scholarly journals Transcriptome Analysis and In Situ Hybridization for FcaGHV1 in Feline Lymphoma

Viruses ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 464 ◽  
Author(s):  
Mahdis Aghazadeh ◽  
Mang Shi ◽  
Patricia Pesavento ◽  
Amy Durham ◽  
Tamsen Polley ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Cellular Transformation ◽  
Clinical Samples ◽  
Domestic Cats ◽  
Reverse Transcription Pcr ◽  
Low Copy Number ◽  
Intestinal Lymphomas ◽  
Lytic Genes ◽  
Common Infection

Lymphoma is one of the most common malignancies in domestic cats. The lymphomagenicpotential of Felis catus gammaherpesvirus 1 (FcaGHV1), a common infection in domestic cats,is unknown. In other species, including humans, cellular transformation by gammaherpesvirusesis typically mediated by viral genes expressed during latency. We analysed tumour RNA, fromdiffuse large B-cell lymphomas (DLBCL) appearing in cats coinfected with FcaGHV1 and felineimmunodeficiency virus (FIV) (n = 10). Analysis was done by high throughput transcriptomesequencing and reverse transcription PCR. A limited repertoire of FcaGHV transcripts was identifiedin five tumors, including homologs of oncogenic latency-associated transcripts, latency-associatednuclear antigen (LANA, ORF73) and vFLIP (F7), lytic genes (ORF50, ORF6, ORF59, F10), and an ORFunique to FcaGHV1, F20. In situ hybridization of FIV-associated DLBCLs (n = 9), post-transplantlymphomas (n = 6) and high-grade B and T-cell intestinal lymphomas (n = 8) identified a single casein which FcaGHV1 nucleic acid was detectable. These results demonstrate that FcaGHV1 transcriptscan be detected in some FIV-associated lymphomas, but with a low copy number, precludingassessment of a potential role for FcaGHV1 in lymphomagenesis. Future investigation of the FcaGHV1transcriptome in clinical samples might employ viral enrichment and greater sequencing depth toenhance the retrieval of viral reads. Our results suggest prioritization of a subset of intestinal T-celltumors and large granular lymphocyte lymphoma, for study.

Comet and cup genes in Drosophila spermatogenesis: the first demonstration of post-meiotic transcription

2008 ◽  
Vol 36 (3) ◽  
pp. 540-542 ◽  
Author(s):  
Carine Barreau ◽  
Elizabeth Benson ◽  
Helen White-Cooper
Keyword(s):  
In Situ Hybridization ◽  
Expression Pattern ◽  
Reverse Transcription ◽  
Protein Product ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Single Cyst ◽  
Reverse Transcription Pcr

Post-meiotic transcription is widespread in mammalian spermatogenesis, but is generally believed to be absent from Drosophila spermatogenesis. Genes required during meiosis, in early spermatids or later in spermiogenesis are typically transcribed in primary spermatocytes in Drosophila. Their mRNAs are then stored in the cytoplasm until the protein product is needed. Recently, using in situ hybridization, we identified 17 Drosophila genes, collectively named ‘comets’ and ‘cups’, whose mRNAs are most abundant in, and localize to the distal ends of, elongating spermatids. Using a single-cyst quantitative RT–PCR (reverse transcription–PCR) assay, we confirmed this unusual expression pattern and conclusively demonstrate the existence of post-meiotic transcription in Drosophila spermatids. We found that transcription of comets and cups occurs just before protamines can be detected in spermatid nuclei.

scholarly journals Combining TrkA Immunohistochemical-Score with Clinicopathologic Parameters in Neuroblastoma: An Influential Prognostic Nomogram

2021 ◽  
Author(s):  
Juanqing Yue ◽  
Lei Cai ◽  
Ruifen Wang ◽  
Meng Qiao ◽  
Kezhou Wang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Prognostic Factor ◽  
Fluorescence In Situ Hybridization ◽  
Gene Fusion ◽  
Independent Prognostic Factor ◽  
Clinical Samples ◽  
Qrt Pcr ◽  
Clinicopathologic Parameters ◽  
Low Expression

Abstract BackgroundNeuroblastoma (NB) is one of the most common solid tumors in children with varied clinical outcomes. Although there are some several risk stratification systems currently, their clinical applications are limited due to the testing conditions of different laboratory and the heavy financial burden on patients. TrkA is coded by NTRK1 , belonging to tropomyosin receptor kinase family. We have observed that TrkA was differentially expressed in paraffin tissue sections of NB. The aim of this study was to determine the immunohistochemical-score of TrkA as an independent prognostic factor for NB and establish a useful prognostic model for postoperative patients.MethodsWe systematically summarized the relationship between immunochemistry (IHC) score of TrkA and clinicopathological parameters in 86 NB cases. Fluorescence in situ hybridization (FISH) and qRT-PCR were used to detect NTRK1 gene fusion. Furthermore, GSE96631, GSE16476, GSE49710 and GSE73537 datasets, originated from Gene Expression Omnibus (GEO), were analyzed to figure out the NTRK1 related molecular characteristics by bioinformatics methods. And combined TrkA immunohistochemical-score with clinicopathologic parameters to construct a prognostic nomogram of overall survival (OS) for NB.ResultIn clinical samples and GEO database analyses, patients in the NTRK1 / TrkA low expression group showed significantly poorer outcome than patients in high group. Multivariate cox regression analysis demonstrated NTRK1 / TrkA as an independent prognostic factor for NB survival. Neither Fluorescence in situ hybridization nor qRT-PCR detected evidence for NTRK1 gene fusion in clinical samples, indicating that differential expression in NTRK1 / TrkA are caused by epigenetic changes. Bioinformatics analyses revealed that MYC target related pathway may play a critical role in low expression of TrkA, leading to unfavorable prognoses of NB.ConclusionThe results of this study suggest that the IHC score of TrkA may be used as an independent predictor of postoperative OS of patients with NB. By combining the IHC score of TrkA and clinicopathological features, the proposed nomogram provides a feasible predictive tool for postoperative patients with NB. Simultaneously, this study also reveals that Trk inhibitors are not supposed to be taken in NB patients.

scholarly journals DETECTION AND QUANTITATION OF RED COMPLEX BACTERIA IN SUBGINGIVAL PLAQUE BY USING FLUORESCENT IN SITU HYBRIDIZATION (FISH)

2017 ◽  
Vol 5 (11) ◽  
pp. 279-289
Author(s):  
Kishore G. Bhat ◽  
Aradhana Chhatre ◽  
Vijay M. Kumbar ◽  
Manohar S. Kugaji ◽  
Sanjeevani Patil
Keyword(s):  
In Situ Hybridization ◽  
Diagnostic Tool ◽  
Fluorescent In Situ Hybridization ◽  
High Sensitivity ◽  
Clinical Samples ◽  
Subgingival Plaque ◽  
Periodontal Pathogens ◽  
Significant Difference ◽  
Strong Linear Relationship

Motivation/Background: Red complex bacteria are proven periodontal pathogens. In dentistry, there is a need to identify and quantitate the organisms from the diseased sites quickly and reliably. Since culture requires several days, molecular methods are being used frequently to detect these bacteria.  Among them, Fluorescent in situ hybridization (FISH) is rapid, sensitive and quantitative. An attempt is made here to evaluate the applicability of this technique as a diagnostic tool in periodontology. Method: Subgingival plaque was collected from participants, fixed with paraformaldehyde and subjected to FISH. Fluorescently labeled oligonucleotide probes were used for hybridization. After the procedure, the fluorescently stained bacteria were identified and counted from the smear and quantitated using a simple grading. Results: There was a significant difference in the prevalence and numbers of red complex bacteria in healthy and diseased subjects. A strong linear relationship existed between P. gingivalis, T. forsythia and T. denticola. Conclusions: The procedure used in the study is simple, rapid and can be easily adaptable. It also has a high sensitivity and has the ability to detect a single bacterial cell. The method can be directly applied to the clinical samples and can be used as a rapid diagnostic tool in periodontics.

scholarly journals Prolactin Receptor Messenger Ribonucleic Acid in Normal and Neoplastic Human Pituitary Tissues1

1997 ◽  
Vol 82 (3) ◽  
pp. 963-968
Author(s):  
Long Jin ◽  
Xiang Qian ◽  
Elzbieta Kulig ◽  
Bernard W. Scheithauer ◽  
Rocio Calle-Rodrigue ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Reverse Transcription ◽  
Ribonucleic Acid ◽  
Prolactin Receptor ◽  
Pituitary Surgery ◽  
Specific Cell ◽  
Human Pituitary ◽  
Messenger Ribonucleic Acid ◽  
Reverse Transcription Pcr

Abstract We examined the specific cell types in normal human pituitaries that expressed PRL receptor (PRL-R) messenger ribonucleic acid (mRNA) by combined in situ hybridization and immunohistochemistry. The distribution of PRL-R mRNA in 28 pituitary adenomas was examined by in situ hybridization and reverse transcription-PCR in 12 cases of adenomas. In another set of experiments, 34 PRL adenomas from men, women, and bromocriptine-treated patients were analyzed for PRL-R by in situ hybridization. In the normal pituitary, PRL- and LH-producing cells had significantly more mean grain counts per cell and higher percentages of cells positive for PRL-R than GH and TSH cells. PRL-R mRNA was present in all groups of adenomas by in situ hybridization and reverse transcription-PCR. PRL adenomas had a significantly higher density of labeling compared to other adenoma types. Although there was no difference in the levels of PRL-R mRNA in PRL adenomas from men and premenopausal and postmenopausal women, patients treated with bromocriptine before pituitary surgery had significantly lower levels of PRL-R compared to all other groups. These results indicate that in the normal pituitary, PRL and LH cells have the highest level of PRL-R mRNA, whereas PRL adenomas have significantly higher levels of PRL-R mRNA than other types of adenomas, and bromocriptine treatment decreases the levels of PRL-R mRNA in PRL adenomas.

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