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Biomedical subjects

M D Bennett

Publications and source records attributed to M D Bennett.

At least 19 recordsLinked to original sources
PubMed1995

Chromosomal location of endogenous geminivirus-related DNA sequences in Nicotiana tabacum L.

The N. tabacum (tobacco) nuclear genome carries approximately 25 multiple direct repeats of a geminivirus-related DNA (GRD) sequence that probably arose by illegitimate recombination, following geminivirus infection, during Nicotiana evolution. Each GRD repeat carries sequences similar to the geminiviral AL1 gene of the tomato golden mosaic virus (TGMV), encoding a protein required for viral DNA replication, plus the cis-essential replication origin. Using a cloned 14-kb GRD repeat sequence as a probe for fluorescence in situ hybridization (FISH), we identified a unique tobacco chromosome carrying GRD. Translocations between chromosomes of the tobacco S and T genomes were used as physical markers by sequentially hybridizing chromosomes with labelled GRD and total genomic DNA from N. sylvestris (equivalent to the S genome). The 25S, 18S and 5.8S ribosomal gene clusters were detected in double-labelling experiments for use as additional markers to identify the chromosomal location of GRD. GRD occupies one site on a homologous pair of small submetacentrics from the T genome characterized by a lack of either translocated segments from the S genome or ribosomal genes. GRD provides an additional marker for the small chromosomes of the T genome and a useful phylogenetic tool.

Chromosome Mapping↗
PubMed1995

Chromosome identification and mapping in the grass Zingeria biebersteiniana (2n = 4) using fluorochromes.

The grass Zingeria biebersteiniana is one of five angiosperms known with 2n = 2x = 4. Its chromosomes were studied using fluorochrome banding and fluorescence in situ hybridization (FISH). The large pericentromeric region fluoresced much more brightly on chromosome 2 than on chromosome 1, using two different fluorochrome banding methods. These offer rapid and reliable means for identifying chromosomes and work throughout mitosis. FISH located the major site of 18S-26S rDNA sequences at the secondary constriction, which is proximal to two minor sites, all on the short arm of chromosome 1. Two 5S sites were also detected, the most distinct on the short arm of chromosome 2 and the other apparently co-localized with part of the major 18S-26S rDNA cluster on chromosome 1. These results constitute the first steps in constructing a physical gene map for Z. biebersteiniana. Such information may facilitate future studies of the organization and reorganization of grass genomes, including research into the spatial arrangement of the genome in Zingeria nuclei and much wider comparisons of synteny and genome evolution in grasses.

Centromere↗
PubMed1995

Evidence for somatic translocation during potato dihaploid induction.

Potato dihaploid PDH55 (Solanum tuberosum) is exclusively euploid (2n = 24) but apparently contains and expresses DNA from dihaploid inducer IVP48 (S. phureja). Genomic in situ hybridization (GISH) suggested IVP48 DNA incorporated stably into PDH55 by somatic translocation. This finding has two important implications. Firstly, the long-held implicit assumption that euploid dihaploids produced by dihaploid inducers are pure S. tuberosum seems incorrect. This may complicate meiotic, genetical and molecular studies involving potato dihaploids. Secondly, if such translocations are not rare, the phenomenon may offer a novel way to introduce useful traits directly from wild dihaploid-inducing species into S. tuberosum.

Base Sequence↗
PubMed1992

Genome reorganization in Nicotiana asymmetric somatic hybrids analysed by in situ hybridization.

In situ hybridization was used to examine genome reorganization in asymmetric somatic hybrids between Nicotiana plumbaginifolia and Nicotiana sylvestris obtained by fusion of gamma-irradiated protoplasts from one of the parents (donor) with non-irradiated protoplasts from the other (recipient). Probing with biotinylated total genomic DNA from either the donor or the recipient species unequivocally identified genetic material from both parents in 31 regenerant plants, each originating from a different nuclear hybrid colony. This method, termed genomic in situ hybridization (GISH), allowed intergenomic translocations containing chromosome segments from both species to be recognized in four regenerants. A probe homologous to the consensus sequence of the Arabidopsis thaliana telomeric repeat (5'-TTTAGGG-3')n, identified telomeres on all chromosomes, including 'mini-chromosomes' originating from the irradiated donor genome. Genomic in situ hybridization to plant chromosomes provides a rapid and reliable means of screening for recombinant genotypes in asymmetric somatic hybrids. Used in combination with other DNA probes, it also contributes to a greater understanding of the events responsible for genomic recovery and restabilization following genetic manipulation in vitro.

Cell Fusion↗
PubMed1990

Nuclear architecture in plants.

Structure within the nucleus of plants is becoming increasingly clear in both metaphase and interphase nuclei, although there are conflicting data about the relative positions of individual and pairs of chromosomes. At interphase, individual chromosomes may generally occupy discrete domains that are not intermixed with other chromosomes. Aspects of mechanical chromosome behaviour and even of gene expression may correlate with interphase chromosome position, and imply that a better understanding of nuclear architecture is required.

Cell Nucleus↗
PubMed1990

Genomic in situ hybridization to sectioned nuclei shows chromosome domains in grass hybrids.

In situ hybridization using biotinylated total genomic DNA and avidin detection systems was adapted for examination of thin-sectioned plant material in the light and electron microscopes. Root tip material was preserved prior to sectioning, so that the in vivo disposition of the chromatin was maintained. Use of total genomic DNA from Secale africanum as a probe enabled the chromatin from the two parental genomes in the grass hybrid Hordeum chilense x S. africanum to be distinguished. The biotinylated probe preferentially labelled the chromosomes of S. africanum origin. DNA-DNA hybrids were visualized at the light-microscope level by Texas Red fluorescence and at the electron-microscope level by the enzymic precipitation of DAB (diaminobenzidine) or by colloidal gold particles. The use of thin sections allowed the location of probe hybridization to be established unequivocally in both metaphase and interphase nuclei. Analysis of interphase nuclei showed that chromatin originating from the two parental genomes did not intermix but occupied distinct domains.

Chromatin↗
PubMed1989

The volumes and morphology of human chromosomes in mitotic reconstructions.

Ten unpretreated normal human male fibroblast cells in mitosis were completely reconstructed from micrographs of between 82 and 119 consecutive serial sections. All 46 chromosomes and their centromeres could be reconstructed in every cell. Measurements of chromosome volumes and centromere indices are presented. The data enabled allocation of all chromosomes to their groups (A to G), and chromosomes 1, 2, 3, 6, 9, 16, 17, 18 and Y were individually identified. Comparisons with published karyotypes showed that volume measurements correlated well with measurements of DNA content and chromosome length. Centromere indices also showed good correlation, but the acrocentric chromosomes were more unequally armed than found by length measurement. Secondary constrictions at the nucleolar organising region were visible on about a third of the acrocentric chromosomes. One chromosome of the C group, number 9, had a diffuse subcentromeric region (DSR) on the long arm, at the position of the constitutive heterochromatin and (in meiotic cells) the paramere.

Cells, Cultured↗
PubMed1989

Replicon size and rate of DNA replication fork movement are correlated in grasses.

In eukaryotes, most nuclear DNA replication proceeds bidirectionally from multiple origins of replication. A unit of DNA, replicated by two replication forks from a single origin, is termed a replicon. Using results from DNA fiber autoradiography we show a novel positive correlation between replicon size and the rate of replication fork movement in root meristem nuclei of 13 grass species. Although there is interspecific variation in replicon size, it is balanced by similar variation in the rate of replication fork progression.

Poaceae↗
PubMed1985

Community-oriented medical education: comparison of tracks in Mexico and New Mexico.

Most major innovations in medical education have occurred in a small number of new schools. Attempts to create change in traditional schools are far more complex. Relevant models for such change are few and there is an urgent need for institutions pioneering such changes to learn from each other's experiences. Two conventional medical schools described in this paper have attempted to create community-oriented institutional change by establishing experimental undergraduate curricular tracks. One is in a developing country, Mexico, the other in a technically developed country, the United States. These new tracks in medical education evolved independently of one another, yet they have many similarities and have experienced common problems. A formal exchange between the two schools has led to new insights in medical education and improvements to both programmes. Exchange agreements like the one reported here would be valuable for other medical schools in both developing and technically developed countries.

Community Medicine↗
PubMed1985

Interlocked bivalents in reconstructed metaphase I cells of bread wheat.

Complete reconstructions of all the bivalents were made from electron micrographs of serial sections through six pollen mother cells at metaphase I of meiosis in Triticum aestivum (hexaploid bread wheat). At least two of these metaphases contained interlocked pairs of bivalents. In one, two ring bivalents were interlocked, while in another a rod bivalent ran through the centre of a ring bivalent. Two other groups of bivalents were too closely appressed to allow separation into individual bivalents and may have contained interlocks. Meiosis in other anthers of the same plants examined by light microscopy was considered normal. The frequency of interlocking found was much higher than reported from light-microscope spreads. Not all interlocks in metaphase I cells need adversely affect meiosis, but knowledge of their regularity and form may facilitate understanding the processes of chromosome pairing.

Centromere↗
PubMed1984

Premeiotic events and meiotic chromosome pairing.

There is practical difficulty in identifying when meiosis begins. Moreover, because of contradictory definitions there is ambiguity and some confusion as to when, in terms of the cell cycle, premeiosis ends and meiosis begins. Nevertheless, results for several organisms show clearly that meiotic chromosome behaviour is affected by premeiotic events and especially by events during the final premeiotic mitosis and/or premeiotic interphase. This review considers only premeiotic events which do (or might) affect meiotic chromosome pairing by their effect on genomic characters, such as: chromosome number, homology, condition and position, with particular emphasis on the last. Interpreted in its widest sense 'premeiotic events affecting meiotic chromosome pairing' must include karyogamy. Moreover, while karyogamy is the normal means of achieving the diploid chromosome number and pairs of homologues essential for normal chromosome pairing, it is not the only way, as illustrated by the remarkable premeiotic adaptations seen in the apogamous ferns and the frog Rana esculenta. Little is known about the condition (including the molecular organization) of chromosomes during their approach and switch to meiosis. However, completion during premeiosis of some DNA synthesis may be essential for normal meiotic chromosome pairing. Various results (including different effects of colchicine given first at different premeiotic stages) have been claimed as evidence of one or other type of premeiotic spatial ordering of chromosomes which might favour, or be essential for, meiotic chromosome pairing. Chromosome placement has been studied recently using the electron microscope, serial thin-section, reconstruction technique. This has revealed clear evidence of non-random spatial placement of chromosomes in non-meiotic and premeiotic cells. For example, in root-tip cells of barley, Hordeum vulgare L. cv. Tuleen 346 (2n = 2x = 14), it showed: a significant spatial separation of two haploid chromosome sets; no somatic association of homologues; but a significant expression of a predicted mean order of heterologues (namely, linkage groups -4-1-2-3-5-6-7-) within haploid sets. This spatial arrangement of chromosomes might have important implications for the switch from premeiotic to meiotic development, if it occurred in germ line cells. Consequently, metaphase cells of Tuleen 346 at the final premeiotic mitosis were reconstructed next. Analysis of centromere positions showed a significantly ordered arrangement of haploid sets, and of heterologues within haploid sets, essentially the same as that previously noted for root-tip cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Ordering decision and clinic cost variation among resident physicians.

Physicians vary considerably in the services they order for their patients. We examined ordering variation among 47 resident physicians during 4,991 continuity clinic visits with patients who had specific, chronic medical problems. We ranked the physicians by their average charge per visit and grouped them into three equal categories. High-charge physicians averaged $164 per visit, medium-charge $124 and low-charge $97. In comparing the frequencies with which physicians in each group ordered a wide array of specific laboratory tests, x-ray studies, medications and miscellaneous items, we found that ordering variation among the physician groups was not confined to certain decisions or categories of services. High-charge physicians ordered a little more of nearly every item or service. Although the magnitude of ordering variation for each item was small, the sum over many items was great, suggesting that cost-containment efforts may have to focus on the "little ticket" decision-making style of expensive physicians.

Cost Control↗
PubMed1984

Chromosome order--possible implications for development.

Chromosomes are arranged in ordered haploid sets around the centre of the metaphase plate at mitosis in several grass species and hybrids. Each chromosome is in a fixed mean position relative to other, heterologous chromosomes, this order can be predicted using Bennett's model, and is clearly demonstrated from reconstructions of electron micrographs of serial sections (see Heslop-Harrison & Bennett, 1983a,b,c). The nucleus contains spatial domains of genes with similar functions. Chromosomes with major effects on nuclear behaviour--division or meiotic pairing--may be at special positions in the order. Changing spatial relationships of chromosomes with respect both to each other and the nuclear envelope (during the cell cycle and during development) may affect cell differentiation and gene activity. Chromosome order may have implications for the control of development within the nucleus and the organism. Order may constrain karyotype and hence species evolution.

Cell Differentiation↗
PubMed1983

Impact of a cost-containment educational program on housestaff ambulatory clinic charges.

A cost-containment project was established to determine if a broad educational program that coupled clinical logic with feedback of charges to internal medicine residents in an ambulatory clinic setting could alter behavior. The project was evaluated by comparing charge data from a preintervention comparison year with charge data from the intervention year. Decreases in charges for laboratory procedures occurred in six of the seven diagnostic categories during intervention and were significant (t-test) for four categories. Regression analysis evaluated the influence of the intervention and 12 other variables on charges and demonstrated that the intervention was associated with a mean reduction of lab charges of $ 6.30 (p less than 0.0001) and a mean reduction of total encounter charges of $ 10.36 (p less than 0.01). We conclude that our educational program resulted in a decrease in mean laboratory changes and total charges for ambulatory encounters.

Awareness↗
PubMed1983

Comparison of family medicine and internal medicine: charges for continuing ambulatory care.

The authors compared charges for ambulatory and continuing patient care prescribed by residents in internal medicine and family medicine. An analysis of covariance showed that the charges per encounter in internal medicine were $38.83 greater than charges in family medicine after accounting for differences such as patient age, diagnosis, and severity of condition. However, family practitioners scheduled patient encounters more frequently. The median number of days between encounters was 30 for family medicine and 60 for internal medicine. The duration of medical care for each patient (up to 18 months was possible) and the summation of encounter charges over that duration of medical care were evaluated. Because only two out of eight duration-of-medical-care categories were significantly less in family medicine, the differences in charges between the specialties essentially disappeared. Comparative studies which consider only charges per encounter potentially have a large bias.

Analysis of Variance↗
PubMed1983

DNA density in mitotic and meiotic metaphase chromosomes of plants and animals.

Studies of chromosome disposition at metaphase using serial thin-sectioning and three-dimensional reconstruction techniques have produced accurate estimates of the total volume of chromosomes per cell in 15 plant and two animal species. Comparing this character with the 4C DNA amount showed no indiction of systemic differences in DNA density between either organisms with widely different (greater than 200-fold) C values or different groups or organisms. For example, there was no significant difference between the density of DNA in somatic metaphase chromosomes of man (0.141 pg/micrometers3) and its mean in 14 angiosperm plant species (0.182 pg/micrometers3), or between four dicotyledons (0.180 pg/miocrometers3) and 10 monocotyledons (0.182 pg/micrometers3). However, evidence was found showing that DNA density can vary significantly within a species. Thus, although the total chromosome volume per cell was closely correlated (r greater than 0.97) with 4C DNA amount in somatic and meiotic cells, the density of DNA in metaphase chromosomes was significantly lower in meiocytes (0.131 pg/micrometers3) than in somatic metaphase cells (0.19 pg/micrometers3).

Animals↗
PubMed1983

The positions of centromeres on the somatic metaphase plate of grasses.

Centromere positions on the metaphase plate of 48 root-tip cells of four grass species were analysed using metaphases reconstructed from electron micrographs of serial thin sections. Centromere alignment was almost perfect on a plane in 15 untreated metaphases of cereals where 2n = 14; only 2% of the total variance in centromere position was out of the plane of the metaphase plate. In 23 similar cells pretreated with ice-water, the mean out-of-plate variance was 9%, compared to an expectation of 18% if centromeres were positioned randomly in space. In cold-treated cells of Zea mays (2n = 20), the out-of-plate variance (14%) was significantly less than a random expectation of 20%. The distances of centromeres from the mean centromere position (MCP) were also analysed. They showed that the centromeres tended to be normally distributed about a circle around the MCP in all the species. Thus centromeres tend towards a ring arrangement in metaphase cells. Analysis of separation distances of all pairs of centromeres in each reconstructed cell showed that the distribution of distances between centromeres is not random: there are significantly fewer centromeres that are close together and more that are at medium separation distances than is expected on a random basis. This is different from previously published assumptions about centromere disposition in squashed metaphases.

Centromere↗