Changes of the fatty acid composition in smolts of masu salmon (Oncorhynchus masou), associated with desmoltification and sea-water transfer

• 1.1. Tissue lipid compositions of desmoltified yearlings of masu salmon (Oncorhynchus masou) obtained by keeping smoltified fish in fresh water, were examined and compared to those of smoltified fish before and after transfer to sea-water (SW).
• 2.2. Lipid contents of muscle, liver, gut and gills of desmolts tended to increase compared to those of initial smolts.
• 3.3. The increased proportion of triacylglycerol (TG) and decreased proportion of phospholipids (PL) characterized the tissue lipids of desmolts.
• 4.4. Liver and muscle lipids showed no distinct differences both in content and proportion between initial and SW smolts, but gut and gill lipids of SW smolts decreased in content accompanied by a decrease of TG and an increase of PL in proportion.
• 5.5. Excepting muscle non-polar lipids, tissue lipids of desmolts contained more mono-unsaturated fatty acids and saturated fatty acids and less polyunsaturated fatty acids (PUFA), especially (n-3) PUFA such as 22:6(n-3), than those of initial and SW smolts.
• 6.6. No large differences in fatty acid composition were seen between initial and SW smolts except for the gut.
• 7.7. The proportion of (n-3) PUFA in the gut of SW smolts was higher than that of initial smolts.
• 8.8. The results indicated that masu salmon smolts can modify their lipid metabolism to adapt to ambient salinity changes. The proportion of (n-3) PUFA particularly in polar lipids, or in osmoregulatory organs such as gut and gills, was seen to be critical in lipid types of freshwater- or sea-water-adapted fish.

     

The influence of calcium and magnesium on sea bass (Dicentrarchus labrax) intestinal brush border membrane purification and activity

• 1.1. The activity of brush border enzymes (alkaline phosphatase, maltase, sucrase, trehalase, leucine amino peptidase) was higher in purified membranes prepared with calcium. The contamination of these membranes with basolateral membranes was also lower (1.27 for Na-K-ATPase activity ratio).
• 2.2. The extraction of brush border lipids was carried out according to Folch adapted method. Two dimensional thin layer chromatography was used to separate the phospholipidic fractions. Fatty acids of phospholipids were analysed using gas chromatography after acid transmethylation (column SP 2330).
• 3.3. Phospholipids are composed of phosphatidylcholine (PC: 33%), phosphatidylethanolamine (PE: 30%), sphingomyeline (SM: 21%), phosphatidylserine (PS: 14%) and phosphatidylinositol (PI: 2%). 4. PC, PE and PS are characterized by high levels of unsaturated fatty acids (monounsaturated MUFA: 21.5% and polyunsaturated PUFA: 34.9%). The most abundant PUFA belong to the (n-3) family [18:3 (n-3), 20:5 (n-3) and 22:6 (n-3)].
• 4.5. Fatty acids from sphingomyelin of purified membranes have low proportions of PUFA (13.5%) but higher proportions of MUFA (39.5%).
• 5.6. No specific differences were found between calcium and magnesium prepared membranes.
• 6.7. The low content in LPC and the absence of LPE confirmed the absence of major structural lipids transformation during the membrane purification with calcium or magnesium.
• 7.8. Glycine transport was measured during 10 sec at different temperatures using the rapid filtration technique. Glycine transport was higher with Na⁺ than with K⁺. In the presence of Na⁺, this transport increases with temperature.
• 8.9. Arrhenius curves were mono phasic without obvious breakpoint and indicated no phase transition in the lipid bilayer.
• 9.10. A significant Na⁺ dependent glycine transport has been characterized at low temperatures (0°C) which suggests a possible role of membrane polyunsaturated fatty acids in the control of glycine transport.

     

Selective effect of melatonin on the proliferation of lymphoid cells

• 1.1. The present study was designed to investigate the effect of melatonin on the proliferation of normal lymphocytes and certain T-lymphomas and myelomas under in vitro conditions.
• 2.2. The results revealed that administration of 200 μM melatonin inhibited significantly the incorporation of [³H]thymidine into both normal mouse and human lymphocytes and T-lymphoblastoid cell lines.
• 3.3. On the contrary, melatonin provoked an increase of myeloma cell proliferation.
• 4.4. The influence of melatonin on hybridoma cell lines was negligible.
• 5.5. Collectively, these data demonstrated that the chief pineal indole affect selectively the processes of lymphoblastoid cell growth.

     

Comparative study of the endemic freshwater fauna of lake baikal—I. Phospholipid and fatty acid composition of two mollusc species,Baicalia oviformus and Benedictia baicalensis

• 1.1. Lipid and phospholipid compositions of endemic freshwater molluscs belonging to the class Gastropoda, Baicalia oviformus and Benedictia baicalensis, were studied.
• 2.2. The fatty acids composition of total lipids, neutral, glyco- and phospholipid fraction was investigated by capillary gas chromatography-mass spectrometry.
• 3.3. Ninety-five fatty acids were identified: 23 saturated (both iso- and anteiso-), 28 monoenoic, 14 dienoic and 30 polyenoic.
• 4.4. High percentage of the two main acids, 18:4 and 18:4(n-3) in phospholipid and glycolipid fractions were identified.
• 5.5. A number of unusual polyunsaturated fatty acids, such as 19:4, 18:5(n-3), 24:4(n-6), 24:5(n-6), 24:6(n-3), and furanoid acids, were found.

     

Combined effect of adenosine,alpha adrenergic and adenosine antagonists on serum insulin and insulin secretion from rat pancreatic islets

• 1.1. The effect of adenosine separately or in combination with alpha-1 adrenergic antagonist prazosin and alpha-2 adrenergic antagonist yohimbine as well as adenosine antagonists 8-phenyltheophylline and xanthine amine conjugate on glucose-induced insulin secretion from isolated rat pancreatic islets was studied.
• 2.2. Their in vivo effects on serum glucose and insulin levels were also investigated. Adenosine at 10 and 100 μM inhibited significantly, insulin secretion from the isolated islets whereas at 10 mM slightly increased the secretion of insulin.
• 3.3. Prazosin used at 100 μM inhibited insulin secretion. When it combined with adenosine (10 μM) it augmented the inhibitory effect of adenosine.
• 4.4. In vivo prazosin (21 mg/kg bodywt) caused a hyperglycaemia which was accompanied by hypoinsulinaemia.
• 5.5. Concurrent administration of this drug with adenosine neither affect the hyperglycaemic nor the hypoinsulinaemic effects of adenosine.
• 6.6. On the other hand, yohimbine (100 μM) has no effect neither separately nor in combination with adenosine (10 μM) in modulating the inhibitory effect of adenosine on insulin secretion.
• 7.7. When Yohimbine administered at 19.5 mg/kg body wt it did not alter serum glucose but it markedly increased the serum insulin level. Its combined administration with adenosine reduced the hyperglycaemic effect of adenosine with a remarkable increase in serum insulin.
• 8.8. Both adenosine-antagonists were ineffective in alteration of insulin secretion.
• 9.9. However, combination of 8-phenyltheophylline with adenosine (10 μM) totally blocked the inhibitory effect of adenosine on insulin secretion while xanthine amine conjugate failed to prevent this effect of adenosine.
• 10.10. These results indicate that the inhibitory effect of adenosine on insulin secretion is neither mediated via alpha-1 nor alpha-2 adrenoceptors. It might be via activation of specific adenosine receptors on rat islets which are sensitive to blockade by 8-phenyltheophylline.

     

Serotonin,histamine and somatostatin modulation of PMA-stimulated phosphorylation of 130kDa Ca2+ pump-like protein from rat cerebellum synaptosomal membranes

• 1.1. Influence of some neurotransmitters and neuromodulators on the PMA-stimulated phosphorylation in vitro of calcium pump-like protein from rat cerebellum synaptosomal membranes was examined.
• 2.2. The prolonged time (up to 6 min) of synaptosomal membranes preincubation with 1 and 10 μM serotonin results in the increase of phosphorylation. The decrease of phosphorylation up to 80% of control value was observed for 100 μM serotonin.
• 3.3. The most stimulating effect on 130kDa protein phosphorylation was observed with 1μM of histamine (160% of control value).
• 4.4. 1 and 0.1 μM somatostatin triggered a short-time transient increase of 130 kDa phosphorylation (up to 135% of control value).

     

Partial purification of rat liver cytoplasmic acetyl-CoA synthetase; Characterization of some properties

• 1.1. Rat liver cytoplasmic acetyl-CoA synthetase was partially purified (purification factor = 23, yield = 30%).
• 2.2. The apparent K_ms for acetate, coenzyme A, ATP and MgCl₂ were determined and found to be 52.5 μM, 50.5 μM, 570 μM and 1.5 mM, respectively.
• 3.3. The partially-purified enzyme showed a low affinity for short-chain carbon substrates other than acetate.
• 4.4. The properties of the partially-purified enzyme were compared with those of enzymes from other sources.

     

Comparison of muscle lipid composition between marine and landlocked forms of sockeye salmon (Oncorhynchus nerka)

• 1.1. Total lipid content, and lipid class and fatty acid compositions in the muscle were analyzed for marine and landlocked forms of sockeye salmon.
• 2.2. Little difference was found for the total lipid content in the muscle between both forms.
• 3.3. Triglycerides were higher in the marine form than those in the landlocked one, but phospholipids showed an opposite tendency.
• 4.4. In the fatty acid composition of total lipid, percentages of 20:1 and 22: 6n-3 were higher in the marine form, while 18:2, 18:3n-3, 18:4n-3 and 20:4n-3 were more abundant in the landlocked one. Fairly high levels of 22:1 were present only in the marine form.

     

Ostrich fructose-1,6-bisphosphatase: Kinetic characterization of the liver isoenzyme

• 1.1. Purified ostrich (Struthio camelus) liver fructose-1,6-bisphosphatase exhibited an absolute requirement for Mg²⁺.
• 2.2. The enzyme catalyzed the hydrolysis of fructose-1,6-bisphosphate, sedoheptulose-l,7-bisphosphate and ribulose-l,5-bisphosphate.
• 3.3. S_0.5 for substrate was 1.4 μM.
• 4.4. AMP was a potent non-competitive inhibitor with respect to substrate (K_i of 25 μM).
• 5.5. Fructose-2,6-bisphosphate was a potent competitive inhibitor of the enzyme (K_i of 4.8 μM).

     

Effects of recombinant salmon growth hormone on hypophysectomized male Fundulus heteroclitus

• 1.1. Recombinant salmon growth hormone at doses of 0.8 and 2.1 μg/g significantly enhanced linear growth in hypophysectomized male killifish, Fundulus heteroclitus, over that of controls and a significant regression was found between growth and the logarithm of dose.
• 2.2. Bovine growth hormone elicited significant growth enhancement at all three dosages tested (1,4 and 10 μg/g) and a significant log/dose relationship was also observed.
• 3.3. Observations on the relative weight of the gonads indicate that whole salmon pituitary extract (25 μg/g) possesses strong gonadotropic activity and that both bGH and rsGH had smaller but significant effects on the gonads.
• 4.4. It is suggested that growth hormone may play a subsidiary synergistic role to other pituitary hormones in gonadal development.

     

Identification and characterization of a third form (D3) of cyclic 3'5'-nucleotide phosphodiesterase from rhizobwm fredii

• 1.1. A third form (D3) of cyclic nucleotide phosphodiesterase from Rhizobiumfrediiv/as detected and characterized for the first time.
• 2.2. The enzyme could hydrolyse both cyclic AMP and cyclic GMP with apparent K_m for cyclic AMP of approx. 0.2 μM.
• 3.3. D3 cyclic nucleotide phosphodiesterase had a pH optimum of about 6.0 when hydrolysing cyclic AMP.
• 4.4. The enzyme lost almost all its activity when heated to 60°C for 20 min.
• 5.5. Gel filtration with Sephadex G-100 gave a mol. wt of approx. 42.5 kD for the native enzyme.

     

Poly(d(A—T)) dependent RNA polymerase activity after treatment of nuclei with micrococcus nuclease

• 1.1. Rat liver nuclei were incubated with or without 20 units micrococcus nuclease (EC3.1.4.7)/mg nuclear DNA.
• 2.2. The soluble poly(d(A—T)) dependent RNA polymerases were reduced in activity to 15–20% that of the controls after treatment with micrococcus nuclease.
• 3.3. RNA polymerases I plus III activities were completely, RNA polymerase II activity partially reversible on removal of the DNA released into the soluble fraction by treatment of nuclei with micrococcus nuclease.
• 4.4. Inhibitory constants obtained with the solubilized DNA were 17.1 μM and 20.7 μM nucleotide-DNA for RNA polymerases I plus III and RNA polymerase II, respectively. The corresponding inhibitory constants obtained with native salmon DNA were 23.0 μM and 34.4 μM nucleotide-DNA.

     

Ionic dependence and vanadate sensitivity of (Na+, K+)-ATPase isolated from branchial sac of ascidians

• 1.1. Ion dependence and vanadium-induced inhibition on branchial sac ATPase in five species of ascidian Phlebobranchiata (vanadium-accumulating) and Stolidobranchiata (iron-accumulating) were studied.
• 2.2. The ATPase was obtained from the microsomal fraction, which was prepared from each ascidian branchial sac.
• 3.3. The ATPase was dependent on Mg²⁺ and activated by exogenous Na⁺ + K⁺.
• 4.4. Ouabain inhibited the ATPase activity in vitro, 10 μM to 100 μM vanadate, in vitro, suppressed the (Na⁺, K⁺)-ATPase.

     

Interrelationships between gluconeogenesis and uricogenesis in chicken liver

• 1.1. Experiments performed on isolated hepatocytes and perfused liver of starved chickens showed that gluconeogenesis from lactate, glycerol and fructose was inhibited by 22–100% on addition of urate precursors.
• 2.2. The inhibition was associated with an increased rate of urate formation.
• 3.3. 2,4-Dinitrophenol (40 μM), 2-bromooctanoate (2 mM) and 3-mercaptopicolinate (3MPA) (0.5 mM) were inhibitory with respect to gluconeogenesis but did not significantly affect the rate of urate formation.
• 4.4. The possible interrelationships between gluconeogenesis and uricogenesis are considered in terms of a competition for ATP and for other metabolites between the two pathways.
• 5.5. An interplay of both pathways at the level of anion transfer across the inner mitochondrial membrane is also discussed.

     

Characterization and purification of biliverdin reductase from the liver of eel,Anguilla japonica

• 1.1. Biliverdin reductase from the liver of eel, Anguilla japonica was characterized and purified with a novel enzymatic staining method on polyacrylamide electrophoretic gel.
• 2.2. This enzyme could use both NADPH and NADH as coenzyme. The K_m of NADPH was 5.2 μM, while that of NADH was 5.50 μM.
• 3.3. The optimum reaction pH for using HADPH as coenzyme was 5.3. That for NADH was 6.1. The optimum reaction temperature is 37°C.
• 4.4. When NADPH was used as coenzyme, the K_m of biliverdin was 0.6 μM. When NADH was used as coenzyme, the K_m of biliverdin was 7.0 μM.
• 5.5. The activity of the enzyme was inhibited by the concentration of biliverdin. Also, the potency of the enzyme was much less than that of the analogous enzyme isolated from mammals.
• 6.6. This is a fairly stable enzyme with a mol. wt around 67,000. Its estimated pI was pH 3.5–4.0.
• 7.7. This is the first time biliverdin reductase has been isolated and characterized from a vertebrate other than mammals. The property of it is quite different from that of mammals.

     

Effects of phosphatidylserine on the oxidation of low density lipoprotein

• 1.1. LDL was incubated in the presence of 1 μ M CuSO₄ for 18 hr at 37°C. The content of lipoperoxides was found to be approx. 40 nmol MDA equivalents/mg LDL protein. The addition of 50 μM phosphatidylserine (PS) reduced the content of lipoperoxides to 15% of control values.
• 2.2. The electrophoretic mobility observed for LDL oxidized in the presence of PS approximated the mobility observed for native LDL.
• 3.3. The formation of conjugated dienes was strongly inhibited when LDL was oxidized in the presence of PS.
• 4.4. The addition of 50 μM phosphatidylcholine, phosphatidylglycerol and cardiolipin did not alter the extent of LDL oxidation.
• 5.5. PS did not inhibit the oxidation of LDL mediated by J774 macrophages in the presence of Ham's F-10 culture medium. Under these conditions, PS was found to be an excellent substrate for oxidation.

     

Human GMP synthetase

• 1.1. The specific activity of GMP synthetase was measured in several human tissues and found to be highest in cultured skin fibroblasts, followed by bone marrow, leukocytes, erythrocytes. placenta, and liver.
• 2.2. The enzyme from fibroblasts was purified approximately 50-fold by ammonium sulfate fractionation and gel filtration.
• 3.3. The K_m values were determined to be 4.9μM for XMP, 270μM for ATP. and 340 μM for glutamine.
• 4.4. Ammonium sulfate could replace glutamine as the amino donor but was much less efficient.
• 5.5. The enzyme was specific for ATP as the energy source.
• 6.6. Unlike the calf thymus enzyme, the human enzyme has no requirement for a reduced sulfhydryl compound.
• 7.7. Human GMP synthetase is inhibited by ATP, dATP, azaserine, and hydroxylamine.

     

Copper toxicity and adaptation in the marine diatom Ditylum brightwellii

• 1.1. The planktonic diatom Ditylum brightwellii, grown in light-limited resp. nitrogen-limited continuous culture, has been exposed to Cu levels, comparable with those in the Scheldt estuary.
• 2.2. At increasing levels D. brightwellii initially detoxified Cu, producing metal-binding ligands (amino acids), and increasing its cell volume.
• 3.3. In light-limited D. brightwellii, photosynthesis could be adjusted to increasing Cu stress, division rates remained constant, and cells proved to be adaptable to 200 nM dissolved Cu.
• 4.4. Nitrogen-limited D. brightwellii detoxified Cu inadequately: it stored large amounts of Cu (30–60 μM) that inhibited cell division.

     

Inhibition of dihydrofolate reductase by palmitoyl coenzyme A

• 1.1. Palmitoyl-CoA was found to inhibit bovine liver dihydrofolate reductase.
• 2.2. 50% inhibition of the enzyme was obtained with 1.5 μM palmitoyl-CoA.
• 3.3. The inhibition was reversed by addition of bovine serum albumin, β-cyclodextrin or spermine.

     

Targeted Analysis of Lysosomal Directed Proteins and Their Sites of Mannose-6-phosphate Modification

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Highlights

• •Fe³⁺-IMAC enables co-enrichment of mannose-6-phosphate (M6P) modified glycopeptides.
• •Detailed characterization of intact M6P modified glycopeptides from HeLa and CHO cell lines.
• •EThcD results in a cleavage between 2 core GlcNAc residues enabling unambiguous glycoform identification.
• •Double knock out of the phosphatases Acp2/5 results in a 20-fold increase in abundance of M6P modified glycopeptides.