Bioelectrical potentials across the mantle epithelium of Achatina fulica

• 1.1. The shell side of the mantle of Achatina fulica is several millivolts positive to the blood side in vitro.
• 2.2. The electrical potential does not depend on Na⁺, Ca²⁺, Mg²⁺, K⁺ or HCO₃⁻ but requires the presence of chloride on the shell side.
• 3.3. The potential difference and short-circuit current ranged from 3.0 to 30.0 mV and 15.0 to 75 μA/cm² with averages at 10m V and 50 μA/cm² respectively.
• 4.4. The electrical gradient is reduced by 2,4-dinitrophenol, thiocyanate and furosemide but not by ouabain, CO₂ or acetozolamide.
• 5.5. It is suggested that the nature and mechanism of electrogenesis in Achatina parallels that of the Helix mantle.

     

Prostaglandin E1 and analogs of prostacyclin influencing superoxide production by the human neutrophil nadph oxidase system

• 1.1. The effects of prostaglandin (PG) E₁, and I₂ analogs (OP-41483 and OP-2507) on the Superoxide generation of human neutrophil NADPH oxidase (EC 1.6.99.6) in both whole-cell and cell-free systems were investigated.
• 2.2. In a whole-cell system, OP-2507 inhibited the Superoxide generation by neutrophils exposed to phorbol myristate acetate concentration-dependently through its superoxide-scavenging action.
• 3.3. The concentration of the drug required for 50% inhibition of the oxidase (IC₅₀) was 21 μM.
• 4.4. In a cell-free system, however, the drug in concentrations of < 100 μM did not inhibit the activation of NADPH oxidase by sodium dodecyl sulfate because of its inactivation by the detergent.
• 5.5. Although PGE₁ and OP-41483 did not inhibit the Superoxide production by stimulated neutrophils in a whole-cell system, they both inhibited the activation of NADPH oxidase in a cell-free system concentration-dependently, with IC₅₀ values of 44 and 170 μM, respectively.
• 6.6. In addition, in the cell-free system, the K_m value for NADPH of the oxidase was unchanged by PGE₁.
• 7.7. The results suggest that the PGI₂ analog, OP-2507, is a possible superoxide-scavenger and that PGE₁ inhibits the NADPH oxidase activation by sodium dodecyl sulfate in a cell-free system concentration-dependently.

     

Ostrich fructose-1,6-bisphosphatase: Kinetic characterization of the liver isoenzyme

• 1.1. Purified ostrich (Struthio camelus) liver fructose-1,6-bisphosphatase exhibited an absolute requirement for Mg²⁺.
• 2.2. The enzyme catalyzed the hydrolysis of fructose-1,6-bisphosphate, sedoheptulose-l,7-bisphosphate and ribulose-l,5-bisphosphate.
• 3.3. S_0.5 for substrate was 1.4 μM.
• 4.4. AMP was a potent non-competitive inhibitor with respect to substrate (K_i of 25 μM).
• 5.5. Fructose-2,6-bisphosphate was a potent competitive inhibitor of the enzyme (K_i of 4.8 μM).

     

Aerobic and anaerobic respiration of adult Artemia salina L., acclimated to different oxygen concentrations

• 1.1. Adult male Atremia salina L. were acclimated to five different oxygen concentrations and their respiration in response to environmental oxygen concentrations was determined.
• 2.2. Anemia is a respiratory regulator over a wide range of partial O₂ pressures. A critical oxygen tension exists and decreases with acclimation to lower pO₂.
• 3.3. Hypoxic conditions induce the production of hemoglobin III.
• 4.4. Lactic acid is produced during anaerobiosis.
• 5.5. Production of Hb III and lactic acid, being inversely proportional to the acclimation level, has to be considered as a long term or short term adaptation to hypoxic conditions.

     

The effect of d2o on glycolysis by rat hepatocytes

• 1.1. The effect of incorporating D₂O into the incubation medium on glycolysis and gluconeogenesis by hepatocytes from fasted rats was examined.
• 2.2. The substitution by heavy water, D₂O, at concentrations from 10 to 40%, stimulated glucose uptake, lactate production and CO₂ yields from glucose. At 10 mM glucose, 40% D₂O doubled glucose uptake, increased CO₂ production by 40%, and increased lactate production by 350%.
• 3.3. The stimulation of lactate production decreased at higher glucose concentrations, but was still substantial even at 80 mM glucose.
• 4.4. There was no effect on CO₂ production above glucose concentrations of 30 mM.
• 5.5. Ten percent D₂O showed little inhibition of lactate uptake, its oxidation and gluconeogenesis. At 40% D₂O the inhibition ranged from 10 to 20%.
• 6.6. No effect of D₂O on the rate of glucokinase or glucose-6-phosphatase was observed.
• 7.7. The concentration of fructose, 2,6-P was not affected by D₂O

     

Partial purification of rat liver cytoplasmic acetyl-CoA synthetase; Characterization of some properties

• 1.1. Rat liver cytoplasmic acetyl-CoA synthetase was partially purified (purification factor = 23, yield = 30%).
• 2.2. The apparent K_ms for acetate, coenzyme A, ATP and MgCl₂ were determined and found to be 52.5 μM, 50.5 μM, 570 μM and 1.5 mM, respectively.
• 3.3. The partially-purified enzyme showed a low affinity for short-chain carbon substrates other than acetate.
• 4.4. The properties of the partially-purified enzyme were compared with those of enzymes from other sources.

     

Locality-dependent concentrations of bromophenols in Lanice conchilega (Polychaeta: Terebellidae)

• 1.1. At 37 stations in the English Channel and the southern North Sea concentrations of four brominated phenols in Lanice conchilega were determined using ECD equipped capillary gas chromatography: 2,4-dibromophenol, 2,6-dibromo-4-methylphenol, 2,4,6-tribromophenol, 3,5-dibromo-4-hydroxybenzaldehyde.
• 2.2. Concentrations in worms of the southern North Sea were generally below 1 μg/g wet wt.
• 3.3. Levels were slightly raised in worms of sheltered shores, those of 3,5-dibromo-4-hydroxybenzaldehyde were increased in subtidal populations.
• 4.4. Concentrations in L. conchilega of Brittany were considerably higher than those in worms of the Frisian Islands, North Sea; they deviated from the latter by factors of 3, 16, 4, 4, respectively.
• 5.5. The reasons for the conspicuous differences are hitherto unknown; three explanations are suggested.

     

Some aspects of activation and inhibition of rat brain lipoxygenase

• 1.1. Regulatory properties of lipoxygenase activity in rat brain cytosol were studied using linoleic acid (LA) as a substrate.
• 2.2. A change in the absorbance at 234 nm was biphasic when a mixture of LA and pre-formed hydroperoxide (LA-OOH) was incubated with freshly isolated native brain cytosol. Initially, a rapid depletion of LA-OOH was observed with a concomitant formation of LA-oxo compounds. This phase was followed by LA dioxygenation.
• 3.3. Both hydroperoxidase and dioxygenase activities of lipoxygenase were inhibited by micromolar concentrations of classic lipoxygenase inhibitors (phenidone, 5,8,11-eicosatriynoic acid and nordihydroguaiaretic acid).
• 4.4. The dioxygenase activity in dialysed cytsool was stimulated by nanomolar concentrations of H₂O₂ and micromolar concentrations of LA-OOH and it was inhibited by serotonin, dopamine and norepinephrine (IC₅₀ 25–43 μM).

     

Characterization and purification of biliverdin reductase from the liver of eel,Anguilla japonica

• 1.1. Biliverdin reductase from the liver of eel, Anguilla japonica was characterized and purified with a novel enzymatic staining method on polyacrylamide electrophoretic gel.
• 2.2. This enzyme could use both NADPH and NADH as coenzyme. The K_m of NADPH was 5.2 μM, while that of NADH was 5.50 μM.
• 3.3. The optimum reaction pH for using HADPH as coenzyme was 5.3. That for NADH was 6.1. The optimum reaction temperature is 37°C.
• 4.4. When NADPH was used as coenzyme, the K_m of biliverdin was 0.6 μM. When NADH was used as coenzyme, the K_m of biliverdin was 7.0 μM.
• 5.5. The activity of the enzyme was inhibited by the concentration of biliverdin. Also, the potency of the enzyme was much less than that of the analogous enzyme isolated from mammals.
• 6.6. This is a fairly stable enzyme with a mol. wt around 67,000. Its estimated pI was pH 3.5–4.0.
• 7.7. This is the first time biliverdin reductase has been isolated and characterized from a vertebrate other than mammals. The property of it is quite different from that of mammals.

     

Kinetic analysis of rat liver sorbitol dehydrogenase

• 1.1. A steady state kinetic investigation was performed on an improved preparation of rat-liver sorbitol dehydrogenase (l-iditol: NAD-oxidoreductase, EC 1.1.1.14).
• 2.2. Data analyses indicate the enzyme follows a rapid equilibrium random mechanism in the direction of sorbitol oxidation and a random mechanism in the direction of fructose reduction.
• 3.3. Kinetic constants were: K_m^NAD 0.082 mM; K_m^sorbitol 0.38 mM; K_m^NADH 67 μm; K_m^fructose 136 μM.
• 4.4. Evidence is adduced to indicate the more rapid reverse (fructose reduction) reaction is susceptible to metabolic control by formation of abortive enzyme-fructose-NAD and enzyme-NADH-sorbitol complexes.

     

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.

     

Purification and characterization of a blue-colored red-fluorescent protein from the silkworm (Bombyx mori L.) larvae

• 1.1. A red-fluorescent blue protein (P600) was purified from the digestive juice of the silkworm (Bombyx mori L.) larvae raised on mulberry leaves.
• 2.2. The purified protein was electrophoretically homogeneous and showed the absorption maxima at 601.5 nm and 278 nm, and the fluorescence maximum at 621 nm.
• 3.3. The molecular weight was estimated to be 540,000 by gel filtration on Sepharose CL-6B. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate suggests that the protein consists of two heterogeneous polypeptide subunits with a mol. wt of 15,000 and 18,000.
• 4.4. The P600 contains excess acidic amino acid residues over basic groups. The polarity and pI were 45.5% and 4.6, respectively.
• 5.5. The production of H₂O₂ was observed in the presence of P600 upon illumination.

     

Purification and some properties of an atypical alkaline p-nitrophenylphosphate phosphatase activity from Halobacterium halobium

• 1.1. An alkaline p-nitrophenylphosphate phosphatase has been purified 440-fold from extracts of Hatobacterium halobium.
• 2.2. The enzyme has an apparent molecular weight of 24,000.
• 3.3. A K_m value for p-nitrophenylphosphate of 1.12mM has been found under optimal conditions.
• 4.4. The enzyme is selectively activated and stabilized by Mn²⁺.
• 5.5. It requires high salt concentrations for stability and maximum activity.
• 6.6. It displays an unusual restricted substrate specificity of 25 phosphate esters tested, only phosphotyrosine and casein were hydrolysed besides p-nitrophenylphosphate.

     

An alkaline phosphatase from Thermus sp strain Rt41A

• 1.1. A thermostable orthophosphoric monoester phosphohydrolase (EC 3.1.3.1) from Thermus sp strain Rt41A has been purified 400-fold to give a specific activity of 25 U/mg at 60°C in IM diethanolamine (pH 11.1).
• 2.2. The enzyme has a M_r of 160,000 and is trimeric.
• 3.3. The half-life of the enzyme is 5 min at 85°C.
• 4.4. The enzyme has a wide specificity for a number of phosphate monoesters.
• 5.5. The H_m of the enzyme is pH dependent, so the pH optimum of the enzyme is affected by the substrate concentration.
• 6.6. The enzyme is inhibited 50% by 20 mM Ca²⁺ or Mg²⁺.
• 7.7. The K_i for phosphate, EDTA-di sodium salt and arsenate (in 1 M diethanolamine, pH 11.1) is approx 1.2, 1.6 and 4mM respectively.
• 8.8. Urea (200 mM) is not inhibitory.

     

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.

     

Decarboxylation of uroporphyrinogen I and III in 2,3,7,8-tetrachlorodibenzo-p-dioxin induced porphyria in mice

• 1.1. The decarboxylation of uroporphyrinogens I and III by porphyrinogen carboxy-lyase (EC 4.1.1.37) in mouse liver supernatant was compared in relation to substrate concentrations.
• 2.2. In this species uroporphyrinogen III was the best substrate judging by the criteria of K_m/V_max (estimated for total porphyrinogens) and was converted into coproporphyrinogen faster than its series I isomer.
• 3.3. The difference between the two isomers was mainly due to the first decarboxylation.
• 4.4. This difference was confirmed by calculation of the Hill coefficient and of Lineweaver-Burk plot which suggested that isomer I induced negative cooperativity in the active centre of the enzyme.
• 5.5. After treatment with a porphyrogenic dose of TCDD (25 μg/kg/week for 9 weeks) differences between uroporphyrinogen I and III as substrate were maintained.
• 6.6. In addition treatment reduced V_max and K_m (estimated for total porphyrinogens) of liver porphyrinogen carboxy-lyase to about half control values for both isomers.
• 7.7. V_max was reduced mainly because of the formation of smaller amounts of all products of decarboxylation, and K_m because more heptaporphyrinogen was formed than coproporphyrinogen.
• 8.8. Values of the Hill coefficient and Lineweaver-Burk plots suggested TCDD induced altered substrate affinity for isomer III too.
• 9.9. Treatment with TCDD did not affect the decarboxylation of uroporphyrinogen III by RBC porphyrinogen carboxy-lyase, estimated from K_m and V_max for total porphyrinogens formed.

     

A note on eggshell porosity,nest humidity and the effects of DDE in the grey heron (Ardea cinerea)

• 1.1. Water vapour conductance (G_H2O) was determined for 25 grey heron Ardea cinerea eggs in the laboratory, and in nests during natural incubation at two Scottish colonies.
• 2.2. The mean G_H2O of eggs measured in the nest which successfully hatched was 9.0 mgH;O/mmHg/day and the mean water vapour pressure gradient between egg and nest (ΔP_H2O), measured using “calibrated” duck eggs, averaged at 31 mmHg (4.13 kPa).
• 3.3. Based on eggshell porosity results, from the eggs which hatched, such a gradient would result in a loss of water from the eggs during incubation equivalent to 11% of their fresh weight.
• 4.4. Shell thickness, the number of pores/cm² of eggshell and DDE content were also determined for the 25 eggs measured in the laboratory.
• 5.5. Eggs containing high levels of DDE had thinner shells, more pores in the eggshell and a higher overall eggshell porosity.
• 6.6. The main problem posed by a high level of DDE would appear, however, not to be an excessive water loss from the egg during incubation, but rather eggshell thinning leading to a loss of the egg due to breakage in the nest.

     

Cellular and intracellular localization of catalase and acid phosphatase in the midgut of Lumbricus terrestris L.: A cell fractionation study

• 1.1. Cellular and intracellular localization of catalase and acid phosphomonoesterase in the midgut of Lumbricus terrestris was studied by use of tissue fractionation.
• 2.2. At least 60–70% of the catalase resides in the chloragocyte cytosol and the remaining 30–40% resides in gut epithelium peroxisomes.
• 3.3. One of the main functions of the chloragocyte catalase is probably scavenging for H₂O₂ arising from the interaction between blood heme-protein and oxygen.
• 4.4. A simple method for the histochemical detection of cytosol catalase is proposed.
• 5.5. About 10% of the gut acid phosphatase resides in chloragocyte lysosomes. The chloragosomes contain no acid phosphatase.

     

Time related changes in the free amino acid pool of the sea anemone,Bunodosoma cavernata,during salinity stress

• 1.1. The sea anemone, Bunodosoma cavernata, is a relatively eurybaline cnidarian tolerating salinities from 12 to 40%.
• 2.2. Taurine, glutamic acid and aspartic acid all showed some increases with increased salinity.
• 3.3. The amino acid showing the greatest accumulation under high salinity conditions was β-alanine which increased 28-fold from 1.5 to 41.9 μmol/g dry weight when salinity was raised from 26 to 40%.
• 4.4. When B. cavernata was subjected to increased salinity, β-alanine was rapidly accumulated and reached maximum levels within 4 days.
• 5.5. When salinity was dropped from 36 to 26%0, β-alanine concentrations dropped from 15 to 2 μmol/g dry weight in 2 days.

     

The oxygen consumption rates of three gastrotrichs

• 1.1. The oxygen consumption rates for three sympatric species of marine gastrotrichs (anatomically similar, except that one contains hemoglobin) were measured with a Cartesian diver microrespirometer.
• 2.2. The rates for the two species without hemoglobin, Turbanella ocellata and Dolichodasys carolinensis, were 307.2 μl O₂ g⁻¹ hr⁻¹ and 108.0 μl O₂ g⁻¹ hr⁻¹, respectively, while the rate for the hemoglobin-containing species, Neodasys, was 208.9 μl O₂ g⁻¹ hr⁻¹.
• 3.3. The possession of hemoglobin by Neodasys (14% by volume) cannot be explained by an unusually high demand for oxygen.
• 4.4. Instead, the hemoglobin may be useful as an oxygen store providing continued aerobic metabolism in anoxic conditions, thus allowing Neodasys to exploit a different niche.