• BMB Reports
• /
• 제44권6호
• /
• pp.405-409
• /
• 2011

The present study demonstrated that valproic acid (VPA) transcriptionally regulates human GM3 synthase (hST3Gal V), which catalyzes ganglioside GM3 biosynthesis in ARPE-19 human retinal pigment epithelial cells. For this, we characterized the promoter region of the hST3Gal V gene. Functional analysis of the 5'-flanking region of the hST3Gal V gene revealed that the -177 to -83 region functions as the VPA-inducible promoter and that the CREB/ATF binding site at -143 is crucial for VPA-induced expression of hST3Gal V in ARPE-19 cells. In addition, the transcriptional activity of hST3Gal V induced by VPA in ARPE-19 cells was inhibited by SP600125, a c-Jun N-terminal kinase (JNK) inhibitor. In summary, our results identified the core promoter region in the hST3Gal V promoter and for the first time demonstrated that ATF2 binding to the CREB/ATF binding site at -143 is essential for transcriptional activation of hST3Gal V in VPA-induced ARPE-19 cells.

• 생명과학회지
• /
• 제14권6호
• /
• pp.1009-1017
• /
• 2004

인간유래 시알산전이효소 유전자들의 특이적 발현과 그들의 mRNA isoform의 생성에 대한 조절기구를 이해하기 위하여 5종류의 human 시알산전이효소 유전자(hST3Cal II, hST8Sia II, hST8Sia III, hSTS8Sia IV, hST8Sia V)들의 게놈구조를 분석하였다. hST3Gal II 유전자는 17 kb이상의 게놈상에 46 bp에서 1017 bp의 길이를 가진 exon이 6개로 이루어져 있고, hST8Sia III유전자는 10 kb이상의 게놈상에 125bp에서 2023bp의 길이를 가진 exon이 4개로 이루어져 있어 다른 human 시알산전이효소 유전자들보다 짧고 단순한 구조를 가지고 있었다. 반면에 다른 3종류의 유전자(hST8Sia II, hST8Sia IV, hST8Sia V)들은 70 kb이상의 게놈상에 5개이상의 exon으로 이루어져 있으며, 5종류 모두 exon-intron boundary는 GT-AG rule을 나타내고 있었다. 특히 모든 시알산전이효소에 고도로 보존되어 있는 sialylmotif L은 hST8Sia III유전자에서는 하나의 exon에 존재하는 반면에, 다른 시알산전이효소 유전자에서는 분리된 exon에 존재하여 exon의 구조적 다양성을 나타내고 있다. 또한, 본 연구에서는 5'-RACE와 cap site hunting법에 의해 hST3Gal II 유전자의 전사개시점을 결정하였다.

• Reproductive and Developmental Biology
• /
• 제35권4호
• /
• pp.511-518
• /
• 2011

To avoid hyperacute rejection of xenografts, ${\alpha}1,3$-galactosyltransferase knock-out (GalT KO) pigs have been produced. In this study, we examined whether Sia-containing glycoconjugates are important as an immunogenic non-Gal epitope in the pig liver with disruption of ${\alpha}1,3$-galactosyltransferase gene. The target cells were then used as donor cells for somatic cell nuclear transfer (scNT). A total of 1,800 scNT embryos were transferred to 10 recipients. One recipient developed to term and naturally delivered two piglets. Real-time RT-PCR and glycosyltransferase activity showed that ${\alpha}2,3$-sialyltransferase (${\alpha}2,3ST$) and ${\alpha}2,6$-sialyltransferase (${\alpha}2,6ST$) in the heterozygote GalT KO liver have higher expression levels and activities compared to controls, respectively. According to lectin blotting, sialic acidcontaining glycoconjugate epitopes were also increased due to the decreasing of ${\alpha}$-Gal in heterozygote GalT KO liver, whereas GalNAc-containing glycoconjugate epitopes were decreased in heterozygote GalT KO liver compare to the control. Furthermore, the heterozygote GalT KO liver showed a higher Neu5Gc content than control. Taken together, these finding suggested that the deficiency of GalT gene in pigs resulted in increased production of Neu5Gc-bounded epitopes (H-D antigen) due to increase of ${\alpha}2,6$-sialyltransferase. Thus, this finding suggested that the deletion of CMAH gene to the GalT KO background is expected to further prolong xenograft survival.

• Journal of Acupuncture Research
• /
• 제17권2호
• /
• pp.95-117
• /
• 2000

The purpose of this morphological studies was to investigate the relation between the meridian, acupoints and viscera using neuroanatomical tracers. The common locations of the spinal ganglia, sympathetic chain ganglia, spinal cord and brain projecting to the large intestine meridian were observed following injection of transganglionic tracer, WGA-HRP and transsynaptic neurotropic virus, pseudorabies virus(PRV), Bartha strain(Ba) and PRV-Ba-Gal (Galactosidase)) into the the large intestine(cecum, colon and rectum), ST37 and LI4. After survival times of 96 hours following injection into the thirty rats with WGA-HRP, PRV-Ba and PRV-Ba-Gal. They were perfused, and their spinal ganglia, sympathetic chain ganglia, spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by HRP and X-gal histochemical and PRV immunohistochemical staining method, and observed with a light microscope. The results were as follows : 1. WGA-HRP labeled neurons innervating the large intestine were observed bilaterally within the T13-L4 sympathetic chain ganglia, and T9-11 spinal ganglia. WGA-HRP labeled neurons innervating ST37 were observed within the L3-5 sympathetic chain ganglia, and L2-4 spinal ganglia. WGA-HRP labeled neurons innervating LI4 were observed in the middle cervical ganglion and stellate ganglion, and C5-8 spinal ganglia. 2. In spinal cord, PRV-Ba labeled neurons projecting to the large intestine, ST37 and LI4 were found in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina N, V, VII(intermediolateral nucleus), Ⅸ, X and dorsal nucleus. 3. In medulla oblongata, PRV-Ba and PRV-Ba-Gal labeled neurons projecting to the large intestine, ST37 and LI4 were commonly found in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus and gigantocellular nucleus. 4. In pons, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in locus coeruleus, Kolliker-Fuse nucieus and A5 cell group. 5. In midbrain, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in central gray matter. 6. In diencephalon, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in paraventricular hypothalamic nucleus. These results suggest that PRV-Ba and PRV-Ba-Gal labeled common areas projecting to the large intestine may be correlated to that of the large intestine meridian, ST37 and LI4. Especially, These morphological results provide that interrelationship of meridian-acupoints -viscera may be related to the central autonomic pathways.

• 한국수정란이식학회지
• /
• 제30권4호
• /
• pp.319-325
• /
• 2015

Gangliosides exist in glycosphingolipid-enriched domains on the cell membrane and regulate various functions such as adhesion, differentiation, and receptor signaling. Ganglioside GM3 by ST3GAL5 enzyme provides an essential function in the biosynthesis of more complex ganglio-series gangliosides. However, the role of gangliosides GM3 in porcine oocytes during in vitro maturation and early embryo development stage has not yet understood clear. Therefore, we examined ganglioside GM3 expression patterns under apoptosis stress during maturation and preimplantation development of porcine oocytes and embryos. First, porcine oocytes cultured in the NCSU-23 medium for 44 h after $H_2O_2$ treated groups (0.01, 0.1, 1 mM). After completion of meiotic maturation, the proportion MII (44 h) was significantly different among control and the H2O2 treated groups ($76.8{\pm}0.3$ vs $69.1{\pm}0.4$; 0.01 mM, $55.7{\pm}1.0$; 0.1 mM, $38.2{\pm}1.6%$; 1 mM, P<0.05). The expressions of ST3GAL5 in $H_2O_2$ treated groups were gradually decreased compared with control group. Next, changes of ST3GAL5 expression patterns were detected by using immunofluorescene (IF) staining during preimplantation development until blastocyst. As a result, we confirmed that the expressions of ST3GAL5 in cleaving embryos were gradually decreased (P<0.05) according to the early embryo development progress. Based on these results, we suggest that the ganglioside GM3 was used to the marker as pro-apoptotic factor in porcine oocyte of maturation and early embryo production in vitro, respectively. Furthermore, our findings will be helpful for better understanding the basic mechanism of gangliosides GM3 regulating in oocyte maturation and early embryonic development of porcine in vitro.

• Journal of Acupuncture Research
• /
• 제20권6호
• /
• pp.89-102
• /
• 2003

Objective: The expression of CRF(corticotropin releasing factor), CRF-R(receptor) and CRF-BP(binding protein) in CNS neurons projecting to the stomach and ST36 using the pseudorabies virus in the rat was investigated. Methods: After survival times of 5 days following injection of PRV-Ba-Gal, The thirty rats were perfused, and their brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV-Ba-Gal histochemical staining method and(or) CRF, CRF-R and CRF-BP immunohistochemical method. The common expressed areas of the brain projecting to the stomach and zusanli(ST36) following injection of PRV-Ba-Gal were observed with light microscope. Results: 1) The dense accumulation of CRF-immunoreactive terminals is seen in the area postrema, n. tractus solitarius, external zone of the median eminence, with some immunoreactive CRF also present in the internal zone. 2) Aggregates of CRF-R immunoreactive perikarya are found in area postrema, n. tractus solitarius, lateral reticular n., gigantocellular reticular n., locus coeruleues, paraventricular n. of hypothalamus, median eminence, preoptic n., arcuate n. and hind limb area of cerebral cortex. 3) Aggregates of CRF-BP immunoreactive perikarya are found in area postrema, n. tractus solitarius, lateral reticular nucleus, gigantocellular reticular n., locus coeruleues, paraventricular n. of hypothalamus, median eminence and arcuate n.. Conclusions : These results suggest that PRV-Ba-Gal labeled areas projecting to stomach and ST36 may be related to the central autonomic pathways. A part of CNS neurons projecting to the stomach and ST36 were related to expression of CRF, CRF-R and CRF-BP related to the stress in central autonomic center.

• 한국생명과학회:학술대회논문집
• /
• 한국생명과학회 2001년도 10주년 기념 국제학술 심포지움 및 추계학술대회
• /
• pp.47-47
• /
• 2001

• 약학회지
• /
• 제51권2호
• /
• pp.96-102
• /
• 2007

Measurement of globotriaosylceramide (Gb3, ceramide trihexoside) in urine has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. A simple, rapid, and highly sensitive analytical method for Gb3 in urine was developed without labor-extensive pre-treatment by electrospray ionization MS/MS (ESI-MS/MS). Only simple 5-fold dilution of urine is necessary for the extraction and isolation of Gb3 in urine. Gb3 in diluted urine was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Eight isoforms of Gb3 were completely resolved from urine matrix. C24:0 Gb3 occupied 50% of total Gb3 as a major component in urine. Linear relationship for Gb3 isoforms was found in the range of 0.005${\sim}$5.0 ${\mu}$g/ml. The limit of detection (S/N=5) was 0.005 ${\mu}$g/ml and limit of quantification was 0.05 ${\mu}$g/ml for C24:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9598 to 0.9975. This method could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

• 한국조리학회지
• /
• 제15권2호
• /
• pp.108-120
• /
• 2009

본 연구는 청소년들의 패스트푸드 메뉴의 선택 속성 및 만족도를 알아보기 위한 연구로, 경남 창원시의 고등학교에 재학 중인 학생들을 대상으로 실시하였다. 총 400부의 설문지 중 306부의 유효한 자료만을 분석에 사용하였으며, 데이터 코딩 과정을 거친 후 SPSS 12.0을 활용하였다. IPA 분석 결과 1사분면에 8개, 2사분면에 6개, 3사분면에 5개, 4사분면에 8개의 변수가 포함되었으며, 요인 분석 결과 총 5개의 요인(분위기, 음식, 종사원, 편안함, 보상)으로 나타났다. 이들 요인 중 '음식' 요인만이 패스트푸드 만족도에 유의적인 영향을 미치는 것으로 나타났다.

• 대한미생물학회지
• /
• 제3권1호
• /
• pp.35-41
• /
• 1968

한국(韓國)에는 매년(每年) 상당(相當)한 수(數)의 장(腸)지브스유행(流行)이 발생(發生) 되고있으나 대부분(大部分)의 경우(境遇) 기기록(其記錄)이 없거나 혹(惑)은 지극(至極)히 미비(未備)한 사실(事實)을 부인(否認)할수가 없다. 1959년(年)에 김상태(金相泰) 고상균(高相均)은 1957년(年)에 발생(發生)된 경북(慶北)의 16례(例), 서울 맹아학교(盲啞學校)의 70례(例), 부산(釜山)의 60례(例), 묵호(墨湖)의 245례등(例等)과 1958년(年)에 발생(發生)된 전북연도(全北煙島)의 60례(例)를 요약(要約)한바 있으나 거창(居昌)(1954년(年)) 함양(咸陽)(1965년(年))및 단양(丹陽)(1966년(年)) 등(等)의 대유행(大流行)에 관(關)하여서는 별(別)로 기록(記錄)되어 있는바가 없다. 이러한 점(點)에 비추어 저자등(著者等)은 1967년(年)에 경남삼천포시(慶南三千浦市)에 발생(發生)된 장(腸)지브스 유행(流行)에 관(關)하여 조사(調査)한바 있으므로 이에 기록(記錄)하는 바이다. 1. 인구(人口) 54,064의 삼천로시(三千浦市)에 638명(名)의 환자(患者)가 발생(發生)하여 1,180/100,000이라는 고율(高率)의 이환율(罹患率)을 나타냈고 사망(死亡)은 1명(名)이었다. 2. 이 유행(流行)은 1967년(年) 9월초(月初)에 시작(始作)되어 동년(同年) 11월말(月末)에 종식(終熄)되었으나 최고환자발생시기(最高患者發生時期)는 9월중(月中) 제3주(第3週)였으며 계연유행곡선(繼燃流行曲線)은 10월중순(月中旬)까지 계속(繼續)되었다. 3. 감염경로(感染經路)는 수인성(水因性)이었으며 감염원(感梁源)은 진구(陳舊)된 정호(井戶)(갈때샘))이었다. 4. 원인균(原因菌)은 Salmonella typhi로 동정(同定)되었다. 5. 모든 환자(患者)와 보균자(保菌者)는 삼천포시(三千補市) 의사회(醫師會)의 감독하(監督下)에 재가치료(在家治療)되었다. 6. 조사기중(調査期中)에 1년전(年前)에 장(腸)지브스에 이환(罹患)되었든 3명중(名中)에서 1명(名)이 보균자(保菌者)로 나타났으며 Shigella flexneri에 의(依)한 이질환자(痢疾患者)도 1명(名) 검출(檢出)되었다. 7. 1967년(年) 11월(月) 삼천포시유행(三千補市流行)이 거의 끝날무렵 인접고성군(隣接固城郡)에도 장(腸)지브스발생(發生)이 보고(報告)되었으며 역시(亦是) Salmonella typei가 검출(檢出)되었다. 8, 1968년(年) 춘계(春季)에 실시(實施)된 전년도환자(前年度患者)에 대(對)한 보균자검색(保菌者檢索)에서 1건(件)의 보균자(保菌者)도 검출(檢出)되지 않았으며 1968년(年) 10월(月)까지의 보고(報告)에서는 1건(件)의 신환(新患)도 발생(發生)되지 않았다.