김치에서 분리한 Lactobacillus sp. JC-7과 Lactobacillus acidophilus 88간의 Electrofusion 최적조건 설정

조영배,최현정,백형석,전홍기 부산대학교 유전공학연구소 1997 분자생물학 연구보 Vol.13 No.-

김치 발효 숙성기간을 연장하고 신선도를 오랫동안 유지 할 수 있는 김치발효 starter의 개발을 목적으로 최적숙성기에 있는 김치로부터 bacteriocin생성능이 없는 유산균을 분리하여 동정하였으며, bacteriocin생성능이 우수한 L.acidophilus 88을 융합시키기 위해 electrofusion에 대한 최적 조건을 검토하였다. 분리균주는 Lactobacillus속으로 동정되어 Lactobacillus sp. JC-7이라 명명하였다. Electrofusion에 의해 생성된 융합주를 식별하기 위해 streptomycin(2.5㎎/ml)에 내성을 나타내는 Lactobacillus sp. JC-7변이주와 kanamycin(600㎍/ml)에 내성을 나타내는 L.acidophilus 88변이주를 분리하였다. Electrofusion을 100V/㎝, 120msec(72ohms,1670 capacitance) 에서 수행했을 때 융합효율이 가장 양호하였으며 전기장의 세기와 시간이 중가할수록 융합 효율이 현저히 감소하였다. 2가 양이온은 농도가 중가할수록 대체적으로 융합효율을 감소시키는 경향을 나타내었으나 1mM MGCl_2에서는 대조군에 비해 융합효율이 약간 증가하였다. PEG매개에 의한 융합법의 융합효율을 비교한 결과, 융합효율은 chemical fusion<electrofusion<electrofusion+20% PEG순을 나타내었다. Evaluation of Optimum Conditions for the Electrofusion between Lactobacillus sp. JC-7 Isolated from Kimchi and Lactobacillus acidophilus 88. Young-Bae Jo, Hyun-Jung Choi, Hyung-Suk Baik and Hong-Ki Jun*. Department of Microbiology, Pusan National University, Pusan 609-735,Korea-A lactic acid bacterium was isolated from kimchi. The isolated strain was identified as the genus Lactobacillus thrugh its morphological characteristics and named as Lactobacillus sp. JC-7. The optimum conditions for the electrofusion between streptomycin(2.5㎎/ml)resistant mutant of Lactobacillus acidophilus 88 and kanamycin(600㎍/ml) resistant mutant Lactobacillus sp. JC-7 were evaluated. The highest number of fusants were obtained at a capacitance value of 120msec(1670㎌), a field strength of 100V/㎝,and a pulse controller setting of 72Ω. The potimum pH of elecroporation buffer was 7.5 and the concentration of divalent cation was 1mM MG^2+. Electrofusants were efficiently obtained by addition 20% polyethylene glycol to electroporation buffer. The yield of fusion was better than that of using polyethylene glycol mediated chemical induction.

김치 분리균인 Bacillus sp. JK-43이 생산하는 Cyclodextrin Glucanotransferase의 생산 및 특성

전홍기,배경미,김영희,백형석 부산대학교 유전공학연구소 2000 분자생물학 연구보 Vol.16 No.-

김치 시료로부터 자동산화되지 않으며 열 및 중성 pH에서 안정한 AA 유도체인 AA-2G를 생산할 수 있는 당전이활성을 가진 CGTase 생산균주를 분리하였고, 분리균주의 형태학적, 배양학적, 생리학적 성질 및 16s-rDNA sequences를 조사한 결과 그람 양성의 간균으로 호기성이며 내생포자를 형성하는 전형적인 중온성 Bacillus sp. JK-43으로 동정되었다. Bacillus sp. JK-43의 CGTase는 AA-2G 뿐만 아니라 AA-6G로 추정되는 물질을 함께 생산하였으며, 효소 최적생산조건은 1.0% soluble starch, 1.0% yeast extract, 1.0% Na_2CO_3, 0.1% K_2HPO_4 그리고 0.02% MgSO_4·7H_2O가 함유된 배지에서 pH 7.0, 37℃에서 26시간 동안 진탕배양하였을 때였다. 각종 당공여체에 따른 Bacillus sp. JK-43의 AA-2G 생산성을 조사한 결과 β-CD에서 가장 높은 AA-2G 생산성을 보였으며, 식혜제조폐액인 엿기름 및 밥당화액에서도 비교적 높은 AA-2G 생산성을 보였다. 또한 여러 가지 당수용체에 대한 JK-43의 CGTase의 당전이 반응을 검토한 결과 sucrose, mannitol 및 inositol에서 높은 당전이 수율인 70∼90%를 나타내었다. A bacterial strain, designated as JK-43, producing extracellular cyclodextrin hlucanotransferase (CGTase)[EC 2.4.1.19] was isolated from kimchi. The CGTase from isolated strain JK-43 showed the transglucosylation activity from soluble starch to L-ascorbic acid (AA) compared to those obtained from other strains. A main product formed by this reaction was identified as 2-O-α-glucopyranosyl L-ascorbic acid (AA-2G) by testing its susceptibility to α-glucosidase hydrolysis, the HPLC prodiles, and through the elementary analysis. The β-CD, γ-CD, potato starch and corn starch were identified to be suitable glucosyl donor for transglucosylation reaction on AA by CGTase. Acceptor specificity on AA-2G production was examined by use of AA, Iso-AA and AA-2P. Transglucosylation was observed toward AA-2P as well as AA. The microorganism isolated from kimchi was identified as a strain of Bacillus sp. JK-43 based on the maximal CGTase production was observed in a medium containing 1.0% soluble starch, 1.0% yeast extract, 1.0% Na_2CO_3, 0.1% K_2HPO_4, and 0.02% MgSO_4·7H_2O with initial pH 7.0. The strain was cultured at 37℃ for 26 hrs with reciprocal shaking.

활성탄을 이용한 휘발성 유기화합물 (VOC) 제거

廉聖培,安埈炯 홍익대학교 산업기술연구소 2003 産業技術 Vol.13 No.-

In accordance with the development of modern industries, the amount of harmful volatile organic compound(voc) in exhaust gas has increased rapidly. VOC is one of main cause of air polution, so its removal is very important problem for public hygiene. In this study we try to remove of specific polution gas(toluene gas) in automobile exhaust gas by acrive carbon. The conclusion of this study as follow. 1. The amount of gas adsorbed by carbon is increased by falling temperature but for the inhibition of corrosion of the apparatus by HCl or SO2 operation must be carried above the dew point. 2. The amount of adsorbed gas is more at the condition of flow rate 0.1M/S but the unit amount of absorbed gas is more at te flow rate 0.2 M/S. 3. Total amount of absorbed gas increases in accordance with the thickness of active carbon bed but the unit amount of absorbed gas decrease reversely.

HBx형질전환 생쥐에서 발생한 간세포암종에서 H-ras 및 c-myc의 발현에 관한 연구

문형배,소병준,김학철,윤기중,한원철,조향정,유대열,정영진 圓光大學校 醫科學硏究所 2002 圓光醫科學 Vol.17 No.2

<연구목적> HBx형질전환 생쥐에서 발생한 간세포암종의 발암과정에 종양유전자(H-rgs, c-myc)의 발현 정도를 조사하고자 하였다. <연구방법> 정상생쥐 12마리(4-18개월) 및 HBx 형질전환 생쥐 44마리(4-18개월)를 대상으로 포르말린에 고정하고 파라핀에 포매한 간 조직을 이용하여 면역조직화학적염색을 실시하였다. 실험군은 정상 부위, 이형성 부위 및 종양 부위로 구분하였으며, 종양 부위는 소결절성병변 부위와 간세 포암종 부위로 구분하였고, 이형성병변 부위는 이형성병변만 발견되는 부위, 소결절성병변과 동반된 이형성병변 부위 및 간세포암종과 동반된 이형성병변 부위로 구분하였다. <연구결과> H-rgs의 발현은 정상 간조직에 비하여 이형성병변 부위(P<0.05) 및 종양 부위(P<0.01)에서 증가하였으며, 소결절성병변 부위과 간세포암종 부위 사이에서는 간세포암종 부위에서 증가된 경향이었으나 통계학적으로 유의한 차이가 없었으며, 각 이형성병변 부위 사이에서도 유의한 차이는 없었다. c-myc의 발현은 정상 간조직 및 이형성병변 부위에 비해 종양 부위에서 증가하였으며(P<0.001), 소결절성병변 부위와 간세포암종 부위에서는 비슷하였고, 각 이형성병변부위 사이에서도 비슷하였다. <결론> HBx형질전환 생쥐에서 발생하는 간세포암종의 발생에 H-rgs는 이형성 변화를 일으키는 시기에 관여하며, c-myc은 이형성병변에서 암으로 이행하는 시기에 관여할 것으로 생각한다. Background: This experiment was designed for the expression of H-ras and c-myc in hepatocarcinogenesis of the HBx transgenic mice. Methods: Immunohistochemical stains in the paraffin embedded tissue of the liver were used for the detection of H-ras and c-myc in the 12 normal mice and 44 HBx transgenic mice of the 4-18 month old. Results: Expression of the H-ras was significantly increased in the dysplastic area (P<0.05) and tumor area (P<0.01) than in the normal liver. But there were no differences of H-ras expression between areas of microscopically identified hepatocellular carcinoma (MI-HCC) and grossly identified hepatocellular carcinoma (GI-HCC) and dysplastic areas among the only dysplastic areas, dysplastic areas with MI-HCC and GI-HCC. Expression of the c-myc was significantly increased in the tumor area (P<0.001) than in the normal liver and dysplastic area. But there were no differences of c-myc expression between areas of MI-HCC and GI-HCC, and dysplastic areas among only dysplastic areas, dysplastic areas with MI-HCC and GI-HCC. Conclusions: Our study suggests that H-ras is related to the dysplastic change and c-myc is related to the neoplastic change in the hepatocarcinogenesis of the HBx transgenic mice.

랫드에서 인체 재조합 적혈구 조혈인자, rHuEPO의 13주 정맥투여 아만성독성에 관한 연구

김형식,곽승준,천선아,박현선,한하수,임소영,안미영,김원배,김병문,안병옥,홍성렬,이병무 성균관대학교 약학연구소 1998 成均藥硏論文集 Vol.10 No.1

A recombinant human erythropoietin (rHuEPO) was administered intravenously at dosage levels of 0, 100, 500, and 2500 IU/㎏/day for a period of 13 weeks. There were no observed clinical signs and deaths related to treatment in all groups tested. Decreases in body weight gain and food consumption were observed only in males of 2,500 IU/㎏ group after 2 weeks. In hematological parameters, erythrocyte content, hematocrit values and hemoglobin concentration were dose-dependently increased in rHuEPO treated groups. The ratio between kidney weight and whole body weight was significantly increased in females of 500 and 2,500 IU/㎏ groups. The spleen weight was also increased in both sexes of 500 and 2,500 IU/㎏ groups. However, the absolute weight change of other organs was not observed. In histopathological examinations, the renal tubular basophilia was observed only in males and females of 2,500 IU/㎏ groups. From these results, it is concluded that the no-observed adverse effect level(NOAEL) of rHuEPO is 100 IU/㎏ in rats in the present study.

흉골 골절의 의의에 관한 임상적 연구

원형섭,황성연,박정배,이준형,박철완,이근 대한응급의학회 1996 대한응급의학회지 Vol.7 No.2

It has been known that sternal fracture increases the risk of potentially life-threatening injuries, such as myocardial, pulmonary, and thoracic vascular injuries, and it has 25% to 45% of mortality, primarily resulting from these underlying injuries. The authors reviewed retrospectively the medical records and radiographic findings of the patients who admitted with sternal fracture at Chung Ang Gil Hospital from 1 June 1994 to 31 December 1995 to evaluate the significance and clinical course of sternal fracture. The results were followings: 1. The male to female ratio was 3:2 and the mean age was 43.1 and 45.4 years old in male and female, respectively. 2. Motor vehicle occupant accident was the most common cause of sternal fracture. 3. Of 33 cases, there was no death, but common associated injuries which were rib fracture, hemothorax, lung contusion, vertebral fracture, penumothorax, long bone fracture, myocardial contusion, flail chest, facial chest, facial bone fracture, and pericardial effusion, in order. 4. The clinical course of sternal fracture was determined primarily by associated injuries except for communited sternal fracture. 5. The patients with isolated sternal fracture, who had an otherwise normal chest x-ray, normal electrocardiogram, no other significant associated injuries and are hemodynamically stable, had a benign clinical course.

Lactobacillus acidophilus 88과 Lactobacillus bulgaricus IFO 13953간의 세포융합주의 특성에 관한 연구

조영배,김혜정,김성구,백형석,전홍기 부산대학교 유전공학연구소 1996 분자생물학 연구보 Vol.12 No.-

유산균의 균주개량방법의 일환으로 protoplast fusion 기법과 electrofusion법을 이용하여 protease 활성, lipase 활성, 내열성, 내산성 등이 우수한 L. bulgaricus 와 bacteriocin을 생산하는 L. acidophilus를 융합시켜 얻은 융합주들의 생리학적 성질을 검토하였다. 산 생성능, 내열성, 내산성, protease, lipase 활성 등은 L. bulgaricus가 L. acidophilus 보다 우수하였다. L. bulgaricus는 lactose와 sorbose를 이용하였으나 maltose와 sorbitol을 이용하지 못하는 반면, L. acidophilus는 maltose를 이용하고 lactose와 sorbose를 이용하지 못하였다. 융합주 가운데서는 3, 6, 7, 8, 10번이 모균주의 발효능 특성을 함께 지님으로서 재조합체임을 확인 할 수 있었으며, sorbitol의 경우 모균주에서는 발효능이 전혀 나타나지 않았음에도 불구하고 융합주 4, 7번이 발효능을 나타내어 융합과정에서 새로운 형질을 획득하기도 한다는 사실을 알 수 있었다. Lactase 활성은 모균주 모두 높은 β-galactosidase 활성을 보였으나, phospho-β-galactosidase 활성은 거의 없었으며 융합주들도 다소 차이는 있었지만 모균주와 유사한 효소 활성을 나타내었다. 발효에 있어서 key enzyme으로 작용하는 protease, lipase 등의 효소 활성도 모균주의 활성 보다 우수한 융합주도 존재하였다. Aninterspecific fusant was made from the protoplasts of two strains of lactobacillus genus (streptomycin and lincomycin resistant L. bulgaricus and kanamycin resistant L. acidophilus 88). The functional properties of the fusant were examined by determining bacteriocin productivity, acid producing activity, ability of carbohydrates fermentation and three important enzyme activities. The recombinant strain revealed bacteriocin productivity. Acid production and β galactosidase, phospho-β-galactosidase, lipase and protease activity of L. bulgaricus were better than those of L. acidophilus 88. Among fusants, β-galactosidase activity of two strains were better than that of the parent strains but phospho-β-galactosidase activity remarkably lower. One fusant revealed the improved proteolysis compared to that of the parent strains- Lipase activity of L. bulgaricus was better than that of L. acidophilus but another fusant exhibited the highest lipase activity.

TRISS Method와 ASCOT Method를 이용한 외상환자의 생존율 분석

김형수,배성만,양혁준,박철완,이근,고영관 대한응급의학회 1995 대한응급의학회지 Vol.6 No.2

Despite traumatic injuries cause serious problems in our society, there are few adequate and objective scoring system that assess the severtiy of trauma patients. The Major Trauma Outcome Study(MTOS)in United States deisgned the TRISS(Trauma Score & Injury Severity Score)method by means of Revised Trauma Score(RTS),Injury Severity Score(ISS0,and age,while Champion et al deviced ASCOT(A Severtriy Characterizaiton Of Trauma)method to overcome the limitation of TRISS. This study attempted to make a comparison between TRISS and ASCOT by using date for 422 injured patients which were collected form September,1993 to February,1994. Ascot and TRISS were compared in their sensutivity,Specificity,disparity and Z-statistics. 1)Sex ratio of male to female was 2.8:1 and the commonest age of trauma patients was thirties (23.8%). 2)The average probability of survival(Ps)for 442 patient by TRISS method was 0.9228 and that of ASCOT method 0.9356. 3)Disparity of Ps between survival and non-survival using TRISS and ASCOT was relatively low for both indexes as 0.3507 and 0.3296, respectively. 4)The sensitivity rates(number of patients predicted to die who actually died/total who actually died)for the non-survival of both TRISS and ASCOT method were low (35.0%),but the specificity rates(number of patients predicted to live who actually lived/total who actually lived)for the survival of TRISS and ASCOT were 99.1%and 99.6%respectively. 5)Z-statistics(difference between predicted and actual number of death)of both TRISS(1.3224)and ASCOT(1.2234)resulted in positive value which meant that actual number of death exeeded predicted number of death. 6)The ASCOT that have presumed to be more accurate method for patients with head trauma and with multiple injuries to one portion of body,had its intricacy and difficult points in practical application. And the difference between the result of ASCOT and TRISS was not so significant. 7)It is thought that a new, more comprehensive index would like to be developed and thoroughly tested on a variety of data sets in order for it to be used in trauma system quality assurance evaluation.

병아리에서 닭와포자충에 의한 F-낭염이 SRBC에 대한 항체형성에 미치는 영향

박배근,사또시, 가바모도,전무형,김무강,류시윤 충남대학교 수의과대학 동물의과학연구소 1996 動物醫科學硏究誌 Vol.4 No.-

병아리에 닭와포자충(Cryptosporidium baileyi)의 감염에 의한 F-낭염이 면양 적혈구(SRBS)에 대한 항체 형성에 미치는 영향을 관찰하기 위하여 2일령의 병아리에 닭와포자충의 오오시스트 1 × 10^6개를 경구 접종하였다. SRBC를 경정맥 및 F-낭의 관강에 감작하고 항체형성능과 F-낭의 조직소견을 1 주일 간격으로 10주간 관찰하여 다음과 같은 결과를 얻었다. 1. 체중의 증가율은 실험 전 기간을 통하여 전반적으로 실험군이 대조군 보다 낮았다. 2. F-낭 지수(Bursa index)는 1주령 부터 5주령 까지는 1이하로 위축되었으며, 6주령 부터 1이상으로 종창되었다. 특히, 6주령의 경우 1.94로 종창의 정도가 가장 심하였다. 3. 상피세포에서 발육하고 있는 닭와포자충의 오오시스트 및 점막하직과 여포내의 호산구의 침윤은 1주령에서 4주령 까지 관찰되었으며 1주령에 가장 많은 오오시스트가 상피에서 관찰되었다. 4. 상피세포 및 FAE의 핵융해, 비만세포의 출현, 여포의 괴사 및 상피의 증생은 5주령 이후에 관찰되었다. 상피세포의 핵융해는 6주령에 가장 심하였다. 5. HA titer는 정맥내로 항원을 감작한 군에서 7주령에 최고의 역가를 보였으며 대조군이 64±35, 실험군이 44.8±15.7이였다. F-낭의 관강내로 항원을 감작한 군에서는 8주령에 대조군이 11.2±3.9, 실험군이 2.8±1.0로 실험군의 역가가 모두 낮았다. 이상의 결과로 볼 때 닭와포자충의 감염에 의한 F-낭염(bursitis)은 SRBC에 대한 항체형성능을 저해The 함을 알 수 있다.

마우스에서 오존수의 급여가 쥐와포자충 감염에 미치는 영향

박배근,김덕환,류시윤,김현수,조성환,전무형 충남대학교 수의과대학 동물의과학연구소 2000 動物醫科學硏究誌 Vol.8 No.-

Ozone is very effective in acute and chronic viral disease as a virucidal agent in reversing positive antibody tests and in improving the general health of the animals. The effects of ozonized water supply against Cryptosporidium muris was studied in 5-weeks-old mice. After oral inoculation of mice with 1×10^6 oocysts, ozonized water(0.5ppm) and tap water were supplied to experimental and control group, respectively. The prepatent period of both group was 8 days postinoculation(PI). The patent period of experimental group was longer(62days) than that of experimental group(57days). The number of oocysts in the feces were maxium on days 28 PI, indicating 3.54×10^6 oocysts/experimental mouse and 5.56×10^6 oocysts/ control mouse. The number of discharge oocysts during 12-33 PI was more in control group than that of experimental group. In the view of gastric gland section on days 28 PI, the number of endogenous development of C muris was smaller in experimental group than in control group. This results indicates that the autoinfected sporozoite were damaged by ozone.