Phosphonomonoesterase activity in Metridium senile L.

• 1.1. An esterase which hydrolyzes 4-nitrophenyl(phenyl)phosphonic acid (4-NPPP) was purified from M. senile (sea anemone).
• 2.2. The enzyme showed no 5′-nucleotide phosphodiesterase activity with 5′-(4-nitrophenyl) TMP or phosphomonoesterase activity with 4-nitrophenylphosphate.
• 3.3. Addition of excess Zn²⁺ restored activity after inactivation by EDTA.
• 4.4. Thiol reagents and phenylmenthanesulfonylfluoride did not inactivate, whereas, dithiothreitol inactivated.
• 5.5. Aminoethylphosphonic acid (AEP) was a competitive inhibitor of 4-NPPP indicating possible activity with phosphonomonoesters of AEP.

     

Short-term osmotic responses of cells and tissues of the sea anemone,Condylactis gigantea

• 1.1. Cells of tentacles and body wall of the sea anemone Condylactis gigantea behaved as simple osmometers during 5hr exposure to 50, 67, 83, 100 and 125% sea-water.
• 2.2. All intracellular water appeared to be osmotically active.
• 3.3. Cell sodium, chloride and total osmolyte content remained invariable, with taurine decreasing and potassium increasing as sea-water concentration was reduced.
• 4.4. Tissues, as a whole, exhibited a pseudoregulatory response to changes in salinity as the large and osmotically inert extracellular space buffered volume changes to a considerable extent.

     

Time related changes in the free amino acid pool of the sea anemone,Bunodosoma cavernata,during salinity stress

• 1.1. The sea anemone, Bunodosoma cavernata, is a relatively eurybaline cnidarian tolerating salinities from 12 to 40%.
• 2.2. Taurine, glutamic acid and aspartic acid all showed some increases with increased salinity.
• 3.3. The amino acid showing the greatest accumulation under high salinity conditions was β-alanine which increased 28-fold from 1.5 to 41.9 μmol/g dry weight when salinity was raised from 26 to 40%.
• 4.4. When B. cavernata was subjected to increased salinity, β-alanine was rapidly accumulated and reached maximum levels within 4 days.
• 5.5. When salinity was dropped from 36 to 26%0, β-alanine concentrations dropped from 15 to 2 μmol/g dry weight in 2 days.

     

5-OH-Dopa and 5-S-cysteinyldopa: New biogenic amino acids in invertebrates

• 1.1. 5-OH-Dihydroxyphenylalanin (5-OH-Dopa) was found with the aid of high-pressure liquid chromatography (HPLC) and GC-MS to be present in four species of sea anemones.
• 2.2. 5-S-Cysteinyldopa was also found in the sea anemones as well as in the epidermis of two crustacean species. The results were obtained with HPLC and, in the case of the crustaceans, also verified spectro-fluorometrically.
• 3.3. Purified tyrosinae from the sea aneemone Metridium senile and the crayfish Pacifastacus leniusculus had a significant capacity for oxidizing Dopa to 5-S-cysteinyldopa and 2-S-cysteinyldopa in the presence of cystein. In the absence of cystein 5-OH-Dopa was formed only by the sea anemone.

     

Isolation of crustacean egg yolk lipoproteins by differential density gradient ultracentrifugation

• 1.1. Egg yolk lipoproteins from four species of Crustacea were isolated by differential density gradient ultracentrifugation.
• 2.2. Egg yolk proteins from freshwater prawn, striped stone crab and mitten crab consissted of high-density lipoprotein (HDL) and lipid-free protein, while low-density lipoprotein (LDL) was present in the egg yolk protein of sand crayfish as well as HDL and lipid-free protein.
• 3.3. HDL was a major component in the egg yolk proteins from four species of Crustacea. HDL was identical to egg yolk lipovitellin.
• 4.4. Both HDL and LDL possessed phospholipid as a major lipid.
• 5.5. HDL, but not LDL, contained carotenoids. The color of HDL from mitten crab showed a reddish purple and was distinct from other Crustacea whose color was orange. The reddish purple color was characterized by an absorption flexion at 600–650 nm.

     

A halogenated amino acid-containing sperm activating peptide and its related peptides isolated from the egg jelly of sea urchins,Tripneustes gratilla,Pseudoboletia maculata,Strongylocentrotus nudus,Echinometra mathaei and Heterocentrotus mammillatus

• 1.1. Twenty-eight peptides were isolated from the egg jelly of sea urchins, Tripneustes gratilla, Pseudoboletia maculata, Strongylocentrotus nudus, Echinometra mathaei (type A and B) and Heterocentrotus mammillatus and their amino acid sequences were determined.
• 2.2. Two of the peptides obtained from T. gratilla egg jelly possessed a bromophenylalanine (Br-Phe) residue in their sequences (Gly-(Br-Phe)-Asn-Leu-Asn-Gly-Gly-Gly-Val-Gly and Gly-(Br-Phe)-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly).
• 3.3. All of the peptides elevated cyclic GMP concentrations in the spermatozoa of the respective sea urchin and caused a shift in the apparent mol. wt of a major sperm protein of the respective sea urchin.
• 4.4. They stimulated respiration rates of the spermatozoa of Hemicentrotus pulcherrimus as well as their own species.
• 5.5. One-half maximal concentrations of the peptides for respiratory stimulation of H. pulcherrimus spermatozoa were between 10⁻¹¹ M and 10⁻⁹ M except a methionine-containing peptide which was about 10⁻⁷ M.

     

The carotenoids of eggs of wild and farmed cod (Gadus morhua)

• 1.1. Eggs of wild cod, and of farmed cod fed (a) a diet supplemented with astaxanthin and (b) a diet supplemented with both astaxanthin and canthaxanthin, were analysed with respect to carotenoids.
• 2.2. The total carotenoid contents in eggs were 0.7 ppm for wild cod and 0.5 ppm for farmed cod.
• 3.3. Cod, having white flesh, deposit ketocarotenoids in the eggs, preferably astaxanthin.
• 4.4. Canthaxanthin can replace astaxanthin in the eggs, but astaxanthin appears to be deposited preferentially when both carotenoids are present in the diet.
• 5.5. The isomer distribution of (3S, 3′S):(3R, 3′S, meso):(3R, 3′R) astaxanthin in the eggs reflected the isomer composition of the diet.
• 6.6. Echinenone, 4′-hydroxyechinenone, adonixanthin and zeaxanthin encountered in cod eggs may represent reductive metabolites of canthaxanthin and astaxanthin.

     

Comparative biochemical studies of carotenoids in sea-urchins—II. The more primitive sea-urchins belonging to the orders Cidaroida,Echinothurioida, Diadematoida and Arbacioida

• 1.1. The carotenoids of seven species of more primitive sea-urchins, [orders Cidaroida (I), Echinothurioida (II), Diadematoida (III), and Arbacioida (IV)] were investigated from the comparative biochemical point of view.
• 2.2. β,β-carotene and β-echinenone have been isolated as major carotenoids in (I) and (III, IV), respectively. In (II), β,β-carotene, β-echinenone, canthaxanthin and (3S,3′S)-astaxanthin were foundto be predominant carotenoids.
• 3.3. The carotenoid patterns of (I) which is the most primitive sea-urchin from the phylogenetic point of view, and of (II) which is direct developers with non-feeding larvae, were quite different from those of the other sea-urchins showing typical development with feeding larvae.

     

Metabolism of carotenoids in sea-urchin Pseudocentrotus depressus

• 1.1. Feeding experiments with β,β-carotene, canthaxanthin and astaxanthin on the sea urchin Pseudocentrotus depressus were investigated.
• 2.2. In the case of β,β-carotene group, β-carotene was accumulated, β-isocryptoxanthin appeared and β-echinenone increased 6.8 times as much as the control group. On the other hand, in canthaxanthin and astaxanthin groups, canthaxanthin and astaxanthin increased significantly, respectively. The metabolic products of these carotenoids could not be found.
• 3.3. It was concluded that β,β-carotene was bioconverted to β-echinenone via β-isocryptoxanthin in P. depressus and could not be oxidatively metabolized beyond β-echinenone.

     

Characteristics of carotenoid features in muscle and ovary from anadromous and river resident types of Alaskan dolly varden charr (Salvelinus malma malma)

• 1.1. The carotenoids in the muscles and eggs from two types of natural Alaskan dolly varden charr, anadromous and river resident types, were examined.
• 2.2. In the muscle from the anadromous charr, astaxanthin was the major component comprising more than 70% of the total, followed by idoxanthin and 4-keto-zeaxanthin.
• 3.3. The egg carotenoid features in the river resident charr were more complicated compared with those in the anadromous fish. A considerable proportion of unidentified carotenoids was found in the eggs from the river resident charr.
• 4.4. Idoxanthin was the main component along with considerable propotions of β-carotene tetrol and astaxanthin in the eggs from the anadromous charr, whereas zeaxanthin and lutein were detected besides idoxanthin and β-carotene tetrol in the eggs from the river resident fish.
• 5.5. The prey was considered to be responsible for the difference in the carotenoid features of the eggs from the anadromous and river resident charr.

     

Stiffness of starfish arm and involvement of catch connective tissue in the stiffness change

• 1.1. Bending tests were performed on intact arms of two starfishes of different orders to obtain stiffness of the arm.
• 2.2. Linckia laevigata without stimulation showed a wide variety of arm stiffness of 2.24–51.3 MPa (average: 8.0 MPa). Mechanical stimulation increased the stiffness by 2.5 times.
• 3.3. In Asterias forbesii, isolated arms were 23 times stiffer than intact ones. Anesthesia with 0.1% MS-222 or menthol-saturated sea water increased the stiffness by 7–170 times.
• 4.4. Ion dependence of stiffness suggests that the catch connective tissue was involved in the stiffness change.

     

Driving forces for the uphill transport of amino acids into epidermal brush border membrane vesicles of the sea anemone,Anemonia sulcata (Cnidaria,anthozoa)

• 1.1. The transport of amino acids into membrane vesicles prepared from epidermal tentacle tissue of the sea anemone, Anemonia sulcata, depends on an electrochemical potential difference caused, e.g. by sodium chloride gradients.
• 2.2. Potassium or choline chloride gradients energized the transport less effectively than sodium chloride gradients. Both Na⁺-ions and Cl⁻-ions were required for the amino acid transport.
• 3.3. The uphill transport of amino acids along the downhill movement of driver ions (sodium chloride gradient conditions) was characterized by an overshoot; under sodium chloride equilibrium conditions, however, an accumulation of amino acids within the vesicles could not be measured.
• 4.4. Potassium diffusion potentials in combination with valinomycin indicated that hyperpolarization (vesicle inside negative) and hypopolarization (vesicle inside positive) enhanced or depressed the accumulation of amino acids within the vesicles.
• 5.5. Being at the phylogenetic base of the Eumetazoa, cnidarians show characteristics for the transmembrane transport of amino acids comparable to those established for vertebrates.

     

Na+/H+ exchange in the kidneys of eels (Anguilla anguilla) adapted to sea water or to freshwater environments: Studies with brush border membrane vesicles

• 1.1. In brush border membrane vesicles isolated from eel kidneys, adapted either to sea water or freshwater environments, a Na⁺/H⁺ antiporter is present.
• 2.2. Using a calibration plot it is possible to evaluate the amount of protons that this antiporter can accumulate inside the vesicular space.
• 3.3. The activity of the antiporter seems to be affected by the salinity of the water; it is higher in animals adapted to seawater.
• 4.4. This adaptation seems to occur by a J_max regulation of the antiporter.

     

Microheterogeneity of late histones in larval stages of sea urchin development

• 1.1. The total histone complement of early plutei were compared with that of intermediate and late larvae of the sea urchin Tetrapygus niger.
• 2.2. Electrophoretic comparison indicates that there are quantitative and qualitative shifts of the five classes throughout late larval development.
• 3.3. The strong similarity in the amino acid composition of total histones isolated from early, intermediate and late plutei indicates that the observed electrophoretic heterogeneity is due to post-translational modifications.

     

Isolation of dna and rna from Streptococcus sobrinus OMZ176 using CsTFA gradients

• 1.1. A simple procedure for isolation of high molecular weight genomic DNA, and RNA, from Streptococcus sobrinus OMZ176 is described.
• 2.2. Cell cultures were grown aerobically for 10 hr.
• 3.3. Spheroplast formation and lysis was achieved by mutanolysin/lysozyme-dependent digestion of the cell wall, followed by N-lauroylsarcosinate-mediated lysis.
• 4.4. Nucleic acids were isolated directly from cell-lysates using cesium-trifluoroacetate (CsTFA) densitygradient centrifugation.
• 5.5. Three different centrifugation regimes were tested: self-generated density gradients in a fixed angle rotor; self-generated density-gradients in a swinging-bucket rotor; pre-formed density-gradients in a swinging-bucket rotor.
• 6.6. Genomic DNA isolated by the CsTFA-procedure was found to have higher purity as compared to genomic DNA isolated in a conventional CsCl gradient.
• 7.7. Isolated DNA was shown to be of a quality suitable for applications in molecular biology.

     

Purification and characterization of hatching enzyme of the pike (Esox lucius)

• 1.1. A choriolytic enzyme was isolated from the hatching medium of the pike, Esox lucius.
• 2.2. The enzyme is defined as hatching enzyme.
• 3.3. The molecular weight of the enzyme is 24,000.
• 4.4. The enzyme is a glycoprotein containing 2% carbohydrate.
• 5.5. Its isoelectric point is 6.5.
• 6.6. The pH optimum is around pH 8.
• 7.7. The enzyme molecule contains two disulfide bonds but no free cysteine.
• 8.8. Inhibitor studies and metal analysis show that the enzyme is a zinc-metalloprotease.

     

Characterization of histones from plutei larvae of the sea urchin Tetrapygus niger

• 1.1. Histones were isolated from plutei larvae of the sea urchin Tetrapygus niger and analysed electrophoretically. Individual histones were purified and their amino acid compositions were determined.
• 2.2. The electrophoretic analysis revealed that larval histones are microheterogeneous; H1 exhibits four subforms, the nucleosomal core histones H2A, H2B and H3 were resolved into three subforms each and H4 had two subforms.
• 3.3. The comparisons of the amino acid compositions of plutei larvae histones with data from the literature of homonimus late variants isolated from gastrulas of other sea urchin species, indicate that late histone variants are conserved proteins with a very slight degree of species specificity and with general features of classical histones.

     

Sea water drinking as a homeostatic response to dehydration in hatchling loggerhead turtles Caretta caretta

• 1.1. Hatching Caretta caretta may lose up to 12% of their initial hatched weight from water loss during emergence from the nest.
• 2.2. After subsequent osmotic and excretory water loss in sea water, hatchlings will drink sea water (166 μl 100 g⁻¹ hr⁻¹) and return to their initial weight within 10–15 days, without feeding.
• 3.3. There were no significant changes in plasma osmolarity or sodium levels over this period.
• 4.4. This osmoregulatory strategy is in marked contrast to that seen in the estuarine crocodile, Crocodylus porosus.

     

Comparative study of proteolytic enzymes in the digestive tracts of the european sea bass and hybrid striped bass reared in freshwater

• 1.1. A comparative study of the proteolytic activity in four different sections of the digestive tracts of the European sea bass (Dicentrarchus labrax) and hybrid striped bass (Morone chrysops × M. saxatilis) reared in freshwater revealed minor differences between these fish.
• 2.2. Tryptic activity plays a major role in the proteolytic process in both fish.
• 3.3. The activity of seven intestinal proteolytic enzymes was detected utilizing a combination of specific substrates and inhibitors.
• 4.4. High levels of proteolytic activity were detected in both the proximal and distal sections of the fish intestine at a high pH range (9–10).
• 5.5. In situ monitoring of pH levels revealed a lower pH level in the intestinal proximal section of hybrid striped bass compared with the distal section.
• 6.6. In contrast, higher pH levels were detected at the proximal compared with the distal sections of D. labrax intestine.

     

Adaptations of Crocodylus acutus and Alligator for life in saline water

• 1.1. American crocodiles (C. acutus) weighing less than 200 g are unable to grow when kept in 35 ppt sea water in the laboratory. Yet paradoxically there are some highly saline areas in south Florida where rapid growth occurs. It is possible that these conflicting observations can be reconciled by behavioral osmoregulation of young crocodiles.
• 2.2. Hatching occurs during the rainy season and small crocodiles may drink from the brackish “lens” available during and after rainfall.
• 3.3. Using a weekly regime of alternating exposure to 35 ppt (6 days) and 4 ppt (12–24 hr), it has been demonstrated that growth of small crocodiles occurs. Feeding takes place primarily when brackish water is available. Salinities as high as 18 ppt were drunk when crocodiles were dehydrated by 15–20% of initial mass.
• 4.4. C. acutus and Alligator have a rather low rate of water efflux in sea water (0.2ml/100g-hr).
• 5.5. Sodium influx in sea water of C. acutus is low, but higher than efflux. Thus there is no evidence yet for a significant role of the lingual salt glands in sodium excretion.
• 6.6. The major adaptations to saline water of hatchling C. acutus are a low intake of sodium, an ability to selectively drink water of lower salinities, and to grow very rapidly (within 3–4 months) to a size much more tolerant of immersion in 35 ppt sea water.