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Biomedical subjects

K E Hunter

Publications and source records attributed to K E Hunter.

26 records · Page 2Linked to original sources

Retardation of embryogenesis by extremely low frequency 60 Hz electromagnetic fields.

Fertilized Medaka fish eggs were used to determine if electromagnetic fields, designed to simulate those beneath a high voltage power line, have biological effects on vertebrate embryo development. The newly fertilized eggs were exposed to a 60 Hz electrical field of 300 mA/m2 current density, a 60 Hz magnetic field of 1.0 gauss RMS, or to the combined electric plus magnetic fields for 48 hours. No gross abnormalities were observed in any of the embryos as they developed, but significant development delays were seen in those embryos exposed to either the magnetic or to the combined electromagnetic fields; delays were not seen in the embryos exposed to the electrical field. Thus, a 60 Hz magnetic field like that encountered in a man made powerline environment was shown to retard development of fish embryos.

Animals↗

Regulation of the permeability of the medaka fish embryo chorion by exogeneous sodium and calcium ions.

We questioned if the optically transparent noncellular chorion, or egg envelope, which encapsulates the entire medaka fish (Oryzias latipes) embryo might in some way constitute a permeability barrier to high concentrations of the diuretic called amiloride. More specifically, we questioned if removal of cations from the exogenous environment of the medaka embryo might make the chorion more permeable to amiloride and thereby make the fish embryos more sensitive to the inhibitory and lethal effects of this drug. To test this question, chorion-encapsulated medaka embryos were exposed to: deionized-distilled water, to Yamamoto-Ringer's (Y-R) solution, to Yamamoto-Ringer's containing choline chloride as a substitute for NaCl, and to isotonic NaCl solution in the presence of and in the absence of amiloride. Briefly, the prediction that the medaka embryos would be most sensitive to amiloride's inhibitory effects in distilled water was confirmed. Further studies showed that the presence of Na+ or of Ca2+ alone in the culture solution gave partial protection against the lethal effects of the amiloride. Electron probe X-ray microanalysis studies indicated that addition of Ca2+ and other cations to the culture solution caused the concentrations of cations to increase in the chorion, and that increase was correlated to a visible decrease in the permeability of the chorion to the amiloride. This decreased permeability of the chorion apparently protected the embryo from the amiloride. The decreased permeability of the chorion to amiloride, which occurred in the presence of the cations present in Y-R solution, was found to be reversible once the cations were washed from the chorion. Key words medaka, chorion, Na+, Ca2+ permeability, x-ray microanalysis, Oryzias latipes, egg envelope.

Amiloride↗

The subcellular concentration of ions and elements in thin cryosections of onion root meristem cells. An electron-probe EDS study.

Quantitative electron-probe energy dispersive X-ray microanalysis has, for the first time, been accomplished at a subcellular level in plant tissue using cryofixed and thin freeze-dried cryosections. The subcellular concentrations of Na+, Cl-, K+, P, S, Mg2+ and Ca2+ were measured in mol/kg dry weight in two types of root meristematic cells of the onion, Allium cepa. The cell wall of the meristematic cells had much higher concentrations of K+ and Ca2+ than was found in the intracellular compartments. Storage granules in the protoderm cells were about 6-12 times lower in P and were about four times higher in S as compared to other intracellular compartments. Comparison between the concentrations of ions and other elements in meristematic plant cells and in mouse cardiac myocytes confirms that major differences in cytoplasmic Na+ and Cl- concentrations do indeed exist between these cell types.

Animals↗

Nuclear-cytoplasmic distribution of ions in the hepatocyte.

To determine if nuclear/cytoplasmic (N/C) gradients of ions exist in mammalian hepatocytes, as has been shown to exist in the amphibian oocyte, small pieces of mouse liver were cryofixed for energy dispersive X-ray microanalysis of Na, K and Cl in thin freeze-dried cryosections. Expressing the data on a dry weight basis showed small but significant N/C gradients for Na and K but not for Cl. When the data were expressed on a wet weight basis no significant N/C gradients of Na, K or Cl were revealed. Questions of data interpretation are discussed.

Animals↗

Changes in water proton relaxation times and in nuclear to cytoplasmic element gradients during meiotic maturation of Xenopus oocytes.

Fully grown oocytes 1.2 mm in diameter were removed from Xenopus laevis ovaries and were exposed to progesterone (2.5 micrograms/ml in Ringer's solution) to induce completion of the first maturation division or germinal vesicle breakdown (GVBD). This process required 5.5 +/- 0.5 hr. Neither oocyte volume nor water content was observed to change throughout maturation. At selected times, the oocytes were quick frozen in liquid propane and cryosectioned. The sections were freeze-dried, and analyzed for K, Na, Cl, P, S, and Mg in millimolar per kilogram dry weight content in the nucleus and the yolk-free cytoplasm using electron probe X-ray microanalysis. Unstimulated oocytes showed significant nuclear to yolk-free cytoplasmic content gradients (N/C ratio) for the following elements: K (1.84), P (0.65), and S (1.56), but significant N/C content gradients were not found for Na and Mg. By 10 min after progesterone stimulation, a significant change in the N/C ratio of the following elements had occurred due to a rapid increase in nuclear content: K (2.29), Cl (2.11). A significant N/C ratio for Mg (1.35) had developed by 10 min after progesterone stimulation and a significant N/C ratio for Na (2.07) had developed by 45 min. In addition the following elements showed significant content increases in both the nucleus and the yolk-free cytoplasm from the time prior to progesterone stimulation to the time just prior to GVBD at 240 min: K, Na, Cl, P, S, and Mg. Nuclear magnetic resonance measurements of the spin-lattice relaxation time (T1) of water proton in oocytes showed a significant increase in the T1 time after progesterone exposure. The changes in N/C ratios of specific elements and in the physical parameter of water proton relaxation time suggest that progesterone is responsible for inducing changes in the physicochemical interactions between various macromolecules, specific elements, and water.

Animals↗

Change in water proton relaxation time during erythrocyte maturation.

Erythrocyte populations from newborn and mature mice were characterized according to: size; ultrastructural features; water content; concentration of intraerythrocyte elements including Na, Cl, K, P, S, Mg, and Fe; and the spinlattice (T1) and spin-spin relaxation times of water protons as measured by nuclear magnetic resonance (NMR) spectroscopy. A significant increase in the T2 time from 142 +/- 3 msec to 184 +/- 3 msec occurred during erythrocyte maturation. This change in T2 time was correlated with a change from a polyribosome-rich hemoglobin-poor cell type to a polyribosome-absent hemoglobin-rich cell type. The change in T2 time could also be correlated to a significantly higher K and P concentration in the mature erythrocytes. The change in T2 time was not correlated to a change in cellular water content or to the concentration of any of the other elements measured by electron probe X-ray microanalysis. If the NMR relaxation times of water molecules truly reflect their average motional freedom, then the findings suggest that greater water ordering interaction occurs in the ribosome containing immature erythrocyte.

Animals↗

Effect of cancer cachexia and amiloride treatment on the intracellular sodium content in tissue cells.

This study was designed to investigate the effects of a growing H6 hepatoma on the intracellular element content in three distinctly different tissue cell populations of the mouse host (hepatocytes, fibroblasts, and crystal enterocytes). X-ray microanalysis measurements of the intranuclear concentrations of several elements (sodium, magnesium, phosphorus, sulfur, chlorine, and potassium) were made. Briefly, the tumor presence significantly increased intranuclear sodium concentration but not the concentration of magnesium, phosphorus, sulfur, chlorine, or potassium in three tissue cell types of mice that were anorectic and cachectic. A second aim of the study was to see if injections of the diuretic amiloride, a drug reported to block passive influx of sodium into mammalian cells, would counteract the effect of the tumor presence and lower the intranuclear concentration of sodium towards that of a non-tumor-bearing host. Amiloride did significantly lower the intranuclear level of sodium in the host tissues to that of non-tumor-bearing mice. The amiloride-caused decrease on intracellular sodium was correlated to a decreased cell proliferation activity in the tumor cells and duodenal enterocytes. A possible relationship between the intracellular concentration of sodium in tissue cells and cancer cachexia is discussed.

Amiloride↗