THE SWELLING OF ISOELECTRIC GELATIN IN WATER : I. EQUILIBRIUM CONDITIONS.

The swelling of isoelectric gelatin in water has been found to be in agreement with the following assumptions. Gelatin consists of a network of insoluble material containing a solution of a more soluble substance. Water therefore enters owing to the osmotic pressure of the soluble material and thereby puts the network under elastic strain. The process continues until the elastic force is equal to the osmotic pressure. If the temperature is raised or the blocks of gelatin remain swollen over a period of time, the network loses its elasticity and more water enters. In large blocks this secondary swelling overlaps the initial process and so no maximum can be observed. The swelling of small blocks or films of isoelectric gelatin containing from .14 to .4 gm. of dry gelatin per gm. of water is defined by the equation See PDF for Equation in which K_e = the bulk modulus See PDF for Equation. V_e = gm. water per gm. gelatin at equilibrium; V_f = gm. water per gm. gelatin when the gelatin solidified.     

INCREASED PERMEABILITY TO WATER OF AGING UNFERTILIZED EGGS (ARBACIA PUNCTULATA)

1. Unfertilized Arbacia eggs, subjected to 50, 60, or 80 per cent sea water, at the same temperature, pH, and duration, swelled progressively faster with age. Swelling increased 12.2 to 39.6 per cent. 2. Permeability to water was calculated by the method of Lucké and McCutcheon. For freshly shed eggs from nine different females, permeability varied from 0.078 to 0.124, or 59 per cent. This variation was not due to differences in age nor to physiologic condition but was correlated with size of eggs. 3. About twenty eggs were tested each age to 60 hours after shedding. The permeability rate increased with age in all nine experiments. 4. The total increase ranged from 30 to 69 per cent. 5. The increase was slow but constant or almost constant during early and intermediate ages. 6. The increase began at earliest ages. 7. An increase may not be demonstrable for any short interval but is readily demonstrable for longer intervals (6 or more hours), or, from the entire curve. 8. Permeability increased faster at late ages. For special factors during late ages see text. 9. Injury by experimental procedure was not induced until ca. the 24th hour. Injury by aging was not detectable during the first 6 to 9 hours. It increased progressively between 10 and 24 hours, and very rapidly thereafter. Permeability increased at approximately the same rate during early and intermediate ages. Permeability therefore was not correlated with degree of injury. 10. The troublesome factor in the permeability equation was initial size, or V_o. Several methods were used to obtain this V_o, viz. (a) extrapolations approximating known cyclical change in size, (b) extrapolations approximating control sizes, (c) constant or average of all V_o''s, (d) maximum V_o approximating control V_o''s. Recalculation with these different V_o''s gave approximately the same permeabilities. 11. The increase in permeability with age was substantially the same whether "approved," or other possible rates or average rates or range of rates be used as criteria of change. 12. When the data were recalculated by integrating the equation of Northrop, the increases in permeability with age were essentially the same as the increases calculated by the graphic method of Lucké.     

SYNERESIS AND SWELLING OF GELATIN

1. When solid blocks of isoelectric gelatin are placed in cold distilled water or dilute buffer of pH 4.7, only those of a gelatin content of more than 10 per cent swell, while those of a lower gelatin content not only do not swell but actually lose water. 2. The final quantity of water lost by blocks of dilute gelatin is the same whether the block is immersed in a large volume of water or whether syneresis has been initiated in the gel through mechanical forces such as shaking, pressure, etc., even in the absence of any outside liquid, thus showing that syneresis is identical with the process of negative swelling of dilute gels when placed in cold water, and may be used as a convenient term for it. 3. Acid- or alkali-containing gels give rise to greater syneresis than isoelectric gels, after the acid or alkali has been removed by dialysis. 4. Salt-containing gels show greater syneresis than salt-free gels of the same pH, after the salt has been washed away. 5. The acid and alkali and also the salt effect on syneresis of gels disappears at a gelatin concentration above 8 per cent. 6. The striking similarity in the behavior of gels with respect to syneresis and of gelatin solutions with respect to viscosity suggests the probability that both are due to the same mechanism, namely the mechanism of hydration of the micellæ in gelatin by means of osmosis as brought about either by diffusible ions, as in the presence of acid or alkali, or by the soluble gelatin present in the micellæ. The greater the pressures that caused swelling of the micellæ while the gelatin was in the sol state, the greater is the loss of water from the gels when the pressures are removed. 7. A quantitative study of the loss of water by dilute gels of various gelatin content shows that the same laws which have been found by Northrop to hold for the swelling of gels of high concentrations apply also to the process of losing water by dilute gels, i.e. to the process of syneresis. The general behavior is well represented by the equations: See PDF for Equation and See PDF for Equation where P ₁ = osmotic pressure of the soluble gelatin in the gel, P ₂ = stress on the micellæ in the gelatin solution before setting, K_e = bulk modulus of elasticity, V_o = volume of water per gram of dry gelatin at setting and V_e = volume of water per gram of gelatin at equilibrium.     

KINETICS OF THE SWELLING OF CELLS AND TISSUES

The rate of swelling of Arbacia eggs in dilute sea water, studied by Lillie and by Lucke and McCutcheon, may be expressed by the formulæ derived for the rate of increase in volume of a solution enclosed in a collodion sac. The rate of swelling of slices of carrot in distilled water, measured by Stiles and Jørgensen, may be expressed by the equation derived previously for the swelling of similarly shaped blocks of gelatin.     

Water transport in invertebrate peripheral nerve fibers

Osmotic and diffusion permeabilities (P_f and P_d) of invertebrate nerve fibers to tritiated water were measured to determine what water flux studies could reveal about "the nerve membrane" and to directly test the possibility of active transport of water into or out of invertebrate nerve fibers. P_f/P_d ratios for lobster walking leg nerve fibers were found to be about 20 ± 7 at 14°C. P_d measurements were made for squid giant axons at 25°C. and found to yield a value of 4 x 10^–4 cm.^–1 sec.^–1. When combined with the data of D. K. Hill for P_f, a P_f/P_d ratio of 21 ± 5 is obtained. These P_f/P_d ratios correspond to "effective pore radii" of about 16 ± 4 angstrom units, according to theories developed by Koefoed-Johnsen and Ussing and independently by Pappenheimer and his colleagues. Variations of water flux ratios with temperatures were studied and apparent activation energies calculated for both diffusion experiments and osmotic filtration experiments using the Arrhenius equation, and found to be close to 3 to 5 cal. per mole of water transferred. Cyanide (5 x 10^–3 molar) and iodoacetate (1 x 10^–3 molar) poisoned lobster leg nerve fibers showed no appreciable change in diffusion or osmotic filtration water effluxes. Caution in interpreting these proposed channels as simple pores was emphasized, but the possibility that such channels exist and are related to ionic flow is not incompatible with electrophysiological data.     

Kinetics of erythrocyte swelling and membrane hole formation in hypotonic media

Red blood cell (RBC) swelling and membrane hole formation in hypotonic external media were studied by measuring the time-dependent capacitance, C, and the conductance, G, in the beginning of the β-dispersion range. At high and moderate osmolarities of the external solution the capacitance reaches a steady-state whereas at low osmolarities it reveals a biphasic kinetics. Examination of RBC suspensions exposed to different concentrations of HgCl₂ demonstrates that water transport through mercury-sensitive water channel controls RBC swelling. Unlike the capacitance, an increase in the conductance to a stationary level is observed after a certain delay. A comparison of G(t) curves recorded for the suspensions of the intact cells and those treated with cytochalasin B or glutaraldehyde demonstrates the significant effect of the membrane viscoelasticity on the pore formation. It is shown that the stretched membrane of completely swollen RBC retains its integrity for a certain time, termed as the membrane lifetime, t_memb. Therefore, the resistivity of RBCs to a certain osmotic shock may be quantified by the distribution function of RBC(t_memb).     

OSMOTIC PROPERTIES OF THE EGG CELLS OF THE OYSTER (OSTREA VIRGINICA)

Investigations of the osmotic properties of oyster eggs by a diffraction method for measuring volumes have led to the following conclusions: 1. The product of cell volume and osmotic pressure is approximately constant, if allowance is made for osmotically inactive cell contents (law of Boyle-van''t Hoff). The space occupied by osmotically inactive averages 44 per cent of cell volume. 2. Volume changes over a wide range of pressures are reversible, indicating that the semipermeability of the cell during such changes remains intact. 3. The kinetics of endosmosis and of exosmosis are described by the equation, See PDF for Equation, where dV is rate of volume change; S, surface area of cell, (P-P_e), the difference in osmotic pressure between cell interior and medium, and K, the permeability of the cell to water. 4. Permeability to water during endosmosis is 0.6µ³ of water per minute, per square micron of cell surface, per atmosphere of pressure. The value of permeability for exosmosis is closely the same; in this respect the egg cell of the oyster appears to be a more perfect osmometer than the other marine cells which have been studied. Permeability to water computed by the equation given above is in good agreement with computations by the entirely different method devised by Jacobs. 5. Permeability to diethylene glycol averages 27.2, and to glycerol 20.7. These values express the number of mols x 10^–15 which enter per minute through each square micron of cell surface at a concentration difference of 1 mol per liter and a temperature of 22.5°C. 6. Values for permeability to water and to the solutes tested are considerably higher for the oyster egg than for other forms of marine eggs previously examined. 7. The oyster egg because of its high degree of permeability is a natural osmometer particularly suitable for the study of the less readily penetrating solutes.     

Volume regulation in the coelomocytes of the blood wormGlycera dibranchiata

Summary Median volumes ofin vitro coelomocyte populations fromGlycera dibranchiata rapidly change in response to external differences of osmotic pressure (Fig. 3). Fifty per cent haemolysis occurred in just under 30% sea water, 285 mOSm·kg^–1. Hydraulic conductivities (L_p=0.92 to 2.78×10⁷ cm×s^–1·atm^–1) calculated from rates of osmotic swelling were similar to values for sea urchin eggs and squid axons. Coelomocytes show a slower partial return to their original volumes in hypotonic but not hypertonic media. This asymmetry is reflected in Ponder's R values of 0.787 and 0.987, respectively, determined after this regulatory phase is complete (Figs. 4 and 5). Evidence for an irreversible stress dependent leakage of osmotically active solutes when the coelomocytes are removed from the animal and diluted with sea water is presented.     

Photosynthetic limitations of a halophyte sea aster (<Emphasis Type="Italic">Aster tripolium</Emphasis> L) under water stress and NaCl stress

To understand the mechanisms of salt tolerance in a halophyte, sea aster (Aster tripolium L.), we studied the changes of water relation and the factors of photosynthetic limitation under water stress and 300 mM NaCl stress. The contents of Na⁺ and Cl^- were highest in NaCl-stressed leaves. Leaf osmotic potentials (Ψ _s) were decreased by both stress treatments, whereas leaf turgor pressure (Ψ _t) was maintained under NaCl stress. Decrease inΨ _s without any loss ofΨ _t accounted for osmotic adjustment using Na⁺ and Cl^- accumulated under NaCl stress. Stress treatments affected photosynthesis, and stomatal limitation was higher under water stress than under NaCl stress. Additionally, maximum CO₂ fixation rate and O₂ evolution rate decreased only under water stress, indicating irreversible damage to photosynthetic systems, mainly by dehydration. Water stress severely affected the water relation and photosynthetic capacity. On the other hand, turgid leaves under NaCl stress have dehydration tolerance due to maintenance of Ψ _t and photosynthetic activity. These results show that sea aster might not suffer from tissue dehydration in highly salinized environments. We conclude that the adaptation of sea aster to salinity may be accomplished by osmotic adjustment using accumulated Na⁺ and Cl^-, and that this plant has typical halophyte characteristics, but not drought tolerance. Electronic Publication     

Evaluation of the importance of ionic and osmotic components of salt stress on the photosynthetic efficiency of epiphytic lichens

Salt stress can significantly disrupt the functioning of lichens which are self-sufficient symbiotic organisms inhabiting various severe environments. The aim was to test the effect of salt and sucrose on the photosynthetic efficiency of two selected epiphytic lichens inhabiting the interior of the land. Firstly, we compared the effect of salt and sucrose solutions of different concentrations. Secondly, the effect of salt and sucrose solutions with identical osmotic pressures was compared. The results showed that short-term salt stress leads to a significant reduction of F_V/F_M, greater changes in chlorophyll fluorescence parameters and OJIP transients compared to the osmotic effects induced by sucrose. This proved that the negative impact of salt stress is associated primarily with ionic effects. The most symptomatic effect of the ionic stress was a significant reduction of the utilisation of trapped energy in electron transport and thereby down-regulation of electron transfer. Since lichens are resistant to a temporary lack of water, ionic stress could have more serious consequences than osmotic stress itself. Hypogymnia physodes was more sensitive to salt stress than Pseudevernia furfuracea, but the reduction of photosynthetic efficiency was not permanent since after 24 h F_V/F_M returned to the level characteristic for healthy lichens. Nevertheless, repeated exposure to salt may reduce the vitality of lichens growing along communication routes sprinkled with salt in the winter season. Finally, the changes in certain JIP-test parameters were stronger than F_V/F_M, thus they could be better indicators of salt stress in lichens.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-022-01134-2.     

Increased Hepato-Splanchnic Vasoconstriction in Diabetics during Regular Hemodialysis

Background and Objectives

Ultrafiltration (UF) of excess fluid activates numerous compensatory mechanisms during hemodialysis (HD). The increase of both total peripheral and splanchnic vascular resistance is considered essential in maintaining hemodynamic stability. The aim of this study was to evaluate the extent of UF-induced changes in hepato-splanchnic blood flow and resistance in a group of maintenance HD patients during regular dialysis.

Design, Setting, Participants, & Measurements

Hepato-splanchnic flow resistance index (RI) and hepato-splanchnic perfusion index (QI) were measured in 12 chronic HD patients using a modified, non-invasive Indocyaningreen (ICG) dilution method. During a midweek dialysis session we determined RI, QI, ICG disappearance rate (k _ICG), plasma volume (V _p), hematocrit (Hct), mean arterial blood pressure (MAP) and heart rate (HR) at four times in hourly intervals (t ₁ to t ₄). Dialysis settings were standardized and all patient studies were done in duplicate.

Results

In the whole study group mean UF volume was 1.86 ± 0.46 L, V _p dropped from 3.65 ± 0.77L at t₁ to 3.40 ± 0.78L at t₄, and all patients remained hemodynamically stable. In all patients RI significantly increased from 12.40 ± 4.21 mmHg∙s∙m²/mL at t₁ to 14.94 ± 6.36 mmHg∙s∙m²/mL at t₄ while QI significantly decreased from 0.61 ± 0.22 at t₁ to 0.52 ± 0.20 L/min/m² at t₄, indicating active vasoconstriction. In diabetic subjects, however, RI was significantly larger than in non-diabetics at all time points. QI was lower in diabetic subjects.

Conclusions

In chronic HD-patients hepato-splanchnic blood flow substantially decreases during moderate UF as a result of an active splanchnic vasoconstriction. Our data indicate that diabetic HD-patients are particularly prone to splanchnic ischemia and might therefore have an increased risk for bacterial translocation, endotoxemia and systemic inflammation.     

Kinetic models for the dynamical behavior of polyacrylamide (PAAm)–κ-carrageenan (κC) composite gels

A fluorescence method was employed for studying the drying and swelling of PAAm–κC composite gels, which were formed from acrylamide (AAm) and N, N’- methylenebisacrylamide (BIS) with various κ–carrageenan (κC) contents by free radical crosslinking copolymerization in water. Composite gels were prepared at 80 °C with pyranine (Py) as a fluorescence probe. Scattered light, I_sc, and fluorescence emission intensities, I_em, were monitored during drying and swelling of these gels. The fluorescence intensity of pyranine increased and decreased as drying and swelling time are increased, respectively, for all gel samples. The Stern–Volmer equation combined with moving boundary and Li-Tanaka models were used to explain the behavior of I_em during drying and swelling processes respectively. It is found that the desorption coefficient D_d decreased as κC contents were increased for a given temperature during drying. However, the cooperative diffusion coefficient, D_s presented exactly the opposite case. Conventional gravimetrical and volumetric experiments were also carried out during drying and swelling of PAAm–κC composite gels. It was observed that D_d and D_s values measured with the fluorescence method were found to be much larger than they were measured with the conventional methods.     

The State of Water in Human and Dog Red Cell Membranes

The apparent activation energy for the water diffusion permeability coefficient, P_d, across the red cell membrane has been found to be 4.9 ± 0.3 kcal/mole in the dog and 6.0 ± 0.2 kcal/mole in the human being over the temperature range, 7° to 37°C. The apparent activation energy for the hydraulic conductivity, L_p, in dog red cells has been found to be 3.7 ± 0.4 kcal/mole and in human red cells, 3.3 ± 0.4 kcal/mole over the same temperature range. The product of L_p and the bulk viscosity of water, η, was independent of temperature for both dog and man which indicates that the geometry of the red cell membrane is not temperature-sensitive over our experimental temperature range in either species. In the case of the dog, the apparent activation energy for diffusion is the same as that for self-diffusion of water, 4.6–4.8 kcal/mole, which indicates that the process of water diffusion across the dog red cell membrane is the same as that in free solution. The slightly, but significantly, higher activation energy for water diffusion in human red cells is consonant with water-membrane interaction in the narrower equivalent pores characteristic of these cells. The observation that the apparent activation energy for hydraulic conductivity is less than that for water diffusion across the red cell membrane is characteristic of viscous flow and suggests that the flow of water across the membranes of these red cells under an osmotic pressure gradient is a viscous process.     

Cell Volume Regulation in Cultured Human Retinal Müller Cells Is Associated with Changes in Transmembrane Potential

Müller cells are mainly involved in controlling extracellular homeostasis in the retina, where intense neural activity alters ion concentrations and osmotic gradients, thus favoring cell swelling. This increase in cell volume is followed by a regulatory volume decrease response (RVD), which is known to be partially mediated by the activation of K⁺ and anion channels. However, the precise mechanisms underlying osmotic swelling and subsequent cell volume regulation in Müller cells have been evaluated by only a few studies. Although the activation of ion channels during the RVD response may alter transmembrane potential (V_m), no studies have actually addressed this issue in Müller cells. The aim of the present work is to evaluate RVD using a retinal Müller cell line (MIO-M1) under different extracellular ionic conditions, and to study a possible association between RVD and changes in V_m. Cell volume and V_m changes were evaluated using fluorescent probe techniques and a mathematical model. Results show that cell swelling and subsequent RVD were accompanied by V_m depolarization followed by repolarization. This response depended on the composition of extracellular media. Cells exposed to a hypoosmotic solution with reduced ionic strength underwent maximum RVD and had a larger repolarization. Both of these responses were reduced by K⁺ or Cl⁻ channel blockers. In contrast, cells facing a hypoosmotic solution with the same ionic strength as the isoosmotic solution showed a lower RVD and a smaller repolarization and were not affected by blockers. Together, experimental and simulated data led us to propose that the efficiency of the RVD process in Müller glia depends not only on the activation of ion channels, but is also strongly modulated by concurrent changes in the membrane potential. The relationship between ionic fluxes, changes in ion permeabilities and ion concentrations –all leading to changes in V_m– define the success of RVD.     

FURTHER STUDIES ON THE KINETICS OF OSMOSIS IN LIVING CELLS

Using unfertilized eggs of Arbacia punctulata as natural osmometers an attempt has been made to account for the course of swelling and shrinking of these cells in anisotonic solutions by means of the laws governing osmosis and diffusion. The method employed has been to compute permeability of the cell to water, as measured by the rate of volume change per unit of cell surface per unit of osmotic pressure outstanding between the cell and its medium. Permeability to water as here defined and as somewhat differently defined by Northrop is approximately constant during swelling and shrinking, at least for the first several minutes of these processes. Permeability is found to be independent of the osmotic pressure of the solution in which cells are swelling. Water is found to leave cells more readily than it enters, that is, permeability is greater during exosmosis than during endosmosis.     

Osmotic Behavior of Oat Coleoptile Tissue in Relation to Growth

Efforts were made to estimate the water potential difference that is required, between rapidly growing oat coleoptile cylinders and dilute medium, to support the rate of water uptake involved in elongation, (a) by the traditional method of determining the concentration of mannitol in which the tissue neither gains nor loses water, and (b) by measuring the rates of osmotic exchanges induced by treating the tissue with different hypotonic mannitol concentrations. Both methods indicated large water potential differences (3 to 10 atm), in some cases approaching the osmotic pressure of the cells. However, indication was obtained that the rates of osmotic exchanges induced by mannitol solutions, and presumably also the equilibrium response sought in (a), are governed by the rate of diffusional exchange of mannitol with the free space rather than by the permeability of the tissue to water. Osmotic swelling of the tissue measured by immersing it in water after its turgor pressure had been reduced by evaporation, was at least two to four times more rapid than when mannitol was involved. The permeability to water estimated by the evaporation-immersion method indicated that rapidly elongating cylinders have water potentials between -0.8 and -2.5 atm, or between 10 and 25 per cent of their osmotic pressure.     

THE EFFECT OF SALT CONCENTRATION OF THE MEDIUM ON THE RATE OF OSMOSIS OF WATER THROUGH THE MEMBRANE OF LIVING CELLS

1. Using the unfertilized egg of the sea urchin, Arbacia, as osmometer, it was found that the rate with which water enters or leaves the cell depends on the osmotic pressure of the medium: the velocity constant of the diffusion process is higher when the cell is in concentrated sea water, and lower when the sea water medium is diluted with distilled water. Differences of more than tenfold in the value of the velocity constant were obtained in this way. When velocity constants are plotted against concentration of medium, a sigmoid curve is obtained. 2. These results are believed to indicate that cells are more permeable to water when the osmotic pressure of the medium is high than when it is low. This relation would be accounted for if water should diffuse through pores in a partially hydrated gel, constituting the cell membrane. In a medium of high osmotic pressure, the gel is conceived to give up water, to shrink, and therefore to allow widening of its pores with more ready diffusion of water through them. Conversely, in solutions of lower osmotic pressure, the gel would take up water and its pores become narrow.     

The entrance of water into beef and dog red cells

The rate constants for diffusion of THO across the red cell membrane of beef and dog, and the rate of entrance of water into the erythrocytes of these species under an osmotic pressure gradient have been measured. For water entrance into the erythrocyte by diffusion the rate constants are 0.10 ± 0.02 msec.^-1 (beef) and 0.14 ± 0.03 msec.^-1 (dog); the permeability coefficients for water entrance under a pressure gradient of 1 osmol./cm³ are 0.28 See PDF for Equation These values permit the calculation of an equivalent pore radius for the erythrocyte membrane of 4.1 A for beef and 7.4 A for dog. In the beef red cell the change in THO diffusion due to osmotically produced cell volume shifts has been studied. The resistance to THO diffusion increases as the cell volume increases. At the maximum volume, (1.06 times normal), THO diffusion is decreased to 0.84 times the normal rate. This change in diffusion is attributed to swelling of the cellular membrane.     

THE KINETICS OF EXOSMOSIS OF WATER FROM LIVING CELLS

1. The rate of exosmosis of water was studied in unfertilized Arbacia eggs, in order to bring out possible differences between the kinetics of exosmosis and endosmosis. 2. Exosmosis, like endosmosis, is found to follow the equation See PDF for Equation, in which a is the total volume of water that will leave the cell before osmotic equilibrium is attained, x is the volume that has already left the cell at time t, and k is the velocity constant. 3. The velocity constants of the two processes are equal, provided the salt concentration of the medium is the same. 4. The temperature characteristic of exosmosis, as of endomosis, is high. 5. It is concluded that the kinetics of exosmosis and endosmosis of water in these cells are identical, the only difference in the processes being in the direction of the driving force of osmotic pressure.