Kinetic analysis of a cytoplasmic casein kinase II from Artemia sp.

• 1.1. A cytoplasmic casein kinase II (CKII) has been purified more than 10,000-fold from Artemia sp.
• 2.2. The reaction mechanism of the cytoplasmic CKII was determined to be random bi bi, using ATP and casein as substrates, which is in agreement with the results obtained for a DrosophilaCKII [Glover, Shelton and Brutlag (1983) J. biol. Chem.258, 3258–3265]. K_m values for ATP and casein are 8 μM and 0.2 mg/ml respectively. The binding of either substrate lowers the enzyme-affinity for the other by a factor α = 1.65.
• 3.3. In vitro, the enzyme is inhibited by poly(A)²-mRNA and 2,3-diphosphoglyceric acid (2,3-DPG). The inhibition by 2,3-DPG is due to competition with the protein substrate.
• 4.4. The possible in vivo effects of these inhibitors in CKII-mediated translational regulation is discussed.

     

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.

     

Respiration in the eastern water dragon,Physignathus lesueurii (agamidae)

• 1.1. Some aspects of the gas exchange system of a diving lizard, Physignathus lesuewii were studied.
• 2.2. Breathing patterns were analysed.
• 3.3. Breathing rate increases logarithmically with temperature and Q₁₀ = 1.8. LogBR = −0.237 + 0.0256 T.
• 4.4. Gas tensions in lung air and arterial and venous blood were measured. Arterial pH declines with increasing temperature.
• 5.5. Temperature has a marked effect on oxygen affinity of the blood (ΔH = −10.1 kcal mol⁻). A Bohr effect was also noted.
• 6.6. CO₂ equilibrium curves were drawn.
• 7.7. The results are considered with a view to anticipating the efficiency of the gas exchange system of this species under conditions of variable temperature and during diving.

     

Characterization of NaK ATPase from the gills of the freshwater prawn Macrobrachium rosenbergii (de Man)

• 1.1. The specific activity of Na-K ATPase was determined from the microsomal preparation of gills dissected from adult Macrobrachium rosenbergii.
• 2.2. Maximal ATPase activity was achieved at a substrate concentration of 0.5 mM ATP.
• 3.3. Optimal enzyme activity was obtained at pH of 7.5.
• 4.4. The Arrhenius plot of Na-K ATPase activity revealed a marked discontinuity at 30°C. “Mg” ATPase activity did not exhibit a marked discontinuity.
• 5.5. The E_a for Na-K ATPase and “Mg” ATPase was 14.6 kCal/mole and 9.31 kCal/mole respectively. Q₁₀ values for Na-K ATPase was 2.34 and for “Mg” ATPase 1.65.
• 6.6. ATPase activity and gill homogenate protein concentration exhibited a linear relationship up to 130 μg protein/ml.
• 7.7. Na-K ATPase activity was inhibited by 10⁻³ M ouabain. It was equally inhibited by the removal of K⁺ from the reaction medium.

     

Specific dynamic action (SDA) in the supralittoral isopod,Ligia pallasii: Relationship of growth to SDA

• 1.1. The relationship of Specific Dynamic Action (SDA) to growth was examined in the supralittoral isopod Ligia pallasii using a seaweed diet fed at different rations.
• 2.2. Animals increased in live weight by 33% on an ad libitum or 100% diet and by 2% on a 20% ration over a 10-week period.
• 3.3. Weight-specific VO₂ was significantly higher in animals eating the 100% diet than in ones eating the 20% diet. Decline in VO₂ with time in animals on the 20% diet was probably due to poor health associated with a maintenance ration.
• 4.4. SDA per unit weight of food eaten was 18% higher in the 20% diet group than in the 100% one, and values remained constant over time in both groups.
• 5.5. k₁ growth efficiencies (production/consumption) were higher in animals on 100% ration than in ones on 20% ration. Efficiencies declined with time in both diet-groups and fell below zero in the 20% ration-group, coincidental with weight-loss in some of the animals.
• 6.6. Overall SDAs for the 10-week period were positively correlated with growth (r₂ = 0.77), but there was no way to separate this from amounts eaten as an effect on SDA.

     

The influence of aerial exposure on gas exchange and cardiac activity in the shore crab,Carcinus maenas (L.)

• 1.1. The cardiovascular physiology of adult Carcinus maenas (L.) emerging into air has been investigated at three different air temperatures.
• 2.2. Transition from seawater to air or vice versa triggered transient increases in cardiac and locomotor activity.
• 3.3. However, crabs became inactive 5–10 min after emerging from seawater (15°C) into air at the same temperature (15°C) or at lower temperatures (12–13°C) and heart rate fell.
• 4.4. At higher air temperatures (18–20°C) heart rate rose but to a lesser extent than predicted from aquatic Q₁₀ heart-rate values.
• 5.5. Crabs were again quiescent in aerial conditions.
• 6.6. Mean arterial oxygen tension (Pa_o2) was ~ 74 mmHg in submerged crabs but fell to ~ 38 mmHg in air while mean arterial carbon dioxide tension (Pa_o2) increased from 1 to 4 mmHg resulting in respiratory acidosis.
• 7.7. A model of gill function is proposed to explain the development of internal hypoxia in air.
• 8.8. The results are discussed in relation to the distribution of adult and juvenile C. maenas in situ.

     

Length of hibernation and the brain's influence on temperature-induced spermatogenic DNA synthesis in Helix aspersa

• 1.1. In ovotestis of hibernating snails, the male cell line is very sensitive to temperature. An elevated temperature from 5 to 25°C induces spermatogenic DNA synthesis and formation of spermatids and spermatozoa in 4 weeks.
• 2.2. The DNA synthesis gradually increases in male cell line with the lengthening of hibernation.
• 3.3. Brain removal produces a marked increase of DNA synthesis after 1 and 6 months of hibernation (× 3.5 and 3.3) but not after 12 months.

     

Functional studies on the single component hemoglobin from an Amazon knife fish,Sternopygus macrurus

• 1. The whole blood of the non-air-breathing gymnotid teleost,Sternopygus macrurus, is half-saturated with oxygen at 5.2 mm Hg (apparent value) at 30°C in the absence of CO₂. Addition of 5.6% CO₂ causes the apparentP₅₀ value to increase over 3-fold.
• 2. The oxygen affinity of the stripped single-component hemoglobin at 20°C increases about 20-fold between pH 5.8 and 8.6 in the absence of ATP. This difference increases to 100-fold in the presence of 1 mM ATP.
• 3. A substantial Root effect is present: the stripped hemoglobin is only 70% saturated with O₂ at pH less than 6 when equilibrated with air.
• 4. The value of the Hill coefficient,n, is maximal near pH 7.0–7.5, and approaches 1.0 at high pH. The value is about 1.5 at low pH in the absence of ATP and 1.0 in the presence of 1 mM ATP.
• 5. The O₂ dissociation kinetics are heterogeneous at all pH values but most heterogeneous at low pH. The rate increases substantially as the pH decreases.
• 6. The CO combination kinetics as measured by the stopped flow technique are largely homogeneous except at high pH, but the CO combination kinetics after flash photolysis are markedly heterogeneous.

     

The suitability of using cryopreservation of spermatozoa for the conservation of genetic diversity in African catfish (Clarias gariepinus)

• 1.1. The effect of cryopreservation of semen on expected Hardy-Weinberg proportions in the f₁ progeny of selected breeding pairs was evaluated.
• 2.2. Four different African catfish breeding pairs were selected, each pair displaying different heterozygous alleles at the glucose-6-phosphate isomerase-1 or 2 loci.
• 3.3. Equal volumes of ova from each female were artificially inseminated with cryopreserved or fresh semen obtained from males possessing corresponding genotypes.
• 4.4. A comparison of growth rates between f₁ groups of offspring produced from fresh and cryopreserved semen was made.
• 5.5. The G-test for goodness of fit showed no significant differences from expected Hardy-Weinberg proportions for allele frequencies obtained in the f₁ progeny.
• 6.6. The application of cryopreservation for the conservation of genetic diversity is discussed.

     

Factors affecting oxygen consumption in wild-caught yellow-bellied marmots (Marmota flaviventris)

• 1.1. All age groups gained mass during the active season, but mass-gain of adult females was delayed during lactation.
• 2.2. The relationship of body mass to metabolic rate varied widely; when the relationship was significant, R² varied from 10.3 to 72.6%. Body mass affects V_O2 more during lactation than at any other period.
• 3.3. Mean VinO₂ of adult males was higher in June than that of adult, non-lactating females.
• 4.4. V_O2 of reproductive females was significantly higher during lactation than during gestation or postlactation because specific V_O2 varied. Specific V_O2 of non-reproductive females declined over the active season.
• 5.5. Specific V_O2 of all age groups declined between the premolt and postmolt periods. The reduced maintenance costs can contribute 20–46% to daily growth.
• 6.6. Observed V_O2 was lower than the value predicted from intraspecific or interspecific Bm:M regressions.
• 7.7. V_O2 of wild-caught marmots was lower than that of marmots maintained in the laboratory, probably because of dietary differences.
• 8.8. Because basal metabolism is a stage on a food-deprivation curve, we suggest that basal metabolic rate is not an appropriate measure of the metabolic activity of free-ranging animals.

     

Functional properties of hemoglobin and whole blood in an aquatic mammal,the Amazonian manatee (Trichechus inunguis)

• 1. Hematocrit (43%) and O₂ binding capacity (18.8 ml O₂/100 ml blood) ofTrichechus inunguis blood are low compared to the values for other diving mammals but are similar to those for land mammals.
• 2. Stripped manatee Hb is similar to human Hb A in its sensitivity to pH, 2,3-diphosphoglycerate and CO₂, but less sensitive to temperature and more prone to dissociate into dimers.
• 3. The unique Hill plots exhibit no cooperativity below 30% O₂-saturation indicating a highly stabilized T or low-affinity state(s); such asymmetric Hill plots together with biphasic O₂-binding kinetics could mean chain heterogeneity.
• 4. The pH dependence of oxygen binding by the apparent T state, hemoglobin as seen in the Hill plots, is enhanced by 2,3-diphosphoglycerate but eliminated by CO₂.

     

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.

     

Body temperature,oxygen consumption,evaporative water loss and heart rate in the fennec

ab]

• 1.1. Fennecs show marked diurnal variations of body temperature and heart rate.
• 2.2. Basal metabolic rate (0.358 ml/ghr) is 39% lower than predicted by body mass, minimal conductance is reduced for 23%.
• 3.3. Fennecs have a wide thermoneutral zone (23.4–32.0°C) and a low rate of evaporative water loss.
• 4.4. Basal heart rate is considerably reduced. Oxygen pulse increases with decreasing ambient temperature. The higher oxygen demands below thermal neutrality, however, are met primarily by a rise in heart rate.
• 5.5. Newborn fennecs show a metabolic response to cold from the first day of life.

     

Myofibril and sarcoplasmic reticulum changes during muscle development: Activity vs inactivity

• 1.1. The purpose of this study was to determine whether biochemical changes of skeletal muscle that occur as a result of exercise in young rats persist into adulthood.
• 2.2. Littermates (10 days old) were assigned to a 3, 6 and 12 week control or training group. In addition, a rest-exercise group (R-E) and exercise-rest (E-R) group were included.
• 3.3. The rest-exercise and exercise-rest rats were maintained for the 12 weeks with the first 6 weeks being either rest or exercise and the condition reversed during the last 6 weeks of the experiment.
• 4.4. Myofibril ATPase activity of rat plantaris increased from the 10d to 12 week animals (P < 0.05). As anticipated, training resulted in a lowered activity at 6 and 12 weeks compared to controls.
• 5.5. The Ca²⁺ uptake and Ca²⁺-ATPase activity of the sarcoplasmic reticulum followed a similar pattern.
• 6.6. With regard to the exercise-rest rats, the myofibril and SR ATPase activities at 12 weeks were comparable to the 12 weeks control rats.
• 7.7. The rest-exercise group approximated the 12 week training group with regard to myofibril and SR ATPase activities (P > 0.05).
• 8.8. The results suggest that the training adaptations that occur during development of skeletal muscle return to normal, when training ceases in the adult rat.
• 9.9. Furthermore, animals that started to train prior to puberty do not have a greater capacity to adapt than animals which initiated training during adulthood.

     

Differential responses in cyclic GMP formation induced by excitatory amino acids (EAA) and sodium nitroprusside (SNP) in various regions of the brain and of rats of varied age

• 1.1. Sodium nitroprusside (SNP, 100 μM) caused a rapid and great increase of formation of cGMP in rat cerebellar slices. This effect was not blocked by l-nmma (a NO synthetase inhibitor) or antagonists of the NMDA receptor complex (e.g. AP₅ or MK 801).
• 2.2. Similarly, NMDA (100 μM) and glutamate (I mM) caused a rapid but less significant increase of cGMP formation. This increase was blocked by NMDA receptor complex blockers (e.g. AP₅, MK801 and kynurenate), and l-NMMA and l-nitroarginine.
• 3.3. In rats aged 12 days, both NMDA and kainate (at 100 μM) caused significantly increased levels of cGMP in the cerebellum, pons and medulla areas, whereas no significant alterations were found in the cerebral cortex, hippocampus or midbrain areas.
• 4.4. NMDA (100 μM) and SNP (300μM) induced greater increases of cGMP in cerebellar slices in young (aged 13 days) animals than older ones of either sex. This effect decreased greatly after 35 days of age. In adult (2 months) animals the effect of NMDA had virtually disappeared whereas SNP was barely significantly present.
• 5.5. Our results suggest that brain region and age, but not sex, affected formation of cGMP induced by excitatory amino acids (EAA) and SNP. Furthermore, endogenous NO production is required by EAA, but not by SNP, in the formation of cGMP.

     

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.

     

Oxygen uptake in larval bollworms (Heliothis zea) infected with the fungus Nomuraea rileyi

• 1.1. Healthy 6- to 12-day-old Heliothis zea (bollworm) larvae showed a mean oxygen uptake of 3.1 μl O₂/mg body wt per hr.
• 2.2. Similar larvae infected with the fungus Nomuraea rileyi had a mean uptake of 4.01 μl O₂/mg per hr.
• 3.3. The weights of the two groups of insects did not differ.
• 4.4. T-test showed a significant (P < 0.01) difference in oxygen uptake between healthy and infected larvae.

     

A comparative study of vertebrate eye lens crystallins using isoelectric focusing and densitometry

• 1.1. The crystallin proteins of numerous species belonging to different classes of vertebrates have been studied.
• 2.2. Species-specific crystallin patterns are revealed which unequivocally characterize the different species.
• 3.3. A marked variability in the number and percentage of alpha-, beta- and gamma-crystallins were found in the various species.
• 4.4. The gamma-crystallin family, with a meagre number of common bands, has proved to be most representative of the species. The beta-crystallins, with their greater number of common bands, have been best preserved throughout vertebrate evolution.
• 5.5. From the similarity coefficient matrix a dendrogram is drawn up, a visual phylogenetic summary of the interrelationships between the vertebrates considered.
• 6.6. In the Discussion, other aspects are considered, such as lens morphology, functionality, animal age, post-synthetic modifications and genetic factors.

     

In vitro substrate utilization for lipid synthesis in liver explants from hyperthyroid chickens

• 1.1. Indian River male broiler chickens growing from 7 to 28 days of age were fed diets containing 12, 18, 24 and 30% protein + 0 or 1 mg triiodothyronine (T₃)/kg of diet to study energetic costs of lipogenesis and the use of various substrates for in vitro lipogenesis.
• 2.2. De novo lipid and CO₂ production were determined in the presence of [1-¹⁴C]pyruvate, [2-¹⁴q]pyruvate, [3-¹⁴C]pyruvate, [2-¹⁴C]acetate and [U-¹⁴C]alanine.
• 3.3. Oxygen consumption was determined in mitochondrial preparations to estimate the energetic costs in expiants synthesizing lipid.
• 4.4. Radiolabeled CO₂ derived from [1-¹⁴C]pyruvate was used as an estimate of coenzyme A availability in liver expiants. Lipids derived from [2-¹⁴C]pyruvate, [2-¹⁴C]acetate and [U-¹⁴C]alanine estimate relative substrate efficiency.
• 5.5. Labeled CO₂ production from [1-¹⁴C]pyruvate was greatest in that group fed a 12% protein diet and least in the group fed a 30% protein diet.
• 6.6. In addition, T₃ increased CO₂ production from [1-¹⁴C]pyruvate.
• 7.7. The production of ¹⁴CO₂ from the second carbon of pyruvate or acetate was increased by T₃.
• 8.8. The low-protein diet (12% protein) increased (P <0.05) lipogenesis.
• 9.9. Adding T₃ to the diets decreased carbon flux into lipid from all substrates, but increased CO₂ production from all substrates without changing stage 3 and 4 respiration rates in mitochondrial preparations.
• 10.10. These observations imply that coenzyme A availability may have regulated de novo lipogenesis in the present study.
• 11.11. It was also concluded that previously noted effects of T₃ on intermediary metabolism may involve metabolic pathways that do not involve changes in mitochondrial function.

     

Regulation of polyamine synthesis in chicken liver

• 1.1. Insulin injected into fasting chickens promotes a marked increase of liver S-adenosylmethionine decarboxylase, with maximum activity 6 hr after administration.
• 2.2. The apparent half-life of the enzyme is 22 min.
• 3.3. Ornithine and putrescine in vivo give rise only to small modifications in enzyme activity; spermidine and spermine induce the enzyme.
• 4.4. Putrescine activates the decarboxylase in vitro. Spermidine and spermine, however, appear to be inhibitors.
• 5.5. Insulin promotes a decrease of arginine, ornithine and all other amino acids, except for proline which increases. Putrescine is enhanced, spermidine slightly decreased and spermine unchanged.
• 6.6. Levels of polyamines are significantly altered by administration of methionine which promotes synthesis of spermidine, and by spermine which blocks transformation of putrescine into spermidine.