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Biomedical subjects

F Franks

Publications and source records attributed to F Franks.

At least 37 records · Page 2Linked to original sources
PubMed1989

Variation in apparent enzyme activity in two-enzyme assay systems: phosphoenolpyruvate carboxylase and malate dehydrogenase.

We have employed the two-enzyme assay system for phosphoenolpyruvate to investigate the effect on the apparent phosphoenolpyruvate carboxylase (PEP-C) activity of the use of malate dehyrogenase (MDH) that has been stabilized in either glycerol or (NH4)2SO4. The type of MDH stabilizer has a marked effect on the apparent activity of the PEP-C. The apparent activities of the PEP-C are 1.34 and 0.43 U/mg in the presence of glycerol and salt-stabilized MDH, respectively. The implications of the observations for diagnostic assays are discussed.

Carboxy-Lyases↗
PubMed1988

The thermodynamics of protein stability. Cold destabilization as a general phenomenon.

A theoretical analysis of the temperature/stability profiles of proteins shows that, where a two-state model represents the denaturation, and where the free energy of denaturation delta G(T) shows a strong temperature dependence, then the protein becomes subject to both high- and low-temperature destabilization. In the simplest case delta G(T) is parabolic, therefore the high temperature TH, where delta (G(TH) = 0, is complemented by a low temperature TL, where delta G(TL) = 0. It is generally stated that the partial molal heat capacity change delta C accompanying the heat denaturation is positive and independent of the temperature. This implies that heating the protein through TL results in a negative delta C which seems physically unsatisfactory. The constant delta C model is explored and a physically more realistic model is advanced which allows for a temperature-dependent delta C which changes sign at some temperature within the range of stability of the native protein; delta G(T) then has the form of a skewed parabola. Experimental heat capacity data for native lysozyme and for a flexible polymer lend support to this model. The molecular basis of cold inactivation of proteins is discussed in the light of the thermodynamic analysis.

Cold Temperature↗
PubMed1986

Subzero-temperature preservation of reactive fluids in the undercooled state. II. The effect on the oxidation of ascorbic acid of freeze concentration and undercooling.

The rate of oxidation of ascorbic acid has been measured in both frozen and undercooled solutions. A new interpretation is advanced for changes in the rate of ascorbic acid oxidation in freeze-concentrated solutions. The results obtained with undercooled solutions indicate a rate reduction in line with that predicted by the Arrhenius equation. It is also demonstrated that undercoohng provides a method for greatly extending the shelf life of reactive fluids.

Journal Article↗
PubMed1986

Subzero-temperature preservation of reactive fluids in the undercooled state. I. The reduction of potassium ferricyanide by potassium cyanide.

Many reactions show enhanced rates at subzero temperatures due to freeze concentration. The reduction of potassium ferricyanide by potassium cyanide has been studied at subzero temperatures in both the undercooled and the frozen state. The pseudo-first-order rate constants calculated differ greatly from those in previous reports. A high degree of freeze concentration and supersaturation in frozen bulk solutions occurs. It has been clearly demonstrated that undercooled preservation provides a useful method for the long-term storage of reactive mixtures.

Journal Article↗
PubMed1985

Preservation of viable cells in the undercooled state.

Previous studies into the mechanisms governing the freezing of cells in the absence of extracellular ice have been extended to develop a method for the preservation of viable cells in the undercooled state. Deep undercooling of cells is achieved by suspending fine droplets of the cells in oil to make an emulsion, thus minimizing initiation of extracellular ice nucleation. Attempts to preserve yeast cells, cultured sainfoin cells, and dissected shoot-tips (pea and potato) in this way are described. The main findings are that yeast cells can be preserved undercooled at -20 degrees C for at least 16 weeks with no detectable loss of viability, showing that -20 degrees C is a low enough temperature for inhibition of significant biochemical deterioration and that the emulsions are stable over long periods. In preliminary experiments, sainfoin cells survived 24 hr at -10 degrees C, and shoot-tips survived 48 hr at -10 degrees C. Sainfoin cells, conditioned by growth in medium supplemented with sorbitol, showed enhanced survival after exposure to low temperatures and a lower intracellular freezing point than control cells. Possible reasons for this are discussed.

Animals↗
PubMed1984

Nucleation and growth of ice in deeply undercooled erythrocytes.

Previous studies of the mechanism of freezing of erythrocytes in the absence of intracellular ice have been extended to define the catalytic sites responsible for promoting nucleation. The following aspects have been investigated: (1) the freeze propagation between undercooled erythrocytes, (2) the nucleation of ice in undercooled erythrocyte ghosts, and (3) the freezing behavior of undercooled hemoglobin solutions. The main findings are: (1) no cross-nucleation occurs between individual cells packed within the same emulsified water droplet; (2) the differential scanning calorimetric power-time curves of intact cells and ghosts are identical, indicating that hemoglobin does not affect ice nucleation; (3) the nucleation temperature of ice in an aqueous solution of hemoglobin (isolated from the cells) is substantially lower than that for the same solution when contained in the intact cell; (4) the threefold freeze concentration which accompanies the freezing of a 25% hemoglobin solution does not cause denaturation of the protein.

Blood Preservation↗
PubMed1983

Ice nucleation and freezing in undercooled cells.

DSC has been employed to study the effect of cooling on a range of cells under exclusion of extracellular ice and in the absence of chemical cryoprotectants. In contrast to earlier reports, all the cells studied were found to freeze at temperatures above that indicated for homogeneous nucleation of ice in undercooled liquid water. In the case of human erythrocytes this temperature difference was only 0.5 degrees, but for yeast cells and cells of plant origin the difference amounted to congruent to 9 degrees. Nucleation of ice within the cell (or at the cell wall/membrane) must therefore be initiated by a heterogeneous mechanism. A kinetic analysis of the temperature dependence of nucleation shows the rates to be consistent with the dimensions of the plant cells (or organelles), if these were to be the active nucleators. However, the nucleation kinetics of human erythrocytes are extremely temperature sensitive, and the kinetic parameters only differ by small, though significant, extents from those of the suspension medium. Possible nucleation mechanisms are discussed in terms of the experimental data and the cell dimensions. Finally, one of the underlying assumptions of the kinetic analysis, i.e., that ice growth must be rapid compared to nucleation, has been tested and validated by freeze-fracture electron microscopy.

Calorimetry, Differential Scanning↗
PubMed1983

Solute--water interactions: do polyhydroxy compounds after the properties of water?

The interactions between PHCs and water, like those between water molecules, are governed by hydrogen bonding. The details of these interactions are very sensitive to spacings and orientations of the -OH groups on the solute molecules. Where different conformers can coexist in solution, the aqueous solvent acts so as to favor the conformer with the largest number of equatorial -OH groups, because of their spatial compatibility with water. Because of this compatibility, aqueous solutions of PHCs have the tendency to supersaturation and incomplete freezing, manifestations of the phenomenon known as bound water which is, however, a misnomer. The range of water structure perturbation is probably governed by hydration forces which appear to dominate at solute-solute distances of less than 3 nm and which decay exponentially. Although on a single hydrogen bond basis the hydration effects are marginal, they nevertheless are responsible for many macroscopic phenomena, e.g., gel formation, liquid crystals, and protection against dehydration.

Carbohydrates↗
PubMed1978

Blood glycoprotein from antarctic fish. Possible conformational origin of antifreeze activity.

High resolution 1H NMR and circular dichroism (CD) measurements have been performed on aqueous solutions of antarctic fish antifreeze glycoprotein. The carbohydrate contribution ot the observed CD spectrum has been estimated from closely analogous model compounds. The residual peptide contribution cannot be interpreted of the known spectral behaviour of alpha-helix, beta-sheet and random coil. Instead it resembles the CD spectrum of beta-structure in position, magnitude and spectral form, but is of opposite sign, indicating a specific but unusual peptide conformation, which we suggest may be stabilised by non-bonded interactions between the peptide backbone and the carbohydrate sidechains. Previous evidence which supports this interpretation is reviewed. NMR and CD measurements between -2 and +30 degrees C are consistent with conformational stability throughout the biologically relevant temperature range. The mechanism of the antifreeze activity is discussed in terms of the spatial and orientational correlations of sugar hydroxy groups and water in the liquid and solid states. The implication of an ordered peptide structure is explained by the comparison of the antifreeze glycoprotein with synthetic water-soluble polymers which also exhibit limited antifreeze properties.

Animals↗
PubMed1978

Supercooled water as medium for enzyme reactions at subzero temperatures.

A water-in-oil emulsion technique was employed to investigate enzyme-catalyzed reactions at sub-zero temperatures in the supercooled liquid state. The results obtained with a monooxygenase (bacterial cytochrome P-450) clearly indicate the potentialities as well as the technical problems of the procedure which might be successfully used to investigate enzyme systems sensitive to cosolvents.

Cold Temperature↗
PubMed1978

New trends in cryoenzymology: I.-Supercooled aqueous solutions.

A water in oil emulsion technique is proposed to investigate enzyme catalyzed reactions at sub-zero temperatures in the supercooled liquid state to avoid some reversible effects of the usual cosolvents on kinetics. Some results are listed: potentialities and technical problems of the procedure are discussed.

Alcaligenes↗
PubMed1977

Polymer cryoprotectants in the preservation of biological ultrastructure. I. Low temperature states of aqueous solutions of hydrophilic polymers.

The solid states formed by vitrified and frozen aqueous solutions of some hydrophilic polymers, able to act as biological cryoprotectants, have been studied by differential scanning calorimetry and freeze fracture electron microscopy. Glass transitions, devitrification, recrystallization and melting behaviour of aqueous solutions of polyvinylpyrrolidone, hydroxyethyl starch and dextran have been established. The vitrified polymer solutions exhibit a characteristic microspheral morphology which is not induced by the quench cooling process but is an inherent feature of the solutions themselves.

Cryoprotective Agents↗