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Biomedical subjects

M E Drets

Publications and source records attributed to M E Drets.

At least 19 recordsLinked to original sources

Cytological indications of the complex subtelomeric structure.

Research on the subtelomeric region has considerably increased because this chromosome segment (1) keeps the chromosome number constant, (2) intervenes in cancer and cell senescence processes, (3) presents more crossovers than other regions of the genome and, (4) is the site of cryptic chromosome aberrations associated with mental retardation and congenital malformations. Quantitative microphotometrical scanning and computer graphic image analysis enables the detection of differentially distributed Giemsa-stained structures in T-banded subtelomeric segments of human and Chinese hamster ovary (CHO) chromosomes. The presence of high density stain patterns in the subtelomeric region was confirmed using endoreduplicated chromosomes as a model. Besides, prolonging the incubation in the T-buffer, specific holes were induced in subtelomeric segments. Hole specificity was confirmed inducing them in complex CHO chromosome aberrations obtained by AluI. The method was also used to detect minute sister chromatid exchanges in the T-banded subtelomeric area (t-SCEs). The presence of t-SCEs was suspected to reflect, at the microscope level, the high crossover activity prevailing in the region. Due to the fact that the fluorescent signals obtained with subtelomeric probes seem to be colocalized with subtelomeric high density areas, measurements on the position of both structures with respect to the diffraction and chromosome edges were carried out. Data obtained showed comparable values suggesting that the high density segments were located where telomeric probes usually fluoresce. The possible relationship of the high density patterns, the production of specific holes, the localization of fluorescent areas and the detection of minute SCEs in the subtelomeric segment observed in T-banded CHO and human chromosomes is briefly reviewed.

Animals↗

Chromosomal aberrations: formation, identification and distribution.

Chromosomal aberrations (CA) are the microscopically visible part of a wide spectrum of DNA changes generated by different repair mechanisms of DNA double strand breaks (DSB). The method of fluorescence in situ hybridisation (FISH) has uncovered unexpected complexities of CA and this will lead to changes in our thinking about the origin of CA. The inter- and intrachromosomal distribution of breakpoints is generally not random. CA breakpoints occur preferentially in active chromatin. Deviations from expected interchromosomal distributions of breakpoints may result from the arrangement of chromosomes in the interphase nucleus and/or from different sensitivities of chromosomes with respect to the formation of CA. Telomeres and interstitial telomere repeat like sequences play an important role in the formation of CA. Subtelomeric regions are hot spots for the formation of symmetrical exchanges between homologous chromatids and cryptic aberrations in these regions are associated with human congenital abnormalities.

Animals↗

The underlying structure of the subtelomeric region detected by microphotometrical scanning and chromosome graphic image analysis.

Previous research showed that the microphotometrical scanning of T-banded subtelomeric regions reveals the presence of specific patterns of the Giemsa stain density distributions as detected in chromosomes of normal human lymphocytes and CHO cells. Analyses with this method of the T-banded subtelomeric segments of CHO endoreduplicated chromosomes confirmed that these density patterns replicate in a similar way in sister chromosomes. Besides, the specific removal of portions of the subtelomeric segments appearing as tiny holes located where these density patterns are found suggested that both phenomena are related. The possible connection of these findings to the molecular structure of the subtelomeric region is briefly discussed.

Animals↗

Computer graphics as a tool in cytogenetic research and education.

Scanning microscope photometry has been extensively used for image analysis of nuclei and chromosomes and for automated karyotyping. Color graphics terminals, software development and appropriate data manipulation have expanded the scope of scanning microphotometry so that it is now particularly useful for the quantitative analysis of cell components. We have developed application software for displaying nuclear and chromosome densities on a computer terminal of in the form of pixeled images and pseudo-three-dimensional graphic diagrams, as well as for image transformation, object illumination and rotation to enhance chromatin features. The system allowed the microphotometrical scanning of human G-, R-, C-, T-banded chromosomes and sister chromatid exchanges, the measurement of nuclear heterochromatic segments, the detection of minute chromosome structures and the distribution of high staining densities in chromosomes which are difficult to observe with simple light microscopy. The new quantitative visual information given by our interactive graphic method contributes to a better understanding of the nuclear and chromosome structure for the students engaged in cytogenetic research and teaching. The analytical perspective of computer graphics in cytogenetic research and education is briefly discussed.

Animals↗

Clastogenic action of a dimethyl p-benzoquinone of animal origin.

The cytogenetic action of a volatile dimethyl p-benzoquinone found in the natural secretion of an arachnid (Acanthopachylus aculeatus Kirby) was studied in cultured human peripheral leukocytes and in mouse bone-marrow cells. Continuous and pulse treatments carried out in vitro, as well as experiments performed in vivo, induced different chromatid and chromosome aberrations suggesting that this chemical has clastogenic properties. The biology of the animal and the possible role of one of the components of its secretion as a natural mutagenic agent are discussed.

Animals↗

Chromosome delineation induced by a combined potassium permanganate-sodium bisulfite treatment.

A cytological procedure for in vitro chromosome delineation has been studied using human and mouse (Mus musculus) chromosomes. This method, consisting of slide incubation in KMnO4 at 0--5 degrees C for 24 h followed by a short exposure to NaHSO3 (1--3 min) and Giemsa staining, induces extraction of chromatin from human and mouse interphase nuclei and chromosomes. Autoradiography after 3H-ThD incorporation in vitro and cytophotometry confirmed that DNA is removed. Well contour-delineated and non-distorted chromosomes are observed in both species allowing the identification of all human chromosome groups. Contour chromosome delineation and its relationship to chromosome organization is briefly discussed.

Animals↗

BANDSCAN--a computer program for on-line linear scanning of human banded chromosomes.

BANDSCAN, an interactive program for on-line linear scanning of human G-banded chromosomes quantitative analysis is described. This program was written for a Wang 720 C programmable desk calculator associated to the Zeiss scanning photometer MP01. The system can detect up to a maximum of 24 densitometric band peaks found along banded chromosome arms or chromatids, estimate the total arm length and localize bands in terms of their relative positions. The scanning stage under control is always returned to the pre-fixed scanning starting point (centromere) which allows a user to scan the specimen repeatedly at different sensitivities and thus to reject minor bands or suspected chromosome artifacts. This facilitates a better visualization of major bands and chromosome landmarks. A fully formated print-out on band localizations and their relative positions is obtained at the end of each scanning. The possibilities of the application of this program to band mapping of human chromosomes and to the study of small chromosome band aberrations is discussed.

Animals↗

Aanlysis of the frequency and distribution of sister chromatid exchanges in cultured human lymphocytes.

Lymphocytes from 20 notmal subjects (11 male and 9 female) were examined for the frequency and location of sister chromatid exchanges (SCE) by the BrdU--Giemsa method. The mean frequency of SCE was 6.37 with little significant variation. One subject had a high number of exchanges in chromosome 1 while the remainder showed a random distribution of exchanges between chromosomes. The frequency of exchanges generally increased with chromosome length. However, chromosome 1, 2 and the B group had more exchanges than expected while the E, F and G grous had less than expected. The distribution of exchanges in chromosomes 1, 2 and the B group was non-random with a concentration of exchanges below the centromere and to a lesser extent on the distal portion of the long arm. The majority of exchanges appeared to occur at the junction between the dark and light G bands. It is suggested that the concentration of exchanges may reflect differences in BrdU incorporation along the length of the chromosome.

Bromodeoxyuridine↗

Mechanisms of chromosome banding. IX. Are variations in DNA base composition adequate to account for quinacrine, Hoechst 33258 and daunomycin banding?

Prior studies on subfractions of mouse and Kangaroo rat DNA have suggested that variations in base concentration within a given genome may not be great enough to account for Q-banding. To examine this with another species, calf DNA was subfractionated by CsCl ultracentrifugation into GC-rich satellites and the main band DNA was further fractionated into AT-rich, intermediate and GC-rich portions. The effect of varying concentrations of these DNAs on quinacrine and Hoechst 33258 fluorescence was examined. Although with both compounds there was less fluorescence in the presence of the GC-rich satellites than main band fractions, these results per se did not answer the question of whether the variation in base composition alone was adequate to account for chromosome banding. To answer this the fluorescence observed in the presence of DNA of a given base composition was related to the fluorescence observed in the presence of DNA of 40% GC content (F/F40). This allowed the derivation of a term B which indicated the relative change in fluorescence per 1% change in base composition of DNA. To determine the percent change in fluorescence observed in Q-banding, the photoelectric recordings of Caspersson et al. (1971) were used. From these data we conclude: 1. Quinacrine is twice as sensitive to changes in base composition as Hoechst 33258. 2. Variation in the base content of DNA along the base content of DNA along the chromosome is sufficient to account for most Q-banding, except possibly for some of the extremes of quinacrine fluorescence. This was further examined with daunomycin. Even though daunomycin gives good fluorescent banding, DNAs varying in base composition from 100 to 40% GC content all resulted in the same relative fluorescence of 0.03. However, in the presence of poly (dA-dT) the relative fluorescence was 0.85, indicating a great sensitivity to very AT-rich DNA. This suggests that with daunomycin and possibly other fluorochromes, stretches of very AT-rich DNA may be more important in fluorescent banding than simple variation in mean base composition.

Adenine Nucleotides↗

Specific banding patterns of human chromosomes.

Individual pairs of human chromosomes can be reliably identified by a new method that does not require special optical equipment and that results in permanent preparations. This method, which is based on treatment of the chromosomes in situ with NaOH, followed by incubation in sodium chloride-trisodium citrate and Giemsa staining, results in highly specific banding patterns in characteristic regions of the chromosome arms. It should prove useful for the detection of small structural changes in chromosomes.

Chromosomes↗