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).

     

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).

     

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.

     

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.

     

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.

     

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.

     

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.

     

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.

     

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.

     

Unfolding and release of heme from human hemoglobins A,S and F

• 1.1. Analysis of the Soret spectra of hemoglobins A, S and F has been used to determine the extent of heme exposure and release from these hemoglobins in the presence of several solvent perturbants.
• 2.2. Oxyhemoglobin S unfolding in the presence of either urea or propyl urea resulted in greater heme exposure and release than either oxyhemoglobins A or F.
• 3.3. Methemoglobin formation resulted in lower denaturation midpoints for each hemoglobin compared to the reduced oxyhemoglobin state; methemoglobin F had the lowest denaturation midpoint under isothermal denaturing conditions.
• 4.4. Rate of heme exposure was greater for oxyhemoglobin S than oxyhemoglobin A in the presence of 200 μM the anionic detergent sodium dodecyl sulfate.
• 5.5. Evidence for increased levels of heme release in hemoglobin S may be related to the greater tendency of sickled red cell membranes to undergo lipid oxidation.

     

The inhibitory effect of tetramethylethylene diamine on water soluble and membrane bound acetylcholinesterase activity

• 1.1. The inhibitory effect of N,N,N′,N′-tetramethylethylene diamine (TEMED) on water soluble (WSAChE) and membrane bound (MBAChE) acetylcholinesterase was investigated.
• 2.2. TEMED (0.5–4.0 mM) reversibly inhibited WSAChE activity (18–62%) and MBAChE (20–61%) in a concentration dependent manner.
• 3.3. The IC₅₀ being about 2.8 mM for WSAChE and 2.6 mM for MBAChE.
• 4.4. Lineweaver-Burk plots indicated that the nature of inhibition is noncompetitive for both water soluble and membrane bound acetylcholinesterase, with K_m values 68 μM and 123 μM respectively.
• 5.5. An Arrhenius plot showed that the transition temperature (TT) is unaffected in the presence of TEMED.
• 6.6. The activation energy was increased below and above TT in the case of WSAChE only.
• 7.7. On the basis of this behaviour of TEMED with AChE. it can be proposed that it can be used as an eluting agent for the bounded AChE to affinity ligand and may have beneficial action on the reactivatability of irreversibly-inhibited AChE due to its structure.
• 8.8. Moreover there is a possibility that it can be used as a therapeutic agent for the treatment of Alzheimer's disease, myasthenia gravia and glaucoma like some other inhibitors of AChE.

     

Effects of feeding,starvation, and refeeding on the fatty acid composition of channel catfish,ICtalurus punctatus,tissues

• 1.1. The effects of feeding, food deprivation (14 and 28 days) and refeeding (starved 14 then fed 14 days) on the fatty acid composition of white muscle, liver and brain of pond-raised channel catfish (Ictalurus punctatus) were investigated.
• 2.2. Levels of n-3 fatty acids were significantly higher (P < 0.05) in white muscle of fish starved 28 days (10.7%) than in fish fed throughout the study (8.0%), due primarily to an increase in 22:6(n-3) docosahexaenoic acid or DHA.
• 3.3. Significantly higher levels of 20:5(n-3) (eicosapentaenoic acid or EPA) were found in livers offish starved 28 days (P < 0.05) compared to fish fed throughout the study.
• 4.4. Results suggest that the fatty acid compositions of channel catfish white muscle and liver are subject to only limited perturbation during periods of starvation and refeeding and that the brain is extremely well protected.

     

Non-transferrin-bound-iron in serum and low-molecular-weight-iron in the liver of dietary iron-loaded rats

• 1.1. The feeding of 0.5% (3,5,5-trimethylhexanoyl)ferrocene (TMH-ferrocene) in rats resulted in a severe and progressive liver siderosis (total liver iron, 30 mg/g liver wet weight, after 30 weeks).
• 2.2. High concentrations of an iron-rich ferritin (up to 250 mg/l) were detected in serum of heavily iron-loaded rats forming a large fraction of non-transferrin-bound-iron (5000 μg/dl in maximum).
• 3.3. Ferritin and not haemosiderin was the major iron storage protein in the liver.
• 4.4. The total liver iron concentration (from 0.4 to > 30 mg Fe/g wet wt) but not the cytosolic low-molecular-weight-iron fraction (from 0.5 to 2.5 μM) was extremely increased during iron-loading.

     

Differential responses in cyclic GMP formation induced by excitatory amino acids (EAA) and sodium nitroprusside (SNP) in various regions of the brain and of rats of varied age

• 1.1. Sodium nitroprusside (SNP, 100 μM) caused a rapid and great increase of formation of cGMP in rat cerebellar slices. This effect was not blocked by l-nmma (a NO synthetase inhibitor) or antagonists of the NMDA receptor complex (e.g. AP₅ or MK 801).
• 2.2. Similarly, NMDA (100 μM) and glutamate (I mM) caused a rapid but less significant increase of cGMP formation. This increase was blocked by NMDA receptor complex blockers (e.g. AP₅, MK801 and kynurenate), and l-NMMA and l-nitroarginine.
• 3.3. In rats aged 12 days, both NMDA and kainate (at 100 μM) caused significantly increased levels of cGMP in the cerebellum, pons and medulla areas, whereas no significant alterations were found in the cerebral cortex, hippocampus or midbrain areas.
• 4.4. NMDA (100 μM) and SNP (300μM) induced greater increases of cGMP in cerebellar slices in young (aged 13 days) animals than older ones of either sex. This effect decreased greatly after 35 days of age. In adult (2 months) animals the effect of NMDA had virtually disappeared whereas SNP was barely significantly present.
• 5.5. Our results suggest that brain region and age, but not sex, affected formation of cGMP induced by excitatory amino acids (EAA) and SNP. Furthermore, endogenous NO production is required by EAA, but not by SNP, in the formation of cGMP.