• Title/Summary/Keyword: Negative stain

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An Electron Microscopic Structure of Rotavirus by Negative Stain (Negative stain을 이용한 Rotavirus의 투과전자현미경적 구조)

  • Kwon, Jung-Kyun
    • Applied Microscopy
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    • v.24 no.4
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    • pp.107-114
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    • 1994
  • The Rotaviruses are members of the family Reoviridae and are the major cause of severe childhood gastroenteritis worldwide. Recently, electron microscopy has been used to detect non-group A rotaviruses to determine a relatively high resolution structure of the rotavirion. Mature, infectious virions(double-shelled particles) have a diameter of approximately 70nm, and have a capsid structure composed of two concentric protein layers. We have studied patient's stool specimen by negative staining technique complete removal of sucrose suspension. This negative staining technique that could be carried out in about 30 minutes and that could be used with crude stool specimen was an advantage of major significance. Removal of sucrose in the sample by has been completed washing with distilled of sucrose and by washing with distilled water. Ultrastructurally, typical feature of rotavirus has a double capsid construction with an inner capsid of 55nm and on outer 65-70nm diameter can be clearly demonstrated.

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The Gram-Stain Characteristics of the Bacterial Community as a Function of the Dynamics of Organic Debris in a Hypereutrophic Lake (과 부영양형 호수의 유기물 변동에 따른 박테리아 군집의 그램 염색 특성)

  • Kang, Hun
    • 한국해양학회지
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    • v.24 no.3
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    • pp.148-156
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    • 1989
  • This investigation was performed in eutrophic lake within the framework of a series of studies to evaluate the significance of gram reaction for both bacterioplankton and attached bacteria in the dynamics of organic materials at various aquatic ecosystems. In Lake Kasumigaura as a representative of the highly eutrophic freshwater environments, the gram-stain characteristics of the bacterial community changed with the influx of pulses of phytoplankton, as those in the meso trophic environments. The predominency of the gram-negative forms in the bacterial community was about 57% for bacterioplankton and about 53% for attached bacteria. The statistical analysis of the difference of these two distributions showed that these communites were different. Both gram-negative and gram-positive bacteria attached to particles were shown to effect the formation and degradation of particulate organic matter. Gram-negative bacteria plankton participate exclusively in the dynamics of dissolved organic matter.

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Usefulness of Real-time PCR to Detect Mycobacterium tuberculosis and Nontuberculous Mycobacteria (결핵균과 비결핵성항산균 검출에 Real-time PCR의 유용성)

  • Yun, Eun-Young;Cho, Su-Hee;Go, Se-Il;Baek, Jong-Ha;Kim, You-Eun;Ma, Jeong-Eun;Lee, Gi-Dong;Cho, Yu-Ji;Jeong, Yi-Yeong;Kim, Ho-Cheol;Lee, Jong-Deok;Kim, Sun-Joo;Hwang, Young-Sil
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.4
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    • pp.250-255
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    • 2010
  • Background: The purpose of this study was to evaluate recently developed real-time polymerase chain reaction (PCR) assay kit to detect Mycobacterium tuberculosis (MTB) and nontuberculous mycobacteria (NTM) in respiratory specimens. Methods: We assessed the positive rate of the real-time PCR assay to detect MTB and NTM in 87 culture-positive specimens (37 sputum, 50 bronchial washing), which were performed real-time PCR by using $Real-Q_{TM}$ MTB&NTM Kit from January 2009 to June 2009, at Gyeongsang University Hospital. To compare the efficacy with the TB-PCR assay, we evaluated 63 culture-positive specimens (19 sputum, 44 bronchial washing) for MTB or NTM, which were performed TB-PCR by using ABSOLUTETM MTB II PCR Kit from March 2008 to August 2008. Results: Among 87 specimens tested using real-time PCR, MTB and NTM were cultured in 58 and 29, respectively. The positive rate of real-time PCR assay to detect MTB was 71% (22/31) and 92.6% (25/27) in AFB stain-negative and stain-positive specimens. For NTM, the positive rate of real-time PCR was 11.1% (2/18) and 72.7% (8/11) in AFB stain-negative and stain-positive specimens. Among 63 specimens performed using TB-PCR, MTB and NTM were cultured in 46 and 17, respectively. The positive rate of TB-PCR was 61.7% (21/34) and 100% (12/12) in AFB stain-negative and stain-positive specimens. TB-PCR was negative in all NTM-cultured 17 specimens. Conclusion: TB/NTM real-time PCR assay is useful to differentiate MTB and NTM in AFB stain-positive respiratory specimens and it is as effective in detecting MTB with TB-PCR.

High-Resolution CT Findings of Active Pulmonary Tuberculosis : Different Features Between AFB Stain Positive and Negative Group (활동성 폐결핵의 HRCT 소견 : 객담 도말 양성군과 음성군간의 비교)

  • An, Jeon-Ok;Yoon, Bo-Ra;Jung, Jin-Young;Kim, Yoo-Kyung;Baek, Man-Sun;Kim, Ki-Up;Na, Moon-Jun
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.5
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    • pp.709-719
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    • 2000
  • Background : The different features of high-resolution CT(HRCT) findings of active pulmonary tuberculosis(TB) were studied between acid fast bacilli(AFB) smear or culture positive and negative group. Methods : We prospectively evaluated 36 patients who had been confirmed for active pulmonary tuberculosis by the smear or culture of AFB in sputum(n=25), and changes on serial chest radiographs(n=11). The patients were divided into 3 groups by the results of sputum AFB stain and culture. Group 1(n= 11) is negative in both AFB stain and culture; group 2(n=13) is negative in AFB stain but positive in culture ; and group 3(n=12) is positive in both AFB stain and culture. We evaluated the findings of HRCT in each group randomly. Result : On the HRCT scans, acinar nodule(100%), macronodule(75%), and cavity(75%) in group 3 were more frequently found than group 1(63%. 18%, 9%) and group 2(46%, 15%, 23%)(p<0.05). The centrilobular nodule and branching structure were more frequently observed in group 3(92%) than in group 1(54%)(p<0.05), but were similarly observed in group 2(77%)(p>0.05). AFB positive group was statistically different than the negative group in the HRCT findings with to acinar nodule(100% vs 54%), macronodule(75% vs 17%), and cavity(75% vs 17%)(p<0.05). TB culture positive group was statistically different than the negative group in the HRCT findings with respect to acinar nodule(72% vs 45%) and cavity(48% vs 9%)(p<0.05). Conclusions : HRCT scans are helpful in determining disease acitivity in sputum AFB stain-negative pulmonary tuberculosis. When HRCT shows centrilobular nodule and branching structure, acinar nodule, macronodule, cavity, further studies as sputum induction and bronchoscopy can be performed to determine the presence of bacilli in patients of AFB stain-negative tuberculosis.

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Differentiation of mixed bacterial populations by modified gram stain (수정된 Gram 염색법에 의한 혼합세균 개체군의 분별 측정)

  • 장진경;임종락;정계효;한홍의
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.244-248
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    • 1987
  • Attempts were made to enumerate the number of Gram positive and negative bacteria in the development of natural fermentation rapidly and simultaneously. A general Gram stain was applied to this study. The number of cells by Gram stain was proportional to the cell turbidity by spectrophotometer within a range of 0.7 absorbance at 610nm. The cells washed out during procedures were not exceeded about 8 percentage. The standard error of separate counts in the mixture of Cscherichia coli and Micrococcus luteus was $5.1\pm2.3$%. The possible range of counting was $5.5\times 10^{7}-1.0\times 10^{9}$ cells/ml. Therefore, it is believed that a general Gram stain could be applied to the separate counting of mixture of Fram positive and negative bacterial populations too. In practice, growth kinetics of hemp retting and Kimchi fermentation were presented.

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Different Profiles of the Negatively Stained Citrus Canker Bacterium Xanthomonas citri pv. citri Depending on Culture Media and Heavy Metal Stains

  • Kim, Ki-Woo;Lee, In-Jung;Hyun, Jae-Wook;Lee, Yong-Hoon;Park, Eun-Woo
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.90-92
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    • 2010
  • Staining profiles and bacterial morphology were compared in Xanthomonas citri pv. citri by a transmission electron microscopy. Four types of negative staining regimes were employed depending on culture media and heavy metal stains. The bacterial cells grown on LB agar media often appeared clustered on the supporting film. Meanwhile, individual bacterial cells could be readily found on the preparations from LB broth media. Typical rod-shaped cells (ca. $1\;{\mu}m$ in length) and their flagella were observed in either 2% uranyl acetate (UA) or 2% neutralized potassium phosphotungstate (PTA) staining. The UA-stained bacteria often showed relatively intact cell morphology and rather positively stained cells with a thin electron-dense stain depth around bacteria. The PTA-stained bacteria were characterized by the wrinkled cell surface where the stain was entrapped in grooves. In addition, distinct electron-dense stain depth was evident around the PTA-stained preparations. Numerous fimbriae could be mostly observed from the PTA-stained preparations of the two culture media, but not from the UA-stained preparations.

Spontaneous intaocular amelanotic melanomas in ktc:Sprague-Dawley rats (Ktc:Sprague-Dawley 랫드의 안구에 자연발생한 amelanotic melanoma)

  • Kang, Boo-hyon;Son, Hwa-young;Ha, Chang-su;Kim, Dae-yong
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.375-381
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    • 1995
  • Two intraocular tumors were identified in low and medium dosed groups of a carcinogenicity study using 200 males and 200 females Sprague-Dawley rats in Screening & Toxicology Research Center, Korea Research Institute of Chemical Technology. The tumors were grossly observed as white or yellow, unilateral nodules. They were approximately $1-2{\times}3-5mm$ in size. The tumors located in the region of iris and/or ciliary body invaded peripheral cornea. The microscopic features were usually composed of spindle cells arranged in parallel, forming gently curving bundles or whorls. The spindle cells had poorly defined cell boundaries, scant to moderate cytoplasm, fusiform nuclei and indistinct nucleoli. Mitotic figures were rare and areas of necrosis were present. The spindle cells had positive immunoreactivity for S-100 protein and vimentin but negative for desmin, collagen and HMB-45 antibody. In special histochemical studies, the spindle cells react with Gomori's stain for argyrophih fibers, Prussian blue stain for iron but negative with Masson-Fontana's stain for melanin granules. Ultrastructurally, cytoplasmic premelanosomes were not observed in the tumor cells due to the poor preservation of tumor masses. Based on the results, the tumors were diagnosed as amelanotic melanoma.

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Effect of Washing Solution Characteristics on the Removal and Color of Cocoa Stains (세탁용수의 특성에 따른 코코아 오구의 세척성과 색상)

  • Chung, Hae-Won;Kim, Hyo-Jeong
    • Fashion & Textile Research Journal
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    • v.14 no.3
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    • pp.492-500
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    • 2012
  • Cocoa is a popular drink for children and contains healthy polyphenols however; a deep brown stain is left when cocoa is spilled over clothes. The main pigments in cocoa are anthocyanins that change in washing solutions with different alkalinity and metals. The removal and color changes in a cocoa stain after washing with various pH solutions and water hardness were studied. Alkalinity and the water hardness of washing solutions were important factors for the removal of cocoa stains. The removal of cocoa things in washing solutions without detergent was low (and even became negative after removal and darker) in solutions with a pH 9 and above. The cocoa stain was not removed and only the fabric color faded, although the cocoa stained cloth was washed with Korea tap water that has a pH of 7. The cocoa stain removal in detergent solutions was conspicuously higher than for only water. Even in detergent solutions, the cocoa stain removal decreased as water hardness increased. Cocoa stain removal was more effective and the color dimmest when the stained cloth was washed in a solution without the metal cations, and the bleach added with the detergent at a temperature of $40^{\circ}C$ and for longer than 20 minutes. Effective and economical equipment for tap water softening for a washing machine should be developed and used to improve cocoa stain removal.

Performance of Quantitative Real-Time PCR for Detection of Tuberculosis in Granulomatous Lymphadenitis Using Formalin-Fixed Paraffin-Embedded Tissue

  • Munkhdelger, Jijgee;Mia-Jan, Khalilullah;Lee, Dongsup;Park, Sangjung;Kim, Sunghyun;Choi, Yeonim;Wang, Hye-Young;Jeon, Bo-Young;Lee, Hyeyoung;Park, Kwang Hwa
    • Biomedical Science Letters
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    • v.19 no.2
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    • pp.153-157
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    • 2013
  • Although culture is the gold standard method to identify mycobacteria, its use in tuberculous lymphadenitis (TBL) is limited due to formalin fixation of the submitted specimens. We evaluated the performance of quantitative real-time PCR (q-PCR) for Mycobacterium Tuberculosis (MTB) in granulomatous lymphadenitis using formalin-fixed paraffin-embedded (FFPE) tissues. From 2000 to 2010, a total number of 117 cases of lymph node samples with granulomatous inflammation which were surgically removed and fixed in formalin were studied. Hematoxylin & Eosin (H&E) and Ziehl-Neelsen-stained (ZN) slides were reviewed. qPCR using Real TB-Taq$^{(R)}$ was performed for all cases to identify Mycobacterium tuberculosis. Thirteen non-tuberculous lymphadenopathy cases were used as negative control. Cervical lymph nodes were more frequently affected (60%, 70/117) than other sites. ZN stain for acid fast bacilli was positive in 19 (16.24%) cases. qPCR for tuberculosis was positive in 92 (78.63%) cases. Caseous necrosis was found in 103 (88.03%) cases. While the ZN stain and qPCR were both negative in all control cases, the qPCR showed a significantly higher positive rate (78.63% vs. 16.24%) compared to ZN stain in histologically diagnosed TBL. Quantitative real-time PCR proves to be more sensitive than ZN stain for diagnosis of tuberculous lymphadenitis.

IMMUNOHISTOCHEMICAL STUDY OF TGF-β ON THE HEALING PROCESS IN THE RAT FEMUR BONE DEFECT (백서의 골 결손부 치유시 Transforming Growth Factor-β의 발현에 대한 면역 조직 화학 연구)

  • Bang, Sung-Jun;Kim, Kyung-Wook;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.20 no.2
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    • pp.173-183
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    • 1998
  • $TGF-{\beta}$ is one of growth factors that may be involved in the formation of bone and cartilage. Multiple studies demonstrate that $TGF-{\beta}$ is involved in regulating cell proliferation, differentiation and matrix synthesis, events observed in frature healing. The apperance of $TGF-{\beta}$ in the fracture during healing was evaluated by immunohistologic localization of $TGF-{\beta}$ using antibody. Twenty Sprauge-Dawley strain white male rats, each weiging about 150grams were used and divided two groups. The one group, the $2{\times}2mm$ bony defect was formed in the right femur. The other group, $4{\times}2mm$ bony defect was formed in right femur. Both group were sacrificed at 3day, 1, 2, 3, 4 week and femur were harvested, paraffin sections were stained with H & E, MT stain, immunihistochemical staining with $TGF-{\beta}$ antibody and observed under light microscope. The result were as follows: 1. New bone formation and cartilagenous tissue was seen at 3day. And in the $2{\times}2mm$ bony defect group, $TGF-{\beta}$ stained the cell surounding new bone. 2. The osteoclast and trabecular was seen at 1week. $TGF-{\beta}$ stained the osteoblast and in the $2{\times}2mm$ bony defect group was stained more than $4{\times}2mm$ bony defect group. 3. The lamella bone and trabecule was seen from 3, 4week, and $TGF-{\beta}$ stained almost negative. From the above finding, we could concluded that $TGF-{\beta}$ stained the osteoblast at early stage and 1week, the peak stain was seen from 1week, and then decreased, almost negative stain was seen at 3, 4week.

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