level, TCRV ${\beta}$ usage and T cell clonality with RT-PCR-SSCP analysis. Also, we performed proliferative response against CII when CII-specific T cell subset is deleted. Results: CIA mice showed more increase in the serum level of anti-CII IgG than normal mice after induction of arthritis. And the level of anti-CII IgG2a in CIA mice was increased after $3^{rd}$ week after primary immunization, while anti-CII IgG1 was decreased. Draining LN CD4+ T cells have proliferated against CII stimulation at $3^{rd}$ week after $1^{st}$immunization. CD4+T cells derived from dLN of CIA mice produced proinflammatory cytokine IFN-${\gamma}$, IL-17 etc. Draining LN CD4 T cells of CIA presented higher proportion of CD4+V ${\beta}3$+subset compared to those of normal mice at $3^{rd}$ week after $1^{st}$ immunization, and they were increased in proportion by CII stimulation. Draining LN CD4+ T cells without TCRV ${\beta}3+/V{\beta}8.1/8.2+/V{\beta}$10b+cells were not responsive against CII stimulation. But, CII-reactive response of TCRV ${\beta}3-/V{\beta}8.1/8.2-/V{\beta}$10b- T cells was recovered when $V{\beta}3+$ T cells were added in culture. Conclusion: Our results indicate that CD4+$V{\beta}3+$ T cells are selectively expanded in dLN of CIA mice, and their recovery upon CII re-stimulation in vitro, as well as the production Th1-type cytokines, may play pivotal role in CIA pathogenesis.

Genetic Monomorphism of the Natural Ilex cornuta Community at the Northern Range Limit in Buan, Jeollanam-do in Korea Revealed by AFLP Markers (AFLP 표지자에서 나타난 전라남도 부안의 북방한계지에 자생하는 호랑가시나무 군락의 유전적 단형성)

  • Hong, Kyung Nak;Park, Yu Jin;Lee, Jei Wan;Kim, Young Mi
    • Journal of Korean Society of Forest Science
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    • v.104 no.2
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    • pp.187-192
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    • 2015
  • Individuals in the population under a particular environmental condition influencing recurrently for a long time could locally adapted and local adaptation is of a fundamental importance in a breeding program, conservation activities of genetic resources or evolutionary biology. Plants at northern range limits have higher probability of expressing an adaptative genetic trait. The natural community at the northern range limit of Ilex cornuta (Chinese holly) in Buan, Jeollanam-do in Korea was composed of adults of 744 and seedlings of 211 (hereafter Community) and is designated as the Korean Natural Monument (No. 122) by the law. At two adjacent areas to Community, 85 (hereafter Plantation I) and 27 hollies (hereafter Plantation II) were planted respectively for preparations of the next generation. Eighty-five trees were sampled for genetic analysis in the three groups. Fifty-two (36%) of the total 143 amplicons were polymorphic from four AFLP primer combinations. A total of thirteen genotypes was identified and just one genotype was for 52 trees of Community. Seven and five genotypes were shown for Plantation I and II, respectively. There was no identical genotype between Community and Plantation (I or II) or between two plantation groups. Number of private loci was 2 for Community, 6 for Plantation I and 4 for Plantation II. We presumed their genetic backgrounds were quite different with one another and the plantation groups were made independently because they were different not only the genetic compositions but also their ages. Considering the genetic monomorphism by AFLP markers, observations of only male trees and asexual propagation as layerage or cuttage, the hollies in Community might be a genet by root suckering from a single male tree, not the results of selective removal of female trees for ornamental use in the past.