• Title/Summary/Keyword: agglutinin

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Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

Clinical Findings of Mycoplasma Pneumonia in Children, from 1998 to 2003 (최근 6년간(1998-2003) 소아 Mycoplasma 폐렴의 임상 양상)

  • Kim, Ji-Hyun;Chae, Soo Ahn;Lee, Dong-Keun
    • Clinical and Experimental Pediatrics
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    • v.48 no.9
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    • pp.969-975
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    • 2005
  • Purpose : We performed a study of clinical findings of Mycoplasma Pneumonia in children, to know differences between recent clinical manifestations of Mycoplasma pneumonia and previous studies. Methods : The subjects of this study were 393 children who were diagnosed as Mycoplasma pneumonia with high titers of Mycoplasma antibody(${\geq}1$ : 160) or fourfold rises of Mycoplasma antibody at Chung Ang University Hospital from January 1998 to December 2003. We practiced a retrospective study on the clinical manifestations of Mycoplasma pneumonia based on their medical records. Results : Male to female ratio was 1.06 to 1 and mean age was $4.32{\pm}2.94years$. The highest incidence was in the age of 2 to 3 years(18.6 percent). Most frequent months were October, and November in 2000, April in 2002, and October and, December in 2003. Twenty six point seven percent showed allergic diseases. Second degree relatives of 10.7 percent patients had allergic diseases. Forty three point three percent were admitted before this admission for pneumonia. Allergic tests were positive in 65.7 percent. Cough, and sputum were the most common symptoms and abdominal pain, and vomiting were the most frequent extrapulmonary symptoms. Atelectasis and pleural effusion were seen in 2.5 percent and 1.8 percent. Infiltrations were more common on the right side. Titers of each simultaneous test for cold agglutinin and mycoplasma antibody were not in proportion to each other (P=0.163). Conclusion : The onset age of mycoplasma pneumonia was found to be lower than it used to be. More patients had a past medical history or a family history of allergic disease, and more wheezing was heard and associated with recurrent pneumonia.

Clinical Characteristics of Mycoplasma pneumoniae Pneumonia in Children during Recent 10 Years (최근 10년간 Mycoplasma pneumoniae 폐염의 임상양상에 대한 고찰)

  • Lee, Gang Woo;Ryu, Ho Jun;Kim, Il Kyung;Seong, Ho;Choi, Chang Hee
    • Pediatric Infection and Vaccine
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    • v.6 no.1
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    • pp.86-92
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    • 1999
  • Purpose : The purpose of this study is to know the epidemiology and clinical characteristics in patients with M. pneumoniae pneumonia during study period retrospectively. Methods : We had reviewed 500 patients with Mycoplasma pneumoniae pneumonia who were hospitalized at the pediatric ward of Seoul Adventist Hospital from July 1986 through June 1996. we had analysed clinical and laboratory problems of pulmonary and extrapulmonary manifestations including demographics and epidemiological characteristics. Results : Peak occurrence of the disease was 3 years interval and seasonal occurrence of the disease had no difference. 43.4% of patients was below 4 years old and male to female ratio was nearly 1:1. The most common site of infiltration on chest PA was right lower lobe and the most common type of pneumonia was interstitial pneumonia(43%). Pulmonary complications were atelectasis(11cases), emphysema(5cases), pleural effusion(27cases) and sinusitis(13cases) and extrapulmonary manifestations were hepatitis(109cases), skin rash(29cases), proteinuria(20cases) and hematuria(16cases). cold agglutinin test was carried out in 500cases and mycoplasma antibody test was carried out in 448cases. sensitivity of mycoplasma antibody test was 53.3%. Conclusion : Mycoplasma pneumoniae pneumonia showed epidemics at intervals of 3 year in its major occurrence. it exhibits a tendency that occurs at younger age group not in school age group. diagnosis of the disease need more accurate method due to low sensitivity of mycoplasma antibody test. also we need to carried out other laboratory test(for example, LFT, U/A).

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Rotavirus P and G Genotypes Circulating in Kyungsangnamdo, Korea, during 2000~2001 (2000~2001년 경상남도에서 유행한 로타바이러스 유전자형)

  • So, Kyeung Jin;Lee, Mi Hyun;Ma, Sang Hyeok;Kim, Byung Chyeol;Yang, Jai Myung
    • Pediatric Infection and Vaccine
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    • v.11 no.1
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    • pp.59-72
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    • 2004
  • Purpose : Rotaviruses are the major cause of gastroenteritis in infants and young children worldwide. It is important to get the epidemiologic data of rotavirus genotype for the application of rotavirus vaccine. So we tried to investigate the distribution of rotavirus genotypes with RT-PCR. Methods : A total of 120 rotavirus latex agglutinin test positive stool samples were collected continually from 120 children from Sep. 2000 to Apr. 2001. Rotavirus P(VP4), G (VP7) genotypes were determined by RT-PCR. Results : The genotype was identified in 116 stool samples of total 120 samples(96%). The incidence of G genotype was as follow; G1 17(14.2%), G2 74(61.7%), G4 1(0.8%), G9 1(0.8%). There were four cases of multiple genotypes; G1/G2, G1/G4, G1/G9, G8/G9 and genotype of G3, G8 were not found. Twenty three(19.2%) samples were nontypeable. The incidence of P was as follow; P[4] 77(64.2%), P[6] 22(18.3%), P4/P6 12(10%), P[4]/P[8] 1(0.8%) p[8] 1(0.8%). Seven(5.9%) samples were nontypeable. Conclusion : Various combinations of G and P genotypes were observed. Most rotavirus strains were P[4]G2 62(51.74%), followed by P[6]G2 7(5.8%), and P[6]G1 7(5.8%), P[4/P[6] G1 4(3%), P[4]/P[6]G2 4(3%), P[4]G1 3(2.5%), P[8]G2 1(0.8%), P[4]G4 1(0.8%) in Kyoungsangnamdo, Korea during 2000~2001.

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ULTRASTRUCTURAL ANALYSIS OF TOOTH PULP AFFERENTS TERMINALS IN THE MEDULLARY DORSAL HORN OF THE RAT (치수유래 구심성 신경섬유의 삼차신경 감각핵군에서의 연접특성)

  • Bae, Yong-Chul;Lee, Eun-Hee;Choy, Min-Ki;Hong, Su-Hyung;Kim, Hyun-Jung;Na, Soon-Hyeun;Kim, Young-Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.2
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    • pp.219-227
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    • 2001
  • Little is known about processing mechanism of pain sensation of the oral cavity at the 1st synapse of trigeminal sensory nuclei. Serial ultrathin sections of tooth pulp afferent terminals, identified by the transganglionic transport of 1% wheatgerm agglutinin conjugated horseradish peroxidase, were investigated with electron microscope. Quantitative ultrastructural analysis was performed on digitizing tablet connected to Macintoshi personal computer (software; NIH Image 1.60, NIH, Bethesda, MD). Labeled boutons could be classified into two types by the shapes of containing vesicles : S bouton, which contained mainly spherical vesicles (Dia. 45-55 nm) and few large dense cored vesicles (Dia, 80-120nm), and LDCV bouton, which contained spherical vesicles as well as large number of large dense cored vesicles. Most of the parameters on the ultrastructural characteristic and synaptic organization of labeled boutons were similar between S and LDCV boutons, except shapes of containing vesicles. Majority of the labeled boutons showed simple synaptic arrangement. The labeled boutons were frequency presynaptic to dendritic spine, and to a lesser extent, dendritic shaft. They rarely synapsed with soma and adjacent proximal dendrite. A small proportion of labeled boutons made synaptic contacts with presynaptic, pleomorphic vesicles containing endings and synaptic triad. Morphometric parameters of labeled boutons including volume and surface area, total apposed area, mitochondrial volume, active zone area, vesicle number and density showed wide variation and these were not significantly different between S and LDCV boutons. The present study revealed characteristic features on ultrastructure and synaptic connection of pulpal afferents which may involved in transmission of oral pain sensation.

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Acceleration of Early Embryonic Development by Induction of Acrosome Reaction in Intracytoplasmic Sperm Injection (세포질내 정자주입술 시행시 정자의 첨체반응이 수정란의 초기 발생과 임신율에 미치는 영향)

  • Lim, Y.J.;Lee, D.R.;Lee, J.E.;Kim, H.J.;Paik, H.R.;Yoon, H.S.;Shim, H.N.;Cho, J.H.;Roh, S.I.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.311-318
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    • 1997
  • Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.

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