Frequency of aneuploidy in pig oocytes matured in vitro and of the corresponding first polar bodies detected by fluorescent in situ hybridization

2001 ◽  
Vol 56 (5) ◽  
pp. 771-776 ◽  
Author(s):  
M. Vozdová ◽  
M. Machatková ◽  
S. Kubíčková ◽  
D. Zudová ◽  
E. Jokešová ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Polar Bodies

scholarly journals Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels

Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1502
Author(s):  
Jorge García-Hernández ◽  
Manuel Hernández ◽  
Yolanda Moreno
Keyword(s):  
In Situ Hybridization ◽  
Vibrio Parahaemolyticus ◽  
Fluorescent In Situ Hybridization ◽  
Potential Method ◽  
Viable Count ◽  
Hybridization Technique ◽  
Fish Technique ◽  
Viable Cells

Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 μg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.

scholarly journals Chromatin Quality as a Crucial Factor for the Success of Fluorescent in Situ Hybridization Analyses of Unfertilized Oocytes, Polar Bodies and Arrested Zygotes

2010 ◽  
Vol 13 (1) ◽  
pp. 1-8
Author(s):  
R Zhivkova ◽  
S Delimitreva ◽  
D Toncheva ◽  
I Vatev
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Sperm Chromatin ◽  
Fish Analysis ◽  
Premature Chromosome Condensation ◽  
Related Research ◽  
The Third ◽  
Polar Bodies ◽  
Cell Fixation

Chromatin Quality as a Crucial Factor for the Success of Fluorescent in Situ Hybridization Analyses of Unfertilized Oocytes, Polar Bodies and Arrested ZygotesMaterial that is supernumerary or unsuitable for in vitro fertilization (IVF) procedures is used for basic and for IVF-related research. Despite the disadvantages of such cells, they have contributed much to our understanding of the mechanisms and prevalence of different abnormalities.Fifty-four human unfertilized oocytes, 34 arrested bipronuclear zygotes and 15 polar bodies were fixed for analysis on the third day after in vitro insemination and were subjected to fluorescent in situ hybridization (FISH) with probes for chromosomes 18, 21, X and Y (centromere for 18, X, Y and locus-specific for 21). The aim of the study was the comparison of FISH efficiency in differently condensed chromatin.The success of FISH analysis was over 60% of analyzed cells and it was dependent on the chromatin changes (condensation and/or fragmentation) during the culture period before cell fixation. Chromatin ageing was the crucial factor for the reduced success of FISH in both oocyte chromosomes (60.0%) and pronuclei (61.76%). The chromatin of second polar bodies (PBII), and premature chromosome condensation (PCC) of the sperm chromatin in oocytes was more suitable for FISH analysis (FISH success 75.0% in PBII and 64.29% in PCC) with both centromere and locus-specific probes.These results revealed the significance of early signs of in vitro cell ageing for the success of FISH analysis and for the interpretation of results in case of analysis of unfertilized human ova, polar bodies and arrested zygotes.

Detection of aneuploidy in human oocytes and corresponding first polar bodies by fluorescent in situ hybridization

1996 ◽  
Vol 13 (1) ◽  
pp. 73-78 ◽  
Author(s):  
Andrei Dyban ◽  
Michael Freidine ◽  
Elena Severova ◽  
Jeanine Cieslak ◽  
Victor Ivakhnenko ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Human Oocytes ◽  
Polar Bodies

scholarly journals STUDY OF CYTOGENETIC ABNORMALITIES IN G-CSF STIMULATED PERIPHERAL BLOOD CELLS AND NON-STIMULATED BONE MARROW CELLS OF PATIENTS WITH MYELOFIBROSIS

2016 ◽  
Vol 38 (1) ◽  
pp. 40-44 ◽  
Author(s):  
R Y Lozynskyy ◽  
M R Lozynska ◽  
Y V Hontar ◽  
N L Huleyuk ◽  
Z V Maslyak ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Situ Hybridization ◽  
Peripheral Blood ◽  
Fluorescent In Situ Hybridization ◽  
Chromosomal Abnormalities ◽  
Cytogenetic Study ◽  
Hybridization Method ◽  
Cytogenetic Abnormalities

The aim of the study was to improve cytogenetic diagnostics and monitoring of myelofibrosis and to reveal the spectrum of cytogenetic abnormalities in patients from Ukraine. Materials and Methods: A total of 42 patients (23 females and 19 males) with myelofibrosis was studied using different cytogenetic methods. Granulocyte colony-stimulating factor (G-CSF) was added by the new method during cultivation of peripheral blood (PB) cells from 31 patients for specific stimulation of mitotic divisions. Two patients underwent examination by fluorescent in situ hybridization method. Results: In cell cultures of PB stimulated in vitro with G-CSF and in non-stimulated bone marrow chromosome abnormalities were found in 19 (45.2%) of all the patients. The spectrum of cytogenetic abnormalities of bone marrow and PB was the same in all of the patients. Aspiration of bone marrow was unsuccessful due to significant fibrosis in 10 (29.4%) of 34 patients. The study by fluorescent in situ hybridization method confirmed cytogenetic abnormalities revealed by G-method and discovered additional possibly normal subclone. Conclusions: Cytogenetic study of PB using in vitro G-CSF as a specific stimulant of mitosis instead of phytohemagglutinin revealed significant variety of chromosomal abnormalities in Ukrainian patients with myelofibrosis. This method could be a less invasive alternative to cytogenetic examination of bone marrow in the subgroup of patients with considerable fibrosis and consecutive changes. The usage of fluorescent in situ hybridization method supplemented karyotyping by G-banding method.

scholarly journals Visualization of Alternative Epstein-Barr Virus Expression Programs by Fluorescent In Situ Hybridization at the Cell Level

1999 ◽  
Vol 73 (6) ◽  
pp. 5064-5069 ◽  
Author(s):  
Anna Szeles ◽  
Kerstin I. Falk ◽  
Stephan Imreh ◽  
George Klein
Keyword(s):  
In Situ Hybridization ◽  
Cell Lines ◽  
Fluorescent In Situ Hybridization ◽  
Epstein Barr Virus ◽  
Type I ◽  
Type Iii ◽  
Barr Virus ◽  
Epstein Barr

ABSTRACT Epstein-Barr virus (EBV) transforms human B lymphocytes into immortalized lymphoblastoid cell lines (LCLs). They regularly express six virally encoded nuclear proteins (EBNA1 to EBNA6) and three membrane proteins (LMP1, LMP2A, and LMP2B). In contrast, EBV-carrying Burkitt lymphoma (BL) cells in vivo and derived type I cell lines that maintain the BL phenotype express only EBNA1. During prolonged in vitro culturing, most EBV-carrying BL lines drift toward a more immunoblastic (type II or III) phenotype. Their viral antigen expression is upregulated in parallel. We have used fluorescent in situ hybridization to visualize viral transcripts in type I and III BL lines and LCLs. In type I cells, EBNA1 is encoded by a monocistronic message that originates from the Qp promoter. In type III cells, the EBNA1 transcript is spliced from a giant polycistronic message that originates from one of several alternative Wp or Cp promoters and encodes all six EBNAs. We have obtained a “track” signal with aBamHI W DNA probe that could hybridize with the polycistronic but not with the monocistronic message in two type III BL lines (Namalwa-Cl8 and MUTU III) and three LCLs (LCL IB4-D, LCL-970402, and IARC-171). A BamHI K probe that can hybridize to both the monocistronic and the polycistronic message visualized the same pattern in the type III BLs and the LCLs as the BamHI W probe. A positive signal was obtained with the BamHI K but not the BamHI W probe in the type I BL lines MUTU I and Rael. The RNA track method can thus distinguish between cells that use a type III and those that use a type I program. The former cells hybridize with both the W and the K probes, but the latter cells hybridize with only the K probe. Our findings may open the way for studies of the important but still unanswered question of whether cells with type I latency arise from immunoblasts with a full type III program or are generated by a separate pathway during primary infection.

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