경남지역에서 분리한 Salmonella Enteritidis의 항생제 감수성 검사 및 random amplification polymorphic DNA (RAPD)-PCR을 이용한 유전형 분석

김은경 ( Eun-gyeong Kim ),김민경 ( Min-kyung Kim ),권현애 ( Hyun-ae Kwon ),윤도경 ( Do-kyung Youn ),구정헌 ( Jeong-heon Koo ),박소연 ( So-yeon Park ),이희근 ( Hui-geun Lee ),조명희 ( Myeong-hui Jo ),하도윤 ( Do-yun Hah ),김철호 ( 한국동물위생학회(구 한국가축위생학회) 2018 韓國家畜衛生學會誌 Vol.41 No.3

Salmonella Enteritidis (S. Enteritidis) are found in animals, humans, and environment. In addition, S. Enteritidis draws attention to the public health concerns due to carriage of antibiotic resistance traits. For these reasons, the prevalence and antibiotic resistance patterns of S. Enteritidis are significant issues with regard to public health. To address this issues, a total of 24 strains of S. Enteritidis from 164 samples collected from several slaughterhouses in Gyeong-Nam province in order for antibiotic resistance profiles. Subsequently, we characterized the genotyping by random amplification polymorphic DNA (RAPD)-PCR. As a result, very high level of resistance to protein synthesis inhibition antibiotics and most isolates were susceptible to others. Six random primers were used for RAPD-PCR to reveal genotypes of S. Enteritidis isolates. One of the primer, P1245, generated 147 distinct RAPD-PCR fragments ranging from 400∼3000 bp. The number of RAPD-PCR products ranged from 4 to 8 for this primer. The RAPD-PCR fragments could be placed these strains into 3 subgroups and 2 classes by UPGMA cluster analysis. Interestingly, several S. Enteritidis that isolated from different slaughterhouses showed same genotype. These results showed only limited genetic variation among the isolates, those were grouped into a few different patterns of antibiotic resistance.

위암세포에 의한 종양침윤 림프구의 면역반응 억제기전에 관한 연구

박정규,송규상,서광선,최정목,배진선,장일성,윤완희,노승무,조은경,백태현 大韓免疫學會 1995 大韓免疫學會誌 Vol.17 No.3

Tumor-infiltrating lymphocytes ('1°ILs) interact most closely with tumor cells and thus are more likely to reflect tumor host interactions accurately. But it is unknown whether such T cells are nonspecific inflammatory cells or a subset of specific host immune responses. In this study, there was no clear correlation between the infiltration of T lymphocytes in stomach cancer and the overexpression of c-ErbB-2 or increasing class I MHC expression on tumor cells. A positive correlation was seen between the presence of TILs in the tumor and tumors with diploidy by flow cytometric DNA analysis. The proliferative responses of Ills stimulated with IL-2, anti-CD3 mAb, or both were examined. When compared to normal mucosal-associated lymphoid tissue lymphocytes, the proliferative response of TILs to high dose IL-2 was minimal. A similarly poor response to anti-CD3 mAb plus IL-2 was also observed. The freshly isolated TILs exhibit reduced ability to proliferate in response to IL-2, anti-CD3 mAb or both. The microenvironment of the tumor suppresses the proliferative capacity of the TILs. The mechanism of this suppression remains unknown. It could be mediated by suppressor cells, by soluble substances within the tumor, or both. To examine this question, supernatants of stomach cancer cells (SNSNU-1) were tested for the presence of immunosuppressive factors. Human peripheral blood T-cells and tumor-draining lymph node lymphocytes (TDLNL) were incubated for 3 days with SNSNU-1 and then assessed for proliferative responses to PMA, anti-CD28 mAb, or both and for the inducibility to express IFN- r or IL-4 mRNA to PMA. Peripheral blood T-cells pretreated with SNSNU-1 were unable to proliferate in response to PMA, anti-CD28 mAb or both. SNSNU-1 also produces inhibitory activities of TDLNL proliferative response to PMA or anti-CD28 mAb and PMA (49%, 52%, respectively). In contrast, culture supernatants obtained from HEp-2, K562 or Daudi showed normal proliferative responsiveness of peripheral blood T-cells and TDLNL by PMA, anti-CD28 mAb or both.

양극성 장애에서 표피성장인자의 유전적 다형성에 대한 연합연구

김수경,이규영,주은정,강웅구,안용민,김용식 大韓神經精神醫學會 2007 신경정신의학 Vol.46 No.6

Objectives : Epidermal growth factor (EGF) is a neurotrophic factor which regulates the intracellular signaling molecules. These molecules are also affected by mood stabilizers such as lithium and valproate. In addition, epidermal growth factor enhances neuronal survival, maturation and differentiation especially in midbrain dopaminergic neurons of which dysfimction may play a role in pathophysiology of bipolar disorders. Bipolar disorder has some genetic commonalities with schizophrenia, and several association studies of EGF have been done with schizophrenia. In this study, we tried to investigate the geneticassociation between EGF A61G polymorphism and bipolar disorder. Methods : Total of 189 patients and 347 nonnal control were included. All patients satisfied the diagnostic criteria of DSM-IV for bipolar disorder type I (BPDI, N= 146) and bipolar disorder type II (BPDII, N=43). Genomic DNA was extracted from the peripheral blood, and genotyping was performed by TaqMan™ method. Genotype and allele frequency of EGF A61G polymorphism between the patients and the control were compared by contingency chi-square test or the Fisher's exact test. Results : No association was found between EGF A61G polymorphism and susceptibility of BPDI, BPDII and bipolar disorder (all patients). Female patients with BPDII showed overexpression of AG genotype compared to that ofcontrol group (p=0.03). However, this association was not significant after correction of multiple testing. Conclusion : In conclusion, EGF A61G polymorphism has no association with susceptibility of bipolar disorder. However, the disease modifying role of EGF gene polymorphism for bipolar disorder remains to be elucidated in respect to factors such as gender difference or diagnostic subtype.

Interleukin-2와 결핵균 30 kDa 항원이 구개편도 및 말초혈액 T 세포 증식에 미치는 상승효과

박정규,박찬권,조은경,김화중,백태현,고필준,김병국,남부현,나기상,박찬일 충남대학교 의과대학 지역사회의학연구소 1995 충남의대잡지 Vol.22 No.1

Widespread use of BCG has not controlled tuberculosis, and more effective vaccines are clearly needed. Although chemotherapy will remain the mainstay of antituberculosis treatment, the use of adjunctive immunotherapeutic modalitites is attractive, particularly in persons with drug-resistant tuberculosis. Administration of IL-2 or IFN-γto tuberculosis patients enhance bacillary elimination. Cell-mediated immunity is the critical protective immune response in tuberculosis. Mycobacterial antigens are recognized by T cells and that elicit production of protective cytokines are potentially important vaccine antigens. The 30 kDa antigen is secreted in large quantities by growing mycobacteria. That antigen elicits greater proliferation in lymphocytes from healthy tuberculin reactors than healthy tuberculin nonreactors. In this study, the T lymphocyte proliferative responses to 30 kDa antigen from Mycobactrium tuberculosis H37Rv were examined by using tonsilar and peripheral blood lymphocytes from PPD(+) and PPD(-) tonsilectomized persons. When cultured with 30 kD antigen, tonsilar mononuclear leukocytes and T cells of PPD(+) demonstrated more ^3H-thymidine incorporation than PPD(-) persons (stimulation index was 2.5 and 1.9, 0.8 and 1.0, repectively). Peripheral blood mononuclear cells (PBMC) and peripheral blood T lymphocytes were shown the similar responses to this antigen. The combination of IL-2 and 30 kDa antigen elicited a significant proliferative responsiveness in tonsilar mononuclear leukocytes and T cells of PPD(+) persons (SI was 20 and 14.1). PBMC and peripheral blood T cells of PPD(+) persons were also shown a significant responsiveness, but PPD(-) persons did not show. These results demonstrate that the 30 kDa antigen and IL-2 have a synergistic stimulatory property in mycobacteria sensitizing lymphocytes.

결핵균 30 kDa항원과 Interleukin-2 혹은 Anti-CD28의 병합이 건강인 말초혈액 림프구와 단핵구 증식에 미치는 영향

박정규,윤상호,조은경,김화중,백태현 충남대학교 의과대학 지역사회의학연구소 1994 충남의대잡지 Vol.21 No.2

Cell-mediated immunity is necessary for protection against Mycobacterium tuberculosis. T lymphocytes are thought to play a central role in cell-mediated immune response. And in vitro stimulation of leukocytes with mycobactria or their products induces synthesis and release of several cytokines, including IL-2. To study the T lymphocyte proliferative response to purified 30 kDa antigen from Mycobacterium tuberculosis H37Rv, IL-2 or their combination, peripheral blood lymphocytes and mononuclear cells isolated from healthy subjects were stimulated with the 30 kDa antigen, IL-2 or both. The proliferations of lymphocytes and molnonuclear cells to 30 kDa or 30/32 kDa antigen were significantly increased in PPD(+) group when compared to those in PPD(-) group. It is confirmed here that have shown synergy between 30 kDa antigen and IL-2(250U) in the induction of lymphocytes and mononuclear cells proliferation in PPD(+) group. The proliferative response reflects synergy between two separate activation stimuli. And macrophage-like cells existed in peripheral blood mononuclear cells were shown to help the synergy of the proliferative response to mycobacterial antigen and IL-2. The proliferative responses of lymphocytes and mononuclear cells to 30 kDa antigen were more increased than those to 30/32 kDa antigen. So it was suggested that 30 kDa antigen was the more immunogenic antigen than 30/32 kDa antigen.

Carbon tetrachloride를 투여한 rat의 hepatic lipid 축적에 미치는 vitamin E의 효과에 관한 연구

박은주,이경연,이미영,이외숙,장재정,정귀은,최진희 曉星女子大學校 藥學大學 學生會 1988 曉星藥誌 Vol.4 No.-

The present studies were undertaken to evaluate the effect of vitamin E, CCl_4 on the change of hepatic triglyceride, hepatic cholesterol, hepatic phospholipid in male rat. The result obtained from this study were summarised as follows: 1. Hepatic phospholipid of CCl_4 treated rat was increased in proportion to CCl_4 dosage but after concomitant injection(I.P) of vitamin E and CCl_4, hepatic phospholipid was significantly decreased in comparison to that of CCl_4 alone injection. 2. There was no effect on hepatic cholesterol concentration either CCl_4 alone injection(I.P) or concomitant injecton(I.P) of vitamin E and CCl_4. 3. Hepatic triglyceride of CCl_4-treated rat was significantly increased in comparison to that of normal rat but hepatic triglyceride of rat concomitant injection of vitamin E and CCl_4 was significantly decreased in comparison to that of CCl_4 alone injection.

구강점막 부착용 케토프로펜 고분자 필름의 제조 및 평가

박진석,이상은,강봉석,이경록,이은주,박정숙 충남대학교 약학대학 의약품개발연구소 2014 藥學論文集 Vol.29 No.-

Abstract – The objective of this study was to prepare ketoprofen-loaded buccal adhesive patch. The adhesive patch was formulated by casting method using aqueous soluble polymer povidone K17 (PVP 17PF) as film-forming agent and hydroxypropylmethylcellulose (HPMC) as adhesive agent. To compare the effect of HPMC type, different molecular weight of K4M and K15M HPMC was used. The physicochemical properties of patches such as appearance, thickness, in vitro release, and adhesiveness were investigated. The concentration of ketoprofen was determined spectrophotometrically at a wavelength of 233 nm. The appearance of prepared patches was semi-transparent, light-yellow or almost colorless, and odorless. Thickness of each patches (n=6) was 0.895 ± 0.033 mm for K4M patch and 0.727 ± 0.036 mm for K15M patch. In vitro release test, both K4M and K15M patches showed over 20% release within 30 min. At 120 min, K4M and K15M patches demonstrated 95% and 67.5% release of ketoprofen, respectively, and up to 240 min, both patches released drug completely. Maximum adhesive force of K4M and K15M patches was 6.571 ± 2.703 gf and 2.735 ± 1.151 gf, respectively. Moreover, it took 28.29 ± 0.38 sec and 28.30 ± 0.34 sec for K4M and K15M patch to peel off them after adhesion, showing no significant difference. In conclusion, thickness, in vitro release, and maximum adhesive force could be modulated by alteration of polymer types.

마우스 대장암 모델 구축 및 항암제 활성 평가를 위한 예비 연구

김예솔,강봉석,이상은,이은주,이경록,정상헌,박정숙 충남대학교 약학대학 의약품개발연구소 2014 藥學論文集 Vol.29 No.-

Abstract – Aberrant crypt foci (ACF) are early imorphological changes observed in rodents after administration of colon-specific carcinogen such as azoxymethane (AOM). ACF are considered to be putative preneoplastic lesions and are widely used as a surrogate biomarker to rapidly evaluate chemopreventive potential of compounds. The size of colorectal cancer was evaluated after administration of three anticancer drugs, 1 parent drug and 2 prodrugs. The body weights of mice were measured daily and considered as a surrogate for evaluation of general wellbeing. Colons were removed, cut along the longitudinal axis and flushed with phosphate-buffered saline. Each colon was cut into three equal lengths and fixed flat between filter papers. The fixed colon sections were stained with methylene blue. The number of ACF per colon, the number of aberrant crypts observed in each focus and the location of each focus were recorded. After single administration of AOM and multiple doses of anticancer drugs, no significant changes in the body weights of the mice was observed which was recorded for 52 days. However, an expected ACF was not observed in any treated groups. These findings suggest the induction of ACF in mice requires the promotion by dextran sulfate sodium as well as the initiation by AOM.

결핵균 30 kDa 단백항원 자극에 의하여 유도되는 IFN-Y 및 IL-4 mRNA 발현 양상과 세포독성능

백태현,조은경,박재하,박정규,김화중 충남대학교 생물공학연구소 1997 생물공학연구지 Vol.5 No.-

30 kDa protein antigen, a major secreted protein antigen of Mycobacterium tuberculosis, exhibits strong T cell stimulatory activity. In order to better understand the immunologic activities of 30 kDa antigen, lymphocyte proliferation by ^3H-thymidine incorporation, cytokine mRNA expression pattern using reverse transcription-polymerase chain reaction (RT-PCR), and cytotoxicity in response to in vitro stimulation of human peripheral blood mononuclear cells (PBMCs) with 30 kDa antigen were evaluated. Lymphoproliferative responses to 30 kDa and crude protein antigens in PBMCs of normal PPD-negative and positive donors were compared. As expected, PPD negative donors demonstrated no thymidine incorporation in response to 30 kDa and crude protein antigens, while PPD positive donors showed extensive proliferation to both antigens. Freshly isolated PBMCs were stimulated with 30 kDa antigen for 0, 6, 12, 24, 48 hr, 1 wk, and 2 wk and examined for the induction of IFN-γ and IL-4 mRNA using RT-PCR. The expression of IFN-γ mRNA was greatly augmented after 1 wk, and there was also early inductions at 6 and 24 hr, whereas IL-4 mRNA is consistently expressed through 0 to 48 hr and markedly decreased after 1 wk. In contrast, freshly isolated human tonsillar cells failed to express detectable level of IFN-γ mRNA but showed enhanced IL-4 mRNA expression after in vitro stimulation with 30 kDa antigen for 1 wk. Both natural killer (NK) and T cell cytotoxic activities induced by 30 kDa and crude antigens were also evaluated. PBMCs from PPD positive donors stimulated with 30 kDa antigen showed significantly higher NK and T cell cytotoxic activities than those in PPD negative donors. Crude antigen also induced similar level of cytotoxicity with that of 30 kDa antigen. These results suggest that 30 kDa antigen of M. tuberculosis may selectively activate Th1 cells and be a strong inducer of IFN-γ expression. In conclusion, 30 kDa antigen can be used as a standard T cell immunogen.

MTT비색법에 의한 결핵균 30-kDa항원이 임파구 증식에 미치는 영향

백태현,김민경,박정규,김화중,조은경,최대경 충남대학교 의과대학 지역사회의학연구소 1991 충남의대잡지 Vol.18 No.1

The cell mediated immune response appears to result from the specific recognition of an antigen. by T lymphocytes. It has been well recognized that T cell play an important roles in the induction of tuberculin hypersensitivity and immunity to tuberculosis. In order to evaluate effect on T lymphocyte proliferative response to purified 30-kDa antigen from Mycobacterium tuberculosis, stimulation effects of peripheral blood lymphocytes with 30-kDa antigen, crude antigen and PHA were measured by MTT colorimetric assay. Three groups of healthy subjects, representing PPD(+), PPD() and PPD(-) persons, were investigated. The proliferative response to 30-kDa antigen were elicited the plateau at concentration of 20 to 0.1㎍/ml, crude antigen showed rapid reduction as decreasing the concentration of antigen, for the PPD(+) person. Both antigens, at concentration of <1㎍/ml, failed to stimulate lymphocytes of the PPD(-) person. Therefore the concentration of 30-kDa antigen to induce optimal stimulation was 1㎍/ml. The lymphocyte proliferation to 30-kDa antigen and crude antigen were significantly increased in PPD(+) group when compared to those in PPD(-) group, but PHA response was no significant difference. These results suggest that 30-kDa antigen could stimulate lymphocyte from PPD(+) population and MTT colorimetric assay could be applied to assess proliferative response of lymphocyte.