• Korean Journal of Microbiology
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• v.46 no.1
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• pp.21-26
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• 2010

As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

• Journal of Applied Biological Chemistry
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• v.53 no.1
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• pp.13-20
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• 2010

Biological activities such as anti-oxidative and anti-microbial of the Seungmakalgeuntang, a traditional prescription, were evaluated. The electron donating ability of water, ethanol, supercritical fluid and 1,3-butylene glycol extract of Seungmakalgeuntang showed more than 50% at a 100 ppm concentration. At a 1000 ppm concentration, the superoxide dismutase-like activities of ethanol and supercritical fluid extract of Seungmakalgeuntang showed less than 50%. xanthine oxidase inhibition effect of the supercritical fluid extract showed more than 70% at a 1,000 ppm concentration, which was higher than vitamin C. From the measurement on lipid oxidation, the $Fe^{2+}$ chelating abilities of the supercritical fluid extract of Seungmakalgeuntang was more than 60% at a 100 ppm concentration. Also the $Cu^{2+}$ chelating abilities of supercritical fluid extract Seungmkalgeuntang was showed more than 60% at a 500 ppm concentration. Clear zones formed by sample against the human skin-resident microflora such as Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acne of ethanol and supercritical fluid extract of Seungmakalgeuntang showed the highest among all the extracts tested using a 4mg/disc. The minimum inhibitory concentration (MIC) against both S. epidermidis and S. aureus showed 2,500 ppm in the extract of the supercritical fluid.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.12
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• pp.1745-1752
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• 2010

This study was conducted to investigate the antimicrobial activity of 70% ethanol (EtOH) extract and the five fractions of the crude extract from Sasa borealis leaves against seven food poisoning bacteria, Staphylococcus aureus, Micrococus luteus, Listeria monocytogens, Bacillus subtilis, Escherichia coli, Salmonella Typhimurium, and Pseudomonas aeruginosa. The yield of 70% EtOH extract was 11.4% and those of n-hexane, chloroform, ethyl acetate, n-butanol, and aqueous fractions were 3.0%, 1.1%, 0.6%, 1.3%, and 5.1%, respectively. The 70% EtOH extract and the four fractions except aqueous fraction demonstrated antimicrobial activity against all the seven food poisoning bacteria at a concentration of 0.5%, although it was less compared to benzoic acid. Minimal inhibitory concentration (MIC) of the 70% EtOH extract against all the food poisoning bacteria except S. aureus was $50{\mu}L$/disc. Moreover, chloroform fraction was $35{\mu}L$/disc against 3 food poisoning bacteria and $50{\mu}L$/disc against the other 4 food poisoning bacteria; ethyl acetate fraction was $50{\mu}L$/disc against all the food poisoning bacteria. In addition, n-butanol fraction was $50{\mu}L$/disc against all the food poisoning bacteria except S. aureus. Aqueous fraction, which did not show antimicrobial activity at 5%, was $200{\mu}L$/disc against only S. aureus and L. monocytogen. The 0.25%, and 0.5% of ethyl acetate fraction inhibited the growth of all the food poisoning bacteria 8 to 12 hours and 24 hours, respectively. These results indicate that the Sasa borealis leaves may be useful as a natural antimicrobial substance.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.12
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• pp.1867-1872
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• 2010

Microbe and quality changes of vacuum-packaged ready-to-eat lettuce were analyzed. While the vacuumpackaged lettuce after chlorine sanitizer were stored at $5^{\circ}C$, $15^{\circ}C$, and $25^{\circ}C$ for 7 days, viable numbers of total aerobic bacteria (TAB), coliform, E. coli, food-borne pathogens and lactic acids bacteria (LAB) were counted with gas production and sensory evaluation. Before the storage, only TAB of 2 log CFU/g and coliform of 1 log CFU/g were detected and LAB was not detected. TAB, coliform and LAB increased by 1 log CFU/g at $5^{\circ}C$ for 7 days without any detection of the pathogens. Sensory evaluations for off-flavour and crispness dropped to half the best value at 5 day storage. TAB and coliform increased by 3 log CFU/g and 2 log CFU/g, respectively, but LAB grew very actively by 4 log CFU/g under anaerobic environment and only B. cereus were detected after enrichment of the lettuce at $15^{\circ}C$ for 3 days. The evaluations for off-flavour and crispness were half the best value for 3 days. However, TAB and coliform increased by 3 log CFU/g, 1 log CFU/g, and 4 log CFU/g, respectively only at 1 day storage under $25^{\circ}C$. Also B. cereus were detected after enrichment and the sensory evaluation were half the best value within 1 day storage. Therefore, preservation at the lowest temperature possible is required for growth inhibition of the bacteria contaminated in the lettuce. Interestingly, LAB among them grew most actively under the anaerobic condition and the adulteration of lettuce might be closely related with the growth of LAB.

• The KIPS Transactions:PartA
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• v.11A no.7 s.91
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• pp.555-562
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• 2004

Both global volume reduction and local shape changes of hippocampus within the brain indicate their abnormal neurological states. Hippocampal shape analysis consists of two main steps. First, construct a hippocampal shape representation model ; second, compute a shape similarity from this representation. This paper proposes a novel method for the analysis of hippocampal shape using integrated Octree-based representation, containing meshes, voxels, and skeletons. First of all, we create multi-level meshes by applying the Marching Cube algorithm to the hippocampal region segmented from MR images. This model is converted to intermediate binary voxel representation. And we extract the 3D skeleton from these voxels using the slice-based skeletonization method. Then, in order to acquire multiresolutional shape representation, we store hierarchically the meshes, voxels, skeletons comprised in nodes of the Octree, and we extract the sample meshes using the ray-tracing based mesh sampling technique. Finally, as a similarity measure between the shapes, we compute $L_2$ Norm and Hausdorff distance for each sam-pled mesh pair by shooting the rays fired from the extracted skeleton. As we use a mouse picking interface for analyzing a local shape inter-actively, we provide an interaction and multiresolution based analysis for the local shape changes. In this paper, our experiment shows that our approach is robust to the rotation and the scale, especially effective to discriminate the changes between local shapes of hippocampus and more-over to increase the speed of analysis without degrading accuracy by using a hierarchical level-of-detail approach.

• Korean Journal of Poultry Science
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• v.39 no.1
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• pp.63-70
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• 2012

This study was conducted to determine the possible use of Red Ginseng marc as stress inhibiter in thermal stress (temperature humidity index 86) and lipopolysaccharide (LPS) - exposed laying hens by investigating their effects on laying performance, blood biochemical parameters, immunoglobulin concentration and serum superoxide dismutase (SOD) like ability. A total of forty-five 52-wk-old laying hens (ISA Brown) were divided into 3 treatment groups with 5 replicates of 3 birds in each group. NC (negative control, no immune substances), PC (positive control, ${\beta}$-glucan 25 ppm) and RGM (Red Ginseng Marc 3%) were added in feed with respective substance. Egg production in RGM was significantly increased in comparison with NC groups for 8 weeks (P<0.05). On blood biochemical parameters, effects of ambient temperature is definite by showing significant difference in aspartate aminotransferase and others (P<0.05), but RGM both before and after thermal stimulation have no significant difference in comparison with other groups. And for 3 weeks after thermal stimulation, laying performance was also not significantly different among treatments. Immunoglobulin M content and SOD like activities after challenge with LPS were higher in the RGM and PC than NC (P<0.05). In conclusion, although ineffective as inhibiter in thermal stress, dietary supplementation of Red Ginseng marc improved SOD like activity and immune system by regulating immunoglobulin content in laying hens. These findings have laid the foundation for future studies of immunomodulation in laying hens fed Red Ginseng Marc and of evaluation of heat stress inhibitor.

• Korean Journal of Poultry Science
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• v.35 no.3
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• pp.303-311
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• 2008

This study was conducted to investigate the effect of dietary supplementation of Cu-methionine chelate(Cu-Met) and Cu-soy proteinate(Cu-SP) on the performance, small intestinal microflora and immune response in laying hens. A total of 960 Hy-line $Brown^{(R)}$ laying hens of 39 wks old were assigned to one of the following 6 dietary treatment: control(C), antibiotic(Avilamycine 6 ppm), Cu-Met 50 and Cu-Met 100(50 and 100 ppm Cu as Cu-methionine chelate), Cu-SP 50 and Cu-SP 100(50 and 100ppm Cu as Cu-soy proteinate). Each treatment was replicated 4 times with forty birds per replication, housed in 2 birds per cages. Forty birds units were arranged according to randomized block design. Feeding trial lasted 6 wks under 16 hours lighting regimen. Hen-day and hen-house egg production of groups treated with Antibiotic and Cu supplements tended to be higher than the control with significant difference (P<0.05) shown between Cu-Me 100 and control. Egg weight was significantly (P<0.05) heavier in antibiotic and Cu-SP treatments than Cu-Met treatments but they were not significantly different from the control. Eggshell strength, egg shell thickness, egg yolk color and Haugh unit were not significantly different among treatments. There were no significant differences in leukocytes and erythrocytes in the chicken blood. But mean corpuscular hemoglobin value(MCH) was significantly (P<0.05) higher in Cu-SP 100 than antibiotic treatment. The concentrations of serum IgG and IgA were not significantly different among treatments. Copper concentration in the liver tended to increase as the level of copper supplementation increased, that of Cu-SP 100 being significantly (P<0.05) higher than those of the control and antibiotic treatment. Concentrations of iron and zinc of the liver were not significantly influenced by treatments. Populations of Cl. perfringens and Lactobacilli in the small intestinal content were significantly (P<0.05) influenced by treatments. Population of Cl. perfringens decreased and that of Lactobacilli increased in the copper supplemented groups. The result of this experiment showed that Cu-Met and Cu-SP are comparable to antibiotic in improving egg production in laying hens. Birds fed diets supplemented with Cu-SP produced heavier eggs than those fed diets with Cu-Met. There were no significant differences in the performances between 50 ppm and 100 ppm copper supplementation as organic forms.

• Korean Journal of Poultry Science
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• v.41 no.3
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• pp.165-171
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• 2014

The objective of this experiment was to investigate the effect of dietary supplementation of copper-soy proteinate (Cu-SP) and herbal mixture (HBM) on growth performance, intestinal microflora, and immune response in broiler. A total 1,000 1-d old ROSS 308 (initial $BW=41{\pm}0.38g$) were randomly allotted to 1 of 5 dietary treatments with 4 replicates during d 35 of the feeding trial. Dietary included: (1) Control : control diet, (2) Antibiotics : control diet + Avilamycin 6 ppm, (3) Cu-SP : control diet + 100 ppm Cu-soy proteinate, (4) HBM : control diet + 0.15% herbal mixture, (5) Cu-SP+HBM : control diet + 100 ppm Cu-soy proteinate + 0.2% herbal mixture. Two-phase feeding program with a starter diet from d 0 to 21, and a finisher diet from d 22 to 35 was used in the experiment. Within each phase, a diet was formulated to meet or exceed NRC requirements of broilers for macro- and micronutrients. The diet and water were available ad libitum. Result indicated that during d 22 to 35 and over all periods of the experiment, feed intake and feed conversion ratio were greater (P<0.05) for other treatment than control. Significant differences were found in lymphocyte (LY), and stress indicator (HE:LY ratio). Lymphocyte was greater (P<0.05) for Cu-SP + HBM treatment than control. However, stress indicator (HE:LY ratio) were greater (P<0.05) for control than Cu-SP + HBM treatment. The plasma IgG was higher (P<0.05) in the antibiotics, HBM, and Cu-SP+HBM treatments groups compared with control. The population of Clostridium perfringens in the antibiotics, Cu-SP, HBM, Cu-SP + HBM treatment groups were lower (P<0.05) than those control. These result suggested that dietary copper-soy proteinate or herbal mixture may be used as an alternative to antibiotics to improve growth performance, and intestinal health of birds.

• Korean Journal of Poultry Science
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• v.40 no.4
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• pp.283-290
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• 2013

This experiment was conducted to investigate the effects of dietary bacteriophage CP supplementation on growth performance, nutrient digestibility, blood profiles, visceral organ weight, meat quality and fecal microflora in broilers. A total of 340 1-d-old ROSS 308 broilers (mixed gender) with an initial average body weight (BW) of $41.14{\pm}0.17g$ were randomly allotted to 4 treatments with 5 replicate pens per treatment and 17 broilers per pen for 31 days. Dietary treatments were: 1) CON, control diet, 2) CP05, CON + 0.05% bacteriophage CP, 3) CP10, CON + 0.10% bacteriophage CP and 4) CP15, CON + 0.15% bacteriophage CP. During d 15 to d 31, broilers fed CP15 diet had higher (P<0.05) body weight gain and feed intake than broilers fed CON diet. Overall, body weight gain in CP10 and CP15 treatment groups was greater (P<0.05) than that in CON treatment and feed intake was higher (P<0.05) in CP15 treatment than that in CON. Apparent total tract nutrient digestibility and blood characteristics did not differ (P>0.05) among treatments. The water holding capacity of breast meat increased (P<0.05) in broiler fed the diets containing bacteriophage CP compared with those fed the CON diet. Other meat characteristics such as pH value, breast muscle color ($L^*$, $a^*$, $b^*$) and drip loss were unaffected by dietary supplementation of bacteriophage CP. The weight of bursa of Fabricius increased (P<0.05) in CP05 when compared with CON. No significant difference was observed (P>0.05) among treatments in visceral weight and fecal microflora concentrations of Lactobacillus spp., Clostridium perfringens, Escherichia coli and Salmonella spp. In conclusion, dietary supplementation with 0.10 and 0.15% bacteriophage CP could improve the growth performance.

• BMB Reports
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• v.38 no.1
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• pp.115-119
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• 2005

Replacement of valine by tryptophan or tyrosine at position $\alpha$96 of the $\alpha$ chain ($\alpha$96Val), located in the ${\alpha}_1{\beta}_2$ subunit interface of hemoglobin leads to low oxygen affinity hemoglobin, and has been suggested to be due to the extra stability introduced by an aromatic amino acid at the $\alpha$96 position. The characteristic of aromatic amino acid substitution at the $\alpha$96 of hemoglobin has been further investigated by producing double mutant r Hb ($\alpha$42Tyr$\rightarrow$ Phe, $\alpha$96Val$\rightarrow$Trp). r Hb ($\alpha$42Tyr$\rightarrow$Phe) is known to exhibit almost no cooperativity in binding oxygen, and possesses high oxygen affinity due to the disruption of the hydrogen bond between $\alpha$42Tyr and $\beta$99Asp in the ${\alpha}_1{\beta}_2$ subunit interface of deoxy Hb A. The second mutation, $\alpha$96Val$\rightarrow$Trp, may compensate the functional defects of r Hb ($\alpha$42Tyr$\rightarrow$Phe), if the stability due to the introduction of trypophan at the $\alpha$96 position is strong enough to overcome the defect of r Hb ($\alpha$42Tyr$\rightarrow$Phe). Double mutant r Hb ($\alpha$42Tyr$\rightarrow$Phe, $\alpha$96Val$\rightarrow$Trp) exhibited almost no cooperativity in binding oxygen and possessed high oxygen affinity, similarly to that of r Hb ($\alpha$42Tyr$\rightarrow$Phe). $^1$H NMR spectroscopic data of r Hb ($\alpha$42Tyr$\rightarrow$Phe, $\alpha$96Val$\rightarrow$Trp) also showed a very unstable deoxy-quaternary structure. The present investigation has demonstrated that the presence of the crucible hydrogen bond between $\alpha$42Tyr and $\beta$99Asp is essential for the novel oxygen binding properties of deoxy Hb ($\alpha$96Val$\rightarrow$Trp).