전자현미경적 응용 Shadowing에 의한 핵산 사슬의 구조적 분석

주경웅 ( Kyeng Woong Joo ) 대한임상검사과학회 2003 대한임상검사과학회지(KJCLS) Vol.35 No.2

Rabbit hemorrhagic disease(RHD) which was first recognized in China in 1984. Although Rabbit hemorrhagic disease virus was initially characterized as a picornavirus or a parvovirus, it is now proven to be calicivirus. Kleinschmidt and Zahn developed a method which allows the visualization of DNA molecules as smooth, flexible filaments. In this protein monolayer technique the nucleic acid molecules bind to a protein film which results from surface denaturation of a cytochrome C at an air-water interface. The surface film is then picked up and the nucleic acid molecules are examined in the electron microscope after uranyl acetate staining and heavy metal shadowing. This study is purposed to detection of RHD viral RNA structure by modified shadowing technique as basic procedure of Kleinschmidt and Zahn for observed to difference as ssRNA of RHD viral nucleic acids from other ssDNA on electron microscopy. I have experimented to chemical modified procedures that spreading and diffusion to 90% formamide and 0.2N NaOH denaturing agents, the signal is enhanced by silver solution before electron microscopic observation. It was appeared with effect to dense single strands of RHD viral nucleic acids

면역조직화학염색에서 항원성 회복을 위한 특이 기법에 대한 연구

주경웅 ( Kyeng Woong Joo ) 대한임상검사과학회 2001 대한임상검사과학회지(KJCLS) Vol.33 No.2

Imunohistochemistry is used for antigen localization of estrogen receptor and progesterone receptor in breast tissues. The specificity of the resu1ts depends on two independent criteria : Pathological confmnation of the clinical diagnosis has been a problem because of the difficu1ty in antigen retrieval from formalin fixed, paraffm-embedded specimens. Many different protocols for the detection of antigen in each tissues have been used. πlUS far, proteinasefpepsin, picric anlφ formic acid, hydrated streaming autoclaving at 121 0C of sections, microwave treatment, and 4 M guami이ne thiocyanate treatment have been suggested. The objective of our experiment was to obtain the standard pretreatment resu1ting in optimal immunostaining. In the experiment, successive tissue slides of breast specimens of breast cancer. To study epitope retrieval of antigen using two differential procedure of preπeatments. Using of immunostaining analysis, severe well-defmed locations per section was quantified. The best results were obtained when the heat-induced epitope retrieval method(HIER : hydrated steam Autoclave) is necessary for antigen retrieval 10min steam autoclaving at 121 0C is used, resu1ts in widespread staining of the ER and PR depositin the denated nucleus(DN) and the number of reactive sites and the intensity are significantly reduced, When enzyme-induced epitope retrieval method(EIER) was used, no staining was observed in the DN and on1y faint reactivity was observed in high power lens.

Unicryl 표본에서 광학/전자 현미경 In Situ Hybridization -Rabbit hemorrhagic disease viral RNA 증명-

주경웅 ( Kyeng Woong Joo ) 대한임상검사과학회 1998 대한임상검사과학회지(KJCLS) Vol.30 No.3

Rabbit hemorrhagic disease<RHD) which was first recognized in China in 1984 spread via Eastern Europe to many countries of Western Europe and other parts of the world. This study is purposed. to detection of RHD viral RNA by In situ hybidization(ISH) as the immunohistochemical technique on base of the hemagglutinin inhibition titer and histopathologically changes after inoculation of rabbits with isolates of RHDV obtained from RHD infected cell. After experimental intramuscular infections with isolates of the virus, Infection were leaded acute RHD and showed convulsions, peresis and somtimes bloody or bloody-foamy nasal excretions. The body temperature of all cases show SOIre fever shortly before death. In order to detect antibodies to RlIDV, the hemagglutinin inhibition test. animal which survived an infection with RHDV developed antibodies to RlIDV, which were highly dectable at 12 hours after post infection these antibody titers reached the highest on 48 hours after infection Histopatholgically, in the liver early after infection were detected of hermrrhages. degenerative perilobular alterations in hepatocytes strongly homogenous or pale vacuolized cytoplasm, noclear pyknosis, karyorrhexis of karyolysis were observed Antibodies to RHDV were found in sera that indicate the existence of an pathogenic virus which is related to RHDV Simple stain and light microscopic ISH make possible using semi-thick sections to the post-unicryl embedded block for electron mcroscopic ISH therefore, it was established compared to viral focal location at same region. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probes labelled by biotin cyanoethyl-phosphoramidite compared with 4717-4ro) sequences of RHOV eDNA The best results were obtained to retention and accessibility of RHOV in O.Q20/o glutaraldehyde, depression of aldehyde group by ammonium hydroxide, unicryl resin block and block stain, biotinylated oligonucleotide probes cocktail, acetylation, hybridization during 2 hours at 3TC and signal marked by antibiotin antibody-10nm gold ISH microparticles signal on light microscopic in situ hybridization on semi-thick sections of normal cells and RHOV infected liver using biotinylated probe. It ~ as if indicate the presence of RHO viral nudeic acids by showed gold dense signal, appearing in the cytoplasm of degenerated hepatocytes of sudden death cases. Infected cells are easily recognized to condensed nucleus and central vacuoles, But, The light microscopic ISH were not showed to positive reaction because of limits of resolving power of light microscope and immuno-gold size. At the electron microscopic ISH, RHO viral RNA was distributed widely in the cytoplasm of liver cell. Gold particles were separated of inner and outer nudeanrembrane with destruction of the membrane, and dilatation of rough endoplasmic reticulum in the cell. Many viral RNA gold particles are present in the juxtanuclear cytoplasm and in the vicinity of rER of natural cases of sudden death and dusters of virus particles are seen in the cytoplasm.

쥐 표피세포 순수배양을 위한 초대배양기법에 관한 연구

주경웅 ( Kyeng Woong Joo ) 대한임상검사과학회 2000 대한임상검사과학회지(KJCLS) Vol.32 No.3

This report experimented that to investigate the σypsin treatment for inhibition of fibroblastic contamination and the fetal bovine serum(FBS) effect for trypsin inactivity toprimary culture of mouse foreskin. 0.25 % cold trypsin incubation could removed to dermallayer and subcutaneous materials, and 2.5% FBS σeatment could stopped to trypsin activity. πlose foreskin cells that were maintained in Dulbecco’s Modified Eagle Medium(DMEM) with nonessential amino acid(NEAA) under air conditions equilibrated with 5 % C02, 95 % air and humidified to the water tray. πle inverted microscopic distribution of culture cells at 24 hours after postincubation is showed to c1ear in background at the focus and increased to attachment of c1uster cells at culture well bottom. and then, growth cells and living cell ratio are increased to cold trypsin and FBS treatmented group at 48 hours. Fibroblasts are appeared that some of the spindle-shaped cells derived from dennal lesions to control group, but experimental group is showed to epitheloid cells compare with control group by the silver stain. The trypsin pretreatmented samples showed to epitheloid colonies after postinoculation without other cellul따 contamination to primary culture. πns culture system might provide to a procedure of epidennal cell pure culture.

홍어 장점막 점액의 조직화학적 성상에 관한 연구

주경웅 ( Kyeng Woong Joo ) 대한임상검사과학회 2004 대한임상검사과학회지(KJCLS) Vol.36 No.2

This study is performed in order to clarify the histochemical structure, the distribution of mucous cell and goblet cell, and the histochemical properties of the mucosubstances in the middle region of intestinal mucosa and rectum of Raja kenojei. In the H&E stain, distribution of the mucous cells and acidophilic cells were a more compacted middle portion than other regions of intestine, but the former was more than the latter in the number of mucous cells to rectum. The mucosubstances were stained with aldehyde fuchsin pH 1.7-alcian blue(pH 2.5) stain and then compared to distribution of the mucosubstances used in image and microscope technology(IMT-Size5). The middle intestine of Raja kenojei was composed of mucous cells having only large amounts of mucosubstances in the distal region was much more than that of proximal region. It was two types of mucous cells to rectum, one type was the same as proximal intestine while the other had small amounts of weakly sulfated and large amounts of carboxylated mucins.

2000년 전라남도 지역에서 간접 IFA 에 의한 Orientia tsutsugamushi 항체가 분포 조사

이영종 ( Young Jong Lee ),주경웅 ( Kyeng Woong Joo ) 대한임상검사과학회 2001 대한임상검사과학회지(KJCLS) Vol.33 No.2

To understand the seroepidemiological patterns of scrub typhus in leollanamdo, 199 patients`` sera with acute febrile episodes from each area were tested for the antibody to Orientia tsutsugamushi by indirect immunofluorescent antibody assay technique from May to December in 2000. Out of 199 patients, 173 (86.9%) patients showed positive reaction to 0. tsutsugamushi and the fema1e outnumbered the ma1e. 25.4% of seropositives to o. tsutsugamushi was in the age of the fifties, and 83.2% of seropositives to 0. tsutsugamushi was shown between the forties and seventies of the age. Among each area, 20 showed positive reaction to 0. tsutsugamushi. πlese results might provide the basic information for the management of scrub typhus in Jeollanamdo.

초대배양에서 혈청이 세포증식에 미치는 효과

주경웅 광주보건대학 2000 論文集 Vol.25 No.-

To investigate the fetal bovine serum(FBS) effect for growth and living cells on mouse foreskin cell that this report experimented to primary cluture with serum contain(group Ⅰ) versus serum free(group Ⅱ) culture conditions in the Hank's balanced salt solution(HBSS). Those foreskin cells that were maintained in culture media under air conditions equilibrated with 5% CO, 95% air and humidifyed to the water tray. The inverted microscopic distribution of culture cells at 48 hours after postincubation is that of Group Ⅰ is showed to clear in backgroung at the focus and increased to attachment of unknown cluster cells at culture well bottom. But, growth and living cells appears to increase at group Ⅱ. Fibroblasts are appdared that some of the spindle-shaped cells derived from dermal lesions to group Ⅰ and Ⅱ at 28 hours after postincubation by the silver stain. The FBS was known to growth inhibitors of fibroblast during cell culture, but these serum contained culture system was not inhibited to growth of fibroblasts to compare with serum free media. The results of growth cells have been differented under two culture conditions; the number of epithelial cells of group Ⅰwas showed to increase at only early stage(24 hours), but that of group Ⅱ was increased to compare with group Ⅰ after 48 hours. By these results, It is that of the serum free culture is effected to primary culture for establish of cell line and purification of special constitution of cell.