• Title/Summary/Keyword: Negative staining

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Molecular Structure of Muscle Filaments Determined by Electron Microscopy

  • Craig, Roger
    • Applied Microscopy
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    • v.47 no.4
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    • pp.226-232
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    • 2017
  • Electron microscopy and X-ray diffraction have together played a key role in our understanding of the molecular structure and mechanism of contraction of muscle. This review highlights the role of electron microscopy, from early insights into thick and thin filament structure by negative staining, to studies of single myosin molecule structure, and finally to recent high-resolution structures by cryo-electron microscopy. Muscle filaments are designed for movement. Their labile structures thus present challenges to obtaining near-atomic detail, which are also discussed.

Evaluation of the Genotoxicity of Cadmium Chloride in Mice Using the Micronucleus Test

  • Kalantari, Heybatullah;Akhbari, Arash;Elliott, Simon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.255-258
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    • 2002
  • In order to determine the safety of chemicals and pharmaceutical products, various methods can be used to evaluate the toxicity. In this study the genotoxic effect of the widely used industrial chemical, cadmium chloride, was assessed using the micronucleus test in peripheral blood of mice. The presence of micronucleated reticulocytes by microscopic observation following acridine orange staining indicated a potential genotoxic effect. The genotoxicity of intraperitoneally (i.p.) administered cadmium chloride (0.5, 1, 2 mg/kg) appeared to be dose dependent, with the maximum tolerated dose (MTD) found to be 2 mg/kg. Compared to the negative control (saline), cadmium chloride (2 mg/kg) exhibited statistically significant genotoxic potential (P<0.05) but was found to be less than the positive control of mitomycin C (0.5 mg/kg) and was not statistically significant compared to historical negative controls (P>0.05).

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Characterization of Tobacco rattle virus(TRV-K) isolated in Korea (한국에서 분리한 Tobacco rattle virus(TRV-K)의 특성)

  • Shin, Hye-Houng;Koo, Bong-Jin;Kang, Sang-Gu;Chang, Moo-Ung;Ryu, Ki-Hyung
    • Research in Plant Disease
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    • v.8 no.4
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    • pp.207-214
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    • 2002
  • Tobacco rattle virus(TRV) was detected from Gladiolus hybridus, Crocus spp. and Narcissus spp. leaves show-ing notched or stripe on the leaf and malformation symptoms collected from Daegu and Kyungbuk province by electron microscopy (EM), immunosorbent electron microscopy (ISEM) and host range study. Direct negative staining method by EM showed rigid rod long particles 170~200$\times$22 nm and rigid rod short particles 40~114$\times$22 m. TRV-K isolated from G. hybridus propagated with Nicotiana tabacum. TRV coat protein(CP) gene was amplified using specific oligonucleotide primer by RT-PCR. Sequence analysis of amplified CP gene showed 99.5% nucleotide similarity to TRV-ORY.

Somatostatin Receptor 2 and 5 Expressions in Gastroenteropancreatic Neuroendocrine Tumors in Turkey

  • Yerci, Omer;Sehitoglu, Ibrahim;Ugras, Nesrin;Cubukcu, Erdem;Yuce, Suleyman;Bedir, Recep;Cure, Erkan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.10
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    • pp.4377-4381
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    • 2015
  • Background: Gastroenteropancreatic neuroendocrine tumors (GNs) are slow growing and although their incidence has increased in recent years, they are relatively rarely seen. Somatostatin analogues are used in the treatment of GNs that express somatostatin receptor (SR). We aimed to investigate the expression of SR2 and SR5 in GNs. Materials and Methods: In this study the expression of SR2 and SR5 was investigated immunohistochemically in 49 cases (26 males, 23 females) diagnosed and graded with GN according to the World Health Organization classification 2010. Results: The percentage of SR2 staining was 91.0% in grade 1, 82.8% in grade 2 and 100% in grade 3. On the other hand, the percentage of SR5 staining was 81.8% % in grade 1, 60.0% in grade 2 and 0% in grade 3. According to the tumor localization, the percentages of SR2 expression were as follows: pancreas 85.7%, stomach 100%, small bowel 70%, appendix 85.7% and rectum 100%. The percentages of SR5 expression were: pancreas 61,9%, stomach 37.5%, small bowel 70%, appendix 71.5% and rectum 66.6%. There was a significant negative correlation between ki67 percentage and SR5 expression (r=-0.341, p=0.016). Conclusions: In this study, GNs were found to highly express SR2 and SR5. Although the expression of SR2 and SR5 changed according to tumor localization, the expression of SR2 was higher than the expression of SR5 in GN. There was a significant negative correlation between ki67 and SR5. Accordingly, SR5 may be a prognostic indicator of GN.

Isolation and Identification of Influenza Virus from Pusan in 1998 (1998년도 부산지역에서의 Influenza 바이러스의 분리)

  • 조경순;김영희
    • Journal of Life Science
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    • v.9 no.3
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    • pp.289-292
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    • 1999
  • This study was performed to monitor the circulation of various influenza virus strains since influenza is one of the commonest respiratory disease in man, its causative virus has been the subjects of extensive research. The authors investigated the epidemics of influenza in Pusan in 1998. Influenza viruses have been isolated from patients with respiratory disease whose ages range from 1 to 68. Virus isolation from female was higher than male. The isolation of virus was mostly concentrated in December in 1998. The isolated virus showed strong cytopathic effect on MDCK cells and identified as influenza A/Sydney/05/97-like(H3N2) and influenza A/Beijing/262/95-like(H1N1). A negative staining of electron micrograph showed 130 nm with H1N1 in diameter, respectively.

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Domperidone Exerts Antitumor Activity in Triple-Negative Breast Cancer Cells by Modulating Reactive Oxygen Species and JAK/STAT3 Signaling

  • Rajina Shakya;Mi Ran Byun;Sang Hoon Joo;Kyung-Soo Chun;Joon-Seok Choi
    • Biomolecules & Therapeutics
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    • v.31 no.6
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    • pp.692-699
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    • 2023
  • The lack of molecular targets hampers the treatment of triple-negative breast cancer (TNBC). In this study, we determined the cytotoxicity of domperidone, a dopamine D2 receptor (DRD2) antagonist in human TNBC BT-549 and CAL-51 cells. Domperidone inhibited cell growth in a dose- and time-dependent manner. The annexin V/propidium iodide staining showed that domperidone induced apoptosis. The domperidone-induced apoptosis was accompanied by the generation of mitochondrial superoxide and the down-regulation of cyclins and CDKs. The apoptotic effect of domperidone on TNBC cells was prevented by pre-treatment with Mito-TEMPO, a mitochondria-specific antioxidant. The prevention of apoptosis with Mito-TEMPO even at concentrations as low as 100 nM, implies that the generation of mitochondrial ROS mediated the domperidone-induced apoptosis. Immunoblot analysis showed that domperidone-induced apoptosis occurred through the down-regulation of the phosphorylation of JAK2 and STAT3. Moreover, domperidone downregulated the levels of D2-like dopamine receptors including DRD2, regardless of their mRNA levels. Our results support further development of DRD2 antagonists as potential therapeutic strategy treating TNBC.

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.

Diagnostic Utility of p63 (Ab-1) and (Ab-4) Tumor Markers in the Squamous Cell Carcinomas of Head and Neck

  • Khan, Nauman Rauf;Khan, Amna Nauman;Bashir, Saira;Khan, Ayyaz Ali;Suleman, Bilquis A.;Chaudhry, Saima
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.975-978
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    • 2012
  • P63 is a gene product required in cell cycle regulation which plays vital roles in tumor differentiation. Aims of the present study were to assess the frequency, pattern, sensitivity and specificity of two p63 protein clones P63 4A4 and P63 4A4+Y4A3 in squamous cell carcinomas (SCCs). Thirty cases of head and neck region SCC diagnosed on the basis of H&E staining were examined along with 60 cases of head and neck region biopsies other than squamous cell carcinoma, negative on H&E staining, were taken as control. Immunostaining was performed on slides according to the Thermo Scientific UltraVision LP detection System. P63 4A4+Y4A3 clone is more sensitive 96.6% in comparison to 86% in P63 4A4 with having greater NPV of 98.3%. The results signify the importance of P63 4A4+Y4A3 marker over the old markers and may be used as a confirmatory marker of squamous cell carcinoma.

Septic arthritis of the hip joint caused by Klebsiella pneumoniae: a case report

  • Jeong-Bo Moon;Jun-Hwan Lee;Byung-Ju Ryu
    • Journal of Yeungnam Medical Science
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    • v.40 no.2
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    • pp.193-197
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    • 2023
  • Klebsiella pneumoniae is an uncommon cause of septic arthritis in adults. However, late detection can cause serious complications, including joint destruction and immobility. The purpose of this study was to report a case of successfully treated septic arthritis of the hip joint (SAHJ) caused by K. pneumoniae. A 49-year-old female patient presented to our hospital with fever and progressive severe pain in the right hip area. Although there was no abnormality on plain radiographs, ultrasonography revealed diffuse swelling of the right hip joint. Under ultrasonography guidance, the hip joint fluid was aspirated, and Gram staining and culturing were performed. The patient's pain was significantly reduced after the joint aspiration. The Gram staining and culturing revealed gram-negative bacilli, which were subsequently identified as K. pneumoniae. According to the results, systemic intravenous antibiotic (ceftriaxone) was administered without complications, and the patient was discharged on oral antibiotic (ciprofloxacin). Clinical cases of septic arthritis of the knee or sacroiliac joint have been occasionally reported in adults, but cases of SAHJ are rare. Moreover, K. pneumonia-induced SAHJ has not been reported to date. Therefore, we report this very rare case and its successful treatment.

The Polymerase Chain Reaction in Diagnosis of Small B-Cell Non-Hodgkin Lymphomas

  • Antoro, Ester Lianawati;Dwianingsih, Ery Kus;Indrawati, Indrawati;Triningsih, FX Ediati;Harijadi, Harijadi
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.491-495
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    • 2016
  • Background: Small B-cell non-Hodgkins lymphoma (NHL) is difficult to be distinguished from non-neoplastic reactive processes using conventional haematoxylin-eosin (HE) staining due to different interpretations among pathologists with diagnosis based on morphologic features. Ancillary examinations such as immunohistochemical (IHC) staining are essential. However, negative or doubtful results are still sometimes obtained due to unsatisfactory tissue processing or IHC technique. The polymerase chain reaction (PCR) as a molecular diagnostic technique is very sensitive and specific. Clonality detection of heavy chain immunoglobulin (IgH) gene rearrangement has been widely used to establish diagnosis of B-cell NHL. Aims: To elaborate interobserver variation in small B-cell NHL diagnosis based on morphologic features only and to confirm sensitivity and specificity of the PCR technique as an ancillary method. Materials and Methods: A toptal of 28 samples of small B cell NHL and suspicious lymphoma were interpreted by 3 pathologists in Sardjito General Hospital based on their morphology only. The reliability of assessment and the coefficient of interobserver agreement were calculated by Fleiss kappa statistics. Interpretation results were confirmed with IHC staining (CD20, CD3, Bcl2). PCR was performed to analyze the clonality of IgH gene rearrangement. Results: Interobserver agreement in morphologic evalution of small B cell NHL and chronic lymphadenitis revealed kappa coefficient 0.69 included in the substantial agreement category. The cases were divided into 3 groups based on morphology and IHC results; lymphoma, reactive process and undetermined group. PCR analysis showed 90% sensitivity and 60% specificity. Conclusions: The present study revealed a substantial agreement among pathologists in small B-cell NHL diagnosis. For difficult cases, PCR is useful as complementary method to morphologic and IHC examinations to establish definitive diagnosis.