Sterol composition of two actiniaria

• 1.1. The sterol composition of Condylactis aurantiaca and Cereus pedunculatus (phylum Coelenterata, class Anthozoa, order Actiniaria) was investigated by silver nitrate-silica gel column chromatography, combined gas chromatography-mass spectrometry and NMR.
• 2.2. Sea anemones contained Δ³-sterols accompanied by small amounts of Δ^5.7-sterols and ring saturated sterols.
• 3.3. Sterols with 27 carbon atoms are predominant and cholesterol is the principal sterol, followed by 24-methylenecholesterol.
• 4.4. A 4-methyl ring saturated sterol, identified as 4,24-dimethyl-5α-cholest-24(28)-en-3β-ol, occurs in small amount in Actiniaria, accompanied by the corresponding 4-demethylstanol.

     

Steroid metabolites of the far eastern Holothurian Stichopus japonicus Selenka

• 1.1. Sulphated and etherified sterols were isolated from the far eastern holothurian Stichopus japonicus S. The sterol composition of both fractions was determined using gas-liquid chromatography and mass-spectroscopic methods. The structures of individual sterols were proved on the basis of mass-spectrometry and ¹H-NMR-spectroscopy data.
• 2.2. The structures of 29 sterols were established.
• 3.3. Sterols (22E, 24R)-23,24-dimethyl-5α-cholest-22-en-3β-ol, 23,24-dimethyl-cholesta-5,22-dien-3β-ol, 24-methyl-cholesta-5,24(28)-dien-3β-ol, (24Z)-24-ethyl-cholesta-5,24(28)-dien-3β-ol, 24-nor-cholesta-5,22-dien-3β-ol, 24-ethyl-cholesta-5,25-dien-3β-ol were described for holothurians for the first time.
• 4.4. Δ⁵-sterols were shown to be the main components of the sulphated alcohol fractions (67.61%), while the saturated and Δ⁷-sterols were there in less quantities (14.72 and 9.52%, respectively).
• 5.5. The etherified sterols were represented, mainly, by saturated and Δ⁷-sterols (37.82% and 33.95%, respectively). Δ⁵-sterols were 19%.
• 6.6. The sensitivity of liposomal membranes, containing steroid metabolites of the holothurian St. japonicus (Δ⁷-, sulphated and glycosilated sterols) to the action of endotoxin-stichoposide A, was studied.

     

Δ5,7Sterols from the sponges Ircinia pipetta and Dysidea avara identification of cholesta-5,7,24-trien-3B-OL

• 1.1. Δ^5,7-sterols have been isolated as pure compounds from the marine sponges Ircinia pipetta (Dictyoceratida:Thorectidae) and Dysidea avara (Dictyoceratida:Dysideidae) by reverse phase HPLC and analyzed by GLC, u.v., mass spectrometry and ¹H-NMR.
• 2.2. Ircinia pipetta and D. avara have rather similar sterol compositions and contain predominantly Δ^5,7-sterols, accompaned by Δ⁵-sterols. Ergosterol, cholesta-5,7-dien-3β-ol and 24-ethylcholesta-5,7-dien-3β-ol are the major sterols in I. pipetta, while D. avara contains in addition to these three sterols, (24Z)-24-ethylcholesta-5,7,24(28)-trien-3β-ol as the fourth major sterol.
• 3.3. Cholesta-5,7,24-trien-3β-ol which previously was not isolated from a marine organism is also present.

     

The sterol constituents and Δ5,7-sterol content of some bivalve molluscs

• 1.1. The bivalve molluscs Cerastoderma edula, Chlamys opercularis, Ensis soliqua, Modiolus modiolus, Mya arenaria, Mytilus edulis and Pecten maximus contained mixtures of C₂₆-, C₂₇-, C₂₈- and C₂₉-sterols. Cholesterol, 24-methylcholesta-5,22-dien-3β-ol and 24-methylene-cholesterol were the major sterols.
• 2.2. The sterols of Cerastoderma edula, Mya arenaria and Mytilus edulis contained 6–16% of cholesta-5,24-dien-3β-ol.
• 3.3. All the molluscs contained Δ^5,7-sterols in amounts ranging from 2 to 21% of the total sterols.
• 4.4. Cholesta-5,7-dien-3β-ol and 24-methylcholesta-5,7,22-trien-3β-ol were identified in Mytilus edulis. 25-Norcholesta-5,7,22-trien-3β-ol was detected in Modiolus modiolus.

     

Sterol composition of the “living fossil” crinoid Gymnocrinus richeri

• 1.1. The composition of sterol mixture from the “living fossil” crinoid Gymnocrinus richeri collected off Nouméa (New Caledonia) was investigated.
• 2.2. The free 3β-OH sterol mixture was found to contain 14 components, Δ⁵ and ring saturated stanols, identified by GC-MS.
• 3.3. Cholest-4-en-3-one, cholesta-1, 4-dien-3-one (this latter firstly isolated from a marine source), 5α-8α-epidioxy sterols, and 5α-ergosta-7,22-diene-3β,5,6β-triol were also present, their characterization being accomplished by EI-MS and ¹H-NMR. The methanol extract also contained sterol sulphates, which were identified by GC-MS after solvolysis to remove the sulphate group.

     

Comparative study of the sterol composition of the digestive gland and the gonad of Sepia officinalis L. (mollusca,cephalopoda)

• 1.1. The sterol composition of the digestive gland and the gonad of Sepia officinalis L. was investigated by GC and GC-MS.
• 2.2. The same sterols were recognized in both organs, cholesterol being the major component of the sterol mixtures. However, quantitative differences appeared between the sterol composition of the digestive gland and the gonad.
• 3.3. The sterol mixtures of the digestive gland and the gonad of immature and mature females and males of various origins were compared. Quantitative changes in the sterol composition of the gonad were related to sexual maturity whereas the sterol composition of the digestive gland appeared linked to the diet.

     

Sterol composition of marine bivalves from the genus Macoma

• 1.1. Four members of the genus Macoma: M. Balthica, M. irus, M.;incongrua; and M. contabulata from the Japan Sea were investigated for their sterol composition.
• 2.2. Cholest-5-en-3β-ol was the most abundant sterol in all investigated animals; the other major sterols were common constituents of bivalves.
• 3.3. The observed similarity in sterol composition of the studied clams seems to be an indication of greater influence of ecological than genetic factors on sterol composition.

     

Sterol composition of three marine sponge species from the genus Cinachyrella

• 1.1. The hitherto undescribed sterol compositions of three marine sponge species belonging to the genus Cinachyrella are reported: C. alloclada and C. kükenthali from the Senegalese coast, at two different depths, and C. aff. schulzei from the lagoon of Nouméa, New Caledonia.
• 2.2. Fourteen free sterols have been identified by GC and GC/MS studies, including the 23,24ξ-dimethylcholesta-5,22-dien-3β-ol (10) and the rare 24-norcholesta-5,22-dien-3β-ol (1).
• 3.3. The first compound (10) is reported for the second time in a marine sponge and it was found only in Senegalese sponges collected in shallow waters.
• 4.4. Sterol (10) has been isolated by HPLC and identified by NMR techniques.
• 5.5. Significant amounts of cholest-7-en-3β-ol (7) were also found in the Senegalese sponge species.
• 6.6. Apart from these two compounds, the three sponge sterol compositions are found to be very similar.

     

The influence of stigmasterol concentration on the growth and lipid composition of wild-type and barium a strains of Paramecium tetraurelia

• 1.1. The ciliary membrane lipid composition and electrophysiology of the behavioral mutant of Paramecium tetraurelia, barium A (d4–592), were previously shown to differ from those of wild-type 51S when both strains were grown in low sterol medium.
• 2.2. In this study, the phospholipid fatty acid composition of the two strains was shown to differ regardless of the level of sterol supplementation or culture age (growth phase).
• 3.3. The ratio of linoleic to γ-linolenic acid, 18:2(9,12)/18:3(6,9,12), was consistently higher in baA compared to wild-type phospholipids, largely because of a dramatic shift in the ratio of these two fatty acids esterified at the 2 position of 1-alkyl-2-acyl-sn-glycero-3-phosphorylcholine (GPC).
• 4.4. These data support the hypothesis that a specific Δ6 fatty-acyl desaturase, which directly desaturates phospholipid and shows a preference for GPC as its substrate, is impaired as a result of the barium A mutation.

     

Thermal properties for digestive enzymes of a sub-antarctic beetle,hydromedion sparsutum (coleoptera,perimylopidae) compared to those in two thermophilic insects

• 1.1. Proteolytic, lipolytic, amylolytic and cellulolytic activities were studied in adults of the phytophagous beetle, Hydromedion sparsutum, indigenous to the sub-Antarctic island of South Georgia.
• 2.2. Gastric enzyme activities were measured at experimental temperatures of 5–40°C and results were compared with those obtained from two thermophilic insects, Gryllus bimaculatus and Tenebrio molitor.
• 3.3. Protease and lipase activities in Hydromedion were 10–15 times lower than in Gryllus and Tenebrio.
• 4.4. In the temperature range of 5–15°C, α-amylase activity from Hydromedion was only slightly lower than that from Gryllus.
• 5.5. Hydromedion gut homogenates exhibited a distinct cellulolytic activity, even at a low temperature of 5°C.
• 6.6. Cellulolytic activity in the digestive tract of Hydromedion was confirmed by the evolution of ¹⁴CO₂ after consumption of labelled cellulose.
• 7.7. The thermal properties of digestive enzymes agree well with the role of Hydromedion as primary decomposer in its ecosystem.

     

The regulation of glucose 6-phosphate dehydrogenase from Dicentrarchus labrax (bass) liver

• 1.1. Kinetic constant values of the reaction catalyzed by bass liver glucose 6-phosphate dehydrogenase show to be modified between 10 and 40°C.
• 2.2. The Arrhenius plot between 10 and 50°C shows two slopes with different activation energies.
• 3.3. These results suggest a regulation of this enzyme by environmental temperature.
• 4.4. Kinetics of ATP inhibition were examined between pH 6.2 and 7.8: patterns and K_i values obtained are affected by the pH variation.
• 5.5. NADH is an effective inhibitor of bass glucose 6-phosphate dehydrogenase but this enzyme does not show NAD-linked activity.
• 6.6. Kinetics of pyridoxal 5′-phosphate inhibition have indicated the presence of a lysine in the catalytic site for NADP⁺.

     

An alkaline phosphatase from Thermus sp strain Rt41A

• 1.1. A thermostable orthophosphoric monoester phosphohydrolase (EC 3.1.3.1) from Thermus sp strain Rt41A has been purified 400-fold to give a specific activity of 25 U/mg at 60°C in IM diethanolamine (pH 11.1).
• 2.2. The enzyme has a M_r of 160,000 and is trimeric.
• 3.3. The half-life of the enzyme is 5 min at 85°C.
• 4.4. The enzyme has a wide specificity for a number of phosphate monoesters.
• 5.5. The H_m of the enzyme is pH dependent, so the pH optimum of the enzyme is affected by the substrate concentration.
• 6.6. The enzyme is inhibited 50% by 20 mM Ca²⁺ or Mg²⁺.
• 7.7. The K_i for phosphate, EDTA-di sodium salt and arsenate (in 1 M diethanolamine, pH 11.1) is approx 1.2, 1.6 and 4mM respectively.
• 8.8. Urea (200 mM) is not inhibitory.

     

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.

     

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.

     

Alkaline phosphatase from Basidiobolus haptosporosus: Comparison with alkaline phosphatases from rainbow lizard and man

• 1.1. DEAE-cellulose chromatography of mycelial alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) from Basidiobolus haptosporosus, produced three iso-enzymes “A”, “B” and “C”.
• 2.2. Fraction “A” was further characterized and showed maximum activity at pH 10 in 0.1 M sodium carbonate-bicarbonate buffer.
• 3.3. The enzyme was stimulated by Mg²⁺, Co²⁺ and Mn²⁺ and inactivated by Zn²⁺, Cu²⁺, EDTA, citrate and tartrate.
• 4.4. Phosphate ions inhibited it competitively, phenylalanine uncompetitively and urea noncompetitively.
• 5.5. It was heat stable for 60 min at 37°C but labile above 55°C.
• 6.6. Its K_m with p-nitrophenylphosphate was 0.5 mM; its estimated molecular weight was 160,000.
• 7.7. The results are compared with the properties of alkaline phosphatases from the rainbow lizard and man and discussed in terms of a triadic association between the fungus, the lizard and man.

     

Behavioural thermoregulation and critical thermal limits of Macrobrachium acanthurus (Wiegman)

• (1)The preferred temperatures of Macrobrachium acanthurus were determined for prawns acclimated to 20°C, 23°C, 26°C, 29°C and 32°C, and the final preferendum estimate was (29.5°C).
• (2)The critical thermal minima (CTMin) and maxima (CTMax) were 11.0°C, 12.1°C, 13.0°C and 14.8°C, and 34.2°C, 35.0°C, 36.1°C and 39.8°C, respectively.
• (3)The zone of thermal tolerance assessed using the CTMin and CTMax boundaries was 644°C².
• (4)The acclimation response ratio was between 0.33 and 0.62.
• (5)To cultivate this species in the southeastern region of México it should be done in not <15°C (CTMin) during the winter and below 38°C in summer (CTMax).

     

In vitro substrate utilization for lipid synthesis in liver explants from hyperthyroid chickens

• 1.1. Indian River male broiler chickens growing from 7 to 28 days of age were fed diets containing 12, 18, 24 and 30% protein + 0 or 1 mg triiodothyronine (T₃)/kg of diet to study energetic costs of lipogenesis and the use of various substrates for in vitro lipogenesis.
• 2.2. De novo lipid and CO₂ production were determined in the presence of [1-¹⁴C]pyruvate, [2-¹⁴q]pyruvate, [3-¹⁴C]pyruvate, [2-¹⁴C]acetate and [U-¹⁴C]alanine.
• 3.3. Oxygen consumption was determined in mitochondrial preparations to estimate the energetic costs in expiants synthesizing lipid.
• 4.4. Radiolabeled CO₂ derived from [1-¹⁴C]pyruvate was used as an estimate of coenzyme A availability in liver expiants. Lipids derived from [2-¹⁴C]pyruvate, [2-¹⁴C]acetate and [U-¹⁴C]alanine estimate relative substrate efficiency.
• 5.5. Labeled CO₂ production from [1-¹⁴C]pyruvate was greatest in that group fed a 12% protein diet and least in the group fed a 30% protein diet.
• 6.6. In addition, T₃ increased CO₂ production from [1-¹⁴C]pyruvate.
• 7.7. The production of ¹⁴CO₂ from the second carbon of pyruvate or acetate was increased by T₃.
• 8.8. The low-protein diet (12% protein) increased (P <0.05) lipogenesis.
• 9.9. Adding T₃ to the diets decreased carbon flux into lipid from all substrates, but increased CO₂ production from all substrates without changing stage 3 and 4 respiration rates in mitochondrial preparations.
• 10.10. These observations imply that coenzyme A availability may have regulated de novo lipogenesis in the present study.
• 11.11. It was also concluded that previously noted effects of T₃ on intermediary metabolism may involve metabolic pathways that do not involve changes in mitochondrial function.

     

Light-induced transient increase of the activity of topoisomerase I in plasmodia of Physarum polycephalum

• 1.1. Activity of topoisomerase I and incorporation of [³H]uridine and [¹⁴C]thymidine were monitored during light-induced sporulation of the slime mold Physarum polycephalun.
• 2.2. A 4-fold transient increase of topoisomerase I activity but not of [³H]uridine or [¹⁴C]thymidine incorporation was observed after 42 hr of illumination with 6 hr impulses.
• 3.3. The activity of topoisomerase I did not increase in the absence of light impulses. However, ca 5-fold increase of the activity was observed in dark when 100 μ M dibutyryl-cAMP was administered 12 hr before harvesting of plasmodia.
• 4.4. Fluorodeoxyuridine and cycloheximide administered 36 hr after starting of the illumination cancelled the increase of the activity of topoisomerase I.
• 5.5. After 7 days of the illumination, when fruiting bodies appeared, the activity of topoisomerase I dropped to about 15% of the initial value.

     

Sterols from black sea invertebrates—I. Sterols from Scyphozoa and Anthozoa (Coelenterata)

• 1.1. Rhizostoma pulmo, Aurelia aurita and Actinia equina, the most widespread representatives of Coelenterata in Black Sea have been analysed and the occurrence of 20 sterols has been found.
• 2.2. Dinosterol and demethyldinosterol as well as a number of short side chain sterols have been found in Scyphozoa for the first time.
• 3.3. The occurrence of coprostanol in marine invertebrates has been shown for the first time.
• 4.4. Five groups of sterol esters were found, containing fatty acids with different polarity.

     

Steroids from aquatic organisms—XII. Sterols from the antarctic sponge Homaxinella balfourensis (Ridley and Dendy)

• 1.1. The sterol composition of the sponge Homaxinella balfourensis (Ridley and Dendy) has been analysed and seven components detected.
• 2.2. These were separated by argentic column chromatography and studied by gas chromatography-mass spectrometry and by proton magnetic resonance spectroscopy.
• 3.3. It was established that the components were C₂₇, C₂₈ and C₂₉ fully saturated or side chain unsaturated stanols and colest-5-en-3β-ol as traces.