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PubMed2000

[Dynamic interphasic tensimetry of blood and urine in systemic lupus erythemotosus].

AIM: To study dynamic surface tension (DST) of blood and urine in SLE patients. MATERIALS AND METHODS: Clinical, immunological examinations, DST of blood and urine using computer tensiometers were made in 67 SLE patients aged 15 to 62 years. RESULTS: Changes in interphasic tensiograms in males and females were characterized regarding the course, activity of the pathological process and the presence of certain clinical symptoms. Effective therapy return to normal DST of the biological fluids. Correlations between interphasic tensiograms of blood, urine and levels of proteins, lipids, non-protein and inorganic surfactants and surface-inactive substances were established as well as relations of DST with biochemical composition of the biological fluids. CONCLUSION: The above diagnostic method may be useful for differential diagnosis, assessment of the clinical course, activity of the disease, control over effectiveness of the treatment.

Adolescent↗
PubMed2001

Detection of trisomy 8 in philadelphia chromosome-positive CML patients using conventional cytogenetic and interphase fluorescence in situ hybridization techniques and its relation to c-myc involvement.

Trisomy 8 (+8) is a common clonal evolution marker for progression in chronic myelogenous leukemia. The relationship of +8 to various stages of t(9;22) leukemias is not firmly established. To explore this association we examined bone marrow (BM) cells from 10 Philadelphia chromosome positive (Ph+) chronic myeloid leukemia (CML) patients in different stages of the disease, using conventional cytogenetic technique(CCT) and interphase fluorescence in situ hybridization (FISH). FISH detection of chromosome 8 was accomplished using the D8Z2 (Oncor) probe specific for the centrometric region of chromosome 8. Five hundred interphase nuclei were counted for each patient. Three of the 10 patients were selected for detection of c-myc gene locus located in the 8q24.2-24.3 region using the L

Adult↗
PubMed1975

RNA synthesis in Chinese hamster cells. III. Non-coordinate increases during interphase in synthesis rates for informosomal, polysomal and heterogeneous nuclear RNAs.

Cultured Chinese hamster ovary cells were synchronized by mitotic selection. Relative synthesis rates for informosomal messenger-like RNA (mlRNA), polysomal messenger RNA (mRNA), and heterogeneous nuclear RNA (HnRNA) were estimated from the amount of labeled adenosine or uridine incorporated into these species in early and late interphase. The amounts of uridine incorporated into HnRNA, mRNA, and mlRNA during a pulse administered 9.75-10.75 h post-mitosis were 3.48 4.64, and 2.82 times the amounts incorporated 1.5-2.5 h post-mitosis. Adenosine incorporation values 9.5-11.0 h post-mitosis were 1.64 (HnRNA), 2.49 (mRNA), and 1.18 (mlRNA) times the 1.5-3.0 h values. The realitive incorporation into MRNA of large polysomes corresponded to incorporation into mRNA of smell polysomes. Thus, the synthesis rates of mRNA, mlRNA, and HnRNA increase during interphase in a noncoordinate fashion.

Adenosine↗
PubMed2001

[Interphase FISH analysis of frozen or fixed tissues for the detection of t(11;14) (q13;q32) in mantle cell lymphoma].

Cytogenetic analyses have revealed that mantle cell lymphomas (MCL) are closely associated with the t(11;14)(q13;q32). This translocation juxtaposes the immunoglobulin heavy chain gene (IGH) sequences with the BCL-1 locus, leading to up-regulation of the CCND1 gene and consequently to an overexpression of cyclin D1 protein. We studied 27 MCL with characteristic morphological and immunological (CD5+, CD10-, CD20+, CD23-) features and 2 controls (reactionnal lymphadenitis) to evaluate the feasibility and the interest of FISH analysis on interphase cells from frozen or paraffin-embedded tissues. Sections (CC) and touch preparations (EC) of frozen tissues and sections of paraffin-embedded tissues (CF) were successfully hybridized with the Vysis LSI IgH/CCND1 dual color dual fusion translocation probe. The touch preparations presented a lower cellularity than sections, therefore allowing an easier analysis. Hybridization spots intensities were found stronger in CC and EC than in CF. The percentages of t(11;14) positive cells were similar in CC, EC and CF from a same patient. The percentage of non hybridized cells, analogous in CC and EC, was higher in CF. However, the CF were directly analysed on microscope without the need of any numerical picture treatment. The t(11;14) was detected in all the cases (27/27) and positive cells percentages were always higher than the probe cut-off (5%). The FISH analysis on interphase cells appears a performing and rapid technique to detect t(11;14) in MCL on both frozen and paraffin-embedded tissue, thus extending its practical and diagnostic use.

Cyclin D1↗
PubMed2002

[Interphase FISH analysis on histologic sections of fixed tissues for t(11;14) (q13;q32) detection in mantle cell lymphoma and t(8;14)(q24;q32) in Burkitt's lymphoma].

We describe an interphase FISH analysis for formol-fixed, paraffin-embedded tissue sections with commercial probes detecting the t(11;14)(q13;q32) in mantle cell lymphoma and the t(8;14)(q24;q32) in Burkitt's lymphoma. Staining of an adjacent section allowed identification of tumoral areas. The cut-off value was evaluated in reactive lymph nodes at 5% for both probes. An incomplete hybridization pattern was found in 18 to 26% of the nuclei but was always lower than the percentages of translocated cells in tumoral regions. A t(11;14) was detected in 4/4 mantle cell lymphomas and a t(8;14) in 2/3 Burkitt's lymphomas. Interphase FISH analysis of fixed-tissue sections is a reliable technique for the direct detection of lymphoma-associated translocations on routine histological material.

Burkitt Lymphoma↗
PubMed1992

Simultaneous genotypic and immunophenotypic analysis of interphase cells using dual-color fluorescence: a demonstration of lineage involvement in polycythemia vera.

Fluorescent in situ hybridization has become a useful technique by which chromosomal abnormalities may be shown in interphase cells. We present a dual-fluorescence method whereby a chromosomal and immunophenotypic marker can be visualized simultaneously in the same interphase cell. Two patients with the myeloproliferative disorder polycythemia vera and trisomy for chromosome 8 have been studied using this technique and selective involvement of the myeloid and erythrocyte lineages has been shown by the detection of the trisomy in immunophenotyped cells. Simultaneous analysis of genotype and immunophenotype in individual cells from patients with myeloproliferative disorders or leukemia may help identify the developmental and lineage status of cells in which molecular alterations have resulted in clonal advantage.

Aged↗
PubMed1992

Structural chromosome 1 aberrations in transitional cell carcinoma of the bladder: interphase cytogenetics combining a centromeric, telomeric, and library DNA probe.

Fluorescence in situ hybridization (FISH) was used to study numerical and structural chromosome 1 aberrations in interphase nuclei of transitional cell carcinomas (TCCs) of the urinary bladder. One of the characteristic numerical aberrations, as detected previously in low-grade noninvasive TCCs, included trisomy for chromosome 1 (A. H. N. Hopman et al., Cancer Res., 51: 644-651, 1991). We examined in more detail 22 cases with a centromeric (1q12) and a telomeric associated (1p36) DNA probe and with a library DNA probe from sorted human chromosome 1 in single- and double-target FISH procedures. All flow cytometrically determined DNA diploid TCCs (13 cases), which showed three spots for 1q12 (6 cases), had two spots for 1p36. Since the library DNA probe showed three separate domains in the nuclei of these cases, the additional copy for 1q12 could be explained as an extra chromosome 1p-, containing the 1q12 target. In the flow cytometrically determined DNA tetraploid/aneuploid tumors, the results were more complex. In 6 of 9 cases, we observed an overrepresentation of 1q12 as compared to 1p36, also suggesting the presence of extra copies of 1p- chromosomes. The results of the present study demonstrate the utility of the FISH method to assess structural chromosome aberrations in interphase nuclei of solid tumors.

Carcinoma, Transitional Cell↗
PubMed1992

Detection of trisomy 12 in chronic lymphocytic leukemia by fluorescence in situ hybridization to interphase cells: a simple and sensitive method.

Trisomy 12 is the most common cytogenetic abnormality in chronic lymphocytic leukemia (CLL), and a number of studies have suggested that it may be an adverse prognostic indicator. We have evaluated the usefulness of fluorescence in situ hybridization with a chromosome 12-specific probe as a simple means for detecting trisomy 12 in interphase cells. Forty cases of B-cell CLL previously studied with conventional cytogenetic techniques were analyzed with a biotinylated probe to the centromeric region of chromosome 12. Thirty of these retrospective cases could be reevaluated with in situ hybridization. Our analysis showed three hybridization signals (ie, trisomy 12) in interphase cells from seven of seven cases found previously to have trisomy 12. Trisomy 12 was also detected in five additional cases: in one case thought to have a normal karyotype, in two cases that had been inadequate for routine cytogenetic analysis, and in two cases that had been found to have an abnormal karyotype without trisomy 12. In a prospective series of 20 newly accrued CLL cases, all cases were analyzed successfully by in situ hybridization and six (30%) showed trisomy 12. We were able to perform the analysis on routinely prepared and previously Wright-stained peripheral blood smears. We conclude that fluorescence in situ hybridization is a simple means for the detection of trisomy 12 in CLL. The technique is more sensitive than conventional cytogenetic analysis and would be a useful tool in clinical studies.

Chromosomes, Human, Pair 12↗
PubMed2005

Chromosomal instability detected by interphase fluorescence in situ hybridization and its relation to p53 alteration in prostate carcinoma in Saudi patients.

OBJECTIVE: Chromosomal instability (CIN) is a feature of human neoplasm. The p53 mutation has been shown to be associated with CIN in many human dysplastic and neoplastic lesions. The objective of this study was to examine CIN and p53 mutations in prostate carcinoma (Pca) resected from Saudi patients. METHODS: Testing of p53 alteration using immunohistochemistry was performed on 28 archived prostatic carcinoma specimens containing Pca foci from Saudi patients seen at King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia. Chromosomal instability was evaluated in the same tissues by interphase in situ hybridization (IFISH) using centromere probes for chromosome 7 and 8. Immunohistochemistry and IFISH were performed at Princess Margaret Hospital, University Health Network, Toronto, Ontario, Canada in 2001. RESULTS: The p53 immunoreactivity was found in 29% in Pca and 0% in benign epithelium. Interphase in situ hybridization revealed numerical chromosomal alterations in keeping with CIN in 63% of p53 positive and 20% p53 negative Pca. No evidence of CIN was seen in non-neoplastic epithelium. CONCLUSION: We concluded that CIN as determined by IFISH is present in Pca from Saudi patients similarly to those reported in western countries. The p53 mutation occurs relatively infrequently in Pca and is associated with the presence of CIN at least in a subset of Pca.

Chromosomal Instability↗
PubMed2005

Detection of 22 q11.2 hemizygous deletion by interphase FISH in a patient with features of CATCH 22 syndrome.

We report an eight years female child with clinical and molecular cytogenetic findings consistent with CATCH 22 syndrome characterized by cardiac defect, typical facial dysmorphism, mental deficiency and chromosome 22 q11.2 deletion. Interphase FISH with 22q 11.2 probe demonstrated hemizygous deletion in 98.5% nuclei. Interphase FISH for diagnosis of CATCH 22 syndrome has not been reported previously from India to our knowledge.

Abnormalities, Multiple↗
PubMed2006

Split centromeric alpha-satellite FISH signals in a whole arm translocation 5p;10p: consequences and implications for interphase FISH studies.

This is a case report of an apparently balanced whole arm translocation between the short arms of chromosomes 5 and 10 in which the centromeric alpha-satellite DNA is split between both derivative chromosomes for both probes, leading to abnormal signal patterns. The patient requested preimplantation genetic testing for the unbalanced products of the translocation. However, using centromeric alpha-satellite DNA probes as controls for the subtelomeric-specific probes in interphase was not informative because of the split signals. The ramifications of such variation in the alpha-satellite regions of chromosomes for other interphase FISH tests are discussed.

Journal Article↗
PubMed2006

[Monitoring of bcr/abl fusion gene by interphase-dual-color and dual-fusion fluorescence in situ hybridization in CML after allo-HSCT].

This study was aimed to investigate the sensitivity and clinical application value of interphase-dual-color and dual-fusion fluorescence in situ hybridization (DD-FISH). The minimal residual disease (MRD) in 19 patients with chronic myelogenous leukemia (CML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) was detected by DD-FISH, and the detected results were compared with those of conventional cytogenetics (CC) and reverse transcription-polymerase chain reaction (RT-PCR). The samples were collected from bone marrow or peripheral blood or smears of bone marrow. The results indicated that 14 out of 19 patients achieved and maintained continuous complete molecular remission after transplantation. In these patients, CC assay displayed normal donor karyotype, result of RT-PCR was negative, complete donor chimerism was detected after 2 months of transplantation, result of DD-FISH was negative, average time of the follow-up survey was 11.25 months, MRD did not increase. Results of CC and RT-PCR in 1 patient showed negative, while FISH of sex chromosome showed mixed chimerism, result of DD-FISH was positive, MRD did not increase, no therapy was given for this patient, clinical state of patient was stable. Three patients with hematological relapse demonstrated obvious increase of MRD detected by DD-FISH and sex FISH, result of RT-PCR was found positive in them, but the abnormal result of CC was observed only in 1 patient. After donor lymphocyte infusion and imatinib mesylate treatment, these 3 patients achieved cytogenetic remission again, results of DD-FISH, CC and PCR were negative in them. DD-FISH, CC and PCR in bone marrow and peripheral blood from one patient with extramedullary relapse revealed negative results, and the complete chimerism was found in this patient. It is concluded that interphase-dual-color and dual-fusion fluorescence in situ hybridization is a more reliably sensitive and practicable method for monitoring MRD in patients with CML after allo-HSCT, and can be used in detection of chromosome sample and blood or bone marrow smears. Dynamic detection of bcr/abl fusion gene level by FISH may predict disease changes and guide individual therapy.

Adolescent↗
PubMed1977

[Electron microscopic study of interphase in a culture of fibroblast-like cells].

The analysis of the ultrastructural changes of fibroblast-like cells of Chinese hamster at different periods of the cell cycle demonstrates that the most reliable morphological criteria for identification of different periods of the interphase are changes of the nuclear chromatin and nucleolus and the dynamics of development of the daughter centrioles. No prominent changes of the morphology of the protein-synthesizing apparatus during interphase were noted.

Animals↗
PubMed1981

Disappearance of microvilli accompanying radiation-induced interphase death of rat thymocytes.

Thymocytes are radiosensitive and show interphase death (immediate death) after irradiation. The present investigation was undertaken to characterize by scanning electron microscopy the shape of thymocytes irradiated in vitro with 1 kR X-rays and determine the cell surface changes that accompany the development of interphase death. Exposing the cells to X-rays caused disappearance of the microvilli, usually observed with viable cells. Association of dead thymocytes with porous surface without any digitations was confirmed by the examination of enriched dead cell populations prepared by centrifuging the cell suspension in Percoll density gradient.

Animals↗
PubMed1995

[The effect of certain factors on binding of tert-butyloxycarbonylamino acids to Merrifield polymer during interphase catalysis and assessment of racemization of the bound amino acid].

In order to reduce the influence of hydrogen bonds on the acylamino acid salts attachment to the chloromethylated resin, it is proposed to use compounds that can compete for the hydrogen bonds formation. The best solvent proved to be hexamethylphosphoric triamide. Use of interphase catalysts, e.g., tributyl-p-nitrobenzyl ammonium, also gives good results. The racemization degree of the amino acids attached to solid support by means of the interphase catalysis does not exceed that of amino acids loaded on the polymer according to Gisin's method.

Amino Acid Sequence↗
PubMed1993

Interphase in situ hybridization reveals minimal residual disease in early remission and return of the diagnostic clone in karyotypically normal relapse of acute lymphoblastic leukemia.

Early clinical remission of acute lymphoblastic leukemia (ALL) was examined by fluorescence in situ hybridization (FISH) in bone marrow cells from eight patients with high hyperdiploid (> 50 chromosomes) clones at diagnosis. Alphoid centromeric probes to chromosomes X, 10, 17, and 18, trisomic at diagnosis, were used as appropriate. Three hematologically normal marrows were used as controls. At diagnosis, trisomic interphase cells ranged from 69.3-84.4%. One month later, trisomic and tetrasomic interphase cells were significantly increased over control frequencies in 2/7 cases tested (p < 0.05 and p < 0.001) and trisomy alone in one case (p < 0.05). At two months post-diagnosis, trisomy and tetrasomy were in the control range. Pentasomy and hexasomy, not seen in controls, were found in 5/7 samples at one month and in 1/5 samples at two months. One patient examined in chromosomally normal relapse had 34.4% trisomic cells by FISH at confirmed relapse (p < 0.001). An additional hyperdiploid case, examined at central nervous system relapse, had increased numbers of trisomic cells (p < 0.01) in a morphologically and cytogenetically normal marrow. These findings demonstrate the elimination of aberrant ploidy in most hyperdiploid cases two months from diagnosis and indicate that failure to detect the abnormal clone in relapse may result from selective mitotic activity of chromosomally normal cells, rather than relapse in a cytogenetically normal clone.

Adult↗
PubMed1996

Persistent gestational trophoblastic disease: DNA image cytometry and interphase cytogenetics have limited predictive value.

DNA flow cytometry has shown a wider spectrum of DNA content in the complete hydatidiform mole (CM) than the originally reported diploidy. Conflicting results have been published about the relationship of DNA content and the occurrence of persistent gestational trophoblastic disease (PGTD). In the present study, 71 cases of CM and 4 cases of partial mole accompanied by PGTD and 100 cases of CM without PGTD were evaluated with DNA image cytometry for differences in DNA-ploidy pattern, expressed as the 2.5c and 5c exceeding rates. A pilot study of 20 cases of each group was performed using interphase cytogenetics to detect differences in the frequency of numerical chromosomal aberrations and in sex chromosome composition. For this purpose, DNA probes specific for the pericentromeric regions of chromosomes 1 and X and for the long arm of chromosome Y were incubated on 6-micron paraffin tissue sections. The results showed no differences between CMs with or without PGTD; DNA polyploidy occurred in 99% and 98% of cases, respectively; the 2.5c exceeding rate and 5c exceeding rate were 62.6 and 62.4, and 6.5 and 6.0, respectively. The frequency of numerical chromosomal aberrations as detected by interphase cytogenetics was 23.4 and 22.8%. An XY pattern was found in 3 of 20 cases of CM with PGTD and in 4 of 20 cases of CM without PGTD. The four cases of partial mole showed a DNA-ploidy pattern identical to that of a CM. For this reason, they would be better reclassified as CMs, despite the presence of nucleated red blood cells or amnion. Although nuclear atypia and corresponding increased DNA content is pronounced but variable in CMs, the occurrence of PGTD is not related to variations in quantitative DNA content nor to gross heterology or homology in sex chromosomes.

Adult↗
PubMed1996

[Detection of microbubbles formed in the blood/CO2 interphase during decompression in barometric conditions similar to laparoscopy].

HYPOTHESIS AND OBJECTIVES: Intraperitoneal insufflation (IPI) with CO2 during laparoscopic surgery establishes a pressure gradient that determines the passage of gas from the peritoneal cavity to the blood and surrounding tissues. The transport and clearance of CO2 are assured by proper sweeping when regional blood flow is adequate in volume and distribution. But if IPI hyperpressure surpasses regional venous capillary pressure (10 to 15 mmHg) and there is no cardio-circulatory adaptation to the phenomenon, CO2 clearance may be compromised. Under these conditions, the expected post-insufflation increase in PetCO2 will not take place. Bearing in mind the physical characteristics of CO2, retention of this gas in the intraperitoneal cavity produces blood and tissue saturation under a higher-than-atmospheric pressure, after a certain period of time in contact. Rapid intraperitoneal decompression after laparoscopic surgery carries with it the risk of microbubble formation due to release of CO2 that had been dissolved under hyperbaric conditions. MATERIAL AND METHODS: To test this hypothesis, the barometric conditions of laparoscopy were reproduced inside an observation capsule containing blood and CO2. RESULTS: Magnification revealed that after decompression bubbles formed in the blood/CO2 interphase. The images were recorded on magnetic videotape. Thirty minutes after decompression, the bubbles could still be seen, even after the interphase was swept with a current of air. DISCUSSION: Rapid intraperitoneal decompression after laparoscopy can generate the formation of microbubbles which, if not eliminated, will give rise to local ischemic manifestations. This same decompression, correcting the local circulatory alterations and activating the CO2 transport that had been compromised, could introduce gas bubbles into the blood stream such as are responsible for delayed gaseous microembolism. The simultaneous observation of changes in PetCO2 (stability or post-insufflation decreases) and hemodynamic parameters during laparoscopy, would allow evolving anomalies to be detected early and therapeutic action to be taken to prevent the formation of microbubbles.

Carbon Dioxide↗