Occurrence of avidin-like biotin-binding capacity in various vertebrate tissues and its induction by tissue injury

• 1.1.|Avidin-like biotin-binding capacity was found in the liver and ovary of the common frog and in the kidney or lung of various avian species.
• 2.2. The induction of biotin-binding capacity by tissue injury was marked in various avian species, while in the lizard and in the skin of the common frog it was slight.
• 3.3. Avidin-like biotin-binding capacity or its induction was not found in the tissues of rainbow trout and three mammalian species studied.

     

Larval Musca domestica lipophorin biosynthesis

• 1.1. Larval Musca domestica lipophorin biosynthesis was studied in vitro.
• 2.2. The newly synthesized lipophorin has a density a little lower than the circulating lipophorin after 1 hr of incubation. After 3 hr of incubation the fat body cells transfer lipids to the lipophorin that attains the density of circulating lipophorin.
• 3.3. The lipophorin synthesized in vitro is identical to circulating lipophorin in density and in electrophoretical behavior.
• 4.4. However these two molecules must have differences since the circulating lipophorin transfers lipids to fat body cells while the synthesized in vitro does not.
• 5.5. The biosynthesis of Musca lipophorin shows differences with the Manduca sexta lipophorin biosynthesis.

     

Insulin,glucagon and catecholamine interactions in avian liver explants

• 1.1. A mechanical tissue chopper was used to obtain liver explants (35–75 mg) from 2- to 3-week-old chickens to determine both tissue sensitivity and metabolic effects of isoproterenol, avian insulin and glucagon.
• 2.2. Avian insulin had no effect on lipogenesis; however, lipogenesis was decreased by dibutyryl cyclic AMP. Insulin did not overcome a decrease in lipogenesis caused by catecholamines. Therefore, this control mechanisms is not modulated by insulin.
• 3.3. Preincubation in the presence of glucagon decreased in vitro lipogenesis. Preincubation in the presence of a 19–29 amino acid construct that approximated the radioimmune site for glucagon did not result in a similar effect. Therefore, this site does not relate to the biopotency of the hormone.
• 4.4. A previously noted catecholamine induced decrease in in vitro lipogenesis was verified, showing that points of in vitro regulation are under phosphorylation-dephosphorylation control.
• 5.5. Preincubation of slices (1 hr) with propranolol blocked the inhibition of lipogenesis caused by α and β adrenergic agonists (arterenol or isoproterenol) during a subsequent 2-hr incubation.
• 6.6. Preincubation of slices with either of these agonists decreased lipogenesis even following an extensive washout.
• 7.7. Inhibition could be overcome with propranolol, a β adrenergic antagonist.

     

Vitellogenesis in the shrimp,Penaeus vannamei: in vitro studies of the isolated hepatopancreas and ovary

• 1.1. Yolk proteins were isolated from ovaries of the shrimp Penaeus vannamei and used as an antigen for antibody production in rabbits.
• 2.2. Protein synthesis was measured for both the hepatopancreas and the ovary in vitro, and proteins present in both tissues were immunoreactive with the antibodies.
• 3.3. Extracts of shrimp eyestalks inhibited in vitro protein synthesis by both tissues. The inhibitory factor from the eyestalks was heat stable and had a molecular weight of 3300 daltons.

     

Specific biochemical and immunological properties of some water-soluble glycoproteins produced by rat gastric mucosal scrapings in vitro

• 1.1. Two major glycoprotein components were released in vitro by rat gastric mucosal cells after 4 hr incubation with radioisotopes: a high molecular weight fraction with the characteristics of a fucomucin and a low molecular weight fraction, the latter having a higher specific radioactivity than the former.
• 2.2. Pulse chase experiments indicate that several low and high molecular weight glycoproteins are synthesized simultaneously with no precursor-product relationship between them.
• 3.3. Common antigenic determinants, specific to the stomach were found on the 2 fractions, using immunofluorescence, both fractions appeared to be present in the same cells.

     

Comparison of the proliferation and differentiation of myogenic satellite cells and embryonic myoblasts derived from the turkey

• 1.1. Embryonic and posthatch turkey skeletal muscle development was compared in in vitro studies using clonal-derived embryonic myoblasts and satellite cells.
• 2.2. Although population doubling times were similar between the two lines (25.4 hr for satellite cells and 26.4 hr for embryonic myoblasts), embryonic myoblasts consistently began log phase growth 24 hr earlier than satellite cells.
• 3.3. Differentiation (fusion) of embryonic myoblasts was maximized by 36 hr in Dulbecco's Modified Eagle's Medium containing 1% horse serum compared with 72 hr for satellite cells.
• 4.4. When administered a serum-free medium which supports proliferation of turkey satellite cells, embryonic myoblasts differentiated to form myotubes.

     

Carbohydrate metabolism during osmoregulation in Chasmagnathus granulata dana, 1851 (crustacea,decapoda)

• 1.1. Since glucose is one of the main energetic substrates for general metabolic processes in crustaceans, analysis of carbohydrate levels can furnish information on the energy metabolism of intact animals during osmoregulation.
• 2.2. Different groups of Chasmagnathus granulata were transferred to different salinities (0 and 40%), and the glucose and glycogen concentrations in blood, gills, muscle and hepatopancreas were determined at the beginning of the experiment and 24, 72, 168 and 360 hr after the salinity changes.
• 3.3. Differences in tissues carbohydrate levels were observed between summer and winter, that reflected differences in reserve mobilization.
• 4.4. In the summer, hypo- and hyperosmotic shocks induced an increase in carbohydrate levels in almost all tissues studied, indicating gluconeogenesis.
• 5.5. In the winter, a carbohydrate mobilization occurred only in the gills and hepatopancreas after both osmotic shocks.
• 6.6. Thus, the substrate reserve used for energy production required for osmoregulation seems to be dependent on the season and tissues.

     

Characteristics of Crassostrea virginica crystalline style chitin digestion

• 1.1. Fundamental chitin digestion characteristics of Crassostrea virginica crystalline style were investigated.
• 2.2. Optimum temperature and pH were 34°C and 4.8. respectively.
• 3.3. The colloidal regenerated chitin (0.56mol/0.5 ml: GlcNAc equivalents) was saturating under all enzyme levels encountered.
• 4.4. There was no evidence of end product inhibition, even after 100 hr incubation.
• 5.5. Calculated K_m for the chitinase complex was 1.19mM when determined using a 30 min assay, but was only 0.70 mM when determined using a 4.6 hr assay.
• 6.6. Both K_m values are lower than reported for similar assays in other molluscs and for most bacteria.
• 7.7. Effect of substrate preparation on the kinetics are discussed.
• 8.8. Eight peaks of chitinase activity were resolved by DEAE-Fractogel ion exchange chromatography.

     

Characterization of the Melanoplus sanguinipes hemolymph after infection with Beauveria bassiana or wounding

• 1.1. The phenoloxidase activity, protein and carbohydrate levels were studied for 24 hr in the hemolymph of the migratory grasshopper, Melanoplus sanguinipes after artificial wounding of the insect cuticle or the injection of Beauveria bassiana conidia.
• 2.2. Injection or wounding induced a primary response and phenoloxidase activity was found to increase within 10–60 min. The values for phenoloxidase activity in viable B. bassiana-injected insects exhibited a secondary response, i.e., an increase 24 hr after injection.
• 3.3. In wounded insects and those injected with inactivated conidia, the phenoloxidase activity receded after the initial increase and remained at low levels.
• 4.4. Protein concentrations in the hemolymph increased immediately after infection and wounding and returned to basal levels during the course of the experiment.
• 5.5. Injection of viable B. bassiana resulted in a gradual increase in the protein concentrations between 12 and 24 hr.
• 6.6. There was no apparent change in the carbohydrate levels in either B. bassiana-infected or wounded insects.
• 7.7. These results are discussed in relation to their possible role(s) and interrelationships in the immune response to infection or wounding. Furthermore, we suggest that a “factor” is released after mechanical injury of the integument.

     

Lipid and carbohydrate metabolism of the intertidal crab Chasmagnathus granulata dana, 1851 (Crustacea: Decapoda) during emersion

• 1.1. Lipid, glucose and glycogen concentrations were measured in different tissues of the crab Chasmagnathus granulata during emersion.
• 2.2. After 6 hr of emersion no reduction in the total amount of carbohydrates was found to occur, suggesting that a general metabolic arrest was taking place.
• 3.3. A transitory increase in haemolymphatic glucose and lipid levels was observed. Possible causes are therefore discussed in relation to changes in the flux of substrates for energy production.
• 4.4. The mobilization of carbohydrates and lipids to the gills, observed only during summer, may be concerned with energy supplying for ionic regulation.

     

Active transport of d-glucose in intestinal segments,in vitro,of the scup,Stenotomus versicolor and the puffer,Spheroides maculatus

• 1.1. Active transport of d-glucose was shown using intestinal sac preparations, in vitro, made from two marine fish, the scup, Stenotomus versicolor and the puffer, Spheroides maculatus.
• 2.2. Differences in absorption characteristics were evident in populations from year to year.
• 3.3. Anaerobiotic conditions, i.e. 100 per cent nitrogen gassing of the incubation medium, inhibit the active transport of d-glucose in scup and puffer intestine.
• 4.4. Phlorizin, 5 × 10⁻⁴ M, inhibits the active transport of d-glucose in scup intestine.
• 5.5. Intestinal transmural glucose transport mechanisms operate well at incubation temperatures, 20°–27°C, i.e. temperatures close to habitat and holding tank temperatures, whereas movement of the sugar against a concentration gradient is interrupted at higher incubation temperatures, 29° and 30°C.
• 6.6. Detailed comparison of procedures and results with those used by other workers in the field of in vitro intestinal absorption of poikilotherms suggests that aerobic metabolism may not be a uniformly significant energy source in intestinal active transport.

     

Induction kinetics of immune antibacterial proteins in pupae of Galleria mellonella and Pieris brassicae

• 1.1. Pupae of Galleria mellonella and Pieris brassicae given an injection with live, non-pathogenic Enterobacter cloacae or abiotic foreign molecules induce an acquired immunity that corresponds with the synthesis of haemolymph proteins of antibacterial activity.
• 2.2. This humoral defensive response which persists for several days, differs quantitatively between insect species and between the inducers used, although very different foreign bodies induced the same immune proteins in both lepidopteran insects.
• 3.3. A stronger and longer lasting response was consistently noticed in pupae immunized with non-pathogenic bacterium than after sterile nutrient broth injections.
• 4.4. A demonstrably elevated activity of haemolymph lysozyme and trace activity of cecropins found in pupae of Galleria treated with saline W, a salt solution physiological to moths, disappear soon after 36 hr from injection.
• 5.5. In P. brassicae, however, sterile insect Ringer can give a varying, if present at all, immune response.
• 6.6. A mechanical injury (sterile wounding of insect body) can occasionally induce a similar but much weaker response.
• 7.7. The antibacterial activity was drastically reduced in Pieris or completely depressed in most pupae of Galleria when actinomycin D or cycloheximide was given at an early time post-immunization with E. cloacae.
• 8.8. It is concluded that the de novo synthesis of ribonucleic acid and immune proteins is required for expression of antibacterial activity in pupal haemolymphs.
• 9.9. The synthesis of an immune mRNA was completed about 7 hr after the injection of the immunizing bacteria.

     

Anaerobic metabolism of the common cockle,Cardium edule—IV. Time dependent changes of metabolites in the foot and gill tissue induced by anoxia and electrical stimulation

• 1.1. The levels of adenylates, arginine phosphate, arginine and various other metabolites related to anaerobic metabolism were determined in foot and gills of Cardium edule during anoxia (incubation in oxygen-free sea water) up to 48 hr and after electrical stimulation of the foot muscle.
• 2.2. In the foot the phosphagen was depleted quite rapidly in the first hours of anoxia. Free arginine levels rose while there was little octopine accumulation. d-Lactate appeared to be the predominant end product. Alanine and succinate were also produced and the concentration of aspartate decreased, suggesting this metabolite as precursor for succinate. In the gills, where no arginine phosphate was present, succinate production was higher than in the foot and exceeded lactate formation.
• 3.3. Only a small decline in ATP occurred during anoxia and consequently, only moderate changes in the energy charge were observed in both tissues.
• 4.4. Electrical stimulation of the foot also resulted in the breakdown of arginine phosphate and formation of mainly lactate and not octopine.
• 5.5. Total ATP consumption during anoxia has been calculated to be more rapid during the first hours. The ATP consumption rate was reduced by a factor of 7.8 during 48 hr of anoxia.
• 6.6. During the first 4 hr of anoxia about 60% of the total consumed energy (as ATP equivalents) was provided by the phosphagen, while later glycolysis contributed about 80%. During electrical stimulation about 75% of the energy supplied was derived from the phosphagen.
• 7.7. A comparison of the C. edule results with the data during anoxia and exercise of the jumping cockle is made.

     

Main kinetic characteristics of acetylcholinesterase from brain of Hypostomus punctatus,a Brazilian bentonic fish (cascudo)

• 1.1. A potentiometric method for the assay of cholinesterase has been proposed and compared with a colorimetric assay.
• 2.2. Main kinetic parameters of cholinesterase from Hypostomus punctatus brain were determined indicating that true acetylcholinesterase is by far the predominant enzyme in the brain of this fish.
• 3.3. We have compared our data with published results described from other fish species.
• 4.4. The enzyme inhibition achieved after 3 hr incubation of brain homogenates with ethyl-parathion have indicated that this enzyme shows a characteristic organophosphorous sensitive behavior.

     

Fate of ingested leucine and glucose in the sea star,Asterias rubens,during its annual reproductive cycle

• 1.1. The distribution of radiolabel from L-leucine [¹⁴C-UL] and D-glucose [¹⁴C-UL] was measured in the sea star Asterias rubens at 1, 6 and 24 hr after oral administration.
• 2.2. Incorporation of the label from both compounds was observed in pyloric caeca, coelomocytes and ovaries even after an incubation time of 1 hr.
• 3.3. Highest incorporation from both precursors was found in proteins, while substantial radioactivity was present in the amino acids, organic acids and neutral components. Lipids were hardly labelled from leucine and only slightly from glucose.
• 4.4. Radioactivity in proteins and lipids increased with increasing incubation time. No significant differences were found in the distribution patterns of radiolabel during the reproductive cycle.
• 5.5. The data obtained are discussed in terms of current knowledge on the translocation of nutrients in echinoderms.

     

In vitro non-enzymatic glycosylation of myofibrillar proteins

• 1.1. Glycation is non-enzymatic modification of proteins by sugars in which not only structural but also biological properties of proteins are altered.
• 2.2. Our in vitro experiments show that incubation of myofibrillar proteins with ribose results in sugar attachment to proteins and at the same time myofibrillar ATPase activity is lowered.
• 3.3. DETAPAC, aminoguanidine and 2-mercaptoethanol all partially block myofibrillar protein glycation and ATPase activity is less inactivated.
• 4.4. The dependence of ATPase activity of myofibrils incubated with ribose on the amount of 2-mercaptoethanol present suggests that also modification of SH groups is involved in enzyme inactivation.
• 5.5. Electrophoretic studies revealed that heavy chains of myosin, actin, and tropomyosins are proteins which are mainly glycated in vitro.

     

Absorption of glucose,glycine and palmitic acid by isolated midgut and hindgut from crickets

• 1.1. In vitro experiments indicated that midgut and hindgut anterior to the Malpighian tubules are important in absorption and processing of products of digestion in crickets.
• 2.2. Isolated hindguts from crickets (Gryllus assimilis, G. rubens and Scapteriscus acletus) absorbed and released into the incubation medium 20–30% of a load of [¹⁴C]glucose and 29–31% of a load of[¹⁴C]glycine.
• 3.3. Isolated midguts from the same crickets absorbed and released into the incubation medium 30–50% of the glucose and 43–52% of the glycine load.
• 4.4. Radiolabelled palmitate was absorbed into epithelial cells of isolated mid- and hindguts, but little was transported and released into the incubation medium.

     

In vivo and in vitro synthesis of some serum peptides

• 1.1. A high amount of [¹⁴C]leucine is incorporated in vivo into rat serum peptides after 1 hr of isotope administration.
• 2.2. In vitro [¹⁴C]leucine appears in reconstituted blood plasma and in rat liver peptide mixtures. After 4hr of perfusion the radioactivities amount to 2078 ± 913 and 2120 ± 549 cpm/mg of peptides, respectively.
• 3.3. The serum and liver peptides analysed show a great aggregation ability and so it is difficult to decide which of the liver peptides are destined for the serum.

     

Evoked effects of oestradiol on hepatic cholesterol 7α-hydroxylase and drug oxidase in castrated rats

• 1.1. Oestradiol administration in castrated rats resulted in an increased activity of the cholesterolα-hydroxylase and a decreased activity of the drug oxidase enzyme systems.
• 2.2. Aqueous solutions of oestradiol (up to 25·10⁻⁶M) incubated in vitro with microsomes, binds into the microsomal membrane framework reducing the activity of both enzyme systems.
• 3.3. The specific activity of cholesterol 7α-hydroxylase. drops after 3 hr preincubation with oestradiol to at least 70% of its original value.
• 4.4. Actinomycin D and cycloheximide administration reduced the oestradiol-induced and control cholesterol 7α-hydroxylase activity to the same level, 6 hr after the injections.

     

Influence of dietary sources of fat on lipid synthesis in mink (Mustela vison) mammary tissue

• 1.1. The fatty acid composition of the triglyceride fraction of mink milk sampled during mid-lactation (day 28 post partum) from two nursing mink was compared to that of plasma samples and to the fatty acid composition of the feed rations used.
• 2.2. Chemical analysis of the triglyceride composition of mink milk demonstrated only minute concentrations of fatty acids with a chain length below C₁₄.
• 3.3. The saturated C_16:0- and C_18:0-unit fatty acids in mink milk made up for 24–40% of the total amount of fatty acids extracted, the remainder being represented by mono and polyunsaturated long-chain (C₁₆-C₂₄) fatty acids.
• 4.4. Preliminary in vitro experiments proved the incorporation of¹⁴C-labelled glucose, acetate or palmitate into triacylglycerols in cultures of mink mammary tissue to be linear for at least 2 hr.
• 5.5. The in vitro capacity for de novo fatty acid synthesis in mink mammary tissue using 14C-labelled glucose or acetate was low, i.e. ranging from 0.096–0.109 nmol/g (fresh tissue)/min, and amounted to only about 5% of that obtained in the case of [¹⁴C]palmitic acid incubation.
• 6.6. Following ¹⁴C-labeIled acetic or palmitic acid incubation of mink mammary tissue neither desaturation nor chain elongation was observed.
• 7.7. In response to long-term feeding on rations with two different sources of animal fat (F = fish oil or L = lard) the influence of compositional changes in dietary neutral lipids on the fatty acid composition of the lipids of mink milk is discussed.