Earticle

Postnatal rat offspring born from prenatal alcohol-intoxiciated pups during pregnancy leads to apoptosis or inhibitation of neurigenesis, which eventually causes neurological impairments. In the present study, the effects of endurance exercise on the level of apoptosis and cell proliferation in the hippocampal dentate gyrus following fetal alcohol syndrome in Sprague-Dawley rats were investigated. Adult female Sprague– Dawley rats weighing 220±10 g were used in the experiment. Total 60 female rats were mated in pairs with adult 30 male Sprague–Dawley rats weighing 300±10g for 48h. After mating, the female rats were housed individually in a plastic home cage. After confirmation of pregnancy, the pups borned from female rats were randomly divided into four groups: the control group, fetal alcohol-treated group, fetal alcohol-treated and moderate-intensity exercise group, and fetal alcohol-treated and swimming exercise group(n=20, respectively). The exercise groups were made to run on a treadmill running and swimming for 60 min per day for 14 consecutive days. Apoptotic cell death in the hippocampal dentrate gyrus was measured by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) and caspase-3 staining, and cell neurogenesis was detected by 5-bromo-2'-deoxyuridine(BrdU) immunohistochemistry. It was shown that apoptosis and cell proliferation in the hippocampal dentate gyrus were significantly increased following prenatal alcohol injection in Sprague–Dawley rats and that treadmill exercise and swimming suppressed the alcohol-induced increase of apoptosis and cell neurogenesis in the dentate gyrus. The present results suggest that treadmill exercise and swimming may protect neurons from apoptotic death and aids in recovery from the central nervous system sequelae following fetal alcohol syndrome.

본 연구의 목적은 정신장애인 기능평가척도의 단축형(FAS-S)을 개발하고 타당화하는 것이다. 연구 1은 척도의 요인구조를 파악하기 위한 연구이다. 연구 1에는 총 640명의 정신장애인(남자 410명, 여자 230명)이 참여하였다. 탐색적 요인분석 결과 8개의 요인(요인당 6문항)이 추출되었다. 이들 요인들은 위생/건강관리, 병식과 도움요청, 생활문제해결기술, 대화기술, 가사관리, 과업수행, 대인적 문제행동, 대인관계이다. 연구 2는 타당화 연구이다. 연구 2에는 정신장애인 506명(남자 306명, 여자 200명)이 참여하였다. 확인적 요인분석 결과, 6요인, 요인 당 4문항으로 구성된 척도의 전반적인 부합지수가 가장 높은 것으로 나타났다. 2차 요인 분석 결과, 6개의 1차 요인들이 1개의 2차 요인으로 수렴되는 모형보다 2개의 2차 요인으로 수렴되는 모형의 부합도가 더 좋은 것으로 나타났다. 따라서 본 연구에서 개발된 척도는 2개의 하위 척도로 사용할 수 있음이 시사된다. 본 연구에서 개발된 척도와 다른 척도들간의 상관분석을 통하여 기능평가척도의 동시타당도를 검토한 결과, 2개의 기능평가 하위척도 중 일상생활 기능척도는 사회적 기능척도보다 증상과 더 큰 부적 상관을 보였다. 사회적 기능척도는 일상생활 기능척도보다 재기태도, 희망, 삶의 질, 사회적 지지와 더 높은 상관을 보였다. 연구의 시사점과 추후 연구방향이 논의되었다.

This study aimed to develop and validate a short form Functional Assessment Scale for persons with psychiatric disabilities (FAS-S) and to explore the factor structure of the FAS-S. Participants in Study 1 were 640 persons with psychiatric disabilities (410 males and 230 females). Participants' mean age was 38.1 (SD = 9.47). Exploratory factor analysis produced eight factors: (1) hygiene/health care, (2) insight/help seeking, (3) problem solving, (4) conversation skills, (5) household management, (6) task performance, (7) problematic behavior, and (8) interpersonal relationships. Study II, to validate the FAS-S, enrolled 506 participants (306 males and 200 females), who had suffered from psychiatric disorders for a mean period of 11.92 years (SD = 8.13). Confirmatory factor analysis using 24 items resulted in favorable goodness of fit indices (Chi-square = 416.5, df = 236, p < .001, GFI=.930, AGFI=.910, NFI = .924, NNFI = .958, CFI = .964, RMSEA = .041). Secondary factor analysis results showed two secondary factors (factor 1: “activities of daily life [DAL]”; factor 2: social skills) had favorable goodness of fit indices (X2 = 506.0, df = 244,p < .001, GFI = .916, AGFI = .897, NFI =.910, NNFI=.945, CFI = .951, RMSEA = .048). In addition, we calculated concurrent validities between the FAS-S sub-scales and certain other scales. The results showed that the DAL subscale of the FAS-S had a significant negative correlation to the PANSS, while the social skills subscale score of the FAS-S had significantly higher positive correlations with recovery attitude, hope, quality of life, and social support than did the DAL subscale score. We discuss the implications of these results and applications of the FAS-S.

Fas gene known to associate with intramuscular fat content in Korean cattle was selected for DNA marker development. Fas (APO-1, CD95), a member of the tumor necrosis factor(TNF) receptor superfamily, is a cell membrane protein that mediates apoptosis(programmed cell death). We discovered single nucleotide polymorphisms(SNPs) within Fas gene in order to develop novel DNA markers at genomic level. Of this gene to search for SNP, sequences of whole exon and 1kb range of both front and back of the gene using 24 cattle were determined by direct-sequencing methods. As a result, 16 SNPs in exon, 37 SNPs in intron and 2 SNPs in promoter region, a total of 55 SNPs were discovered. In these SNPs, thirty-one common polymorphic sites were selected considering their allele frequencies, haplotype-tagging status and Linkage Disequilibrium(LD) for genotyping in larger-scale subjects. Selected SNPs were confirmed genotype through SNaPshot method(n=274) and were examined for possible genetic association of Fas polymorphisms with carcass weight(CWT), eye muscle area(EMA), and backfat thickness(BF). So, the SNP have been identified significant g.-12T>G, g.1112T>G and g.32548T>C. These results suggest that polymorphism of Fas gene was associated with meat quality traits in Hanwoo.

Several human leukocyte subsets including natural killer (NK) cells, cytotoxic T lymphocytes (CTL), and polymorphonuclear neutrophils (PMN) participate in cellular immune responses directed against vascularized pig-to-human xenografts. As these leukocytes express the death receptor Fas either constitutively (PMN) or upon activation (NK, CTL), we explored in vitro whether the transgenic expression of membrane-bound human Fas ligand (mFasL) on porcine fetal fibroblasts is a valuable strategy to protect porcine xenografts. cDNA of mFasL carrying the deletion at the cleavage site with metalloproteinase and lacking the death domain in its cytoplasmic tail was subcloned into pCAGGS expression vector driven by the chicken β-actin promoter containing blastidin- resistance cassette. The mFasL expression vector was transfected into mini-pig fetal fibroblasts by lipofection method. Blastidin-resistant cells were screened by PCR and FISH. The expression of mFasL was confirmed by Western blot and FACS with the mouse anti-human FasL antibody. Interaction of two transgenic clonal cell lines with human leukocytes was analyzed using functional assay for cytotoxicity. mFasL expressed on porcine fetal fibroblasts protected porcine fetal fibroblasts against killing mediated by human NK cells. The rate of NK cell mediated cytotoxicity was significantly reduced in transgenic clonal cells (54±10.80%) compared to normal minipig fetal fibroblasts. This result indicated that grafts of transgenic pigs expressing mFasL could control the cellular immune response to xenografts, and create a window of opportunity to facilitate xenograft survival.

The main goal of this research is to achieve development of recombinant E. coli for fatty acid synthesis(FAS). Genes for pyruvate dehydrogenase (aceE) and acetyl-CoA carboxylase (accA), which are the enzymes that catalyze the first step in the synthesis of fatty acids in Escherichia coli MG1655, were cloned and characterized. That is, the E. coli strains were midterm recombinant strains to produce fatty acid. The enzyme of aceE gene converts pyruvate to acetyl-CoA and that of accA gene catalyzes the addition of CO2 to acetyl-CoA to generate malonyl-CoA. The genes were identified as homologous gene of E. coli through a metabolic pathway. The recombinant E. coli MG1655 containing the aceE or accA gene-inserted expression vector(pTrc99A) were constructed. As a result, the intermediate products were analyzed from in vivo metabolites and in vitro metabolites of recombinant E. coli. The two strains produced more malonic acid than its wild type E. coli. However, there was insignificant the difference between E. coli harboring a aceE gene-inserted pTrc99A vector and E. coli harboring a accA gene-inserted pTrc99A vector.

The microbial biosynthesis of fatty acid, which can be used as precursors for the production of fuels of chemicals from renewable carbon sources, has attracted significant attention in recent years and related fields have been actively studied. To produce fatty acid from bacteria, the functions of enzymes involved in fatty acid synthesis and proteins encoded by related genes on the fatty acid synthesis pathway have been also studied in the FAS system of bacteria. Therefore, the goal of this study was to increase the production of long-chain fatty acids by developing recombinant E. coli strains that were improved by an elongation cycle during fatty acid synthesis (FAS). The fabF (β-oxoacyl-[acyl-carrier-protein] synthase II), fabG (β- oxoacyl-[acylcarrier- protein] reductase), fabZ (β-hydroxyacyl-[acyl-carrier-protein] dehydratase) and fabI (enoyl-[acyl-carrier-protein] reductase) genes, all homologous of E. coli, were induced to improve the enzymatic activities for the purpose of over- expressing components of the elongation cycle in the FAS pathway through metabolic engineering. The developed E. coli strains contained various gene combinations with the pSTV28 expression vector. In vivo productivity of total fatty acids was analyzed to confirm the changes and effects of inserted genes in E. coli at different incubation temperatures. As a result, total fatty acid contents including hexadecanoic acid and octadecanoic acid such as saturated fatty acid and unsaturated fatty acid were produced higher in developed strains than those from the wild-type E. coli. Especially total fatty acid was produced 2.4 fold higher than E. coli JESH1007, one of the developed reombinants through this study, at 20℃.

The goal of this study was to increase fatty acid production by developing recombinant E. coli strains which fatty acid synthesis (FAS) were improved. The acetyl-CoA carboxylase (ACC) enzyme from accA, accB and accC genes catalyzes the addition of CO2 to acetyl-CoA to generate malonyl-CoA. The enzyme encoded by the fabD gene converts malonyl-CoA to malonyl-[acp], and the EC3.1.2.14 gene converts fatty acyl-ACP chains to long chain fatty acids. Therefore, over-expression of acc family genes and EC3.1.2.14 gene was expected to increase the productivity of fatty acids. All recombinant E. coli strains containing various gene combinations to enhance the enzymatic activities were developed using the pTrc99A vector. The in vitro metabolites, amount of total lipid and fatty acids produced were analyzed to observe the changes in metabolism by introducing five distinct genes. This result indicated that from recombinant E, coli which had EC3.1.2.14 gene, the amount of total lipid and fatty acids were produced much more than the wild type E. coli.

The goal of this research is to develop recombinant E. coli for improved fatty acid synthesis (FAS). Genes for acetyl-CoA carboxylase (accA, accB, accC), malonyl-CoA-[acyl-carrier-protein] transacylase (fabD), and acyl-acyl carrier protein thioesterase (E3.1.2.14), which are the enzymes that catalyze the key steps in the synthesis of fatty acids, were cloned and over-expressed in E. coli MG1655. The enzymes of acc family genes catalyze the addition of CO2 to acetyl-CoA to generate malonyl-CoA, enzyme of fabD gene converts malonyl-CoA to malonyl-[acp], and gene for E3.1.2.14 converts fatty acyl-ACP chain to long chain fatty acids. All the genes were identified as homologous gene of E. coli and used to improve enzyme activities for overflowing of FAS pathway through a metabolic engineering. Eleven different E. coli MG1655 strains containing various combination of genes were developed by using pTrc99A expression vector. To observe changes in metabolism, in vivo and in vitro metabolites and fatty acids were analyzed from recombinants. The recombinant strains produced more malonic acid and fatty acids than its wild type did.

The goal of this research is to achieve development of recombinant E. coli for fatty acid synthesis (FAS). The enzyme of aceE gene converts pyruvate to acetyl-CoA and that of accA gene catalyzes the addition of CO2 to acetyl-CoA to generate malonyl-CoA. The genes were identified as homologous gene of E. coli through a metabolic pathway. The genes for pyruvate dehydrogenase (aceE) and acetyl-CoA carboxylase (accA), which are the enzymes that catalyze the first step in the synthesis of fatty acids in Escherichia coli MG1655, were cloned. That is, the E. coli strains were midterm recombinant strains to produce fatty acid. The recombinant E. coli MG1655 containing the aceE or accA, and the two genes inserted expression vector (pTrc99A) were developed. To confirm effects of each gene, the intermediate products were analyzed from in vivo metabolites and in vitro metabolites of recombinant E. coli. As a result, the three strains produced more malonic acid than its wild type E. coli. However, there was insignificant the difference between E. coli harboring a aceE gene-inserted pTrc99A vector and E. coli harboring a accA gene-inserted pTrc99A vector.

=목적: 본 연구는 정상임신, 전자간증 및 HELLP 증후군 임산부의 태반에서 세포사멸 면역 조절인자인 Fas 및 Fas 배위자의 태반 내 발현을 비교하고, 전자간증 및 HELLP 증후군 임산부의 혈청 내 Fas 및 Fas 배위자의 변화를 관찰하기 위하여 본 연구를 시행하였다.연구 방법: 정상 단태 임신부 31명, 전자간증 임신부 27명, HELLP로 변환된 전자간증 5명을 대상으로 혈청 샘플을 얻어 Enzyme linked immunoassay에 의한 방법으로 결과를 얻었으며, 태반에서 세포사멸을 관찰하기 위하여 광학 현미경 및 Fas/Fas ligand에 대한 면역조직화학 염색을 시행하였다. Mann-Whitney test, χ2 test, Pearson correlation coefficients와 multiple regression test를 통계에 이용하였다.결과: 광학현미경적 관찰에서 태반 내 세포사멸을 정상임신, 전자간증 모두에서 관찰할 수 있었다. 또한 면역조직화학염색을 이용하여 세포사멸 관련 단백질 발현을 영양막 세포에서 조사한 결과 Fas 단백의 발현은 전자간증 군에서 정상임신에 비해 증가하는 양상을 보였고, Fas 배위자 단백의 발현은 전자간증 군에서 정상임신에 비해 발현이 감소하는 양상을 보였다. 그리고 Fas 배위자 단백의 발현은 Fas 단백에 비해 발현의 강도가 미약하였다. Fas의 평균값은 정상 임산부의 경우 5.83±0.37 U/mL, 전자간증 임산부의 경우 10.84±0.93 U/mL, HELLP 증후군 임산부의 경우는 10.79±0.69 U/mL이었으며, Fas ligand의 평균값은 정상 임산부의 경우 0.59±0.03 U/mL, 전자간증 임산부의 경우 0.51±0.21 U/mL, HELLP 증후군 임산부의 경우는 0.60±0.01 U/mL이었다. 따라서 전자간증 임신과 HELLP로 변환된 전자간증에서 soluble Fas값이 정상 단태 임신보다 통계학적으로 유의하게 높았으나 (P<0.01), Fas ligand의 경우 각 군간의 차이는 없었다.

=목적: 자연 폐경된 여성 229명을 대상으로 Fas와 Fas-ligand 유전자의 다형성에 따른 골밀도의 연관성과 1년간 호르몬요법 후 골밀도를 측정하여 골 반응도와 유전자 다형성간 연관성을 분석하였다. 연구 대상 및 방법: Fas와 Fas-ligand 유전자의 다형성은 RFLP방법을 이용하여 분석하였으며, 호르몬요법 전과 1년 후 요추, 대퇴골경부, Ward 삼각주, 대퇴돌기의 골밀도를 측정하여 Fas와 Fas-ligand 유전자의 다형성에 따른 골밀도의 경향과 1년간 호르몬요법 후 골 반응도와 유전자 다형성간 연관성을 분석하였다. 결과: Fas 유전자의 A670G와 G1377A 다형성은 요추에서는 AA 유전자형, 다른 부위에서는 GG 유전자형, 그리고 Fas-ligand 유전자의 C843T 다형성은 요추 부위에서는 T/T 유전자형, 대퇴돌기 부위에서는 나머지 유전자형 (C/C, C/T)이 낮은 골밀도와 깊은 연관성이 있었다. Fas 유전자의 A670G의 A/A 유전자형, G1377A의 G/G 유전자형, 그리고 Fas-ligand 유전자의 C843T의 T/T 유전자형은 호르몬요법 후 유의한 골밀도 변화가 관찰되었다.

사람 난소내 난포의 수적 감소와 배란 후 황체의 퇴축 과정은 apoptosis에 의해서 이루어진다. 그러나 이러한 생리적 과정에 어떤 인자들이 영향을 주는지는 아직 밝혀져 있지 않다. 따라서 저자들은 난소내 apoptosis와 이에 영향을 끼치는 인자들의 관계를 규명하기 위하여 Fas, Fas-ligand, Bcl-2 및 Bad 항체를 이용한 면역조직화학적 방법으로 사람 난소의 원시 난포, 일차 난포, 우성 난포에서의 발현을 관찰하였다.Fas의 발현은 원시 난포는 난자에서만, 일차 난포는 난자와 과립막 세포에서 그리고 우성 난포의 경우 난자, 과립막 세포와 난포막 세포 모두에서 관찰되었으며 황체의 경우는 퇴축되면서 발현이 감소하였다. Fas- ligand의 발현은 퇴화된 난포의 난자와 과립막 세포 그리고 황체에서 관찰되었고 황체 퇴축 과정이 진행되면서 염색이 감소하였다. Bcl-2의 발현은 일차 난포와 우성 난포의 난자와 과립막 세포 그리고 황체에서 관찰할 수 있었으며 난포가 성장할수록 발현이 증가하였으나 퇴화된 난포에서는 관찰할 수 없었다.Bad의 발현은 성장하는 난포 세포들에서 일차 난포의 과립막 세포에서만 약하게 발현이 관찰되었으나 퇴화된 난포에서는 일차 난포와 우성 난포에서 모두 발현이 관찰되었다. 그리고 폐경기 난소에서는 모든 항체에 대한 발현을 관찰할 수 없었다.

Expression of Fas, Fas-ligand, Bcl-2 and Bad with Maturationof Human Ovarian Follicle