The role of Zn2+ in pyridoxal phosphate mediated regulation of glutamic acid decarboxylase in brain

• 1.1. γ-Aminobutyric acid, a major inhibitory neurotransmitter in the CNS, is synthesized by glutamic acid decarboxylase which demonstrates an absolute requirement for pyridoxal phosphate.
• 2.2. At physiological concentrations, zinc stimulates the activity of pyridoxal kinase, enhancing the formation of pyridoxal phosphate, which in turn stimulates the activity of glutamic acid decarboxylase.
• 3.3. At pharmacological concentrations, zinc inhibits the activity of glutamic acid decarboxylase without inhibiting pyridoxal kinase.
• 4.4. These results suggest that zinc may play a role in pyridoxal phosphate-mediated regulation of glutamic acid decarboxylase.

     

Some studies on the oxygen affinity of haemoglobin from the dugong

• 1.1. The P₅₀ of dugong haemoglobin at pH 7.4 and 30°C in O.1 M sodium phosphate buffer and 0.1 g % concentration was found to be 6.2 mm Hg.
• 2.2. The Bohr shift in the pH range 6.5–7.0 was −0.48. The value of the Hill coefficient was maximally 1.85.
• 3.3. The heat of combination, for the oxygen-haemoglobin binding reaction was found to be 77.2kjoules/°K per mole.
• 4.4. The results are interpreted as suggesting significant subunit dissociation of dugong haemoglobin at low concentrations.

     

Accumulation,distribution and toxicity of selenium in the adult house fly,Musca domestica

• 1.1. Accumulation and distribution of dietary Se in relation to mortality was investigated in adult house flies.
• 2.2. The midgut preferentially accumulated Se and thereby limited toxicity.
• 3.3. Midgut Se concentrations were from 6- to 107-fold higher than in carcass, and from 15 to 71% of the total Se was associated with midgut.
• 4.4. When dietary levels of Se were raised the midgut saturated at 15 μg Se/g tissue, followed by a rise in carcass levels to greater than 0.5 μg Se/g tissue and increased mortality.
• 5.5. Se levels in lysosomal fractions were from 3- to 50-fold higher than in other subcellular fractions, suggesting that Se is sequestered in lysosomes.
• 6.6. Se added to drinking water was toxic at 4–8 ppm.

     

Seasonal changes of lipids and fatty acids in oyster tissues (Crassostrea virginica) and estuarine particulate matter

• 1.1. Lipid concentration in adductor muscle ranged from 2–68, in visceral mass from 5–28, in mantle and gill from 5–20 and in heart from 27.8–79 mg/g wet tissue. Particulate matter lipids varied from 1.0–2.6 mg/1 of estuarine water.
• 2.2. Neutral and polar lipids ranged from 25–38% of the total lipids in the oyster tissue and from 62–75% of the estuarine particulate organic matter.
• 3.3. Seasonal maxima of lipid concentrations varied among oyster tissues. Peak particulate lipids occurred in November.
• 4.4. It is proposed that seasonal variation in oyster lipids was more related to reproductive cycles than to food lipid supply.

     

A novel inhibitor of glycogen phosphorylase from crayfish hepatopancreas

• 1.1. A novel glycogen phosphorylase inhibitor was partially purified from crayfish hepatopancreas.
• 2.2. The inhibitor was found only in two species of crayfish examined, and not in lobster, fresh and salt water clams, mussels or cockroaches.
• 3.3. The inhibitor is a small protein (M_r = 23,000) which did not show proteolytic activity.
• 4.4. Preliminary kinetic analysis of the inhibitory mechanism indicated that it bound to both glycogen and the glycogen phosphorylase protein.
• 5.5. Inhibitor binding to glycogen resulted in a competitive inhibition pattern with respect to glycogen phosphorylase (inhibition constant of ca 10 μg/ml).
• 6.6. The inhibitor also bound glycogen phosphorylase directly with a binding coefficient of 100 μg/ml resulting in a partially non-competitive inhibition pattern with respect to phosphate.

     

Phosphorylation of a 25,000-dalton protein in slow and fast muscles of the chicken

• 1.1. Protein phosphorylation in intact chicken latissimus dorsi muscle, slow anterior (ALD) and fast posterior (PLD), was compared.
• 2.2. A major difference in [³²P]phosphate incorporation was found between the ALD and PLD in a 25,000-dalton heat soluble protein.
• 3.3. The 25,000-dalton protein was purified from both the ALD and PLD.
• 4.4. The two proteins had similar amino acid composition and both contained approximately 1 mole phosphate per mole of protein.
• 5.5. The difference in their content of radioactive phosphate was determined to be due to faster turnover in the ALD.

     

Laboratory studies on the oxygen consumption of the marine teleost,Lichia amia (Linnaeus, 1758)

• 1.1. The oxygen consumption of the marine teleost, Lichia amia was investigated under controlled laboratory conditions.
• 2.2. The routine oxygen consumption showed a strong circadian rhythm with the fish being mainly active during the light period.
• 3.3. The specific mass exponent (dimension: μg O₂/g/hr) is temperature independent and ranges from 0.27–0.29.
• 4.4. Starving the fish results in a mean decrease in active, routine and standard oxygen consumption of 21%, 24% and 20%, respectively.
• 5.5. Feecling led to an increase in the oxygen consumption of the teleosts, with the mean metabolic rate over the 24 hr that followed, being 58% and 50% higher for fish that had been starved for 162hr and 40 hr, respectively.
• 6.6. Apparent SDA showed some variation and ranged from 6.0 to 35.5%.
• 7.7. The results obtained are generally in agreement with those recorded for other teleosts.

     

Interaction of pyridoxal phosphate with thymidylate synthase: Spectral and equilibrium dialysis studies

• 1.1. Changes in the spectrum of pyridoxal phosphate (PLP) were produced by adding an equimolar amount of native thymidylate synthase, but not by adding denatured enzyme or enzyme modified by sulfhydryl-blocking reagents.
• 2.2. The dissociation constant of the thymidylate synthase-PLP complex determined by equilibrium dialysis was 9 ± 1.6 μM, the maximum number of PLP molecules bound per molecule of native thymidylate synthase was 2.5 ± 0.4, and the Hill coefficient was 0.97.
• 3.3. No evidence of PLP binding was found with denatured thymidylate synthase, and only slight binding was observed when enzyme SH groups were blocked or when the active site was blocked with 5-fluorodeoxyuridylate (FdUMP) and methylenetetrahydrofoliate.
• 4.4. The presence of dUMP, dTMP, or FdUMP interfered with the binding of PLP to thymidylate synthase, and the presence of equimolar amounts of PLP interfered with the binding of dUMP.

     

Adaptation of rainbow trout (Salmo gairdnerii) to sea water: Changes in calcitonin levels

• 1.1. The calcitonin content of the ultimobranchial body (UBB) and plasma levels of calcitonin, calcium and phosphate were measured in rainbow trout (Salmo gairdnerii) following their transfer from fresh to sea water.
• 2.2. The plasma calcium level remained unchanged throughout the experiment while the UBB calcitonin content, plasma calcitonin and plasma phosphate rose significantly during the hours immediately following transfer.
• 3.3. The levels of all three subsequently fall so that, 8–15 days later, a new equilibrium was established with lower than control (fresh water) levels of UBB calcitonin, plasma calcitonin and plasma phosphate.
• 4.4. It would appear, from these data, that calcitonin plays some part in the endocrine regulation of sea water transfer.

     

Essential tyrosyl residues of human placental alkaline phosphatase

• 1.1. Human placental alkaline phosphatase was inactivated with tetranitromethane in a biphasic process.
• 2.2. Spectral and amino acid analysis demonstrated that the inactivation was due to the conversion of tyrosine residues to 3-nitrotyrosine.
• 3.3. The inactivation process showed saturation kinetics.
• 4.4. Protection of the enzyme against tetranitromethane inactivation was afforded by inorganic phosphate.
• 5.5. The binding affinity between the modified enzyme and inorganic phosphate was decreased.
• 6.6. Our results suggest the involvement of tyrosyl residues in the locus of phosphoryl site of the phosphorylated enzyme forms.

     

The high intracellular ph buffering of hagfish (Eptatretus cirrhatus) dental plate retractor muscle cannot be attributed to the known histidine-related compounds present in other vertebrates

• 1.1. Intracellular pH buffering capacity of hagfish (Eplatretus cirrhatus) dental plate retractor muscles is among the highest reported for any vertebrate muscle.
• 2.2. Over 80% of the pH buffering capacity of hagfish retractor and myotome muscle is due to components other than proteins and phosphate.
• 3.3. The muscles have less than 0.5 μmol/g wet weight of l-histidine, and lack l-l-methyl histidine, l-3-methyl histidine and the histidine-containing dipeptides anserine, carnosine and ophidine.
• 4.4. Instead, they contain an unidentified low molecular weight acid-soluble compound to which the high pH buffering capacity can be attributed.

     

Quantitative determination of nucleosides and their phosphate esters—1. The acidic nucleosides, 3-deazauridine and 6-azauridine

• 1.1. A relatively rapid, high-resolution Chromatographic procedure, using mini-columns of DEAE cellulose equilibrated with 10mM sodium phosphate, pH 6.0, is described in sufficient detail to permit ready replication.
• 2.2. This initial paper demonstrates the quantitative separation, using suction, of the acidic nucleosides, 3-deazauridine and 6-azauridine, from their phosphorylated derivatives.
• 3.3. The chemically stable, tritium-labeled nucleosides are eluted from the mini-columns (capacity ≈ 1.8 ml) with 10mM sodium phosphate, pH 6.0; subsequently, the nucleotides are eluted completely with 0.5 M HCl/0.5 M NaCl.
• 4.4. Quantitation is based on liquid scintillation counting of aliquots of the eluates.

     

Phosphate metabolism during muscular contraction in starved frogs (Rana catesbeiana)

• 1.1. The muscle tension and the state of high-energy phosphate metabolism during contraction of the sartorius muscle in frogs (Rana catesbeiana) starved for 1–5 months was studied by in vivo³¹P-NMR spectrometry.
• 2.2. Muscle tension began to decrease after 2-month starvation compared with the control group and decreased to about one-third of the control value after a 5-month starvation.
• 3.3. Muscle contraction induced by electrical stimulation or the use of anaerobic perfusion fluid did not decrease the concentration of creatine phosphate (PCr) or β-ATP, and only negligibly changed the PCr/Pi ratio from starvation.
• 4.4. These results suggest a decrease in creatine kinase activity in the muscle of starved frogs.

     

Aerobic glucose disposal in the oyster: An in vivo kinetic analysis

• 1.1. Aerobic glucose disposal in starved oysters exposed to 1 mM external glucose was 2.29 μg C/g wet wt/min.
• 2.2. It was hypothesized that the maximum disposal rate is limited by the maximum rate of transepithelial glucose transport.
• 3.3. The major recipients of glucose-carbon were glycogen and amino acids. 4. The rate of glucose-carbon disposal to these two pools was 0.80 and 0.42 μg C/g/min, respectively.
• 4.5. The internal energy state determines the pathways of glucose disposal.
• 5.6. Disposal of glucose-carbon in “glucose-primed” oysters is primarily into glycogen.
• 6.7. In fasted bivalves the disposal is primarily into amino acids and carboxylic acids.
• 7.8. The uptake of dissolved glucose has the potential of contributing significantly to growth under conditions where the external glucose concentration is kept artificially high.

     

Determination of β-phenylethylamine in catfish (Parasilurus asotus) tissues by gas chromatography/mass spectrometry

• 1.1. β-Phenylethylamine (PEA) was detected and quantitated in tissues of the catfish, Parasilurus asotus, by very specific and sensitive gas chromatography/mass spectrometry.
• 2.2. The selected ion monitoring was made with a strong quasi-molecular ion of the pentafluoropropionic derivative of PEA in the positive chemical ionization mode.
• 3.3. PEA was found in all tissues tested ranging from 2.8 to 38.2 ng/g wet wt tissue. It was highest in the spinal cord, followed by the skin, brain and intestine.

     

In brief

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The effect of cryopreservation and various cryodiluents on allozyme variation of glucose phosphate isomerase in the F1 progency of african catfish (clarias gariepinus)

• 1.1. African sharptooth catfish individuals, heterozygous for glucose phosphate isomerase (GPI-2), were selected as broodstock by using horizontal starch-gel electrophoresis.
• 2.2. Ova of one heterozygous female were inseminated with cryopreserved and fresh milt from a corresponding male.
• 3.3. Significant deviations from expected Hardy-Weinberg proportions occured for offspring obtained by using cryopreserved milt.
• 4.4. Differences in genotypic variation seems to relate to different cryodiluents and the fertility thereof.
• 5.5. Selection of breeding stock for aquacultural practices based on the above information is discussed.

     

The biochemical and energetic composition of somatic body components of echinoderms from the northern Gulf of Mexico

• 1.1. The biochemical and energetic compositions of the somatic body components of seven species of asteroids, one ophiuroid, and four echinoids from the northern Gulf of Mexico (30–95 m depth) were ascertained.
• 2.2. Levels of ash were high (68.5–90.8% dry wt) in all body-wall tissues, with the exception of the asteroid Echinaster modestus (51.6% dry wt). Levels of ash were low in the pyloric cecae (nutrient storage organ) of asteroids (4.6–30.8% dry wt).
• 3.3. Levels of lipid (8.1–34.5% dry wt), soluble protein (15.9–28.7% dry wt), and insoluble protein (18.1–48.6%, dry wt) were high in the pyloric cecae of all asteroids, but generally low in ophiuroid and echinoid body-wall tissues. High protein levels (28.5–44.5% dry wt) in the body-wall of the asteroids Echinaster modestus and Anthenoides pierceisuggest it may play a role in nutrient storage.
• 4.4. All somatic tissues contained low levels of carbohydrate (0.2–1.4% dry wt).
• 5.5. Levels of energy in pyloric cecal tissues (12.99–26.05 kJ/g dry wt) were 4–8 times higher than in echinoderm body-wall tissues (2.92–11.91 kJ/g dry wt).
• 6.6. The biochemical and energetic compositions of echinoderms from the northern Gulf of Mexico are similar to those of species from other latitudes and depths.

     

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⁺.

     

The influence of anaerobiosis on the levels of adenosine nucleotides and some glycolytic metabolites in tubifex sp. (annelida,oligochaeta)

• 1.1. In Tubifex sp. the amounts of ATP, ADP and AMP, and of glucose, glucose-1-P, glucose-1-P, glucose-6-P, fructose-6-P and fructose-1, 6-P were measured after experimental anaerobiosis.
• 2.2. The energy charge decreased from 0.84 to 0.07/0.69 within 6–9 hr of anaerobiosis.
• 3.3. During long term anaerobiosis there was no change from 0.70/0.69.
• 4.4. The concentrations of glucose, glucose-6-P and fructose-1,6-P increased somewhat during an initial phase of anaerobiosis.
• 5.5. The data are discussed with respect to the regulation of energy metabolism, especially during the transition of aerobic to anaerobic metabolism.
• 6.6. It is concluded that this transition is accomplished within 6–12 hours.