scholarly journals An in situ hybridization study of the effects of artificial insemination on the localization of cells expressing MHC class II mRNA in the chicken oviduct

Reproduction ◽  
2001 ◽  
pp. 581-586 ◽  
Author(s):  
WM Zheng ◽  
M Nishibori ◽  
N Isobe ◽  
Y Yoshimura
Keyword(s):  
In Situ Hybridization ◽  
Mhc Class Ii ◽  
Artificial Insemination ◽  
Laying Hens ◽  
Major Gene ◽  
Class Ii ◽  
Significant Difference ◽  
Chicken Oviduct ◽  
Fresh Semen

The aim of this study was to determine the effects of artificial insemination on the localization of antigen-presenting cells expressing MHC class II mRNA in chicken oviducts. Laying hens (35 weeks old) were inseminated with fresh semen or sham-inseminated with saline daily for 3 days. In situ hybridization was performed to detect chicken MHC class II (B-LB21 major gene) mRNA on frozen sections of oviductal infundibulum, uterovaginal junction and vagina by using digoxigenin-labelled PCR probes. Cells expressing MHC class II were observed mainly in the oviductal mucosal stroma and occasionally in the mucosal epithelium. After 24 h, the population of cells expressing MHC class II in the infundibulum was significantly higher in laying hens inseminated with fresh semen than in the control hens sham-inseminated with saline (P < 0.05). However, there was no significant difference in the population of cells expressing MHC class II in the uterovaginal junction and vagina between the artificially inseminated and control hens. These results indicate that anti-sperm immune responses, including the influx of cells expressing MHC class II and enhanced MHC class II mRNA expression, probably occur in the infundibulum after artificial insemination.

Improved Filter Method for Urine Sediment Detection of Urothelial Carcinoma by Fluorescence In Situ Hybridization

2007 ◽  
Vol 131 (10) ◽  
pp. 1574-1577 ◽  
Author(s):  
Isabelle Meiers ◽  
Harpreet Singh ◽  
Deloar Hossain ◽  
Kevin Lang ◽  
Lina Liu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Urothelial Carcinoma ◽  
Urine Cytology ◽  
Filter Method ◽  
Urine Sediment ◽  
Specific Test ◽  
Significant Difference ◽  
Time Required

AbstractContext.—Fluorescence in situ hybridization (FISH) of voided urine sediment is a sensitive and specific test for the detection of urothelial carcinoma. The time required for slide preparation using the conventional cytospin method is lengthy.Objective.—To present an alternative to the conventional cytospin method.Design.—We compared the results of an improved filter monolayer method with published results of the conventional cytospin method. A total of 624 patients with cytology and FISH analyses were followed with cystoscopy and/ or bladder biopsy. Fluorescence in situ hybridization analysis was performed on 624 cases using fluorescence-labeled probes to the pericentromeric regions of chromosomes 3, 7, and 17 and band 9p21; cytology was also performed in all cases.Results.—A total of 217 (34.7%) of 624 patients had follow-up bladder biopsies, and 170 of these (78.3%) had urothelial carcinoma. The sensitivity for cancer detection was higher for FISH than for urine cytology (92.9% [158/ 170] for FISH vs 72.9% [124/170] for urine cytology, P = &lt;5%). The specificity was equivalent for FISH and urine cytology (97.5% [443/454] for FISH vs 92.2% [419/454] for cytology). The sensitivity for FISH was better (92.9% vs 81%), and there was no significant difference in specificity (97.5% vs 96%) between the filter method and the conventional cytospin method. Unlike the conventional cytospin method, the filter method did not require multiple centrifugation and decantation steps or investment in dedicated equipment.Conclusions.—The improved filter method was faster, easier, and less expensive than published results with the conventional cytospin method with better sensitivity and equivalent specificity.

scholarly journals Modulation of Major Histocompatibility Class II Protein Expression by Varicella-Zoster Virus

2000 ◽  
Vol 74 (4) ◽  
pp. 1900-1907 ◽  
Author(s):  
Allison Abendroth ◽  
Barry Slobedman ◽  
Eunice Lee ◽  
Elizabeth Mellins ◽  
Mark Wallace ◽  
...  
Keyword(s):  
Cell Surface ◽  
Varicella Zoster Virus ◽  
Mhc Class Ii ◽  
Class Ii ◽  
Northern Blot Hybridization ◽  
Major Histocompatibility ◽  
Varicella Zoster ◽  
Ifn Γ ◽  
In Cells

ABSTRACT We sought to investigate the effects of varicella-zoster virus (VZV) infection on gamma interferon (IFN-γ)-stimulated expression of cell surface major histocompatibility complex (MHC) class II molecules on human fibroblasts. IFN-γ treatment induced cell surface MHC class II expression on 60 to 86% of uninfected cells, compared to 20 to 30% of cells which had been infected with VZV prior to the addition of IFN-γ. In contrast, cells that were treated with IFN-γ before VZV infection had profiles of MHC class II expression similar to those of uninfected cell populations. Neither IFN-γ treatment nor VZV infection affected the expression of transferrin receptor (CD71). In situ and Northern blot hybridization of MHC II (MHC class II DR-α) RNA expression in response to IFN-γ stimulation revealed that MHC class II DR-α mRNA accumulated in uninfected cells but not in cells infected with VZV. When skin biopsies of varicella lesions were analyzed by in situ hybridization, MHC class II transcripts were detected in areas around lesions but not in cells that were infected with VZV. VZV infection inhibited the expression of Stat 1α and Jak2 proteins but had little effect on Jak1. Analysis of regulatory events in the IFN-γ signaling pathway showed that VZV infection inhibited transcription of interferon regulatory factor 1 and the MHC class II transactivator. This is the first report that VZV encodes an immunomodulatory function which directly interferes with the IFN-γ signal transduction via the Jak/Stat pathway and enables the virus to inhibit IFN-γ induction of cell surface MHC class II expression. This inhibition of MHC class II expression on VZV-infected cells in vivo may transiently protect cells from CD4+ T-cell immune surveillance, facilitating local virus replication and transmission during the first few days of cutaneous lesion formation.

scholarly journals Comparative Performance of Breast Cancer Human Epidermal Growth Factor Receptor 2 Fluorescence In Situ Hybridization and Brightfield In Situ Hybridization on College of American Pathologists Proficiency Tests

2018 ◽  
Vol 142 (10) ◽  
pp. 1254-1259 ◽  
Author(s):  
Katherine B. Geiersbach ◽  
Julia A. Bridge ◽  
Michelle Dolan ◽  
Lawrence J. Jennings ◽  
Diane L. Persons ◽  
...  
Keyword(s):  
Breast Cancer ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Proficiency Testing ◽  
Copy Number ◽  
Fish Analysis ◽  
Proficiency Tests ◽  
Comparative Performance ◽  
Significant Difference

Context.— Fluorescence in situ hybridization (FISH) and brightfield in situ hybridization (ISH) are 2 clinically approved laboratory methods for detecting ERBB2 (HER2) amplification in breast cancer. Objective.— To compare the performance of FISH and brightfield ISH on proficiency testing administered by the College of American Pathologists Laboratory Accreditation Program. Design.— Retrospective review was performed on 70 tissue core samples in 7 separate proficiency testing surveys conducted between 2009 and 2013. Results.— The samples included 13 consensus-amplified tissue cores, 53 consensus-nonamplified cores, and 4 cores that did not reach consensus for FISH and/or brightfield ISH. There were 2552 individual responses for FISH and 1871 individual responses for brightfield ISH. Consensus response rates were comparable for FISH (2474 of 2524; 98.0%) and brightfield ISH (2135 of 2189; 97.5%). The FISH analysis yielded an average HER2 copy number per cell that was significantly higher (by 2.86; P = .02) compared with brightfield ISH for amplified cores. For nonamplified cores, FISH yielded slightly, but not significantly, higher (by 0.17; P = .10) HER2 copy numbers per cell. There was no significant difference in the average HER2 to control ratio for either consensus-amplified or consensus-nonamplified cores. Participants reported “unable to analyze” more frequently for brightfield ISH (244 of 2453; 9.9%) than they did for FISH (160 of 2684; 6.0%). Conclusions.— Our study indicates a high concordance rate in proficiency testing surveys, with some significant differences noted in the technical performance of these assays. In borderline cases, updated American Society of Clinical Oncology/College of American Pathologists cutoff thresholds that place greater emphasis on HER2 copy number per cell could accentuate those differences between FISH and brightfield ISH.

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 Trisomy 12 in chronic lymphocytic leukemia detected by fluorescence in situ hybridization: analysis by stage, immunophenotype, and morphology

Blood ◽  
1993 ◽  
Vol 82 (2) ◽  
pp. 571-575 ◽  
Author(s):  
TH Que ◽  
JG Marco ◽  
J Ellis ◽  
E Matutes ◽  
VB Babapulle ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chronic Lymphocytic Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Clinical Stage ◽  
Lymphocytic Leukemia ◽  
Therapeutic Trial ◽  
Number Of Patients ◽  
Significant Difference ◽  
Trisomy 12

Abstract Fluorescence in situ hybridization (FISH) with a chromosome 12 specific alpha-centromeric probe was performed on interphase cells from 183 patients with B-cell chronic lymphocytic leukemia (CLL). Twenty one cases with trisomy 12 (11.5%) were detected. The number of trisomic cells ranged from 5.5% to 76% (mean 38.5%). No correlation was found between the presence of trisomy 12 and white blood cell count, hemoglobin level, platelet count, a specific immunophenotype, clinical stage, sex, splenomegaly, or lymphadenopathy. Morphologic review of all cases with trisomy 12 showed seven (33%) with more than 10% prolymphocytes and three (14%) with CLL of mixed cell type. While trisomy 12 is the most common chromosomal abnormality in CLL, it is more frequent in morphologically atypical cases, some of which may be undergoing transformation. There was a statistically significant difference in the incidence of atypical cases between those with (47%) and without (7.6%) trisomy 12 (P < .001). It remains to be determined whether this abnormality is associated with a worse prognosis; this is currently being investigated in the context of a national therapeutic trial. The technique used is more sensitive than conventional cytogenetic analysis, which in this series failed to detect trisomy 12 in six cases. FISH allows the systematic study on a large number of patients without the need of metaphase preparations.

AMPLIFICATION AND OVEREXPRESSION OF HER2/NEU IN GASTRIC CANCER ASSESSED BY FLUORESCENCE IN SITU HYBRIDIZATION AND IMMUNOHISTOCHEMISTRY

2015 ◽  
pp. 22-31
Author(s):  
Van Huy Tran ◽  
Thi Minh Thi Ha ◽  
Viet Nho Le ◽  
Cong Thuan Dang ◽  
Trung Nghia Van ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Cancer Patients ◽  
Gene Amplification ◽  
Clinicopathologic Characteristics ◽  
Significant Difference ◽  
Her 2 ◽  
Gastric Cancer Patients

Background: HER2/neu is a predictive biomarker for treatment of gastric cancer using trastuzumab in combination with chemotherapy. This study aimed to: (1) assess the amplification and the overexpression of HER2/neu using fluorescence in situ hybridization (FISH) and immunohistochemistry in gastric cancer; (2) survey the association between HER2/neu and clinicopathologic characteristics of gastric cancer. Patients and methods: one hundred and sixty gastric cancer patients were assessed HER2/neu overexpression by IHC using Ventana anti-HER-2/neu (4B5) kit and were assessed HER2/neu gene amplification by FISH using PathVysionTM HER-2 DNA Probe kit with biopsy specimens. Results: HER2/neu protein expression rates of IHC 0, 1+, 2+ and 3+ were 70%, 10.6%, 10.6% and 8.8%, respectively. HER2/neu gene amplification was identified in gastric cancer from 21 out of 160 (13.1%) patients. The concordance between IHC and FISH was 90.0%. The HER2/neu-positive rate assessed by both techniques was 13.1%. There was a significant difference in HER2/neu-positivity between cardia gastric cancer and non-cardia gastric cancer (36.4% vs 11.4%, p = 0.040); between intestinal type and diffuse type (20.7% vs 5.9%, p = 0.010); between well, moderately differentiated and poorly differentiated gastric cancer (18.6% and 23.8% vs 5.8%, p = 0.047). Conclusion: We applied successfully FISH and IHC technique with biopsy samples in gastric cancer detecting HER2/neu positivity in order to select patients that benefit from trastuzumab in combination with chemotherapy. HER2/neu status associated with tumor location, Lauren classification and differentiated grading. Keywords: gastric cancer, fluorescence in situ hybridization (FISH), immunohistochemistry (IHC), HER-2/neu

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