• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1053-1058
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• 2003

The solvent extracts of Houttuynia cordata root, which were extracted by using several solvents with different polarities, were prepared for utility as a natural preservatives. The antimicrobial activity was investigated by disc diffusion method against 22 microorganisms consisting of food borne pathogens, food poisioning microorganisms and food-related bacteria. The extraction yields were 15.7%, 3.7%, 0.13%, 0.5% and 5.9% in ethanol, chloroform ethylacetate, butanol and aqueous fractions, respectively. Antimicrobial activities were shown in ethanol, ethylacetate and butanol fraction of Houttuynia cordata root. However chloroform and aqueous fractions showed weak antimicrobial activity against the tested bacteria. Among the four fractions, ethylacetate fraction showed the strongest antimicrobial activities against microorganisms tested, such as B. megaterium, E. coli, K. pneumoniae and S. typhimurium. The polyphenolic compounds widely occuring in the traditional medicine plants have been reported to possess high antimicrobial activity. The polyphenolic compound in ethylacetate and butanol fraction were 35.9% and 16.0%, ethanol, chloroform and aqueous fraction were 5.0%, 2.3% and 1.7%, respectively. There are some relationship between antimicrobial activity and polyphenol content in natural plants. The ethylacetate fraction could be suitable for the development of a food preservative.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.3
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• pp.394-400
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• 2004

To investigate the efficacy of alternatives to antibiotics, the present study was conducted to compare the effects of antibiotic, lactic acid, a blend of commercial essential oils (EOs) and EOs in combination with lactic acid on growth performance and the functional activity of the gut in broiler chickens. A total of 168 broiler chickens were given the basal diet supplemented with 10 ppm colistin (T1), 0.1% lactic acid (T2), 25 ppm EOs (T3), 25 ppm EOs+0.1% lactic acid (T4), 50 ppm EOs (T5) or 50 ppm EOs+0.1% lactic acid (T6) in the period 3 to 35 days of age. As a result, the broiler chickens assigned to T4 group throughout the experimental period had apparently (p<0.05) greater body weight and total gain than these assigned to T1, T2, T3 and T5 groups. However, there was no difference in growth performance among the birds fed the diets supplemented with antibiotic (T1), lactic acid (T2) and EOs (T3 and T5) alone. The weights of digestive organs and the number of lactobacilli and E. coli in the lower ileum were not affected by dietary treatments. Total trypsin activity was significantly (p<0.05) greater in T4 than T1, T2, T3 and T5 groups. Total and specific pancreatic $\alpha$-amylase activities were significantly (p<0.05) enhanced in the broiler chickens fed T4 diet compared with these fed T1, T2 and T3 diets. However, there were no differences in growth performance and digestive enzyme activities including pancreatic trypsin and $\alpha$-amylase between T4 and T6 groups fed the diets supplemented with either low or high EOs levels in combination of lactic acid. In conclusion, a blend of commercial EOs combined with lactic acid showed significant increases in digestive enzyme activities of the pancreas and intestinal mucosa, leading to increase in growth performance.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.818-825
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• 2013

We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1749-1759
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• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

• Journal of Animal Science and Technology
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• v.59 no.8
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• pp.18.1-18.8
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• 2017

Background: In 2006, the European Union (EU) has decided to forbid use of antibiotics as growth promoters. Although many researches had been conducted about fiber source as alternatives of antibiotics, there are still lack of reports in the literature about the optimum level of sugar beet pulp supplementation, affecting growth performance and nutrient digestibility in weaning pigs. Therefore, different level of sugar beet pulp was added to diets to determine the effects of sugar beet pulp supplementation on growth performance, nutrient digestibility, fecal microflora, blood profile and incidence of diarrhea in weaning pigs. Methods: A total of 200 weaning pigs [$(Yorkshire{\times}Landrace){\times}Duroc$], averaging $9.01{\pm}1.389kg$ of initial body weight were, allotted to 5 treatments in a randomized complete block (RCB) design. Each treatment was composed of 4 replicates with 10 pigs per pen. The treatments were control treatment: Corn-SBM basal diet + ZnO (phase 1: 0.05%; phase 2; 0.03%) and four different levels of sugar beet pulp were supplemented in Corn-SBM basal diet (3, 6, 9 or 12%). Two phase feeding programs (phase 1: 1-2 weeks; phase 2: 3-5 weeks) were used for 5 week of growth trial. Results: In feeding trial, there were no significant differences in growth performance and incidence of diarrhea among treatments. The E.coli counts were not significantly different among dietary treatments but linear response was observed in Lactobacillus counts as sugar beet pulp supplementation increased (P < 0.05). In addition, IGF-1, IgA and IgG were not affected by dietary treatments. However, the BUN concentration was decreased when pigs were fed the treatments of diets with SBP compared to that of control treatment (P < 0.05). In nutrient digestibility, crude fiber and NDF digestibilities were improved as the sugar beet pulp increased (P < 0.05). However, digestibilities of crude ash, crude fat, crude fiber and nitrogen retention were not affected by dietary sugar beet pulp levels. Conclusion: This experiment demonstrated that sugar beet pulp can be supplemented in weaning pigs' diet instead of ZnO to prevent postweaning diarrhea without any detrimental effect on growth performance.

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.196-203
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• 2006

Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

• Microbiology and Biotechnology Letters
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• v.32 no.4
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• pp.297-306
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• 2004

Heat shock proteins (HSPs) were induced in cells in the thermal stress, and the HSP90 family is one of the major classes of HSPs. Gene encoding HSPs have been characterized from various mammals and piscine. We have cloned and sequenced the HSP90 cDNA from a brain cDNA library constructed from flounder (Paralichthys oliThe result of sequence analysis shows it to be the HSP90~. The nucleotide sequence of the HSP90$\beta$ was composed of 2791 long, encoding 726 amino acid residues. The flounder hsp90$\beta$ gene showed very high sequence homology with hsp90f3 of European sea bass (96.6%), zebrafish (92.9%), Atlantic salmon (92.0%) and human (89.5%). We also constructed a phylogenetic tree based on HSP90 amino acid sequences from vertebrate species. Gene-specific primers were selected and used in RT-PCR reactions to measure the basal hsp90$\beta$ mRNA. The hsp90f3 gene is constitutively expressed at a fairly high level in all examined tissues (brain, liver, kidney, muscle, and spleen). In order to express protein of flounder hsp90$\beta$ in E. coli, we used the His-tagged pETvector. Then, the expression of flounder HSP90$\beta$ was confirmed by Western blot analysis.

• Journal of Chest Surgery
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• v.21 no.2
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• pp.316-325
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• 1988

Author made a clinical study of 248 cases of pleural effusion patients who were diagnosed and treated at departments of chest surgery and internal medicine, Pusan National University Hospital, during the period from Jan. 1983 to Dec. 1985. The age distribution ranged from 1 to 76 years old and the ratio of male to female was 1.38:1. The cardinal symptoms were chest pain[69.4%], dyspnea[66.1%], cough[57.7%], fever[37.1%], sputum[26.2%], general malais[13.7%] and cyanosis[1.6%] in this order. The causes of pleural effusion were pulmonary tuberculosis[42.4%], pneumonia[23.0%], malignancy[16.5%], congestive heart failure[9.3%], liver cirrhosis[2.8%] and nephrosis[2.0%] in this order. The protein in the pleural effusions was 1.61*0.90[mean*SD] gm% in transudate and 5.05*1.10[Mean*SD] gm% in exudate. In 34 cases[89.5%]out of 38 transudates, the protein was under 3 gm% and in 201 cases [95.7%] out of 210 exudates, the protein was over 3 gm%. The protein ratio of pleural effusion to serum was 0.2650.11[Mean LSD] in transudates and 0.73*0.12[Mean LSD] in exudate. The ratio under 0.5 was in 36 cases[94.8%] out of 38 transudates and over 0.5 was in 206 cases[98.1%] out of 210 exudates. The LDH in the pleural effusion was 114.7550.3[mean*SD] units / ml in transudate and 627.05325.9[mean*SD] units / ml in exudate. The LDH less than 200 units / ml was in 36 cases[94.6%] out of 38 transudates and more than 200 units / ml was in 199 cases[94.7%] out of 210 exudates. The LDH ratio of pleural effusion to serum was 0.34k 0.11[mean*SD] in transudate and 1.15*1.12[mean*SD] in exudate. The LDH ratio of pleural effusion to serum was less than 0.6 in 36 cases[94.8%]out of 38 transudates and more than 0.6 in 200 cases[95.2%] out of 210 exudates. Etiologic organisms were confirmed in 78 cases[48.1%] among the requested 162 cases. In the 78 cases of etiologic organisms, staphylococcus was 33 cases[20.3%], streptococcus 24 cases[14.8%], Klebsiella pneumonia 7 cases[4.3%], pseudomonas 6 cases[3.7%], E. coli[3.1%], enterobacter 3 cases[1.9%]. 43 patient of pleural effusion from malignancy were undergone three or more thoracenteses. In 13 cases[31.7%], three specimen were negative and in 7 cases[17.1%], three specimens were positive for malignancy. In the remaining of 21 cases[51.2%], malignant cells were found in one or more of the specimens but not in all. Methods of treatment of pleural effusion by closed thoracotomy was 188 cases[75.8%], thoracentesis 27 cases[10.9%], decortication 16 cases[6.5%], thoracoplasty 6 cases[2.4%] and decortication with thoracoplasty 3 cases[1.2%].

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.665-680
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• 1996

Bovine Pneumonic Pasteurellosis는 수송열(輸送熱)로 일반적으로 알려져 있는 질병으로서, 여러가지 요인의 복합적(複合的)인 작용에 의해 발병하는 것으로 알려져 있으나, Pasteurella haemolytica A1이 가장 주요(主要)한 인자(因子)로 밝혀져 있다. P haemolytica A1은 leukotoxin(LKT), lipopolysaccharide(LPS), capsular polysaccharide 등 여러가지의 병원성인자(病原性因子)을 생성한다. 이들 인자중 LKT가 가장 중요한 병원성인자로 밝혀져 있다. 이에 본 실험은 P haemolytical A1의 LKT 유전자를 Bacillus subtilis에서 발현(發現)시킴으로서 LPS에 오염(汚染)되지 않은 LKT을 대량으로 생산할 목적으로 실시되었다. 실험의 첫 단계(段階)로서 pLKT52 plasmid을 Sau3 A1의 제한효소을 이용하여 부분소화(部分消化)시킨 후 이 부분 소화(消化)된 유전자들로부터 3~5kb 크기의 유전자들을 순수분리하여 pUC18와 결합시킨 후 E coli NM522에 형질전환(形質轉換)시켰다. 이때 형질전환된 균주들은 LKT에 대한 단크론 항체인 MAb601을 이용하여 colony blot 법에 의해서 LKT 유전자 보유 및 발현여부(發現與否)을 조사하였다. 이들 양성 clone들은 제한효소분석(制限酵素分析), 염기서열분석(鹽基序列分析) 및 Western blot 등에 의해서 재확인(再確認)하였다. 총 9개의 양성 clone중 위의 방법에 의해서 한 clone을 선택(選擇)하여 lktCA insert를 재분리하여 shuttle vector에 subcloning 하였다. Subcloning된 LKT 유전자들은 shuttle vector의 종류(種類)(pHPS9, p602/20, pHPS9-Sac)와 각기(各其) 다른 종류(種類)의 B subtilis(spoO12A, BR121, WB3O, Raj1105) 숙주내(宿主內)에서 발현정도를 Western blot 법에 의해서 비교(比較)하였다. 이때 최적발현조건(最適發現條件)은 p602/20와 pBL1의 dual plasmid system을 이용하여 B subtilis spoO12A에서 2시간동안 IPTG로 발현을 유도(誘導)하는 것이었다. B subtilis에서 발현된 LKT을 visual 법과 neutral red uptake 법을 이용하여 소 폐포(肺胞) 대식구(大食求)에 대한 biological activity를 확인하였다. 발현된 LKT에 대한 LPS 오염은 LKT을 SDS-PAGE 후 silver stain에 의해서 확인하였다. 본 실험을 통해서 볼 때에 lktCA 유전자를 보유(保有)하고 있는 p602/20는 B subtilis에서 매우 불안정(不安定)하였고, 발현된 LKT는 세균자체(細菌自體)에서 생성되는 protease들에 의해서 파괴(破壞)됨으로서 농도(濃度)가 매우 낮았다. 이러한 문제점들은 다음 단계(段階)의 실험에서 해결되어야할 문제들이다.

• The Korean Journal of Nuclear Medicine
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• v.23 no.2
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• pp.219-223
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• 1989

Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.