• Title/Summary/Keyword: Immunocytochemical staining

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Immunocytochemical Staining for p16 of Atypical Squamous Cells in Cervicovaginal Smear (자궁목질 바른표본에서 비정형 편평세포의 p16에 대한 면역세포화학염색)

  • Kim, Hwal-Woong;Lee, Jong-Sil;Lee, Jeong-Hee;Ko, Gyung-Hyuck
    • The Korean Journal of Cytopathology
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    • v.15 no.1
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    • pp.28-32
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    • 2004
  • It was reported that the main cause of intraepithelial neoplasm and squamous cell carcinoma of the uterine cervix is human papilloma virus infection, and that the expression of p16 is increased in cells infected by human papilloma virus. We performed an immunocytochemical staining for protein p16 in 17 cases of cervocovaginal smears initially diagnosed as atypical squamous cells of undetermined significance, to know whether the staining could help the differentiation of neoplastic cells from reactive atypical cells. Of 17 smears, 6 were diagnosed finally as high grade intraepithelial neoplasm or invasive squamous cell carcinoma by follow-up biopsy and smear, and 5 of the 6 were positive for p16. Three were diagnosed as koilocytosis, and one of them was weakly positive for p16. Eight were diagnosed as reactive atypical cells, and all of them were negative for p16. We thought that immunocytochemical staining of p16 in cervocovaginal smears could help the differentiation of neoplastic cells from reactive atypical cells.

Usefulness of p16INK4a Immunocytochemical staining for the Differentiation between Atrophy and ASCUS in Diagnosis of Uterine Cervical Cancer

  • Hye Ryoung Shin;Taekil Eom;Wan-Su Choi
    • Biomedical Science Letters
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    • v.29 no.3
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    • pp.144-151
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    • 2023
  • A Pap smear is the most important screening test for the diagnosis of cervical cancer. However, subjective judgment by the operator cannot be excluded, and replicability may greatly be reduced if uncertain specimens are examined. Examiners often experience difficulties in differentiating atrophy with inflammatory changes and ASCUS when diagnosing squamous epithelial lesions from a pap smear. Reports often vary between cytologists and pathologists, and misdiagnosis may result in delayed follow-ups and advanced diseases. Hence, auxiliary examinations are necessary when confusing results between atrophy and ASCUS are obtained. The importance of p16INK4a activation due to HPV infection, which is an important factor in the outbreak of cervical cancer, has been highlighted. Recent studies have reported that p16INK4a immunocytochemical staining and HPV high-risk type tests using liquid-based cervical specimens are effective to detect the presence of lesions of grade HSIL or higher in patients with ASC-H. However, no research exists on the utility of HPV and p16INK4a tests on the differential diagnosis of atrophy and ASCUS. This study focused on whether p16INK4a immunocytochemical staining and HPV tests can help diagnose borderline lesions between atrophy and ASCUS. The results reported that p16INK4a activation can significantly (P<0.001) differentiate atrophy from ASCUS in atrophic lesions infected with High risk-HPV. Therefore, it may be concluded that p16INK4a immunocytochemical staining is an effective auxiliary test in lesions infected with HR-HPV when atrophic lesions are difficult to differentiate by morphology. Such results are expected to help decide on adequate follow-up and treatment.

Immunocytochemical Detection of Pneumocystis carinii in Bronchoalveolar Lavage (기관지 폐포 세정액에서 뉴우모시스티스 카리니의 면역세포화학적 검출)

  • Kwon, Kun-Young;Cho, Seung-Che;Kim, Sang-Pyo;Park, Kwan-Kyu;Chang, Eun-Sook;Kim, Chung-Sook
    • The Korean Journal of Cytopathology
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    • v.8 no.1
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    • pp.27-34
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    • 1997
  • Pneumocystis carinli is an established cause of pulmonary infections in immuno-compromised hosts. Several cytoiogical stains, such as Papanicolaou, Gomori methenamine sliver(GMS) and Diff-Quik have been used for detection of the organism, but occasionally can be laborious and, due to a degree of nonspecificity, may be misleading. We evaluated the diagnostic utility of immunocytochenmical stains that recognize P. carinii in bornchoalveolar lavage from experimentally Induced P. carinii pneumonia rats(n=15). In audition to routine stains for diagnosis by morphologic recognition of P. carinii on Papanicolaou, GMS and Diff-Quik stains, bronchoalveolar lavage samples were reacted with immunocytochemical stains using monoclonal antibodies(MAB) 092 and 902. In bronchoalveolar lavage P. carinii organisms were detected In 9 of 10 cases(90%) using each MAB 092 and 902, whereas GMS and Diff-Quik stains demonstrated P. carinii in 13(86%) and 11(73%) of 15 cases respectively. In lung tissue specimens(n=15) P. carinii organisms were well identified on GMS stain and immunohistochemical stains using MAB 092 and 902 in ail cases. We believe that the immunocytochemical staining using MAB 092 and/or 902 is a very useful and diagnostic tool In addition to GMS and Diff-Qulk stain to detect P. carinii organisms in bronchoalveolar lavage.

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Localization of Cyclin B and Erk1/2 in Ovine Oocytes and MPF and MAPK Activities in Cytoplast and Karyoplast following Enucleation

  • Lee, Joon-Hee;Campbell, Keith H.S.
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.407-414
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    • 2011
  • The development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is dependent upon numerous factors. Central to development is the quality and developmental competence of the recipient cytoplast and the type of the donor nucleus. Typically metaphase of the second meiotic division (MII) has become the cytoplast of choice. Production of a cytoplast requires removal of the recipient genetic material, however, it may remove proteins which are essential for development or reduce the levels of cytoplasmic proteins to influence subsequent reprogramming of the donor nucleus. In this study, enucleation at MII did not affect the activities of either MPF or MAPK kinases. Immunocytochemical staining showed that both Cyclin B1 (MPF) and Erk1/2 (MAPK) were associated with the meiotic spindle of AI/TI oocytes with little staining in the cytoplasm, however, at MII association of both proteins with the spindle had reduced and a greater degree of cytoplasmic distribution was observed. The analysis of oocyte proteins removed during enucleation is a difficult approach to the identification of factors which may be depleted in the cytoplast. This is primarily due to the large numbers of aspirated karyoplasts which would be required for the analysis.

Improvement of Neuronal Differentiation by PDE4 Inhibition in Human Bone Marrow-mesenchymal Stem Cells (인간 골수유래-중간엽 줄기세포(hBM-MSCs)에서 PDE4 억제조절을 통한 신경세포 분화 효율 개선)

  • Jeong, Da Hee;Joe, I-Seul;Cho, Goang-Won
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1355-1359
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    • 2016
  • Human bone marrow mesenchymal stem cells (hBM-MSCs) can differentiate into various cell types including osteoblasts, adipocytes, chondrocytes, and myocytes. Previous studies, including our own, have shown that MSCs can also differentiate into neuron-like cells. However, their rate of neuronal differentiation is not sufficient for application to stem cell therapy, which requires well-defined cell types. For this purpose, we first examined the expression of neuronal lineage markers (GFAP, MAP-2, KCNH1, Nestin, NF-M, and Tuj-1) by real-time PCR, western blot, and immunocytochemical staining. The expressions of the astrocyte marker GFAP and neuronal markers NF-M and Tuj-1 increased in neuronal differentiated MSCs (dMSCs). To improve the neuronal differentiation efficiency, PDE4, an important signaling intermediator in the progression of neuronal differentiation, was modulated using well-known inhibitors such as rolipram or resveratrol and then differentiated into neuronal cells (Roli- or RSV-dMSCs). The expressions of NF-M, Tuj-1 were increased while that of GFAP decreased in Roli- and RSV-dMSCs, which were examined by real-time PCR, western blot, and immunocytochemical staining. From these experiments, we have found that the neuronal differentiation efficiency can be ameliorated by the modulation of PDE4 activity.

Composite Epithelioid Hemangioendothelioma in Pleural Effusion Minicking Metastatic Adenocarcinoma - Cytologic and Immunocytochemical Findings - (늑막삼출액에서 전이성 샘암종과 유사한 복합 상피성 혈관내피종 - 세포학적 및 면역세포화학적 소견 -)

  • Jang, Ki-Seok;Han, Hong-Xiu;Park, Moon-Hyang
    • The Korean Journal of Cytopathology
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    • v.14 no.1
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    • pp.36-41
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    • 2003
  • Epithelioid hemangioendothelioma is a rare vascular tumor of borderline malignancy which is characterized by the presence of "epithelioid" or "histiocytoid" endothelial cells. Superficial and deep tumors have been recognized in the extremities, head, neck, chest, and mediastinum of adult patients. It may also occur as a primary tumor of liver, bone, and other visceral organs. Few effusion cytologic findings of epithelioid hemangioendothelioma have been reported. We report a case of composite epithelioid hemangioendothelioma with focal epithelioid angiosarcomatous areas of the iliac bone and adjacent soft tissue in a 38-year-old female, which, during its metastatic course, was presented as a pleural effusion. The effusion was cellular with epithelioid cells presenting both singly and in clusters. The tumor cells were round to ovoid shewing cytoplasmic vacuolization, variability in cell size, and prominent nucleoli. The effusion smears and cell block sections revealed strong positive staining for CD31 and vimentin, weak positive for CD34 and Factor VIII-related antigen, and negative for cytokeratin, CEA, and calretinin. The cytologic findings in this case were similar to that of metastatic adenocarcinoma or malignant mesothelioma. Therefore, immunocytochemical staining in smear and cell block is a helpful tool to differentiate malignant 'epithelioid' cells in effusion.

The Principles and Practices of Immunocytochemical method in Light Microscopic Level (광학 현미경적 수준에서의 면역조직화학적 방법의 원리 및 실제)

  • Kim Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.3 no.1
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    • pp.229-250
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    • 1991
  • The study was carried out to investigate and review the principles and practices of immunocytochemical method in light microscopic level. The results were as follows. 1. Immunocytochemistry is the method to search out the intracellular position of the specific meterials using antigen -antibody reaction. 2. The chief items in immunocytochemistry are antigen, antibody and chromogen. 3. The identifical fixation is cardiac perfusion fixation. 4. The tissue slides must be prepared by vibratomy. 5. All stainings are carried out with free floating staining method. 6. There are polyclonal and monoclonal antibodies used in immunocytochemistry.

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Colocalization of ${\alpha}$of Gq Protein with Actin Filaments in L8E63 Cells

  • Chae, Sungsuk;Park, Dongeun
    • Animal cells and systems
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    • v.1 no.1
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    • pp.93-98
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    • 1997
  • The present study investigated the cellular localization of a-subunit of Gq (Gaq) protein in developing L8E63, rat skeletal muscle cell line. The colocalization of Gaq with actin cytoskeleton was demonstrated by double-labeling experiments. In mononucleated myoblasts, the immuno-fluorescence staining pattern of Gaq was almost identical with that of F-actin visualized with rhodamine-conjugated phalloidin. However, this colocalization of Gaq with cytoskeleton was not maintained in multinucleated myotubes. The staining pattern of Gaq in myotubes did not match with any specific subcellular structure, but appeared as a uniformly distributed diffuse staining throughout the whole cell surface. Interestingly, change in the expression level of Gaq was not detected during myoblast differentiation, suggesting that actin-associated Gaq protein might dissociate from the cytoskeleton as cells differentiate. Immunocytochemical experiments using specific antibodies directed against several G proteins indicated that the subcellular localizations of Gai1, Gai2, Gai3, and Gao were different from those obtained with Gaq.

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Possibility of Undifferentiated Human Thigh Adipose Stem Cells Differentiating into Functional Hepatocytes

  • Lee, Jong Hoon;Lee, Kuk Han;Kim, Min Ho;Kim, Jun Pyo;Lee, Seung Jae;Yoon, Jinah
    • Archives of Plastic Surgery
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    • v.39 no.6
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    • pp.593-599
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    • 2012
  • Background This study aimed to investigate the possibility of isolating mesenchymal stem cells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stem cells (HTASCs) to differentiate into hepatocytes. Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue. Growth factors, cytokines, and hormones were added to the collagen coated dishes to induce the undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm the experimental results, the expression of hepatocyte-specific markers on undifferentiated and differentiated HTASCs was analyzed using reverse transcription polymerase chain reaction and immunocytochemical staining. Differentiation efficiency was evaluated using functional tests such as periodic acid schiff (PAS) staining and detection of the albumin secretion level using enzyme-linked immunosorbent assay (ELISA). Results The majority of the undifferentiated HTASCs were changed into a more polygonal shape showing tight interactions between the cells. The differentiated HTASCs up-regulated mRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intensely stained with anti-albumin antibody compared with undifferentiated HTASCs. PAS staining showed that HTASCs submitted to the hepatocyte differentiation protocol were able to more specifically store glycogen than undifferentiated HTASCs, displaying a purple color in the cytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCs could secrete albumin, which is one of the hepatocyte markers. Conclusions MSCs were islolated from human thigh adipose tissue differentiate to heapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but also thigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.

Utility of Calretinin in Distinction between Benign Reactive Mesothelial and Carcinoma Cells in Serous Effusions (전액성 삼출액내 반응성 중피세포와 암종세포간의 감별진단에서 calretinin의 유용성)

  • Kim, Byung-Heon
    • The Korean Journal of Cytopathology
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    • v.12 no.2
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    • pp.89-95
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    • 2001
  • The cytological distinction of carcinoma cells from reactive mesothelial cells in serous effusions nay be difficult or imposslble based on morphology alone, especially In specimens containing reactive mesothelial cells which form glandular or ball- or papillary-shaped conglomerates or which mimic malignant nuclear features. Calretinin is a newly reported immunocytochemical marker for mesothelial cells, which can potentially be utilized for facilitating this distinction. This study evaluated the usefulness of calretinin for the discrimination between reactive mesothelial and metastatic carcinoma cells in serous effusion. Immunocytochemical staining was undertaken on 33 benign reactive and 87 malignant serous effusion specimens with histologically confirmed diagnoses. The specimens including smears and cell blocks were stained with polyclonal antibody to calretinin by labelled streptavidin-biotin method. The positive expression of calretinin was noted In 32(97.0%) of 33 benign reactive effusions and 9(10.3%) of 87 malignant effusions. The sensitivity and specificity of the calretinin immunostaining for reactive mesothelial cells was 97.0% and 89.7%, respectively. In conclusion, calretinin is a useful marker for distinguishing between reactive mesothelial cells and carcinoma cells in serous effusions.

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