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Biomedical subjects

P Todd

Publications and source records attributed to P Todd.

At least 109 records · Page 6Linked to original sources
PubMed1984

Electrophoretic separation of kidney and pituitary cells on STS-8.

A Continuous Flow Electrophoresis System (CFES) was used on Space Shuttle flight STS-8 to separate specific secretory cells from suspensions of cultured primary human embryonic kidney cells and rat pituitary cells. The objectives were to isolate the subfractions of kidney cells that produce the largest amounts of urokinase (plasminogen activator), and to isolate the subfractions of rat pituitary cells that secrete growth hormone, prolactin, and other hormones. Kidney cells were separated into more than 32 fractions in each of two electrophoretic runs. Electrophoretic mobility distributions in flight experiments were spread more than the ground controls. Multiple assay methods confirmed that all cultured kidney cell fractions produced some urokinase, and five to six fractions produced significantly more urokinase than the other fractions. Several fractions also produced tissue plasminogen activator. The pituitary cells were separated into 48 fractions in each of the two electrophoretic runs, and the amounts of growth hormone (GH) and prolactin (PRL) released into the medium for each cell fraction were determined. Cell fractions were grouped into eight mobility classes and immunocytochemically assayed for the presence of GH, PRL, ACTH, LH, TSH, and FSH. The patterns of hormone distribution indicate that the specialized cells producing GH and PRL are isolatable due to the differences in electrophoretic mobilities.

Animals↗
PubMed1984

Properties of electrophoretic fractions of human embryonic kidney cells separated on Space Shuttle flight STS-8.

Suspensions of cultured primary human embryonic kidney cells were subjected to continuous flow electrophoresis on Space Shuttle flight STS-8. The objectives of the experiments were to obtain electrophoretically separated fractions of the original cell populations and to test these fractions for the amount and kind of urokinase (a kidney plasminogen activator that is used medically for digesting blood clots), the morphologies of cells in the individual fractions, and their cellular electrophoretic mobilities after separation and subsequent proliferation. Individual fractions were successfully cultured after return from orbit, and they were found to differ substantially from one another and from the starting sample with respect to all of these properties.

Cell Line↗
PubMed1984

Survival of cultured human cells exposed to californium 252Cf radiation at low dose rates.

Human T-1 cells were used to determine the rbe of low dose rate 252Cf (n + gamma) radiation, compared to low dose rate gamma radiation. The T-1 cells were irradiated hypoxically at room temperature, and the dose rates were as follows: a) 252Cf (n + gamma), 0.045, 0.090, and 0.270 Gy/hour; and b) 137Cs gamma radiation, 0.117, 0.246, and 0.765 Gy/hour. The RBE was obtained as the ratio of the initial slope of the 137Cs survival curves (approximately the same for all three dose rates), and the initial slope of the 252Cf survival curves (same for all three dose rates). The RBE was 5.0 +/- 1.0 for all components of the 252Cf radiation and 7.1 +/- 1.7 for the neutron components. As the dose increased, an inverse dose rate effect (more sensitive at lower dose rates) for the 137Cs survival curves was observed.

Californium↗
PubMed1984

Kinetic differences between fed and starved Chinese hamster ovary cells.

When Chinese hamster (CHO-K1) cells are grown as monolayer cultures, they eventually reach a population-density plateau after which no net increase in cell numbers occurs. The kinetics of aged cells in nutritionally deprived (starved) or density-inhibited (fed) late plateau-phase cultures were studied by four methods: (i) Reproductive integrity and cell viability were monitored daily by clonogenic-cell assay and erythrosin-b dye-exclusion techniques. (ii) Mitotic frequencies of cells from 18 day old cultures were determined during regrowth by analysing time-lapse video microscope records of dividing cells. (iii) Tritiated-thymidine ([3H]TdR) autoradiography was used to determine the fractions of DNA-synthesizing cells in cultures entering plateau phase and during regrowth after harvest. (iv) The rate of labelled nucleoside uptake and incorporation into DNA was measured using liquid scintillation or sodium iodide crystal counters after labelling with [3H]TdR or [125I]UdR. Non-cycling cells in starved cultures accumulate primarily as G1 phase cells. Most cells not in G1 phase had stopped in G2 phase. Very few cells (less than 2%) were found in S phase. In contrast, about half of the cells in periodically fed cultures were found to be in DNA-synthetic phase, and the percentage of these S phase cells fluctuated in a manner reflecting the frequency of medium replacement. Populations of both types of plateau-phase cultures demonstrate extremely coherent cyclic patterns of DNA synthesis upon harvest and reculturing. They retain this high degree of synchrony for more than three generations after the resumption of growth. From these data it is concluded that nutritionally deprived (starved) late plateau-phase cells generally stop in either G1 or G2 phase, whereas periodically fed late plateau-phase cultures contain a very large fraction of cycling cells. Populations of cells from these two types of non-expanding cultures are kinetically dissimilar, and should not be expected to respond to extracellular stimuli in the same manner.

Animals↗
PubMed1983

An evaluation of Econazole, an antifungal agent, for use in quantitative cell culture experiments.

Econazole, an imidazole derivative with high antifungal activity, was tested for cytotoxicity using five sensitive tests of human cell function in vitro: colony forming efficiency, growth rate, cell cycle distribution of a permanent epithelioid cell line (T-1E), and attachment efficiency and urokinase production by early passage human embryonic kidney cells. None of these endpoints was detectably affected by 1 microgram/ml Econazole in serum containing medium. At 3 micrograms/ml, Econazole reduced slightly the growth rate of T-1E cells but did not detectably affect the other end points.

Cell Adhesion↗
PubMed1983

A study of cell electrophoresis as a means of purifying growth hormone secreting cells.

Growth hormone secreting cells of the rat anterior pituitary are heavily laden with granules of growth hormone and can be partially purified on the basis of their resulting high density. Two methods of preparative cell electrophoresis were investigated as methods of enhancing the purification of growth hormone producing cells: density gradient electrophoresis and continuous flow electrophoresis. Both methods provided a two- to four-fold enrichment in growth hormone production per cell relative to that achieved by previous methods. Measurements of electrophoretic mobilities by two analytical methods, microscopic electrophoresis and laser-tracking electrophoresis, revealed very little distinction between unpurified anterior pituitary cell suspensions and somatotroph-enriched cell suspensions. Predictions calculated on the basis of analytical electrophoretic data are consistent with the hypothesis that sedimentation plays a significant role in both types of preparative electrophoresis and the electrophoretic mobility of the growth hormone secreting subpopulation of cells remains unknown.

Animals↗
PubMed1983

Unique biological aspects of radiation hazards--an overview.

Low orbit, geostationary, and deep-space flights differ from one another with respect to particle radiation environment, participating population size, mission duration, and biological risks other than radiation. It is proposed that all of these factors be considered in the setting of safety standards and, in particular, that the rem-dose concept is applicable only to radiations having low and intermediate linear energy transfer (electrons, protons, and helium ions), whereas the incidence of microlesions is a more meaningful indicator of the hazard due to higher-Z, high energy (HZE) particles. A microlesion is the biological injury inflicted in a specific tissue by a single HZE particle, and it is still in need of quantitative biological definition for specific mammalian tissues. If for example, a microlesion is taken as due to a HZE particle track 10 cell diameters long with LET > 200 KeV/micrometer in its core and > 25 rad dose in its penumbra at a distance of 10 micrometers, then the microlesion dose rate in geostationary orbit, for example, is about 9,000 microlesions per cm3 of tissue per month.

Aerospace Medicine↗
PubMed1979

Gamma rays: further evidence for lack of a threshold dose for lethality to human cells.

In experiments designed to measure human cell survival sith +/- 2 percent accuracy it was found that low doses (21 to 87 rad) of gamma-rays inactivated the colony-forming ability of cultured human cells with a probability of 0.00226 +/- 0.00012 per rad. There appears to be no threshold for the lethality of radiation to human cells in vitro.

Cell Division↗
PubMed1979

Rabbit tubular basement membrane. Isolation and analysis of polypeptides.

Renal tubules from rabbit kidneys were isolated from thin shavings of the kidney surface. Basement membrane was then prepared following sonication of the isolated tubules. To insure preservation of the integrity of the basement membrane polypeptides, the protease inhibitors, diisopropyl fluorophosphate, ethylenediaminetetraacetic acid, N-ethylmaleimide, and epsilon-amino-caproic acid were used at all stages of the preparations. The optimal conditions of sonication and centrifugation were established and the chemical composition of basement membrane prepared under these conditions was examined in detail. Glycine, hydroxyproline, and hydroxylysine were found in concentrations of 206, 65, and 18 residues per thousand, respectively, in basement membrane from young kidneys. About 38% of the basement membrane was found to be soluble in sodium dodecyl sulfate upon incubation at 90 degrees C, and to possess relatively low amounts of the amino acids characteristic of collagen. Electrophoretic analysis of this fraction revealed that the major subunits ranged in approximate molecular weight from 18,500 to greater than 10(6). When analyzed with disulfide bonds reduced, a molecular weight range from 31,000 to 275,000 was observed for this fraction. The sodium dodecyl sulfate-insoluble fraction could be dissolved upon reduction and alkylation and its composition was enriched in the amino acids characteristic of collagen. Polypeptides from this fraction were analyzed by electrophoresis in agarose and in agarose-acrylamide gels. The approximate molecular weight of the smallest component was 164,000. Additional polypeptides were observed whose molecular weights occurred in multimers of this component, up to 1.1 x 10(6), possibly indicating covalent cross-linked multimers of a basic collagen-like polypeptide(s).

Amino Acids↗
PubMed1979

Laser flow cytometric light scatter and fluorescence pulse width and pulse rise-time sizing of mammalian cells.

In laser flow cytometry, an increasingly popular technique of analytical cytology, quantitative measurements of interest include cell and nuclear diameters. Electronic circuitry for a new cell sizing technique has been developed which measured the time that signal pulses from either fluorescence or light scatter sensors exceed a preset constant fraction of the peak signal amplitude (pulse width) or the time that it takes a signal to rise between constant fractions of the peak signal amplitude on the rising side of the pulse (pulse rise-time). These pulse width or pulse rise-time measurements were related to cell or nuclear diameters and were used in combination to determine nuclear size to cell size ratios. This method of sizing was found to be independent of fluorescent or light-absorbing stain intensity, linearly related to cell or nuclear diameter, and capable of resolving small diameter differences.

Animals↗
PubMed1978

Electrokinetic alteration of the surface of herpes simplex virus infected cells.

Changes induced in the ionic composition of the surface of herpes simplex virus (HSV) infected cells were evaluated by whole cell electrophoresis and isoelectric focusing. The kinetics of the progressive decline in cell electrophoretic mobility (a measurement of negative surface charge density) after infection corresponded to the production of infection virus. The observed HSV-induced increase in cell isoelectric focusing pH demonstrated that the electrokinetic cell surface became ionically more positive. The nature of the ionic changes were investigated using a series of specific reagents in conjunction with cell electrophoresis. Infected cells had 2.4 times surface binding sites for the amino-specific reagent 4-acetamide-4'-isothiocyanostilbene-2,2'-disulphonic acid. A relationship between these virus-induced amino groups and HSV surface antigens was indicated by the similar kinetics of their appearance and electrophoretic neutralization by HSV antibodies.

Antigens, Viral↗