Arbitrarily Primed PCR을 이용한 한국에서 유행하는 황색포도상구균의 분자유전형에 대한 연구

황선철,이창규,이승관,이동호,정수경,최현일,윤건석,정운원,윤효숙 高麗大學校 倂設 保健大學 保健科學硏究所 1998 保健科學論集 Vol.24 No.1

Eighty methicillin-resistant Staphylococcus aureus(MRSA) isolates were typed by applying arbitrarily primed-PCR(AP-PCR) method to clarify their distribution of molecular genetical characteristics. Among 40 gentamicin resistant strains of MRSA(GR-MRSA), 33 isolates drawn on the dendrogram fell into a single cluster at the similarity level of 90% when primer S₁ was used. However, with the primer S₂, 24 out of 40 strains fell into a single cluster at the similiarity 90%. In the meantime, 22 out of 40 strains amplified fell into a single cluster at the similarity of 90% when the primer E₂ was used. From the combined data obtained, it can be statistically said that 65.8% of GR-MRSA isolates are related with genetical characteristics. In 40 gentamycin susceptible MRSA(GS-MRSA) strains, 18, 19 and 13 strains drawn on the dendrogram fell into a single cluster at a similiarity level of 90% with the primers S₁, S₂ and E₂, respectively. From the combined data obtained by the three above AP-PCR profiles, it can be concluded that 41.7% of GS-MRSA isolates showed high relatedness genetically.

Multiplex PCR에 의한 methicillin내성 황색포도상구균의 분류

이승관,이창규,이동호,박상숙,윤효숙,황선철 高麗大學校 倂設 保健大學 保健科學硏究所 1996 保建科學硏究論集 Vol.5 No.1

In Staphytococcus aureus, mecA and nuc genes are the genetic determinants of methicillin resistance and thermostable nuclease. Using a multiplex PCR strategy. 533- and 267-bp regions of mecA and nuc genes, respectively, were coamplified to identify susceptible(lacking mecA) and resistant (mecA) Staphylococci and to differentiate S. aureus(nuc^(+)) from coagulase negative Staphylococci(lacking nuc). Two hundred seven Staphylococcal strains were tested. All 125 methidllin-resistant strains were found to be mecA^(+), and 33 of the 34 susceptible isolated lacked mecA. 37 methicillin resistant coagulase-negative Staphylococci(CNS) were also found to be mecA^(+), while 11 susceptible isolates lacked mecA. The nuc determinant was a unique feature of S. aureus ', it was found in 100% of the S. aureus strains tested but was not detected in all the coagulase-negative Staphylococci(CNS). In conclusion, coamplification of mecA and nuc determinants proved to be very reliable both for the rapid detection of methicillin resistance and differential diagnosis between S. aureus and other Staphylococci. This technique could be a useful tool not only in the diagnosis and classification of drug resistant pathogenic microorganisms but also for the monitoring the strategy of treatment for staphylococcal infections.

Ferritin H-like subunit from Manila clam (Ruditapes philippinarum): Molecular insights as a potent player in host antibacterial defense

Kim, H.,Elvitigala, D.A.S.,Lee, Y.,Lee, S.,Whang, I.,Lee, J. Academic Press 2012 Fish & Shellfish Immunology Vol.33 No.4

Ferritins are iron chelating proteins, which involve in iron metabolism and sequestration, contributing to the iron homeostasis in living organisms. In the present study, one ferritin subunit which was identified as H-like subunit was completely characterized in cDNA and protein levels from Manila clam (Ruditapes philippinarum); (RpFeH). The full length cDNA of RpFeH was 776 bp and it consisted of open reading frame of 513 bp, encoding a 171 amino acid peptide with a calculated molecular mass of 19.6 kDa and isoelectric point of 5.23. The amino acid sequence resembled the characteristic features of typical ferritin H subunits, including seven metal ligands in ferroxidase center, two iron binding region signatures and potential bio-mineralization residue (Thy<SUP>27</SUP>). Moreover, it was possible to identify an iron response element in 5' untranslated region, showing an agreement with previously reported ferritin H like subunits. The generated 3 dimensional tertiary structure of RpFeH showed a substantial consistency with the human ferritin H subunit, reinforcing its validity. Recombinant RpFeH was overexpressed in Escherichia coli BL21 (DE3) and purified. The recombinant protein showed a detectable iron binding activity according to the results obtained in our iron chelating activity assay. Furthermore it showed a noticeable anti-bacterial activity through suppressing the growth of Vibrio tapetis bacteria. The quantitative real time PCR detected ubiquitous RpFeH expression in tissues examined. However expression was found to be elevated in hemocyte and gill tissues. Transcriptional profile of Manila clam gill tissue, challenged with V. tapetis demonstrated a prolonged significant (P < 0.05) transcriptional up-regulations from 12 h post injection to 48 h post injection. Our findings suggest that RpFeH functions as an iron chelating protein subunit in Manila clam while contributing to the innate immune responses against bacterial infections, via its iron withholding function.

문학반응이론에 의한 보건의학기술계열 국가시험개선방안에 관한 연구

이창규,이승관,조경진,박종성,정수경,유병서,박상숙,윤효숙,황선철,문경환,김정민,함용운,김지환,임국환,김영순,윤경희,황성준 高麗大學校 倂設 保健大學 保健科學硏究所 1998 保健科學論集 Vol.24 No.1

The nationally-governed examinations for certification of allied health professions in Korea have been continued for thirty three years. During that time, there were a lot of managerial improvements in carrying out the examinations, for example, the looking-over the papers converted from manual method to computerized one. Nevertheless, the overall aspects of item management in the national examinations are still remained as obsolete style. In some developed countries they have already tried or adopted computerized system in making questions, executing item analysis, developing item banking, and in overall management of examinations, looking over papers for their national licensure examinations, and have established good reputations. Since the National Health Personnel Licensing Examination Board was established in 1998, now we can expect there would be a lots of improvements in the managerial systems and organizational structures associated with the national licensing examinations suggesting followings. 1. The contents and scope of the licensing examinations based on job analysis should be announced publicaly. 2. Items should be developed based on the scopes and contents of job characteristics. 3. All the developed items for the licensing examinations should be tested quantitatively prior to banking items. 4. All the parameters of the stored items should be fully estimated through item response theory. 5. All the efforts should be given in order to execute the examinations in individual residential areas on behalf of every examinee. 6. To the licensing examination system the Computerized Adaptive Testing system should be introduced in order to enhance the efficiencies. 7. Security enhancement on the stored items should be reminded in order to prevent leaking out the banking items. 8. Much more improvements should be made for the proper job evaluations. 9. Every efforts should be given to prevent cheatings encountable during the examinations. 10. Make the best of professional volunteers from the various fields in the execution of licensing examinations. 11. Consider a new system in that examinations can be executed twice or more in a year. 12. The current methods of presiding over the examinations should be replaced by a more reasonable one. 13. The results of the examinations should be announced as soon as possible in order not to prolong the examinees' unemployment periods. 14. The National Health Personnel Licensing Examination Board should try to rationalize the management keeping step with the information-oriented society.

Characterization and expression analysis of a goose-type lysozyme from the rock bream Oplegnathus fasciatus, and antimicrobial activity of its recombinant protein

Whang, I.,Lee, Y.,Lee, S.,Oh, M.J.,Jung, S.J.,Choi, C.Y.,Lee, W.S.,Kim, H.S.,Kim, S.J.,Lee, J. Academic Press 2011 Fish & shellfish immunology Vol.30 No.2

Lysozyme (muramidase) represents an important defense molecule of the fish innate immune system. Known for its bactericidal properties, lysozyme catalyzes the hydrolysis of β-(1,4)-glycosidic bonds between the N-acetyl glucosamine and N-acetyl muramic acid in the peptidoglycan layer of bacterial cell walls. In this study, the complete coding sequence of a g-type lysozyme (RBgLyz) was identified in the Oplegnathus fasciatus rock bream fish genome by means of multi-tissue normalized cDNA pyrosequencing using Roche 454 GS-FLX(TM) technology. RBgLyz is composed of 669 bp, with a 567 bp open reading frame that encodes 188 amino acids. Protein motif searches indicated that RBgLyz contains the soluble lytic transglycosylase domain involved in maintaining cell wall integrity. Furthermore, RBgLyz shares significant identity (81.4%) with Chinese perch Siniperca chuatsi. Quantitative real-time RT-PCR analysis results showed that RBgLyz transcripts are constitutively expressed in various tissues from healthy rock breams. In order to determine RBgLyz function in immunity, its expression was analyzed in head kidney following exposure to known immune stimulants or pathogens. RBgLyz transcripts were significantly up-regulated in response to challenge with lipopolysaccharide (LPS) and Edwardsiella tarda, as compared to non-injected control fish. Polyinosinic:polycytidylic acid (poly I:C) dsRNA stimulated a moderate expression of RBgLyz, as did Streptococcus iniae but to a lesser extent. There were no specific time-dependent effects on RBgLyz mRNA expression observed in response to rock bream iridovirus (RBIV) infection. Taken together, the gene expression results indicated that g-type lysozyme plays a role in the innate immune response to LPS, poly I:C, E. tarda and S. iniae in rock bream. Thus, we generated recombinant RBgLyz in an Escherichia coli expression system and characterized its antimicrobial activity. Our results indicated that recombinant RBgLyz had lytic activity against Gram-negative Vibrio salmonicida, Gram-positive Listeria monocytogenes, S. iniae and Micrococcus lysodeikticus. In addition, observations by scanning electron microscope (SEM) confirmed that the cell morphology of M. lysodeikticus was altered in the presence of recombinant RBgLyz.

Wilkie's Syndrome : 1例報告

金炯遠,睦榮珪,黃龍燮,柳建一,郭鎭榮 최신의학사 1971 最新醫學 Vol.14 No.12

The Wilkie's syndrome is an unusual form of duodenal obstruction due to compression of the distal duodenum between the aorta and the superior mesenteric artery. Authors report one case of duodenal obstruction due to superior mesenteric syndrome (Wilkie's syndrome) in 16-years old male student. A well development under nourished thin 16-years old male was admitted with abrupt onset of colicky abdominal pain and intermittent vomiting for two days, who no specific past, history except postprandial fullness. The diagnosis of Wilkie's syndrome was made under radiologic findings and above chief complaints. The operative treatment was performed by side to side anastomosis of- the jejunum to the dilated duodenum. We report a case of Wilkie's syndrome with brief review of several literature.

卵巢妊娠과 子宮內妊娠이 倂合된 一例

黃俊錫,李炅雨,朴勝昌,金永漢,金相浩 仁川基督病院 1977 仁川基督病院誌 Vol.1 No.1

Studies on the Detection of mecA Gene and Distribution of Enterotoxin Genes in Pathogenic Staphylococci Isolated from Korea Using Molecular Biologic Technique

Whang, S. C.,Lee, D. H.,Ryu, J. R.,Jung, S. K. 高麗大學校 倂設 保健大學 保健科學硏究所 1997 保建科學硏究論集 Vol.6 No.1

본 연구에서는 입상 검체에서 발견되는 병원성 포도상구균(S. aureus)과 methicillin 내성 포도상구균(MRSA)에서 각각 nurc과 mecA 유전자, 장독소인 enterotoxin A에서 enterotoxin E를 encoding하는 유전자들과 toxic shock syndrome toxin(tst) 유전자를 빠르개 탐지하고 독소형의 국내·외의 역학적인 분포를 조사하기 위하여 multiplex PCR을 수행하였다. 210개의 MRSA에서 두 개의 균주가 mecA 유전자 음성을 보인 것을 제외하고 모두 nuc유전자와 mecA유전자 양성을 나타냈다. 이 중 10개(80.5%)가 한 게 흑은 그 이상의 장독소에 대한 유전자를 지니고 있었으며 다른 41개 균주(19.5%)는 장독소 유전자에 대해 양성을 나타내지 않았다. 장독소A 유전자(sea)는 분리균 중 50.5%(n=105)에서 유전자가 중폭되었고 seb는 14.료쏭(n=30), sec는 23.3% (n=49), sed는 10.0%(n=21) 그리고 tst는 12.4%(n=26)가 증폭되었다. 58개의 균주들(27.68)은 두 개 이상의 장내독소 유전자가 증폭되었다. 가장 많이 나타난 독소 조합군은 sea와 seb로 26개 균주(12.4%)에서 발견되었으며 다음으로 빈번한 조합은 sec와 tst로 14균주(6.7%)로 나타났다. 210개 중 109개의 균주는 서울 지역에서 분리된 것이었고 101개는 부산 지역에서 분리된 균이었다. 부산지역에서 장내독소 A 유전자(sea)의 발생율은 서울에서보다 두 배 더 높게 나타났다(57.1% 대 23.6%). 그러나 장내독소 c유전자(sec)와 d유전자(sed)는 서울 지역에서 각각 23.6%와 17.9%로 비슷하게 나타난 반면에 부산 지역에서는 각각 2.0% 미만으로 거의 무시할 수 있을 정도로 낮게 나타났다. 한국의 장독소 형별 검출을 외국과 비교하면 장독소A의 검출빈도가 가장 높은 것은 같으나 차순으로 빈도가 높은 것이 외국의 경우 b유전자인데 반하여 한국에서는 c유전자로 실험결과 나타났다. 또한 한국에서의 d유전자 검출빈도가 b유전자 보다 높아 주된 유행형인 a유전자를 제외한 기타 독소형의 분포는 국가간 또는 지역간에도 차가 있는 것으로 결론지을 수 있다. In this study we attempted multiplex PCR for rapid detection of genes for nuc, mecA. enterotoxin A to E and TSST-l(tst) in bacterial lysates extracted from human strains of S aureus. In 210 isolates of methicillin-resistant Staphylococcus aureus (MRSA), all strains showed both nuc gene and mecA gene positive except two strains that showed only nuc gene positive. One hundred sixty nine(80.5%) of 210 MRSA isolates tested produced one or more toxins. Other 41(19.5%) isolates did not produce any of the toxins. Enterotoxin A gene(sea) was amplified in 50.5%(n=105) of the isolates, enterotoxin B(seb) in 14.3%(n=30), enterotoxin C(sec) in 23.3%(n=49), enterotoxin D(sed) in 10.0%(n=21) and TSST-l(tst) in 12.4%(n=26). Fifty eight strains(27.6%) amplified more than two enteroxin genes. The most common combination of toxin gene was sea and seb, which was found in 26(12.4%) strains, and the next frequent combination was sec and tst in 14(6.7%) strains from 210 isolates among which 109 MRSA strains tested were isolated in Seoul area and 101 in Pusan area. The frequency rate of enterotoxin A in Pusan was twice higher than that of Seoul with 57.1% to 23.6%. But the frequency rates of enterotoxins C and D genes in Seoul area are analogous with relatively narrow margin while those rates in Pusan area were almost negligible with less than 2% each. Apparent difference of enterotoxin type distribution was also found between the countries. Beside enterotoxin A which was most predominantely found in both European countries and Korea, enterotoxin C was the subsequently prevalent type of toxin in Korea instead of enterotoxin b which was found as the second most prevalent enterotoxin in some European countries

PCR(Polymerase Chain Reaction) 분석법을 이용한 Methicillin-Resistant S. aureus(MRSA)의 분류 실험

이승관(S. G, Lee),이창규(C. K. Lee),황선철(S. C. Whang),이도현(D. H. Lee),이동호(D. H. Lee) 고려대학교 보건과학연구소 1995 보건과학논집 Vol.4 No.1

Glycerol을 이용한 삼차신경통의 치료

박상근,정상섭,박형천,김용완,황용순,김기욱,주진양 대한신경외과학회 1985 Journal of Korean neurosurgical society Vol.14 No.3

Ten patients with trigeminal neuralgia were treated by the injection of glycerol into the trigeminal cistern. Eight patients became painless after the treatment, but one patient needed second injection for the relief of pain. Although follow-up period was less than one year these eight patients were free from pain without troublesome sensory changes. These preliminary results suggest that trigeminal neuralgia can be relieved by a simple percutaneous technique whose effect is not dependent on dense analgesia. But the long-term result of pain relief after the injection of glycerol remains to be determined.