• Title/Summary/Keyword: staining

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Micronucleus Frequencies in Human Umbilical Cord Blood by the Supravital Staining Method (인체에서의 초생체 염색법을 이용한 제대혈내 소핵 출현 빈도)

  • 박혜경;이은일;류재천;김해준
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.289-295
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    • 2002
  • This study was conducted to quantify of micronucleus frequencies in human umbilical cord blood by supravital staining method with acridine orange, and to find some factors that affected on micronucleus frequncies in humans. In this study, we used umbilical cord blood of new born infants that have sufficient reticulocytes compared with adult peripheral blood. The cord bloods were taken after childbirth from 60 normal infants in industrial and coastal region in Korea. The total of 3 ${mu}ell$ cord blood was applied to slide coated with acridine orange, and micronuclei were observed under fluorescent microscopy. Demographic factors and independent variables were collected from mothers by questionnaire. The frequencies of micronuclei in umbilical cord blood of new born infants were 0-5 per 2,000 reticulocytes by supravital staining method, and mean value and standard deviation were 1.75$\pm$0.97. There were no significant difference by the regions, smoking habits of father or mother. However, age of mother showed significant positive correlation with frequencies of micronuclei (p<0.05). Smoking at home by fathers also was found as a significant variable by muliple regression analysis. Therefore, further studies would be needed for genotoxicological evaluation of new born infants by microneuli test using supravital staining method.

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Animal Skin Pigmentation Model Using Full Thickness Skin Graft in C57BL/6 Mouse (C57BL/6 마우스의 등에 시행한 자가 전층피부이식편을 이용한 색소침착 동물모델)

  • Lee, Hong-Ki;Park, Jong-Lim;Heo, Eun-Ju;Kim, Suk-Wha
    • Archives of Plastic Surgery
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    • v.38 no.6
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    • pp.725-732
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    • 2011
  • Purpose: Skin grafting is one of the most commonly used methods in reconstructive plastic surgery field, but complications such as color change, contracture or hypertrophy are common problems. However, pathophysiology of the color change after skin graft is not yet determined and no animal model is established. Methods: Full thickness skin grafts were performed on the dorsum of C57BL/6 mice. Serial chronological gross inspection for color change and pigmentation were examined. Melanin pigments were traced by Fontana-Masson staining and semi-quantitative analysis was performed. In addition, immunohistochemical staining of S-100, Micropthalmia related Transcription Factor (MITF) and Melan-A antibodies were also performed to observe melanocytes and their changes. Results: After skin graft, color change and pigment spots were observed in the graft. Fontana-Masson staining showed melanin pigments in the epidermal and dermal layers in all mice. Immunohistochemistry staining to S-100, MITF, Melan-A antibodies showed melanocytes at the basal layer of epidermis and dermis. Conclusion: In conclusion, we have established an animal model for skin pigmentation after skin graft. We believe this study may be useful in understanding of the behavior of melanocytes after skin graft.

Detection of Mycobacterium tuberculosis by PCR from Trace Clinical Specimens and Paraffin-embedded Tissues (임상가검물과 파라핀 포매 조직에서 PCR법을 이용한 결핵균의 검출)

  • 김은중;최우순;황석연
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.55-63
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    • 2000
  • This study has been carried out to investigate the sensitivity of polymerase chain reaction (PCR) method over conventional acid-fast bacilli (AFB) staining and/culture methods for the detection of Mycobacterium tuberculosis from trace body fluid and paraffin-embedded tissues (PET) specimens. A total of 65 cases were employed for the AFB staining and culture test, and a total of 50 cases were subjected to PCR and histopathological analysis. Among the specimen showing negative reaction to AFB staining, 12.1% were positive to PCR and 3.7% of the specimen representing negative result to culture test showed positive reaction to PCR. In addition, 20.0% of the specimen with AFB negative showed positive reaction to PCR. From these results, it could be concluded that PCR method overwhelms AFB staining and culture tests in sensitivity and specificity to M tuberculosis detection.

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Changes in the Neurogenesis and Axonal Sprouting in the Organotypic Hippocampal Slice Culture by Aβ25-35 Treatment

  • Jung, Yeon Joo;Jiang, Hui Ling;Lee, Kyung Eun
    • Applied Microscopy
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    • v.42 no.4
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    • pp.200-206
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    • 2012
  • Induction of neurogenesis can occur in the hippocampus in response to various pathological conditions, such as Alzheimer's disease. The aim of this study was to investigate the changes that occur in endogenous neural stem cells in response to amyloid beta $(A{\beta})_{25-35}$-induced neuronal cell damage in organotypic hippocampal slice cultures. Cresyl violet staining and Fluoro-Jade B staining were used to detect neuronal cell damage and changes of mossy fiber terminals were observed by Timm's staining. The immunofl uorescence staining was used to detect the newly generated cells in the subgranular zone (SGZ) of the dentate gyrus with specific marker, 5-bromo-2'-deoxyuridine (BrdU), Ki-67, Nestin, and doublecortin (DCX). In compared to control slices, neuronal cell damage was observed and the mossy fibers were expanded to CA3 area by treatment with $A{\beta}_{25-35}$. Ki-67/Nestin- and BrdU/DCX-positive cells were detected in the SGZ. In conclusion, these results demonstrate that $A{\beta}$-induced neuronal damage results in an increase in endogenous neural stem cells in rat hippocampal slice cultures not only for gliosis but also for neurogenesis.

New record of five Euplotes species(Protozoa, Ciliophora) collected from South Korea

  • Jeong Hyeon Yeo;Pablo Quintela-Alonso;Jae-Ho Jung
    • Journal of Species Research
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    • v.12 no.3
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    • pp.203-211
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    • 2023
  • Five ciliate species of Euplotes were isolated from fresh and coastal water during a sampling survey to identify unrecorded ciliates in South Korea. Their morphology was investigated using live observation, protargol and "wet" silver nitrate staining methods. Brief descriptions and microphotographs of each species and a comparison with related species are provided. Euplotes focardii is characterized by an average size of 65×47 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-eurystomus type. Euplotes nobilii shows an average size of 34×20 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes octocarinatus, the only freshwater species described in the present study, is characterized by an average size of 66×46 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes petzi has an average size of 43×30 ㎛ after protargol staining, a macronucleus hook-shaped and dorsal argyrome pattern in double-patella type. Euplotes raikovi is characterized by an average size of 40×24 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type.

Protective effect of platelet-rich plasma against cold ischemia-induced apoptosis of canine adipose-derived mesenchymal stem cells

  • Suji Shin;Sung-Eon Kim;Seong-Won An;Seong-Mok Jeong;Young-Sam Kwon
    • Korean Journal of Veterinary Research
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    • v.64 no.1
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    • pp.2.1-2.8
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    • 2024
  • This study was performed to assess the antiapoptotic effect of canine platelet-rich plasma (PRP) treated on the canine adipose-derived mesenchymal stem cells (cMSCs) under cold ischemic conditions. The effect of preventing apoptosis of cMSCs was evaluated in the apoptotic condition induced by cold ischemic injury in vitro. To determine the progression of apoptosis, the changes in cell nucleus were observed using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining. In addition, we examined the mitochondrial membrane potential (MMP) and caspase-3 activity. When the cold hypoxic injury was applied to cMSCs, the apoptotic change was observed by DAPI staining, mitochondrial staining for MMP, and caspase-3 assay. PRP significantly decreased the number of apoptotic cells. Nuclear shrinkage and fragmentation of apoptotic cells in control groups were observed by DAPI staining. The MMP was recovered by the treatment of PRP. In addition, when the luminescence intensity was measured for caspase-3 activity, the value was significantly higher in the PRP treated groups than the control groups. The results of this study showed that the PRP may have a beneficial effect on apoptosis induced by cold ischemic injury.

Studies on the Colorfastness to Perspiration of Knitted Blouse (편직물 Blouse의 (땀)에 대한 염색견로성에 대한 연구)

  • 이원자
    • Journal of the Korean Home Economics Association
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    • v.11 no.3
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    • pp.285-298
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    • 1973
  • The colorfastness of dying persipiration and laundry on summer clothing must be considered, because it has special relation to the human body. The colors of fibers as cotton, p/c, acryl, polyester and nylon which have been widely used for blouse and T-Shirt of Knitted wear are R-P, Y-G, BI-B and print. Studies were carried out with persipirometer, for the natural fiber of cotton the chemical one of nylon, with additional stuff involved, which polluted. The experiment was conducted to colorfastness with acid solution and alkaline solution to see the alteration of color and staining of man-made persiperation. The results obtained from this experiment can be summerized as follows. 1. The order of color alteration isnylon < p/c < coton < polyester < acryl, and the nylon shows the lowest colorfastness, which is 3 class, and the acrly shows the highest colorfateness, which is 5 class. The staining of multifiber test of cotton fabric is nylon < p/c < polyester < cotton < acryl. The staining of multifiber of nylon fabric is nylon polyester < p/c < cotton < acryl. 2. In acid solution and alkaline solution, the alteration of color and staining makes almost no difference, but concerning staining of cotton, the acid solution is lower than the case of alteration solution only. 3. In the pollution on cotton and nylon, the latter is more easily polluted than the former regardless of fabrics. Especially in case of polluted nylon, ti shows the lowest color fastness (2 class), which causes a problem of the dying process and dye stuffs. 4. No difference of color alteration shows among them, but R-P and print show low color fastness (2 class), especially printed nylon shows the lowest value (1 class).

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Follow Up Expression Patterns of Alkaline Phosphatase(AP) as a Marker for Establishing Mouse Embryonic Stem (ES) Cells (배아주간세포수립을 위한 Alkaline Phosphatase(AP)의 상이한 발현 양식의 추적)

  • 김진회;차수경;노민경;송상진;구덕본;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.55-63
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    • 1995
  • The putative totipotency germ cells has a relative abundance of alkaline phosphatases. Thus, histological staining of AP activity offers a new route to isolate totipotent cells and also provides insights into culture systems of these cells. Furthermore, the AP staining technique is simple and fast, requires only the napthol AS/MS substrate in combination with trapping diazonium salts such as fast red or fast blue. However, our unexpected finding was that AP staining of mouse ES cells were detected in the undifferentiaed epiblast-derived cells as well as several types of differentiating cells. This findings are different from results of Talbot et al. (1993) reported usefulness of the AP staining and implies that histological staining of AP may not by useful to determine undifferentiaed state or totipotency of ES cells. Thus, we have investigated the patterns of AP expression by RT-PCR in order to identify a marker of undifferentiated ES/primordial germ (PG) cells. In RT-PCR analysis, embryonic (E)-AP was detected only in undifferentiated ES cells, but intestinal(I)-AP was not detected in all of the examined ES and PG cells. In addition, nonspecific (NS)-AP wasdetected in undifferentiated PG cell from day 7, 5 to 13 of gestation. Histological activity of AP in ES cells was completely suppressed by addition of L-phenylalanine (Phe), L-homoarginine (Har), and L-phenylalanylglycylglycine (PheGlyGly) as an inhibitor, but RT-PCR showed the same results as in the absence of an inhibitors. Our findings suggested that expression of E-AP and NS-AP may use as a marker to determine the undifferentiated status in ES and PG cells.

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Comparisons of C-kit, DOG1, CD34, PKC-θ and PDGFR-α Expressions in Gastrointestinal Stromal Tumors According to Histopathological Risk Classification

  • Kim, Ki-Sung;Song, Hye-Jung;Shin, Won-Sub;Song, Kang-Won
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.2
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    • pp.48-56
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    • 2011
  • Gastrointestinal stromal tumor (GIST) is a mesenchymal tumor and is associated with a specific immunophenotype index. It is very important to identify the specific immunophenotype and the diagnosis for the treatment GIST patients. Ninety two cases of GIST analyzed in this study were immuno-stained for c-kit, DOG1, CD34, PKC-${\theta}$, PDGFR-${\alpha}$. The rate of positive staining and statistical significance were then compared. In addition, the GISTs were analyzed as followings: very low risk, low risk, intermediate risk and high risk according to tumor size and nuclear division, and later correlated with clinical parameters. The results of the GIST positive stainings were: DOG1 (95.7%), PKC-${\theta}$ (90.2%), PDGFR-${\alpha}$ (88.0%), c-kit (87.0%) and CD34 (71.7%). Only DOG1 staining showed a statistical significance of p<0.05. It was identified in the classification system of histologic risk that staining expression of DOG1, PKC-${\theta}$, PDGFR-${\alpha}$ were significantly increased as histologic risk increases (p<0.05). However, clinical parameters such as age and sex of patients have no correlations with the classification system of histologic risk (p>0.05). Therefore, in this study, the expression of DOG1 showed statistical significance and DOG1, PKC-${\theta}$, PDGFR-${\alpha}$ staining increased significantly as the histologic risk increases in histologic classification system. Taken together, the DOG1 staining should be very effective for the diagnosis of GIST patients.

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Opisthorchis viverrini Infection Activates the PI3K/AKT/PTEN and Wnt/β-catenin Signaling Pathways in a Cholangiocarcinogenesis Model

  • Yothaisong, Supak;Thanee, Malinee;Namwat, Nisana;Yongvanit, Puangrat;Boonmars, Thidarut;Puapairoj, Anucha;Loilome, Watcharin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10463-10468
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    • 2015
  • Opisthorchis viverrini (Ov) infection is the major etiological factor for cholangiocarcinoma (CCA), especially in northeast Thailand. We have previously reported significant involvement of PI3K/AKT/PTEN and $Wnt/{\beta}$-catenin in human CCA tissues. The present study, therefore, examined the expression and activation of PI3K/AKT/PTEN and $Wnt/{\beta}$-catenin signaling components during Ov-induced cholangiocarcinogenesis in a hamster animal model. Hamsters were divided into two groups; non-treated and Ov plus NDMA treated. The results of immunohistochemical staining showed an upregulation of PI3K/AKT signaling as determined by elevated expression of the $p85{\alpha}$-regulatory and $p110{\alpha}$-catalytic subunits of PI3K as well as increased expression and activation of AKT during cholangiocarcinogenesis. Interestingly, the staining intensity of activated AKT (p-AKT) increased in the apical regions of the bile ducts and strong staining was detected where the liver fluke resides. Moreover, PTEN, a negative regulator of PI3K/AKT, was suppressed by decreased expression and increased phosphorylation during cholangiocarcinogenesis. We also detected upregulation of $Wnt/{\beta}$-catenin signaling as determined by increased positive staining of Wnt3, Wnt3a, Wnt5a, Wnt7b and ${\beta}$-catenin, corresponded with the period of cholangiocarcinogenesis. Furthermore, nuclear staining of ${\beta}$-catenin was observed in CCA tissues. Our results suggest the liver fluke infection causes chronic inflammatory conditions which lead to upregulation of the PI3K/AKT and $Wnt/{\beta}$-catenin signaling pathways which may drive CCA carcinogenesis. These results provide useful information for drug development, prevention and treatment of CCA.