Alcohol dehydrogenase ontogeny and liver maturation in the leopard danio,brachydanio nigrofasciatus

• 1.1. The tissue specificity and ontogeny of alcohol dehydrogenase (ADH) are reported for the leopard danio, Brachydanio nigrofasciatus.
• 2.2. Of the seven adult tissues assayed (eye, brain, kidney, liver, ovary, skeletal muscle and stomach), only liver extracts showed ADH activity.
• 3.3. The activation of the Adh locus is correlated with the first functioning of liver. It is suggested that the state of liver cell differentiation may be the stimulus necessary for Adh locus expression.

     

NAD(P)H dehydrogenase from rabbit and rat liver: Purification and some properties

• 1.1. NAD(P)H dehydrogenase from rabbit liver was purified to electrophoretic homogeneity using a procedure also found applicable for the rat liver enzyme.
• 2.2. Rabbit and rat liver enzymes showed different behaviour in isoelectric focusing and different K_m values and turnover numbers.
• 3.3. Both enzymes were inhibited to similar extents by warfarin.
• 4.4. The rabbit enzyme is composed of two subunits of mol. wt 27,000 and contained 1 FAD group per subunit.
• 5.5. Some absorption and circular dichroism properties of the rat enzyme are shown.

     

Soluble cyclic amp-dependent protein kinases from chick liver: Effects of NaCl and heat-stable modulator

• 1.1. Two cyclic AMP-dependent protein kinases—Fraction I and II—have been isolated from chick liver soluble preparation on DEAE-cellulose.
• 2.2. Both fractions have an apparent K_m for ATP of 2 × 10⁻⁶M, are stimulated maximally by 5 × 10⁻⁸ M cyclic AMP and phosphorylate mainly basic proteins—histone and protamine.
• 3.3. They exhibit various pH values for optimal activity and show differences with respect to both sensitivity to NaCl and substrate specificity.
• 4.4. The heat-stable protein modulator inhibits the cyclic AMP-dependent protein kinase activity of both fractions, but with cyclic GMP one kinase is stimulated and the other inhibited.
• 5.5. Slight differences in histone triggered holoenzyme dissociation as well as the lack of difference between their ability for subunit reassociation do not allow to classify these isozymes as protein kinases of Type I and II, according to Corbin et al. (1975).

     

The effect of hypoxia on carbohydrate metabolism in flounder (Platichthys flesus L.)—I. Utilization of glycogen and accumulation of glycolytic end products in various tissues

• 1.1. Resting oxygen consumption at 10°C did not change from normoxia (150 mm Hg) down to an oxygen tension of 55 mm Hg for the flounder, Platichtys flesus.
• 2.2. Flounders exposed to hypoxia showed increased levels of blood glucose and lactate, dependent on the degree of hypoxia.
• 3.3. Due to hypoxia glycogen was depleted in the liver and swimming muscle but in the heart there was no significant change.
• 4.4. Liver glucose increased after 7 hr of hypoxia. Heart and muscle glucose did not change but the absolute glucose concentration in the heart was five times higher than in the muscle.
• 5.5. There is a transient accumulation of lactate in heart, liver and kidney after 7 hr of hypoxia while lactate accumulation in the swimming muscle is significant only after 21 hr of hypoxia.
• 6.6. Succinate only accumulated in the liver while alanine accumulated in muscle, heart and liver.

     

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.

     

Effects of temperature and food on the free amino acids in tissues of roach (Rutilus rutilus L.) and rudd (Scardinius erythrophthalmus L.)

• 1.1. In liver and muscle the concentrations of free amino acids (FAA) are highest in fish maintained at low temperature and fed mealworms. These effects are more pronounced in roach than in rudd.
• 2.2. In the liver alanine, glycine and glutamate are the dominant FAA but proline increases in mealworm-fed animals.
• 3.3. In muscle, histidine and glycine dominate, except that a mealworm diet leads to an increase in the concentration of proline and to a concomitant decrease in the concentration of glycine.
• 4.4. Starvation leads to a reduction of total FAA content but to relative increases of lysine and histidine. These two FAA can serve as indicators of the general state of nutrition of roach and rudd.
• 5.5. The molar ratio [gly]/[his] is strongly correlated with temperature, decreasing with an increase in the temperature to which the animals had been exposed prior to capture.
• 6.6. The patterns of free and bound amino acids diverge more widely in these species than in mammals which reflects the greater dependence of the FAA pools of fish on intrinsic and extrinsic factors.
• 7.7. The concentrations of histidine in the FAA pools of muscle and in food proteins are strongly correlated.

     

AMP-deaminase from human skeletal muscle. Subunit structure,amino-acid composition and metal content of the homogenous enzyme

• 1.1. Homogenous human skeletal muscle AMP-deaminase was obtained by chromatography on phosphocellulose.
• 2.2. Native enzyme molecular weight was 290,000, while a value of 71,000 was found for the subunit molecular weight.
• 3.3. No distinct differences were found in amino-acid composition of human skeletal muscle AMP-deaminase as compared with other vertebrate enzymes.
• 4.4. Human muscle AMP-deaminase contains about 2g-atom of zinc per 280,000; considerable amounts of calcium and magnesium were also found.

     

Malate dehydrogenase isoforms from ribbed mussel (Modiolus demissus) gill tissue

• 1.1. The malate dehydrogenase (MHD) activity from the ribbed mussel gill is polymorphic with two distinct mitochondrial forms (M1 and M2) and five forms that could be resolved from cytosolic extracts (C1 to C5) by DEAE-cellulose chromatography and starch gel electrophoresis.
• 2.2. Two of the cytosolic forms (C3 and C4) may represent interchangeable conformational states.
• 3.3. With kinetic analysis there appear to be three distinct cytosolic forms (C1, C2 and C3–C4), with C2 possibly behaving as a heterodimer.
• 4.4. The identity of C5 is uncertain.
• 5.5. The forms isolated from the mitochondria (M1 and M2) exhibited lower apparent K_ms for oxaloacetate (OAA) than the cytosolic forms.
• 6.6. For all isozymic forms, the apparent K_ms for OAA increased as the pH increased between pH 6 and 9
• 7.7. Increasing the salt concentration raised the K_m for OAA for all forms.
• 8.8. The mMDHs were more sensitive to inhibition by NaCl than the cMDHs.
• 9.9. Representative cMDH (C1) and mMDH (M2) isozymes exhibited substrate inhibition by high concentrations of OAA with the mMDH possessing lower K_is for substrate inhibition than the cMDH at each pH tested.
• 10.10. Differences and similarities in K_m app. for OAA at the different pHs and salt concentrations indicated that C1, C2 and C3–C4 and C5 were distinct forms, that M1 and M2 were distinct but very similar to each other, and that C1, C2, C3–C4 and C5 were distinct from M1 and M2.

     

Phylogeny and ontogeny of the phosphoglycerate mutases—I. Electrophoretic phenotypes of the glycerate-2,3-P2 dependent phosphoglycerate mutase in vertebrates

• 1.1. The electrophoretic phenotype of phosphoglycerate mutase in tissues from different classes of vertebrates at several stages of development have been analyzed on cellulose acetate.
• 2.2. Mammals, reptiles, amphibians and fish show a common three-banded isozyme pattern.
• 3.3. The three isozymes vary in their relative distribution from tissue to tissue and during growth.
• 4.4. In birds electrophoretically distinguishable phosphoglycerate mutase isozymes have not been detected.
• 5.5. The results support a genetic basis for the phosphoglycerate mutase isozymes and suggest that gene duplication may have occurred early in vertebrate evolution.

     

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.

     

Age-related changes in total dna and rna and incorporation of uridine and thymidine in rat liver,kidney and spleen

• 1.1. Total content of DNA and RNA in liver, kidney and spleen were measured in young and aged rats. At the same time the incorporation of [¹⁴C]thymidine, a DNA precursor, and [³H]uridine, an RNA precursor, were also determined.
• 2.2. Changes in total organ DNA and RNA correlated with sexual maturation as did incorporation of precursors.
• 3.3. Young animals have more DNA per organ relative to RNA. with kidney and spleen DNA showing a decrease between maturity and senescence.
• 4.4. However, liver RNA increases with age. a change probably due to decreased catabolism of RNA since [³H]uridine uptake decreases.
• 5.5. Liver polyploid differentiation, and [¹⁴C]thymidine and [³H]uridine uptake, are correlated.
• 6.6. In kidney, incorporation of [³H]uridine is inversely related to [¹⁴C]thymidine incorporation.

     

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⁺.

     

IDentification of a microsomal retinoic acid synthase as a microsomal cytochrome P-450-linked monooxygenase system

• 1.1. To characterize an enzyme which metabolizes retinal in liver microsomes, several properties of the enzymatic reaction from retinal to retinoic acid were investigated using rabbit liver microsomes.
• 2.2. The maximum pH of the reaction in the liver microsomes was 7.6.
• 3.3. The K_m and V_max values for all-trans, 9-cis and 13-cis-retinals were determined.
• 4.4. The reaction proceeded in the presence of NADPH and molecular oxygen.
• 5.5. The incorporation of one atom of molecular oxygen into retinal was confirmed by using oxygen-18, showing that the reaction comprised monooxygenation, not dehydrogenation.
• 6.6. The monooxygenase activity was inhibited by carbon monoxide, phenylisocyanide and antiNADPH-cytochrome P-450 reductase IgG, but not by anti-cytochrome b₅ IgG.
• 7.7. The enzymatic activity inhibited by carbon monoxide was photoreversibly restored by light of a wavelength of around 450 nm.
• 8.8. The retinal-induced spectra of liver microsomes with three isomeric retinals were type I spectra.
• 9.9. The microsomal monooxygenase activity induced by phenobarbital or ethanol were more effective than that by 3-methylcholanthrene, clotrimazole or β-naphthoflavone.
• 10.10. These results showed that the monooxygenase reaction from retinal to retinoic acid in liver microsomes is catalyzed by a cytochrome P-450-linked monooxygenase system.

     

Effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail

• 1.1. The effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail were investigated.
• 2.2. The level of liver NAD was not affected by niacin deficiency whereas the level of pectoral muscle NAD was markedly reduced.
• 3.3. In all dietary treatments the liver had the highest turnover rates of proteins, heart and brain had intermediate rates, and pectoral muscle had the lowest rates.
• 4.4. Relative turnover rates of proteins in all tissues (particularly pectoral muscle) of the niacin deficient group were significantly higher than those of pair-fed control group, although there were no significant differences in turnover rate between pair-fed control and control groups.
• 5.5. The high turnover rate of proteins in niacin deficiency was primarily attributed to enhanced degradation rate of proteins rather than enhanced synthesis rate of proteins.
• 6.6. Optical density scanning (or densitometric) of water-soluble pectoral muscle proteins separated by isoelectric focusing revealed several additional minor protein bands between major protein bands in the niacin deficient group which were more pronounced in the acidic region of the gel.
• 7.7. These results suggest that proteins with a low pI value in pectoral muscle of the niacin deficient animal are highly sensitive to protein degradation.

     

Temperature acclimation in amphibians: Changes in lactate dehydrogenase activities and isoenzyme patterns in several tissues from adult Discoglossus pictus pictus (Otth.)

• 1.1. The effect of cold (8 ± 2°C) acclimation on the lactate dehydrogenase activities and isoenzyme patterns from sartorius muscle, liver, heart and brain of adult Discoglossus pictus pictus (Otth.) was studied.
• 2.2. Two groups of animals were studied: one set of animals was trapped in October and another set in December. In both cases some of the animals were sacrificed upon collection and some others subjected to 5 months of acclimation at 8 ± 2°C before being sacrificed for analysis.
• 3.3. A general trend towards a decrease in LDH specific activity was observed during cold acclimation. The magnitude of change, but not the direction, depends on both the tissue examined and the season at which the experiment was initiated.
• 4.4. A complex LDH isoenzyme reorganization was also found in liver, heart and brain. In liver from Experiment 1 and in heart from both experiments, a relative maintenance in M-type LDH activity during cold acclimation was observed. However, in brain there was a relative maintenance of LDH₃ activity in both experiments.
• 5.5. The low behavioral activity (and its metabolic consequences) and the existence of an intrinsic annual rhythm in D. pictus metabolism are suggested as responsible for the observed enzymatic changes.

     

The effect of hypoxia on carbohydrate metabolism in flounder (Platichthys flesus L.)—II. High energy phosphate compounds and the role of glycolytic and gluconeogenetic enzymes

• 1.1. ATP, ADP, AMP, energy charge potential and total adenylates in heart, kidney and muscle are relatively unaffected by environmental hypoxia. In the liver, hypoxia causes a 90% drop in ATP, a rise in ADP and AMP, and a drop in energy charge potential and total adenylates. In the muscle tissue ATP concentration is stabilized by a large creatine phosphate pool.
• 2.2. Hexokinase activity in the heart is 20 times higher than in the swimming muscle, and thus the heart has a high potential for utilizing exogenous glucose as an anaerobic substrate.
• 3.3. The role of creatine phosphate in regulating muscle glycolysis is discussed on background of the strong inhibition of muscle phosphofructokinase by physiological concentrations of creatine phosphate.
• 4.4. Flounder heart has a dominating M-type lactate dehydrogenase which is identical to the muscle enzyme by electrophoretic and kinetic criteria. This improves the anaerobic capabilities of the flounder heart compared to other fish hearts.
• 5.5. Both liver and kidney have high activities of the gluconeogenetic enzymes glucose-6-phosphatase, fructose-1,6-diphosphatase, and phosphoenolpyruvate carboxykinase and both are capable of synthesizing glucose from [¹⁴C]lactate. Because of more favorable energy conditions in the kidney this organ may substitute the liver as a gluconeogenetic organ during hypoxia.

     

Variations in tissue reserves,plasma metabolites and pancreatic hormones during fasting in immature carp (Cyprinus carpio)

• 1.1. Immature carp were subjected to 2-month fasting periods. Mobilization of reserves in liver and muscle, and the energy contribution of each reserve were studied. Changes in plasma glucose, amino acids, insulin and glucagon levels were determined throughout the experiment.
• 2.2. No changes were observed in plasma glucose, insulin or glucagon at 19 days of fasting, but plasma amino acids increased. At 50 days of fasting, both plasma glucagon and amino acids increased, liver glycogen decreased and muscle proteolysis began.
• 3.3. Between 50 and 67 days of fasting, plasma glucose and insulin decreased significantly, while glucagon and amino acids continued to increase. Strong muscular proteolysis was observed while liver glycogen stabilized.
• 4.4. The contribution of each reserve in liver and muscle to energy production throughout fasting is considered.

     

Creatine kinase from the electric organ of Electrophorus electricus (L.)—isozyme analysis

• 1.1. Creatine kinase (CPK) isozymes of extracts from the electric organ, dorsal muscle and brain of Electrophorus electricus (L.) were analysed with Cellogel electrophoresis. A single component corresponding to the MB-form was obtained for both electric organ and the dorsal muscle. The BB-form was present in the brain extract.
• 2.2. Upon acetone fractionation of the aqueous extract of electric organ, the final fraction was submitted to gel filtration and presented a single peak of CPK activity.
• 3.3. Characterization of this fraction by thin-layer gel filtration indicated an apparent molecular weight of 80,000 which corresponds to the enzyme dimeric structure.
• 4.4. The implications of this finding with the muscular origin of the electric organ are discussed.

     

Lysosomal acid deoxyribonuclease from vervet monkey livers—II: Enzymic properties

• 1.1. Primate liver lysosomal acid DNase is an endonucleolytic enzyme.
• 2.2. The enzyme has both 3'- and 5'-nucleotidohydrolase activities.
• 3.3. The oligonucleotides produced by DNase are polymers mainly about 30 mononucleotides long.
• 4.4. The Arrhenius plot shows a discontinuity with a transition temperature at 47°C, with an activation energy of 107 kJ/mol below and 67 kJ/mol above this temperature.
• 5.5. The activation enthalpy is 104kJ/mol and the entropy −0.498 kJ/mol/K.
• 6.6. The enzyme is subject to substrate inhibition and the K_m value is 159 × 10⁻³mM DNA-P.

     

Phylogeny and ontogeny of the phosphoglycerate mutases—II. Characterization of phosphoglycerate mutase isozymes from vertebrates by their thermal lability and sensitivity to the sulfhydryl group reagents

• 1.1. To characterize the three phosphoglycerate mutase (PGM) isozymes present in vertebrates (types M, B and MB) their sensitivity to the reagents of the sulfhydryl groups and to heat treatment has been studied.
• 2.2. In mammals and reptiles type M PGM is not affected by the —SH group reagents, type MB PGM is inhibited about 50% and type B PGM is fully inhibited. Types B and MB PGM show greater heat lability than type M PGM.
• 3.3. In amphibians and fishes PGM isozymes do not differ in their sensitivity to the —SH reagents.
• 4.4. The results strongly support the homodimeric and heterodimeric structure suggested for PGM isozymes and favour their genetic origin.