• Title/Summary/Keyword: In-situ diagnosis

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Immunocytochemistry, In situ hybridization and electron microscopy for early diagnosis of Aujeszky's in living pigs (오제스키병의 생체 조기진단을 위한 면역세포화학, In situ hybridization 및 전자현미경적 연구)

  • Moon, Oun-kyong;Kim, Soon-bok;Sur, Jung-hyang;Song, Geun-suk;Nho, Whan-gook
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.845-858
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    • 1996
  • The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

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A Case of Successful Pregnancy in Patient with Recurrent Spontaneous Abortion by Preimplantation Genetic Diagnosis Following IVF-ET (염색체 이상에 의한 반복 유산 환자에서 체외수정시술 및 착상전 유전진단을 통한 임신 성공 1례)

  • Jeong, Jin-Seok;Yeon, Gyu-Sun;Chae, Hee-Dong;Cheon, Yong-Pil;Kim, Chung-Hoon;Kang, Byung-Moon;Chang, Yoon-Seok;Mok, Jung-Eun
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.135-140
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    • 1998
  • It was reported that the etiologies of recurrent spontaneous abortion are immunologic factors, endocrinologic problems, anatomical abnormalities, genetic abnormalities, infection, and unexplained factors. Among those etiologic factors, genetic abnormalities occur in about 5% of the couples who experience recurrent spontaneous abortions, and most common parental chromosomal abnormality contributing to recurrent abortion is balanced translocation. The advent of in vitro fertilization (IVF), the development of skills associated with the handling of human embryo, and an explosion of knowledge in molecular biology have opened the possibility of early diagnosis of genetic disease in preimplantation embryos. Therefore preimplantation genetic diagnosis (PGD) is indicated for couples, infertile or not, at risk of transmitting a genetic disease. A case of successful pregnancy and term delivery by PGD using fluorescence in situ hybridization (FISH) technique in patient with recurrent spontaneous abortion due to balanced translocation is presented with brief review of literatures.

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Fractal dimension analysis as an easy computational approach to improve breast cancer histopathological diagnosis

  • Lucas Glaucio da Silva;Waleska Rayanne Sizinia da Silva Monteiro;Tiago Medeiros de Aguiar Moreira;Maria Aparecida Esteves Rabelo;Emílio Augusto Campos Pereira de Assis;Gustavo Torres de Souza
    • Applied Microscopy
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    • v.51
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    • pp.6.1-6.9
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    • 2021
  • Histopathology is a well-established standard diagnosis employed for the majority of malignancies, including breast cancer. Nevertheless, despite training and standardization, it is considered operator-dependent and errors are still a concern. Fractal dimension analysis is a computational image processing technique that allows assessing the degree of complexity in patterns. We aimed here at providing a robust and easily attainable method for introducing computer-assisted techniques to histopathology laboratories. Slides from two databases were used: A) Breast Cancer Histopathological; and B) Grand Challenge on Breast Cancer Histology. Set A contained 2480 images from 24 patients with benign alterations, and 5429 images from 58 patients with breast cancer. Set B comprised 100 images of each type: normal tissue, benign alterations, in situ carcinoma, and invasive carcinoma. All images were analyzed with the FracLac algorithm in the ImageJ computational environment to yield the box count fractal dimension (Db) results. Images on set A on 40x magnification were statistically different (p = 0.0003), whereas images on 400x did not present differences in their means. On set B, the mean Db values presented promising statistical differences when comparing. Normal and/or benign images to in situ and/or invasive carcinoma (all p < 0.0001). Interestingly, there was no difference when comparing normal tissue to benign alterations. These data corroborate with previous work in which fractal analysis allowed differentiating malignancies. Computer-aided diagnosis algorithms may beneficiate from using Db data; specific Db cut-off values may yield ~ 99% specificity in diagnosing breast cancer. Furthermore, the fact that it allows assessing tissue complexity, this tool may be used to understand the progression of the histological alterations in cancer.

Prenatal diagnosis of an unbalanced translocation between chromosome Y and chromosome 15 in a female fetus

  • Lee, Dongsook;Park, Heeju;Kwak, Sanha;Lee, Soomin;Go, Sanghee;Park, Sohyun;Jo, Sukyung;Kim, Kichul;Lee, Seunggwan;Hwang, Doyeong
    • Journal of Genetic Medicine
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    • v.13 no.2
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    • pp.95-98
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    • 2016
  • We report the prenatal diagnosis of an unbalanced translocation between chromosome Y and chromosome 15 in a female fetus. Cytogenetic analysis of parental chromosomes revealed that the mother had a normal 46,XX karyotype, whereas the father exhibited a 46,XY,der(15)t(Y;15) karyotype. We performed cytogenetic analysis of the father's family as a result of the father and confirmed the same karyotype in his mother and brother. Fluorescence in situ hybridization and quantitative fluorescent-polymerase chain reaction analysis identified the breakpoint and demonstrated the absence of the SRY gene in female members. Thus, the proband inherited this translocation from the father and grandmother. This makes the prediction of the fetal phenotype possible through assessing the grandmother. Therefore, we suggest that conventional cytogenetic and molecular cytogenetic methods, in combination with family history, provide informative results for prenatal diagnosis and prenatal genetic counseling.

Detection of genetic abnormalities in human sperm, oocytes, and preimplantation embryos using fluorescence in situ hybridization (FISH) (Fluorescence in situ hybridization(FISH) 기법을 이용한 인간 생식세포 및 착상전 배아의 유전이상 검색)

  • 방명걸
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.12-18
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    • 1998
  • Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

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Study on Evaluation of the Leak Rate for Steam Valve in Power Plant (발전용 증기밸브 누설량 평가에 관한 연구)

  • Lee, S.G.;Park, J.H.;Yoo, G.B.
    • Journal of Power System Engineering
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    • v.11 no.1
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    • pp.45-50
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    • 2007
  • Acoustic emission technology is applied to diagnosis the internal leak and operating conditions of the major valves at nuclear power plants. The purpose of this study is to verify availability of the acoustic emission as in-situ diagnosis method. In this study, acoustic emission tests are performed when the pressurized high temperature steam flowed through gate valve(1st stage reheater valve) and glove valve(main steam dump valve) on the normal size of 4 and 8". The valve internal leak diagnosis system for practical field was designed. The acoustic emission method was applied to the valves at the site, and the background noise was measured for the abnormal plant condition. To improve the reliability, a judgment of leak on the system was used various factors which are AE parameters, trend analysis, signal level analysis and RMS(root mean square) analysis of acoustic signal emitted from the valve operating condition internal leak.

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Optical In-Situ Plasma Process Monitoring Technique for Detection of Abnormal Plasma Discharge

  • Hong, Sang Jeen;Ahn, Jong Hwan;Park, Won Taek;May, Gary S.
    • Transactions on Electrical and Electronic Materials
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    • v.14 no.2
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    • pp.71-77
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    • 2013
  • Advanced semiconductor manufacturing technology requires methods to maximize tool efficiency and improve product quality by reducing process variability. Real-time plasma process monitoring and diagnosis have become crucial for fault detection and classification (FDC) and advanced process control (APC). Additional sensors may increase the accuracy of detection of process anomalies, and optical monitoring methods are non-invasive. In this paper, we propose the use of a chromatic data acquisition system for real-time in-situ plasma process monitoring called the Plasma Eyes Chromatic System (PECS). The proposed system was initially tested in a six-inch research tool, and it was then further evaluated for its potential to detect process anomalies in an eight-inch production tool for etching blanket oxide films. Chromatic representation of the PECS output shows a clear correlation with small changes in process parameters, such as RF power, pressure, and gas flow. We also present how the PECS may be adapted as an in-situ plasma arc detector. The proposed system can provide useful indications of a faulty process in a timely and non-invasive manner for successful run-to-run (R2R) control and FDC.