• 제목/요약/키워드: Whole blood culture

검색결과 32건 처리시간 0.025초

인체 전혈 모델을 이용한 세포내 결핵균 살균력에 관한 연구 (Measuring Intracellular Mycobacterial Killing Using a Human Whole Blood Assay)

  • 천선희;송호연;이은희;오희정;강인숙;조지윤;홍영선
    • Tuberculosis and Respiratory Diseases
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    • 제53권5호
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    • pp.497-509
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    • 2002
  • 연구배경 : 대표적 세포내 감염질환인 결핵에 대한 숙주의 방어기전 및 면역반응은 아직도 정확히 이해되지 못하고 있으며 이러한 병리기전을 연구하기 위하여서는 적절한 감염모델이 필요하다. 전혈 (whole blood)은 체액성 면역과 세포성 면역을 모두 포함한 생체의 상태를 반영하므로 다양한 대상에서 면역상태의 차이에 따른 개체간 결핵균 살균력의 차이를 비교 할 수 있는 적절한 모델로 추정된다. 따라서 본 연구의 목적은 인체의 결핵균 전혈 배양모델을 개발하여 궁극적으로 시험관내에서 숙주면역의 정도를 측정하는 대리 표지자를 개발하고자하는 것이다. 방 법 : PPD 양성 정상인을 대상으로 제대혈, 결핵환자, 당뇨 및 폐암환자와 비교하였다. 전혈을 희석하여 결핵균 Mycobacterium avium과 M. tuberculosis $H_{37}Ra$에 낮은 감염률로 감염시키고 $37^{\circ}C$, 5% $CO_2$ 배양기 속의 회전교반기에서 회전시키면서 배양(rotating culture) 하였다. 배양 1 일, 3일 및 4일 뒤 증류수로 긴장저하용해 시킨 후 Middlebrook 7H10/OADC 평판배지에서 결핵균 집락이 형성될 때까지 3-4주간 $37^{\circ}C$, 5% $CO_2$ 배양기에 배양하여 집락수를 계산하였다. 일부실험에서 TNF-${\alpha}$의 분비능을 90%이상 감소시키 기 위하여 methylpredmsolone과 pentoxifylline을 첨가하여 면역조정을 하였으며, CD4+ T-림프구와 CD8+ T-림프구를 magnetic bead에 코팅된 단클론 항체를 사용하여 제거하였다. 결핵균의 수는 용해질 $m{\ell}$ 당 CFU로 계산하였다. 살균력은 ${\Delta}$ log killing ratio로 표시하였다. ${\Delta}$ logKR=$log_{10}$(Final CFU/Initial CFU). 결 과 : 1. 제대혈의 결핵균 살균력이 PPD 양성 대조군에 비하여 다소 감소된 경향을 보였으며, 결핵환자의 결핵균 살균력은 PPD 양성 대조군과 특별한 차이를 보이지 않았다. 또한 당뇨군과 폐암군의 결핵균 살균력도 정상 대조군에 비하여 특별한 차이를 보이지 않았다. 2. Methylprednisolone과 pentoxyfylline을 사용한 면역조정 시에 제대혈, PPD 양성 정상 대조군과 결핵군 모두에서 전혈에서의 결핵균의 살균력이 감소되는 경향을 보였다. 3. CD4+ 및 CD8+ T-림프구 삭제시 log KR가 증가되어 유의하게 결핵균 살균력이 감소되었으며 CD4+ 및 CD8+ T-림프구 동시 삭제시 현저한 상승효과를 보였고 이러한 결과는 Mycobacterium avium보다는 독력이 없는 균인 Mycobacterium tuberculosis $H_{37}Ra$에서 보다 뚜렷하였다. 4. 폐결핵 치료후의 결핵균 살균력은 치료 전과 비교하여 유의하게 ${\Delta}$ logKR가 감소하였으며, 배양 3-4일에도 현저한 결핵균 증식의 억제를 보였다. 결 론 : 인체의 결핵균에 대한 감수성인 개체간의 면역상태를 전혈에서 결핵균 살균력을 통하여 비교할 수는 없었다. 그러나 인체 전혈 모델은 간단하고 임상경과 관찰이 쉬우며 결핵환자에서 치료 전과 후에 현저한 결핵균 살균력의 차이를 보이므로, 최근 결핵연구의 가장 중요한 과제의 하나인 백신 개발에서 그 성과를 판단하는 vaccine trial에 이용할 수 있을 가능성을 시사한다.

Mercury chloride 및 Methylmercury chloride가 정상인(正常人)의 혈액배양(血液培養)에서 임파구(淋巴球)의 자매염색분체교환(姉妹染色分體交換)에 미치는 영향(影響) (Sister Chromatid Exchanges in Lymphocytes on Normal Human Blood Culture with Mercury chloride or Methylmercury Chloride)

  • 고대하
    • Journal of Preventive Medicine and Public Health
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    • 제17권1호
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    • pp.245-250
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    • 1984
  • Reciprocal exchanges of DNA in sister chromatids (SCEs) are induced by various carcinogens and mutagens, although the quantitative relationship between the number of mutations and SCEs induced varies among chemicals. Nevertheless, the analysis of SCEs production by various agents often proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity. Mercury, even if which has no evidences for mutagenicity and carcinogenicity, is reported to exert some cytotoxic effects, such as chromosomal aberrations or bad influences to ovulation and reproduction in experimental animals, etc.. In this study, tests for sister chromatid exchanges have been carried out on normal human lymphocytes in whole blood culture to add mercury chloride ($HgCl_2$) or methylmercury chloride ($CH_3\;HgCl$) for 72 hr. The results indicate the dose-dependent relationship between the frequencies of SCEs and the concentrations of $HgCl_2,\;CH_{3}HgCl$ and 5-bromo-2'-deoxyuridine (BrdU). Lymphocyte proliferation has depressed in the higher concentration of mercury.

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Phytomitogen에 의한 토끼 임파구의 blast transformation I. 유사분열에 미치는 배지, 유사분열촉진물질 및 배양시간의 효과 (Phytomitogen induced blast transformation of rabbit I. Effect of medium, phytomitogen and culture hours on the uptake of 3H-thymidine)

  • 김종수;김충희
    • 대한수의학회지
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    • 제31권1호
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    • pp.49-53
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    • 1991
  • The present study has been carried out to investigate the optimal condition on lymphocyte blastogenesis of rabbit lymphocytes, whole blood culture and microculture system in conjunction with a semiautomatic multiple sample harvester(SAMSH) was used to study the In vitro optimal condition of rabbit lymphocytes. Data were presented to show many variables that are involved in studying the phytohemagglutinin(PHA) and lipopolysaccharide(LPS) response of rabbit lymphocyte in a microculture system. Analysis indicated that the conditions for optimal PHA as measured by incorporation of $^3H$-TdR include: (1) use of RPMI-1640 as culture medium. (2) use of $6{\mu}g$ of PHA, per culture. (3) 48-hours culture period. Conditions for optimal stimulation with LPS mitogen were similar to those used for PHA.

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초국가적 입양과 탈경계적 정체성 -제인 정 트렌카의 『피의 언어』 (Transnational Adoption and Beyond-Borders Identity: Jane Jeong Trenka's The Language of Blood)

  • 김현숙
    • 영어영문학
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    • 제57권1호
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    • pp.147-170
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    • 2011
  • This paper elucidates the characteristics of transnational adoption, estimates the possibility of beyond-borders identity of transnational adoptees, and tries to analyze Jane Jeong Trenka's The Language of Blood in its context. Though it has been regarded as one of the most humanitarian ways of helping orphans and poor children of the world, transnational adoption, a one-way flow of children from poor Asian countries to rich white countries, has been operated under the market logic between countries. Transnational adoptees, who had been abandoned and forced to be taken away from their birth mother, and later, to fulfill the desire of white parents for a perfect family, perform an ideological labor, serving to make the heterogeneous nuclear family complete. Korean transnational adoptees, forced to transcend the borders of nation, culture, and ethnicity, experience racial conflict and alienation in white adoptive family and society. Their diaspora experience of violent dislocation creates frustration and confusion in establishing their identity as a whole being. When they return to Korea to find their birth mother and their true identity, Korean adoptees, however, are faced with other obstructing issues, such as language problem, culture conflict, and maternal nationalism. Finally, Korean transnational adoptees reject Korean nationalism discourse based on blood, and try to redefine themselves as beyond-borders subjectivities with new and fluid identities. Jane Jeong Trenka's The Language of Blood, an autobiographical novel based on her experiences as a transnational adoptee, represents a Korean adopted girl's personal, cultural, and racial conflict within her white adoptive family, and questions the image of benevolent white mother and the myth of multiculturalism. The novel further represents Jane's return to Korea to find out her true identity, and shows Jane's disappointment and alienation in her birth country due to her ignorance of language and culture. Returning to USA again, and trying to be reconciled with her American mother, Jane shows the promise of accepting her new identity capable of transcending the borders, and thus, the possibility of enlarging the category of belonging.

개 혈액 재료에서의 Brucella 검출을 위한 진단방법의 비교 (Comparison of diagnostic methods for detection of Brucella species in dog blood samples)

  • 권순오;람쯩꽝;허문;안동춘;박상희;박미연;이영주;한태욱
    • 한국동물위생학회지
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    • 제32권4호
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    • pp.335-341
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    • 2009
  • Canine brucellosis produce abortions and infertility in dogs and is currently diagnosed by serological methods such as rapid slide agglutination test with 2-mercaptoethanol (2-ME RSAT) and immunochromatographic assay (ICA). Bacterial isolation is considered gold standard for Brucella diagnosis and the polymerase chain reaction (PCR) is an alternative method to bacterial isolation. A total of 36 whole blood samples were collected from dogs reared in area of Chuncheon and were subjected to serology (2-ME RSAT and ICA for B. canis, Rose Bengal test and C-ELISA for B. abortus), blood culture and 3 types of PCRs (BSCP31, 16s rRNA, and OMP-2). All blood samples were negative by serology and blood cultures. The BCSP31 and the OMP-2 PCR detected 5 samples were positive whereas the 16S rRNA PCR detected all samples were negative as serological methods and blood culture did. From the results observed in the present study, we conclude that 16S rRNA PCR could be used for direct PCR for canine blood samples.

Effect of Mitogens, Supplement of β-mercaptoethanol and Fetal Bovine Serum Supplementation in Whole Blood Culture Medium for Bovine Chromosome Analysis

  • Takagi, M.;Tsuno, M.;Ohtani, M.;Acosta, T.J.;Miyamoto, A.;Miyazawa, K.;Sato, K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권5호
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    • pp.615-621
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    • 2002
  • We investigated the effects of the mitogen supplements of 3 types, pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (ConA), to a whole blood culture system on the number of metaphase spreads obtained in perinatal bovine chromosome analysis. In addition, the supplementation of ${\beta}$-mercaptoethanol (${\beta}$-ME) and FBS was examined in such system. Significant differences (p<0.05) were seen in the number of metaphase spreads with PHA stimulation compared to both PWM and ConA stimulation. When examined the effects of ${\beta}$-ME supplementation, the number of metaphase spreads was significantly (p<0.05) increased at $30{\mu}M$ ${\beta}$-ME compared to control. When evaluated FBS supplementation during PWM stimulation, no significant effect of the supplementation was found. Finally, the effects of the cortisol concentration (10-20, 20-30 and >30 ng/ml) of the blood samples were examined. There was no significant effect of cortisol concentration (p>0.05) among these 3 cortisol concentration groups. The mean percentages of normal metaphase plates (2n=60) from each calf 1) with ${\beta}$-ME, 2) without ${\beta}$-ME and 3) with FBS stimulated with PWM were not significantly different (p>0.05). In conclusion, these findings may be useful in cytogenetic screening programs for not only perinatal calves but also for mature cattle.

생명 문화 정립을 위한 시론적 모색 (Contemporary Explorations to Establish Life Culture)

  • 이재복
    • 비교문화연구
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    • 제21권
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    • pp.165-188
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    • 2010
  • One of the things that should be done first in establishing the cosmic life culture is to reflect on the old human-oriented culture. If the cosmic life culture absolutizes everything except for humans according to the logic of reason, its foundation will weaken or eventually get lost. Separating humans from the universe is just like separating life from it. Given that all life, whether it is humans or the earth, originated from the universe, such an effort for separation merely values an individual life by excluding all life or whole life. When the human body and the cosmic chi' blood are not in an active flow, it means there is a problem with life. What is in the greatest need in such a case is the sincere human mind that follows the principle of cosmic life. It is like the sincerity found in the pasonri singer, who mellows all the hardships and difficulties in the world out and create songs out of them like the shadow. It is the pansori singer's shadow that changes the universe. It is only when the extreme force of human mind communicates with that of the universe that the cosmic life or cosmic life culture can be created. In that sense, it is urgent to create life out of the universe inside me and create a universe out of all life in and outside me. It is such a grave plan in human history in that it involves finding the "Sanal" which is the core of life living hidden inside the body whose life force gradually goes away or inside the universe, and creating the culture of Bokseung in which it bursts out. The most important thing in life is the flow, and the mankind is currently standing in the life flow of the massive universe's chaosmos. The greatest task the mankind is currently faced with is to think over how to deal with the period of Big Chaos in the massive universe's chaosmos reversely and establish the cosmic life culture anew.

Effect of feeding mixed microbial culture fortified with trace minerals on ruminal fermentation, nutrient digestibility, nitrogen and trace mineral balance in Sheep

  • Kwak, W.S.;Kim, Y.I.;Choi, D.Y.;Lee, Y.H.
    • Journal of Animal Science and Technology
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    • 제58권5호
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    • pp.21.1-21.8
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    • 2016
  • Background: The aim of the present study was to determine the effects of feeding trace mineralsfortified mixed microbial culture (TMC) on ruminal fermentation, nutrient digestibility, blood electrolyte status, nitrogen balance, and trace mineral balance in sheep. Methods: Mixed microbes [0.6 % (v/w) of Enterobacter sp., Bacillus sp., Lactobacillus sp., and Saccharomyces sp.] were cultured with 99 % feedstuffs and 0.4 % trace minerals including zinc and copper for ensiling. Six sheep (a mean body weight of $46.5{\pm}1.2kg$) were fed two diets: a control diet (concentrate mix and rye straw) and an experimental diet (a control diet + 3.1 % TMC). Results: TMC feeding did not induce negative effects on ruminal fermentation, nutrient digestibility, blood electrolytes, and nitrogen balance in sheep. Feeding with TMC increased the intake of trace minerals (p < 0.05) and did not affect absorption of trace minerals in the whole digestive tract. Feeding with TMC increased fecal excretion and absorbable intake, and retention of zinc and copper (p < 0.05) by 71 % and 77 %, respectively. Conclusion: Feeding with TMC resulted in higher zinc and copper bioavailability and retention without any adverse effects on sheep performance.

전혈, 림프구와 백혈구 연층 각각의 방사성 동위원소 체외 조사 후 배양을 이용한 생물학적 선량측정 (Biological Dosimetry of In Vitro Irradiation with Radionuclides : Comparison of Whole Blood, Lymphocyte and Buffy Coat Culture)

  • 김종호;이동수;최창운;정준기;이명철;고창순;김종순;김희근;강덕원;송명재
    • 대한핵의학회지
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    • 제29권1호
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    • pp.125-132
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    • 1995
  • The purpose of this study was to establish mononuclear cell cultures such as lymphocytes or buffy coat for the biological dosimetry of in vitro Irradiation of the radionuclide Tc-99m in order to exclude the effect of residual doses seen in the cultures of whole blood. Biological do simetry of Tc-99m on cultured mononuclear cells at doses ranging from 0.05 to 6.00 Gy, by scoring unstable chromosomal aberrations(Ydr) observed in cultured lymphocytes, were performed using peripheral venous blood of healthy normal person. The results showed that; (1) In vitro irradiation of radioisotope in separated lymphocyte or buffy coat showed trace amount of residual doses of isotope after washing. Residual doses of isotopes are increased in proportion to exposed time and irradiated dose without difference between I-131 and Tc-99m. (2) We obtained these linear-quadratic dose response equations in lymphocyte and buffy coat culture after in vitro irradiation of Tc-99m, respectively (Ydr = 0.001949 $D^2$ +0.006279D + 0.000185; Ydr= 0.002531 $D^2$-0.003274 D+0.003488). In conclusion, the linear quadratic dose-response equation from in vitro irradiation of Tc-99m with lymphocyte and buffy coat culture was thought to be useful for assessing Tc-99m induced biological effects. And mono-nuclear cell cultures seem to be the most appropriate experimental model for the assessment of biological dosimetry of internal irradiation of radionuclides.

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Selection of Potential Virulence Factors Contributing to Streptococcus suis Serotype 2 Penetration into the Blood-Brain Barrier in an In Vitro Co-Culture Model

  • Liu, Hongtao;Zhu, Seng;Sun, Yingying;Li, Na;Gu, Jingmin;Sun, Changjiang;Feng, Xin;Han, Wenyu;Jiang, Jianxia;Lei, Liancheng
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.161-170
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    • 2017
  • Meningitis caused by Streptococcus suis serotype 2 (S. suis 2) is a great threat to the pig industry and human health. Virulence factors associated with the pathogenesis of meningitis have yet to be clearly defined, even though many potential S. suis 2 virulence factors have been identified. This greatly hinders the progress of S. suis 2 meningitis pathogenesis research. In this study, a co-culture blood-brain barrier (BBB) model was established using primary porcine brain microvascular endothelial cells and astrocytes, and the whole genome library of S. suis 2 was constructed using phage display technology. Finally, a total of 14 potential virulence factors contributing to S. suis 2 adherence to and invasion of the BBB were selected by analyzing the interactions between the phage library and the co-culture model. Twelve of these factors have not been previously reported in meningitis-related research. The data provide valuable insight into the pathogenesis of S. suis 2 meningitis and potential targets for the development of drug therapies.