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Purification of Vibrio anguillarum Growth Inhibition Factor Produced by Bacillus amyloliquefaciens H41. (Bacillus amyloliquefaciens H41이 생산하는 Vibrio anguillarum 생육 저해인자의 정제)

  • Shin, Hyun-Chul;Chung, Kyung-Tae;Kim, Kwang-Hyun;Kim, Byung-Woo;Kwon, Hyun-Ju;Lee, Eun-Woo;Yum, Jong-Hwa;Rhu, Eun-Ju;Jeong, Yu-Jeong;Kim, Young-Hee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.789-795
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    • 2008
  • To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.

The Effects of Dietary Fermented Fruit Pomace and Angelica keiskei Koidz Pomace on Shelf Life, Cholesterol and Fatty Acid Composition in Broiler (발효 과일박 및 신선초박의 급여가 계육 내 지방산 조성, 콜레스테롤 및 저장 기간 중 지방산패도에 미치는 영향)

  • Kang, Hwan-Ku;Choi, Hee-Chul;Chae, Hyun-Suk;Na, Jae-Cheon;Bang, Han-Tae;Park, Sung-Bok;Kim, Min-Ji;Seo, Ok-Suk;Lee, Jee-Eun;Kim, Dong-Wook;Kim, Sang-Ho;Kang, Guen-Ho
    • Food Science of Animal Resources
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    • v.30 no.3
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    • pp.466-471
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    • 2010
  • This study investigated the effects of dietary supplementation of fermented apple pomace (FAP), fermented pear pomace (FPP), fermented orange pomace (FOP), and fermented Angelica keiskei Koidz pomace (FAKP) on performance, shelf life, fatty acid composition and cholesterol in broiler chickens. A total of 600, 1-day-old male broiler chicks (Cobb strain) were randomly divided into six groups with four replicates of 30 birds each. There were five treatment groups: control (C), FAP (1.0%, T1), FPP (1.0%, T2), FOP (1.0%, T3), and FAKP (1.0%, T4). The body weight of the broiler chickens fed FAP diet was higher (1,758 g) than the other treatments. There was no difference in the thiobarbituric acid reactive substances (TBARS) in chicken meat between all groups at days 1, 3, and 5 of storage, while the FAP-supplemented group displayed lower TBARS values at day 7. There was no significant difference in fatty acid composition between the groups but the cholesterol content of chicken meat was lower than the control groups. These results suggest the possibility that FAP could be used as a functional feed to improvement the quality performance of broiler chickens.

The Difference in Chemokine Expression in Airway Epithelial Cells According to the Virulence of Tubercle Bacilli (결핵균 독성 여부에 따른 기도 상피세포의 Chemokine 발현에 관한 연구)

  • Kwon, O-Jung;Kim, Ho-Joong;Kim, Jung-Hee;Kim, Ho-Cheol;Suh, Gee-Young;Park, Jeong-Woong;Park, Sang-Joon;Chung, Man-Pyo;Choi, Dong-Chull;Rhee, Chong-H.
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.729-741
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    • 1997
  • Background : We have recently reported that airway epithelial cells can produce RANTES and IL-8 in response to the stimulation of tubercle bacilli suggesting a certain role of airway epithelial cells in the pathogenesis of pulmonary tuberculosis. The pathogenesis of tuberculosis is determined by several factors including phagocytosis, immunological response of host, and virulence of tubercle bacilli. Interestingly, there have been reports suggesting that difference in immunological response of host according to the virulence of tubercle bacilli may be related with the pathogenesis of tuberculosis. We, therefore, studied the expressions and productions of RANTES and IL-8 in airway epithelial cells in response to tubercle bacilli(H37Rv, virulent strain and H37Ra, avirulent strain), in order to elucidate the possible pathophysiology of pulmonary tuberculosis. Methods : Peripheral blood monocytes were isolated from normal volunteers. Peripheral blood monocytes (PBM) were stimulated with LPS($10{\mu}g/ml$), H37Rv, or H37Ra($5{\times}10^5$ bacilli/well) along with normal control for 24 hours. A549 cells were stimulated with supernatants of cultured PBM for 24 hours. ELISA kit was used for the measurement of $TNF{\alpha}$ and IL-$1{\beta}$ production in supernatants of cultured PBM and for the measurement of RANTES and IL-8 in supernatants of cultured A549 cells. Northern blot analysis was used for the measurement of RANTES and IL-8 mRNA expression in cultured A549 cells. Results : $TNF{\alpha}$ and IL-$1{\beta}$ productions were increased in cultured PBM stimulated with LPS or tubercle bacilli(H37Rv or H37Ra) compared with the control. There was, however, no difference in $TNF{\alpha}$ and IL-$1{\beta}$ production between cultured PBM stimulated with H37Rv and H37Ra. RANTES and IL-8 expressions and productions were also increased in cultured A549 cells stimulated with LPS or tubercle bacilli compared with the control. RANTES and IL-8 mRNA expressions were significantly increased in cultured A549 cells stimulated with H37Ra-conditioned media(CM) compared with A549 cells stimulated with H37Rv-CM (p<0.05). However, there was no difference in RANTES and IL-8 productions between A549 cells stimulated with H37Rv-CM and H37Ra-CM. Conclusion : Airway epithelial cells can produce the potent chemokines such as RANTES and IL-8, in response to the stimulation of tubercle bacilli. These results suggest that airway epithelial cells may play a certain role in the pathogenesis of pulmonary tuberculosis. However, the role of airway epithelial cells in the pathogenesis of tuberculosis according to the virulence of tubercle bacilli was not clear in this study.

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Effect of 6-Hydroxydopamine (6-OHDA) on the Expression of Hypothalamus-Pituitary Axis Hormone Genes in Male Rats (수컷 흰쥐의 시상하부-뇌하수체 축 호르몬 유전자 발현에 미치는 6-Hydroxydopamine(6-OHDA)의 영향)

  • Heo, Hyun-Jin;Ahn, Ryun-Sup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.13 no.4
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    • pp.257-264
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    • 2009
  • A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD) due to its specific toxicity against dopaminergic (DA) neurons. Since DA signals modulate a broad spectrum of CNS physiology, one can expect profound alterations in neuroendocrine activities of both PD patients and 6-OHDA treated animals. Limited applications of 6-OHDA injection model, however, have been made on the studies of hypothalamuspituitary neuroendocrine circuits. The present study was performed to examine whether blockade of brain catecholamine (CA) biosynthesis with 6-OHDA can make any alteration in the transcriptional activities of hypothalamus-pituitary hormone genes in adult male rats. Three-month-old male rats (SD strain) were received 6-OHDA ($200{\mu}g$ in $10{\mu}\ell$ of saline/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. To determine the mRNA levels of hypothalamuspituitary hormone genes, total RNAs were extracted and applied to the semi-quantitative RT-PCRs. The mRNA levels of tyrosine hydroxylase (TH), the rate-limiting enzyme for the catecholamine biosynthesis, were significantly lower than those from the control group (control:6-OHDA=1:0.72${\pm}$0.02AU, p<0.001), confirming the efficacy of 6-OHDA injection. The mRNA levels of gonadotropin-releasing hormone (GnRH) and corticotropin releasing hormone (CRH) in the hypothalami from 6-OHDA group were significantly lower than those from the control group (GnRH, control:6-OHDA=1:0.39${\pm}$0.03AU, p<0.001; CRH, control:6-OHDA=1:0.76${\pm}$0.07AU, p<0.01). There were significant decreases in the mRNA levels of common alpha subunit of glycoprotein homones (Cg$\alpha$), LH beta subunit (LH-$\beta$), and FSH beta subunit (FSH-$\beta$) in pituitaries from 6-OHDA group compared to control values (Cg$\alpha$, control:6-OHDA=1:0.81${\pm}$0.02AU, p<0.001; LH-$\beta$, control:6-OHDA=1:0.68${\pm}$0.04AU, p<0.001; FSH-$\beta$, control:6-OHDA=1:0.84${\pm}$0.05AU, p<0.001). Similarly, the level of adrenocorticotrophic hormone (ACTH) transcripts from 6-OHDA group was significantly lower than that from the control group (control: 6-OHDA=1:0.86${\pm}$0.04AU, p<0.01). The present study demonstrated that centrally injected DA neurotoxin could downregulate the transcriptional activities of the two hypothalamus-pituitary neuroendocrine circuits, i.e., GnRH-gonadotropins and CRH-ACTH systems. These results suggested that hypothalamic CA input might affect on the activities of gonad and adrenal through modulation of hypothalamus-pituitary function, providing plausible explanation for frequent occurrence of sexual dysfunction and poor stress-response in PD patients.

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A Study on the Manufacturing of Soysauce by the Combined use of Aspergillus sojae and Bacillus subtilis (단백질분해세균(蛋白質分解細菌)을 병용(倂用)한 간장제조(製造)에 관(關)한 연구(硏究))

  • Kim, Ze-Uook;Cho, Sung-Hwan
    • Applied Biological Chemistry
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    • v.18 no.1
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    • pp.1-9
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    • 1975
  • In order to study on the preparation of improved soysauce, Koji(soybean and wheat mash fermented by the strain Aspergillus sojae), Koji-Natto(mixed with the various proportion of Koji to Natto; 8 : 2, 6 : 4, 4 : 6, 2 : 8) and Natto (prepared using the strain: Bacillus subtilis) were prepared as soysauce materials. Sensory evaluation for the soysauces made from above-mentioned soysauce materials and the contents of total nitrogen, amino nitrogen, ammonia nitrogen, reducing sugar, total acid and enzyme activity were determined. The results were summarized as follows: 1. Protease activity of Natto was over twice higher than that of Koji and amylase activity of Koji was three times higher than that of Natto. Koji-Natto showed successive increase of amylase activity and successive decrease of protease activity as Koji proportion was increased to Natto. 2. The contents of total nitrogen and amino nitrogen in all the soysauce mashes displayed linear increases or no changes of these contents as aging. The contents of total nitrogen and amino nitrogen in the mahes were higher as Natto proportion increase to Koji during the aging period. 3. In Natto soysauce mash and Koji-Natto soysauce mashes the contents of ammonia nitrogen were gradually increased until $1{\sim}2$ months and rapidly reduced after that period, while Koji soysauce mashes were continuatively increased. 4. In all the soysauce mash the reducing sugar increased preeminently until the two months and after then there was a rapid decrease as aging the contents of reducing sugar in the mash were higher as Natto proportion increased to Koji during aging period. 5. The total acid of Koji and Koji-Natto (8 : 2) soysauce mash showed gradual increase as aging, while contents of total acid in Natto and other Koji-Natto soysauce mashes increased preeminently until 50${\sim}$70 days aging, after then decreased. 6. The results of sensory evaluation for three-month-fermented soysauces showed the most excellent in Koji-Natto (6 : 4) and successive order in the soysauces consisting of Koji and Natto proportion: 8 : 2, 10 : 0, 4 : 6, 2 : 8, 0 : 10.

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Anti-Obesity Effects of Fermented Soybean Oils in 3T3-L1 Pre-Adipocytes and High Fat Diet-Fed C57BL/6J Mice (발효콩 유지의 3T3-L1 지방전구세포와 고지방식이를 급여한 C57BL/6J 생쥐에 대한 항비만 효과)

  • Kim, Seon-Woong;Kim, Nam-Seok;Oh, Mi-Jin;Kim, Ha-Rim;Kim, Min-Sun;Lee, Da-Young;Yoon, Suk-Hoo;Jung, Mun-Yhung;Kim, Hun-Jung;Lee, Chang-Hyun;Oh, Chan-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.3
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    • pp.279-288
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    • 2017
  • This study investigated the manufacturing of fermented soybean oil using a fermenting strain commonly processed for soybeans [Bacillus amyloliquefaciens (BA), Bacillus subtilis (BS), Lactobacillus acidophilus (LBA), and B. subtilis+L. acidophilus (BLO)] and evaluated its anti-obesity activities. Cytotoxicity of four kinds of fermented soybean oils was not observed in 3T3-L1 preadipocytes at 10 and $50{\mu}g/mL$. Triglyceride content was reduced by 20.6% in the BLO group at a treatment concentration of $50{\mu}g/mL$. The simultaneous treatment of fermented soybean oil and differentiation induction medium decreased $PPAR{\gamma}$ and $C/EBP{\alpha}$ gene expression at a concentration of $50{\mu}g/mL$ and blocked adipocyte differentiation by increasing adiponectin gene expression. The inhibitory effect of adipocyte differentiation was greatest in the BLO group. C57BL/6J mice were examined for 4 weeks after being separated into seven groups [normal diet group (N), high fat diet group (C), group fed high fat diet combined with regular soybean oil (SO), group fed non-fermented soybean oil (NF), and groups fed high fat diet combined with 5% fermented soybean oil (BA, BS, LBA, and BLO)] to identify the effects of soybean oil on body weight, serum lipid, adiponectin, insulin, and leptin levels in mice with high fat diet-induced obesity. The body weight and serum lipid level of the C group increased drastically compared to those of the N group. In contrast, the group fed a diet combined with fermented soybean oil showed decreases in weight, serum total cholesterol, LDL-cholesterol, and triglyceride levels compared to those of the C group. Moreover, soybean oil was found to be effective in the BLO group. In conclusion, fermented soybean oil has positive effects in prohibiting adipocyte differentiation increased by high fat diet and improving serum lipid composition. Therefore, fermented soybean oil can be used as a functional food material with anti-obesity activity.

Primary Culture of Chicken Tracheal Epithelial Cells and Study on Those Characters for Recombinant Virus Infection (닭 기도 상피세포의 1차 배양과 유전자 재조합 바이러스의 감염 특성 연구)

  • Park, Mi Na;Jang, Hyun-Jun;Keum, Dae Ho;Choi, Jin Ae;Yoo, Jae Gyu;Byun, Sung June;Park, Jong Ju;Ji, Ju Young;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Korean Journal of Poultry Science
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    • v.40 no.4
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    • pp.299-304
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    • 2013
  • Tracheal epithelial cells (TECs) are an important tool for studies of viral respiratory diseases. Primary TECs have been cultured from human, mouse and hamster. It is also necessary to diagnose viral respiratory disease and reveal infection mechanisms in chicken. In this study, we isolated tracheal epithelial layers from tracheal of 20-day-old chicks and cultured primary TECs from the isolated layers. Ciliated cells which were a typical morphology of TECs were observed in cultured primary TECs and maintained until cell passage 5 (15 to 20 days). When we analyzed expression patterns of epithelial marker genes (retinoic acid responder, FGF-binding protein, virus activating protease (VAP) in TECs compared to immortalized chicken embryonic fibroblast cell line (DF-1), all the marker genes are highly expressed in TECs than in DF-1. When TECs were cultured with 0.1 and 1 MOI of ND virus (rNDV-GFP strain) to test the susceptibility of TECs for ND virus, 12.6% and 48.2% of the incubated TECs were infected respectively. In addition, when DF-1 was incubated with 1 MOI of ND virus, the virus infection rate of DF-1 was three times lower than the virus infection rate of TECs. These data could contribute to study infection mechanisms of viral respiratory diseases and control them in chicken.

Antioxidant Effect of Enzymatic Hydrolysate from Sargassum thunbergii Using Vibrio crassostreae PKA 1002 Crude Enzyme (Vibrio crassostreae PKA 1002 유래 조효소액에 의한 지충이 (Sargassum thunbergii) 분해물의 항산화 효과)

  • Bark, Si-Woo;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Pak, Won-Min;Ahn, Na-Kyung;Choi, Yeon-Uk;Park, Ji-Hye;Bae, Nan-Young;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.105-111
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    • 2015
  • An alginate degrading enzyme from the Vibrio crassostreae PKA 1002 strain was used to hydrolyze the water extract of Sargassum thunbergii. To obtain the optimum degrading conditions for the S. thunbergii water extract, the mixture of the water extract and enzyme was incubated at 30℃ for 0, 3, 6, 12, and 24 h, and its alginate degrading ability was measured by reducing sugar and viscosity. A temperature of 30℃ for a period of 6 h was found to be the optimal condition for the enhancement of the alginate’s degrading ability. The pH of the enzymatic hydrolysate was not significantly different from that of the water extract. Overall lightness decreased, but redness and yellowness increased after enzymatic hydrolysis. Total phenolic compounds did not differ between the water extract and the enzymatic hydrolysate. DPPH radical scavenging activity and the reducing power of the enzymatic hydrolysate were lower than those of the water extract. However, the chelating effect of the enzymatic hydrolysate (80.08% at 5 mg/ml) was higher than that of the water extract (62.29%). These results indicate that the enzymatic hydrolysate possesses an anti-oxidant activity by way of the action of the chelating effect.

Plant Growth Promotion of Calystegia soldanella and Ischaemum anthephoroides by the Strain Penicillium citrinum KACC43900 (Penicillium citrinum KACC43900에 의한 갯메꽃과 갯쇠보리의 생장촉진활성)

  • You, Young-Hyun;Hwang, Jeong-Sook;Yoon, Hyeok-Jun;Khan, Sumera Afzal;Rim, Soon-Ok;Bae, Jeong-Jin;Choo, Yeon-Sik;Lee, In-Jung;Kong, Won-Sik;Lee, Byung-Moo;Kim, Jong-Guk
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1373-1377
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    • 2010
  • Coastal sand-dune plants can survive very effectively with the help of various microbes, especially ecto- and endomycorrihizae. Penicillium citrinum KACC43900 was screened according to growth promoting activity on sand-dune plants Calystegia soldanella and Ischaemum anthephoroides. In this study, coastal sand-dune plants were treated with a fungal culture filtrate of endophytic fungus P. citrinum KACC43900 to confirm the promotion of plant growth. C. soldanella and I. anthephoroides were used as representative coastal sand-dune plants, and their shoot length, plant length, and dry weight were analyzed. The shoot lengths of control C. soldanella and I. anthephoroides were 8.60 cm and 8.12 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 16.30 cm and 10.56 cm, respectively. The plant lengths of control C. soldanella and I. anthephoroides were 14.90 cm and 14.80 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 24.06 cm and 17.06 cm, respectively. The dry weight of C. soldanella and I. anthephoroides treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 0.163 g and 0.032 g, respectively. It was shown that the growth of shoots in C. soldanella and I. anthephoroides was promoted, 89.54% and 29.60%, by culture filtrate of P. citrinum KACC43900.

Effect of Interleukin-12 on the Expression of E-selectin in Mouse Model of Lewis Lung Carcinoma (Lewis 폐암 마우스 모델에서 Interleukin-12가 E-selectin 발현에 미치는 영향)

  • Lee, Sang-Haak;Shin, Yoon;Yoon, Hyoung-Kyu;Lee, Sook-Young;Kim, Seok-Chan;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.2
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    • pp.161-171
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    • 1999
  • Background: Interleukin-12 (IL-12) can induce antitumor effects in vivo. This antitumor effect is associated with T cell infiltration but the effect of IL-12 on the steps of T cell migration into the tumor tissue has not been fully elucidated. This study focused on the effect of IL-12 on the tumor growth and the metastasis and on the expression of E-selectin, an adhesion molecule which is activated endothelial specific in its expression. In addition, we studied whether the expression of E-selectin is associated with the TNF-$\alpha$, a cytokine that its production is increased by IL-12 and has functions inducing a variety of adhesion molecules. Methods: Mice of C57BL/6 strain were injected with Lewis lung cancer cells followed by either IL-12, TNF-$\alpha$, or normal saline by intraperitoneal route. Twenty eight days after tumor cell inoculation, metastatic nodules of lung were enumerated and immunohistochemical staining of the subcutaneous tumors were performed with monoclonal antibodies to CD4, CD8, CD16, and E-selectin. In IL-12 treated mice, the subcutaneously implanted Lewis lung tumors were decreased in size and the metastases were also decreased in number compared to control mice. On tumor tissues, increased infiltration of CD4+, CD8+, and CD16+ cells were oberved in IL-12 treated mice compared to control mice. In control mice, E-selectin was absent on tumor vessels, but the expression of E-selectin was increased on tumor vessels of IL-12 treated mice. Administration of TNF-$\alpha$ increased not only the expression of E-selectin but also infiltrations of CD4+, CD8+, and CD16+ cells on tumor tissues. Conclusions: These results demonstrate that IL-12 inhibits tumor growth and metastases through infiltrations of inflammatory cells in mouse model of Lewis lung carcinoma and E-selectin may playa role in inflammatory cell recruitment on tumor tissue following IL-12 administration. Also, TNF-$\alpha$ may have a role as a mediator responsible for the IL-12 induced expression of E-selectin.

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