Biomedical subjects

K M Townsend

Publications and source records attributed to K M Townsend.

At least 37 records · Page 2Linked to original sources

PubMed1996

Differentiation and delayed cell death in embryonal stem cells exposed to low doses of ionising radiation.

Embryonal stem cells have been used to study the effects of environmentally relevant doses of radiation on cell death and differentation. The ES cells were found to have a greater than 60% chance of surviving the traversal of a single alpha-particle, the lowest possible dose of high linear energy transfer radiation a cell may receive. The ES cells appeared to possess the cell cycle checkpoints believed to prevent the transmission of the radiation damage. However, delayed effects were observed in the progeny. An increased incidence of apoptosis and haempoietic differentiation capacity was found to persist in the ES cell population over many cell divisions. Since both cell death and differentiation are known to play a key role in tissue kinetics, an ES cell model will provide a valuable and versatile cell system for studying the role of cell death and differentiation in the pathology of radiogenic diseases.

Journal Article↗

PubMed1995

Radiation-induced genomic instability: delayed cytogenetic aberrations and apoptosis in primary human bone marrow cells.

Transmissible chromosomal instability, characterized by non-clonal cytogenetic aberrations with a high frequency of chromatid-type aberrations together with a lower frequency of chromosome-type aberrations, has been demonstrated in the clonal descendants of human haemopoietic stem cells after alpha- but not X-irradiation. Comparable cytogenetic abnormalities have also been demonstrated in non-clonal cultures of alpha-irradiated primary human bone marrow, but a different pattern of delayed aberrations, mainly of chromosome-type, was found after X-irradiation in non-clonal cultures. In clonal analyses, delayed apoptotic cell death was evident after both X- and alpha-irradiation. It is suggested that the type of radiation exposure, the type of cell and its genetically determined susceptibility are factors that may influence the expression of delayed effects of radiation.

Alpha Particles↗

PubMed1995

The effect of dimethyl sulfoxide on the induction of DNA double-strand breaks in V79-4 mammalian cells by alpha particles.

The present study was undertaken to assess the protective effect of dimethyl sulfoxide (DMSO) against the induction and rejoining of DNA double-strand breaks (DSBs) and inactivation of V79-4 Chinese hamster cells by both high- and low-linear energy transfer (LET) radiations. The cells were exposed under aerobic conditions as monolayers to either low-LET photons (60Co gamma rays) or high-LET alpha particles (238Pu) at 277 K. The initial yield of DSBs, determined by elution under nondenaturing conditions, is linearly dependent on dose. When the irradiation was carried out in the presence of DMSO (0-0.6 mol dm-3), the initial yields of DSBs induced by both gamma and alpha-particle irradiation decrease. With gamma irradiation at [DMSO] > 0.6 mol dm-3, a further decrease in the yield of DSBs occurs. DMSO (0.5 mol dm-3) reduces the initial yield of DSBs by 50 +/- 5% and 32 +/- 4% for photons and alpha particles, respectively. DMSO protects more effectively against cellular inactivation and DSB induction at low LET compared with alpha-particle irradiation with protection factors of 1.7 and 1.4, respectively, for survival and 2.0 and 1.5, respectively, for DSBs. After incubation of the irradiated cells for 3 h at 310 K after high-LET irradiation, the residual yield of DSBs is reduced by < 13% when the irradiations were carried out in the presence of 0.5 mol dm-3 DMSO. With gamma irradiation in the presence of 0.5 mol dm-3 DMSO, 90% of the DSBs are rejoined by 3 h incubation at 310 K. Therefore, the nonscavengeable DSBs induced by alpha particles are not significantly rejoined within 3 h, in contrast to rejoining of the majority of the nonscavengeable DSBs induced by gamma irradiation. From comparison of the data on DSBs and survival for alpha-particle irradiation, it is inferred that the severity of damage is reduced by DMSO through minimizing the formation of OH-induced sugar/base modifications in the vicinity of nonscavengeable DSBs.

Alpha Particles↗

PubMed1993

Field alternation gel electrophoresis--status quo.

Since the description of the original technique of field alternation gel electrophoresis (FAGE) about ten years ago there have been significant developments in the area. Between 1983 and early 1987 dramatic improvements in the technique and apparatus resulted in a 500- to 600-fold increase in the functional separation capacity of conventional agarose gel electrophoresis. Details of the improvements in technique and equipment was the subject of an earlier review [H. J. S. Dawkins, J. Chromatogr., 492 (1989) 615]. This review concentrates on the application of FAGE technology. The FAGE technique is no longer restricted to simply separating large DNA fragments. This method is presently being used for electrophoretic karyotyping, long-range genomic mapping, cloning of large DNA fragments into new vectors, the study of pathogenic chromosomal alterations and the structural analysis of chromosomes. The applications of FAGE in molecular biology and genetics is constantly expanding, with the full potential of this technique still to be realised.

Animals↗

PubMed1992

Nuclear area measurement on viable cells, using confocal microscopy.

We describe a rapid procedure for the accurate measurements of nuclear areas on unperturbed living cells as used in radiobiological experiments, using the confocal laser scanning microscope. The microdosimetric interpretation of radiobiological data requires precise information on the nuclear area of cells as irradiated with high-LET radiation.

Animals↗

PubMed1992

The influence of type I collagen on the growth and differentiation of the human colonic adenocarcinoma cell line HT-29 in vitro.

HT-29 Human colonic adenocarcinoma cells when grown on a plastic substratum were anaplastic in appearance and failed to express any morphological or biochemical features that were characteristic of intestinal differentiation. Growth of HT-29 cells subcutaneously in the flank of immune deprived mice gave rise to morphologically heterogeneous tumors which were poorly differentiated but contained approximately 11% of cells with an intestinal phenotype: these showed features typical of cell polarization with well-developed microvilli, tight junctional complexes and desmosomes between adjacent cells. The transfer of cells from plastic onto either a fixed (designated 'non-released') or floating (designated 'released') type I collagen gel induced some morphological features typical of intestinal differentiation; for example goblet-like cells were observed after 9 days, but biochemical markers of differentiation were expressed only modestly. The continued subculture of HT-29 cells on collagen type I gels, which were either attached to the plastic or floating in the medium, induced some morphological features of intestinal differentiation and changes in the activity of brush border-associated enzymes. Alkaline phosphatase activity was enhanced from 1.3 x 10(-3) mumoles/mg/min for cells cultured on plastic substrata to 2.1 x 10(-3) mumoles/mg/min when gels were non-released, and 2.9 x 10(-3) mumoles/mg/min when gels were released after 12 days of culture. This was confirmed by electron microscopical visualization of alkaline phosphatase activity. Elevated levels of aminopeptidase activity were also observed on day 12 (plastic = 26 milliunits/mg; non-released gel = 41 milliunits/mg; released gel = 36 milliunits/mg). Similarly, changes occurred in the secretion of carcinoembryonic antigen from 0.96 x 10(-2) micrograms/mg/48 hours by cells cultured on plastic to 2.3 x 10(-2) micrograms/mg/48 hours by cells cultured on floating collagen gels. The effects of permitting HT-29 cells to undergo polarization were tested by culture on inert filter inserts: morphological features of intestinal differentiation were observed although this did not occur until after 21 days. These studies show that optimization of the growth conditions of anaplastic cells in vitro may provide cultures more representative of the tumor in vivo. This model system may be useful for cell biological and pharmacological studies of colon carcinoma.

Adenocarcinoma↗

PubMed1991

Role of the extracellular matrix on the growth and differentiated phenotype of murine colonic adenocarcinoma cells in vitro.

The growth and differentiation characteristics of MAC 15 murine adenocarcinoma cells, derived from routine passage in vivo for growth in vitro on a plastic substrate (MAC15j cells), were compared under conditions in which the cells were seeded onto a substrate of type-I collagen which was either attached to plastic or was released to float free in medium. Cells grown on a plastic substrate consisted of a heterogeneous, largely anaplastic population with a putative enterocytic morphology but with no evidence of junctional complexes or cell polarity typical of an epithelial phenotype. MAC 15j cells from cultures grown on a plastic substrate reestablished a moderate to well-defined degree of differentiation when transplanted back into NMRI mice. When MAC 15j cells were seeded from plastic onto type-I collagen, either attached to plastic or free-floating, tight junctional complexes were formed and the cells began to attain a more recognizable, columnar and polarised epithelial morphology. Cells grown on a type-I collagen gel which was free-floating showed a selective expression of alkaline phosphatase at the apical surfaces of approximately 10% of the cells. This expression was detectable by electron microscope histochemistry but could not be detected biochemically. Treatment of MAC 15j cells grown on a released collagen matrix with tetramethyl-urea (20mM) accelerated the expression of alkaline phosphatase activity at the apical surface as detected by microscopy.

Adenocarcinoma↗

PubMed1990

Thickness measurements on V79-4 cells: a comparison between laser scanning confocal microscopy and electron microscopy.

A quantitative comparison has been carried out between laser scanning confocal microscopy on living cells and standard electron microscope methods on fixed samples. It was estimated from these measurements that there was about 10-20% reduction in thickness in fixed samples of monolayer V79-4 hamster cells. Precise information on the true thickness of living cells, as irradiated, is required for full interpretation of radiobiological data with poorly penetrating radiations, including ultrasoft X-rays. The confocal microscope allows rapid measurements on unperturbed living samples.

Animals↗

PubMed1989

Studies of alveolar cell morphometry and mass clearance in the rat lung following inhalation of an enriched uranium dioxide aerosol.

Seventy-three rats were exposed to an aerosol of enriched uranium dioxide (UO2), giving initial lung burdens of 26 to 447 micrograms at 6 days post-inhalation (PI). At 7 days PI 35 of these rats were further exposed to thermalised neutrons at a fluence of 1 x 10(12) neutrons cm-2. There was no significant difference between the two groups in the clearance rate of the UO2 particles from the lung, up to 590 days PI. The particles cleared relatively slowly over this period with a retention half-time in the lung of 160 to 176 days. Transmission electron microscope (TEM) studies of tissue from the alveolar region at 8 days PI showed that inhalation of UO2 particles significantly increased the sizes of macrophage and type II cells, and the number of macrophage and type I cells. There was also a significant increase in the size of lysosomal granules within the macrophages after exposure to the UO2 particles. The exposure to UO2, neutrons and 235U fission fragments had no significant effect on any of the cells above that observed in the animals exposed to UO2 alone. Additional rats were exposed to the same neutron fluence without prior UO2 inhalation. The alveolar cells of neutron-only exposed rats were, in size and number, typically no different from those in the completely unexposed control rats.

Administration, Inhalation↗

PubMed1987

Quantitative studies of the fate of epidermal Langerhans cells after X-irradiation of guinea-pig and mouse footpad skin.

Langerhans cell numbers, morphology and distribution were observed in cross sections of footpad epidermis at intervals from 1 to 28 days after exposure of the hind feet of CBA/H mice or albino guinea-pigs to a single absorbed dose of 20 Gy (2000 rad) of X-rays. In mice, the number of Langerhans cells reactive with anti-macrophage F4/80 monoclonal antibody steadily declined by approximately 85% within 10 days after irradiation, consistent with previous studies, in which Langerhans cells were identified in epidermal sheets by ATPase activity or presence of Birbeck granules. Remaining Langerhans cells were exceptionally dendritic. Very few Birbeck granule-containing cells were found in murine popliteal lymph nodes before or after irradiation but damaged cells were present in superficial strata of irradiated epidermis. The morphology and number of epidermal F4/80-positive cells approached normal by 15 days after irradiation. In guinea-pigs, gradual suprabasal movement and loss of rounded, ATPase-positive Langerhans cells from the epidermis were detectable from 5 to 20 days after irradiation but the magnitude of the cell loss and redistribution was partially obscured by the simultaneous appearance of clusters of replacement Langerhans cells in the basal layer and by keratinocyte hyperplasia.

Adenosine Triphosphatases↗

PubMed1987

Subcutaneous implantation of double velour Dacron into the mouse: infiltration and angiogenesis.

An experimental system has been devised for the study of tissue reaction to the subcutaneous implantation of double velour Dacron into the mouse. Animals were given Dacron implants for 3 months, 2 months, 1 month, 3 weeks, 2 weeks and 1 week and the infiltration of the material was assessed using light-microscopy, autoradiography, electron-microscopy and angiography. It was found that the implants became extensively infiltrated with host cells, the response being at a peak in the second and third weeks post-implantation. Macrophages were seen from an early stage, fibroblasts were numerous, and new capillaries penetrated the material. The observations, especially the angiogenic response, are discussed with reference to published information on the actions of the cell types that were seen, in particular the macrophage.

Angiography↗

PubMed1985

Man's sensitivity to bone marrow failure following whole body exposure to low LET ionising radiation: inferences to be drawn from animal experiments.

Evidence concerning the sensitivity of man to bone marrow failure following exposure to brief but substantial doses of ionising radiation is sparse. There is, however, a relatively substantial body of information on such effects in large animals. Reported experiments on six species where exposure to low LET radiation was uniform to the whole body and of brief duration (exposure times of the order of one hour or less) have been reanalysed both in terms of exposure and of midline tissue dose. The results indicate a marked lack of homogeneity among values for LD50 within species thus questioning the applicability of LD50 as a species dependent constant. It is, however, suggested that on a purely empirical basis these large animal data suggest that the dose killing 'most' (where 'most' is between 90 and 95 per cent) is about twice that killing 'few' (where 'few' is between 5 and 10 per cent). For man, where there is evidence that the dose killing few is unlikely to be less than 3 Gy, this relationship might indicate a gradient of mortality with dose between 3 and 6 Gy.

Animals↗

PubMed1985

Dose response of Langerhans cells in mouse footpad epidermis after X irradiation.

Effects of a range (2-50 Gy) of single doses of 250 kV X rays on epidermal Langerhans cells in vivo were quantified in groups of CBA/CaH mice. Animals were sacrificed and compared with controls on the 10th day after local irradiation of their hind feet, when Langerhans cell numbers were at a minimum. ATPase-positive Langerhans cells in sheets of footpad epidermis were counted by light microscopy and cells with Birbeck granules were enumerated by electron microscopy. Both methods revealed a dose-dependent loss of Langerhans cells after ionizing radiation. Fifty percent of the ATPase-positive cells were lost after 14.4 +/- 1.3 Gy, and 50% of Birbeck granule-containing cells were lost after 17.9 +/- 4.2 Gy, suggesting that differentiated epidermal Langerhans cells are radioresistant. Loss of equivalent proportions of ATPase-positive and ultrastructurally identifiable cells after a range of doses indicates that X rays do not merely alter Langerhans cell surface markers but actually deplete the epidermal population of these cells.

Animals↗

PubMed1984

Langerhans cell number and morphology in mouse footpad epidermis after X irradiation.

Effects of 250-kV X rays on epidermal Langerhans cells were studied in CBA/CaH mice. One group received 20 Gy to the feet, another 8 Gy to the whole body, and a third both the 8 Gy whole-body and a 12 Gy local dose to the feet. Mice from each group and controls were sacrificed at intervals from 1 to 64 days later. ATPase-positive cells in sheets of footpad epidermis were counted by light microscopy. The density of Langerhans cells in controls was 1515 +/- 36/mm2 (mean +/- SE; n = 34). By 3 days after irradiation they became rounded and less dendritic and numbers gradually reached a nadir by 10 days, at 18% of controls after 20 Gy and 57% of controls after 8 Gy. Some of the remainder exhibited bizarre morphology and ultrastructural abnormalities. After local irradiation of the feet Langerhans cell numbers recovered rapidly between 14 and 16 days, although their distribution was uneven until 30 days after irradiation. Repopulation was delayed after an 8 Gy whole-body dose by at least 3 weeks. These results demonstrate that high local doses of X rays substantially but transiently deplete the epidermal Langerhans cell population and support the hypothesis that functional hemopoietic tissue is required for extensive Langerhans cell replenishment.

Animals↗

PubMed1984

The effects of X irradiation on the cytoskeleton of rat alveolar macrophages in vitro.

The three-dimensional visualization of Triton X-100 resistant cytoskeletons has been used to demonstrate that an absorbed dose of 120 Gy from X rays causes a distinctive and reproducible alteration of the cytoskeleton of intact rat alveolar macrophages in vitro. The alteration has also been shown to be rapidly and completely "repaired" and to be apparently similar to alterations caused by colchicine but dissimilar to those caused by cytochalasin B. From these observations and those of other workers who have studied the irradiation of extracted microtubular proteins in vitro, we think it likely that microtubules rather than microfilaments are the radiosensitive component of the macrophage cytoskeleton.

Animals↗

PubMed1982

Longevity of osteoclasts in radiation chimaeras of beige and osteopetrotic microphthalmic mice.

Beige (bg/bg) mice lethally X-irradiated (900 or 1000 rad) and rescued with spleen cells from osteopetrotic (mi/mi) mice were examined periodically up to 6 months thereafter for signs of osteopetrosis, radiographic and histological. Their metaphyses were also examined electromicrographically for general characteristics of osteoclasts and for giant lysosomal granules. Measurements of lysosomal area indicated no change in the first weeks, then an exponential decline to near base line after 3 months with a half-period of or approximately 10 days. In this combination osteopetrosis was of slower progression than hitherto reported in C57BL mice. We suggest that osteoclasts have no fixed life span and that their longevity is a function of genetic background and perhaps operational needs.

Animals↗

PubMed1982

Longevity of osteoclasts in radiation chimaeras of osteopetrotic beige and normal mice.

Beige mice, osteopetrotic from incorporated mi/mi genes, and their skeletally normal siblings were X-irradiated and converted to radiation chimaeras by rescue with normal CBA bone marrow. The procedure produced rapid cure of the osteopetrosis. Electron micrographic measurement of the decline in giant lysosomes of the host mice indicated that the longevity of the host's osteoclasts was up to 30 days in the osteopetrotic and 40 days in the skeletally normal mice. In each case the decline was roughly exponential with half-times of about 6 and 8 days respectively. Monogenetic and perhaps polygenetic and teleological factors are invoked. Absence of a latent period before decline in the score seen in the companion paper is attributed to the presence of partially mature osteoclast precursors in bone marrow, not present in spleen.

Animals↗

PubMed1980

Osteoclasts derived from haematopoietic stem cells.

The origins of the multinucleated osteoclast have been controversial, with osteogenic precursors and haematopoietic stem cells as candidates. Recent evidence for the latter is persuasive but circumstantial. We report here direct evidence obtained in radiation chimaeras from a natural cytoplasmic cell marker transmitted by the donated haematopoietic stem cell.

Animals↗