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High Frequency of TP53 but not K-ras Gene Mutations in Bolivian Patients with Gallbladder Cancer

  • Asai, Takao;Loza, Ernesto;Roig, Guido Villa-Gomez;Ajioka, Yoichi;Tsuchiya, Yasuo;Yamamoto, Masaharu;Nakamura, Kazutoshi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5449-5454
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    • 2014
  • Although genetic characteristics are considered to be a factor influencing the geographic variation in the prevalence of gallbladder cancer (GBC), they have not been well studied in Bolivia, which has a high prevalence rate of GBC. The purpose of this study was to examine the frequency of TP53 and K-ras mutations in Bolivian patients with GBC and to compare them with our previous data obtained in other high-GBC-prevalence countries, namely Japan, Chile, and Hungary. DNA was extracted from cancer sites in paraffin-embedded tissue from 36 patients using a microdissection technique. TP53 mutations at exons 5 to 8 and K-ras mutations at codons 12, 13 and 61 were examined using direct sequencing techniques. The data obtained were compared with those in the other high-GBC-prevalence countries. Of the 36 patients, 18 (50.0%) had a TP53 mutation (one mutation in each of 17 patients and three mutations in one patient), and only one (2.8%) had a K-ras mutation. Of the 20 TP53 mutations, 12 were of the transition type (60.0%). This rate was significantly lower than that in Chile (12/12, P<0.05). In addition, three mutations were of the CpG transition type (15.0%), which is a feature of endogenous mutation. All three were found in the hot spot region of the TP53 gene. In contrast, G:C to T:A transversion was found in Bolivia, suggesting the presence of exogenous carcinogens. Our findings suggest that the development of GBC in Bolivia is associated with both exogenous carcinogens and endogenous mechanisms. The identification of an environmental risk factor for GBC is needed to confirm these findings.

Energy Metabolism and Methane Production in Faunated and Defaunated Sheep Fed Two Diets with Same Concentrate to Roughage Ratio (70:30) but Varying in Composition

  • Chandramoni, Chandramoni;Jadhao, S.B.;Tiwad, C.M.;Haque, N.;Murarilal, Murarilal;Khan, M.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.9
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    • pp.1238-1244
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    • 2001
  • Two calorimetric experiments were performed to investigate the effects of two diets with same concentrate: roughage ratio (70:30) but varying in composition on energy metabolism and methane production in faunated (F) and defaunated (DF) Muzaffarnagari sheep. For experiment I, ten animals were divided equally into two groups of which one was kept normally F as such while other was DF using 10% sodium lauryl sulphate. All the animals were offered diet I which comprised of oat hay and concentrate mixture I (CM I) containing maize grain (93%) as a major ingredient in 70:30 ratio. Similarly, the experiment II was conducted for which four F and four DF sheep (same as used for experiment I) were switched to diet II that consisted of maize hay and CM II (maize grain 59% + molasses 36%). Through diet II, DM intake in DF sheep was significantly (p<0.05) lower. Intake of GE through both the diets was similar in F and DF sheep. Digestibility of DM, OM, CP and GE and also metabolisability (ME/GE) was similar in F and DF sheep on both the diets. Total urinary energy loss did not differ in F and DF on both the diets, but methane energy loss as a percent of GE in DF was significantly (p<0.05) lower on diet I (3.75 vs 2.48), while it did not differ on diet II (3.20 vs 3.60). Heat production was significantly (p<0.01) reduced in DF on both the diets. Although, efficiency of utilisation of ME for maintenance calculated as per ARC (1984) did not differ in F and DF on both the diets, efficiency for maintenance and growth was higher (0.60 vs 0.672) on diet I in DF. It was inferred that methane production in DF sheep reduces on good quality hay-based diet supplemented with slowly fermentable carbohydrate (maize grain) but supplementation of molasses (rapidly fermentable CHO) nullify this effect when sheep were fed diets with concentrate: roughage ratio of 70:30.

Construction and Production of Concatameric Human TNF Receptor-Immunoglobulin Fusion Proteins

  • Yim, Su-Bin;Chung, Yong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.81-89
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    • 2004
  • Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

Effect of stocker management program on beef cattle skeletal muscle growth characteristics, satellite cell activity, and paracrine signaling impact on preadipocyte differentiation

  • Vaughn, Mathew A.;Lancaster, Phillip A.;Roden, Kelly C.;Sharman, Evin D.;Krehbiel, Clinton R.;Horn, Gerald W.;Starkey, Jessica D.
    • Journal of Animal Science and Technology
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    • v.61 no.5
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    • pp.260-271
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    • 2019
  • The objective of this study was to determine the effect of different stocker management programs on skeletal muscle development and growth characteristics, satellite cell (SC) activity in growing-finishing beef cattle as well as the effects of SC-conditioned media on preadipocyte gene expression and differentiation. Fall-weaned Angus steers (n = 76; $258{\pm}28kg$) were randomly assigned to 1 of 4 stocker production systems: 1) grazing dormant native range (NR) supplemented with a 40% CP cottonseed meal-based supplement ($1.02kg{\cdot}steer^{-1}{\cdot}d^{-1}$) followed by long-season summer grazing (CON, 0.46 kg/d); 2) grazing dormant NR supplemented with a ground corn and soybean meal-based supplement fed at 1% of BW followed by short-season summer grazing (CORN, 0.61 kg/d); 3) grazing winter wheat pasture (WP) at high stocking density (3.21 steers/ha) to achieve a moderate rate of gain (LGWP, 0.83 kg/d); and 4) grazing winter WP at low stocking density (0.99 steers/ha) to achieve a high rate of gain (HGWP, 1.29 kg/d). At the end of the stocker (intermediate harvest, IH) and finishing (final harvest, FH) phases, 4 steers / treatment were harvested and longissimus muscles (LM) sampled for cryohistological immunofluorescence analysis and SC culture assays. At IH, WP steers had greater LM fiber cross-sectional area than NR steers; however, at FH, the opposite was observed (p < 0.0001). At IH, CORN steers had the lowest Myf-5+:Pax7+ SC density (p = 0.020), while LGWP steers had the most Pax7+ SC (p = 0.043). At FH, CON steers had the highest LM capillary density (p = 0.003) and their cultured SC differentiated more readily than all other treatments (p = 0.017). At FH, Pax7 mRNA was more abundant in 14 d-old SC cultures from HGWP cattle (p = 0.03). Preadipocytes exposed to culture media from proliferating SC cultures from WP cattle isolated at FH had more $PPAR{\gamma}$ (p = 0.037) and less FABP4 (p = 0.030) mRNA expression compared with NR cattle. These data suggest that different stocker management strategies can impact skeletal muscle growth, SC function, and potentially impact marbling development in growing-finishing beef cattle.

SURFACE CHARACTERISTICS AND BIOACTIVITY OF ANODICALLY OXIDIZED TITANIUM SURFACES (양극산화에 의한 티타늄 산화막의 표면 특성 및 생체 활성에 관한 연구)

  • Lee, Sang-Han;Cho, In-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.1
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    • pp.85-97
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    • 2007
  • Statement of problem: Recently, anodic oxidation of cp-titanium is a popular method for treatment of titanium implant surfaces. It is a relatively easy process, and the thickness, structure, composition, and the microstructure of the oxide layer can be variably modified. Moreover the biological properties of the oxide layer can be controlled. Purpose: In this study, the roughness, microstructure, crystal structure of the variously treated groups (current, voltage, frequency, electrolyte, thermal treatment) were evaluated. And the specimens were soaked in simulated body fluid (SBF) to evaluate the effects of the surface characteristics and the oxide layers on the bioactivity of the specimens which were directly related to bone formation and integration. Materials and methods: Surface treatments consisted of either anodization or anodization followed thermal treatment. Specimens were divided into seven groups, depending on their anodizing treatment conditions: constant current mode (350V for group 2), constant voltage mode (155V for group 3), 60 Hz pulse series (230V for group 4, 300V for group 5), and 1000 Hz pulse series (400V for group 6, 460V for group 7). Non-treated native surfaces were used as controls (group 1). In addition, for the purpose of evaluating the effects of thermal treatment, each group was heat treated by elevating the temperature by $5^{\circ}C$ per minute until $600^{\circ}C$ for 1 hour, and then bench cured. Using scanning electron microscope (SEM), porous oxide layers were observed on treated surfaces. The crystal structures and phases of titania were identified by thin-film x-ray diffractmeter (TF-XRD). Atomic force microscope (AFM) was used for roughness measurement (Sa, Sq). To evaluate bioactivity of modified titanium surfaces, each group was soaked in SBF for 168 hours (1 week), and then changed surface characteristics were analyzed by SEM and TF-XRD. Results: On basis of our findings, we concluded the following results. 1. Most groups showed morphologically porous structures. Except group 2, all groups showed fine to coarse convex structures, and the groups with superior quantity of oxide products showed superior morphology. 2. As a result of combined anodization and thermal treatment, there were no effects on composition of crystalline structure. But, heat treatment influenced the quantity of formation of the oxide products (rutile / anatase). 3. Roughness decreased in the order of groups 7,5,2,3,6,4,1 and there was statistical difference between group 7 and the others (p<0.05), but group 7 did not show any bioactivity within a week. 4. In groups that implanted ions (Ca/P) on the oxide layer through current and voltage control, showed superior morphology, and oxide products, but did not express any bioactivity within a week. 5. In group 3, the oxide layer was uniformly organized with rutile, with almost no titanium peak. And there were abnormally more [101] orientations of rutile crystalline structure, and bonelike apatite formation could be seen around these crystalline structures. Conclusion: As a result of control of various factors in anodization (current, voltage, frequency, electrolytes, thermal treatment), the surface morphology, micro-porosity, the 2nd phase formation, crystalline structure, thickness of the oxide layer could be modified. And even more, the bioactivity of the specimens in vitro could be induced. Thus anodic oxidation can be considered as an excellent surface treatment method that will able to not only control the physical properties but enhance the biological characteristics of the oxide layer. Furthermore, it is recommended in near future animal research to prove these results.

An Approach to Determine the Good Seedling Quality of Grafted Tomatoes (Solanum Lycopersicum) Grown in Cylindrical Paper Pot Through the Relation Analysis between DQI and Short-Term Relative Growth Rate (DQI와 단기 상대생장률 분석을 이용한 원통형 종이포트 토마토 접목묘의 우량묘 기준 설정)

  • Seo, Tae Cheol;An, Se Woong;Jang, Hyun Woo;Nam, Chun Woo;Chun, Hee;Kim, Young chul;Kang, Tae Kyung;Lee, Sang Hee
    • Journal of Bio-Environment Control
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    • v.27 no.4
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    • pp.302-311
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    • 2018
  • Using cylindrical paper pot nursery method, three kinds of commercial tomatoes 'Dafnis', 'DOTAERANG DIA' and 'Maescala' were grafted onto a commercial rootstock 'B blocking'. From 10 to 40 days after graft-take, growth traits of seedlings were investigated by 0.5, 1.0 and 2.0S treatments of standard nutrient solution(S) for seedling growth, and top to root ratio(TRR), compactness(CP) and Dickson Quality Index(DQI) were calculated. Two weeks after transplanting of the seedlings under three different night temperature targeting to 10, 15, and $25^{\circ}C$, which were not precisely controlled, the relative growth rate (RGR) was investigated. The quantitative growth traits of grafted seedlings increased with increasing fertilizer concentration, and various range of seedling size could be produced. Compactness and DQI were significantly regressed (Adj $R^2=0.9480$). Short-term RGR after transplanting was higher at 1.0S treatment of standard nutrient solution at the seedling age of 30 days and 40 days after graft-take(DAGT). DQI and RGR were significantly regressed linearly at respective fertigation strength. Specially the diminishing slope of RGR was lower at 1.0S fertigation strength with the increase of DQI than others. The results indicate that DQI could be applied as a quality index of grafted tomato seedlings and the relation analysis between DQI and short-term RGR also could be used to determine the good quality seedlings of grafted tomato grown in cylindrical paper pot.

First Report of the Virus Diseases in Victory Onion (Allium victorialis var. platyphyllum) (산마늘(Allium victorialis var. platyphyllum)에서 바이러스병의 최초보고)

  • Park, Seok-Jin;Nam, Moon;Kim, Jeong-Seon;Lee, Yeong-Hoon;Lee, Jae-Bong;Kim, Min-Kyeong;Lee, Jun-Seong;Choi, Hong-Soo;Kim, Jeong-Soo;Moon, Jae-Sun;Kim, Hong-Gi;Lee, Su-Heon
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.66-74
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    • 2011
  • In 2005, a survey was conducted to identify virus diseases on victory onion, Allium victorialis var. platyphyllum grown in Ulleung island located in the East Sea. A total of 61 samples were collected from victory onion in the neighborhood of Seonginbong. The identification of viruses from the samples were carried out by electron microscopy and RT-PCR using primers species specific to GCLV, LYSV, SLV, OYDV and genus specific to Allexivirus, respectively. From sixty-one samples, filamentous rod particles (600-900 nm) were detected from four victory onion samples in EM, three samples containing SLV and one sample containing both SLV and Allexivirus in RT-PCR analysis, respectively. Victory onions naturally infected by the viruses were asymptomatic apparently. The viruses detected by RT-PCR were further characterized by the nucleotide sequence analysis of the coat protein region. Three isolates of SLV showed approximately 99% identities in the nucleotide and amino acid sequences, suggesting that they were likely to be the same strain. On the other hand, they showed approximately 75.7~83.7% identities in the nucleotide and 89.2~97.0% in amino acid sequences compared with the previously reported SLV isolates in Allium. The CP gene of the Allexivirus showed approximately 99.2% nucleotide identities and 98.8% amino acid identities with Garlic virus A. However, there was relatively low homology ranging from 60.6% to 81.5% compared with other Allexiviruses (GarV-C, GarV-E, GarV-X, GMbMV, and Shal-X). These data suggested that two viruses, SLV and GarV-A identified from victory onion, are named SLV-Ulleungdo and GarV-A-Ulleungdo, respectively. This is the first report of viruses infecting victory onion.

The Use of Alfalfa Hay for Starter Diet of the Korean Native Goats (한국재래산양의 이유 후 초기 사료로 알팔파 건초의 이용)

  • 조익환;황보순;전하준;안종호;이주삼;한태호
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.2
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    • pp.115-122
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    • 2002
  • In this study, the effects of alfalfa hay included in different ratios of 60%(A), 50%(B), 40%(C) and 30%(D) for the diets of young weaned Korean native goats were investigated on feed intake, digestibilities of nutrients, nitrogen retention, average daily gain and feed efficiency. Crude protein contents of experimental diets were 18% in all treatments. The contents of ADF, NDF and crude ash were higher in A as 26.1, 37.0 and 9.0% compared to the lowest values in D of 20.3, 30.4 and 6.6%. However the contents of non-structural carbohydrates was in the opposite tendency. Daily feed intake (DM) was highest in A (414.4g) and B (417.7g) and significantly (p<0.05) lower in D(362.6g). Dry matter intake per metabolic basal weight (DM g/kg of $BW^{0.75}$ ) were highest in A(73.9g) and lowest in D(64.0g). Average daily gain and feed efficiencies were in the range of 33.3~48.7g and 8.1~13.4% respectively with no statistical differences, however with the increasing trends according to higher percent of roughage in the diets. Digestibilities of dry matter (DM), organic matter (OM), crude protein (CP), ADF, NBF, crude ash (CA) and non-structural carbohydrates (NSC) were in the tendency of becoming higher according to higher ratios of concentrate but becoming lower according to higher ratio of alfalfa hay in the diets fed to Korean native goats. Nitrogen retention was lowest in A (1.8g, 15.5%) and highest in D (2.7g, 25.7%). In overall, inclusion of 30~40% alfalfa hay as a roughage in the diets of weaned Korean native goats was superior to the higher rates of inclusion of alfalfa hay in aspects of average daily gain and feed efficiency. The fixed 18% of crude protein content in the diets was efficient for the growth of weaned goats.

Effect of Dietary Betaine and Energy Levels on Liver Fats and Cholesterol in Laying Hens (사료내 비태인과 에너지 수준이 산란계의 간지방과 콜레스테롤에 미치는 영향)

  • Park J. H.;Park S. Y.;Ryu K. S.
    • Korean Journal of Poultry Science
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    • v.32 no.3
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    • pp.149-156
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    • 2005
  • Two experiments were conducted to examine the effects of betaine intake on blood and yolk cholesterol, abdominal fat, liver fat, tissue triglyceride(TG) and liver HMG-CoA reductase In laying hens. In Expt. 1, a total of 72 ISA-brown laying hens were individually assigned into four treatments from 18 to 21 weeks old. Com-soybean meal based diet were fed with the addition of 0, 300, 600 and 1,200ppm. In Expt. 2, 72 ISA-brown laying hens were housed into individual cage to evaluate the effect of dietary betaine(0, 600ppm) and energy(ME, 2,800, 2,900kca/kg) from 70 to 74 weeks. Serum total cholesterol, LDL and HDL-cholesterol and TG concentration in blood of hens fed betaine tended to increase compared to those of the control, but were not significantly different. However, betaine supplementation showed a statistically significant decrease in yolk cholesterol(P<0.05). There was no significant difference in abdominal fat among the treatments. Liver fats and 7c of birds 130 betaine was decreased compared with control. Serum total cholesterol, LDL-cholesterol and triglyceride concentration were significantly inc.eased by ffeding a diet containing 600ppm betaine in Expt. 2(P<0.05), but were not influenced by the dietary energy levels. Yolk cholesterol, abdominal fat and HMG-CoA reductase activity were affected neither by dietary energy nor betaine level.

Influence of Supplemental Dietary Yeast Culture on the Noxious Gas Emission in Broiler Houses and Performance of Broiler Chicks (효모배양물 첨가 사료가 계사내 유해가스 발생 및 육계의 생산성에 미치는 영향)

  • Park, J. H.;Ryu, M. S.;Kim, S. H.;Na, C. S.;Kim, J. S.;Ryu, K. S.
    • Journal of Animal Science and Technology
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    • v.45 no.1
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    • pp.41-48
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    • 2003
  • Two experiments were conducted to investigate the effect of dietary supplementation of yeast culture on the performance of broiler chicks and noxious gas emission in broiler houses. Two hundred forty and three hundred and twenty, one day old Cobb male broiler chicks in Expt 1 and Expt 2, respectively were alloted to four treatment levels of yeast culture (0, 0.1, 0.2, 0.4%). To each treatment, 60 birds were assigned in Expt 1 and 5 replicates of 16 birds each were assigned in Expt 2. Basal diets contained 21.5% and 19.0% CP, and 3,100kcal/kg ME for the starting and finishing periods, respectively. Ammonia and $CO_2$ gas emission were detected twice a day for seven days during the five week period of Expt 1. Weight gain, feed intake and feed efficiency were measured for five weeks in Expt 2. Intestinal microbes, blood cholesterol and ND antibody titer were examined at the end of Expt 2. In Expt 1, the concentration of $NH_3$ in the house of birds fed yeast culture tended to be lower than the control. It was significantly lower in the 0.4% yeast culture treatment than the control (P<0.05). $CO_2$ concentration was significantly lower in all yeast culture treatments regardless of its dietary supplemental level than the control (P<0.05). different from others. Feed efficiency (feed/gain), however, was significantly improved in all yeast culture treatments relative to that of the control for starting period (P<0.05) and that of 0.2% yeast culture treatment was significantly lower than those of the control and 0.4% for the overall period. Total number of E. coli in the ileum of birds fed yeast culture at 0.1 and 0.2% was significantly lower than those of the control and 0.4% in the ileum. The CFU of Lactobacillus spp. of birds fed yeast culture at 0.1% was higher in the cecum compared to other treatments (P<0.05). Total cholesterol level of chicks fed 0.1% yeast culture seemed to be lower compared to that of other treatments, whereas LDL-cholesterol level was significantly lower than those of the control and 0.4% treatment. ND antibody titer tended to be higher in the yeast culture treatments than the control, but was not significantly different. The results of these experiments indicated that 0.2% yeast culture may have a potential to reduce the noxious gas emission in broiler houses and maximize the performance of broiler chicks.