State Of Intracellular Water Research Articles

On the bioprotective effects of 3-hydroxybutyrate: Thermodynamic study of binary 3HB-water systems

Microorganisms must face various inconvenient conditions; therefore, they developed several approaches for protection. Such a strategy also involves the accumulation of compatible solutes, also called osmolytes. It has been proved that the monomer unit 3-hydroxybutyrate (3HB), which is present in sufficient concentration in poly(3-hydroxybutyrate) (PHB)-accumulating cells, serves as a chemical chaperone protecting enzymes against heat and oxidative stress and as a cryoprotectant for enzymes, bacterial cells, and yeast. The stress robustness of the cells is also strongly dependent on the behavior and state of intracellular water, especially during stress exposure. For a better understanding of the protective mechanism and effect of strongly hydrophilic 3HB in solutions at a wide range of temperatures, a binary phase diagram of system sodium 3HB (Na3HB)-water in equilibrium and the state diagrams showing the glass transitions in the system were constructed. To investigate the activity of water in various compositions of the Na3HB/water system, three experimental techniques have been used (dynamic water sorption analysis, water activity measurements, and sorption calorimetry). First, Na3HB proved its hydrophilic nature, which is very comparable with known compatible solutes (trehalose). Results of differential scanning calorimetry demonstrated that Na3HB is also highly effective in depressing the freezing point and generating a large amount of nonfrozen water (1.35 g of water per gram of Na3HB). Therefore, Na3HB represents a very effective cryoprotectant that can be widely used for numerous applications.

Molecular Dynamics Study of Effects of Temperature and Concentration on Hydrogen-Bond Abilities of Ethylene Glycol and Glycerol: Implications for Cryopreservation

The state of intracellular water is important in all phases of cryopreservation. Intracellular water can be transported out of the cell, transferred into its solid phase, or blocked by cryoprotectants and proteins in the cytoplasm. The purpose of the present study is to determine the amount of hydrogen-bonded water in aqueous ethylene glycol and glycerol solutions. The effects of temperature and concentration on the density and the hydrogen bonding characteristics of the solution are evaluated quantitatively in this study. To achieve these aims, a series of molecular dynamics simulations of ethylene glycol/water and glycerol/water mixtures of molalities ranging from 1 to 5 m are conducted at 1 atm and at 273, 285, and 298 K, respectively. The simulation results show that temperature and concentration have variable effects on solution density. The proportion of the hydrogen-bonded water by solute molecules increases with rising molality. The ability of the solute molecules to hydrogen bond with water molecules weakens as the solution becomes more concentrated. Moreover, it turns out that the solution concentration can influence the hydrogen bonding characteristics more greatly than the temperature. The glycerol molecule should be a stronger "water blocker" than the ethylene glycol molecule corresponding to the same conditions. These findings provide insight into the cryoprotective mechanisms of ethylene glycol and glycerol in aqueous solutions, which will confer benefits on the cryopreservation.

Characterization of High-pressure-induced Murine Erythroleukemia Cell Apoptosis by Proton Spin–Lattice Relaxation Times of Intracellular Water

Abstract In high-pressure-induced apoptosis, the state of intracellular water was examined by measuring proton spin–lattice relaxation time of water. At the early stage of apoptosis, bound water increased owing to water efflux, whereas free water increased due to its influx at the late stage.

Role of water in some biological processes

The state of intracellular water has been a matter of controversy for a long time for two reasons. First, experiments have often given conflicting results. Second, hitherto, there have been no plausible grounds for assuming that intracellular water should be significantly different from bulk water. A collective behavior of water molecules is suggested here as a thermodynamically inevitable mechanism for generation of appreciable zones of abnormal water. At a highly charged surface, water molecules move together, generating a zone of water perhaps 6 nm thick, which is weakly hydrogen bonded, fluid, and reactive and selectively accumulates small cations, multivalent anions, and hydrophobic solutes. At a hydrophobic surface, molecules move apart and local water becomes strongly bonded, inert, and viscous and accumulates large cations, univalent anions, and compatible solutes. Proteins and many other biopolymers have patchy surfaces which therefore induce, by the two mechanisms described, patchy interfacial water structures, which extended appreciable distances from the surface. The reason for many conflicting experimental results now becomes apparent. Average values of properties of water measured in gels, cells, or solutions of proteins are often not very different from the same properties of normal water, giving no indication that they are averages of extreme values. To detect the operation of this phenomenon, it is necessary to probe selectively a single abnormal population. Examples of such experiments are given. It is shown that this collective behavior of water molecules amounts to a considerable biological force, which can be equivalent to a pressure of 1,000 atm (1.013 x 10(5) kPa). It is suggested that cells selectively accumulate K+ ions and compatible solutes to avoid extremes of water structure in their aqueous compartments, but that cation pumps and other enzymes exploit the different solvent properties and reactivities of water to perform work of transport or synthesis.

Role of water in some biological processes.

The state of intracellular water has been a matter of controversy for a long time for two reasons. First, experiments have often given conflicting results. Second, hitherto, there have been no plausible grounds for assuming that intracellular water should be significantly different from bulk water. A collective behavior of water molecules is suggested here as a thermodynamically inevitable mechanism for generation of appreciable zones of abnormal water. At a highly charged surface, water molecules move together, generating a zone of water perhaps 6 nm thick, which is weakly hydrogen bonded, fluid, and reactive and selectively accumulates small cations, multivalent anions, and hydrophobic solutes. At a hydrophobic surface, molecules move apart and local water becomes strongly bonded, inert, and viscous and accumulates large cations, univalent anions, and compatible solutes. Proteins and many other biopolymers have patchy surfaces which therefore induce, by the two mechanisms described, patchy interfacial water structures, which extended appreciable distances from the surface. The reason for many conflicting experimental results now becomes apparent. Average values of properties of water measured in gels, cells, or solutions of proteins are often not very different from the same properties of normal water, giving no indication that they are averages of extreme values. To detect the operation of this phenomenon, it is necessary to probe selectively a single abnormal population. Examples of such experiments are given. It is shown that this collective behavior of water molecules amounts to a considerable biological force, which can be equivalent to a pressure of 1,000 atm (1.013 x 10(5) kPa). It is suggested that cells selectively accumulate K+ ions and compatible solutes to avoid extremes of water structure in their aqueous compartments, but that cation pumps and other enzymes exploit the different solvent properties and reactivities of water to perform work of transport or synthesis.

The information content of intracellular water and its accumulation in embryo- and phylogenesis (author's transl)

The nucleo-free erythrocytes is presented as the simplest differentiated cell whose energy-exchange has the single purpose of preserving the cell structure; this structure is based on the regular quasi-crystalline state of intracellular water, which is expressed by the negative entropy (-SW) and the temperature TW which is higher than the temperature of the extracellular water T. The information content of intracellular water, JW is proportional to the temperature difference: JW approximately delta T = TW - T. The regular state of intracellular water is maintained by the basal metabolism that takes place in the membrane of the differentiated cell. The energy exchange, i.e., the absorption of free energy and the liberation of an equivalent amount of heat, occurs in the form of work cycles of the enzyme-water-complexes in the cell membrane. The differentiated cell of the multicellular animal organism is the result of embryogenetic processes accompanied by heat-liberation. The specific heat-liberation, i.e., the heat produced by a single cell, begins with a quasi-zero value of the fertilized egg cell and grows with acceleration to a maximum value at the end of embryogenesis. This process of acceleration of heat-liberation is caused by the entrance of the water from the outer medium into the embryonic cell; the water undergoes the phase-change "fluid yields crystal" with heat-liberation. The intracellular water within the embryonic cell becomes structurated; this is also accompanied by growing heat-liberation. The thermodynamic characteristic of embryo genetic development is expressed by the principle of the maximum of velocity of entropy production of the cell at the end state of the process of differentiation. This principle applied to phylogenetics, leads to the formulation of the principle of accumulation of biological information: J(t). In the course of evolution living systems are able not only to store information of past generations, but also to create information: J(t) approximately e-1nt1!, where t is the time of phylogenesis.

State of water in extremely halophilic bacteria: Freezing transitions ofHalobacterium halobium observed by differential scanning calorimetry

The apparent latent heat of fusion (ΔH) of thick pastes ofHalobacterium halobium was determined by differential scanning calorimetry and compared with values obtained for a control paste of identical composition but containing bacteria already lysed by freezing and thawing. The rationale of the experiment was that, if there were any physico-chemical “abnormality” in the state of intracellular water or K+, it might be reflected in the freezing behavior of the two types of preparation. Differences were observed, the paste of whole cells consistently giving higher values of ΔH than the lysed bacteria. Furthermore, the differences between the two types of preparation increased with the age of the culture. Scans of whole-cell pastes apparently distinguished between intra- and extracellular water and enabled ΔH to be calculated for those two aqueous compartments. Supplementary measurements were also made on solutions of NaCl, KCl, and mixtures of the two salts. Sodium chloride lowered ΔH whereas KCl raised it. Differences in ΔH between internal and external solutions and between whole and lysed bacteria could be attributed to the different effects of the two salts on freezing behavior. Changing values of ΔH with the age of a culture correlated with changing ratios of K+/Na+ in the bacteria. It is concluded that there is no evidence at this level of any physicochemical abnormality in the halobacterial cytoplasm and no reason for supposing that retention of KCl by these bacteria is achieved by any agency other than the plasma membrane.

Osmotic properties of human lymphocyte.

The osmotic properties of human lymphocytes isolated from 15 ml of venous blood were examined. Measurements of the permeability of the membrane to water under an osmotic gradient were also made. The Boyle-Van't Hoff relation held very well for the human lymphocyte when the cells were shrunken in hyperosmotic media to concentrations twice isosmotic. The volume of osmotically inactive material or "b" value averaged 32% of the mean corpuscular volume. These values were independent of temperature. Ponder's R ranged between 0.8 and 0.9. The average value for Lp, the hydralic coefficient was 0.46 mu/min atm +/- 0.02 (S.E.M.) at 25 degrees C. No significant effect of age, sex, or race was noted. The effect of temperature between 10 degrees C and 37 degrees C was measured and heats of activation between 11.1 and 17.4 kcal/mole were calculated with a mean of 14.1 kcal/mole +/- 1.6 (S.E.M.). Concanavalin A at 10 microgram/1.5 X 10(6) lymphocytes produced blastogenesis of 25% or more of the lymphocytes without clumping, agglutination, or toxicity. The mean corpuscular volume increased by 21% after 72 hours due to an increase in the "b" value which increased by 80%. The volume of free water remained constant. Histograms of the distribution of cell volumes showed that volume changes were uniform throughout the population with no evidence of agglutination of clumping. The significance of these results is discussed in the context of membrane fluidity and the state of intracellular water.