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Biomedical subjects

I L Cameron

Publications and source records attributed to I L Cameron.

At least 91 records · Page 5Linked to original sources
PubMed1986

Elemental concentration gradients between subcellular compartments.

To gain information on the mechanisms involved in the establishment and maintenance of subcellular gradients of Na, K, Cl and other elements in the flagellate, Euglena gracilis, we turned to the technique of ultracentrifugal stratification of its intracellular contents, which is achieved without loss of viability or cell rupture. Stratified and non-stratified Euglena were cryofixed for energy-dispersive X-ray microanalysis of Na, K, Cl and other elements in thin freeze-dried cryosections. A number of significant elemental concentration differences (expressed as mmol kg-1 dry weight) were found between chloroplast, nucleus, paramylon granules and open cytoplasm (which contained ribosomes, membranes and macromolecules associated with the cytomatrix) in the non-stratified cells. Stratification caused several ions to be redistributed. For example, we observed a significant increase in K and Cl in the nucleus, which was correlated with the condensation of chromatin. Also Cl, but not Na, decreased significantly in the region of cytoplasm that was cleared of observable ribosomes, membranes and macromolecules associated with the cytomatrix, as well as of observable cytochemical enzyme activity. We conclude from the data that more than half of the Cl in open cytoplasm was adsorbed to or entrapped in material that was removed by ultracentrifugation. Thus, it appears that a close association of at least one ion, Cl, with ultracentrifugable material is involved in maintenance of the measured Cl concentration in the open cytoplasm of the non-stratified cell.

Animals↗
PubMed1986

Disruption of the plasma membrane leads to an influx of Na+ and Cl- but not an efflux of K+ in mouse hepatocytes and erythrocytes.

To determine what mechanisms might be involved in the maintenance of the known extra-/intracellular concentration gradients of Na+, Cl- and K+, small pieces of mouse liver and heparinized blood were appropriately cryofixed. The tissues were cryosectioned and cryosorbed at -100 degrees C or at -40 degrees C. The former temperature prevented diffusion of all ions as measured by electron probe x-ray microanalysis of the thin (0.1 micron) cryosorbed sections of the cells while the latter temperature allowed significant diffusion of Na+ and Cl- into the hepatocytes and erythrocytes but did not allow diffusion of K+ from the hepatocytes or the erythrocytes. These results indicate that the plasma membrane is involved in maintenance of the extra-/intracellular gradients of Na+ and Cl- but that intracellular association of K+ with macromolecules is the main mechanism responsible for maintenance of the extra-/intracellular K+ concentration gradient.

Animals↗
PubMed1985

Orientation of tendons in the magnetic field and its effect on T2 relaxation times.

In vitro studies of spin-spin relaxation times of the protons or hydrogen nuclei which make up the tendon demonstrated significant dependence of T2 on orientation at physiological levels of hydration. T2 varied from approximately 250 mu sec. with the tendon aligned with the magnetic field to 22 msec. at an angle of 55 degrees or 125 degrees and 4 msec. at deviations of +/- 15 degrees from 55 degrees and 125 degrees. 55 degrees and 125 degrees are the so-called "magic angles" at which 3 cos2 theta - 1 = 0. As a result, the signal from human tendons in vivo will depend on their orientation; however, T2 is generally short enough that the signal intensity will approach 0 in MR images regardless of the orientation of the tendons, thereby preventing this phenomenon from being observed in most studies.

Animals↗
PubMed1985

Proton magnetic resonance relaxation behavior of whole muscle with fatty inclusions.

This in vitro proton study of spin lattice (T1) and spin spin (T2) relaxation of muscle with storage-fat inclusions demonstrates slow exchange and lack of cross-relaxation between fat and water. Slow exchange causes biphasic T1 relaxation, but T2 relaxation is paradoxically uniphasic due to the nearly equal T2 values for both fractions. By careful dehydration and fat extraction, the relaxation information was deconvolved into water, fat, and protein contributions. The biphasic T1 decay has a short component due to lipid and a long component due to the water-protein combination. The fat content of muscle can be measured from the relative amplitude of the two T1 components or directly from the T2 relaxation time.

Adipose Tissue↗
PubMed1985

Water ordering during the cell cycle: nuclear magnetic resonance studies of the sea-urchin egg.

Nuclear magnetic resonance was used to measure spin-lattice water proton relaxation times (T1) during the first cell cycle in sea-urchin zygotes of packed Strongylocentrotus purpuratus. Following insemination there was a 90% increase in the T1 value. The increase in T1 at fertilization could be accounted for by the accumulation of extracellular fluid between the egg surface and the fertilization envelope. The T1 value then remained without change during the first cell cycle, except at metaphase when there was a significant 13% decrease. The lowered T1 values measured at metaphase were not related to a change in the water content of the packed cells, which remained fairly constant throughout the cell cycle. High hydrostatic pressure, low temperature and colchicine (agents that depolymerize mitotic apparatus microtubules) did not affect the T1 values in fertilized eggs. Treatment in vitro of a microtubule protein preparation with low temperature and colchicine resulted in an increased T1, which accompanied the depolymerization of microtubule protein. Since depolymerization of the microtubules associated with the mitotic apparatus by high pressure, colchicine or low temperature does not alter the T1 of water protons in the cell, it is proposed that the increased state of ordered water molecules at metaphase is maintained by nonmicrotubular factor(s) of the metaphase egg.

Animals↗
PubMed1985

Subcellular flux of potassium and rubidium in amphibian oocytes.

To determine the net Rb+ influx and K+ efflux at a subcellular level, fully grown and a Ringer's solution where K+ was substituted for by Rb+ on a molar bases. For 40 hrs serial samples of the oocytes were cryofixed and cryosectioned for elemental analysis in mM per kg dry weight using electron probe x-ray microanalysis. Oocyte volume remained constant. Net Rb+ influx showed a slow exponential increase into the nucleus, the yolk-free cytoplasm and the yolk platelets. There was significant K+ efflux from the nucleus but not from the yolk-free cytoplasm or the yolk. The Na+ concentration remained unchanged in all compartments during the course of the experiment. There was however a slow but significant increase in the concentration of Cl- in each of the subcellular compartments but this increase was not sufficient to balance the observed increase in the sum of K+ plus Rb+. Thus Rb+ accumulates selectively in all three subcellular compartments indicating that Rb+ is not a good K+ surrogate in the oocytes. That K+ demonstrates efflux from the nucleus but not from the cytoplasmic compartments is interpreted to suggest that some of the nuclear K+ is lost in exchange for Rb+ but that essentially none of the cytoplasmic K+ is-lost in exchange for Rb+. The findings provide strong evidence for adsorption of Rb+ in the cell.

Animals↗
PubMed1985

Relationships between ice crystal size, water content and proton NMR relaxation times in cells.

Biological specimens were frozen under controlled conditions. We questioned how the size of ice crystals, as measured in cryosectioned and cryoadsorbed sections of these biological specimens, relates to the water content and to the proton NMR relaxation times (T1 and T2) of the unfrozen specimens. The results permit the following conclusions: After rapid freezing in liquid propane cooled in a liquid nitrogen bath, the average size of ice crystals at distances of 150 microns or more from the surface of a particular tissue was always the same. Thus, the average size of the ice crystals was found to be characteristic of the type of biological tissue studied. Linear regression analysis showed average ice crystal size to have a significant correlation coefficient to T1 relaxation time and to water content. Specifically ice crystal size increased with T1 relaxation time and with water content. Multiple regression and path analysis demonstrated a positive correlation between the T1 relaxation time and the ice crystal size variation. Path analysis showed that both water content and T2 relaxation time were less directly correlated with ice crystal size. The findings from the path analysis and other observations show that the average size of ice crystals in subcellular compartments is best predicted by the proton T1 relaxation time. A working model is put forth to explain differences in ice crystal size observed between specimens enriched in globular or in parallel filamentous proteins.

Animals↗
PubMed1985

Electron probe x-ray microanalysis studies on the ionic environment of nuclei and the maintenance of chromatin structure.

We have demonstrated the application of electron probe X-ray microanalysis to studies aimed at determining the role of the ionic environment in the maintenance of a stable chromatin structure during interphase. Our results indicate that X-ray microanalysis can be used to accurately measure the concentration of ions, such as Na+ and K+ within subcellular compartments. We also present evidence from our X-ray microanalysis studies to show that only a portion of the K+ in the interphase nucleus is in a free ionic state and that the rest of the intranuclear K+ is adsorbed to intranuclear macromolecules. The stable structural state of interphase chromatin was correlated with the maintenance of a normal range in the chemical activities of monovalent ions in the oocyte nucleus. Finally, we demonstrated that the chemical activity of the monovalent ions probably increases beyond this normal interphase range as the chromatin in the nucleus condenses into discrete chromosomes during the first meiotic division in the amphibian oocyte. The above observations support the hypothesis of Kellermayer et al. (1984) that the maintenance of chromatin structure in the nucleus of living cells requires a balance between the presence or absence of mobile nuclear proteins and the chemical activity of monovalent cations. It also suggests that an elevation of the chemical activity of the monovalent cations above a normal range will result in the destabilization of interphase chromatin structure. It is hypothesized that an elevation of monovalent cations may be directly involved in the structural and functional changes in chromatin which are known to take place at the time of meiosis and mitosis.

Animals↗
PubMed1985

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↗
PubMed1984

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↗
PubMed1984

Characterization of proton NMR relaxation times in normal and pathological tissues by correlation with other tissue parameters.

To help understand which tissue parameters best account for the water proton NMR relaxation times, the longitudinal relaxation time (T1), the transverse relaxation time (T2), and the water content of 16 tissues from normal adult rats were measured at 10.7 MHz and 29 degrees C. Regression analyses between the above and other tissue parameters were performed. These other tissue parameters included: the amounts of various organic and inorganic components, protein synthetic rate, oxygen consumption rate, and morphological composition. In addition, the differences in T1, T2, and water content values between normal liver and malignant tumor (Morris #7777 a transplantable hepatoma) were studied to help understand how a disease state can be detected and characterized by NMR spectroscopy. The results of this study and information from the literature allow the following generalizations to be made about tissue T1 and T2 values: (1) Each normal tissue has rather consistent and characteristic T1 and T2 relaxation times which are always shorter than the T1 and T2 of bulk water; (2) tissues with higher water content tend to have longer T1 relaxation times; (3) tissue T2 values are not, however, as well correlated with water content as T1 values; (4) tissues with shorter T1 values have higher calculated hydration fractions, greater amounts of rough endoplasmic reticulum, and a greater rate of protein synthetic activity; (5) tissues with higher lipid content, associated with intracellular non-membrane bounded lipid droplets, tend to have longer T2 values; (6) tissues with greater overall surface area, whether in the form of cellular membranes or intracellular or extracellular fibrillar macromolecules, tend to have shorter T2 values; (7) the differences between T1 and T2 values between tumor and normal tissues correlated with differences in the volume fraction (amounts) of extracellular fluid volumes and in the amounts of membrane and fibrillar surface area in the cells. The above generalizations should be useful in predicting T1 and T2 changes associated with specific tissue pathologies.

Amino Acids↗
PubMed1984

Amiloride and other blockers of electrolyte flux as inhibitors of progesterone-stimulated meiotic maturation in Xenopus oocytes.

Amiloride and seven of its analogues, as well as seven substances of other structural types, all of which control electrolyte transport, were tested for their effectiveness for inhibiting the progesterone-stimulated meiotic maturation cycle of oocytes from the amphibian Xenopus laevis. Data were also collected on the ability of the oocytes to recover from the drug's inhibitory effect and on the toxicity of each drug. The data revealed that up to a 14-fold difference exists in the inhibitory concentration 50% between substances, that recovery from the drug's inhibitory effect ranged from non-reversible to almost complete reversibility and that toxicity, as measured by failure to exclude trypan blue, ranged from 0 to 57%. In some, but not all cases, the failure to recover from the drug's inhibitory effects could be correlated to the drug's toxic effects. Amiloride which has been shown to be a reversible inhibitor of cell proliferation in rapidly dividing mammalian cell populations has similar properties on the oocyte maturation division cycle of Xenopus.

Amiloride↗
PubMed1984

Frequency dependence of magnetic resonance spin-lattice relaxation of protons in biological materials.

The frequency dependence of spin-lattice (T1) relaxation times of protons in tissues and other macromolecular solutions was investigated. This dependence can be principally assigned to reduced mobility of water molecules "hydrated" on the surface of macromolecular structures. The T1 relaxation time of "hydration water" was found to increase linearly with frequency in the range from 5 to 100 MHz (T1 = 1.83 f + 25.0). It is assumed that the remainder of water in the tissue has a relaxation rate that is independent of frequency, as is characteristic of bulk tap water. Variations occur in the fraction of water hydrated or bound from one organ to the next. As the observed relaxation rate is a weighted average of the two rates as described by the fast exchange model, the above empirical relationship can be used to take a tissue T1 relaxation time measured at one frequency and then calculate the tissue T1 relaxation time that would be expected at another frequency. Good agreements were obtained between such calculated tissue T1 values and the T1 values actually measured at the second frequency. Variations between the calculated and measured T1 values in some classes of tissues indicate that there are also less important secondary factors such as lipid content.

Achilles Tendon↗
PubMed1984

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↗
PubMed1984

Post mortem changes in skeletal muscle proton spin-lattice relaxation times.

A previously published report indicated that there are early post mortem changes in the pulsed NMR proton spin-lattice relaxation time (T1) of skeletal muscle which must be taken into account in in vitro tissue analysis. We re-examined this subject. When T1 measurements were done by allowing the signal intensity to decay over two orders of magnitude from the original intensity, the T1 decay curves showed more than a single exponential relaxation time component as reported earlier. However, when T1 measurements were done by allowing signal intensity to decay to about one order of magnitude from the original intensity only a single exponential relaxation time component was found. We postulate that the latter method gives the average T1 value of the various macroscopic tissue components present and that this method gives a representative T1 value for the entire tissue. Using data obtained in this manner we could not find significant post mortem changes in the muscle T1 relaxation times during the first four hours but found change at a later time.

Animals↗
PubMed1984

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↗
PubMed1983

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↗