Bone marrow changes after localized acute and fractionated X irradiation.
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Biomedical subjects
Publications and source records attributed to A L Carsten.
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These studies were to evaluate the effects of humoral factors on amplification of nonrecognizable erythrocytic and granulocytic precursors by the in vivo plasma clot diffusion chamber and the in vitro plasma clot culture methods. Changes in the plasma erythropoietin levels in the reticulocyte concentration and hematocrits of irradiated and nonirradiated Long-Evans rats exposed to hypoxia were also determined. While erythropoietin plasma concentrations appeared to affect BFU-E and CFU-E growth, results suggest erythropoietin may not be the sole regulator of red cell production and that inhibitors of chalone-like mechanisms may be involved. Measurements made on granulocyte precursors treated with colony stimulating factor (CSF) containing L-cell-conditioned medium revealed granulocytic colonies and burst-like formations similar to those seen for erythrocytic growth. There is strong evidence suggesting that CSF is a regulator of granulopoiesis; however, it is not the sole regulator and it appears that inhibitors may play an in vivo role. Growth of colonies with cell numbers not a power of 2 implies either asymmetric mitosis due to loss of genetic information required for continuing division, or differences in concentrations of, or ability to recognize, inhibitory factors. These possibilities are examined in the light of results from in vivo and in vitro culture techniques.
Nucleated cells obtained from blood and/or bone marrow of patients with myelofibrosis with myeloid metaplasia (MMM) were cultured in diffusion chambers (DC) implanted into the peritoneal cavities of irradiated mice. A total of five blood studies and two bone marrow studies were performed using cells obtained from five patients. The DC were harvested at intervals from the host mice and the total and differential cellularity of DC contents were evaluated. The results obtained from MMM cultures were compared with those from similar cultures of blood cells and marrow cells of four and six normal individuals respectively. The proliferation and maturation of the granulocytic, erythrocytic, and megakaryocytic lines in MMM cultures occurred in an orderly fashion as they occur in vivo. The patterns of proliferation and maturation of the three cell lines in cultures after day 7 suggest that they primarily originate from progenitor cells. The numbers of granulocytes in the multiplicative pool, recognizable red cell precursors, and megakaryocytes recovered were significantly greater from the MMM cultures than those from the normal blood or marrow cultures. These results suggest that the blood and marrow cells of MMM patients have increased numbers of progenitors for granulocytes, erythrocytes, and megakaryocytes.
The effect of X-rays and different-energy neutrons on human bone-marrow cells was studied using two different cell-culture techniques--diffusion chamber (DC) growth and colony formation in vitro (CFU-C). Based on the survival of proliferative granulocytes in DC on day 13, the D0 value was 80 rad with X-rays, and 117 rad as measured by the CFU-C assay. The D0 values for neutrons depended on the radiation source and the energy level. The r.b.e. values, which dropped with increasing energy levels of mono-energetic neutrons, were (i) 0.44 MeV; DC 3.7, CFU-C 4.1; (ii) 6 MeV; DC 1.8, CFU-C 2.0; (iii) 15 MeV; DC 1.6, CFU-C 1.6; (iv) fission neutrons; DC 2.6, CFU-C 2.4.
Radiation damage to the right femoral artery was studied in mice 7--8 weeks old. Single X-ray exposures of 500 to 10 000 rad were used. The animals, except for the right upper legs, were shielded from exposure. The unexposed left femoral arteries served as 'controls'. Animals were killed at post-irradiation times of up to 120 days. Pathological changes in the irradiated arteries were scored on an arbitrary scale of 0 to 5 for individual tunica. The overall arterial damage was scored on a scale of 0 to 15. Radiation doses of 1000 to 10 000 rad produced quantitative histological changes in the three individual tunica and the degree of damage was dose-dependent. The tunica adventitia showed the maximum radiation injury, followed by tunica media and tunica intima in decreasing magnitude of injury. The combined effects on the artery followed closely the effects on media and adventitia.
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A transplanted tumor that induces granulocytosis and produces colony stimulating factor (CSF) was studied in mice during several passages. The sequence of events leading to granulocytosis was characterized. Band granulocytes were increased 3 days after tumor inoculation, while simultaneously CFU-s and CFU-c in bone marrow and spleen were transitorily low. This was followed by rapid accumulation of CFU-c and CFU-s in spleen, and by successive waves of increased mitotable and non-mitotable granulocytes in spleen and marrow. In contrast, marrow CFU-c and CFU-s remained normal or slightly decreased.
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The long term somatic and genetic effects of chronic tritium (3H) ingestion have been measured in mice. Second generation animals, who with their parents were maintained on drinking water containing 3 muCi/ml of HTO, have been bred and the number of dominant lethal mutations measured. The increase in dominant lethal mutations in treated animals as compared to tap water controls is significant at the 1% level. The stem cell content of the bone marrow has been measured throughout the lifetime of the animals. A slight reduction in colony forming units in the bone marrow of treated animals is evident beginning after approximately 12 weeks, increasing in severity throughout the lifetime of the animals. The distribution of tritium in tissue water protein fractions was determined, and the relative contribution to tissue dose calculated. The differential incorporation into protein fractions was not found to be significant.
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The spleen-colony technique has been used for determining the relative biological effectiveness (r.b.e.) for several energies of neutron radiation. Donor mice were exposed for fission and accelerator-generated neutrons at a variety of doses and energies. Immediately after exposure, donor bone-marrow was removed from the hind legs, and standard amounts were injected intravenously into lethally X-irradiated recipients. After 7 days the recipients spleens were evaluated for surface colonies. Dose-response curves were obtained for each type of radiation and the Do was determined. The neutron r.b.e. values from the Do compared with 250kVp X-rays were: reactor 1.58, 252Cf 1:59, and accelerator varied from 2.85 at 1.0 Mev to 0.85 at 13.4 MeV.
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In order to investigate some characteristics of Millipore diffusion chamber cultures (DCC) and their potential for the quantitation of tumor cell growth kinetics, HeLa S-3-cells were grown in diffusion chambers implanted i.p. in mice. HeLa cells grew pseudologarithmically in DCC with a population-doubling time of approximately 2.8 days when 9.2 to 12.0 X 10(5) cells were placed in the chambers initially. The population-doubling time varied as a function of the initial innocula size but was always longer than the in vitro population-doubling time of 1.2 days. Comparable harvests of cells from experiment to experiment varied quantitatively by no more than a factor 1.8. This variaton somewhat limits the use of DCC for the quantitation of subtle effects. The fate and morphology of HeLa cells in DCC were determined from scanning electron microscopy. DCC might be exploited in clinical studies of the growth of patients' tumors and the effects of various therapies in a host-mediated system.
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A survey of the literature to date on the enzyme histochemistry of intracellular organelles has not yielded any reference to the presence of acid phosphatase reaction products in the mammalian mitochondria of the central nervous system. A combination of Gomori's acid phosphatase mehtod, however, with standard electron microscopy has disclosed the presence of enzyme reaction products in the mitochondria of the central nervous system of rats from 2 hr to 22 weeks after x-ray irradiation, as well as in a cerebral biopsy performed on a patient affected by Huntington's chorea. No enzyme reaction products, on the other hand, were observed in serial sections that had been incubated in substrates either containing sodium fluoride or lacking in beta-glycerophosphate. The abnormal mitochondrial enzyme reaction (chemical lesion) is considered to be the consequenco of the pathologic process affecting the ultrastructural-chemical organization of the organelle.