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.

     

Osmoregulation in immature atlantic salmon (Salmo salar L.) following transfer from sea-water to fresh water

• 1.1. Immature Atlantic salmon post-smolts weighting approximately 150 g were transferred abruptly to fresh water (FW) after 5 months in sea water (SW).
• 2.2. Losses of ions and gain of body water are reversed after 3 days with about 10–12 days taken for complete FW adaptation.
• 3.3. Immediately on transfer from SW to FW, immature salmon take up sodium at 45 μmol/kg/hr, about one-third the rate observed in maturing salmon on their spawning migration.
• 4.4. The sodium uptake rate increases to that of maturing salmon after 2 days in freshwater. Differences in the osmoregulatory ability of immature and maturing salmon are discussed.

     

Lipid reserves and body composition in postreproductive anurans

• 1.1. The mass of lipid extracted from 13 species of adult anurans collected immediately after reproduction differed between sexes.
• 2.2. Covariance analyses of lipid mass with body mass as the covariate indicated that females had significantly more lipid than males; the proportion of lipid as a percentage of body mass was correlated significantly with water content of males, but was not correlated significantly in females.
• 3.3. Heat of combustion of extracted lipid increased among species in relation to the timing of reproduction; species that bred later in the year had a greater energy content per gram of lipid.
• 4.4. Major changes in composition during growth and metamorphosis of bullfrogs (Rana catesbeiana), include increases in lipid and energy content per gram of tissue interrupted by a severe decline of lipid reserve during metamorphosis.

     

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.

     

Diurnal variation in Sagar Lake,Sagar (India) I studies in the deep water area

1. A diurnal study of inland fresh water lake has been made with respect to physical and chemical properties and the plankton.
2. Chlorides have followed the total carbonates while dissolved oxygen and pH have shown no relation.
3. Microcystis has followed no definite pattern of diurnal movement.
4. All crustaceans, some of the rotifers andTrachelomonas perform considerable diurnal movement in the course of a twenty four hour period.

     

Field studies on the variability of populations of Aster tripolium L. in relation to saltmarsh zonation

1. Aster tripolium is a very variable species of which a number of types have been described both on a morphological and ecological level.
2. In permanent plots along the height gradient in the salt marsh it appeared that differences in the A. tripolium subpopulations occur.
3. In the lowest zone of the marsh, dominated by Spartina anglica (Spartinion), the mean life expectancy of individual adult plants is relatively high but it is relatively low for seedlings.
4. In the higher zones of the marsh the mean life expectancy is relatively lower, but for the seedlings it is higher (Puccinellion maritimae).
5. In the Spartina-zone individual A. tripolium plants have about twice the number of shoots as the plants from the other zones.
6. The plants from the Spartina zone produce per individual more generative shoots (absolute) but these have less capitula than elsewhere in the marsh. The number of ripe seeds per head is almost constant everywhere in the marsh.

     

Improvements of the membrane filter method for DNA:rRNA hybridization

We describe and recommend the following improvements of DNA:rRNA membrane filter hybridization methods. One of our aims was to avoid DNA release from filter discs during hybridization.

1. Our hybridization conditions are 2 SSC in aq. dest., with 20% formamide, 50 C, overnight for 16 hr.
2. Duplexing is over in 8–10 hr.
3. Formamide has to be very pure (O.D.≤0.2/cm light path at 270 nm).
4. RNAase treatment: 250 μg/5 ml 2 SSC/filter at 37 C for 1 hr.
5. Our conditions for stepwise thermal denaturation are: 5°C steps from 50C to 90C in 1.5 SSC in 20% formamide.
6. Single-stranded DNA, fixed on membrane filters, and stored in vacuo at 4C, can be used reliably for hybridization for up to 20 months.
7. Concentrated DNA in 0.1 SSC, quick-frozen at ?50 C and stored at ?90 C for up to 2 years can be used for hybridization without much change.
8. A CsCl gradient purification step yields much purer DNA, but increases the release of DNA from filters by about 20%. Filters with 20% more DNA is a compensation.
9. rRNA can be stored for 20 months in SSC or 2 SSC at ?12C without changing the hybridization results.

     

Uptake of guanine by synchronizedChlorella fusca

1. The transport of guanine in autospores of light-dark synchronizedChlorella fusca was studied using radioactive guanine in the concentration range of 4 nM to 50 μM.
2. The transport system was constitutive, it had high specificity for the permeant, and theQ ₁₀ value was in the range of 1.5 to 2.2. At concentrations lower than 0.2 μM the half saturating constant, S_0.5 was 1 μM both for cells kept in dark and cells kept in light. At higher concentrations the S_0.5 of darkened cells was about 0.23 μM, while that of illuminated cells was unchanged. Only above 0.2 μM guanine did illumination of the cells or addition of glucose increase the transport rate.
3. Guanine which had accumulated did not leak out at temperatures below 45°C or by treatment with 10 μM dinitrophenol, which completely inhibited transport. Furthermore, the accumulated guanine did not exchange with exogenous guanine.
4. The guanine accumulated, more than 10⁵-fold over the external concentration, showing that the transport, was active.
5. The initial transport rate per cell revealed annual fluctuations.

     

Purification and some particular kinetic properties of rat spleen cytosolic deoxynucleotidase

• 1.1. Rat spleen cytosolic deoxynucleotidase was purified 40,000-fold to almost homogeneity and had a specific activity of 3000 μmol/min per mg.
• 2.2. Molecular mass of the native enzyme was 45 kDa. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that the native enzyme comprises two identical 27-kDa subunits.
• 3.3. Specific enzyme activity increases with increasing concentration of enzyme protein and approaches a plateau at high enzyme concentrations.
• 4.4. Enzyme activity increases gradually and nonlinearly with increasing concentration of enzyme in the low concentration range. Above a certain concentration the increase attains a maximal and constant slope.
• 5.5. The kinetic properties can be explained by assuming dissociation of the enzyme into subunits with low or no activity.

     

Sodium balance in adult atlantic salmon (Salmo salar L.) during migration into neutral and acid fresh water

• 1.1. In sea-water, adult salmon (S. salar) exchange an average of 12.6% of total body sodium/hr.
• 2.2. Following transfer to fresh water sodium uptake follows Michaelis-Menton kinetics. F_max = 2.40 mmol Na/1 ECF/hr, K_m = 0.26 mmol Na/1. The uptake system is fully activated immediately following transfer to fresh water.
• 3.3. Post smolts adapted to sea-water for 3 months take up sodium at only one third of the rate of adult fish following return to fresh water.
• 4.4. The concentration of prolactin in the plasma is low in sea-water adapted fish and does not rise during the first 8 hr in fresh water.
• 5.5. At pH 5 sodium uptake is reduced by almost 90%, even in the absence of aluminium, but recovers immediately on return to neutral water.
• 6.6. At pH 5 and 20 μmol Al/1 there is little further effect on sodium uptake but after 6 hr in aluminium the inhibition of sodium uptake continues after return to neutral aluminium fresh water and uptake is only 50% of normal 24 hr later.

     

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.

     

Ribosome inactivating protein-like activity in seeds of diverse Cucurbitaceae plants

• 1.1. Seed extracts of 20 plants species belonging to the family Cucurbitaceae were examined for their ability to inhibit protein synthesis in rabbit reticulocyte lysate and induce mid-term abortion in mice.
• 2.2. Eleven extracts were found to inhibit protein synthesis by about or over 90%, seven extracts produced about 80% inhibition, one caused about 70% inhibition and one brought about approx. 40% inhibition, when the extracts were tested at a final concentration of 10 μg per ml.
• 3.3. All of the seed extracts possessed potent mid-term abortifacient activity.
• 4.4. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the seed extracts disclosed the existence of a Coomassie Blue-stainable band with a mol. wt of ca 30,000 Da. This band probably accounts for the protein synthesis inhibiting and mid-term abortifacient activities.
• 5.5. There was a similarity in the electrophoretograms of seed extracts of plants belonging to the same genus.

     

Neurotransmitters of cephalopods

1. ACh, dopamine, noradrenaline, 5-HT,l-glutamate, and GABA are widely distributed in cephalopods and probably all function as neurotransmitters; octopamine also occurs and at one site is known to act as a neuromodulator.
2. Several peptides are also present, as well as nitric oxide synthase.
3. In the brain and sense organs cholinergic, aminergic, serotonergic and glutamatergic systems seem to be the most important.
4. ACh is also active in the gut, vascular system and some body muscles: it is generally inhibitory. The ACh receptors are similar to the vertebrate nicotinic type.
5. The catecholamines are important in the gut and vascular system: they are generally excitatory. The NA receptors are like the α-adrenergic subtype of vertebrates, but the nature of the DA and OA receptors is less certain.
6. 5-HT is important in the gut but is endogenous in some chromatophore nerves and acts on receptors that seem like the vertebrate 5-HT₁ type.
7. l-glutamate is an excitatory transmitter at the chromatophore (and probably at other) nerve-muscle junctions and is an extremely strong candidate for being the excitatory transmitter at the squid giant synapse. There are NMDA receptors on Schwann-cells but the receptors on neurons and muscles are like the vertebrate kainate type.
8. Little is known about the mode of action of cephalopod peptides; nor has it ever been shown that they co-exist with conventional transmitters in these animals.
9. The structure of one (FMRFamide) receptor has been elucidated, but apart from this nothing is known of the molecular biology of receptors in cephalopods.

     

Correlation of the effects of citric acid cycle metabolites on succinate oxidation by rat liver mitochondria and submitochondrial particles

1. Succinate dehydrogenase is inhibited by citrate and β-hydroxybutyrate in a complex manner, both in mitochondria and submitochondrial particles. Kinetics of inhibition in the particles points to a competitive component in the mechanism involved.
2. Pyruvate, α-ketoglutarate, malate, and glutamate stimulate oxidation of succinate by mitochondria.
3. Stimulation by α-ketoglutarate and glutamate is not influenced by the presence of rotenone.
4. Stimulation by pyruvate is higher in the absence of rotenone and increases significantly in the presence of K⁺ and valinomycin. Pyruvate supplies in mitochondria reducing equivalents for malate dehydrogenase operating in the reverse direction-reduction of oxaloacetate to malate.
5. Stimulation by malate is higher in the presence of rotenone.

     

Acid-base titration across the plasma membrane ofMicrococcus denitrificans: Factors affecting the effective proton conductance and the respiratory rate

The object of this work was to measure the effective proton conductance of the plasma membrane ofMicrococcus denitrificans under various conditions and to investigate possible connections between respiration and proton translocation.

1. Pulsed acid-base titrations of suspensions ofM. denitrificans in a medium containing the permeant thiocyanate ion, or when K⁺ ion permeability was induced by valinomycin in a KCl medium, showed that the normal effective proton conductance of the membrane system was less than 1 μmho/cm².
2. A pH-overshoot artefact was suppressed by adding carbonic anhydrase.
3. The effective proton conductance was increased by the uncoupler FCCP in the same concentration range as was required to stimulate respiration. Concentrations of FCCP above 1·5 μM inhibited respiration after an initial stimulation.
4. The effective proton conductance in presence of 2 μM FCCP was at least 17 μmho/cm².
5. The quantitative relationships between the respiratory rate, the stoichiometry of respiration-driven proton translocation, and the effective proton conductance of the membrane of the cells are compatible with the suggestion that stimulation of respiration by FCCP is due to a release of back-pressure exerted by a protonmotive potential on the respiratory chain system in the membrane. Only one amongst other possible explanations of the stimulation of respiration by FCCP is, however, excluded.

     

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.

     

Resource allocation by plants under air pollution stress: Implications for plant-pest-pathogen interactions

Plant growth depends on the coordinated acquisition and allocation of carbon, water, and nutrient resources to the major plant organs (root, stem, leaf, flower, and fruit) and to the major classes of metabolic function (vegetative growth, maintenance, defense, and reproduction). Air pollutants like SO₂, NO₂, and O₃ can directly damage plant tissues and disrupt normal patterns of resource acquisition and allocation. These disruptions in turn potentially will influence the plant’s ability to defend itself against pests and pathogens. This review summarizes the quantitative and qualitative changes that have been observed when plants are exposed to low levels of SO₂, NO₂, and O₃; the following generalizations emerge:

1. Root biomass is reduced more than shoot biomass in plants exposed to SO₂ or O₃, but NO₂ does not appear to induce the differential suppression of above-versus below-ground organs.
2. Quantitative allocation to leaves increases and to stem decreases under SO₂ pollution regimes; too few data are available to generalize about O₃ or NO₂ effects on leaf: stem ratio.
3. Root carbohydrate concentrations sometimes increase and sometimes decrease after SO₂ or O₃ fumigations. Leaf nitrogen concentrations tend to decrease after exposure to air pollutants, and leaf carbohydrate concentrations can increase or decrease. Too few data on leaf concentrations of lipids and secondary chemicals are available to justify any generalizations on pollutant responses.
4. Reproduction is suppressed by O₃, SO₂, and NO₂, with O₃ appearing to have the most marked effects. Seed lipid and protein composition can be altered by exposure to pollutants. While both quantitative and qualitative changes in plant resource allocation after exposure to pollutants are common, the importance of these diverse changes for plant-pest and plant-pathogen interaction requires more comprehensive study. Ideally, the time course of plant growth and of metabolite pools critical to particular pests or pathogens should be followed in plants exposed to realistic pollutant regimes and related to pest or pathogen performance on the treated plants.

     

Uptake of radionuclides by some aquatic macrophytes of Ismailia Canal,Egypt

1. Uptake of the radionuclides Co-60, Cs-134, Sr-89 and P-32 by different aquatic plants present in Ismailia Canal was found to increase at increasing initial concentrations of the radionuclides in the water.
2. Maximum uptake was reached by most of the aquatic plants after different periods of contamination, ranging from 1 to 4 days.
3. Ceratophyllum was found to be the most favourable biological indicator for strontium radioisotopes in concentrations ranging from 0.5 to 10 µCi/1 and for contamination periods up to 16 days.
4. Elodea was found to serve as a biological indicator for either strontium or phosphorus radioisotopes for limited contamination periods of 2 to 16 days for Sr and to 2 days for P isotopes.
5. Potamogeton was also found to serve as a biological indicator for limited contamination periods for strontium, phosphorus and cesium radioisotopes: 0 to 2 days for Sr, 2 to 16 days for P. and 0 to 2 days for Cs isotopes.

     

A novel method for regenerating plants from mesophyll protoplasts of Arabidopsis thaliana line WS

A procedure has been developed for the successful regeneration of plants from mesophyll protoplasts of Arabidopsis thaliana line WS. The protocol is an improved version of that of Damm and Willmitzer (1988). The main changes in original procedure are as follows:

1. A mixture of Cellulase Y-C (0.5%) and Pectolyase Y-23 (0.05%) is used for the isolation of protoplasts. Use of these enzymes reduces the incubation time to 50 min.
2. α-Naphthaleneacetic acid is used as the auxin throughout cultures of protoplasts and calli.
3. Protoplasts and calli are incubated under dim white light (0.8–8 μW/cm²) during culture.

With these modifications, we were able consistently to obtain regenerated shoots from about 70% of calli that had been transferred to shoot-forming medium even though the plating efficiency was rather low (about 0.5–1.5%).     

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.