• Title/Summary/Keyword: Whole blood culture

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

  • Cheon, Seon-Hee;Song, Ho-Yeon;Lee, Eun-Hee;Oh, Hee-Jung;Kang, In-Sook;Cho, Ji-Yoon;Hong, Young-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.497-509
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    • 2002
  • Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.

Sister Chromatid Exchanges in Lymphocytes on Normal Human Blood Culture with Mercury chloride or Methylmercury Chloride (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))

  • Koh, Dai-Ha
    • Journal of Preventive Medicine and Public Health
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    • v.17 no.1
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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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Phytomitogen induced blast transformation of rabbit I. Effect of medium, phytomitogen and culture hours on the uptake of 3H-thymidine (Phytomitogen에 의한 토끼 임파구의 blast transformation I. 유사분열에 미치는 배지, 유사분열촉진물질 및 배양시간의 효과)

  • Kim, Jong-shu;Kim, Chung-hui
    • Korean Journal of Veterinary Research
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    • v.31 no.1
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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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Transnational Adoption and Beyond-Borders Identity: Jane Jeong Trenka's The Language of Blood (초국가적 입양과 탈경계적 정체성 -제인 정 트렌카의 『피의 언어』)

  • Kim, Hyunsook
    • Journal of English Language & Literature
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    • v.57 no.1
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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.

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

  • Kwon, Soon-Oh;Lam, Truong Quang;Her, Moon;Ahn, Dong-Chun;Park, Sang-Hee;Park, Mi-Yeoun;Lee, Young-Ju;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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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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    • v.15 no.5
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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.

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

  • Lee, Jae-bok
    • Cross-Cultural Studies
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    • v.21
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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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    • v.58 no.5
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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.

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

  • Kim, Jong-Ho;Lee, Dong-Soo;Choi, Chang-Woon;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon;Kim, Chong-Soon;Kim, Hee-Geun;Kang, Duck-Won;Song, Myung-Jae
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.1
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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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    • v.27 no.1
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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.