scholarly journals Chromogenic in Situ Hybridization for Human Cytomegalovirus-DNA Detection in Tissue Subsets with Prostatic Adenocarcinoma and Benign Hyperplasia

2021 ◽  
pp. 2894-2905
Author(s):  
Tayseer Anmar Hassan ◽  
Jenan M. Jawad AL-Saffar ◽  
Saad Hasan Mohammed Ali
Keyword(s):  
In Situ Hybridization ◽  
Benign Prostatic Hyperplasia ◽  
Human Cytomegalovirus ◽  
Dna Detection ◽  
Prostatic Hyperplasia ◽  
Prostatic Adenocarcinoma ◽  
Wide Range ◽  
Chromogenic In Situ Hybridization ◽  
Benign Hyperplasia

     Human cytomegalovirus (HCMV) infects a wide range of human cells, resulting in both benign and malignant tumors. In the last few decades, proteins and/or nucleic acids of the virus were found to be often highly expressed in in patients with basal cell hyperplasia and prostatic neoplasia. This research aimed to unravel the rate of HCMV infections among prostatic tissue subsets from Iraqi patients with adenocarcinoma and benign hyperplasia. One hundred, formalin-fixed and paraffin embedded prostatic tissues were obtained from 40 tissue samples collected from different grades of prostate carcinoma; 40 from benign prostatic hyperplasia and 20 from apparently healthy prostatic tissues. These tissue specimens were collected from the archives of different public and private histopathological laboratories in Baghdad. Detection of HCMV-DNA was achieved by a highly sensitive version of chromogenic in situ hybridization technique. The signals of chromogenic in situ hybridization reactions for HCMV-DNA detection in prostatic adenocarcinoma tissues were found in 65% (26 out of 40) of the tissues, whereas in BPH (Benign Prostatic Hyperplasia), HCMV-DNA was detected in 57.5% (23 out of 40) of the tissues, and in the healthy control group in 25% (5 out of 20) of the tissues. The highest percentage of positive- HCMV- DNA-CISH reactions (57.5%) was found in prostatic adenocarcinomatous tissues that showed poor differentiation. Our results could show that HCMV might contribute to the development of the studied subsets of prostatic adenocarcinoma and benign prostatic hyperplasia.

scholarly journals Molecular Detection of Human Herpes Virus-8 in Prostatic Adenocarcinoma and Benign Prostatic Hyperplasia Tissues by DNA -In Situ Hybridization

2021 ◽  
pp. 96-107
Author(s):  
Zainab Mahdi Saleh ◽  
Jinan Al-Saffar ◽  
Saad Hassan
Keyword(s):  
In Situ Hybridization ◽  
Benign Prostatic Hyperplasia ◽  
Prostatic Hyperplasia ◽  
Prostatic Adenocarcinoma ◽  
Human Herpes ◽  
Human Herpes Virus 8 ◽  
Human Herpes Virus ◽  
Group Detection ◽  
Herpès Virus

Human Herpes Virus-8 (HHV-8) is a sexually transmitted viral infection that can infect the prostate epithelium in immunocompromised adults. Recently, HHV-8 was related to the development and progression of several human malignancies like prostatic adenocarcinoma. This retrospective research was designed to analyze the distribution and possible impact of HHV-8 infection on prostatic adenocarcinogenesis. A total number of one hundred formalin-fixed prostatic tissues were enrolled in this research; forty Prostate Adenocarcinoma (PAC) biopsies, forty biopsies from Benign Prostatic Hyperplasia (BPH), and twenty Apparently Normal Prostatic Tissues (ANPT) as a control group. Detection of HHV -8 DNA was achieved by a highly-sensitive variant of Chromogenic In Situ Hybridization (CISH) technique. In the mean age of PAC patients was 64.08±7.54years. Detection of CISH reactions for HHV 8- DNA was observed in tissues of 70% (28 out of 40) of PAC patients and 30% (12 out of 40) of BPH tissues, whereas no positive reactions were detected in the ANPT group. Detection of CISH reactions for HHV 8- DNA was observed in 55.6%  of tissues of PAC patients  with well grade histopathological examination,  87.5% of moderate grade, and 69.6% of poorly differentiated grade. It can be concluded that HHV-8 infection might contribute in prostate oncogenesis, together with other essential oncogenic viruses.

scholarly journals Zone-Dependent Expression of Estrogen Receptors α and β in Human Benign Prostatic Hyperplasia

2003 ◽  
Vol 88 (3) ◽  
pp. 1333-1340 ◽  
Author(s):  
Toshifumi Tsurusaki ◽  
Daiyu Aoki ◽  
Hiroshi Kanetake ◽  
Satoshi Inoue ◽  
Masami Muramatsu ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
In Situ Hybridization ◽  
Benign Prostatic Hyperplasia ◽  
Estrogen Receptors ◽  
Stromal Cells ◽  
Crucial Role ◽  
Peripheral Zone ◽  
Prostatic Hyperplasia ◽  
Cellular Distribution

Estrogen, which acts through estrogen receptors (ERs) α and β, has been implicated in the pathogenesis of benign and malignant human prostatic tumors, i.e. benign prostatic hyperplasia and prostate cancer, thought to originate from different zones of the prostate [the transition zone (TZ) and peripheral zone (PZ), respectively]. Here, we examined the cellular distribution of ERα and ERβ in human normal and hyperplastic prostate tissues, using in situ hybridization and immunohistochemistry. ERα expression was restricted to stromal cells of PZ. In contrast, ERβ was expressed in the stromal cells of PZ as well as TZ. ERβ-positive epithelial cells were evenly distributed in PZ and TZ of the prostate. Our results suggest that estrogen may play a crucial role in the pathogenesis of benign prostatic hyperplasia through ERβ.

scholarly journals Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

BMC Cancer ◽  
2009 ◽  
Vol 9 (1) ◽  
Author(s):  
Fabíola E Rosa ◽  
Sara M Silveira ◽  
Cássia GT Silveira ◽  
Nádia A Bérgamo ◽  
Francisco A Moraes Neto ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Breast Carcinoma ◽  
Real Time ◽  
Rt Pcr ◽  
Her 2 ◽  
Chromogenic In Situ Hybridization

scholarly journals Evaluating HER2 Gene Amplification Using Chromogenic In Situ Hybridization (CISH) Method In Comparison To Immunohistochemistry Method in Breast Carcinoma

2018 ◽  
Vol 6 (11) ◽  
pp. 1977-1981 ◽  
Author(s):  
Hadi Atabati ◽  
Amir Raoofi ◽  
Abdollah Amini ◽  
Reza Masteri Farahani
Keyword(s):  
Breast Cancer ◽  
In Situ Hybridization ◽  
Gene Amplification ◽  
Method Comparison ◽  
Her2 Expression ◽  
Her2 Gene ◽  
Cross Sectional ◽  
Exact Test ◽  
Chromogenic In Situ Hybridization

BACKGROUND: In patients with breast cancer, HER2 gene expression is of a great importance in reacting to Herceptin treatment. To evaluate this event, immunohistochemistry (IHC) has been done routinely on the basis of scoring it and so the patients were divided into 4 groups. Lately, as there have been disagreements about how to treat score 2 patients, chromogenic in situ hybridization (CISH) and florescence in situ hybridization (FISH) are introduced. Since CISH method is more convenient than FISH for gene amplification study, FISH has been substituted by CISH. AIM: The current study is conducted in order to investigate whether using CISH is a better method comparison to IHC method for determines HER2 expression in patients with breast cancer in. METHODS: In this cross-sectional descriptive analytical study, information of 44 female patients with invasive ductal breast cancer were gathered from Imam Reza and Omid Hospital in Mashhad. IHC staining was done for all patients in order to determine the level of HER2 expression, and after scoring them into 4 groups of 0, +1, +2 and +3, CISH staining was carried out for all 4 groups. At the end, results from both methods were statistically evaluated using SPSS software V.22.0. RESULTS: The average age of patients was 50.2 with the standard deviation of 10.96. Using IHC method was observed that 2.6% (1 patient), 26.3% (10 patients), 65.8% (25 patients) and 5.3% (2 patients) percentage of patients had scores of 0, +1, +2 and +3. On the other hand, CISH method showed 36 patients (90%) with no amplifications and 4 (10%) with sever amplifications. In a comparative study using Fisher's exact test (p = 0.000), we found a significant relation between IHC method and CISH method indicating that all patients showing severe amplifications in CISH method, owned scores of +2 and +3 in IHC method. CONCLUSION: According to the present study and comparing the results with similar previous studies, it can be concluded that CISH method works highly effective in determining HER2 expression level in patients with breast cancer. This method is also able to determine the status of patients with score +2 in IHC for their treatment with herceptin

scholarly journals Influence of prolonged formalin fixation of tissue samples on the sensitivity of chromogenic in situ hybridization

2011 ◽  
Vol 23 (6) ◽  
pp. 1212-1216 ◽  
Author(s):  
Meike M. Mostegl ◽  
Barbara Richter ◽  
Nora Dinhopl ◽  
Herbert Weissenböck
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Signal Intensity ◽  
Formalin Fixation ◽  
Proteinase K ◽  
Cross Linking ◽  
Fixation Time ◽  
Tissue Samples ◽  
Chromogenic In Situ Hybridization

Chromogenic in situ hybridization (ISH) is a commonly used tool in diagnostic pathology to detect pathogens in formalin-fixed, paraffin-embedded (FFPE) tissue sections. Prolonged formalin fixation time was identified to be a limiting factor for the successful detection of nucleic acid from different pathogens, most probably due to the cross-linking activity of formalin between RNA, DNA, and proteins. Therefore, in the current study, the influence of formalin fixation time on ISH signal intensity of 2 viral ( Porcine circovirus-2 [PCV-2] and Porcine respiratory and reproductive virus [PRRSV]) and 2 protozoal agents ( Cryptosporidium serpentis and Tritrichomonas sp.) was evaluated. Tissue samples were fixed in 7% neutral buffered formaldehyde solution, and at defined intervals, pieces were embedded in paraffin wax and subjected to pathogen-specific ISH. For all 4 pathogens, the signal intensity remained comparable with the starting ISH signal for different periods of fixation (PCV-2: 6 weeks, PRRSV: 23 weeks, C. serpentis: 55 weeks, Tritrichomonas sp.: 53 weeks). Thereafter, the signal started to decline until loss of nucleic acid detection. The influence of increased proteinase K concentrations for inverting the formalin-induced cross-linking activity was examined compared with the standard protocol. With all 4 infectious agents, a 4-fold proteinase K concentration restored the ISH signals to a level comparable with 1 day of fixation. In conclusion, the influence of prolonged formalin fixation on the intensity of detected ISH signal highly depends on the analyzed infectious agent and the pretreatment protocol.

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