Human GMP synthetase

• 1.1. The specific activity of GMP synthetase was measured in several human tissues and found to be highest in cultured skin fibroblasts, followed by bone marrow, leukocytes, erythrocytes. placenta, and liver.
• 2.2. The enzyme from fibroblasts was purified approximately 50-fold by ammonium sulfate fractionation and gel filtration.
• 3.3. The K_m values were determined to be 4.9μM for XMP, 270μM for ATP. and 340 μM for glutamine.
• 4.4. Ammonium sulfate could replace glutamine as the amino donor but was much less efficient.
• 5.5. The enzyme was specific for ATP as the energy source.
• 6.6. Unlike the calf thymus enzyme, the human enzyme has no requirement for a reduced sulfhydryl compound.
• 7.7. Human GMP synthetase is inhibited by ATP, dATP, azaserine, and hydroxylamine.

     

Regulation of glutamine catabolism in the perfused guinea-pig liver in relation to ureogenesis and gluconeogenesis

• 1.1. L-Glutamine conversion into ammonia, urea and glucose by the perfused liver of 48 hr starved guinea-pigs was concentration dependent attaining the maximal rate at 4 mM.
• 2.2. The activity of glutaminase I (EC 3.5.12), measured in isolated liver mitochondria was high enough to account for the observed rate of ammonia, urea and glucose formation by the perfused liver. Neither NH4C1 (5 mM) nor aminooxyacetate (0.5 mM) affected the rate of glutamine conversion into glutamate by isolated liver mitochondria.
• 3.3. Gluconeogenesis and ureogenesis from glutamine was inhibited by octanoate, Dt-3-hydroxybutyrate, aminooxyacetate, ethanol and p-hydroxyphenylpyruvate while ammonia formation was stimulated by aminooxyacetate. 2,4-Dinitrophenol stimulated the rate of the formation of all three metabolites from glutamine.
• 4.4. The major changes induced by aminooxyacetate, as determined in livers perfused with glutamine and stopped by freeze-clamping technique, consisted in a decrease in the content of ATP, aspartate and malate and in a slight increase in the content of glutamate.
• 5.5. Glutamine is an effective precursor of phosphoenolpyruvate in isolated liver mitochondria. Its formation was inhibited by octanoate and by DL-3-hydroxybutyrate.
• 6.6. The data are discussed in terms of regulation of glutamine catabolism in liver with emphasis on ureogenesis and gluconeogenesis.

     

Regulation of spermidine transport in l1210 cells

• 1.1. 1 mM 2-amino isobutyric add (AIB), glutamine or asparagine when preincubated for 3 hr with L1210 cells promoted a marked increase in the rate of spermidine uptake.
• 2.2. Cycloheximide also increased the transport rate and completely prevented the increase due to AIB.
• 3.3. Trifluoperazine and iso-H7 inhibited the uptake of spermidine, much less the uptake of AIB.
• 4.4. Adenosine promoted an increase in the uptake of AIB, a decrease in that of spermidine.
• 5.5. Hypotonic stress also increased the rate of spermidine transport. This modification was only partially prevented by cycloheximide.
• 6.6. Okadaic arid had no effect on this increase, whereas it prevented the increase of ODC activity.

     

Distribution of tetracycline in haemolymph and tissue of all stages of corn earworm (Heliothis zea boddie)

• 1.1. Insects reared on tetracycline containing diets had lower mortality, but required more time to reach the pupal stage.
• 2.2. Tetracycline diffuses and dissipates readily into and out of insect blood and tissues.
• 3.3. It reduces biosynthesis of high molecular weight proteins in blood.
• 4.4. Apportionment of tetracycline in the pupal tissue is uneven.

     

Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis (L.)

• 1.1. Anaerobic energy metabolism was investigated in different organs of Mytilus edulis and the whole animal.
• 2.2. Succinate accumulates to high levels in most organs but remains low in the hemolymph.
• 3.3. After 16 hours propionate accumulation is observed in all organs. Experimental evidence is not sufficient yet to point out organs that produce more propionate than others.
• 4.4. Acetate is a minor end product.
• 5.5. Acetate and propionate are found in the hemolymph in amounts equal to those in the organs.
• 6.6. Animals incubated in oxygen-free seawater accumulate more end products than animals exposed to air, in the form of volatile fatty acids that are excreted into the incubation water.
• 7.7. Alanine and glutamine increase in the posterior adductor muscle. Aspartate decreases in the total animal, posterior adductor muscle and gills, while in the hemolymph decrease in alanine, asparagine, serine, threonine and proline are observed.

     

Effect of pH on metabolism of the glutamine carbon skeleton by renal cortical mitochondria

• 1.1. To determine the effect of altered acid-base homeostasis on the intramitochondrial metabolism of the glutamine carbon skeleton ¹⁴CO₂ production from [U-¹⁴C]glutamine by isolated rat renal cortical mitochondria was measured.
• 2.2. Mitochondria from rats with chronic metabolic acidosis either showed no change or diminished ¹⁴CO₂ production in comparison with pair fed controls.
• 3.3. By contrast, when the pH of the medium incubating mitochondria from normal rats was manipulated (pH 7.0, 7.4, 7.7), ¹⁴CO₂ production was clearly altered, but the direction and magnitude of the change depended on the glutamine concentration used (0.5 or 10.0 mM).
• 4.4. Mitochondria produced significant quantities of ¹⁴CO₂ when [1,4 ¹⁴C]succinate was used as substrate, indicating that ¹⁴CO₂ production from glutamine does not originate solely from the decarboxylation of α KG.
• 5.5. Thus chronic acidosis and pH, per se, affect intramitochondrial glutamine carbon skeleton metabolism in different fashions, but the specific mechanism cannot be elucidated using ¹⁴CO₂ production from [U-¹⁴C]glutamine.
• 6.6. Additional studies directly quantitating the metabolic products of glutamine have confirmed these findings and more precisely defined the sites of metabolic alteration.

     

Larval Musca domestica lipophorin biosynthesis

• 1.1. Larval Musca domestica lipophorin biosynthesis was studied in vitro.
• 2.2. The newly synthesized lipophorin has a density a little lower than the circulating lipophorin after 1 hr of incubation. After 3 hr of incubation the fat body cells transfer lipids to the lipophorin that attains the density of circulating lipophorin.
• 3.3. The lipophorin synthesized in vitro is identical to circulating lipophorin in density and in electrophoretical behavior.
• 4.4. However these two molecules must have differences since the circulating lipophorin transfers lipids to fat body cells while the synthesized in vitro does not.
• 5.5. The biosynthesis of Musca lipophorin shows differences with the Manduca sexta lipophorin biosynthesis.

     

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Analysis of peptide biosynthesis in the neurointermediate lobe of Xenopus laevis using high-performance liquid chromatography: occurrence of small bioactive products

• 1.1. Ppeitde biosynthesis in neurointermediate lobes of black adapted Xenopus laevis was studied using pulse-chase incubation and reversed-phase, high-performance liquid chromatographic analysis.
• 2.2. During the pulse period one major product was synthesized, which was subsequently converted to 12 chase peptides, suggesting a precursor-product mode of biosynthesis for this tissue.
• 3.3. Chase peptides I, II and IV possessed high melanotropic activity. Alpha-MSH did not appear to be among the chase peptides. Peptide II had also high corticotropic activity.
• 4.4. Peptides I and II are probably small, since they were TCA-soluble and ran faster on acid-urea gels than α-MSH. They may, however, well be structurally related to this latter hormone.

     

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Drosophila mettleri and senita cactus alkaloids: Fitness measurements and their ecological significance

• 1.1. Drosophila mettleri have been found feeding but not breeding on decaying stems of senita cactus, the normal host for Drosophila pachea.
• 2.2. Alkaloids were extracted from senita stems and used in tests of egg-to-adult viability, developmental rate, and adult longevity.
• 3.3. The results show that developmental rate is not appreciably affected by senita alkaloids.
• 4.4. In general, D. mettleri was less affected by the alkaloids with respect to egg-to-adult viability and adult longevity than D. pachea at concentrations which are fatal to other desert Drosophila.
• 5.5. Tolerance to alkaloids gives D. mettleri an ecological advantage.

     

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Adenosine and deoxyadenosine metabolism in lymphocytes from horses,Equis cabalus

• 1.1. The toxic effects of adenosine and deoxyadenosine on lymphocytes from horses were evaluated.
• 2.2. Mitogen-stimulated peripheral blood lymphocytes (PBL) were found to be more sensitive to the inhibitory effects of adenosine than were lymphocytes from spleen, lymph node and thymus.
• 3.3. Adenosine deaminase activity was approximately 10 times lower in horse lymphoid tissue in comparison to that found in human lymphoid tissue. In horse, the highest activity was in spleen while the lowest activity was in thymus.
• 4.4. Adenosine inhibition of mitogenesis in PBL was prevented by uridine or cytidine, suggesting pyrimidine starvation as the major mechanism for adenosine toxicity.
• 5.5. Deoxyadenosine inhibition of mitogenesis in PBL was not prevented by the addition of various ribo- or deoxyribonucleosides. This and the finding that treated cells show no increase in deoxyATP suggest that some other mechanism for deoxyadenosine toxicity other than deoxyATP inhibition of ribonucleotide reductase operates in horse PBL.

     

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Kinetic analysis of reversible closed bicyclic enzyme cascades covering the whole course of the reaction

A kinetic analysis of the closed bicyclic enzyme cascades is presented.

• 1.1. It includes the dependence on time from the onset of the reaction, of the concentration of the modified and unmodified enzyme species involved and the time course equations of the modificational fractions of the interconvertible enzymes.
• 2.2. The transient phase equations obtained allow the definition of new regulatory modification properties.
• 3.3. The expressions for concentrations of the unmodified and modified forms of the interconvertible enzymes, as well as those of the fractional modifications in the steady state are derived as particular cases of the general equations.
• 4.4. These steady state expressions coincide with those obtained by other authors.
• 5.5. The analytical results obtained are discussed in relation to the Escherichia coli glutamine syntethase cascade.

     

Aspects of iron metabolism in Bifidobacterium bifidum var. pennsylvanicus

• 1.1. Btfidobacterium bifidum var. Pennsylvanias requires ferrous iron for growth, and cannot utilize ferric iron even in the presence of siderophores.
• 2.2. Acid production by the microorganisms is dependent in part on iron content of the medium.
• 3.3. Heme and heme-containing proteins inhibit the microbial growth, and it is proposed that this is in part responsible for the change in the infant's intestinal flora upon weaning.
• 4.4. Bacterial growth inhibition brought about by heme cannot be restored by heme biosynthesis intermediates, and known heme biosynthesis inhibitors have no effect on bacterial growth. The basis for heme-induced microbial growth inhibition remains unclear.

     

Effect of aminooxyacetate and α-ketoglutarate on glutamate deamination by rat kidney mitochondria

• 1.1. Glutamate dehydrogenase flux by rat kidney mitochondria incubated with 1 mM glutamine plus 2–3 mM glutamate was stimulated by aminooxyacetate. This effect was inhibited by α-ketoglutarate.
• 2.2. Studies with intact mitochondria and mitochondrial sonicates revealed a linear inverse relationship between glutamate deamination and α-ketoglutarate levels.
• 3.3. The data revealed that α-ketoglutarate is a competitive inhibitor of glutamate dehydrogenase with an apparent K_i of 0.6mM.
• 4.4. The data suggest that aminooxyacetate stimulates glutamate deamination by a mechanism mediated by α-ketoglutarate.

     

Effect of lindane on phosphatidylinositol synthesis by cerebral cortex after acute and subchronic treatment

• 1.1. The incorporation of myo-[2-³H]inositol into phosphatidylinositols was unmodified in brain cortex miniprisms from convulsant rats.
• 2.2. However, the incorporation had increased by 300–400% in non convulsant rats which had received the same amount of lindane at a lower concentration.
• 3.3. This result suggests that phosphatidylinositols are implicated in the convulsion syndrome.
• 4.4. Experiments with lindane added in vitro were performed with both subchronically lindane intoxicated and untreated rats.
• 5.5. The results show an interesting lack of parallelism.
• 6.6. This might indicate the development of some resistance to the effects of lindane, possibly as the result of complex compensatory changes in inositol lipid biosynthesis.

     

Osmotically-induced volume changes in isolated cells of a pond snail

• 1.1. To assess whether the stretch-activated (SA) channels in snail cells could contribute to osmoregulation, information is needed about the behaviour of the cells under anisosmotic conditions.
• 2.2. Cells of Lymnaea stagnalis were therefore examined during acute hyposmotic stress (HOS).
• 3.3. Kidney, heart and neuronal cells (monitored photographically) swelled less than expected for strictly semipermeable cells, but exhibited no regulatory volume decrease.
• 4.4. Long-term viability of the cells was not compromised following acute hyposmotic stress.
• 5.5. Quinidine, which blocks SA channels in Lymnaea, intensified stress-induced swelling most markedly in kidney cells.
• 6.6. The data can, however, be explained without invoking recruitment of SA channels.

     

Effect of diet and essential amino acids gavage on young rat amino acid metabolism enzymes

• 1.1. The effects of a high-fat, high-energy diet and essential plus semi-essential amino acid gavage on pup rats have been studied (60–65 animals).
• 2.2. The activities of alanine transaminase, adenylate deaminase, glutamine synthetase and serine dehydratase have been tested in liver and muscle.
• 3.3. Plasma was used for the estimation of proteins, urea, amino acids, glucose, lactate, 3-hydroxy-butyrate and acetoacetate.
• 4.4. Liver and muscle glutamine synthetase activities are increased by diet and gavage administered. Hepatic serine dehydratase is inhibited by a cafeteria diet but activated by amino acid gavage. Adenylate deaminase is inhibited by diet and gavage in the liver, but gavage does not affect this enzyme activity in muscle. Liver alanine transaminase is increased by the diet; in the muscle, cafeteria diet and amino acid gavage showed the highest values for this enzyme.
• 5.5. In the plasma, the increase in lactate produced by the diet is inhibited by the amino acids provided. Cafeteria-fed pups showed lower urea levels and higher 3-hydroxybutyrate concentrations in the plasma.
• 6.6. Intracellular glucose is diminished by cafeteria diet. In contrast, the blood cell amino acid concentration increases with diet and gavage supplied.