PubMed Health⌕ Search

Biomedical subjects

A V Carrano

Publications and source records attributed to A V Carrano.

At least 109 records · Page 6Linked to original sources

Chromosomal DNA cytophotometry in 20q- nonspecific myeloid disorders.

DNA cytophotometry was used to quantify the chromosomal alterations in the bone marrow and blood of three patients with nonspecific myeloid disorders. All patients possessed a population of cells with a morphologically abnormal chromosome 20, del(20)(qll). In two of the patients, the abnormal chromosome 20 showed nearly identical DNA measurements with a net loss of 0.37% of the total autosomal DNA in one patient and 0.38% in the second. The third patient had a net loss of only 0.25% of the autosomal DNA. Analysis of the DNA content of the long arm and short arm of the abnormal No. 20 indicated that all three cases had chromosomal material added to the short arm (0.10 to 0.14% of the autosomal DNA). About 0.50% of the autosomal DNA was deleted from the long arm in two of the patients; only 0.35% of the autosomal DNA was deleted from the long arm in the third case. Within the limit of resolution, there is no evidence that the material lost has been translocated intact to another chromosome. The origin of the 20q- chromosome as the result of an incomplete pericentric inversion is suggested.

Aged↗

Induction of long-lived chromosome damage, as manifested by sister-chromatid exchange, in lymphocytes of animals exposed to mitomycin-C.

The cytogenetic effects of repeated vs. acute exposure to a chemical mutagen--carcinogen were determined with an in vivo system in which chemicals injected into rabbits induce sister-chromatid exchanges (SCEs). SCE induction can be monitored when the animal's peripheral lymphocytes are cultured in the presence of bromodeoxyuridine (BrdUrd) and then scored for SCE frequency. Mitomycin-C (MMC), 0.5 mg/kg, was injected intraperitoneally once a week for 8 weeks. This treatment initially induced small increases in SCE frequency within one day of injection, followed by a return to control levels within 1 week. After the 4th injection, however, the frequency failed to return to normal. After the 5th injection, however it showed a 4-fold increase over the control which was sustained for the remaining 3 weeks of treatment and for an additional 2 weeks thereafter. The frequency then dropped to twice the control value and remained at this level for more than 4 months. All of the high SCE values after the first 4 weeks were due in part to the appearance and persistence of a population of cells with high SCE frequencies. Exposure to the same total dose given as a single injection resulted in a transient elevation in the SCE frequency and a subsequent return to lower values, with no evidence of a delayed effect such as the increase observed after 4 weeks in repeatedly exposed animals. Overall, repeated exposure is at least as effective as acute exposure in eliciting long-lived SCEs in vivo.

Animals↗

The distribution of mitomycin C-induced sister chromatid exchanges in the euchromatin and heterochromatin of the Indian muntjac.

The frequency of sister chromatid exchanges (SCEs) induced by mitomycin C (MMC) in Indian Muntjac chromosomes was determined by the fluorescence plus Giemsa (FPG) technique. Using scanning cytophotometry the relative DNA content of each chromosome was measured with and without acid or alkali pretreatments for C-banding. During acid and alkali treatments, euchromatin lost 20 to 30% of its DNA, while heterochromatin lost less than 5%; an intermediate DNA loss was observed for the short arm of the X chromosome. After growth of cells in the presence of MMC during the first cycle and in the presence of bromodeoxyuridine (BrdU) during the first and second cycles of DNA replication, SCEs in the euchromatin were proportional to DNA content. SCEs at the junctions between the neck of the X chromosome and the long and short arms occurred more frequently than expected. A threshold effect for the induction of SCEs was observed in regions resistant to DNA extraction by acid and alkali treatments (i.e., the neck and short arm of the X chromosome). At high concentrations of MMC, the frequency of SCE at each junction appears to plateau at 0.5.

Animals↗

The interaction of Hoechst 33258 and BrdU substituted DNA in the formation of sister chromatid exchanges.

Sister chromatid exchanges (SCEs) are induced in cultured Chinese hamster cells by treatment with 5-bromodeoxyuridine (BrdU) or with Hoechst 33258 (H33258) plus BrdU. The SCE frequencies depend upon the number of H33258 molecules available per cell (or per base pair) and the number of brdU molecules available per cell, and not solely upon molarity. In addition, H33258 and BrdU act synergistically to induce SCEs. At low BrdU concentrations H 33258 induces very few SCEs. At high BrdU concentrations and similar concentrations of H33258, however, SCE frequencies are significantly increased. SCE frequencies decrease with time in successively harvested cells because of the depletion of H33258 from the medium due to DNA binding.

Benzimidazoles↗

The DNA content of micronuclei induced in mouse bone marrow by gamma-irradiation: evidence that micronuclei arise from acentric chromosomal fragments.

The DNA contents of 75 micronuclei found in mouse bone marrow 48 h after a 260 R dose of gamma-rays were measured individually and compared to 71 diploid G1 nuclei. The coefficient of variation for the diploid cells was 6.3%. The DNA content of micronuclei varied from 0.5 to 11.1% of the diploid G1 nuclei with a mean of 3.5%. These results are in agreement with the expectations based upon the hypothesis that micronuclei arise from acentric chromosomal fragments produced by random breakage of the mouse genome.

Animals↗

DNA cytophotometry in pre-natal cytogenetic diagnosis.

DNA cytophotometry was used to resolve a cytogenetic ambiguity involving a possible deletion of part of the short arm of chromosome 18 in cells obtained by amniocentesis. Measurements of chromosomal DNA content were within normal limits. This is the first time that DNA cytophotometry has been used in cytogenetic diagnosis; it illustrates the potential of this new approach to cytogenetics.

Adult↗

Quantification by DNA-based cytophotometry of the 9q+/22q-chromosomal translocation associated with chronic myelogenous leukemia.

DNA-based cytophotometry was used to analyze metaphase chromosomes in four patients with chronic myelogenous leukemia. In three of these patients, both Philadelphia chromosome (Ph1)-positive and Ph1-negative cells were measured. On the basis of these three patients, the characteristic 9q+/22q- translocation of chronic myelogenous leukemia involves the net transfer of 0.325% of the autosomal genome; there is no evidence of net gain or loss of DNA (apart from duplication of the Ph1 chromosome in one patient), and no significant difference is found in the amount of DNA transferred in different patients. Significant differences are found among patients in the derived Chromosomes 9 and the Ph1 chromosomes and are ascribed to preexisting variations in the Ph1-negative cells of these patients. There is no evidence in these patients of any further cytogenetic lesion associated with chronic myelogenous leukemia.

Adolescent↗

Purification of the chromosomes of the Indian muntjac by flow sorting.

Metaphase chromosomes were isolated from a male Indian muntjac cell line, were stained with ethidium bromide and were analyzed by flow microfluorometry to establish a deoxyribonucleic acid (DNA)-based karyotype. Five major peaks were evident on the chromosomal DNA distribution corresponding to the five chromosome types in this species. The amount of DNA in each chromosome was confirmed by cytophotometric measurements of intact metaphase spreads. The five chromosome types were separated by flow sorting at rates up to several hundred chromosomes per second. The sorted chromosomes were identified by morphology and by Giemsa banding patterns. The automsomes, Numbers 1, 2 and 3, and the X + 3 composite chromosome were separated with a high degree of purity (90%). The centromere region of the X + 3 chromosome was fragile to mechanical shearing, and during isolation a small proportion of these chromosomes broke into four segiments: the long arm, the short arm, the short arm plus centromere and the centromere region. A large fraction of the constitutive heterochromatin of this species is present in the centromere region of the X + 3 chromosome and in the Y chromosome; these two regions possess similar amounts of DNA and therefore sort together. Chromosome flow sorting is rapid, reproducible and precise; it allows the collection of microgram quantities of purified chromosomes.

Animals↗

Distribution of sister chromatid exchanges in the euchromatin and heterochromatin of the Indian muntjac.

The frequency of sister chromatid exchanges (SCEs) was determined for the chromosomes (except Y2) of the Indian muntjac stained by the fluorescence plus Giemsa (FPG) or harlequin chromosome technique. The relative DNA content of each of the chromosomes was also measured by scanning cytophotometry. After growth in bromodeoxyuridine (BrdU) for two DNA replication cycles. SCEs were distributed according to the Poisson formula in each of the chromosomes. The frequency of SCE in each of the chromosomes was directly proportional to DNA content. A more detailed analysis of SCEs was performed for the three morphologically distinguishable regions of the X-autosome composite chromosome. The SCE frequency in the euchromatic long arm and short arm were proportional to the amount of DNA. In contrast, the constitutive heterochromatin in the neck of this chromosome contained far fewer SCEs than expected on the basis of the amount of DNA in this region. A high frequency of SCE, however, was observed at the point junctions between the euchromatin and heterochromatin.

Animals↗

Chromosome measurement and sorting by flow systems.

A flow microfluorometer was used to measure metaphase chromosomes in suspension at rates up to 100,000 per min. Chromosomes from cells of the Chinese hamster M3-1 cell line were isolated, stained for DNA with the fluorescent dye ethidium bromide, and analyzed for DNA content. Nine distinct peaks were resolved that correspond well with independent chromosomal DNA measurements made with a high-resolution scanning cytophotometer. Chromosomes were sorted from each peak by an electronic cell sorter. Visual examination of each fraction indicated the purity of the sorted chromosomes. This novel technology allows separation of purified populations of individual chromosomes suitable for biochemical and biological characterizations.

Animals↗

High-speed quantitative karyotyping by flow microfluorometry.

We describe a new flow-system technique for high-speed quantitative karyotyping. Metaphase chromosomes are isolated from cells, stained with a DNA-specific fluorochrome, and individually measured for stain content at rates up to 10(5)/min in a flow microfluorometer. The resulting distribution contains quantitative information about the relative frequency of occurrence and the chromosomal DNA content of the measured chromosomes. Such analyses of Chinese hamster chromosomes are in good agreement with independent results from scanning cytophotometry and with cytogenetic observations of the banded chromosomes. Analyses of two karyotypically different clones of Chinese hamseter M3-1 lines provide the quantitative characterization of two stable translocations. Initial data on human chromosomes are presented and agree well with results of scanning cytophotometry and suggest potential clinical applications as preparative and instrumental techniques improve.

Animals↗