Relationship between lipid fluidity and water permeability of bovine tracheal epithelial cell apical membranes

Apical membrane vesicles were prepared from bovine tracheal epithelial cells. These membranes were enriched in alkaline phosphatase specific activity 35-fold compared to cellular homogenates. Steady-state fluorescence polarization studies of these membranes, using three fluorophores, demonstrated that they possessed a relatively low fluidity. Studies using the probe 1,6-diphenyl-1,3,5-hexatriene detected thermotropic transitions at 25.7 +/- 0.4 and 26.8 +/- 0.6 degrees C in these membranes and their liposomes, respectively. Analysis of the composition of these membranes revealed a fatty acyl saturation index of 0.59 +/- 0.02, a protein/lipid ratio (w/w) of 0.60 +/- 0.06, a cholesterol/phospholipid ratio (mol/mol) of 0.83 +/- 0.11, and a sphingomyelin/lecithin ratio (mol/mol) of 0.64 +/- 0.10. Membrane vesicles were osmotically active when studied by a stopped-flow nephelometric technique. Arrhenius plots of rates of osmotic water efflux demonstrated break points at approximately 28 and 18 degrees C, with activation energies of 16.7 +/- 0.2 kcal mol-1 from 35 to 28 degrees C, 8.3 +/- 0.5 kcal mol-1 from 28 to 18 degrees C, and approximately 3.0 kcal mol-1 below 18 degrees C. Treatment of membrane vesicles with benzyl alcohol, a known fluidizer, decreased lipid order (increased fluidity) and increased the rate of osmotic water efflux. The present results suggest that water crosses tracheal epithelial cell apical membranes by solubility-diffusion across the lipid domain and that increases in fluidity correlate with increases in the water permeability of these membranes.     

Alcohol and respiratory and body temperature changes during tepid water immersion

Resting subjects were immersed for 30 min in water at 22 and 30 degrees C after drinking alcohol. Total ventilation, end-tidal PCO2, rectal temperature, aural temperature, mean skin temperature, heart rate, and oxygen consumption were recorded during the experiments. Blood samples taken before the immersion period were analyzed by gas-liquid chromatography. The mean blood alcohol levels were 82.50 +/- 9.93 mg.(100 ml)-1 and 100.6 +/- 12.64 mg (100 ml)-1 for the immersions at 22 and 30 degrees C, respectively. There was no significant change in body temperature measured aurally or rectally, mean surface skin temperature, or heart rate at either water temperature tested. Total expired ventilation was significantly attenuated for the last 15 min of the immersion at 22 degrees C, after alcohol consumption as compared to the ventilation change in water at 22 degrees C without ethanol. This response was not consistently significantly altered during immersion in water at 30 degrees C. It is evident that during a 30-min immersion in tepid water with a high blood alcohol level, body heat loss is not affected but some changes in ventilation do occur.     

Thawing dog spermatozoa in just-boiled water: submersion time and effect on sperm quality compared to thawing in water at 70 degrees C

Dog spermatozoa have better quality after thawing in water at 70-75 degrees C instead of 35-38 degrees C. The aim of Experiment 1 was to determine the time needed to thaw 0.5 mL straws in just-boiled (98 degrees C) water and that of Experiment 2 to determine whether thawing frozen dog spermatozoa in just-boiled water will result in better quality than thawing in water at 70 degrees C. Prior to freezing the straws of Experiment 1, a Type J thermocouple with wire diameters of 0.08 mm (Osiris Technical Systems, Centurion, South Africa) was placed in the center of each of ninety-three 0.5 mL straws (IMV Technologies, L'Aigle, France) filled with extender (Biladyl* with 0.5%, v/v of Equex STM paste**) and 54 filled with extender plus 200 x 10(6)spermatozoa/mL (Minitüb, Germany (*) and Nova Chemical Sales, MA (**)). Thirty straws with extender were thawed in water at 70 degrees C and the others in just-boiled water. Temperatures inside straws were recorded 10 times/s during warming. Two ejaculates were then collected from each of eight dogs and one from each of three others. Extended ejaculates from the same dog were pooled, frozen 8 cm above liquid nitrogen, and 2 straws from each of the 11 batches thawed in water at 70 degrees C for 8s and 2 in just-boiled water for 6.5s. Sperm morphology and viability were assessed on eosin-nigrosin smears made after thawing and the percentage progressively motile spermatozoa was estimated immediately, 1, 2 and 3h after thawing. The optimal submersion time in just-boiled water was 6.5s for both sperm concentrations, resulting in average temperatures of 23.6+/-1.5 degrees C (+/-S.E.M.) and 24.9+/-1.6 degrees C inside straws with extender or extender plus spermatozoa (P=0.6). The temperature inside straws thawed in water at 70 degrees C was 13.6+/-1.7 degrees C after 8s. Apart from a 1.5% higher (P<0.05) mean percentage motile sperm 2h after thawing, thawing dog spermatozoa in just-boiled (98 degrees C) water holds no benefit over thawing in water at 70 degrees C, which is easier to do.     

Effect of high temperature and water stress on in vitro germination and growth in isolates of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin

In non-irrigated agricultural fields in tropical zones, high temperature and water stress prevail during the main cropping season. Natural epizootics of Beauveria bassiana on lepidopteran pests occur during winter. Application of B. bassiana during hot months when pest populations are at their climax may prove an effective management strategy. Therefore, 29 isolates of B. bassiana were tested for their ability to germinate and grow in temperature and water availability conditions prevailing during the pest season in these fields. The effect of temperature cycles with 8 h duration of high temperature fluctuating with 16 h duration of lower temperature (similar to field conditions); low water availability; and a combination of these two stress conditions was studied. Germination and growth assays were done at fluctuating temperature cycles of 32, 35, 38, and 42+/-1 degrees C (8 h)/25+/-1 degrees C (16 h) and in media with water stress created by 10, 20, 30, and 40% polyethylene glycol (PEG 6000). Assays set at a continuous temperature of 25+/-1 degrees C with no PEG in the medium served as controls. Stress was assessed as percentage germination or as growth relative to control. Isolates showing 90% growth relative to the control at temperature cycles including high temperatures of 35 and 38+/-1 degrees C were identified. One isolate (ARSEF 2860) had a thermal threshold above 43 degrees C. At 25 degrees C, all but one isolate of B. bassiana showed >90% growth relative to the control in 10% PEG (-0.45 MPa). Some isolates were found with >90% growth relative to control in medium having 30% PEG with water availability (1.33 MPa), nearly equivalent to that in soils which induce permanent wilting point of plants. When isolates that showed >90% growth relative to the control at both stress conditions, were stressed simultaneously, a decrease in growth was observed. Growth was reduced by approximately 20% at 35+/-1 degrees C (8 h)/25+/-1 degrees C (16 h) and 10% PEG and was affected to a greater degree in combinations of harsher stress conditions. The isolate ARSEF 2860 with a thermal threshold of >43 degrees C showed approximately 80% relative growth at a combined stress of 38+/-1 degrees C (8 h)/25+/-1 degrees C (16 h) and 10% PEG. These findings will aid the selection of isolates for use in field trials in hot or dry agricultural climates.     

The kinetics of water exchange across the chloroplast membrane

The kinetics of water exchange across the membrane of class II chloroplasts has been studied by two NMR methods. Both methods utilize Dy(en)3+ (en = ethylenediamine) to induce a transmembranal chemical shift the order of 40 Hz in the water proton resonance. The shift reagent is impermeant to the chloroplast membrane, inert as a redox reagent, soluble at millimolar concentrations at neutral pH, and associated with a large, virtually temperature independent molar shift (0.10-0.12 ppm/mM). Water exchange across the membrane is monitored by two independent experiments. In the first, chemical exchange causes line broadening in the water proton resonance in the high-resolution spectrum. Measurement of the incremental linewidth as a function of transmembranal chemical shift determines the exchange kinetics as well as the fractions of water protons in internal and external media. In the second experiment, chemical exchange causes the transverse relaxation time, as measured by the Carr-Purcell-Gill-Meiboom technique, to be dependent on the 180 degree pulse spacing. The two experiments, while independent of each other, depend on the same set of theoretical parameters. These parameters are overdetermined by simultaneous analysis of both experiments. The mean lifetime of a water proton in the inner thylakoid space is found to be 1.1 +/- 0.8 ms at 25 degrees C and 2.75 +/- 0.4 ms at 3 degrees C in NH2OH/EDTA-treated chloroplasts. Values derived from dark-adapted chloroplasts that are active with respect to oxygen evolution are 1.1 +/- 0.3 ms (25 degrees C) and 1.75 +/- 0.4 ms (3 degrees C). The internal thylakoid volume is also determined in principle by the data, but uncertainties in the membrane volume and the transmembranal chemical shift severely limits the accuracy of this measurement.     

Effect of water temperature on cooling efficiency during hyperthermia in humans.

We evaluated the cooling rate of hyperthermic subjects, as measured by rectal temperature (T(re)), during immersion in a range of water temperatures. On 4 separate days, seven subjects (4 men, 3 women) exercised at 65% maximal oxygen consumption at an ambient temperature of 39 degrees C until T(re) increased to 40 degrees C (45.4 +/- 4.1 min). After exercise, the subjects were immersed in a circulated water bath controlled at 2, 8, 14, or 20 degrees C until T(re) returned to 37.5 degrees C. No difference in cooling rate was observed between the immersions at 8, 14, and 20 degrees C despite the differences in the skin surface-to-water temperature gradient, possibly because of the presence of shivering at 8 and 14 degrees C. Compared with the other conditions, however, the rate of cooling (0.35 +/- 0.14 degrees C/min) was significantly greater during the 2 degrees C water immersion, in which shivering was seldom observed. This rate was almost twice as much as the other conditions (P < 0.05). Our results suggest that 2 degrees C water is the most effective immersion treatment for exercise-induced hyperthermia.     

Rehydration after exercise with fresh young coconut water,carbohydrate-electrolyte beverage and plain water

This is to cross-over study to assess the effectiveness of fresh young coconut water (CW), and carbohydrate-electrolyte beverage (CEB) compared with plain water (PW) for whole body rehydration and blood volume (BV) restoration during a 2 h rehydration period following exercise-induced dehydration. Eight healthy male volunteers (mean age and VO2max of 22.4 +/- 3.3 years and 45.8 +/- 1.5 ml min kg-1 respectively) exercised at 60% of VO2max in the heat (31.1 +/- 0.03 degrees C, 51.4 +/- 0.1% rh) until 2.78 +/- 0.06% (1.6 +/- 0.1 kg) of their body weight (BW) was lost. After exercise, the subjects sat for 2 h in a thermoneutral environment (22.5 +/- 0.1 degrees C; 67.0 +/- 1.0% rh) and drank a volume of PW, CW and CEB on different occasions representing 120% of the fluid loss. A blood and urine sample, and the body weight of each subject was taken before and after exercise and at 30 min intervals throughout a rehydration period. Each subject remained fasted throughout rehydration. Each fluid was consumed in three portions in separate trials representing 50% (781 +/- 47 ml), 40% (625 +/- 33 ml) and 30% (469 +/- 28 ml) of the 120% fluid loss at 0, 30 and 60 min of the 2 h rehydration period, respectively. The drinks given were randomised. In all the trials the subjects were somewhat hypohydrated (range 0.08-0.18 kg BW below euhydrated BW; p > 0.05) after a 2 h rehydration period since additional water and BW were lost as a result of urine formation, respiration, sweat and metabolism. The percent of body weight loss that was regained (used as index of percent rehydration) during CW, PW, and CEB trials was 75 +/- 5%, 73 +/- 5% and 80 +/- 4% respectively, but was not statistically different between trials. The rehydration index, which provided an indication of how much of what was actually ingested was used for body weight restoration, was again not different statistically between trials (1.56 +/- 0.14, 1.36 +/- 0.13 and 1.71 +/- 0.21 for CW, CEB and PW respectively). Although BV restoration was better with CW, it was not statistically different from CEB and PW. Cumulative urine output was similar in all trials. There were no difference at any time in serum Na+ and Cl-, serum osmolality, and net fluid balance between the three trials. Urine osmolality decreased after 1 h during the rehydration period and it was lowest in the PW trial. Plasma glucose concentrations were significantly higher compared with PW ingestion when CW and CEB were ingested during the rehydration period. CW was significantly sweeter, caused less nausea, fullness and no stomach upset and was also easier to consume in a larger amount compared with CEB and PW ingestion. In conclusion, ingestion of fresh young coconut water, a natural refreshing beverage, could be used for whole body rehydration after exercise.     

Diffusion of water in cat ventricular myocardium

The rates of diffusion of tritiated water (THO) and [14C]sucrose across cat right ventricular myocardium were studied at 23 degrees C in an Ussing-type diffusion cell, recording the time-course of increase in concentration of tracer in one chamber over 4--6 h after adding tracers to the other. Sucrose data were fitted with a model for a homogeneous sheet of uneven thickness in which the tissue is considered to be an array of parallel independent pathways (parallel pathway model) of varying length. The volume of the sucrose diffusion space, presumably a wholly extracellular pathway, was 23% of the tissue or 27.4 +/-1.7% (mean +/- SEM; n=11) of the tissue water. The effective intramyocardial sucrose diffusion coefficient, D8, was 1.51 +/- 0.19 X 10(-6)cm2.s-1 (n=11). Combining these data with earlier data, D8 was 22.6 +/- 1.1% (n=95) of the free diffusion coefficient in aqueous solution D degrees 8. The parallel pathway model and a dead-end pore model, which might have accounted for intracellular sequestration of water, gave estimates of DW/D degrees W (observed/free) of 15%. Because hindrance to water diffusion must be less than for sucrose (where D8/D degrees 8=22.6%), this showed the inadequacy of these models to account simultaneously for the diffusional resistance and the tissue water content. The third or cell-matrix model, a heterogeneous system of permeable cells arrayed in the extracellular matrix, allowed logical and geometrically reasonable interpretations of the steady-state data and implied estimates of DW in the cellular and extracellular fluid of approximately 25% of the aqueous diffusion coefficient.     

The equilibrium constants and enthalpy changes for the esterification of glycine by ethanol in water and ethanol- water mixtures

Equilibrium constants, defined on the basis of the moles of all reactants and products including water, were found to be 4.6, 1.0 and 0.8 for the esterification of the glycinc cation at 20 degrees in a dilute aqueous sytem, and systems having 0.5 : 1 and 1 : 1 molar ratios of ethanol to water, respectively. When corrections were made for deviations from ideality a value of 5 +/- 1 was obtained for all three systems. Enthalpy changes were determined calorimetrically for the dilute aqueous and equimolar ethanol-water systems, and in each case was close to 1 kcal mole(-). The entropy change was calculated as 6 cal mole(-1) deg(-1).DeltaG(0') for the hydrolysis of the glycine ethyl ester cation at 20 degrees and pH 7 was calculated as -7.5 kcal mole(-1).     

Voluntary dehydration and alliesthesia for water

The purpose of this experiment was to explore the complex relationship between fluid consumption and consumption factors (thirst, voluntary dehydration, water alliesthesia, palatability, work-rest cycle) during a simulated 14.5-km desert walk (treadmill, 1.34 m X s-1, 5% grade, 40 degrees C dry bulb/26 degrees C wet bulb, and wind speed of approximately 1.2 m X s-1). Twenty-nine subjects were tested (30 min X h-1, 6 h) on each of two nonconsecutive days. The subjects were randomly assigned to one of three groups: tap water (n = 8), iodine-treated tap water (n = 11), or iodine-treated flavored tap water (n = 10). The temperature of the water was 40 degrees C during one trial and 15 degrees C on the other. Mean sweat losses (6 h) varied between 1.4 kg (warm iodine-treated; 232 +/- 44 g X h-1) and 3.0 kg (cool iodine-treated flavored; 509 +/- 50 g X h-1). Warm drinks were consumed at a lower rate than cool drinks (negative and positive alliesthesia). This decreased consumption resulted in the highest percent body weight losses (2.8 and 3.2%). Cooling and flavoring effects on consumption were additive and increased the rate of intake by 120%. The apparent paradox between reduced consumption concomitant with severe dehydration and hyperthermia is attributed to negative alliesthesia for warm water rather than an apparent inadequacy of the thirst mechanism. The reluctance to drink warm iodine-treated water resulted in significant hyperthermia, dehydration, hypovolemia, and, in two cases, heat illness.     

Thermoregulation of weaned northern elephant seal (Mirounga angustirostris) pups in air and water

Fasting weaned northern elephant seal pups (Mirounga angustirostris) experience diverse environmental conditions on land and in water on a daily basis. Each environment undoubtedly induces distinct energetic costs that may vary for pups of differing body condition. To determine the energetic costs associated with different environmental conditions and whether costs vary between individuals, body mass, surface area, volume, body composition, resting metabolic rate, and core body temperature were determined for 17 weaned northern elephant seal pups from A?o Nuevo, California. Metabolic rate and body temperature were measured for pups resting in air (20.9 degrees +/-0.8 degrees C), cold water (3.8 degrees+/-0.4 degrees ;C), and warm water (14.5 degrees+/-0.2 degrees C). Resting metabolic rate increased with body mass (range: 62.0-108.0 kg) and was also correlated with lean mass and lipid mass. Metabolic rates ranged from 293.6 to 512.7 mL O(2) min(-1) and were lowest for pups resting in cold water. Thermal conductance, calculated from metabolic rate and core body temperature, ranged from 3.1 to 15.2 W degrees C(-1), with the highest values in air and the lowest values in cold water. Metabolic responses to the three environmental conditions did not differ with individual variation in body condition. For all elephant seal pups, a consequence of high lipid content is that thermoregulatory costs are greatest on land and lowest in cold water, a pattern that contrasts markedly with terrestrial mammals.     

Effect of water temperature on the immune response and infectivity pattern of white spot syndrome virus (WSSV) in freshwater crayfish

The susceptibility of two species of freshwater crayfish, Pacifastacus leniusculus and Astacus astacus, to white spot syndrome virus (WSSV) by intramuscular injection was compared and the results show that both species are susceptible to WSSV. The effect of water temperature on the development of white spot disease in crayfish was also studied. Crayfish were exposed to different temperatures after WSSV injection or oral exposure and the mortalities were recorded over a period of 45 days. No mortality was observed when crayfish were held at 4+/-2 degrees C or 12+/-2 degrees C and reached 100% when these crayfish transferred to 22+/-2 degrees C. The mortalities of nearly moribund crayfish at 22+/-2 degrees C with WSSV could be delayed after transfer to temperature below 16 degrees C. These results clearly show that low temperature affects the WSSV pathogenicity in crayfish. Moreover, haemocyte counts, phenoloxidase activity, mRNA levels of prophenoloxidase (proPO) and the lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) in crayfish exposed to various water temperatures were studied. Total haemocyte and granular cell counts of crayfish held at different temperatures were not significantly (P>0.05) different, except for the total haemocyte number at 18 degrees C was significantly (P<0.05) higher than in crayfish at 4 degrees C. The percentage of granular cells in crayfish held at 4 degrees C was the highest compared to crayfish maintained at other temperatures. The phenoloxidase activities in haemocyte lysate supernatant (HLS) of crayfish at all temperature groups remained similar. The amount of proPO-mRNAs in haemocytes was much higher than the amount of LGBP-m RNAs in all the experimental groups. However, there was no change in the level of pro PO-mRNA at the tested temperatures. Interestingly, the level of LGBP-mRNA of crayfish kept at 22 degrees C was much lower than in those held at lower temperatures. Proliferation of the haematopoietic tissues was higher at high temperatures which may support replication of WSSV, and explain the high mortality of crayfish with WSSV infection at high temperature. Based on these studies it is concluded that crayfish might act as a carrier of WSSV at low water temperature and could develop white spot disease if the water temperature is increased.     

Regional heat flows of resting and exercising men immersed in cool water

Trunk (HT), limb (HL), and whole-body (HDIR = HT + HL + Hforehead) skin-to-water heat flows were measured by heat flow transducers on nine men immersed head out in water at critical temperature (TCW = 30 +/- 2 degrees C) and below [overall water temperature (TW) range = 22-32 degrees C] after up to 3 h at rest and exercise. Body heat flow was also determined indirectly (HM) from metabolic rate corrected for changes in heat stores. At rest at TCW [O2 uptake (VO2) = 0.33 +/- 0.07 l/min, n = 7], HT = 52.3 +/- 14.2 (SD) W, HL = 56.4 +/- 14.6 W, HDIR = 120 +/- 27 W, and HM = 111 +/- 29 W (significantly different from HDIR). TW markedly affected HDIR but only slightly affected HM (n = 22 experiments at TW different from TCW plus 7 experiments at TCW). During light exercise (3 MET) at TCW (VO2 = 1.06 +/- 0.26 l/min, n = 9), HT = 122 +/- 43 W, HL = 130 +/- 27 W, HDIR = 285 +/- 69 W, and HM = 260 +/- 60 W. During severe exercise (7 MET) at TCW (VO2 = 2.27 +/- 0.50 l/min, n = 4), HT = 226 +/- 100 W, HL = 262 +/- 61 W, HDIR = 517 +/- 148 W, and HM = 496 +/- 98 W. Lowering TW at 7-MET exercise (n = 9, plus 4 at TCW) had no effect on HDIR and HM. In conclusion, resting HL and HT are equal. At TW less than TCW at rest, HDIR greater than HM, showing that unexpectedly the shell was still cooling. During exercise, HL increases more than HT but less than expected from the heat production of the working limbs. Therefore some heat produced by the limbs is probably transported by blood to the trunk. During heavy exercise, HDIR is constant at all considered TW; apparently it is regulated by some thermally dependent mechanism, such as a progressive cutaneous vasodilation occurring as TW increases.     

Early fluid and protein shifts in men during water immersion

High precision blood and plasma densitometry was used to measure transvascular fluid shifts during water immersion to the neck. Six men (28-49 years) undertook 30 min of standing immersion in water at 35.0 +/- 0.2 degrees C; immersion was preceded by 30 min control standing in air at 28 +/- 1 degrees C. Blood was sampled from an antecubital catheter for determination of blood density (BD), plasma density (PD), haematocrit (Ht), total plasma protein concentration (PPC), and plasma albumin concentration (PAC). Compared to control, significant decreases (p less than 0.01) in all these measures were observed after 20 min immersion. At 30 min, plasma volume had increased by 11.0 +/- 2.8%; the average density of the fluid shifted from extravascular fluid into the vascular compartment was 1006.3 g.l-1; albumin moved with the fluid and its albumin concentration was about one-third of the plasma protein concentration during early immersion. These calculations are based on the assumption that the F-cell ratio remained unchanged. No changes in erythrocyte water content during immersion were found. Thus, immersion-induced haemodilution is probably accompanied by protein (mainly albumin) augmentation which accompanies the intravascular fluid shift.     

Cardiorespiratory response to low-intensity walking in water and on land in elderly women.

The purpose of the present study was to determine whether or not the exercise intensity of water-walking for elderly women could be accurately prescribed by heart rate data obtained during treadmill exercise on land. Six healthy female volunteers, with a mean age of 62.2 +/- 4.2 years, took part in this study. Walking on land was performed on a treadmill. Each subject completed three consecutive 4-minute walks at a progressively increasing velocity (40, 60 and 80 m.min-1), with a 1-minute rest after both the first and second walks. The room temperature and relative humidity were 24.5 +/- 0.2 degrees C and 54.8 +/- 4.0%, respectively. Walking in water was performed in a Flowmill, which is a treadmill positioned at the base of a water flume. Each subject completed three consecutive 4-minute walks at a progressively increasing belt and water-flow velocity (20, 30 and 40 m.min-1), with a 1-minute rest after both the first and second walks. The water depth was at the level of the xiphoid process of each subject. The water temperature was 30.7 +/- 0.1 degrees C. The exercise intensity at the highest workrate was equivalent to 44.2 +/- 10.3% of the heart rate reserve (HRR) during water-walking and 38.4 +/- 4.7% of the HRR during land-walking. There was a highly significant linear relationship between heart rate (HR) and oxygen uptake (VO2) during both water-walking and land-walking. The relationship between HR and VO2 in both exercise modes was similar. Thus, the relationship of HR to VO2 derived from a treadmill-graded walking test on land may be used to prescribe exercise intensity for water-walking in thermoneutral water.     

Osmotic water permeability and regulatory volume decrease of rat thymocytes

Rat thymocytes displayed robust regulatory volume decrease (RVD) when suspended in NaCl-based hypotonic Ringer solutions. The RVD of thymocytes was completely abolished upon replacement of external Na+ ions with K+, indicating a role of coupled efflux of K+ and Cl- ions as a driving force of regulatory volume decrease. Osmotic water permeability (Pf) measured in KCl-based hypotonic solutions was (1.3 +/- 1.0 x 10(-4) cm/s at 25 degrees C and was temperature-dependent with low activation energy (Ea = 4.65 +/- 0.77 kcal/mol) characteristic to water transport through pores. HgCl2 and a sulfhydryl-blocking reagent, methyl methanethiosulphonate (MMTS), modulated the water permeability of thymocytes in a biphasic manner: inhibited at low dose (0.1-1 micromol/l) and restored or even enhanced at higher (10-100 micromol/l) concentrations. RVD paralleled the Pf: it was greatly suppressed at low dose of MMTS (sufficient to attenuate the water transport), but recovered at higher dose, when the water movement was restored. Therefore we suggest that thymocytes require the effective water transport for functional regulatory volume decrease.     

Effect of atrial natriuretic factor on the water permeability of endothelial cells.

Atrial natriuretic factor increases the water permeability of the whole endothelium. This study investigates how it would affect the transcellular osmotic water permeability of bovine artery endothelial cells. The cyclic-GMP production by the isolated cells was maximal for 10(-6)M atrial natriuretic factor within 30 minutes at 37 degrees C. The cyclic-GMP protein kinase cell concentration was 1.87 +/- 0.15 ng/mg protein. The control apparent water permeability of the cells measured by light scattering was 195 +/- 11 microns/sec (n = 5). Membrane folding revealed by light and scanning electron microscopy indicated that their true water permeability values would be close to 20-40 microns/sec, similar to the values for lipid membranes. The energy activation calculated from the temperature dependence of water permeability between 15 degrees C and 37 degrees C was 9.3 kcal/mol. This value suggests water movement through the lipid bilayer and not through water channels. Atrial natriuretic factor 10(-6)M did not significantly increase the water permeability of the cells. Hence, atrial natriuretic factor-stimulated increase in water permeability of the endothelium is more related to changes in paracellular water pathways than in transcellular water flux.     

Osmotic gradient-induced water permeation across the sarcolemma of rabbit ventricular myocytes

The mechanism of water permeation across the sarcolemma was characterized by examining the kinetics and temperature dependence of osmotic swelling and shrinkage of rabbit ventricular myocytes. The magnitude of swelling and the kinetics of swelling and shrinkage were temperature dependent, but the magnitude of shrinkage was very similar at 6 degrees, 22 degrees, and 37 degrees C. Membrane hydraulic conductivity, Lp, was approximately 1.2 x 10(-10) liter.N-1.s-1 at 22 degrees C, corresponding to an osmotic permeability coefficient, Pf, of 16 microns.s-1, and was independent of the direction of water flux, the magnitude of the imposed osmotic gradient (35-165 mosm/liter), and the initial cell volume. This value of Lp represents an upper limit because the membrane was assumed to be a smooth surface. Based on capacitive membrane area, Lp was 0.7 to 0.9 x 10(-10) liter.N-1.s-1. Nevertheless, estimates of Lp in ventricle are 15 to 25 times lower than those in human erythrocytes and are in the range of values reported for protein- free lipid bilayers and biological membranes without functioning water channels (aquaporin). Evaluation of the effect of unstirred layers showed that in the worst case they decrease Lp by < or = 2.3%. Analysis of the temperature dependence of Lp indicated that its apparent Arrhenius activation energy, Ea', was 11.7 +/- 0.9 kcal/mol between 6 degrees and 22 degrees C and 9.2 +/- 0.9 kcal/mol between 22 degrees and 37 degrees C. These values are significantly greater than that typically found for water flow through water-filled pores, approximately 4 kcal/mol, and are in the range reported for artificial and natural membranes without functioning water channels. Taken together, these data strongly argue that the vast majority of osmotic water flux in ventricular myocytes penetrates the lipid bilayer itself rather than passing through water-filled pores.     

不同含水量和温度下贮存中球孢白僵菌分生孢子活力与内贮营养的衰变

将球孢白僵菌(Beauveriabassiana)BBSG8702的未干燥孢子粉(含水量58.9±1.6%)和真空冷冻干燥孢子粉(含水量7.4±0.9%)置于4℃和20℃下贮存1个月,每隔5d取样测定活孢率和孢子内贮总糖和蛋白含量,发现含水量和贮存温度交互影响孢子的活力以及内贮总糖和蛋白质的代谢水平,各组合中的活孢率一般与内贮总糖和蛋白质代谢水平均存在显著或极显著相关性.在1个月的贮存期间,4℃下冻干粉总糖含量下降13.4%,蛋白质含量下降39.2%,清水中的萌发率下降32.0%,营养液中的萌发率仅下降6.7%,而未干燥孢子粉的相同指标分别下降42.4%、66.3%、96.4%和99%;在20℃下,冻干粉的上述指标分别下降了14.1%、38.2%、55.8%和 10.4%,而未干燥孢子粉则分别下降了 43.2%、65.4%、99.4%和98.4%. 显然,含水量影响活孢率和内贮营养衰变的幅度,而温度影响衰变的速度,但内贮营养的耗尽并不立即引起孢子失活,在供给外源营养之后孢子仍能萌发.将含水量降至4.0±0.9%的冻干粉贮存1年,4℃下活孢率由初始的99.0%下降至90.2%,而20℃下贮存的前165d活孢率下降较为缓慢,但此后急剧下降,至第240d时几乎全部失活.模拟分析表明,低含水量冻干粉在4℃和20℃下贮存的半衰期(即活孢率减少一半所需的时间)分别为1006d和197d.这些结果说明,白僵菌纯孢粉的含水量     

Permeability of human granulocytes to water: rectification of osmotic flow

The permeability of human granulocytes to water was studied for samples of that population with an electronic particle counter. Changes in volume were monitored as these samples were introduced suddenly into hypo- and hypertonic osmotica. Temperature and concentration sensitivity analyses of the permeability coefficients were carried out. An apparent rectification of water flow seems to occur with the water permeability being three times higher for endosmotic flow than for exosmotic flow. The estimated water permeability at the reference temperature 20 degrees C was for exosmotic flow 2.04 +/- 0.02 x 10(-12) kgmol N-1 s-1 with an apparent transfer energy of 4.76 +/- 0.09 x 10(7) J kgmol-1, and for endosmotic flow 6.05 +/- 0.07 x 10(-12) kgmol N-1 s-1 with an apparent transfer energy of 4.9 +/- 0.1 x 10(7) J kgmol-1.