Studies on intestinal protease: Isolation,purification and effect of dietary proteins on alkaline protease activity of the air-breathing fish,Clarias batrachus (Linn.)

1. The effect of dietary protein levels on the proteolytic activity in the intestines of the air-breathing fish, Clarias batrachus (Linn.) has been studied
2. Activity of proteolytic enzymes increased significantly in fishes maintained with a 50% protein diet from those maintained with a 25% protein diet; still higher dietary protein percentage showed no further stimulation of enzyme activity.
3. In a study on the determination of sub-cellular localisation, it has been found that protease activity is more prominent in lysosomes than in other organelles of the cell.
4. A sixty fold purification of alkaline protease from the intestine of Clarias batrachus has been achieved by ion exchange chromatography on DEAE cellulose which has been further checked by polyacrylamide gel electrophoresis.

     

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.

     

The use of toxicokinetics for the safety assessment of drugs acting in the brain

Pharmacological and toxicological studies undertaken on drugs that affect the brain are frequently performed in disparate species under various experimental conditions, at doses often greatly in excess of those expected to be administered to humans, and the findings are extrapolated implicitly or explicitly with scant regard to differences in the biodisposition of the drugs. Such considerations are necessary since:

1. Species;
2. Strain;
3. Gender;
4. Route;
5. Dose;
6. Frequency and time of administration;
7. Temperature;
8. Coadministration of drugs; and
9. Surgical manipulation

are but some of the factors that have been shown to influence the kinetics and metabolism of drugs. This article, using MDMA and other phenylethylamines as examples, provides evidence for the need to measure the exoosure of the drugs and their active metabolites in blood and brain (toxicokinetics) in order that conclusions based only on dynamic, biochemical, or histological evidence are more pertinent. Further, the combined use of toxicokinetic-dynamic modeling can lead to a better appreciation of the mechanisms involved and a more useful approach to the calculation of safety margins.     

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.

     

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.

     

The role of touch,pressure and nociceptive mechanoreceptors of the leech in unrestrained behaviour

1. The maximum force exerted against an isometric force transducer by 6 leeches weighing 2.6–3.7 g, as they squeezed through apertures of different widths varied inversely with aperture width.
2. T cells in the leech skin code for velocity of indentation, not pressure or displacement. The frequency with which T cells fire is best described by two log functions, one for low, another for fast indentations. T cells responded to indentation velocities down to 10 μms^?1.
3. The average threshold pressure for 5 P cells was 150 kPa and for 5 N cells was 521 kPa.
4. We conclude from these data that when leeches explore their mechanical environment and initiate contact with external objects, the threshold pressure for N cells is rarely crossed. Of the three classes of mechanoreceptor, T cells are the main modality through which leeches obtain contact information, though P cells may occasionally be recruited for local pressure peaks.

     

Antioxidant enzymes and trace elements in hemodialyzed patients

Antioxidant enzymes together with trace elements in 26 patients with chronic renal failure treated with hemodialysis and 25 healthy subjects were investigated. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSHPx) in plasma and erythrocytes were examined immediately before and after hemodialysis. The results are summarized as follows:

1. A significant decrease in plasma SOD, CAT, and GSHPx and erythrocyte GSHPx were found in patients before hemodialysis.
2. Erythrocyte CAT and GSHPx were significantly lower in the patients after hemodialysis than in the controls.
3. Plasma GSHPx was significantly higher after a single hemodialysis than before hemodialysis.
4. A good correlation between erythrocyte SOD and copper (Cu) in patients before hemodialysis was found.
5. A good correlation of GSHPx in erythrocytes and plasma was found before hemodialysis, whereas an even better correlation was found after hemodialysis.
6. Abnormalities of trace elements were found in hamodialyzed patients.
7. There is indirect evidence for increased oxidizing stress in uremic patients with hemodialysis. Dialysis treatment may improve some abnormalities (e.g., Hb, P), but may also induce some deleterious effects of free radicals or lipid peroxidation.

     

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.

     

Effects of Environmental Perturbations on Short-Term Phytoplankton Production off Lawson's Bay,A Tropical Coastal Embayment

1. The phytoplankton cycle off Lawson's Bay, Waltair follows a bimodal pattern with a major peak during March–May; a minor peak during October–November months and with a low production during the summer months i.e., June–August.
2. During the summer months of 1957, 1958, 1960 and 1962 dumping of dredged spoil from the entrance channel of the harbour into the sea resulted in a natural enrichment of waters.
3. Following this enrichment, there was a qualitative and quantitative increase in the phytoplankters thus leading to the development of a bloom.
4. Only Thalassiosira subtilis and Chaetoceros curvisetus commonly bloomed during the four years.
5. The increase in gross production which varied from 3–13 fold and the high photosynthesis-respiration ratios 5.1 to 10.5 indicated that the bloom populations were in a healthy state.
6. The decrease of the populations to the initial levels suggests that some unknown factor, other than those investigated must have been operating.
7. Consequences of eutrophication of different origins on stimulation of phytoplankton production are briefly discussed.

     

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.

     

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.

     

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.

     

EPR characterization of oxide supported transition metal ions: Relevance to catalysis

EPR spectroscopy is a powerful tool to identify at a molecular level, the different steps of catalyst preparation, and of catalytic reactions:

1. Deposition of paramagnetic transition metal ions onto a support is monitored, and the coordination sphere of the metallic center is characterized by EPR.
2. The catalyst is also characterized after activation (thermal oxidation or reduction):

- the distribution among the different sites in zeolites can be determined;

- the dispersion of the active phase may be appreciated;

- the unsaturation degree of the active site may be evaluated using probe molecules such as water or¹³C enriched carbon monoxide.

1. The catalytic mechanisms can be investigated by studying the elementary steps of the catalytic reaction, as illustrated for methanol oxidation over Mo/SiO₂ catalysts whose EPR results have extended the reaction mechanism proposed on the basis of kinetic data. In addition, reaction intermediates may be isolated inquasi-in situ conditions as in the case of olefin oligomerization catalyzed by Ni/SiO₂ systems.

     

A comparison of serum retinol concentration between human and different species of normo and hypercholesterolemic nonhuman primates fed semi-purified diets with defined amounts of vitamin A

• 1.1. Serum retinol and total cholesterol concentrations were determined in several species of nonhuman primates fed semipurified diets. Two species of Old World and three species of New World nonhuman primates were examined.
• 2.2. Retinol levels were significantly lower (up to four-fold) in the serum of the smaller New World than the larger Old World animals and the difference could not be explained by differences in dietary make-up.
• 3.3. Cholesterol levels were not different between the groups but differed within a species when type of dietary fat was altered.
• 4.4. Differences in circulating levels of retinol may reflect differences in levels of retinol binding protein between the groups.

     

Time-course of changes in plasma levels of trace elements after thrombolysis during the acute phase of myocardial infarction in humans

It has been suggested that the injury induced by reperfusion of the ischemic myocardium could result, in part, from the cytotoxic effects of oxygen free radicals. Since various trace elements are involved in several of the reactions leading to free radical production, we have measured plasma levels of copper, zinc, selenium, and iron:

1. In 18 patients (mean age 60 yr old) subjected to thrombolytic therapy within 6 h after the onset of a myocardial infarction (G1);
2. In 16 patients with coronary artery disease, but without a history of a previous myocardial infarction (MI) (mean age 50 yr old, G2); and
3. In 50 healthy volunteers divided into two subgroups according to age (mean age 33 yr old, G3 and 55 yr old, G4).

Plasma myosin levels were used to estimate quantitatively the extent of the infarcted mass. Plasma trace element levels were measured in blood samples following centrifugation and storage at ?80°C. The main results were as followed: In G1 patients who have been subjected to thrombolysis, an important release of myosin was measured in plasma, with a peak at D6 (1678 vs 95 μU/L at H0). In those G1 patients after MI:

1. A significant increase in plasma copper levels was observed from day 4 to day 10 postinfarction (×1.15 in reference to the baseline data at H0);
2. A decrease in plasma zinc levels was observed and was maximum 12 h after the onset of the thrombolytic treatment;
3. A decrease in selenium concentration was observed in G1, as well as in G2 patients, compared to the control groups (80% of G3 and G4 values); and
4. A significant decrease in plasma iron levels was observed in G1 (67.8% of G3 and G4 control values) and was significant from H0 to day 7 (p<0.01).

In conclusion, this study underlined the time-course evolution of plasma trace element levels in the followup of patients who have been subjected to thrombolysis following a MI and the potential prognostic implication of such variations.     

Hair zinc and copper concentrations and zinc: Copper ratios in pediatric malignancies and healthy children from Southeastern Turkey

Twenty-eight healthy and forty-six children with malignancy admitted to the Pediatrics Oncology Clinic and Outpatient Clinic of Dicle University, Medical Faculty, Department of Pediatrics in 1989–1990 were included into this study. The children under investigation were divided into three groups. Group 1 consisted of 28 healthy children. Twenty-eight children with newly diagnosed malignant disease, before the start of treatment, formed Group 2. Group 3 consisted of eight of these 28 children together with 18 other children with malignancy in complete remission.

1. Mean hair zinc and copper levels, and zinc/copper values for Group 2 were found to be 100.7±6.8 μg/g, 11.0±0.8 μg/g, and 10.0±0.8, respectively.
2. The corresponding values for Group 3 were obtained as 143.5±6.3 μg/g, 7.6±0.4 μg/g, and 20.2±1.4.
3. The values for Group 1 were 142.6±4.5 μg/g, 8.4±0.5 μg/g, and 19.0±1.4, respectively.
4. The differences between control-active-phase and active-phase-remission groups were statistically significant (p<0.01), whereas almost the same values were obtained for the control and remission groups (p>0.05). These results were valid in cases where the comparisons of these three groups were performed also for the subgroups.

It was shown that hair zinc and copper levels and zinc/copper values are useful parameters for the diagnosis and treatment of the children with malignancy because of the fact that hair reflects the integrated trace element concentrations of the body, not the present one.     

Survival,osmoregulatory ability,and respiration of idotea chelipes (Crustacea,Isopoda) from lake veere in different salinities and temperatures

1. An ecological and physiological study ofI. chelipes from Lake Veere, The Netherlands, was made.
2. Both osmoregulatory capacity and survival decrease with increasing temperature as well as with decreasing salinity.
3. Respiration experiments suggest that the need of energy by osmoregulatory activity may be supplied at the cost of other physiological processes, at any rate at temperatures of 10°C and higher.
4. It may be expected that, if temperatures higher than 15°C and salinities lower than 8‰ coincide, the population ofI. chelipes will be affected negatively.

     

Total recovery of O2 evolution and nanosecond reduction kinetics of chlorophyll-a II + (P-680+) after inhibition of water cleavage with acetate

Oxygen evolution and reduction kinetics of the photooxidized Chl-a_II ⁺ have been measured in oxygen-evolving complexes from the thermophilic cyanobacterium Synechococcus sp.

1. Incubation of PS II particles with acetate resulted in an inhibition of oxygen evolution and a retardation of the Chl-a_II ⁺=reduction kinetics from the nanosecond range to the microsecond range, indicating a modification of the donor side of photosystem II (PS II).
2. After the first two flashes given to a dark-adapted, acetate treated sample, Chl-a_II ⁺ was re-reduced with a half-life time of 160 μs by a component of the donor side of PS II. Under repetitive excitation Chl-a_II ⁺ was re-reduced in 500 μs by electron back reaction from the primary acceptor Q_A ^- (X-320^-). Obviously, in the presence of acetate only two electrons are available from the donor side.
3. Both oxygen evolution and nanosecond reduction kinetics of Chl-a_II ⁺ were restored to the control level when acetate was removed.
4. The results indicate a tight coupling between O₂ evolution and nanosecond reduction kinetics of Chl-a_II ⁺.
5. The reversible inhibition is probably due to a replacement of Cl^- by acetate within the water splitting enzyme.
6. Due to its strongly retarded kinetics, the reversibly modified system may facilitate investigations of the mechanism of the donor side.

     

Effet des androgènes sur l’activité sportive

Androgenic hormones seem to be of beneficial effects on sports performance:

- they increase motivation, will, aggressiveness, resistance to the stress and to the fatigue, leading to an increase of the training quantity,

- they increase bone mineralization and probably mechanical resistance,

- they stimulate the bone marrow and so, with the erythropoietin, the erythropoiesis,

- they increase the tendancy to hyperglycemia, but with a decrease of the tolerance to the glucose,

- they stimulate the fatty acids mobilization from the adipose tissue, for their utilization in the muscle during the exercise,

- they participate, for the trained sportmen, to a better gestion of the muscle glycogen storage: their utilization during exercise is decreased,

- they increase the lean body mass, with an increase of the protein synthesis and a decrease of the protein catabolism, leading also to an increase of the muscle force under training. There is no beneficial effect upon the tendons,

- they have an immunomodulation action.

     

Experimental problems using electronic particle counters

The following scheme lists the problems which have been and will continue to be encountered in grazing experiments on natural particle assemblages. In some cases there are solutions, but many of the problems listed below remain intractable. To solve them will require innovative approaches, most probably combining the use of particle counters with other techniques to approach the problem to be solved from several angles at once. Variance between samples due to

- statistical causes (especially caused by large particles) and

- differences between experimental bottles and between experimentals and controls at start of experiment. Shifts in particle distribution during experiment due to

- increase in small particles

- growth of bacteria and other organisms due to excretion of grazers and

- breakage during handling by grazers into fragments. Processes, independent of grazing, leading to changes in particle size distribution due to

- primary production

- grazing by additional components and

- bacterial growth on detrital particles and the formation of detrital flocs.