Oxidative metabolism of the inner and outer ventricular layers of carp heart (Cyprinus carpio L.)

• 1.1. The biochemical characteristics of homogenates and mitochondria isolated from the outer and inner layers of the ventricular myocardium of carp were studied.
• 2.2. The homogenate prepared from the inner layer exhibited higher activity of cytochrome oxidase than that from the outer layer. No difference was found in the activity of cytochrome oxidase between mitochondria from the inner and outer layers. Difference spectra of cytochromes also showed that their content in mitochondria of both layers is similar and that the higher oxidative capacity of the spongious layer is due to a higher content of mitochondria.
• 3.3. In comparison with rat heart a higher content of cyt aa₃ and a lower content of Cyt b and cyt cc₁ were found in carp heart mitochondria.
• 4.4. In comparison with rat heart, carp heart mitochondrial enzymes were more sensitive to freezing-thawing and to detergent action.

     

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.

     

DNase-I-like enzyme from the carp liver—Inhibition by muscle and endogenous actin

• 1.1. DNase-I-like activity occurs in the carp (Cyprinus carpio) liver cytosol (supernatant 105,000g).
• 2.2. The enzyme resembles DNase I from bovine pancreas in respect to the molecular mass (~31 kDa), pH (7.4) and ion requirements (Mg²⁺, Ca²⁺) and the ability to degrade native as well as denatured DNA.
• 3.3. As judged by comparison of DNase zymograms obtained after native- and SDS-PAGE, the enzyme occurs in the three molecular forms of similar molecular weight and different charges.
• 4.4. All these forms are inhibited by rabbit skeletal muscle actin as well as by endogenous actin isolated from the carp liver cytosol.
• 5.5. DNase from the carp liver cytosol does not interact with the antibodies directed against DNase I from bovine pancreas and against DNase I from the rat and bovine parotid glands.

     

Troponin C of the Antarctic icefish (Champsocephalus gunnari) white muscle

• 1.1. The troponin C (TN-C) from the Antarctic icefish (_{Champsocephalus gunnari}) has been isolated to homogeneity by a procedure involving extraction from acetone powder, DEAE-Sepharose 4B and AcA 54 column chromatography.
• 2.2. The calcium-induced conformational changes of apo TN-C have been studied by absorption difference spectroscopy, circular dichroism and intrinsic fluorescence.
• 3.3. The results indicate that the overall characteristics of icefish TN-C (such as amino acid composition, modifications of helix content and of the microenvironment of aromatic residues as a function of calcium binding) are quite similar to those of rabbit TN-C.
• 4.4. The intrinsic fluorescence properties are close to those reported for pike and carp TN-C.

     

Oral administration of insulin in winter-acclimatized carp (Cyprinus carpio) induces hepatic ultrastructural changes

• 1.1. The intestinal absorption of insulin in carps was assessed examining the transepithelial passage of ingested gold-labeled hormone by electron microscopy. Insulin transfer occurred mainly through the intercellular spaces between the enterocytes.
• 2.2. When reaching the lamina propria, the gold-labeled hormone gathered predominantly around the granules of the granular cells, and therefore can enter the circulatory system via the blood capillaries which are found in close contact with these cells.
• 3.3. Winter-acclimatized carp were also capable of internalizing the hormone when fed with insulin.
• 4.4. Furthermore, the absorbed hormone revealed full activity in regard to the observed changes in the ultrastructure of the liver cells of the treated cold-adapted fish.
• 5.5. The fish ingesting the hormone underwent the same type of hepatic ultrastructure reprogramming observed when winter-acclimatized carps are injected intraperitoneally with insulin, i.e. conversion to a phenotype corresponding to hepatocytes from summer-adapted carp.
• 6.6. The oral absorption of insulin by winter-acclimatized fish and its effect in reversing the cold-adaptive state might be useful for the fish culturing industry.

     

Renal response to hypoxia in carp,Cyprinus carpio: Changes in glomerular filtration rate,urine and blood properties and plasma catecholamines of carp exposed to hypoxic conditions

• 1.1. Changes in glomerular nitration rate (GFR), urine and blood properties and plasma catecholamines of carp were investigated during and following hypoxia.
• 2.2. GFR and urine flow decreased with increased urinary concentrations of bio-components, except protein, in the course of hypoxia.
• 3.3. Decreases in blood pH, and increases in haematocrit value and plasma K⁺, Ca²⁺, Mg²⁺, inorganic phosphate (P_i), ammonia, lactic acid and catecholamines (CAs) were observed as hypoxia progressed.
• 4.4. Increased GFR and urine flow, and higher values for urinary components, except protein, compared with those of the control were found in the initial post-stress stage.
• 5.5. The possible significance of increased plasma CAs in relation to changes in renal function in hypoxic carp is discussed.

     

Characterization of two distinct latent proteinases associated with myofibrils of crucian carp (Carassius auratus cuvieri)

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• 1.1. Enzymatic properties of two distinct proteinases tightly associated with crucian carp myofibrils were characterized.
• 2.2. These proteinases were latent but activated at 50 and 60°C, respectively.
• 3.3. The optimum pH of 50°C-proteinase was neutral-alkaline, while that of 60°C-proteinase was weak acid-neutral pH.
• 4.4. Both proteinases required more than 1% NaCl for the activity, but 50°C-proteinase was partially inhibited at higher concentrations of NaCl.
• 5.5. Both proteinases were regarded as trypsin-like proteinases belonging to a serine proteinase family, but only 60°C-proteinase was sensitive to urea, n-butanol and iso-propanol.

     

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Clastogenic effects of five carcinogenicmutagenic chemicals on the cells of the common carp,cyprinus carpio L.

• 1.1. The cytogenetic effects of various i.p. treatments with five carcinogenic-mutagenic chemicals and three doses for each (aflatoxin Bl, Aroclor 1254, benzidine, benzo[a]pyrene and 20-methylcholanthrene), were investigated in the cells of the common carp, Cyprinus carpio.
• 2.2. Injection with distilled water and corn oil served as the two control groups.
• 3.3. For detecting cytogenetic damage we used two test systems, chromosomal aberrations (CA) in kidney cells and micronucleated erythrocytes (M).
• 4.4. At 48 hr after treatment with the chemicals under investigation, the frequencies of CA and M were clearly increased in a dose-response manner compared to the control groups.

     

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The effect of water loss upon the urate,urea and ammonia content of the egg of the Japanese quail Coturnix coturnix japonica

• 1.1. The urate, urea and ammonia content of the whole egg of the Japanese quail was measured in late incubation in eggs subject to different rates of water loss.
• 2.2. High rates of water loss substantially increased egg urate content, but had little or no effect on urea or ammonia content.
• 3.3. Allopurinol, an inhibitor of urate synthesis, reduced egg urate content to low levels, but produced no effect on urea content, and a small reduction in ammonia content.
• 4.4. The urea concentration of the embryo was lower than in allantoic fluid.
• 5.5. It is concluded that urate production by the avian embryo is primarily concerned with the modification of allantoic fluid composition.

     

The energetics of shedding: Energy content of snake skin

• 1.1. The weight and energy content of sloughed skins of 92 individual snakes of 22 different species in three families were measured.
• 2.2. Weight and total energy content of shed skins were highly correlated with body weight.
• 3.3. The heat of combustion (kJ/g) of sloughed skins varied significantly among families and was higher in species having unkeeled scales than in those with keeled scales.
• 4.4. The presence of keels significantly affected weight of skins, even when skin weight is adjusted for covariance with body weight.
• 5.5. Neither body weight nor ambient temperature significantly affected the heat of combustion of sloughed skins.
• 6.6. The energy content of shed skin, expressed as a proportion of daily metabolism, decreased with ambient temperature, but the effect is minimized in large snakes.
• 7.7. Small snakes expended relatively less energy in sloughed skins than large snakes when the expenditure is expressed in terms of total daily metabolized energy.

     

The effect of dietary protein level and ration level on excretion of ammonia in common carp,Cyprinus carpio

• 1.1. Nitrogenous excretion in the form of ammonia was determined in common carp of 65.0 ± 8.0 g body weight in metabolism chambers. The fish were fed with 20, 35 and 50% dietary protein at 1, 2 and 3% body weight per day ration level.
• 2.2. Nitrogenous excretion as a percentage of ingested food increased with an increase of dietary protein but decreased with an increase of ration level.
• 3.3. The energy lost in excretion ranged from 4.19% with 20% dietary protein at 3% ration level to 8.74% with 50% dietary at 1% ration level.

     

Carbonic anhydrase III content in various equine muscles

• 1.1. In this study, carbonic anhydrase III (CA-III) content in 18 equine muscles was determined by enzyme immunoassay.
• 2.2. It was found to differ in several muscles.
• 3.3. That in external intercostal muscle, rectus abdominis muscle and splenius muscle from four horses was very high.
• 4.4. Although the masseter muscle had only type I fibers, CA-III content was similar to that in mixed-fiber type muscles such as the biceps femoris muscle.
• 5.5. It thus appear that equine type I fibers can be further subgrouped.

     

Specific dynamic action and feeding metabolism in common carp,Cyprinus carpio L.

• 1.1. Oxygen uptake attributable to Specific Dynamic Action (SDA) was measured in common carp, Cyprinus carpio L. (63.6–84.0 g) fed on 20, 35 and 50% dietary protein at 0.40 to 1.00% ration levels at 28°C.
• 2.2. After feeding both SDA magnitude and mean peak oxygen consumption increased directly with dietary protein and ration levels. SDA duration was not significantly related to dietary protein but significantly increased with ration levels.
• 3.3. SDA coefficients were 8.99, 13.51 and 15.94% with 20, 35 and 50% dietary protein showing a direction relationship to the protein content. The SDA coefficient did not change with ration size.
• 4.4. SDA models resulting from this work are of great interest for the aquaculturist, as post-feeding oxygen requirements in an intensive fish culture can be predicted where dietary protein and ration levels are known.

     

Thermal stability of fish myosin

• 1.1. Thermal stability of fish myosin has been studied by using differential scanning calorimetry (DSC) and circular dichroism (CD).
• 2.2. The temperature range of the sharp decrease in α-helical content agreed very closely with that of the endothermic peaks.
• 3.3. There was a high correlation between the enthalpy of denaturation (ΔH) and the decreasing quantity in α-helicity (Δh).
• 4.4. The structure of fish myosins was much more unstable than that of rabbit.
• 5.5. The instability of fish myosins was reflected in its rod moiety.

     

The glycogen content in stressed marine bivalves: The initial absence of a decrease

• 1.1. Changes in the glycogen content, condition, stomach content and acetic acid concentration of mussels Mytilus edulis and cockles Cerastoderma edule were followed during periods of up to 14 days of exposure (to air) at temperatures of 5 and 20°C.
• 2.2. In animals with a high glycogen content the glycogen is not used during the first 3 to 7 days, at high and low temperature respectively.
• 3.3. After this latent period the glycogen concentration often decreased, coinciding with a high mortality and an increase of the concentration of acetic acid.
• 4.4. In cockles with a low glycogen content, and kept at a high temperature, glycogen can be used from the beginning of the stress period.
• 5.5. Between species no clear differences were found.
• 6.6. The stomach content decreased during exposure; however, the stomach content amounted to only 0.5 to 0.7% of the body weight, and is thought to be of minor importance as an energy source during the stress period.
• 7.7. Especially at the higher temperatures glycogen finally is transformed into acetic acid.
• 8.8. It is concluded that during exposure, the animals do not die because of a lack of energy reserves, but because of a high accumulation of acids.

     

Sample Preparation Scale-Up for Deep N-glycomic Analysis of Human Serum by Capillary Electrophoresis and CE-ESI-MS*

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Highlights

• •Efficient sample preparation workflow for deep N-glycomics analysis from serum.
• •Temperature gradient denaturing protocol to prevent protein precipitation.
• •Decrease of free sugar content in serum enhanced PNGase F digestion efficiency.
• •Modified evaporative labeling method increased fluorophore labeling yield.