Mapping of the porcine ? interferon (IFNA) gene to Chromosome 1 by fluorescence in situ hybridization

1993 ◽  
Vol 4 (1) ◽  
pp. 62-63 ◽  
Author(s):  
Ulla M. Sarmiento ◽  
Juan I. Sarmiento ◽  
Joan K. Lunney ◽  
Shanta S. Rishi
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome 1 ◽  
Ifna Gene

scholarly journals Fluorescence in situ hybridization in diluted and flow cytometrically sorted boar spermatozoa using specific DNA direct probes labelled by nick translation

Reproduction ◽  
2003 ◽  
pp. 317-325 ◽  
Author(s):  
I Parrilla ◽  
JM Vazquez ◽  
M Oliver-Bonet ◽  
J Navarro ◽  
J Yelamos ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Internal Control ◽  
Dna Probes ◽  
Chromosome 1 ◽  
Nick Translation ◽  
Fluorescent Signal ◽  
Sperm Separation

Successful evaluation of X- and Y-chromosome-bearing sperm separation technology using flow cytometry-cell sorter is of great importance. Fluorescence in situ hybridization (FISH), which allows for the detection of specific nucleic acid sequences on morphologically preserved spermatozoa, is an ideal method for quantitatively and qualitatively assessing the purity of sorted sperm samples. In this study specific pig DNA direct probes for small regions of chromosomes 1 and Y were used. Chromosome 1 was labelled in green and used as internal control to detect a lack of hybridization, whereas chromosome Y was labelled in red. Nick translation was used as the labelling method for the preparation of these probes. Spermatozoa, unsorted and sorted for high and low Y-chromosome purity from ejaculates of five boars, were fixed on slides and two-colour direct FISH was performed for chromosomes 1 and Y. About 500 non-sorted and 200 sorted spermatozoa per sample were scored. The proportion of Y-chromosome-bearing spermatozoa was determined by the presence of a red fluorescent signal on the sperm head and the proportion of X-chromosome-bearing spermatozoa was determined by subtraction. The efficiency of the hybridization procedure was established as near 98% on sorted and unsorted samples. The results of this study confirm that direct FISH using specific pig DNA probes labelled by nick translation provides a useful tool for laboratory validation of sperm separation by flow sorting technology. Moreover, the ease of nick translation and the quality of the fluorescent signal obtained using this method makes this procedure the most appropriate method for labelling pig DNA probes to be used for direct FISH on pig spermatozoa.

An evaluation of a new series of fluorescent dUTPs for fluorescence in situ hybridization.

1996 ◽  
Vol 44 (5) ◽  
pp. 525-529 ◽  
Author(s):  
J Wiegant ◽  
N Verwoerd ◽  
S Mascheretti ◽  
M Bolk ◽  
H J Tanke ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Human Lymphocytes ◽  
Single Copy ◽  
Cyanine Dye ◽  
Chromosome 1 ◽  
Nick Translation ◽  
Qualitative Comparison ◽  
Copy Dna

Synthesis of fluorochrome-modified deoxyribonucleotides has been carried out mostly by linking the fluorochrome molecule to the C-5 position of dUTP via an allylamine spacer, similar to the modification of allylamine-dUTP with the haptens biotin and digoxigenin. Recently, a new series of fluorescent nucleotides has been prepared by using an alkynyl bridge between the uracil moiety and the fluorochrome. Here we report the qualitative and quantitative analysis of fluorescence in situ hybridization results obtained on interphase cells and chromosomes with a variety of highly repetitive and single-copy DNA probes that were modified by nick translation with such alkynyl dUTPs. A qualitative comparison was made of the alkynyl dUTPs conjugated to the fluorochromes fluorescein, the cyanine dye Cy3, tetramethylrhodamine, Lissamine and Texas Red. With the exception of tetramethylrhodamine, all fluorochromes performed satisfactorily. The cyanine dye Cy3 provided the highest sensitivity, i.e., cosmid and YAC probes could easily be visualized by conventional fluorescence microscopy. In a quantitative assay, different nick translation conditions were tested using a human chromosome 1 satellite III probe (pUC1.77) and alkynyl dUTPs labeled with fluorescein and Cy3. Using these two nucleotides, FISH signal intensities on interphase nuclei from human lymphocytes were quantitated by digital imaging microscopy. The strongest signals were obtained when during nick translation the ratio between dTTP and fluorescein-dUTP or Cy3-dUTP was 1:5.

Detection of chromosomal anomalies in uterine endometrial carcinoma using fluorescence in situ hybridization (UteroFISH)

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 5533-5533
Author(s):  
J. Qian ◽  
D. Weber ◽  
R. Cochran ◽  
D. Hossain ◽  
D. G. Bostwick
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Endometrial Carcinoma ◽  
Endometrial Adenocarcinoma ◽  
Atypical Hyperplasia ◽  
Chromosome 1 ◽  
Chromosomal Anomalies ◽  
Tissue Samples ◽  
Gynecological Malignancy

5533 Background: Endometrial cancer is the most common pelvic gynecological malignancy. The diagnosis of well-differentiated endometrial adenocarcinoma, atypical hyperplasia, and marked hyperplasia is often challenging. We sought to investigate the utility of chromosomal anomalies for the detection of uterine endometrial carcinoma using multitarget fluorescence in situ hybridization (FISH). Methods: Samples were collected by endometrial brush and processed by liquid-based thin-layer cytological preparation protocol. For study, we collected cytology slides from consecutive cases to include 50 benign, 50 hyperplasia without atypia, 50 atypical hyperplasia, and 50 endometrial cancers. Each was hybridized using fluorescence labeled DNA probes to chromosomes 1, 8, and 10 (UteroFISH). The FISH signals were enumerated in 100 cells per case, and the chromosomal anomalies were correlated with pathologic findings, including histologic diagnoses on endometrial tissue samples. Results: Numeric chromosomal anomalies were found in 0% (0/50) of benign, 20% (10/50) of hyperplasia, 76% of atypical hyperplasia (38/50), and 86% (43/50) of carcinoma specimens. The mean percentage of cells with chromosomal changes was 54% in cancer specimens, significantly higher than that in hyperplasia without atypia (13%, p< 0.0001) and atypical hyperplasia (34%, p< 0.0001). The most frequent chromosomal anomaly was gain of chromosome 1. FISH anomalies had an overall sensitivity of 81% and specificity of 90% for the detection of atypical hyperplasia and/or endometrial carcinoma. There was no association with grade of endometrial carcinoma. Conclusions: Multi-target UteroFISH appeared to be useful for the differential diagnosis of reactive hyperplasia, atypical hyperplasia, and endometrial adenocarcinoma, with a high level of sensitivity and specificity. Endometrial hyperplasia with FISH-detected chromosomal anomalies may require close clinical follow-up. No significant financial relationships to disclose.

Physical organization of the major duplication on Brassica oleracea chromosome O6 revealed through fluorescence in situ hybridization with Arabidopsis and Brassica BAC probes

Genome ◽  
2005 ◽  
Vol 48 (6) ◽  
pp. 1093-1103 ◽  
Author(s):  
E C Howell ◽  
S J Armstrong ◽  
G C Barker ◽  
G H Jones ◽  
G J King ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Brassica Oleracea ◽  
Artificial Chromosome ◽  
Genetic Distances ◽  
Chromosome 1 ◽  
Cytogenetic Map ◽  
Inverted Duplication ◽  
Close Relationship

The close relationship between Brassica oleracea and Arabidopsis thaliana has been used to explore the genetic and physical collinearity of the two species, focusing on an inverted segmental chromosome duplication within linkage group O6 of B. oleracea. Genetic evidence suggests that these segments share a common origin with a region of Arabidopsis chromosome 1. Brassica oleracea and Arabidopsis bacterial artificial chromosome probes have been used for fluorescence in situ hybridization analysis of B. oleracea pachytene chromosomes to further characterize the inverted duplication. This has been highly effective in increasing the local resolution of the cytogenetic map. We have shown that the physical order of corresponding genetic markers is highly conserved between the duplicated regions in B. oleracea and the physical lengths of the regions at pachytene are similar, while the genetic distances are considerably different. The physical marker order is also well conserved between Arabidopsis and B. oleracea, with only one short inversion identified. Furthermore, the relative physical distances between the markers in one segment of B. oleracea and Arabidopsis have stayed approximately the same. The efficacy of using fluorescence in situ hybridization, together with other forms of physical and genetic mapping, for elucidating such issues relating to synteny is discussed.Key words: collinearity, cytogenetic map, pachytene chromosomes, Brassica, Arabidopsis.

scholarly journals Treatment of Leptomeningeal Metastases Evaluated by Interphase Cytogenetics

2000 ◽  
Vol 18 (10) ◽  
pp. 2053-2058 ◽  
Author(s):  
R.J. van Oostenbrugge ◽  
A.H. N. Hopman ◽  
J.W. Arends ◽  
F.C. S. Ramaekers ◽  
A. Twijnstra
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Leptomeningeal Metastases ◽  
Response To Therapy ◽  
Chromosome 1 ◽  
Interphase Cytogenetics ◽  
Cytologic Examination ◽  
Neurologic Status ◽  
Two Samples

PURPOSE: Although cytologic examination of CSF is the primary method for the evaluation of response to therapy for leptomeningeal metastases (LMMs), the procedure’s sensitivity decreases throughout the course of protracted therapy. We studied whether this response could be monitored more accurately through the detection of numerical chromosomal aberrations by interphase cytogenetics, using fluorescence in situ hybridization (FISH). PATIENTS AND METHODS: Seven patients treated for LMMs and with a known numerical aberration for chromosome 1 in their pretreatment CSF were included in this study. Up to 16 consecutive CSF samples were analyzed by means of the fluorescence in situ hybridization (FISH) technique for cells with aberrant chromosome 1 content. The results of routine cytology and FISH analyses were compared and were correlated with each patient’s neurologic status. RESULTS: Routine cytology detected malignancies in only 24 of the 76 samples, all of which were classified as chromosomally abnormal by FISH (except for two samples that could not be evaluated). Moreover, FISH demonstrated aneusomic cells in 32 additional samples, which could therefore be classified as malignant. The FISH results correlated better with patient neurologic status in that more malignant cells were detected in the CSF of neurologically deteriorating patients. CONCLUSION: Using FISH in addition to performing routine cytologic examination of CSF led to a more accurate evaluation of response to treatment in patients treated for LMMs.

Absence of Cytogenetic Abnormalities of PRV-1, TPO and C-MPL Genes Detected by Fluorescence In Situ Hybridization in Essential Thrombocythemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4746-4746
Author(s):  
Eulalia Puigdecanet ◽  
Blanca Espinet ◽  
Olaya Villa ◽  
Lurdes Zamora ◽  
Carles Besses ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Essential Thrombocythemia ◽  
Leukemia Virus ◽  
Normal Karyotype ◽  
Chromosome 1 ◽  
Interphase Nuclei ◽  
Cytogenetic Abnormalities

Abstract Introduction. Essential thrombocythemia (ET) is a chronic myeloproliferative disorder (CMPD) with heterogeneous features and no specific diagnostic markers. Consequently, its diagnosis is based on exclusion of other CMPD and secondary thrombocytosis. The role of PRV-1 (Polycythemia rubra vera-1), TPO (Thrombopoietin) and c-Mpl (Myeloproliferative leukemia virus oncogene) in ET pathogenesis has been studied in order to find new molecular targets which would help in ET diagnosis. PRV-1 gene is overexpressed in granulocytes from polycythemia vera (PV) and in some ET patients. TPO serum levels are not diagnostically useful and c-Mpl expression in megakaryocytes and platelets are generally decreased in ET. Mutations in TPO and c-MPL genes have been detected in familial thrombocythemia, but not in patients with acquired ET. The aim of the present study was to analyse PRV-1, TPO and c-MPL genes status by fluorescence in situ hybridization (FISH) technique in order to find new molecular markers in ET patients. Patients and Methods. Thirty bone marrow samples of ET patients (7M/23F) diagnozed by PVSG criteria with a normal karyotype and 10 bone marrow samples of normal healthy donors were included in the study. All samples were studied by three locus-specific probes for PRV-1, TPO and c-MPL genes as follows: 1. PRV-1 gene (BAC RP11-160A19, 157 Kb, located at 19q13.12-2) labeled in red cohybridized with the 19p telomeric probe (D19S238E, Vysis) labeled in green. 2. TPO gene (BAC RP11-45NP16, 183 Kb, located at 3q27) labeled in green cohybridized with the centromeric probe for chromosome 3 (D3Z1, Vysis) labeled in red. 3. c-MPL gene (BAC RP11-297L5, 190 Kb, located at 1p34) labeled in green cohybridized with the centromeric probe for chromosome 1 (D1Z5, Vysis) labeled in orange. A minimum of 100 interphase nuclei were analyzed. Results. FISH study showed no PRV-1, TPO and c-MPL cytogenetic abnormalities in any of the analyzed cases, except for one patient in which 21% of interphase nuclei presented a trisomy for the TPO gene region. The monosomy and trisomy thresholds were 6.1% and 4.7% for PRV-1, 4.9% and 3.4% for TPO, and 5.4% and 3.71% for c-MPL, respectively. Conclusions. Our results suggest a lack of structural and numerical rearrangements of PRV-1, TPO and c-MPL genes in ET patients. The PRV-1 gene FISH results are in line with the previously reported by Najfeld et al (Exp Hematol2003;31:118–21); regarding TPO and c-MPL results, this is the first FISH study reported in the literature in ET.

scholarly journals 5-bp Classical Satellite DNA Loci from Chromosome-1 Instability in Cervical Neoplasia Detected by DNA Breakage Detection/Fluorescence in Situ Hybridization (DBD-FISH)

2013 ◽  
Vol 14 (2) ◽  
pp. 4135-4147 ◽  
Author(s):  
Elva Cortés-Gutiérrez ◽  
Brenda Ortíz-Hernández ◽  
Martha Dávila-Rodríguez ◽  
Ricardo Cerda-Flores ◽  
José Fernández ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Satellite Dna ◽  
Cervical Neoplasia ◽  
Chromosome 1 ◽  
Dna Breakage
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