Mitochondria from the ventricle of the marine clam,Mercenaria mercenaria: Substrate preferences and effects of pH and salt concentration on proline oxidation

• 1.1. Mitochondria with high respiratory control ratios (RCR) have been isolated from the ventricle of the marine clam Mercenaria mercenaria.
• 2.2. Proline is the preferred substrate of the mitochondria of the ventricle based on state 3 rates.
• 3.3. Pyruvate, ornithine and succinate are oxidized at rates 3/4 that of proline.
• 4.4. α-Glycerophosphate was oxidized at rates 1/2 that of proline.
• 5.5. The pH optimum for proline oxidation lies between 6.5 and 7.5 based on RCR and ADP/O and between 7.0 and 7.4 based on state 3 rates.
• 6.6. KCl concentrations between 250 and 450 mM gave optimal values for the oxidation of proline based on RCR and state 3 rates.
• 7.7. KCl concentration had little effect on ADP/O between 100 and 850 mM.

     

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.

     

Interrelationships between gluconeogenesis and uricogenesis in chicken liver

• 1.1. Experiments performed on isolated hepatocytes and perfused liver of starved chickens showed that gluconeogenesis from lactate, glycerol and fructose was inhibited by 22–100% on addition of urate precursors.
• 2.2. The inhibition was associated with an increased rate of urate formation.
• 3.3. 2,4-Dinitrophenol (40 μM), 2-bromooctanoate (2 mM) and 3-mercaptopicolinate (3MPA) (0.5 mM) were inhibitory with respect to gluconeogenesis but did not significantly affect the rate of urate formation.
• 4.4. The possible interrelationships between gluconeogenesis and uricogenesis are considered in terms of a competition for ATP and for other metabolites between the two pathways.
• 5.5. An interplay of both pathways at the level of anion transfer across the inner mitochondrial membrane is also discussed.

     

Specificity of the permissive effect of d-Glucose on insulin release in chicken pancreas

• 1.1. As previously shown, 14 mM d-glucose, a non-insulinotropic concentration in isolated chicken pancreas, permits an insulin release in response to d-glyceraldehyde, (d-GA; a glycolytic fuel) and l-leucine or α-ketoisocaproic acid (α-KIC) (non-glycolytic fuels), which alone are not initiators of insulin release in this species.
• 2.2. The “permissive” effect of d-glucose was also observed in the presence of d-mannose (which, as shown herein, is not insulinotropic alone).
• 3.3. The specificity of glucose for this “permissive” effect was, therefore, subsequently questioned in the presence of 10mM α-KIC by substituting various glycolytic and non-glycolytic fuels to glucose.
• 4.4. d-GA (at 5 and 15mM), d-mannose (30 and 50 mM), or the association of l-glutamine + l-asparagine permitted an insulin release in response to α-KIC.
• 5.5. The response was, however, delayed with d-GA, only occasionally with 50 mM d-mannose, and required high concentrations and was delayed in the presence of l-glutamine + l-asparagine as compared to that obtained with 14mM d-glucose + α-KIC.
• 6.6. In conclusion, the threshold of fuel-induced insulin release is much higher in the chicken than in mammals and this threshold is most efficiently lowered by glucose.

     

Interaction of lipogenic substrates in porcine adipose tissue in vitro

• 1.1. Porcine adipose tissue was incubated with radiolabeled glucose, acetate or lactate. Saturation curves indicated that lactate > glucose > acetate in providing two-carbon units for fatty-acid synthesis.
• 2.2. Competition between individual substrates indicated that lactate was the best lipogenic substrate.
• 3.3. Incubation of all three substrates at concentrations observable in serum indicated that at 5.56mM, glucose was the preferred lipogenic substrate in the presence of 0.1 mM acetate and 1.0 mM lactate.
• 4.4. At elevated concentrations (18.52mM glucose, 1.0 mM acetate and 10.0 mM lactate), acetate and lactate were preferred to glucose as lipogenic substrates.

     

Relationship between ecg,eeg and sps responses during arousal in the goldfish (Carassius auratus)

• 1.1. Goldfish (Carassius auratus) were fitted with ECG electrodes and intra-cranial stainless steel electrodes to monitor cardiac, EEG and SPS responses, during minimal restraint, to presentation of environmental stimuli (light-on, moving shadow, tap).
• 2.2. All three stimuli evoked a bradycardia and increases in the EEG frequency; correlates of arousal responses in fish.
• 3.3. EEG frequency changes were most evident in the fore- and midbrain regions; in the hindbrain smaller responses nevertheless showed discrimination between stimuli.
• 4.4. EEG amplitude changes were more site- and stimulus-specific than frequency changes.
• 5.5. SPSs occurred on stimulus presentations which were negative in polarity in the midbrain and positive in the forebrain and hindbrain, though the latter were smaller and less consistent.
• 6.6. Principal components analyses and regression analyses were used to examine detailed associations between peripheral and central physiological changes.
• 7.7. It was found that increases in the EEG frequency of fore- and midbrain regions were related to cardiac deceleration on early stimulus presentations.
• 8.8. This was also shown for the negative SPS of the midbrain to the presentation of the tap stimulus.
• 9.9. Positive SPSs of the forebrain were related to the bradycardia on later stimulus presentation i.e. during habituation of the arousal response.
• 10.10. The results indicate that in fish, as in other vertebrates, negative SPSs in the midbrain are associated with arousal and implicate the forebrain in the modulation of arousal by its habituation.

     

Serotonin or electrical optic nerve stimulation increases the photosensitivity of the Aplysia eye

• 1.1. Exposure of isolated Aplysia eyes to serotonin (10⁻⁷ M) produces large and long-lasting (hours) increases in the ERG recorded from the surface of the eye.
• 2.2. Dopamine, octopamine, or acetylcholine do not mimic the effect of 5-HT on the ERG.
• 3.3. Brief electrical optic nerve stimulation (2 Hz, 2 min) also increases the ERG and this effect also lasts a long period of time (0.5–2 hr).
• 4.4. Our results suggest that serotonin increases the response of photoreceptor cells to light and that efferent optic nerve activity may modulate photosensitivity through release of serotonin in the eye.

     

Characteristics of Crassostrea virginica crystalline style chitin digestion

• 1.1. Fundamental chitin digestion characteristics of Crassostrea virginica crystalline style were investigated.
• 2.2. Optimum temperature and pH were 34°C and 4.8. respectively.
• 3.3. The colloidal regenerated chitin (0.56mol/0.5 ml: GlcNAc equivalents) was saturating under all enzyme levels encountered.
• 4.4. There was no evidence of end product inhibition, even after 100 hr incubation.
• 5.5. Calculated K_m for the chitinase complex was 1.19mM when determined using a 30 min assay, but was only 0.70 mM when determined using a 4.6 hr assay.
• 6.6. Both K_m values are lower than reported for similar assays in other molluscs and for most bacteria.
• 7.7. Effect of substrate preparation on the kinetics are discussed.
• 8.8. Eight peaks of chitinase activity were resolved by DEAE-Fractogel ion exchange chromatography.

     

A study of excitatory efferent fibres in the intestinal nerve of the fowl (Gallus domesticus)

• 1.1. The intestinal nerve of the fowl was studied in vitro.
• 2.2. A significantly larger amplitude spike discharge was recorded in side branches of the nerve which innervate the gut when the aboral end of the main nerve trunk was stimulated than when the oral end was stimulated.
• 3.3. Postganglionic autonomic neurones innervating the smooth muscle of the ileum are not located in the intestinal nerve. Evidence is presented, however, supporting the idea that such neurones innervating the rectum are located in the rectal position of the nerve.
• 4.4. The increase in intraluminal pressure and circular muscle tension in the ileum was greater following aboral nerve stimulation than following oral nerve stimulation.
• 5.5. It is suggested that excitatory efferent nerve fibres ascend the intestinal nerve to innervate the ileum.

     

Properties of the transition from short-term to long-term depression in neurons of Helix pomatia

• 1.1. Synaptic short-term depression could be transferred into long term depression by repetition of series of stimuli.
• 2.2. The transition from short-term depression to long-term depression was blocked by puromycin.
• 3.3. The majority of the transition took place during resting periods between stimulus series.
• 4.4. The initiation of the transition process was 83% completed after 5 min of stimulation.
• 5.5. Short- and long-term depression were quantitatively separated into their two serial sites of origin: afferent axons and synaptic terminals.
• 6.6. Long sequences evoked periods with increased and variable EPSPs not conforming to depression.

     

The sea urchin extraembryonic hyaline layer: Ionic parameters controlling the hyalin gelation reaction

• 1.1. As reported previously (Robinson, 1988) the Ca²⁺-induced self-association reaction of the protein hyalin, purified from the sea urchin extraembryonic hyaline layer, was modulated by both Mg²⁺ and NaCl.
• 2.2. In the presence of 400 mM NaCl the apparent dissociation constant (Ca²⁺) decreased five-fold from 4.8 ± 1.1 mM in the absence to 0.9 ± 0.5 mM in the presence of 20 mM Mg²⁺.
• 3.3. The potentiating effect of Mg²⁺ occurred with an apparent dissociation constant (Mg²⁺) of 4.6 ± 0.5mM.
• 4.4. In the absence of Ca²⁺ or NaCl hyalin dissociated from isolated hyaline layers indicating that the behavior of hyalin within the layer is predictable from results obtained with the purified protein.

     

Functional differentiation of crayfish statocyst receptors in sensory adaptation

• 1.1. Response characteristics of tonic- and phasic-type receptors in the crayfish statocyst were investigated with intracellular recording technique.
• 2.2. They were identified to be either tonic- or phasic-type according to their response patterns to the hair deflection performed by water jet stimulation.
• 3.3. Constant depolarizing current applied intracellularly evoked long-lasting spike discharge in the tonic-type neurons and transient discharge in the phasic-type neurons.
• 4.4. These tonic- and phasic-type neurons also showed different patterns of spike discharge to depolarizing pulse stimulus of 50 msec duration.
• 5.5. On the basis of the response patterns to this pulse stimulus, it was shown that the statocyst receptor neurons consist of 46% tonic- and 54% phasic-type neurons.

     

Effects of l-canavanine on arginine catabolism in manduca sexta (sphingidae: lepidoptera)

• 1.1. Hemolymph ornithine concentrations in tobacco horn worm larvae fed a 2.5 mM l-canavanine plus 25 mM l-arginine-supplemented artificial diet (CAAM) were higher than those in larvae fed diets supplemented with 2.5 mM canavanine (CAV), 25 mM arginine (ARG), or controls (CON).
• 2.2. Ornithine concentrations in CAV-treated larvae were significantly greater than the control or ARG treatment, but less than the CAAM treatment during the latter part of the wandering larval stage and during the pharate pupal stage.
• 3.3. Urea concentrations were greater during the active feeding stage with the CAAM- and ARG-treated larvae having significantly higher levels than control or CAV-treated larvae.
• 4.4. Urea concentrations in all treatments never exceeded 36.5% of the ornithine concentration.
• 5.5. Canavanine concentrations were higher in CAV-treated larvae than in CAAM-treated larvae.
• 6.6. During active feeding, arginine concentrations for all treatments were similar, but were lower in CAV- and CAAM-treated larvae during the pharate pupal stage.

     

Evidence for hyporegulation in the calanoid copepod,Acartia tonsa

• 1.1. The euryaline calanoid copepod, Acartia tonsa, maintains haemolymph Na below that of the external medium in salinities above 34^o_oo (475 mM Na).
• 2.2. The measured transepithelial electrical potential. −9.97 ± 1.0 mV, indicates that Na is regulated out of electrochemical equilibrium.
• 3.3. Water osmotically lost in hyporegulation is replaced by Na-dependent absorption by the gut.
• 4.4. High osmotic water permeability is evidenced by the fact that with an increase in external salinity from 475 mM Na to 580 mM Na the copepod's drinking rate nearly doubles.
• 5.5. Sodium efflux measurements indicate that ionic permeability is much lower than other hyporegulating crustaceans.
• 6.6. The energetic advantage of hyporegulation in this species is considered.

     

Properties and distribution of Mg2+-HCO−3-ATPase in brush border membranes isolated from rat small intestine

• 1.1. The small intestine was cut into seven segments and properties and distribution of brush border Mg²⁺-HCO⁻₃-ATPase activity in each segment were examined.
• 2.2. The optimal Mg²⁺ concentration was 1.0 mM.
• 3.3. The optimal HCO⁻₃ concentration was 100 mM in the first (duodenal), 50 mM in the 3rd and 40 mM in the 5th segment, respectively.
• 4.4. The optimal pH value was about 9.0.
• 5.5. l-phenylalanine (above 1 mM) and SCN⁻ (above 50 mM) significantly inhibited both Mg²⁺- and Mg²⁺-HCO⁻₃-ATPase activity.
• 6.6. The enzyme activity was found to be highest in the duodenal segment and then gradually decreased in consecutive segments as well as β-glycerophosphatase, Na⁺-K⁺-ATPase and supernatant carbonic anhydrase.
• 7.7. The functional significance of this ATPase and the relationship with carbonic anhydrase was discussed.

     

Axonal mapping of neurosecretory Helix bursting cells. Functional aspects of peripheral multibranched axons

• 1.1. The peripheral axon distribution from two bursting neurons was investigated using electrophysiological methods. Both of these cells send out a bundle of axon branches which goes via the visceral nerve to the heart and in the uterine nerve.
• 2.2. The relative number of axon branches in the two nerves was determined through double-shock experiments.
• 3.3. The axon bundle takes the visceral nerve and its uterine branch, supplying the effector systems in parallel.
• 4.4. Slight differences in conduction velocity between the axon branches produce an enlargement of the efferent volley.
• 5.5. The number of active axon branches conveying orthodromic or antidromic spikes is controlled by inhibitory potentials converging onto the initial part of the bundle, so that the two bursting cells amount to a pool of 35 to 40 interconnected nerve cells.
• 6.6. The atypical axonal distribution of the two bursting cells might be related to their neurosecretory properties.

     

A comparative study of nuclear and chloroplast DNA dependent RNA polymerases from wheat leaves

• 1.1. Three DNA dependent RNA polymerases have been purified from chromatin and chloroplast fractions of wheat leaves.
• 2.2. The purified enzymes were completely dependent on exogenous DNA after purification by glycerol gradient, DEAE-Sephadex and phosphocellulose chromatography.
• 3.3. The nuclear enzymes, I and II, showed a strong preference for denatured nuclear DNA, whereas the chloroplast enzyme preferred denatured chloroplast DNA.
• 4.4. The three enzymes require either Mg²⁺ or Mn²⁺ for activity.
• 5.5. α-amanitin specifically inhibited RNA polymerase II but has no effect on polymerase I and chloroplast polymerase.
• 6.6. Enzyme I is most active at very low ionic strength (0.10 mM KC1), whereas enzyme II and chloroplast enzyme show maximum activity at 150mM and 50 mM KC1 respectively.

     

Beef liver glutamate dehydrogenase : Effects of partial proteolysis with chymotrypsin

• 1.1. After chymotryptic digestion of bovine glutamate dehydrogenase, the assay conditions determine whether activation or inhibition is observed.
• 2.2. The major fragments appear to remain physically associated.
• 3.3. Responses to both GTP and ADP are altered. Inhibition by GTP at pH 7 and 8 is almost abolished.
• 4.4. Out of various ligand combinations tested, GTP and NADH together provide the best protection against all the proteolytic effects.

     

Disruption of endothelial barrier function: relationship to fluidity of membrane extracellular lamella

• 1.1. Endothelial cells were cultured in tissue culture flasks or on microcarrier beads and labeled with a lipid specific spin-label.
• 2.2. Exposure of endothelial cells to benzyl alcohol caused a dose- and time-dependent increase in membrane fluidity using electron spin resonance (ESR). Maximum fluidity was reached after a 5-min exposure to 100 mM benzyl alcohol.
• 3.3. Albumin permeability across endothelial cells cultured on micropore filters was used as an indication of endothelial monolayer integrity.
• 4.4. A significant increase in permeability occurred with 50 mM benzyl alcohol. Maximal albumin permeability was reached after a 5-min exposure to 100 mM benzyl alcohol.

     

Activation of neutrophils NADPH oxidase by PMA: Cytosol activity is translocated in phorbol-primed neutrophils

• 1.1. Translocation of cytosol activity in phorbol-primed neutrophils was studied.
• 2.2. Prior exposure of PMA or FMLP could potentiate the oxidative response by subsequent heterogeneous stimulus, FMLP or PMA.
• 3.3. In FMLP-primed neutrophils, the cytosol had almost the same activity as resting one and cytosol activity was not eluted from the membrane.
• 4.4. In PMA-primed neutrophils, however, the cytosol had less activity and cytosol activity was correspondingly eluted from the membrane.
• 5.5. These observations suggested that cytosol activity was translocated in PMA-primed cells.