• Title/Summary/Keyword: ISH

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Detection of HHV6 and EBV in histiocytic necrotizing lymphadenitis (Histiocytic necrotizing lymphadenitis에서 HHV6과 EBV의 검출)

  • Park, Kyung Hee;Park, Sung Shik;Kim, Ji Yeon;Park, Su Eun
    • Clinical and Experimental Pediatrics
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    • v.51 no.9
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    • pp.987-991
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    • 2008
  • Purpose : Kikuchi-Fujimoto disease (KFD), also known as histiocytic necrotizing lymphadenitis (HNL), is a self-limited disease characterized by cervical lymphadenopathy and fever. The etiology of KFD remains unknown; however, the self-limiting nature of HNL suggests the cause of this disease could be viral infection. For this reason, several viruses have been evaluated as possible etiologies of HNL, including Epstein-Barr virus (EBV), human herpesvirus 6 (HHV6), human herpesvirus 8 (HHV8), and cytomegalovirus (CMV). The aim of this study was to examine the relationship of EBV and HHV6 to HNL. Methods : Data pertaining to 51 cases with biopsy-confirmed HNL were collected between January 1999 and December 2005, from the Department of Pathology, College of Medicine, Pusan National University, Busan, Korea. The clinical records-including data regarding age, gender, duration of fever, and lymph node involvementwere reviewed retrospectively. The in situ hybridization (ISH) assay was performed by EBER PNA probe (Dako, Capinteria, CA, USA), and immunohistochemistry testing was performed with anti-HHV type 6 monoclonal antibodies (Chemicon, Temecula, CA, USA). Results : The HNL patients in this study were 24 males and 27 females, ranging in age from seven to 61 years (median: 25.9). ISH for EBV was positive in 8/51 (15.7%) biopsies, and immunohistochemistry for HHV6 was positive in 15/51 (29.4%) biopsies. Serologic analysis of EBV IgM was performed in 23 cases; only one patient was positive for EBV IgM and EBV ISH. Conclusion : Our study could not provide supportive evidence of a viral pathogenesis for HNL; therefore, cases of HNL may not have a dominant viral cause. However, some rare exceptional cases may have been caused by viral infection.

Detection of EBV-encoded Small RNA from Diffuse Large B-cell Lymphoma Patients by RT-PCR Method

  • Yoo, Tae Hyun;Lee, Min Ho;Park, Min;Lee, Jaewang;Woo, Hyun Jun;Kim, Hyun Woo;Yang, Ji Yeong;Kwon, Hye Jin;Yeon, Min Ji;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.21 no.4
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    • pp.227-232
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    • 2015
  • Epstein-Barr virus (EBV) has a pathogenic role in several lymphomas including diffuse large B-cell lymphoma (DLBCL). In this study, we detected EBV from formalin-fixed paraffin embedded (FFPE) tissues of DLBCL patients by RT-PCR and compared the sensitivity of the RT-PCR method to in situ hybridization (ISH) method. The RNA was extracted from 91 FFPE samples with DLBCL and amplified with primers targeting EBV-encoded small RNA (EBER) by RT-PCR. When using the RT-PCR method, 13 of 91 patients (14.3%) were positive and among these 13 cases, 7 cases (7.7%) were from > 50-year-old patients that is classified as EBV positive DLBCL of the elderly. In previous results using ISH method, 3 of 91 patients (3.3%) were positive and 2 case (2.4%) were older than 50-year-old. These results indicate that RT-PCR method used in this study shows a higher sensitivity than ISH method. The ratio of male versus female among the EBV positive samples was 1.2:1 with the ratio of male higher. If RT-PCR method having high sensitivity is used simultaneously as well as the ISH method providing the information of the EBV positive cellular location, it is expected that EBV will be more accurately detected.

Detection of Gene Amplification by Multiplex Ligation-Dependent Probe Amplification in Comparison with In Situ Hybridization and Immunohistochemistry

  • Tabarestani, Sanaz;Ghaderian, Sayyed Mohammad Hossein;Rezvani, Hamid
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7997-8002
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    • 2015
  • Gene amplification is an important mechanism in the development and progression of cancer. Currently, gene amplification status is generally determined by in situ hybridization (ISH). Multiplex ligation-dependent probe amplification (MLPA) is a PCR-based method that allows copy number detection of up to 50 nucleic acid sequences in one reaction. The aim of the present study was to compare results for HER2, CCND1, MYC and ESR1 gene amplification detected by MLPA with fluorescent in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) as clinically approved methods. Tissue samples of 170 invasive breast cancers were collected. All were ER positive. Tissue samples had previously been tested for HER2 using immunohistochemistry. Amplification of the selected genes were assessed using MLPA, FISH and CISH and results were compared. HER2 MLPA and ISH results were also compared with HER2 immunohistochemistry (IHC) which detects protein overexpression. Amplification of HER2, CCND1, MYC and ESR1 by MLPA were found in 9%, 19%, 20% and 2% of samples, respectively. Amplification of HER2, CCND1, MYC and ESR1 by FISH was noted in 7%, 16%, 16% and 1% of samples, respectively. A high level of concordance was found between MLPA/FISH (HER2: 88%, CCND1: 88%, MYC: 86%, ESR1: 92%) and MLPA/CISH (HER2: 84%). Of all IHC 3+ cases, 91% were amplified by MLPA. In IHC 2+ group, 31% were MLPA amplified. In IHC 1+ group, 2% were MLPA amplified. None of the IHC 0 cases were amplified by MLPA. Our results indicate that there is a good correlation between MLPA, IHC and ISH results. Therefore, MLPA can serve as an alternative to ISH for detection of gene amplification.

Tissue-Specific Localization NUCB2/nesfatin-1 in the Liver and Heart of Mouse Fetus

  • Sun, Sojung;Yang, Hyunwon
    • Development and Reproduction
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    • v.22 no.4
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    • pp.331-339
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    • 2018
  • NUCB2/nesfatin-1 is first known to be expressed in the hypothalamus while controlling appetite and energy metabolism. However, recent studies have shown that NUCB2/nesfatin-1 was expressed in the various organs as well as the hypothalamus. Our previous reports also demonstrated that NUCB2/nesfatin-1 was expressed in the ovary, testis, pituitary gland, lung, kidney, and stomach of fetal and adult mice. However, the role of NUCB2/nesfatin-1 in mouse fetus remains unknown. Thus, the aim of this study was to investigate whether NUCB2/nestatin-1 is expressed in mouse fetus at the developmental stage in which organogenesis begins. To do this, we performed in situ hybridization (ISH) and immunohistochemistry (IHC) staining to examine the distribution of NUCB2 mRNA and nesfatin-1 protein in the mouse fetal organs during early developmental stages, especially at embryonic day (E) 10.5. As a result of ISH, NUCB2 mRNA positive signals were more frequent in the liver, but there were relatively few positive signals in heart. On the other hand, no positive signals were detected in other organs. These ISH results were validated by IHC staining and qRT-PCR analysis. Expression of nesfatin-1 protein detected by IHC staining was similar to that of NUCB2 mRNA detected by ISH in the liver and heart. In addition, the levels of NUCB2 mRNA expression analyzed by qRT-PCR were significantly increased in the liver and heart compared to other organs of the mouse fetus at E13.5, whereas its level was extensively decreased in the liver, but increased in the lung, stomach, and kidney of the mouse fetus at E17.5. These results suggest that NUCB2/nesfatin-1 may play an important role in liver and heart development and physiological functions in the developmental process of mouse fetus. Further studies are needed on the function of NUCB2/nesfatin-1, which is highly expressed in the various organs, including liver and heart during mouse development.

Occupational Health Policies on Risk Assessment in Japan

  • Horie, Seichi
    • Safety and Health at Work
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    • v.1 no.1
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    • pp.19-28
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    • 2010
  • Industrial Safety and Health Law (ISH Law) of Japan requires abnormalities identified in evaluations of worker health and working environments are reported to occupational physicians, and employers are advised of measures to ensure appropriate accommodations in working environments and work procedures. Since the 1980s, notions of a risk assessment and occupational safety and health management system were expected to further prevent industrial accidents. In 2005, ISH Law stipulated workplace risk assessment using the wording "employers shall endeavor." Following the amendment, multiple documents and guidelines for risk assessment for different work procedures were developed. They require ISH Laws to be implemented fully and workplaces to plan and execute measures to reduce risks, ranking them from those addressing potential hazards to those requiring workers to wear protective articles. A governmental survey in 2005 found the performance of risk assessment was 20.4% and common reasons for not implementing risk assessments were lack of adequate personnel or knowledge. ISH Law specifies criminal penalties for both individuals and organizations. Moreover, under the Labor Contract Law promulgated in 2007, employers are obliged to make reasonable efforts to ensure employee health for foreseeable and avoidable risks. Therefore, enterprises neglecting even the non-binding provisions of guidelines are likely to suffer significant business impact if judged to be responsible for industrial accidents or occupational disease. To promote risk assessment, we must strengthen technical, financial, and physical support from public-service organizations, encourage the dissemination of good practices to reduce risks, and consider additional employer incentives, including relaxed mandatory regulations.

A Task Planning System of a Steward Robot with a State Partitioning Technique (상태 분할 기법을 이용한 집사 로봇의 작업 계획 시스템)

  • Kim, Yong-Hwi;Lee, Hyong-Euk;Kim, Heon-Hui;Park, Kwang-Hyun;Bien, Z. Zenn
    • The Journal of Korea Robotics Society
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    • v.3 no.1
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    • pp.23-32
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    • 2008
  • This paper presents a task planning system for a steward robot, which has been developed as an interactive intermediate agent between an end-user and a complex smart home environment called the ISH (Intelligent Sweet Home) at KAIST (Korea Advanced Institute of Science and Technology). The ISH is a large-scale robotic environment with various assistive robots and home appliances for independent living of the elderly and the people with disabilities. In particular, as an approach for achieving human-friendly human-robot interaction, we aim at 'simplification of task commands' by the user. In this sense, a task planning system has been proposed to generate a sequence of actions effectively for coordinating subtasks of the target subsystems from the given high-level task command. Basically, the task planning is performed under the framework of STRIPS (Stanford Research Institute Problem Solver) representation and the split planning method. In addition, we applied a state-partitioning technique to the backward split planning method to reduce computational time. By analyzing the obtained graph, the planning system decomposes an original planning problem into several independent sub-problems, and then, the planning system generates a proper sequence of actions. To show the effectiveness of the proposed system, we deal with a scenario of a planning problem in the ISH.

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Digoxigenin-labeled in situ Hybridization for Diagnosis of Aujeszky's disease in Pigs (Digoxigenin-labeled in situ Hybridization을 이용한 오제스키병의 진단)

  • Kim, Soon-Bok;Moon, Oun-Kyung;Song, Geun-Suk;Yeo, Sang-Geon
    • Korean Journal of Veterinary Pathology
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    • v.1 no.2
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    • pp.119-124
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    • 1997
  • The objective of this study was to develop digoxigenin (DIG)-labeled in situ hybridization (ISH) test for diagnosis of Aujeszky's Disease(AD) in infected organs. Specific DNA with well conserved gene sequences encoding gp50 antigen in AD virus (ADV) was obtained by Polymerase Chain Reaction (PCR) method. A pair of oligonucleotide primers used in PCR allowed amplification of a 217 bp sequence from the gp50 ADV gene. The DNA was then labeled with DIG by primer labeling method for use as probe in ISH test to detect ADV nucleic acids in various tissue. Positive hybridization was demonstrated by dark pigmentation in nuclei and cytoplasm of ADV infected cells particularly in brain tonsillar crypt epithelium and pulmonary alveolar cells. This result suggests that ISH is a valuable sensitive and rapid diagnostic test for AD.

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Histopathological features and viral genome detection in caprine arthritis encephalitis virus infected dairy goats in Korea

  • Son, Gain;Cho, Eun-Sang;Shin, Hyun-Jin;Son, Hwa-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.3
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    • pp.161-168
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    • 2017
  • Caprine arthritis encephalitis (CAE) virus is a causative agent of caprine arthritis-encephalitis. In our previous study we reported a prevalence of CAE. In this study, we described the further detailed pathological features of CAE and examined the detection of virus by in situ hybridization (ISH). Histopathologically, interstitial pneumonia and bronchopneumonia in lung, focal inflammation in mammary glands, perivascular cuffing in brain, arthritis, and focal necrosis, mild steatosis, inflammatory cell infiltration of liver were noted. CAEV proviral-DNA was identified by nested polymerase chain reaction (PCR) in blood cells, brain, synovial fluid, and lymph node. Confirmation by nested PCR involved amplification of a 296 bp ($1^{st}$ PCR) and 185 bp ($2^{nd}$ PCR) fragments corresponding to a conserved region on the gag gene of CAEV. Positive ISH signals were detected in the brain and liver. In conclusion, significant histopathological findings included parenchymal infection in various organs, including the lung, liver, brain, joint, and mammary gland were noted in the CAEV infected dairy goat. ISH can help confirm the diagnosis of CAE in formalin-fixed samples.

Rapid and Easy Diagnosis of Rabbit Haemorrhagic Disease by In Situ Hybridization (In situ Hybridization에 의한 토끼출혈증(rabbit haemorrhagic disease)의 신속.간편한 진단)

  • Park, Nam-Yong;Cho, Ho-Seong;Cho, Kyoung-Oh;Kim, Sang-Jip;Park, Hyung-Seon
    • Korean Journal of Veterinary Pathology
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    • v.5 no.2
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    • pp.57-62
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    • 2001
  • Recently various molecular diagnostic techniques have been used to identify rabbit hemorrhagic disease virus (RHDV), a causative agent responsible for acute hepatitis and disseminated intravascular coagulation in rabbit. But they were hard to perform and time consuming. To detect RHDV in a rapid and easy way, we developed biotinylated oligonucleotide probe within ORF 1 region encoding the polyprotein of RHDV in formalin-fixed and paraffin-embedded tissues from various tissues of 20 rabbits naturally infected with RHDV, Our in situ hybridization (ISH) was quickly carried out within two hours by MicroProbe capillary action system. The ISH produced a positive reaction in liver, kidney and lung. In conclusion, ISH with a biotintlated oligonucleotide probe provided a useful diagnostic method for detecting RHDV.

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