Intracellular proteolytic activity in developing myxospores of Myxococcus xanthus

1. Cell-free extracts from vegetative cells and developing myxospores of Myxococcus xanthus were found to contain similar amounts of proteolytic activity, approximately 80% of which was due to one or more neutral metal proteases.
2. Sixty per cent of the proteolytic activity was particulate.
3. The specific activity of the proteases was high throughout all stages of myxospore formation and displayed small increases in activity at two stages of development: (1) during cell shortening and (2) immediately following the conversion to spheres. The first peak in activity was apparent in assays conducted at pH 8 or 10 whereas the second peak was obvious only at pH 6.
4. A mutant which develops into myxospores only after a lag of approximately 7–8 h possessed levels of proteases similar to the wild type and displayed a peak in proteolytic activity after a delay of 7–8 h.
5. Low levels of serine protease activity were occasionally detected in both vegetative cells and myxospores; no sulfhydryl proteases were detectable in either cell type.
6. Extracellular proteases accumulated in the medium throughout myxospore development but differed from the intracellular proteases in pH optima and sensitivity to inhibitors.

     

Seasonal variation in the biochemical composition of Mytilus viridis at Ratnagiri on the West Coast of India

1. The seasonal variation in the water, protein, fat and glycogen contents of the mussel, Mytilus viridis has been studied for the year March, 1974 to March, 1975.
2. The water level increased during the monsoon season and decreased in summer.
3. The level of protein, fat and glycogen showed correlation with the reproductive cycle of the mussel.
4. The protein level was high when the mussels were mature and dropped during the breeding period.
5. During sex change from male to female in May the protein level remained high whereas during sex change from female to male in October and November it was low.
6. The fat level was high in mature mussels and declined on spawning.
7. The glycogen level was at its peak in immature mussels and low in mature.

     

Intracellular calcium in the cells of the outer mantle epithelium of Anodonta cygnea

• 1.1. The intracellular concentration of ionized calcium was measured with double-barrelled ion-sensitive microelectrodes under short-circuit conditions in the isolated outer mantle epithelium of Anodonta cygnea.
• 2.2. When the outside baths contained 1 mmol/1 Ca²⁺ the average intracellular Ca²⁺ was 5.42 ± 0.64 mmol/1(N = 41) while the equilibrium concentration estimated from the intracellular potential measured in the same cells was 5.51 ± 0.33 mmol/l.
• 3.3. Bilateral removal of calcium from the external baths induced a fast fall in the intracellular concentration of this ion by almost three orders of magnitude. This effect was similar to that obtained by removing calcium from the bath on the basolateral side.
• 4.4. Removal of calcium from the bath in contact with the apical side of the preparation had little effect on intracellular calcium.

     

Metabolic products of microorganisms

1. The total nitrogen content of spores of Penicillium notatum increased during swelling and reached a maximum before germ-tubes were formed. It subsequently decreased during germ-tube formation until a constant value in hyphae was reached.
2. An increase in the protein content of the spores was found during swelling, followed by a decrease when germ-tubes were formed. After germination, the protein content increased again to a constant level in hyphae.
3. The content of total lipids steadily decreased during swelling of spores. It reached a minimum value in germinating spores, followed by a net accumulation during hyphal growth. Similar changes occurred in free lipids, phospholipids and the acetone-soluble lipids but not in bound lipids.
4. After an initial decrease during swelling, no further change took place in the neutral carbohydrates content of the spores at the time of germ-tube formation. An accumulation of neutral carbohydrates occurred during late hyphal extension.
5. The nucleic acid content increased sharply during swelling, and reached the highest value in swollen spores just prior to the initiation of germ-tube formation. Afterwards, its content steadily decreased during germ-tube formation and hyphae elongation.

     

Effects of in vivo ethanol administration on Ca2+/Mg2+ ATPase and ATP-dependent Ca2+ uptake activity in synaptosomal membranes

1. Acute administration of ethanol (4 g/kg, i.p.) to mice inhibits the sequestration of calcium into endoplasmic reticulum-like organelles in synaptosomal membranes.
2. Ethanol administration inhibits both Ca²⁺-stimulated adenosine triphosphate hydrolysis and ATP-dependent calcium uptake in the vesicles at time of loss of righting reflex.
3. At recovery of righting reflex, the Ca²⁺-ATPase activity returns to normal levels, while the ATP-dependent uptake remains inhibited.
4. The effect of ethanol is specific for the sequestration (active transport) of calcium since calcium binding to synaptic membranes is not altered.
5. Alteration in mechanisms responsible for synaptosomal buffering of cytosolic Ca²⁺ levels by in vivo ethanol may contribute to altered transmitter release rates following ethanol adminstration.

     

Growth of a floating aquatic weed,Salvinia under standard conditions

1. Growth of the floating aquatic weed, Salvinia, in sterile culture was exponential for at least 2 weeks under standardized conditions.
2. Increase in light intensity or in CO₂ resulted in increases in growth rate, but did not extend the exponential period of growth.
3. This aquatic plant, like many others, discriminates against calcium relative to strontium.
4. In culture Salvinia exhibited luxury consumption of N and P.
5. Because of high C/N ratios, Salvinia may not be a favorable source of animal food, but might be useful in nutrient removal schemes.
6. In sterile culture, S. molesta produced fewer leaves than S. minima, but maintained a significant increase in leaf area and dry weight. This may be correlated with the ability of the first species to rapidly spread over tropical waterways.

     

Selective phonotaxis to advertisement calls in the gray treefrog Hyla versicolor: behavioral experiments and neurophysiological correlates

1. The significance of particular acoustic properties of advertisement calls for selective phonotaxis by the gray treefrog, Hyla versicolor (= HV), was studied behaviorally and neurophysiologically. Most stimuli were played back at 85 dB SPL, a level typically measured at 1–2 m from a calling male.
2. Females preferred stimuli with conspecific pulse shapes at 20° and 24°C, but not at 16°C. Tests with normal and time-reversed pulses indicated the preferences were not influenced by the minor differences in the long-term spectra of pulses of different shape.
3. Pulse shape and rate had synergistic or antagonistic effects on female preferences depending on whether the values of one or both of these properties in alternative stimuli were typical of those in HV or heterospecific (H. chrysoscelis = HC) calls.
4. More auditory neurons in the torus semicircularis were temporally selective to synthetic calls (90%) than to sinusoidally AM tones and noise (< 70%).
5. Band-pass neurons were tuned to AM rates of 15–60 Hz. Neurons were more likely to be tuned to HV AM rates ( < 40 Hz) when stimuli had pulses with HV rather than HC shapes.
6. Sharp temporal tuning was uncommon and found only in neurons with band-pass or low-pass characteristics.
7. Many neurons differed significantly in response to HV and HC stimulus sets. Maximum spike rate was more often elicited by an HV stimulus (74%) than by an HC stimulus (24%).
8. Differences in spike rates elicited by HV and HC stimuli were attributable to combinations of differences in the rise times and shapes of the pulses.

     

A study on the rate of water transport of the clam katelysia opima in relation to environmental conditions

1. The neutral red technique was employed to study the rate of filtration in Katelysia opima.
2. The weight specific water filtration was found to be greater for younger clams compared to the older ones.
3. The rate of water filtration increased with decreasing salinity.
4. Water filtration was found to increase as temperature increased, reaching a maximum at 35°C. but then sharply decreasing at 39°C.
5. Light had no significant effect on the rate of filtration.
6. Suspended matter was found to affect the rate of water filtration.
7. The rate of filtration was low at high pH and high in low pH.
8. The rate of water filtration was found to be faster during high tide than during low tide.
9. The presence of the parasitic crab, Pennotheris sp., in the mantle cavity of clams had a marked effect on the particle filtration.
10. Accidental cut of the siphon tips had no effect on the rate of filtration.

     

The correlation of the receptor potential with the light induced transient increase in intracellular calcium-concentration measured by absorption change of arsenazo III injected into Limulus ventral nerve photoreceptor cell

The light-induced membrane voltage response (receptor potential, ReP) and the absorption change of the intracellularly injected calcium indicator arsenazo III (arsenazo response) were recorded simultaneously in Limulus ventral nerve photoreceptor cells. A double pulse technique was applied for stimulation. After pressure injection of the indicator into the cell absorption changes were measured at 646 nm to obtain a measure of the changes of the intracellular calcium ion concentration.

1. The size of the arsenazo response increases with increasing intensity of the light stimulus. The intensity dependence of the size of the arsenazo response δA_max shows almost no correlate with the peak amplitude of the ReP, but correlates rather well with the time integral of the ReP.
2. Decreasing light adaptation (caused by prolongation of the repetition time of the pulse pairs) leads to an increase in size of the arsenazo response. Also here δA_max correlates better with the time integral of the ReP than with its peak amplitude.
3. Lowering the calcium concentration in the superfusate (from 10 mmol/l to ca. 40 Μmol/l) causes after ca. 10 min a drastical diminution of the arsenazo response to values below the noise level, and a less marked reduction in size of the ReP. The falling phase of the ReP is slower. After return to normal calcium concentration the arsenazo response recovers partly in ca. 50 min, while the ReP recovers faster.

The results show two opposite correlations between ReP and arsenazo response: Increase in size and duration of the ReP causes a greater transient increase of the intracellular calcium ion concentration. This in turn tends to reduce and shorten the ReP. Which effect dominates obviously depends on the conditions of the experiment: when the calcium concentration in the superfusate is reduced to ca. 40 Μmol/l a slow decrease of the ReP is accompanied by a small increase of the intracellular calcium ion concentration.     

Temporal discharge patterns of tectal and medullary neurons chronically recorded during snapping toward prey in toads Bufo bufo spinosus

In freely moving toads, the temporal discharge patterns of tectal and medullary neurons were observed during prey-catching.

1. Tectal T5.2 and T8.1 neurons displayed a premotor warming up firing that in the former was addressed specifically to prey orienting or snapping and in the latter generally to almost any kind of body movement.
2. The temporal discharge patterns of T5.2 neurons during snapping were different from those during orienting toward prey. Snapping started in the peak phase of warming up; firing was immediately terminated during the snap; thereafter some rebound activity was observed. Orienting started after the premotor warming up in the declining phase whilst the neuron kept on firing during orienting and then settled when the orienting movement was completed.
3. In toads which were not motivated to catch prey — comparabl to immobilized ones — the discharge frequency of T5.2 neurons toward a prey stimulus revealed no such warming up.
4. Because it is known that prey-selective T5.2 neurons are controlled by pretectal inhibitory influences, the following experiment was conducted: during recording a T5.2 neuron a pretectal lesion was applied ipsilaterally to the recording site. After a few seconds, the neuron showed a strong premotor wanning up in response to any kind of moving object, followed by prey-catching.
5. In the medulla oblongata, different H-type neurons of the hypoglossal nucleus displayed specific discharge patterns which resembled the tongue protractor and retractor muscle activities; a third type resembled the activity of the genio/sterno-hyoid muscle, which are suggested to stabilize the hyoid bone during snapping.
6. There were medullary M8-type neurons with properties similar to T8.1.
7. Snapping could be triggered by electrical stimulation of the optic tectum in the representation of the frontal visual field, but not by stimulation in the hypoglossal nucleus or the adjacent medial reticular formation.
8. A concept of a neuronal circuit for the coordination of tongue muscle contractions in response to prey is proposed.

     

Modulation of the ATP induced [Ca2+]c increase in as-30D hepatoma cells

• 1.1. The regulation of the increase in the cytosolic calcium concentration ([Ca²⁺]c) induced by extracellular ATP in AS-30D hepatoma cells was studied.
• 2.2. Homologous desensitization involving the refilling of intracellular calcium pools and the participation of protein kinase C was found.
• 3.3. Isoproterenol, forskolin and dibutyril-cyclic AMP also induced an increase in [Ca²⁺]c.
• 4.4. Interestingly, synergism was found for isoproterenol or forskolin and ATP.
• 5.5. The results suggest that there are two pathways for mobilizing [Ca²⁺] in AS-30D hepatoma cells; one is activated by ATP receptors and the other by cyclic AMP.

     

The anaerobic metabolism of malate of Saccharomyces bailii and the partial purification and characterization of malic enzyme

1. The main pathway of the anaerobic metabolism of l-malate in Saccharomyces bailii is catalyzed by a l-malic enzyme.
2. The enzyme was purified more than 300-fold. During the purification procedure fumarase and pyruvate decarboxylase were removed completely, and malate dehydrogenase and oxalacetate decarboxylase were removed to a very large extent.
3. Manganese ions are not required for the reaction of malic enzyme of Saccharomyces bailii, but the activity of the enzyme is increased by manganese.
4. The reaction of l-malic enzyme proceeds with the coenzymes NAD and (to a lesser extent) NADP.
5. The K _m-values of the malic enzyme of Saccharomyces bailii were 10 mM for l-malate and 0.1 mM for NAD.
6. A model based on the activity and substrate affinity of malic enzyme, the intracellular concentration of malate and phosphate, and its action on fumarase, is proposed to explain the complete anaerobic degradation of malate in Saccharomyces bailii as compared with the partial decomposition of malate in Saccharomyces cerevisiae.

     

Anémie du sujet âgé

Anaemia is common in patients older than 75 years. Patho-physiologic mechanisms of anaemia are similar to that involved in younger patients even though certain mechanisms are more common in the elderly. Practical decisional algorithms have to be used for the diagnosis of anaemia. Risk factors for anaemia have been identified in this age group: institutionalization, bad socio-economic conditions, co-morbid conditions. Particular emphasis is given to several points:

1. The clinical tolerance of anaemia is the most important feature to take into account in older patients, rather than the decline of the haemoglobin level. Even if the haemoglobin level is only slightly decreased, evaluation of anaemia should be performed by the geriatrician, including, if needed, the achievement of a bone-marrow aspiration.
2. Causes of anaemia are often multiple, due to frequent comorbid conditions and acute illnesses, associated with nutritional deficiencies, inflammation… Chronic kidney disease and dysthyroidia are also common.
3. Nutritional deficiencies (iron, folate and/or B12 deficiency) are frequent and their cause has to be identified.
4. The prevalence of myelodysplastic syndromes, which are clonal diseases, is probably underestimated in this age group.

     

An ecological study of the Ponggol Estuary in Singapore

1. An ecological study of the Ponggol Estuary was conducted from July 1965 to June 1966 and the seasonal data on physical, chemical and biological characteristics were presented.
2. The Ponggol River represents a short, narrow and shallow estuary in Singapore. The river mouth is open throughout the year and water from eastern Johore Straits drains in twice a day at high tide. The upper reach, however, is left exposed at low tide.
3. The Ponggol River was classified as a vertically and laterally homogeneous estuary and was found to exhibit a mesohaline to polyhaline environment.
4. Significant hydrological gradients from the river mouth 10 the upper reach were noted in the river system. Salinity, dissolved oxygen and pH increased towards the mouth of the river and other parameters such as nutrients, dissolved organic matter and turbidity increased towards the source.
5. Although the river received organic pollutants at the upper reach the estuary was able to discharge them fairly rapidly through regular flushing by the tides. The transient rise of organic matter did not appear to impart any serious affect on the biota in the estuary.
6. Over 98% of the phytoplankton consisted of diatoms, most of which were brought into the estuary from eastern Johore Straits. Freshwater forms were relatively few.
7. Phytoplankton biomass was considerably higher than the adjoining waters. and was reduced at the upper reach due to high turbidity of the water.
8. 80% of the zooplankton was composed of dinoflagellates,Difflugia, copepods and bivalve larvae dominating at all sections of the estuary.
9. Percentage composition of the zooplankton showed that dinoflagellates and copepod nauplii predominated at high tide whileDifflugia and bivalve larvae were abundant at low tide.
10. Zooplankton standing crop, in general, was higher towards the source at high tide but the reverse was found at low tide, i.e. standing crop increased towards the river mouth. This was attributed to the process of concentration.
11. Species composition of zooplankton was found to be more or less similar to that of the eastern Johore Straits.
12. The nekton consisted predominantly of small and juvenile fish. Close correlation of fish and copepods was found to be statistically valid and it was concluded that the fish entered the estuary to feed rather than to spawn.
13. The squids formed an important catch of the beach seine unit and were caught throughout the year.
14. The fish population could be grouped into four categories: estuarine components, euryhaline components, marine components and migratory components.
15. Benthic invertebrates were abundant. Commercially important species consisted of prawns,Metapenaeus andPenaeus, and crabs,Neptunus pelagicus andScylla serrata.
16. The river bed was inhabited predominantly by molluscs and the distribution resembled that of the sheltered shore of muddy-sand type.

     

The role of mitochondria in modifying the cellular inoc environment: Studies of the kinetic accumulation of calcium by rat liver mitochondria

1. The affinity of ATP-supported Ca²⁺ accumulation for both Ca²⁺ and ATP was determined from initial rate studies employing isolated rat liver mitochondria. TheK _m values for “free” Ca²⁺ and ATP were calculated to be of the order of 2 μM and 100 μM, respectively. TheK _m for ATP decreased as the Ca²⁺ concentration was increased.
2. The curve relating initial rates of Ca²⁺ accumulation to Ca²⁺ concentration was singmoidal in shape; values obtained for the Hill coefficient were in the range 1.5–1.9.
3. Concomitant with the ATP-stimulated accumulation of Ca²⁺, ATP translocation was itselt increased in the presence of Ca²⁺. This stimulation took place independently of Ca²⁺ accumulation.
4. Decreasing the pH of the incubation medium decreased the rate of Ca²⁺ accumulation. This inhibition was competitive in that the affinity of mitochondrial for Ca²⁺ could be altered. The maximal rate of accumulation did not change with change in pH.
5. The permeant anions inorganic phosphate and acetate stimulated the accumulation of Ca²⁺ in a non-competitive manner. Both theV _max and K_m varied when either of the anions were present.
6. The data are discussed in relation to the role that mitochondria play in controlling the cellular ionic environment.

     

Intracellular amino acids and protein synthesis in Hela cells

1. When the intracellular amino acid pool is prelabelled and subsequently chased in non-radioactive medium, the radioactivity of the amino acid pool is not found to have been incorporated into protein.
2. Leucine transport into Hela cells is reduced in the presence of 10 mM valine in the medium. This results in a lower specific radioactivity of leucine in the intracellular amino acid pool. However, neither the overall rate of protein synthesis nor the incorporation of radioactive leucine into protein is affected.

From these experiments it is concluded that incoming amino acids entering the intracellular amino acid pool are not used for de novo synthesis of protein.     

Effects of membrane-perturbing drugs and noradrenalin on cAMP accumulation and red cell water content in carp (Cyprinus carpio) red cells

• 1.1. Carp red cells were treated with drugs that affect the cell membranes. The water content of the cells and the accumulation of cAMP in the cells were measured in normoxia and in hypoxia using non-stimulated and adrenergically stimulated cells.
• 2.2. WGA, DIDS + CCCP and A23187 increased the water content of nonstimulated normoxic cells.
• 3.3. In hypoxia ouabain and DIDS + CCCP increased the water content but cytochalasin B, NPM, DIDS, CCCP and A23187 + CA²⁺ abolished the hypoxia-induced swelling.
• 4.4. Any membrane perturbation induced some cAMP formation, Sophora and Anquilla lectins being most potent.
• 5.5. Also in adrenergically stimulated cells, membrane perturbation generally increased cAMP formation.
• 6.6. However, cAMP accumulation diminished in cells treated with cytochalasin B, CCCP and DIDS + CCCP.
• 7.7. The adrenergic swelling of carp red cells was reduced in normoxia by DIDS. NPM and CCCP increased the adrenergic swelling in normoxia to hypoxic level.
• 8.8. In hypoxia WGA and Anquilla lectin decreased the swelling.

     

Immunocytochemical studies of the Gi Protein mediated muscarinic receptor-adenylyl cyclase system

The localization of three key signal transduction components was indicated in rat heart tissue by immunocytochemical and histochemical experiment. It was shown that:

1. The M2 muscarinic receptors are localized along outer cell membranes and T-tubule membranes of cardiomyocytes but additionally at membranes of endothelial cells and fibroblasts.
2. G_iα was found along outer cell membranes of cardiomyocytes and other cells of the heart and also inside the cells of the perinuclear space in close contact to the nuclei envelope and the endoplasmic reticulum membranes. G_oα were found to be associated mainly in atrial tissue, especially at the nerval (neuronal) endings located among the cardiac muscle cells. This was shown in parallel incubation with specific neuronal antibody as a marker for these structures.
3. Adenylyl cyclase was localized along the sarcolemma and the T-tubule membranes in normal cardiomyocytes of rat and guinea pig hearts. Under ischemic conditions, the adenylyl cyclase was also seen in junctional sarcoplasmic reticulum membranes. The reasons for this changed localization need further elucidation. Binding of the adenylyl cyclase within the molecular structure of the membrane or variation of the marker penetration remain to be clarified.