• Title/Summary/Keyword: Active Cell

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EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Lee, Sung-Gyoon;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.511-519
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    • 2006
  • Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN DENTAL PAPILLA-DERIVED CELLS (배양된 치유두 유래세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Choi, Mun-Jeoung;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.4
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    • pp.279-288
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    • 2007
  • In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.

Active Polysaccharide and Immune Enhancement of Ganoderma lucidum Mycelium Cultured in Mushroom Complete Medium Supplemented with Ginseng Extract (수삼추출물 첨가 mushroom complete medium에서 배양된 영지버섯 균사체의 면역증진 효과 및 활성다당류)

  • Kim, Hoon;Jeong, Jae-Hyun;Jeong, Heon-Sang;Hwang, Jong-Hyun;Yu, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.633-640
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    • 2011
  • After Ganoderma lucidum was cultured in mushroom complete medium (MCM) supplemented with ginseng extract (GE), crude polysaccharide (GL-GE-CP) was fractionated from mycelium. Among GL-GE-CP from mycelium in MCM supplemented with 5, 10, and 15% GE (v/v ratio of MCM to GE), GL-GE-15-CP (15% GE) most significantly enhanced macrophage stimulation and intestinal immune system modulating activity compared with GL-CP in MCM without GE. When GL-GE-15-CP was further fractionated on DEAE-Sepharose CL-6B, GL-GE-15-CP-II displayed more potent activity than subfractions from GL-CP on macrophage stimulation, interleukin-12 production, and intestinal immune system modulation (1.75-, 5.68-, and 1.76-fold, respectively). Anti-metastasis effect against colon 26-M3.1 carcinoma cells was also enhanced by GL-GE-15-CP-II (72.8% inhibition). In addition, GL-GE-15-CP-II contained neutral sugar (83.00%) and uronic acid (9.11%), and consisted of Ara, Man, Gal and Glc (molar ratio of 0.39:0.50:0.75:1.00). Furthermore, GE supplementation helped to enhance the immunomodulation in G. lucidum, and it is assumed that neutral polysaccharides play an important role.

Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

A Study on the Stability and Moisturizing Effect for the Cream Containing Castanea crenata Leaf Extract (밤나무 잎 추출물 함유 크림의 안정성 및 피부 보습효과에 관한 연구 (2))

  • Kim, Jin-Young;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.4
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    • pp.301-307
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    • 2009
  • In the previous study, we evaluated and reported about the anti-oxidative activities of extract/fraction of Castanea crenata leaf. Extract/fraction of Castanea crenata leaf showed excellent free radical scavenging activity, cell protective activity and inhibitory activity on tyrosinase and elastase. In this study, in order to investigate the stability of cream containing 0.2 % Castanea crenata ethyl acetate fraction. pH, viscosity, and absorbance were measured under 4 different temperature ($4^{\circ}C$, $20^{\circ}C$, $37^{\circ}C$, $45^{\circ}C)$ and under the sun light at 2 weeks intervals for the 8 weeks. The variations on pH and viscosity of all experimental creams were similar to control cream. The absorbance variation of extract from experimental cream at 353 nm was in the order: under the sun > $45^{\circ}C$ > $37^{\circ}C$ > $20^{\circ}C$ > $4^{\circ}C$. It shows that ethyl acetate fraction in the cream can be oxidized under the sun. The bad smell and discoloration were not shown. Also, physical changes as creaming and cohesion were not shown. Also, transepidermal water loss (TEWL) and water contents in skin were measured. The cream containing Castanea cranata leaf extract was applied to the right lower arm. After 120 min, TEWL of parts was decreased as 29.7 % (experimental cream) and 5.4 % (control cream) respectively. And the water contents in skin were increased 22.6 % (experimental cream) and 24.7 % (control cream) respectively. It was confirmed that a cream containing ethyl acetate fraction of Castanea crenata leaf shows the superior moisturizing effect. The results showed that Castanea crenata leaf extract could be used as a new active ingredient for anti-aging cosmeceuticals.

o-Dihydroxyisoflavone Derivatives from Highly Aged Korean Fermented Soybean Paste by Jang Yang Process and Its Biological Activity (장양(藏釀) 기술이 사용된 전통 고숙성 된장에서 유래한 o-Dihydroxyisoflavone 유도체의 생리활성)

  • Kim, Dong-Hyun;Park, Jun-Seong;Park, Nok-Hyun;Moon, Eun-Jeong;Yu, Sun-Hye;Kim, Duck-Hee;Kim, Han-Kon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.3
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    • pp.203-208
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    • 2009
  • Doenjang (Korean fermented soybean paste) is a unique fermented food in Korea. It has been traditionally manufactured from soybeans, by Jang Yang process. We focused on the newly formed compound in highly aged Doenjang and its biological activity. One new o-dihydroxyisoflavone, 7,3',4'-trihydroxyisoflavone and two known o-dihydroxyisoflavone derivatives were isolated from 5-year-old Doenjang and evaluated as potent antioxidant and whitening effect by comparing with other known isoflavone. 7,8,4'-Trihydroxyisoflavone (compound 1), 7,3',4'-trihydroxyisoflavone (compound 2) and 6,7,4'-trihydroxyisoflavone (compound 3) inhibited DPPH (diphenyl-1-picrylhydrazyl) formation by 50 % at a concentration of $21.5{\pm}0.2$, $28.7{\pm}0.4$ and $32.6{\pm}0.6$ ($IC_{50}$) respectively, whereas daidzein showed weak DPPH radical scavenging activity. In superoxide scavenging effect were measured in one assay. Compound 1 ($IC_{50}=18.10{\pm}0.2{\mu}M$) and 2 ($IC_{50}=10.54{\pm}0.4{\mu}M$) show significant inhibitory activity and greater effect than L-ascorbic acid. But compound 3 and daidzein showed lower inhibition activity. Also, o-dihydroxyisoflavone derivatives evaluated as potent inhibitors on tyrosinase activity and melanin formation in melan-a cells. Compound 1 ($IC_{50}=11.21{\pm}0.2{\mu}M$), compound 2 ($IC_{50}=5.23{\pm}0.6{\mu}M$) exhibited significant inhibitory effect on tyrosinase activity. Furthermore, those compounds are significantly suppressed the cellular melanin formation by 50 % at a concentration of $12.23{\pm}0.7{\mu}M$ (1) and $7.83{\pm}0.7{\mu}M$ (2). This result suggests that 7,3',4'-trihydroxyisoflavone from highly aged Doenjang could be used as an active ingredient for cosmetics.

The Effect of Sodium Chloride on the Quality of Cheese and Upcoming Technologies for Manufacturing Reduced-Sodium Cheeses: A Review (Sodium Chloride가 치즈의 품질에 미치는 영향과 저염치즈 개발 기술: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Dong-Hyeon;Kim, Hong-Seok;Song, Kwang-Young;Jeong, Dong-Gwan;Kim, Soo-Ki;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.1
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    • pp.47-57
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    • 2015
  • Sodium is an essential nutrient with very important functions, including regulation of the extracellular fluid volume and active transport of molecules across the cell membranes. Since high levels of dietary sodium are associated with a high prevalence of hypertension, prehypertension, and other adverse effects on health, many national and international health organizations (WHO, FAO, etc.) recommend that sodium intake should be significantly decreased. In developed nations, cheese products, from among many processed foods, can cause high salt intake. Hence, there is an urgent need to reduce the content of salt in cheese processing, using various substitutes of sodium chloride (NaCl). In general, salt (NaCl) has been used as a food preservative to limit and (or) kill the growth of foodborne pathogens and spoilage organisms by decreasing the water activity, and to improve texture and flavor. To maintain public health, the salt content in cheese should be decreased without influencing the physicochemical properties of cheese. Therefore, the objective of this review is to outline the upcoming technologies used to reduce the salt content in different types of cheese using various substitutes.

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Staging of Esophageal Cancer Using Positron Emission Tomography : Comparing to Computed Tomography (양전자방출단층촬영술(PET)을 이용한 식도암 환자의 병기 결정 -전산화단층촬영술(CT)과의 비교-)

  • 심영목;박승준;김병태;김성철
    • Journal of Chest Surgery
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    • v.32 no.4
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    • pp.388-393
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    • 1999
  • Background: Correct preoperative staging of esophageal cancer is a prerequisite for adequate treatment. We prospectively compared the accuracy of positron emission tomography (PET) with [fluorine-18]FDG in the staging of esophageal cancer to that of computed tomography (CT). Material and Method: The findings of FDG PET and of chest CT including lower neck and the upper abdomen of 20 biopsy-proven squamous cell carcinoma patients (male, 19; female, 1; mean age, 61) were compared with the pathologic findings obtained from a curative esophagectomy with lymph node dissection. Result: The sensitivities of FDG PET and CT for diagnosis of primary tumor were the same, 90.0% (18/20). Both FDG PET and CT failed to show the primary tumor in 2 of 20 patients; one had a 1cm sized carcinoma in situ and the other had T1 stage cancer. By using the results of the pathologic examinations of 193 removed lymph node groups, we calculated the diagnostic sensitivities, specificities and accuracies of PET and CT (*$\chi$2 p < 0.005). Sensitivity** Specificity Accuracy* PET 55.6%(30/54) 97.1%(135/139) 85.5%(165/193) CT 13.0%(7/54) 98.6%(137/139) 74.6%(144/193) One of four patients with a false-positive for PEThad had active pulmonary tuberculosis. Among the 24 tumor involved lymph node groups, PET failed to show tumor metastasis in 5 lymph node groups abutting the tumor and in 14 lymph node groups located where the decay correction was not performed. Conclusion: Based on the above findings, it is suggested that [F-18]FDG-PET is superior to CT in the detection of nodal metastases and in the staging of patients with esophageal cancer.

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Study on the Antioxidant Effects of Nano-Selenium Microcapsule (Nano-Selenium Microcapsule의 항산화에 관한 연구)

  • Jeong, Hun;Yoo, Il-Su;Kim, Kyung-Sun;Lee, Soon-Young;Mun, Yeun-Ja;Jeon, Byoung-Kook;Ryu, Moon-Hee;Choi, Kyung-Soon
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.564-569
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    • 2012
  • Selenium was initially considered toxic to humans, but it was then discovered that selenium is essential for normal life processes. Selenium plays important roles in antioxidants. It is expected that chitosan microcapsules containing nano-selenium will be able to be used as a key material in bio-medical and cosmetic applications. The high concentration of chitosan derivatives guarantees increased antioxidative activity. Both inorganic and organic forms of selenium can be nutritional sources. The antioxidant properties of selenoproteins help prevent cellular damage from free radicals. The objective of this experiment was to study the antioxidative activity of chitosan nano-selenium. Our experiments were divided into five groups, in the presence of various concentrations(0.1%, 0.3%, 0.5%, 0.7%, and 0.9%) of chitosan. We performed an assessment of the antioxidant properties and cytotoxicity of respective concentrations of chitosan nano-selenium. The antioxidant activity was examined by the free radical scavenging activity on 1,1-diphenyl-2-picrylhydrazyl(DPPH) assay. The cytotoxicity effect was measured by means of 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. As a result, the electron donating abilities of 0.1%, 0.3%, 0.5%, 0.7%, and 0.9% of chitosan nano-selenium exhibited effective andioxidant scavenging activity at 12.5 ${\mu}g/m{\ell}$ against DPPH radicals. 0.3% chitosan nano-selenium did not show cytotoxicity on human keratinocytes. In general, the cytotoxicity of 0.1% and 0.9% chitosan nano-selenium showed the lowest effects. Though low cytotoxicity of 0.5% and 0.7% chitosan nano-selenium exhibited 29.67% and 38.4% against human keratinocytes on adding 100 ${\mu}g/m{\ell}$ and 50 ${\mu}g/m{\ell}$, respectively, cell vitality was recovered with 200 ${\mu}g/m{\ell}$. These findings support the notion that chitosan nano-selenium may be useful as a new active ingredient source for bioactive compounds.

Superoxide Generation by Blood Monocyte and Pulmonary Alveolar Macrophage in Patients with Pulmonary Tuberculosis (폐결핵환자의 폐포대식세포 및 말초혈액내 단구세포에서 분비하는 과산화음이온의 비교 관찰)

  • Song, Jeong-Sup;Lee, Suk-Young;Jang, Jie-Jung;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.1
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    • pp.11-19
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    • 1994
  • Background: Mycobacterium tuberculosis is a facultative intracellular pathogen which persists and multiplies within macrophage. Competent cell mediated immunity by cooperation of both T lymphocyte and macrophage of the host is required to kill the Mycobacterium tuberculosis. But a precise understanding of the pathogenesis of tuberculosis infection in pulmonary alveolar macrophage has not been achived. Research on the macrophage's basic microbicidal mechanism has elucidated the importance of oxygen-dependent or oxygen-independent components. Oxygen dependent processing begins with the reduction of oxygen by NADPH oxidase and generation of superoxide. In this study, the oxidative metabolic status of blood monocyte and pulmonary alveolar macrophage in patients with active pulmonary tuberculosis was accessed and compared with that of healthy control subjects to know whether there was a basic difference in superoxide generation by mononuclear cells between two groups. Methods: Pulmonary alveolar macrophage was purified after performing BAL(bronchoalveolar lavage) through the bronchi of infected lesion by plastic adhesion method. Blood monocyte was purified by Ficoll-Hypaque method. Superoxide generation by blood monocyte and pulmonary alveolar macrophage was measured by ferricytochrome-C reduction method after either stimulated with PMA(phorbol myristate acerate) or non-stimulated states. We also measured the effect of pulmonary tuberculosis patient's serum on superoxide generation by monocyte. Results: 1) Generation of superoxide by alveolar macrophage obtained from patients with pulmonary tuberculosis was little higher than those of controls, and PMA enhanced the generation of 2) Generation of superoxide by blood monocyte obtained from patients with pulmonary tuberculosis was little higher than those of control(p>0.05), and PMA more enhanced the generation of superoxide in patientswith pulmonary tuberculosis than those in controls(p<0.02). 3) Patient's serum enhanced the generation of superoxide by blood monocyte obtained from patients with pulmonary tuberculosis and controls, but not in the case of PMA stimulated blood monocyte. Conclusion: The present study suggest that the phenomenon of M.tuberculosis escape the microbicidal action of macrophage was not result of suppressed superoxide generation by blood monocyte and pulmonary alveolar macrophage, rather there might be a factor to stimulate the generation of superoxide by blood monocyte in pulmonary tuberculosis patient serum, but the comparision with effect of control's serum on superoxide generation needs further elucidation.

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