Transport of cholesterol in the hemolymph of the mollusc Diplodon delodontus

• 1.1. The dietary and inter-organ cholesterol transport in the hemolymph of the bivalve mollusc Diplodon delodontus, was studied. Plasma and hemocytes were obtained after feeding labeled cholesterol to animals or injecting it into the posterior adductor muscle.
• 2.2. In both cases, cholesterol was incorporated either into plasma or hematic cells.
• 3.3. Two plasmatic fractions differing in their hydrated densities were recognized as cholesterol carriers and were isolated. They have characteristics of high density (HDL) and very high density (VHDL) lipoproteins, respectively.
• 4.4. The major lipids in the different classes of lipoproteins were free sterols in HDL and phospholipids in VHDL.
• 5.5. Neither low nor very low density lipoprotein transporting cholesterol was detected.

     

Sex-specific gene expression in distinct tissues of the shrimp Penaeus vannamei

• 1.1. Soluble proteins extracted from male and female Penaeus vannamei tissues such as eyes, eyestalks, brain, nerve cord, hemolymph, heart, muscle, hepatopancreas, hepatopancreas membrane and cuticular epidermis were analyzed and compared by high-resolution mini-two-dimensional polyacrylamide gel electrophoresis (mini-2D-PAGE).
• 2.2. In each shrimp tissue a large number of discrete polypeptides was observed.
• 3.3. The polypeptide patterns from the same tissue of female and male shrimp were mostly similar but both qualitative and quantitative differences were noted, suggesting the presence of sex-specific gene products in various shrimp tissues.
• 4.4. Future applications of these results are discussed.

     

The occurrence and distribution of haemoglobin in the entosymbiotic rhabdocoel Paravortex scrobiculariae (Graff) (Platyhelminthes: Turbellaria)

• 1.1. The occurrence of haemoglobin in the entosymbiotic turbellarian Paravortex scobiculariae has been demonstrated.
• 2.2. The haemoglobin occurs intracellularly in the mesenchyme, around the extrinsic musculature of the pharynx, and in the brain.
• 3.3. Haemoglobin could not be detected in Scrobicularia plana, the bivalve molluscan host of P. scrobiculariae, nor in the other British species of Paravortex, P. cardii, which is entosymbiotic in the bivalve Cerastoderma edule.
• 4.4. It is suggested that the haemoglobin of P. scrobiculariae is of functional significance when the entosymbiont is feeding at low tide in conditions of reduced oxygen tension.

     

Cellulose acetate electrophoresis of soluble lens proteins of different species of mammals; the use of discontinuous buffer systems

• 1.1. Cellulose acetate electrophoresis using a Tris-veronal discontinuous buffer system, has been performed with soluble lens proteins of different species of mammals and compared with two conventional continuous buffers.
• 2.2. In the four species investigated (mouse, Mus musculus; rat, Rattus norvegicus; bull, Bos taurus; ovine, Ovis aries) a higher migration rate and an improved fractionation were obtained with the discontinuous buffer.
• 3.3. The advantages and application to the lens protein research is indicated.

     

Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis (L.)

• 1.1. Anaerobic energy metabolism was investigated in different organs of Mytilus edulis and the whole animal.
• 2.2. Succinate accumulates to high levels in most organs but remains low in the hemolymph.
• 3.3. After 16 hours propionate accumulation is observed in all organs. Experimental evidence is not sufficient yet to point out organs that produce more propionate than others.
• 4.4. Acetate is a minor end product.
• 5.5. Acetate and propionate are found in the hemolymph in amounts equal to those in the organs.
• 6.6. Animals incubated in oxygen-free seawater accumulate more end products than animals exposed to air, in the form of volatile fatty acids that are excreted into the incubation water.
• 7.7. Alanine and glutamine increase in the posterior adductor muscle. Aspartate decreases in the total animal, posterior adductor muscle and gills, while in the hemolymph decrease in alanine, asparagine, serine, threonine and proline are observed.

     

Preparation and properties of mitochondria from tissues of the sea mussel Mytilus edulis L

• 1.1. A method is described for the preparation of coupled mitochondria from hepatopancreas, mantle and adductor muscle of the sea mussle, Mytilus edulis.
• 2.2. NADH is a non-penetrant, whereas succinate, glutamate and malate plus pyruvate have a clear stimulatory effect on respiration. Proline does not stimulate respiration of sea mussel mitochondria.
• 3.3. No P/0 ratios could be calculated as after the addition of ADP the mitochondria remain in the active state (state III), even after enough oxygen is consumed for complete phosphorylation. The reason for this observation is discussed.
• 4.4. Mitochondrial densities based on cytochrome b levels of two tissues of the mussel are compared to rat liver. Within the mussel the mantle contains twice the amount of cytochrome of adductor muscle.

     

The relation of d-alanine and alanine racemase activity in molluscs

• 1.1. Eighteen molluscan species were examined for the presence of d-alanine and alanine racemase activity to probe the probable relation between them.
• 2.2. Two bivalve species had high concentration of d-alanine and l-alanine (1:1) and showed high activities of alanine racemase. In these species, the occurrence of d-alanine could be explained by the action of alanine racemase.
• 3.3. In other species, the levels of d-alanine and enzyme activity were low, and the occurrence of d-alanine did not correspond with the presence of alanine racemase activity.
• 4.4. The mechanism of the occurrence of d-alanine in molluscan tissues seems to vary from species to species and seems not to be associated with the phylogenic situation or habitats of the respective species.

     

Allozyme variation in a wild african elephant (Loxodonta africana) population from the Kruger National Park,South Africa

• 1.1. Blood, liver, heart, testis, skin, eye, muscle and kidney samples were obtained from elephants (Loxodonta africana) in the Kruger National Park during a culling programme in April 1992.
• 2.2. Gene products of 25 protein coding loci in L. africana were examined by horizontal starch-gel electrophoresis.
• 3.3. Eighteen protein coding loci (72%) displayed monomorphic gel banding patterns whereas only seven (28%) displayed polymorphic gel banding patterns.
• 4.4. Average heterozygosity values for adults, youngsters and the total population are respectively 0.058, 0.024 and 0.047.
• 5.5. Relative gene diversities within and between populations are 84% and 16% respectively.
• 6.6. Two population simulation programmes were utilized to predict the duration of the current variability present in this species, based on current genetic variation and gene transfer from one generation to the next.

     

Detection of 180 kDa proteins in electroplax sodium channel preparations

• 1.1. Co-isolating proteins (M_r 170,000–220,000) from sodium channel preparations made from the electric organ of the electric eel (Electrophorus electricus) were detected on Western blots using monoclonal a antibodies.
• 2.2. Similar protein patterns were seen on immunoblots containing immunoprecipitated protein from eel muscle and brain tissues but not heart.
• 3.3. These co-isolating proteins could be separated from the mature TTX-sensitive channel protein (M_r 280,000) using a lentil lectin-Sepharose column.
• 4.4. The 180 kDa proteins do not appear to be channel-related and can be detected as contaminants in electroplax sodium channel preparations using the monoclonal antibodies described here.

     

Isolation and characterization of lipoamide dehydrogenase from mackerel dark muscle

• 1.1. Lipoamide dehydrogenase was purified 1500-fold from mackerel dark muscle.
• 2.2. The enzyme was homogeneous as judged by acrylamide gel electrophoresis in the presence and absence of SDS.
• 3.3. Molecular weights of 102,000 and 55,000 were estimated for the native and denatured enzyme, respectively.
• 4.4. Optimal activity for the enzyme was obtained at around pH 5.7 and enhanced with citri acid.
• 5.5. Loss of activity was less than 5% by incubating the enzyme at 70°C for 20 min.
• 6.6. An apparent K_m of 3.1 × 10⁻³ M was obtained for dl-lipoic acid and 1.5 × 10⁻⁵ M for NADH.
• 7.7. The properties of lipoamide dehydrogenase from mackerel dark muscle observed in this investigation were very similar to those reported for the enzyme from other sources.

     

Subcellular distribution of metals within the kidney of the bivalve Mercenaria mercenaria (L.)

• 1.1. The subcellular distribution of nine transition metals (plus four additional elements) was measured in the kidney tissue of the quahog, Mercenaria mercenaria.
• 2.2. Elemental analyses of the subcellular fractions indicated three main patterns of metal distribution within kidney cells.
• 3.3. Barium, iron, manganese and lead were associated primarily with kidney granules.
• 4.4. Cadmium, copper, potassium and magnesium were found mainly in the cytosolic fraction.
• 5.5. Calcium, phosphorus and zinc were found in all isolated fractions, probably reflecting the important roles that these elements play in bivalve metabolism.
• 6.6. The organelle composition of the isolated subcellular fractions was determined using marker enzyme assays and microscopic techniques.

     

Purification and some properties of isocitrate dehydrogenase of a blowfly Aldrichina grahami

• 1.1. NADH-dependent isocitrate dehydrogenase has been purified 110-fold from the crude extract of the flight muscle mitochondria of Aldrichina grahami.
• 2.2. The purification procedure involved Triton X-100 treatment of isolated mitochondria, column chromatography on DEAE-cellulose, Affi-gel blue, and P-cellulose.
• 3.3. The purified enzyme was homogeneous by criteria of the polyacrylamide gel electrophoresis.
• 4.4. The enzyme of the blowfly contains more acidic amino acids and less hydrophobic amino acids than that of pig heart.
• 5.5. The molecular weight was determined to be 330,000 daltons. The subunit construction differs from ghat of mammalian isocitrate dehydrogenase.

     

Purification and partial characterization of an AMP deaminase from the marine invertebrate Palaemon serratus

• 1.1. AMP deaminase from Palaemon serratus tail muscle was partially purified by chromatography on cellulose phosphate.
• 2.2. Muscle homogenates expressed very low enzyme activities and the presence of ATP was necessary to detect AMP deaminase. The specific activity and substrate affinity of the purified enzyme were also very low.
• 3.3. The purified prawn muscle AMP deaminase was contaminated by contractile proteins, one of the major contaminants being actin.
• 4.4. The enzyme displayed a very high affinity for actomyosin which was only partially abolished by pyrophosphate.

     

Electrophoretic characterization of a hybrid between Eretmochelys imbricata and Caretta caretta (Cheloniidae)

• 1.1. An intermediate morphotype between Eretmochelys imbricata and Caretta caretta was studied in Praia do Forte, Bahia, Brazil.
• 2.2. Three enzymatic systems were successfully analyzed: SOD, lactate dehydrogenase (LDH) and esterase (EST). Isoelectric focusing of total soluble proteins of muscle and transferrin were shown.
• 3.3. Esterase exhibited nine phenotype patterns, seven in C. caretta and one in the others morphotypes. SOD phenotypes were identical in the three morphotypes. Lactate dehydrogenase and transferrins were characteristic for each species.
• 4.4. Jaccard's measure of similarity was calculated and a phenogram with the three morphotypes were constructed using isoelectric focusing of total soluble proteins.

     

Age-dependent proteins in eyes of annual killifishes Pterolebias longipinnis detected by 2-dimensional electrophoresis

• 1.1. Eye proteins of Pterolebias longipinnis have been analyzed by 2-dimensional isoelectric focusing SDS-polyacrylamide gel electrophoresis during aging from adolescence until normal death.
• 2.2. The protein pattern on the gels changed gradually with progressing age.
• 3.3. In senescent eyes, three protein spots appeared for a time and 36 disappeared from the pattern.
• 4.4. The isoelectric points of the proteins in the presence of urea and the molecular weights in an unreducing buffer are presented.

     

Osmotic changes in an estuarine bivalve,Modiolus fluviatilis

• 1.1.Responses to different salinities monitored by opening and closing of the shell valves were observed in Modiolus fluviatilis.
• 2.2.The osmotic pressure, sodium and chloride ion concentrations were measured in the haemocoelic fluid of Modiolus fluviatilis under similar conditions.
• 3.3.Free amino acids (measured as ninhydrin-positive substances) were determined in the muscle tissue of Modiolus.
• 4.4.It appears that these free amino acids are involved in the ability of the estuarine bivalve Modiolus fluviatilis to osmoregulate in a wide range of salinities.

     

Activities of the three ammonia-forming enzymes in the tissues of the brackish-water bivalve Corbicula japonica

• 1.1. Activities of the three ammonia-forming enzymes, glutamate dehydrogenase, AMP deaminase and serine dehydrase (SerDH), were measured in tissues of gill, digestive diverticula, mantle and foot muscle of the brackish-water bivalve Corbicula japonica.
• 2.2. High levels of SerDH activity were detected in gill and digestive diverticula, while the activity levels of the other two enzymes were low.
• 3.3. The result suggests the significance of SerDH in amino acid degradation of this species.

     

Properties of hemocyanins isolated from Amazon river arthropods and molluscs

• 1. Hemocyanins from four organisms inhabiting the Amazon River were isolated and partially characterized.
• 2. Three arthropodan species (Dilocarcinus pagei cristatus, Silviocarcinus pardalinus andMacrobrachium amazonicum) possess hemocyanins whose subunit structure is remarkably simple. Regular and SDS polyacrylamide disc electrophoresis revealed predominantly single bands and no polymorphisms.
• 3. Oxygen-binding experiments showed that the three arthropodan hemocyanins possess large positive Bohr effects and pH dependence in the degree of subunit interaction.
• 4. The hemocyanin of one mollusc,Pila sp., was studied and its subunit size appears to be similar to that of other molluscan hemocyanins, i.e. a polypeptide of mol. wt 400,000. In the hemolymph,Pila hemocyanin probably exists as a mixture of 100 and 124S aggregates.
• 5. The oxygen binding properties of the large molecules ofPila hemocyanin are notable because of their low cooperativity and lack of a strong pH-dependence.

     

Biochemical variations in three species of prairie dogs (Cynomys)

• 1.1. Blood proteins were studied by polyacrylamide disc gel electrophoresis in three species of prairie dogs, Cynomys gunnisoni, C. leucurus, and C. ludovicianus.
• 2.2. The sera were separated into 13–15 fractions and the three species could be distinguished by both qualitative and quantitative differences in their serum patterns.
• 3.3. Qualitatively, variations in the occurrence and number of slow albumin fractions are diagnostic at the species leel.
• 4.4. Quantitative differences were most apparent in variation in the mobility of the major albumin fraction and the transferrin fraction.

     

Aerobic metabolism,octopine production and phosphoarginine as sources of energy in the phasic and catch adductor muscles of the giant scallop placopecten magellanicus during swimming and the subsequent recovery period

• 1.1. The energy contributions of aerobic metabolism, phosphoarginine, ATP and octopine in the adductor muscles of P. magellanicus were examined during swimming and recovery.
• 2.2. A linear relationship was observed between the size of the phosphoarginine pool and the number of valve snaps. A linear increase in arginine occurred during the same period.
• 3.3. Octopine was formed during the first few hours of recovery, particularly in the phasic muscle.
• 4.4. The restoration of the phosphoarginine pool appeared to be by aerobic metabolism.
• 5.5. It is concluded that the role of octopine formation is to supply energy when the tissues are anoxic and to operate at such a rate as to maintain the basal rate of energy production.