Human spleen cathepsin D: Its characterization and localization in human spleen

• 1.1. The cathepsin D was purified 1830-fold under mild conditions by a rapid procedure, based on two-step affinity chromatography.
• 2.2. Its molecular weight, amino acid composition and substrate specificity were shown to display minor differences from materials of other origins.
• 3.3. Inhibition with thiol compounds was found to be a specific phenomenon of the cathepsin D from the human spleen.
• 4.4. Production of antiserum specific for purified cathepsin D was demonstrated by immunodiffusion test, an immunoadsorbent column and immunoblotting of the crude enzyme in SDS gel.
• 5.5. In an immunocytochemical study, the antigenic sites for this enzyme were found to be localized in the reticuloendothelial system of the human spleen.
• 6.6. The role of this enzyme in human spleen cell was discussed.

     

Phosphate metabolism during muscular contraction in starved frogs (Rana catesbeiana)

• 1.1. The muscle tension and the state of high-energy phosphate metabolism during contraction of the sartorius muscle in frogs (Rana catesbeiana) starved for 1–5 months was studied by in vivo³¹P-NMR spectrometry.
• 2.2. Muscle tension began to decrease after 2-month starvation compared with the control group and decreased to about one-third of the control value after a 5-month starvation.
• 3.3. Muscle contraction induced by electrical stimulation or the use of anaerobic perfusion fluid did not decrease the concentration of creatine phosphate (PCr) or β-ATP, and only negligibly changed the PCr/Pi ratio from starvation.
• 4.4. These results suggest a decrease in creatine kinase activity in the muscle of starved frogs.

     

An enzyme-inhibitor complex of cathepsin L in the white muscle of chum salmon (Oncorhynchus keta) in spawning migration

• 1.1. A cathepsin L-inhibitor complex was purified from the white muscle of chum salmon (Oncorhynchus keta) by a series of chromatographies on DEAE-Sephadex, con A-Sepharose and Sephadex G-150.
• 2.2. The mol. wt of the complex was estimated to be 50,000 by gel filtration. The complex per se showed little activity of cathepsin L, but it became active when incubated at an acidic pH.
• 3.3. SDS-PAGE analysis and an experiment of activation by acidification indicated that the complex consisted of the 37 or 30 kDA-form of cathepsin L and the 15 kDa-endogenous cysteine protease inhibitor.
• 4.4. The enzyme-inhibitor complex was considered to be formed when cathepsin L leaks out of the lysosome in vivo or is freed from the lysosome when the tissue is artificially destroyed.

     

Changes in composition and proteolytic enzyme activities of artemia during early development

• 1.1. The proximate composition, total and free amino acids, and proteases of Artemia nauplii were determined during early development.
• 2.2. Moisture increased from 71.0% to 80.8%, crude protein decreased from 13.2% to 8.8%, crude fat and ash varied slightly.
• 3.3. The total amino acids decreased. Free amino acids changed in three patterns.
• 4.4. Trypsin, chymotrypsin, carboxypeptidase A, B and cathepsin B and C increased in activity. The activity of trypsin was lower, while cathepsin B and C were the highest.
• 5.5. The protease activities were maximal at pH 7.5 and 8.0, and at 45°C on casein.
• 6.6. The optimal pH for carboxypeptidase A was 4.0, for carboxypeptidase B was 4.5, for trypsin and chymotrypsin were 7.0–7.5. The protease(s) active at pH 9.0–9.5 were to be determined.

     

Varying amounts of stretch stimulus regulate stretch-induced muscle hypertrophy in the chicken

• 1.1. The effects of different amounts of passive stretch per day and number of days of stretch on muscle hypertrophy in the chicken patagialis (PAT) muscle were determined.
• 2.2. Stretch for 24 hr per day (h/d) resulted in a more rapid hypertrophy both on a wet and dry tissue basis (P < 0.001) than stretch for 4 h/d.
• 3.3. Stretch increased PAT weight 43% and 25% in 24 h/d and 4 h/d treatments, respectively, after 10 days of stretch, but by day 25 of stretch there was no difference between treatments.
• 4.4. In a second experiment, the PAT muscle was hypertrophied and then the effects of intermittent stretch (4 h/d) on regression of hypertrophy (muscle atrophy) were investigated.
• 5.5. Intermittent stretch (4 h/d) for 5 and 10 d significantly (P < 0.001) inhibited regression of hypertrophied muscle.
• 6.6. The results of the present study indicate that stretch-induced hypertrophy can be modulated by varying the amount of stretch applied per day.
• 7.7. Intermittent stretch can be used to inhibit the regression which occurs when a continuous stretch stimulus is removed.
• 8.8. Intermittent stretch is a useful model for investigating mechanisms of muscle hypertrophy and inhibition of muscle atrophy.

     

Activity of some enzymes involved in the metabolism of polyamines in the liver of streptozotocin-diabetic rats

• 1.1. The effect of diabetes on some enzymes of polyamine metabolism was studied in male rats 1–12 days after administration of streptozotocin.
• 2.2. Hepatic ornithine decarboxylase activity decreased in the first days after the administration, but increased thereafter. The decrease was not due to an alteration of the ODC-antizyme concentration, nor to a posttranslational modification catalyzed by transglutaminase.
• 3.3. S-adenosylmethionine decarboxylase and ornithine transaminase were both increased.
• 4.4. Spermicline acetyltransferase activity was practically unchanged, while its inactivating factor was markedly decreased.

     

Physical conditioning: Effect on the myoglobin concentration in skeletal and cardiac muscle of bar-headed geese

• 1.1. A 12 week program of treadmill exercise (0.7 m/sec, 30 min per day, five days per week), significantly increased the myoglobin concentration of the femorotibialis medius muscle in bar-headed geese as compared to nonexercised controls.
• 2.2. The myoglobin concentration differed among various muscles within a bird. The highest myoglobin concentrations were found in the primary flight muscle, the pectoralis major, and in cardiac muscle.
• 3.3. By physically conditioning their muscles, bar-headed geese may improve the oxygen flow to mitochondria and, thereby, enhance their ability to exercise under conditions of low oxygen partial pressures.

     

Myofibril and sarcoplasmic reticulum changes during muscle development: Activity vs inactivity

• 1.1. The purpose of this study was to determine whether biochemical changes of skeletal muscle that occur as a result of exercise in young rats persist into adulthood.
• 2.2. Littermates (10 days old) were assigned to a 3, 6 and 12 week control or training group. In addition, a rest-exercise group (R-E) and exercise-rest (E-R) group were included.
• 3.3. The rest-exercise and exercise-rest rats were maintained for the 12 weeks with the first 6 weeks being either rest or exercise and the condition reversed during the last 6 weeks of the experiment.
• 4.4. Myofibril ATPase activity of rat plantaris increased from the 10d to 12 week animals (P < 0.05). As anticipated, training resulted in a lowered activity at 6 and 12 weeks compared to controls.
• 5.5. The Ca²⁺ uptake and Ca²⁺-ATPase activity of the sarcoplasmic reticulum followed a similar pattern.
• 6.6. With regard to the exercise-rest rats, the myofibril and SR ATPase activities at 12 weeks were comparable to the 12 weeks control rats.
• 7.7. The rest-exercise group approximated the 12 week training group with regard to myofibril and SR ATPase activities (P > 0.05).
• 8.8. The results suggest that the training adaptations that occur during development of skeletal muscle return to normal, when training ceases in the adult rat.
• 9.9. Furthermore, animals that started to train prior to puberty do not have a greater capacity to adapt than animals which initiated training during adulthood.

     

Long-term starvation in Xenopus laevis daudin—III. Effects on enzymes in several tissues

• 1.1. Adult, female Xenopus laevis were subjected to 12 months of starvation.
• 2.2. Starvation resulted in a continuous reduction in the activity of both hepatic and renal glucose-6-phosphate dehydroganse.
• 3.3. Fructose-1,6-diphosphatase was significantly reduced at months 10 and 12 in the liver, and at months 4, 10, and 12 in the kidney.
• 4.4. Pyruvate kinase activity of muscle and liver decreased during the experimental period whereas the renal enzyme remained essentially unchanged.
• 5.5. Both hepatic and renal glutamate-pyruvate transaminase (GPT) and hepatic glutamate-oxaloacetate transaminase (GOT) showed a reduction of activity after 2 and 4 months of starvation followed by an increase in GPT but not in GOT.

     

Variations in tissue reserves,plasma metabolites and pancreatic hormones during fasting in immature carp (Cyprinus carpio)

• 1.1. Immature carp were subjected to 2-month fasting periods. Mobilization of reserves in liver and muscle, and the energy contribution of each reserve were studied. Changes in plasma glucose, amino acids, insulin and glucagon levels were determined throughout the experiment.
• 2.2. No changes were observed in plasma glucose, insulin or glucagon at 19 days of fasting, but plasma amino acids increased. At 50 days of fasting, both plasma glucagon and amino acids increased, liver glycogen decreased and muscle proteolysis began.
• 3.3. Between 50 and 67 days of fasting, plasma glucose and insulin decreased significantly, while glucagon and amino acids continued to increase. Strong muscular proteolysis was observed while liver glycogen stabilized.
• 4.4. The contribution of each reserve in liver and muscle to energy production throughout fasting is considered.

     

Changes in the activities of lysosomal enzymes in the fat body and midgut of two lepidopteran insects (Mamestra brassicae and Pieris brassicae) during metamorphosis

• 1.1. The activities of three lysosomal enzymes (acid phosphatase, β-galactosidase, catepsin D) was observed during metamorphosis in the fat body and midgut cells of two insects (Mamestra brassicae and Pieris brassicae).
• 2.2. The activities increased slightly during the feeding period and showed a sharp rise at the beginning of the wandering period.
• 3.3. Subsequently, a decrease was observed during the pre-pupal stage and pupation.
• 4.4. The activities increased again 2 days after the larval-pupal moult.
• 5.5. We suggest that an inhibitory mechanism works in the studied cells before pupation to protect the stored proteins from the degradation until the beginning of differentiation of imaginai cells in the pupal stage.

     

Deterioration of chum salmon (Oncorhynchus keta) muscle during spawning migration—III. Changes in protein composition and protease activity of juvenile chum salmon muscle upon treatment with sex steroids

• 1.1. Changes in protein composition and protease activity of juvenile chum salmon muscle upon treatment with sex steroids were investigated.
• 2.2. A slight breeding color was observed on chum salmon following the oral administration of 17α-methyltestosterone. Sarcoplasmic protein significantly decreased, while ninhydrin-positive substances from protein-free fractions significantly increased upon treatment with 17α-methyltestosterone. Autolytic activity of the fish treated with 17α-methyltestosterone drastically increased.
• 3.3. Estradiol-17β did not significantly influence the protein composition and autolytic activity.
• 4.4. These results indicate that androgen is closely related to the deterioration of chum salmon muscle.

     

Fiber type composition and enzyme activities of muscles in two models of obese rats

• 1.1. In oxidative (soleus) and glycolytic (extensor digitorum longus) muscles of obese Zucker rats, a significant decrease in the percentage of relative area occupied by glycolytic fibers was observed.
• 2.2. The activity of citrate synthase and β-hydroxy-acyl-CoA-dehydrogenase was significantly higher in muscles of obese than of lean Zucker rats.
• 3.3. In rats, 6 weeks after lesion of the ventromedial hypothalamus, no changes were observed.
• 4.4. This indicates that neither the proportion of oxidative fibers, nor the oxidative capacities are decreased in skeletal muscles of obese rats suggesting that insulin resistance cannot be ascribed to a higher glycolytic-oxidative fiber ratio.

     

Changes in chemical composition of muscle in young hybrids between russian sturgeon Acipenser guldenstadti brandt × beluga Huso huso l. (Pisces; acipenseriformes) under different levels of salinity

• 1.1. The salinity tolerance in young RS × B hybrids increases as the fingerlings grow. The specimens weighing about 7 g are able to tolerate the direct transfer to the water salinity 18%..
• 2.2. Under hypo- and iso-osmotic water ion concentration in the hybrid muscle free amino acids, the exchange of taurine for β-alanine and glycine takes place.
• 3.3. Under hyperosmotic conditions within the first 2 days in the hybrid muscle the water quantity declines, the protein quantity also slightly decreases, the urea and free amino acids concentration (mostly alanine, aspartic and glutamic acids, leucine), and a portion of reserved lipids increase.
• 4.4. During the next 4 days the muscle moisture, protein quantity, and the concentration of urea and free amino acids return to control values, but the portion of reserved lipids declines below the original level.

     

Correlation of water translocation,cuticle dehydration and post-ecdysial sclerotization by the american cockroach

• 1.1. Haemolymph volume decreases during the initial 16 hr post-ecdysial period, increases after water ingestion and subsequently drops until the inter-ecdysial level is reached.
• 2.2. Total body water follows a similar pattern, but the changes are not as pronounced.
• 3.3. Tissue water is inversely proportional to the total body water.
• 4.4. Soluble cuticle protein declines throughout the initial 16 hr period while both β-glucosidase and alkaline phosphatase activity is lost within 6 hr after ecdysis.
• 5.5. Dehydration of the cuticle also occurs during the immediate 6 hr post-ecdysial period.
• 6.6. These data suggest that the formation of the protein-insoluble matrix is linked with water loss.
• 7.7. Water removal may decrease the distance between molecules allowing specific reactions to take place.

     

Changes in high-energy phosphate metabolism in the water scorpion,Ranatra chinensis,under cold water-warm water stress

• 1.1. To evaluate changes in high-energy phosphate metabolism in the water scorpion (Ranatra chinensis) under restraint and cold water-warm water stresses, in vivo [³¹P]NMR spectra were obtained.
• 2.2. Under restraint stress, arginine phosphate (Arg-P) decreased by 10% after 1 hr and remained at that level thereafter, while β-ATP showed negligible changes over 6 hr.
• 3.3. As the water temperature gradually increased or decreased, the relative concentration of Arg-P decreased due to enzyme regulation.
• 4.4. Repeated cold water-warm water stress, which consisted of repeated 15 min exposures to cold water (5°C) followed by 15 min exposures to warm water (30°C) caused distinct decreases in Arg-P and β-ATP concentration. These decreases were dependent on the frequency of exposure.
• 5.5. Phosphomonoesters (PME) increased not only with restraint stress but also with cold water-warm water stress.
• 6.6. The effect of cold water-warm water stress on high-energy phosphate metabolism was greater than that of restraint stress.

     

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.

     

In vitro Ca deposition by rat matrix vesicles: Is the membrane association of alkaline phosphatase essential for matrix vesicle-mediated calcium deposition?

• 1.1. Phosphatidylinositol phospholipase C (PI-PLC) treatment of rachitic rat matrix vesicles (MVs) released about 80% of membrane-bound alkaline phosphatase (ALP), AMPase, PPiase into the media.
• 2.2. About 20% hydrolytic activity was not released from MV membranes by PI-PLC treatment.
• 3.3. SDS-polyacrylamide gel electrophoresis and Western blot analysis showed only one immunoreactive protein corresponding to the molecular weight of ALP present in the soluble fraction after PI-PLC treatment.
• 4.4. The specific activity of the released ALP was at least 5-fold higher than the residual activity.
• 5.5. After PI-PLC treatment, MVs also demonstrated an 80% reduction of AMP- or βGP-dependent calcium deposition.
• 6.6. The soluble fraction containing 80% of ALP activity was unable to support calcium deposition. The mixing of the soluble and insoluble fractions after PI-PLC treatment failed to fully restore calcium-depositing activity.

     

Copurification and separation of β-galactosidase and sialidase from porcine testis

• 1.1. A soluble sialidase was copurified apparently as an enzyme complex with acid β-galactosidase from porcine testis.
• 2.2. The sialidase exhibited its maximum activity at acidic pH. It was efficiently active towards 4-methylumbelliferyl-α-d-N-acetyl-neuraminic acid and sialyllactose, relatively inactive towards glycoproteins, and had little activity towards glycolipids.
• 3.3. The complex could be separated by sucrose gradient centrifugation or isoelectric focusing.
• 4.4. The separated enzymes had molecular weights about 600,000 for β-galactosidase and more than about 1,000,000 for sialidase by Sepharose 4B gel filtration.
• 5.5. SDS-polyacrylamide gel electrophoresis of the β-galactosidase showed three protein bands with molecular weights of 63,000, 31,000 and 20,000.

     

31P-NMR study on effects of electric shock on swimming leeches,Hirudo medicinalis

• 1.1. We evaluated the effect of electric shock on swimming leeches by measuring changes in high-energy phosphate metabolism using in vivo³¹P-NMR.
• 2.2. Leeches electrically stimulated during swimming showed anodal galvanotaxis, and stopped swimming with stimulation at strong current.
• 3.3. Comparison of the concentrations of high-energy phosphate metabolites before and after electric shock using ³¹P-NMR revealed a marked decrease in β-ATP and an increase in that of Pi.
• 4.4. Electric shock apparently induces excessive muscle fatigue in leeches, resulting in transient paralysis.