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An International Collaborative Program To Discover New Drugs from Tropical Biodiversity of Vietnam and Laos

  • Soejarto, Djaja D.;Pezzuto, John M.;Fong, Harry H.S.;Tan, Ghee Teng;Zhang, Hong Jie;Tamez, Pamela;Aydogmus, Zeynep;Chien, Nguyen Quyet;Franzblau, Scott G.;Gyllenhaal, Charlotte;Regalado, Jacinto C.;Hung, Nguyen Van;Hoang, Vu Dinh;Hiep, Nguyen Tien;Xuan, Le Thi;Hai, Nong Van;Cuong, Nguyen Manh;Bich, Truong Quang;Loc, Phan Ke;Vu, Bui Minh;Southavong, Boun Hoong
    • Natural Product Sciences
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    • v.8 no.1
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    • pp.1-15
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    • 2002
  • An International Cooperative Biodiversity Group (ICBG) program based at the University of Illinois at Chicago initiated its activities in 1998, with the following specific objectives: (a) inventory and conservation of of plants of Cuc Phuong National Park in Vietnam and of medicinal plants of Laos; (b) drug discovery (and development) based on plants of Vietnam and Laos; and (c) economic development of communities participating in the ICBG project both in Vietnam and Laos. Member-institutions and an industrial partner of this ICBG are bound by a Memorandum of Agreement that recognizes property and intellectual property rights, prior informed consent for access to genetic resources and to indigenous knowledge, the sharing of benefits that may arise from the drug discovery effort, and the provision of short-term and long-term benefits to host country institutions and communities. The drug discovery effort is targeted to the search for agents for therapies against malaria (antimalarial assay of plant extracts, using Plasmodium falciparum clones), AIDS (anti-HIV-l activity using HOG.R5 reporter cell line (through transactivation of the green fluorescent protein/GFP gene), cancer (screening of plant extracts in 6 human tumor cell lines - KB, Col-2, LU-l, LNCaP, HUVEC, hTert-RPEl), tuberculosis (screening of extracts in the microplate Alamar Blue assay against Mycobacterium tuberculosis $H_{37}Ra\;and\;H_{37}Rv),$ all performed at UIC, and CNS-related diseases (with special focus on Alzheimer's disease, pain and rheumatoid arthritis, and asthma), peformed at Glaxo Smith Kline (UK). Source plants were selected based on two approaches: biodiversity-based (plants of Cuc Phuong National Park) and ethnobotany-based (medicinal plants of Cuc Phuong National Park in Vietnam and medicinal plants of Laos). At mc, as of July, 2001, active leads had been identified in the anti-HIV, anticancer, antimalarial, and anti- TB assay, after the screening of more than 800 extracts. At least 25 biologically active compounds have been isolated, 13 of which are new with anti-HIV activity, and 3 also new with antimalarial activity. At GSK of 21 plant samples with a history of use to treat CNS-related diseases tested to date, a number showed activity against one or more of the CNS assay targets used, but no new compounds have been isolated. The results of the drug discovery effort to date indicate that tropical plant diversity of Vietnam and Laos unquestionably harbors biologically active chemical entities, which, through further research, may eventually yield candidates for drug development. Although the substantial monetary benefit of the drug discovery process (royalties) is a long way off, the UIC ICBG program provides direct and real-term benefits to host country institutions and communities.

Ultrafiltration for Quality Improvement of Apple Wine (한외여과공정을 이용한 사과주의 품질개선)

  • Chung, Jae-Ho;Mok, Chul-Kyoon;Lim, Sang-Bin;Park, Young-Seo
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.201-206
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    • 2003
  • An apple wine was prepared by fermentation at $25^{\circ}C$ for 2 weeks using Saccha개myces cerevisiae KCCM 12224, followed by aging at $15^{\circ}C$ for 14 weeks, and its physicochemical and microbiological changes were investigated. The viable bacterial cell numbers, increased from $1.4{\times}10^3\;CFU/ml$ at the beginning of fermentation, to $2.8{\times}10^6\;CFU/ml$ after 2 weeks, but decreased to $1.0{\times}10^5\;CFU/ml$ after aging. The viable yeast cell numbers changed from $4.3{\times}10^4\;CFU/ml$ to $1.2{\times}10^7\;CFU/ml$ during the fermentation, and decreased to $1.2{\times}10^4\;CFU/ml$ after aging. Sugar content changed from $20.0^{\circ}Brix$ to $8.5{\circ}Brix$, and reducing sugar content was changed from 9.66% to 6.44%. Alcohol content and acidity increased to 7.0% and from 0.19% to 0.24%, respectively. No changes in acidity, pH, and sugar content were observed during the aging, but reducing sugar and solid contents decreased. When apple wine was fultered through $0.45\;{\mu}m$ nitrocellulose membrane followed by various ultrafiltration membranes with different molecular weight cut-off values, the initial flux $(121.2\;liter/m^2/h)$ and the average flux of Biomax 100k membrane were the highest among the membranes used. These membrane filtration treatments resulted in complete removal of microorganisms as well as decrease in turbidity and solid content without changes in other chemical properties. No changes in the physicochemical properties of the apple wine and no microorganisms were detected during the storage at $156{\circ}C$ for 6 weeks.

Phytoplankton and Bacterioplankton in the Intertidal and Subtidal Waters in the Vicinity of Kunsan (군산부근 조간대 및 조하대역에서의 식물플랑크톤과 Bacterioplankton)

  • Lee, Won Ho;Lee, Gean Hyoung;Choi, Moon Sul;Lee, Da Mi
    • 한국해양학회지
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    • v.24 no.3
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    • pp.157-164
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    • 1989
  • Quantitative species distribution and primary productivity of phytoplankton were studied monthly from August, 1987 to July, 1988 along with the quantitative distribution of total heterotrophic bacterioplankton and three groups of physiologically chracteristic bacterioplankton in the intertidal and subtidal waters off Kum River Estuary, Yellow Sea. A total of 121 phytoplankton taxa including 102 diatoms occurred, and cell concentration ranged from 15 to 5451 (cells/ml). The great spatio-temporal variations of the number of phytoplankton species and cell concentration well reflected the environmental differences between the intertidal and subtidal waters. Primary productivity (in Piopt, mgC/$m^3$/hr) ranged from 0.6 to 27.3. Just after the phytoplankton bloom (March) Piopt was very low in April at station 1, where amylolytic bacterioplankton also showed quite low population density. The peaks of primary productivity were not always coincided with those of phytoplankton standing crop. The ratio of Piopt's between samples well indicated the environmental differences between the intertidal and subtidal waters. Little characteristic trend was found in the scatter diagrams of phytoplankton standing crop along the population densities of total heterotrophic bacterioplankton and the three groups of physiologically characteristic bacterioplankton. In summer the phytoplankton standing crop was minimum in contrast with the high population density of bacterioplankton, which implies the influx of much allochthonous orgainc matter from Kum River. The scatter diagrams of Piopt along bacterioplankton population density revealed some phenomena there. Piopt had highly positive correlation with the population density of amylolytie bacterioplankton($R^2$=0.84) and that of lipolytic bacterioplankton($R^2$=0.70) while total heterotrophic bacterioplankton and proteolytic bacterioplankton had lesser correlations with Piopt. From the regression lines the increase of unit Piopt (mgC/$m^3$/hr) in the study area was calculated to mean the increase of $9.0{\times}10$ cells/ml and $8.0{\times}10$ cells/ml of amylolytic bacterioplankton and lipolytic bacterioplankton, respectively.

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Utility of FDG-PET in Solitary Pulmonary Nodules and the Relationship Between Standardized Uptake Values of PET and Serum Glucose (폐 결절에서 FDG-PET의 유용성과 표준섭취계수와 혈당농도의 상관관계)

  • Kim, Kyu Sik;Lim, Sung Chul;Ko, Young Chun;Park, Kyung Ha;Ju, Jin Young;Jo, Kae Jung;Kim, Soo Ok;Oh, In Jae;Kim, Yu Il;Kim, Young Chul;Kim, Sung Min;Song, Ho Chun;Bom, Hee Seung;Park, Kyung Ok
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.6
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    • pp.589-596
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    • 2003
  • Background : The solitary pulmonary nodule(SPN) presents a diagnostic dilemma to the physician and the patients in the our nation with high incidence of tuberculoma. We could not exclude whether the SPN was benign or malignant by the change of the size at chest radiograph and findings of chest CT. Recently, positron emission tomography(PET) have been tried as the differential diagnostic method of SPN. We evaluated the efficacy of PET for differentiating malignant from benign SPN and the relationship between standardized uptake values(SUV) of PET and serum glucose. Method : Between January 2001 and July 2002, sixty-one patients with pulmonary nodule were examined by the chest CT and PET. The SPN has been finally diagnosed by the transthorasic needle aspiration and biopsy, bronchoscopic biopsy, and open lung biopsy. Results : Forty eight patients had a malignant nodule(23 squamous cell lung carcinoma, 16 adenocarcinoma, 9 small cell lung cancer) and thirteen patients had a benign nodule(3 tuberculoma, 9 inflammatory granuloma, 1 cryptococcosis). The mean size of malignant and benign nodule was 40.6 mm and 20.0 mm, respectively. All malignant nodules showed a marked increase in 18 fluorodeoxyglucose (FDG) uptake. Mean SUV of malignant was $9.52{\pm}5.20$ and benign nodule was $1.61{\pm}3.60$. There were false positive cases with an increase in 18-FDG uptake (2 tuberculoma, 1 inflammatory granuloma). The SUV of malignant nodule in diabetes patients has no difference in non diabetes patients($9.10{\pm}4.51$ vs $9.65{\pm}5.46$). The sensitivity and specificity of the PET scan for SPN were 100%, 77%, respectively. The positive and negative predictive values were 94% and 100%. Conclusion : PET scanning showed highly accurate result in differentiating the malignant and benign SPN. There were no significant differences between the SUV and serum glucose in the patients with lung cancer.

Rice Fermentation by Korean Amylolytic Bifidobacterium spp. (한국인유래의 Amylolytic Bifidobacterium 에 의한 쌀발효)

  • Park, Jong-Hyun;Song, Hey-Kyung;Ahn, Jun-Bae;Ji, Geun-Eok;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.581-587
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    • 1997
  • For bifidus fermentation food, gelatinized rice solution was fermented without liquefaction/saccharification by amylolytic Bifidobacterium spp. isolated from Korean. Eighteen amylolytic Bifidobcterium on the starch agar were isolated from 38 Korean and four strains were finally selected as good amylase producers. The most enzyme-producing strain of Bif. sp. FBD-12 secreted extracellular amylase of 0.17 U/mg and intracelluar amylase of 1.8 U/mg. Three strains of Bif. sp. FBD-12, Bif. sp. FBD-16 and Bif. sp. FBD-17 also showed good growth on pH controlled media by HCI/acetic acid to pH 5.0 while Bif. sp. FBD-6 was not so tolerant that viable cell counts reduced to $10^2\;CFU/mL$ times on the media. Initial cell number of $10^6\;CFU/mL$ for those strains reached to $10^9\;CFU/mL$ on the rice medium supplemented with yeast extract (0.2%) and cysteine (0.05%). Ascorbic acid instead of cysteine was added to the medium for improving off-flavour and the best growth was shown at 0.1% addition. Isolated soybean proteins (ISP) of 3% accelerated the growth of the strains. Maximum count of $10^9\;CFU/mL$ reached within 12 hour fermentation on the rice medium with ascorbic acid and isolated soybean protein instead of 32 hours on the cysteine medium, and total acidity increased from 0.5% to 1% on each media. Reducing sugar in the ascorbic acid/ISP cultures generally increased especially 2 mg/mL to 15.5 mg/mL for Bif. sp. FBD-6. From sensory evaluation, the products showed good acceptability so that it suggested possibility of development of bifidus-fermented rice food.

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Diagnostic Value of Serum Cytokeratin 8, 18 and 19 in Lung Cancer (폐암에서 혈중 Cytokeratin 8, 18, 19의 진단적 가치)

  • Choi, Chang Min;Kim, Woo Jin;Oh, Jin Young;Kang, Young Ae;Yoo, Chul Gyu;Lee, Choon Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.4
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    • pp.388-394
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    • 2003
  • Background : Monoclonal antibodies directed against well-known epitopes on cytokeratin (CK) 8, 18 and 19 (Monototal) have been used in the development of a new diagnostic tool for lung cancer. In the mid-1990s, CK 19 fragments (Cyfra 21-1) became popular and widely used for such diagnosis. This is the first study specifically designed to compare these two markers. Method : The serum levels of CK 8, 18 and 19 were measured using two-site monoclonal/polyclonal immunoradiometric assay kit in 57 healthy adults and 289 patients who were admitted to Seoul National University Hospital from May to September, 2002. The lung cancer group comprised 129 primary lung cancer patients; 116 with non-small cell lung cancer(NSCLC) and 13 with small cell lung cancer (SCLC). The control group comprised 160 non-malignant pulmonary lung disease patients and 57 healthy adults. A total of 166 twin Monototal and Cyfra 21-1 serum assays were obtained; 76 with lung cancer, 70 with non-malignant pulmonary lung disease and 20 healthy adults. Results : The mean serum value of Monototal was $412.47{\pm}455.45U/L$ in NSCLC, $237.08{\pm}145.15U/L$ in SCLC, $126.54{\pm}95.72U/L$ in non-malignant pulmonary lung disease, and $63.68{\pm}31.66U/L$ in healthy adults. The serum values of the lung cancer groups were significantly higher than those of the control group (p<0.01). Using a cut off value of 188U/L, sensitivity and specificity was 66.4% and 81.9% in NSCLC, and 43.8% and 81.9% in SCLC, respectively. The serum levels of CK 8, 18 and 19 were higher in advanced NSCLC than in early stage disease. Conclusion : The serum levels of CK 8, 18 and 19 may be useful in the diagnosis of NSCLC.

Two Crystal Structures of the Vacuum-Dehydrated Fully $Ag^+$-Exchanged Zeolite X ($Ag^+$ 이온으로 완전히 치환되고 탈수된 두개의 제올라이트 X의 결정구조)

  • Jang, Se Bok;Park, Sang Yun;Song, Seong Hwan;Jeong, Mi Suk;Kim, Yang
    • Journal of the Korean Chemical Society
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    • v.40 no.7
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    • pp.474-482
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    • 1996
  • Two crystal structures of the vacuum dehydrated $Ag^+$-exchanged zeolite X have been determined by single-crystal X-ray diffraction techniques in the cubic space group Fd3 at 21(1)$^{\circ}C$ (a=24.922(1)${\AA}$ and a=24.901(1)${\AA}$, respectively). Each crystal was ion exchanged in flowing streams of aqueous $AgNO_3$ for three days. The first crystal was dehydrated at 300$^{\circ}C$ and $2{\times}10^{-6$torr for two days. The second crystal was similarly dehydrated at 350$^{\circ}C$. Their structures were refined to the final error indices, $R_1=0.095\;and\;R_2=0.092$ with 227 reflections, and $R_1=0.096\;and\;R_2=0.087$ with 334 reflections, respectively, for which I > 3${\sigma}$(I). In the first crystal, Ag species are found at five different crystallographic sites: sixteen $Ag^+$ ions fill the site I, the center of the double 6-ring, thirty-two Ag0 atoms fill the I' site in the sodalite cavities opposite double six-rings, seventeen $Ag^+$ ions lie at the 32-fold site II' inside the sodalite cavity at the single six-oxygen ring in the supercage, fifteen Ag+ ions lie at the 32-fold site II, in the supercage, and the remaining twelve $Ag^+$ ions lie at site III' in the supercage at a little off two-fold axes. In the second crystal, all Ag species are located similarly as crystal 1; 16 at site I, 28 at site I', 16 at site II, 16 at site II', 6 at site III and 6 at site III'. Total 88 silver species were found per unit cell. The remaining four Ag atoms were migrated out of the zeolite framework to form small silver crystallites on the surface of the zeolite single crystal. In the first structure, the numbers of Ag atoms per unit cell are approximately 32.0 and these may form tetrahedral $Ag_4$ clusters at the centers of the sodalite cavities. The probable four-atom cluster is stabilized by coordination to two $Ag^+$ ions. The Ag-Ag distance in the cluster, ca. 3.05 ${\AA}$, is a little longer than 2.89 ${\AA}$, Ag-Ag distance in silver metal. At least two six-ring $Ag^+$ ions on sodalite cavity (site II') must necessarily approach this cluster and this cluster may be viewed as a distorted octahedral silver cluster, (Ag6)2+.

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Diagnostic Significance of TNF-$\alpha$ in Tuberculous and Non-Tuberculous Pleural Effusion (결핵성 및 비결핵성 흉막삼출액에서 TNF-$\alpha$ 농도의 진단적 의의)

  • Na, Hyun-Joo;Park, Seog-Chea;Kang, Kwang-Won;Park, Hyeong-Kwan;Kim, Young-Chul;Choi, In-Seon;Park, Kyung-Ok
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.611-620
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    • 1997
  • Objectives : The differentiation of tuberculous effusion from the other causes of exudative pleural effusion remained difficult even with aids of biochemical analyses and pleural biopsy. As the pathophysiology of tuberculous pleural effusion is an enhanced cell mediated immunity, Adenosine deaminase(ADA) and various eytokines including Inteferon-$\gamma$, tumor necrosis factor alpha(TNF-$\alpha$) are considered as useful diagnostic tools in differentiating exudative pleural effusion. The author would like to demonstrate the diagnostic usefulness of TNF-$\alpha$ in the differentiation of exudative pleural effusion, and compared the discriminating ability of TNF-$\alpha$ with ADA. Methods : Pleural fluids obtained from 80 patients (tuberculous : 39, malignant : 31, parapneumonic : 10) with exudate pleural effusions were processed for cell counts and biochemical analysis including ADA and TNF-$\alpha$. Results : Tuberculous pleural fluid showed higher levels of ADA and TNF-$\alpha$, $48.7{\pm}32.7U/L$ and $184.1{\pm}214.2pg/mL$ than that of non-tuberculous effusion $26.0{\pm}41.3U/L$ and $44.1{\pm}114.2pg/mL$, respectively (ADA, TNF-$\alpha$, p < 0.05, p < 0.01). Receiver operating characteristics(ROC) curves were generated for ADA and TNF-$\alpha$ and the best cut-off value for adenosine deaminase and TNF-$\alpha$were considered as 30U/L and 15pg/ml, respectively. Comparing the area under the ROC curves, there was no significant difference between ADA and TNF-$\alpha$. Conclusion : For the differential diagnosis of tuberculous pleural effusion from the other causes of exudative pleural effusions, TNF-$\alpha$ as well as ADA was considered as useful diagnostic method. However adding TNF-$\alpha$ to ADA has no further diagnotic benefit than ADA alone.

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Molecular Miology of the Poliovirus (폴리오바이러스의 분자생물학)

  • 최원상
    • Journal of Life Science
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    • v.7 no.4
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    • pp.392-401
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    • 1997
  • The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

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The Significance of IL-10, IL-12, IFN-$\gamma$ and ADA in Tuberculous Pleural Fluid (결핵성 흉수에서 IL-10, IL-12, IFN-$\gamma$, ADA 측정의 의의)

  • Jeon, Doo-Soo;Yun, Sang-Myung;Park, Sam-Seok;Lee, Hyo-Jin;Kim, Yun-Seong;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.301-310
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    • 1998
  • Background: Cell mediated immune response mediated by interaction between CD4+ T lymphocytes and macrophagies is thought to play an important role in tuberculous pleurisy. This interaction is dependent on the interplay of various cytokines. The immunologic response of tuberculous pleurisy is thought to depend on the balance between helper T cell(Th1) cytokine Interleukin-12, Interferon gamma and Th2 cytokine IL-4, IL-10. To understand immunologic mechanism in tuberculous pleurisy and evaluate diagnostic value of these cytokines, the concentrations of Th1 cytokine IL-12, IFN -$\gamma$ and Th2 cytokine IL-10 were measured in tuberculous pleurisy and malignant pleural effusion group. Material and Methods: The concentrations of IL-10, IL-12 and IFN-$\gamma$ were measured by ELISA method in pleural fluids and serums of 20 patients with tuberculous pleurisy and 20 patients with malignant pleural effusion ADA activities were measured by spetrophotomery in pleural fluids of both groups. Results: In tuberculous pleurisy, the mean concentrations of IL-10, IL-12 and IFN-$\gamma$ of pleural fluids showed $121.3{\pm}83.7$ pg/mL, $571.4{\pm}472.7$ pg/mL and $420.4{\pm}285.9$ pg/mL. These were significantly higher than that of serum, $21.2{\pm}60.9$ pg/mL, 194.5 pg/mL, $30.1{\pm}18.3$ pg/mL respectively(p< 0.01). In malignant pleural effusion, the mean concentrations of IL-10, IL-12 and IFN-$\gamma$ of pleural fluids showed $88.4{\pm}40.4$ pg/mL, $306.5{\pm}271.1$ pg/mL and $30.5{\pm}54.8$ pg/mL respectively. Compared with that of serum ($43.4{\pm}67.2$ pg/mL, $206.8{\pm}160.6$ pg/mL, $14.6{\pm}3.3$ pg/mL), only IL-10 was significantly higher (p<0.001), but IL-12, IFN-$\gamma$ were not significant. In tuberculous pleural effusion compared with malignant pleural effusion, the concentration of IL-12, IFN-$\gamma$, ADA were significantly higher (p=value 0.046, <0.001, <0.001), but IL-10 was not significant. For differential diagnosis of tuberculous pleurisy from malignant pleural effusion, using cut-off value of IL-12, IFN-$\gamma$, ADA as 300 pg/mL. 100 pg/mL, 45 U/L, the sensitivity/specificity were 60%/70%, 90%/87.5%, 85%/90% respectively. Conclusion: In tuberculous pleurisy, IL-10, IL-12 and IFN-$\gamma$ were selectively concentrated highly in pleural space than serum. Compared with malignant pleural effusion, IL-12 and IFN-$\gamma$ were significantly higher, but IL-10 were not in tuberculous pleural effusion. The results suggest that Th1 pathway contributes to immune resistant mechanism in tuberculous pleurisy. IFN-$\gamma$ and ADA revealed useful methods of differential diagnosis in tuberculous pleurisy from malignant pleural effusion.

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