• Title/Summary/Keyword: antibody formation

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Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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Studies on Host-Virus Interaction of Poxviruses 1. Cytochemical, Autoradiographic and Immunocytological Analysis in Cowpox Virus-FL Cell System (Poxvirus 감염(感染)에 있어서의 Virus-숙주세포(宿主細胞)의 상호관계(相互關係) 1. Cowpox Virus-FL 세포계(細胞系)의 세포화학적(細胞化學的) Autoradiography 및 세포면역학적해석(細胞免疫學的解析))

  • Kim, Uh Ho
    • Korean Journal of Veterinary Research
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    • v.15 no.1
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    • pp.57-67
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    • 1975
  • The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

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Effect of Full-Fat Flax, $\alpha$-Tocopherol and Selenium on Phagocytes Chemiluminescence of Broiler Chickens (아마종실과 $\alpha$-Tocopherol, 셀레늄 급여가 육계 혈액의 탐식세포로부터 생성되는 $O_{2}$-와 $H_{2}$ $O_{2}$ 에 의해 유도된 화학발광(chemiluminescence) 생성)

  • 안종남;채현석;김동운;권명상;박병성
    • Korean Journal of Poultry Science
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    • v.28 no.3
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    • pp.239-244
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    • 2001
  • To examine the effects of feed additives on the expression of Perpheral blood cell surface molecules, phagocytosis and antigen specific antibody formation, broilers were randomly assigned to T$_1$, T$_2$, T$_3$, and T$_4$ groups. T$_1$ group was fed diet without any additives for 13 weeks, T$_2$ was fed diet with full fat flax, T$_3$ was fed diet with full fat flax containing $\alpha$-tocopherol, and T$_4$ was fed diet with full-fat flax containing $\alpha$-tocopherol and selenium. Since 5 weeks feeding the data were examined by luminometer. After 2 weeks adminstration of different feeding, although all treated groups (T$_2$, T$_3$, and T$_4$,) showed slightly increased chemiluminescence (CL) responses than T$_1$, this result was not significant. After 4 weeks feeding there was no significant increase of CL in the Phagocytes like neutrophils and macrophages of T$_2$ group compared to T$_1$. But phagocytes from T$_3$ and T$_4$ group showed in creased $O_2$- (6%, 18% respectively) as well as $H_2O$$_2$ (9.5% and 10.9%, respectively) induced CL responses. After 8 weeks feeding there was more than 50% increase $O_2$- induced CL in T$_3$ and T$_4$ group, but $H_2O$$_2$ induced CL responses in T$_3$ and T$_4$ group was slightly increased (6.6% and 9.3%, respectively).

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A Research on Perception and Exposure to Hepatitis-B Virus of Dental Hygienists (치과위생사의 B형 간염에 대한 인지도와 노출에 대한 조사)

  • Kang, Eun-Ju;Kim, Jin;Choi, Mi-Hye
    • Journal of dental hygiene science
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    • v.7 no.1
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    • pp.13-20
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    • 2007
  • This study is aimed at providing basic data for measures to prevent dental hygienists who are highly exposed to hepatitis-B virus due to their characteristic working conditions from being infected of it. In order to determine their perception and the extent of their exposure to the risk of infection with hepatitis-B virus, a survey was conducted with questionnaires distributed to 354 dental hygienists who are working at dental offices in Seoul. Gyeongi and Jeolla provincial areas. From the survey, following conclusions could be drawn: 1. Sixty three point seven to 100 percent of them are found to have been vaccinated against hepatitis-B virus as is generally expected from their working environment. 2. Their general characters are found to be statistically significant in terms of vaccinating point of time among those who have been vaccinated. Most respondents are either vaccinated one year or 5 years ago. The completion of 3 requested vaccinations lies in the level of 64.3%~100% depending on the general characters. By age, the best result comes from the age group of 25~29, followed by the groups older than 30 and 20-24 in order (p < 0.05). Formation of antibody belongs to the level of 45%~100% generally. But statistically significant of them is the extremely unhealthy state with 100%, followed by normal, generally healthy, very healthy and not healthy in sequence (p < 0.05). 3. In terms of family's clinical history, it is known that the older (p < 0.01), the more unhealthy (p < 0.01) and the married (p < 0.01), the more possibility of having anamnesis of hepatitis-B virus infection. 4. The level of knowledge about Hepatitis-B virus is shown to be in the extent of 4.39~5.01 out of maximum 8 points in general terms. 5. It is revealed that there are high chances for dental hygienists to get spattered with body matters of patients like blood or sputum on their faces in general characters (87.%~100%). 6. A high rate (75%~100%) of respondents has experienced being shot by needles. The older(p < 0.05) and the longer(p < 0.05) their career, the higher the rate becomes.

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The changed expression of CGRP immune-positive nerve in the periodontal ligament of rat molar during tooth movement (실험적 치아이동에 의한 백서 치주인대내 CGRP 면역 양성 신경섬유의 변화)

  • Sung, Jung-Ok;Park, Hyo-Sang;Bae, Yong-Chul;Sung, Jae-Hyun
    • The korean journal of orthodontics
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    • v.26 no.5 s.58
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    • pp.581-590
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    • 1996
  • Bone resorptiion was predominate in compression site, bone formation in tension site of periodontal ligament during tooth movement. The biologic response at compressiion site was different from tension site. Thus the CGHP immuno-positive nerve fiber will respond differently to mechanical force according to the area( compression or tension site). The purpose of this study was to investigate this response of CGRP immune-positive nerve fiber in the periodontal ligament according to the duration of applied force and the area (compression or tension site) during experimental tooth movement. The experimental animals were 7 week old male rat (approximately 200 gm). The orthodontic force was applied mesially to the right maxillary molar using the Ni-Ti coil spring during 12hours, 1, 3, 7, and 120days. Immunohistochemical staining using antibody against CGRP was performed after sacrifice. The results were as follows. The CGRP immune-positive nerve bundle showed reduced immunoreactivity and nerve fibers reduced in density after application of orthodontic force for 12 hours and 1day. The CGRP immune-positive nerve fibers showed many thin branches at the apical periodontal ligament after application of force for 3 days as compared with normal. The tension site in the apical periodontal ligament showed more branches than the compression site. In 7 day group, the CGRP immune-positive nerve fibers increased in terms of the number and had many thin branches in the apical periodontal ligament. The tension site had more branches than the compression site. In 12 day group, the CGRP immune-positive nerve fibers showed similar distribution to normal control at compression site of apical periodontal ligament, but the fibers at the tension site increased in number. The CGRP immuno-positive nerve fibers showed more increased at tension site than compression site after application of orthodontic force. Therefore it seems to have some relation to the bone remodeling besides the local inflammatory process.

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A Study on the Knowledge and Awareness of Dental Hygienists and Dental Hygiene Students about Hepatitis B (치과위생사와 치위생과 학생의 B형 간염에 대한 지식과 인식에 관한 조사연구)

  • Kim, Jeong-Suk;Choi, MI-Hye;Kang, Eun-Ju
    • Journal of dental hygiene science
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    • v.8 no.4
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    • pp.233-239
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    • 2008
  • The purpose of this study was to survey the knowledge and awareness of dental hygienists and dental hygiene students about hepatitis B in an attempt to lay the solid foundation for the prevention of hepatitis B and infection control, as dental hygienists were highly likely to be exposed to HBV during job performance. The findings of the study were as follows: 1. 32.5% of the dental hygienists and 15.6% of the dental hygiene students were aware of their own HBsAg and HBsAb states. The dental hygienists who were cognizant of the states statistically significantly outnumbered the dental hygiene students who were (p=.000). 2. As for vaccination against hepatitis B, 71.8% of the dental hygienists and 47.3 % of the dental hygiene students were inoculated against it. The dental hygienists who were vaccinated against it outnumbered the dental hygiene students who were, and the disparity between the two was statistically significant (p=.000). 3. In the event of those who were vaccinated against hepatitis B, the dental hygienists were better cognizant than the dental hygiene students of the right time for the vaccination, required vaccination frequency (three times) and confirmation of the formation of hepatitis B antibody (p=.000). 4. Regarding awareness of hepatitis B infection route, the dental hygienists knew significantly better than the dental hygiene students that hepatitis B might be infected via blood (p=.030), sexual relations (p=.000), contaminated needle sticks (p=.000), mothers with hepatitis B positive during delivery (p=.000), toothbrushes/razors (p=.000) and exchange of drinking cups (p=.000). 5. As to the relationship between health status and knowledge on hepatitis B infection route, the respondents who were in bad shape had the best knowledge about that, followed by those in an average state of health and healthy respondents. And the dental hygienists had a significantly better knowledge than the dental hygiene students (p=.001). 6. Just a small number of the dental hygienists and students knew about the hepatitis B-related past experiences of their families, and the gap between the two was insignificant.

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Generation of a Mammalian Gene Expression Vector Using Bovine Viral Diarrhea Virus (Bovine Vira1 Diarrhea Virus를 이용한 포유동물세포 발현벡터의 개발)

  • 이영민
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.86-95
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    • 2002
  • As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.