Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. V—Rabbit (Oryctolagus Cuniculus)

• 1.1. The diffusional water permeability (P_d) of rabbit red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulfonate (PCMBS).
• 2.2. The values of P_d were around 6.3 × 10⁻³ cm/sec at 15°C, 7.0 × 10⁻³cm/sec at 20°C, 8.0 × 10⁻³ cm/sec at 25°C, 9.1 × 10⁻³ cm/sec at 30°C and10.7 × 10⁻³ cm/sec at 37°C.
• 3.3. Systematic studies on the effects of PCMBS on water diffusion indicated that the maximal inhibition was reached in 15 min at 37°C with 0.5 mM PCMBS.
• 4.4. The values of maximal inhibition were around 71–74% at all temperatures.
• 5.5. The basal permeability to water was estimated as 1.6 × 10⁻³cm/sec at 15°C, 2.0 × 10⁻³cm/sec at 20°C, 2.4 × 10⁻³cm/sec at 25°C, 2.6 × 10⁻³cm/sec at 30°C, and 3.1× 10^{−3 cm/secat 37°C}.
• 6.6. The activation energy of water diffusion was around 18 kJ/mol and increased to 27 kcal/mol after incubation with PCMBS in conditions of maximal inhibition of water diffusion.
• 7.7. The membrane polypeptide electrophoretic pattern of rabbit RBCs has been compared with its human counterpart.
• 8.8. The rabbit membrane contained a higher amount of spectrin (bands 1 and 2), while the band 6 (glyceraldehyde-3-phosphate dehydrogenase) was markedly less intense.
• 9.9. Considerable differences in the electrophoretic patterns of the two sources of RBC membranes appeared in the bands migrating in the band 4.5 region and in front of band 7, where some polypeptides were apparent in higher amounts in the rabbit RBC membrane.

     

Changes in high-energy phosphate metabolism in the water scorpion,Ranatra chinensis,under cold water-warm water stress

• 1.1. To evaluate changes in high-energy phosphate metabolism in the water scorpion (Ranatra chinensis) under restraint and cold water-warm water stresses, in vivo [³¹P]NMR spectra were obtained.
• 2.2. Under restraint stress, arginine phosphate (Arg-P) decreased by 10% after 1 hr and remained at that level thereafter, while β-ATP showed negligible changes over 6 hr.
• 3.3. As the water temperature gradually increased or decreased, the relative concentration of Arg-P decreased due to enzyme regulation.
• 4.4. Repeated cold water-warm water stress, which consisted of repeated 15 min exposures to cold water (5°C) followed by 15 min exposures to warm water (30°C) caused distinct decreases in Arg-P and β-ATP concentration. These decreases were dependent on the frequency of exposure.
• 5.5. Phosphomonoesters (PME) increased not only with restraint stress but also with cold water-warm water stress.
• 6.6. The effect of cold water-warm water stress on high-energy phosphate metabolism was greater than that of restraint stress.

     

γ-Glutamyltranspeptidase in echinoderm eggs and larvae: fertilization-dependent and developmentally-induced changes in specific activity

• 1.1. γ-Glutamyltranspeptidase is present in echinoderm eggs and larvae: in homogenates the level of activity is comparable to that of rat cerebral cortex.
• 2.2. In eggs of Lytechinus pictus, fertilization induces an early rapid and sustained (5 min–6 hr) 37% increase in the activity of γ-glutamyltranspeptidase in homogenate fractions.
• 3.3. Relative to these homogenate levels, the specific activity of γ-glutamyltranspeptidase are ≈60% lower in 40,000 g supernatant fractions and 2.7-fold higher in 40,000 g particulate fractions in both unfertilized and 15 min post-fertilized Lytechinus pictus eggs.
• 4.4. The subcellular distribution of γ-glutamyltranspeptidase is the same in both unfertilized and 15-min post-fertilized Lytechinus pictus eggs: 78% in 40,000 g particulate fractions, 22% in 40,000 g soluble fractions.
• 5.5. In both unfertilized and 15 min post-fertilized eggs of Lytechinus pictus the enzyme responds to heat (50 vs 37°C) by activation in a similar manner: 1.72- and 1.68-fold homogenates; 2.6- and 3.0-fold in supernatants; 1.97- and 1.90-fold in particulate fractions.
• 6.6. In homogenates of Pisaster ochraceous larvae, γ-glutamyltranspeptidase activity increases steadily during the course of larval development: relative to the low activity at day 5, activities exhibit an increase of 1.2-, 2.0-, 3.1- and 5.4-fold at days 10, 16, 22 and 28, respectively.

     

Behavioural thermoregulation and critical thermal limits of Macrobrachium acanthurus (Wiegman)

• (1)The preferred temperatures of Macrobrachium acanthurus were determined for prawns acclimated to 20°C, 23°C, 26°C, 29°C and 32°C, and the final preferendum estimate was (29.5°C).
• (2)The critical thermal minima (CTMin) and maxima (CTMax) were 11.0°C, 12.1°C, 13.0°C and 14.8°C, and 34.2°C, 35.0°C, 36.1°C and 39.8°C, respectively.
• (3)The zone of thermal tolerance assessed using the CTMin and CTMax boundaries was 644°C².
• (4)The acclimation response ratio was between 0.33 and 0.62.
• (5)To cultivate this species in the southeastern region of México it should be done in not <15°C (CTMin) during the winter and below 38°C in summer (CTMax).

     

The maltase of the house cricket Acheta domesticus L. (orthoptera: gryllidae)

• 1.1. The optimal pH of maltase of the house cricket was 5.6 and the optimal temperature was 40°C. The apparent activation energy was 5300 cal/mol
• 2.2. The 50% inactivation time for maltase at 50°C was 69 min and the 50% inhibition concentration for mercuric chloride was 163μmole.
• 3.3. Maltase was only produced by the caecal tissues but its highest activity was found in the foregut lumen and after 5 days of starvation, only 44.6% of the total maltase activity remained.

     

Metabolic and functional characteristics of erythrocytes from Glycera and Noetia

• 1.1. Annelid and molluscan red blood cells (RBC) may de differentiated metabolically from vertebrate RBC by their increased permeability to substrate, their magnitude of amino acid catabolism and their higher aerobic metabolism.
• 2.2. At 22°C, Glycera and Noetia RBC oxidize glucose and glutamate to CO₂ without accumulation of either d- or l-lactate. By comparison, the oxidation of glutamate by rat and chicken RBC is negligible at this temperature despite its incorporation into the cells.
• 3.3. At 37°C, chicken RBC oxidize glutamate at a rate 4 times greater than at 22°C, with oxygen uptake still lower than that in Noetia RBC at 32°C. At 37°C, rat RBC do not increase their oxidation of glutamate above that at 22°C, but oxygen uptake increases to slightly more than half that of chicken RBC.
• 4.4. Our finclings indicate that RBC of these two invertebrate species have both a higher aerobic metabolism and lower anerobic capacity than vertebrate RBC.
• 5.5. Moreover, the annelid and molluscan RBC have a relatively lower activity of the pentose phosphate (PPO₄) pathway than vertebrate RBC, as evidenced by their higher thermal sensitivity of oxygen uptake and their higher *C₁O₂/*C₆O₂ isotope ratio.

     

Temperature-dependence of spectroscopic and catalytic properties of the eel (Anguilla anguilla) liver mitochondrial F1-ATPase

• 1.1. F₁-ATPase from eel liver mitochondria at low concentrations preserves unaltered the enzymatic activity for more than 20 min over a temperature range of 6–36°C.
• 2.2. The Arrhenius plot of ATP hydrolysis at saturating substrate concentration appears biphasic with a break-point at 16°C and activation energies of 14.4 and 56.1 kJ/mol.
• 3.3. The ultraviolet, fluorescence and circular dichroism spectra of the enzyme, below and above 16°C, have been recorded; the fluorescence emission spectra of F₁-ATPase excited at 275 nm, and the circular dichroism spectra, are different at the two temperatures examined.
• 4.4. It is concluded that temperature induces two different conformational states of F₁-ATPase with different catalytic properties.
• 5.5. Ultraviolet spectroscopic features and temperature-dependence of eel liver mitochondrial F₁-ATPase are discussed in relation to mammalian F₁-ATPases.

     

A study of the effect of heat shock and metal ions on protein synthesis in Neurospora crassa cells

• 1.1. Neurospora cells were grown at 28°C for 14hr and subjected to heat shock (HS) at 48°C for 45 min. Protein synthesis profiles, monitored by labelling with [³⁵S]methionine and one and two-dimensional electrophoresis, revealed nine heat shock proteins (HSPs).
• 2.2. Crossed-immunoelectrophoresis revealed five polypeptides in the shocked cell extracts that were not detectable in normal cells.
• 3.3. Synthesis of HSPs occurred rapidly during the shock treatment and ceased upon transfer to normal conditions. One of the HSPs—~43 K in size—may be a developmentally-regulated protein.
• 4.4. Metal ions—cadmium, zinc, manganese, copper—did not elicit a stress response when used alone but appeared to modulate the heat shock response.

     

A thioredoxin from the extreme thermophilic Archaeon Sulfolobus solfataricus

• 1.1. A purification procedure for a thioredoxin from the extremophilic archaeon Sulfolobus solfataricus is described.
• 2.2. The thioredoxin is active in the dithiothreitol-dependent reduction of insulin disulfide bonds.
• 3.3. The thioredoxin is a monomer of 24,800 Da; it is an acidic protein with a pi of 4.5.
• 4.4. The protein is stable to heating for 3 hr at 90°C.
• 5.5. The amino acid composition of S. solfataricus thioredoxin is reported.

     

Influence of environmental factors on the deep and skin temperature in the European bison,Bison bonasus (L.)

• 1.1. In 43 European bison divided into three groups (Group A, 3–8-month-old calves; Group B, 18-month-7-year-old young bison; Group C, 12–24-year-old bison) the rectal, humerus region and abdomen region temperatures were measured.
• 2.2. The experiments were carried out in winter months, from mid-December to mid-March.
• 3.3. The mean rectal temperatures changed from 38.55°C in calves to 38.15°C in the oldest bison.
• 4.4. The mean temperatures of the humerus region changed from 20.69°C in calves to 21.49°C in older bison.
• 5.5. The mean temperatures of the abdomen region changed from 20.79°C in calves to 22.17°C in older bison (Gr. B).
• 6.6. The cluster analysis divided the bison into four groups named hot, warm, cool and cold bison.
• 7.7. Only air temperature measured 2 m above the ground and snow cover influenced the integrated bison temperature. Age, sex and mass as well as some environmental factors had no influence.
• 8.8. Measurements made 1 to nearly 4hr after a bison's death showed a drop in rectal temperature and mostly increases in temperatures of the humerus and abdomen regions.

     

The effect of temperature on the swimming of a teleost (Clupea harengus) and an ascidian larva (Dendrodoa grossularia)

• 1.1. Using a high-speed video system operating at 400 frames/sec, the effects of temperature on tail beat frequency, swimming speed and stride length were examined in newly hatched larvae of herring (Clupea harengus L.) and in tadpole larvae of the ascidian Dendrodoa grossularia van Beneden.
• 2.2. The effect of temperature was linear; the tail beat frequency of 8 mm-long herring larvae increased from 19 Hz at 5.6°C to 37 Hz at 14.9°C (Q₁₀ = 2.04); that of 2 mm-long Dendrodoa larvae increased from 10 Hz at 9.6°C to 23 Hz at 18.1°C (Q₁₀ = 2.52).
• 3.3. Burst swimming speeds of herring larvae increased from 80 mm/sec at 5°C to 150 mm/sec at 15°C, stride length remaining constant at about 0.5 of the body length for each tail beat.
• 4.4. More continuous swimming of Dendrodoa increased from 4.0 mm/sec at 10°C to 11.5 mm/sec at 18°C, the stride length increasing from about 0.15 to 0.25.

     

Sea-water tolerance in freshwater-resident arctic charr (Salvelinus alpinus)

• 1.1. Freshwater-resident Arctic charr acclimated for 2 months at 8°C, 15% were divided into four experimental groups in July and exposed to 1 and 8°C in 15 and 34% salinity.
• 2.2. Only slight changes in gill Na-K-ATPase activity, blood plasma osmolality and blood plasma concentrations of Cl⁻ and Mg²⁺ were found for the fish exposed to 1 or 8°C in brackish water.
• 3.3. When exposed to sea-water at 8°C, an increase in osmolality and in concentrations of Cl⁻ and Mg²⁺ took place during the first 2–3 days, after which it levelled off.
• 4.4. If exposed to sea-water at 1°C, however, marked increases were found for all parameters measured and all the fish were dead within 5 days of exposure.
• 5.5. These results show that freshwater-resident Arctic charr—if acclimated to brackish water—can survive in sea-water during summer if the environmental temperature is not too low.

     

Thermostability and activation by divalent cations of the membrane-bound inorganic pyrophosphatase of Rhodospirillum rubrum

• 1.1. The activation energy of the membrane bound H⁺-pyrophosphatase is 44.9 k J·mol⁻¹, for the detergent solubilized enzyme is 55.9 kJ·mol⁻¹.
• 2.2. The Arrhenius plots obtained for pyrophosphatases of Rhodospirillum rubrum show no breaks.
• 3.3. At 70°C, the membrane-bound pyrophosphatase is more stable in the presence of either Mg²⁺ or Zn²⁺ than in their absence.
• 4.4. At 65°C, an activator effect of Mg²⁺ or Zn²⁺ was observed. Nevertheless, at 70°C no activation was obtained.
• 5.5. The activator effects of Mg²⁺ or Zn²⁺ were depended of their concentration.

     

The regulation of glucose 6-phosphate dehydrogenase from Dicentrarchus labrax (bass) liver

• 1.1. Kinetic constant values of the reaction catalyzed by bass liver glucose 6-phosphate dehydrogenase show to be modified between 10 and 40°C.
• 2.2. The Arrhenius plot between 10 and 50°C shows two slopes with different activation energies.
• 3.3. These results suggest a regulation of this enzyme by environmental temperature.
• 4.4. Kinetics of ATP inhibition were examined between pH 6.2 and 7.8: patterns and K_i values obtained are affected by the pH variation.
• 5.5. NADH is an effective inhibitor of bass glucose 6-phosphate dehydrogenase but this enzyme does not show NAD-linked activity.
• 6.6. Kinetics of pyridoxal 5′-phosphate inhibition have indicated the presence of a lysine in the catalytic site for NADP⁺.

     

Temperature acclimation in respiratory and cytochrome c oxidase activity in common carp (Cyprinus carpio)

• 1.1. Common carp (Cyprinus carpio) exposed to experimental temperatures of 12, 18, 24, 30 or 36°C for a 4-week period were used to investigate the effect of temperature acclimation on the frequency of opercular movement (FOM), growth and cytochrome c oxidase (CCO) activity in heart, liver and muscle.
• 2.2. An exponential relationship between FOM and temperature after the first week (10₁₀ =1.76) disappeared after the second week.
• 3.3. The initially high FOM at temperatures of 30 or 36°C and the low FOM at 18 or 12°C changed over 4 weeks to approach the FOM of fish at 24°C.
• 4.4. This change in the relationship of FOM to temperature from highly dependent to independent appeared to be thermal compensation.
• 5.5. Heart and liver CCO activities were significantly affected by temperature, with the lowest activity at the approximate optimum temperature for growth, 24°C.
• 6.6. Highest CCO activities for heart and liver occurred at both the highest and lowest temperatures.
• 7.7. Among the three tissues, heart CCO activity was generally the highest and most affected by acclimation temperature.
• 8.8. Muscle tissue had the lowest CCO activity and was unaffected by temperature.
• 9.9. The high CCO activity at a cold acclimation of temperature 12°C was probably due to thermal compensation and the high activity at 36°C may have been a result of thermal stress.

     

Effect of temperature and salinity on the oxygen consumption of laboratory produced Penaeus californiensis postlarvae

• 1.1. The oxygen consumption by P. californiensis postlarvae (mean wt = 0.38 g) was determined at five different temperatures and four salinities.
• 2.2. The O₂ in each chamber was recorded at 10 min intervals for 1 hr. The time course of oxygen depletion was independent of O₂ concentration down to 1.6 mg/l.
• 3.3. Oxygen consumption increased with temperature from 0.0045 mg/g/min at 19°C, to 0.0142 mg/g/min at 35°C. The thermal coefficient (Q₁₀) indicated a very high sensitivity of the postlarvae to temperature variations at 19–23°C.
• 4.4. The results show that oxygen consumption significantly depends on temperature (P < 0.001) while salinity has only a marginal effect.

     

Purification and characterization of the amylolytic enzymes of Saccharomycopsis fibuligera

• 1.1. A complex of extracellular amylolytic enzymes produced by Saccharomycopsis fibuligera KZ, grown on fine fibre (waste product from corn starch production) and corn-steep liquor, has been studied.
• 2.2. α-Amylases and glucoamylases, as the main representatives of this complex, were separated by hydrophobic chromatography on Spheron 300 LC.
• 3.3. Individual isoenzymes of one type were separated on FPLC-Mono Q.
• 4.4. The relative molecular weight of α-amylases is 54,000, glucoamylases 62,000, maximal activity is reached by both enzymes between pH 5.0 and 6.2 at a temperature of 40–50°C.
• 5.5. Glucoamylases have a higher stability of the native structure than α-amylases, they retain 55% of their original activity, even after 10 min of incubation at 100°C.

     

Evidence that protein kinase C is involved in regulating glucose transport in the adipocyte

• 1.1. The role of protein kinase C in the mechanism of stimulation of glucose transport in rat adipocytes was investigated.
• 2.2. Glucose transprt was stimulated by dioleoylglycerol (DOG), tetradecanoyl phorbol acetate (TPA) and phospholipase C (PLC).
• 3.3. Agents that inhibit protein kinase C (polymyxin B, gossypol and quercitin) also inhibited glucose transport that had been stimulated by DOG, TPA, PLC and insulin.

     

Effects of temperature and metabolic inhibitors on contraction of anterior byssus retractor muscle of Mytilus

• 1.1. Anterior byssus retractor muscle of Mytilus (ABRM) was stimulated to contract by ACh (acetylcholine) and effects of temperature (5–30°C), FDNB (1-fluoro 2,4 dinitro-benzene) and IAA (iodoacetic acid) on tension response were examined.
• 2.2. Isometric tension was highest at the temperature range of 10–20°C and decreased at higher and lower temperature than that range.
• 3.3. The rate of tension decay after washing of ACh was accelerated by the increase of temperature.
• 4.4. Tension redevelopment after release of 1 % during contraction was much smaller at 5°C than at 20°C.
• 5.5. Tension development by ACh and the rate of tension decay after washing of ACh were remarkably decreased by the treatment of FDNB or IAA.
• 6.6. The above results were discussed from the viewpoint that energy metabolism might be related to catch.

     

Characterization of two distinct latent proteinases associated with myofibrils of crucian carp (Carassius auratus cuvieri)

:

• 1.1. Enzymatic properties of two distinct proteinases tightly associated with crucian carp myofibrils were characterized.
• 2.2. These proteinases were latent but activated at 50 and 60°C, respectively.
• 3.3. The optimum pH of 50°C-proteinase was neutral-alkaline, while that of 60°C-proteinase was weak acid-neutral pH.
• 4.4. Both proteinases required more than 1% NaCl for the activity, but 50°C-proteinase was partially inhibited at higher concentrations of NaCl.
• 5.5. Both proteinases were regarded as trypsin-like proteinases belonging to a serine proteinase family, but only 60°C-proteinase was sensitive to urea, n-butanol and iso-propanol.