Distribution of catecholamines,indoleamines, and their precursors and metabolites in the scallop,Placopecten magellanicus (Bivalvia,Pectinidae)

Summary 1. Although monoamines are well-known to play important roles in molluscan physiology, we are far from fully understanding the synthetic and degradative pathways of these substances, particularly in commercially important bivalve species. In the present study endogenous catecholamines, indoleamines, and their possible precursors and metabolites were detected in the scallop,Placopecten magellanicus, by high-performance liquid chromatography coupled to electrochemical detection.2. Chromatographic analysis of CNS (cerebral, pedal, and parietovisceral combined), gill, gonad, kidney, mantle, liver, heart, fast adductor muscle, and foot disclosed the presence of the catecholamines 3,4-dihydroxyphenylalanine, dopamine, norepinephrine, and epinephrine and their metabolites normetanephrine, metanephrine, 3,4-dihydroxyphenylacetic acid, and homovanillic acid.3. Dopamine was detected most frequently and most consistently among all catecholamines. The concentrations of dopamine (1400 pg/mg wet weight) and its major metabolite 3,4-dihydroxyphenylacetic acid (300 pg/mg wet weight) were highest in the CNS. Following the CNS, dopamine was also abundant in other tissues such as heart, foot, and gill. The concentration of norepinephrine (1000 pg/mg wet weight) was highest in the CNS followed by the heart (700 pg/mg wet weight) and gill (600 pg/mg wet weight).4. The indoleamine, 5-hydroxytryptamine, was present in considerable amounts in all tissues, but its content was highest in the foot (2700 pg/mg wet weight) followed by the CNS (1150 pg/mg wet weight) and gonad (1000 pg/mg wet weight). The precursor 5-hydroxytryptophan was also abundant in the foot followed by the gonad, CNS, and heart.5. The oxidative metabolite 5-hydroxy-3-indole acetic acid was detected in the largest amount in CNS (200 pg/mg wet weight), whereasN-acetyl-5-hydroxytryptamine was detected in trace amounts in CNS, gonad and foot. This study also presents evidence for -glutamyl dopamine and -glutamyl 5-hydroxytryptamine as the possible alternate catabolic products of dopamine and 5-hydroxytryptamine, respectively, as previously described in gastropods.6. Thus, the detection of monoamines and their precursors and metabolites in scallop strongly suggests the presence of mammalian-type enzymic action of hydroxylation, oxidation, and methylation pathways leading to synthesis and degradation of detected compounds. Furthermore, this is the first study to disclose the evidence of nonconventional metabolic pathways for dopamine (-glutamyl dopamine dopamine dihydroxyphenylacetic acid homovanillic acid) and 5-hydroxytryptamine (-glutamyl 5-hydroxytryptamine 5-hydroxytryptamine 5-hydroxy-3-indoleacetic acid) inactivation in a bivalve species.     

Detection of the gangliosideN-glycolyl-neuraminyl-lactosyl-ceramide by biotinylatedEscherichia coli K99 lectin

K99 lectin fromEscherichia coli was purified and biotinylated via its carboxyl groups using biocytin hydrazide and a water soluble carbodiimide. Biotinylation of two out of the nine carboxyl groups was sufficient to permit detection of the lectin by avidin and did not cause any loss of the haemagglutinating activity. It was demonstrated that the biotinylated K99 lectin retained other important properties of native K99 and that it will probably become a very sensitive detecting reagent. Indeed, it was able to bind to HeLa cells, as do intact bacteria carrying K99 fimbriae, and also to recognizeN-glycolyl-neuraminyl-lactosyl-ceramide in an overlay binding assay. Abbreviations: NeuAc,N-acetylneuraminic acid; NeuGc,N-glycolylneuraminic acid; PBS, phosphate buffered saline (0.9% NaCl containing 150mm sodium phosphate, pH 7.2); LPS, lipopolysaccharide; BCHZ, biocytin hydrazide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; BSA, bovine serum albumin; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; DMEM, Dulbecco's modified Eagle medium. For the gangliosides, trivial names and structures are given according to the recommendations in [43]. NeuAc2-3Gal1-4Glc1-1Cer (NeuAc-GM₃); NeuGc2-3Gal1-4Glc1-1Cer (NeuGc-GM₃); GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer (GM₂); NeuAc2-8NeuAc2-3Gal1-4Glc1-1Cer (GD3); Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer (GM₁); NeuAc2-3Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer (GD_1a); Gal1-3GalNAc1-4(NeuAc2-8NeuAc2-3)Gal1-4Gle1-1Cer (GD_1b); NeuAc2-3Gal1-3GalNAc1-4(NeuAc2-8NeuAc2-3) Gal1.-4Glc1-1Cer (GT_1b). NeuGc2-3Gal1-4GleNAc1-4Gal1-4Glc1-1Cer (NeuGc-SPG).     

Ultrastructure of lycopene containing chromoplasts in fruits ofAglaonema commutatum schott (Araceae)

Summary The ripening process in fruits ofA. commutatum is characterized by clearly distinguishable developmental stages of their pericarp plastids. With respect to predominant pigments and ultrastructural features the following scheme is proposed: 1. The green stage with a tendency to thylakoid degeneration and plastoglobule formation leads to 2. the yellow stage. An increasing number of globules, mostly being membrane associated, are converted to tubules. In this stage, the main pigments are -carotene and cryptoxanthine. The development of membraneous invaginations from the inner plastid envelope leads to 3. the red stage. Concomitantly with lycopene synthesis and incorporation, these envelope-derived membranes expand and become electron dense (after KMnO₄ treatment), but maintain their triple-layered structure. Chromoplasts of deep red coloured fruits (1,400 g lycopene g^–1 dry wt) contain lycopene crystals within the lumina of membraneous sacs which are also derived from the inner envelope membrane. The molar ratio between the three main pigments -carotene, cryptoxanthine, and lycopene is about 1110 in this final state. GA/OsO₄ fixation is unable to stabilize the lycopenic structures (membranes as well as crystals).     

Metal Cations Defibrillize the Amyloid Beta-Protein Fibrils

Amyloid beta-protein (A) is the major constituent of amyloid fibrils composing -amyloid plaques and cerebrovascular amyloid in Alzheimer's disease (AD). We studied the effect of metal cations on preformed fibrils of synthetic A by Thioflavin T (ThT) fluorescence spectroscopy and electronmicroscopy (EM) in negative staining. The amount of cross beta-pleated sheet structure of A 1–40 fibrils was found to decrease by metal cations in a concentration-dependent manner as measured by ThT fluorescence spectroscopy. The order of defibrillization of A 1–40 fibrils by metal cations was: Ca²⁺ and Zn²⁺ (IC₅₀ = 100 M) > Mg²⁺ (IC₅₀ = 300 M) > Al³⁺ (IC₅₀ =1.1 mM). EM analysis in negative staining showed that A 1–40 fibrils in the absence of cations were organized in a fine network with a little or no amorphous material. The addition of Ca²⁺, Mg²⁺, and Zn²⁺ to preformed A 1–40 fibrils defibrillized the fibrils or converted them into short rods or to amorphous material. Al³⁺ was less effective, and reduced the fibril network by about 80 % of that in the absence of any metal cation. Studies with A 1–42 showed that this peptide forms more dense network of fibrils as compared to A 1–40. Both ThT fluorescence spectroscopy and EM showed that similar to A 1–40, A 1–42 fibrils are also defibrillized in the presence of millimolar concentrations of Ca²⁺. These studies suggest that metal cations can defibrillize the fibrils of synthetic A.     

Novel promoter and splice junction defects add to the genetic,clinical or geographic heterogeneity of β-thalassaemia in the Portuguese population

Summary In order to delineate the spectrum and the relative abundance of -globin gene defects causing thalassaemia in the Portuguese population, a representative sample was analysed including 51 -thalassaemia carriers along with 26 patients representing different clinical phenotypes. Seven mutations were identified, four of which [codon 39 (CT), 39%; intervening sequence (IVS)1 nucleotide (nt) 1 (GA), 26%; IVS1 nt 110 (GA), 17%; IVS1 nt6 (TC), 15%] account for 97% of 93 -thalassaemia chromosomes. Two previously undescribed mutations, namely a CT substitution at position — 90 in the proximal CACCC box, and the deletion of nucleotides 4 and 5 (AG) in IVS 2 were identified. The uncommon, though ubiquitous, GT transversion at codon 121 was found once upon haplotype V. Direct prenatal diagnosis can be offered to 95% of couples at risk of bearing a thalassaemic child.     

The adaptive significance of cultural behavior: Comments and reply

Summary Fundamentally, theoretically, there is only one process underlying genetic and cultural evolution: natural selection. Organism fitness-enhancement (adaptive significance) is one of its practical mechanisms; group formation and maintenance is another, often but not always through fitness-enhancement; and need-fulfillment is still another. If Durham can accept that formulation, and switch from organism-thinking to instruction-thinking (Cloak, 1975: 178), he will free himself from two handicaps: First, he can forget his worries about reductionism and determinism (1976a: 100, 101). Under this general theory of natural selection, cultural evolutionis biological evolution, continued by other (nongenetic) means. Second, he will spare himself the appearance of anthropomorphism, mentalism, and wishy-washiness attendant on his discussion of kinds of significance, other than adaptive significance, of cultural behaviors (1976a: 102–106, 115).     

Evolution of a protein superfamily: Relationships between vertebrate lens crystallins and microorganism dormancy proteins

Summary A search of sequence databases shows that spherulin 3a, an encystment-specific protein ofPhysarum polycephalum, is probably structurally related to the - and -crystallins, vertebrate ocular lens proteins, and to Protein S, a sporulation-specific protein ofMyxococcus xanthus. The - and -crystallins have two similar domains thought to have arisen by two successive gene duplication and fusion events. Molecular modeling confirms that spherulin 3a has all the characteristics required to adopt the tertiary structure of a single -crystallin domain. The structure of spherulin 3a thus illustrates an earlier stage in the evolution of this protein superfamily. The relationship of - and -crystallins to spherulin 3a and Protein S suggests that the lens proteins were derived from an ancestor with a role in stressresponse, perhaps a response to osmotic stress.     

Corticosteroid-binding globulin synthesis and distribution in rat white adipose tissue

The lactone isolated from Fusarium termed L659,699 is a potent specific inhibitor of the enzyme 3hydroxi3methylglutaril coenzyme A (HMG-CoA) synthase. In cultures of smooth muscle cells (SMC) isolated from aortic-arch of control (CSMC) and 5% of cholesterol diet (Ch-SMC) treated chicks, the incorporation of (¹⁴C)acetate to lipids (cholesterol, triacylglycerides and cholesterol ester) were greater in ChSMC cultures than in CSMC and the presence of 0.05 M L659,699 for 2 h in the incubation medium decrease the synthesis of cholesterol however the triacylglycerides synthesis increase. The effect of inhibitor is stronger in young cultures (3–4 steps) than in the older ones (11–12 steps). In young CSMC and ChSMC cultures the inhibition of cholesterol and triacylglycerides synthesis by L659,699 was reversal.     

Ultrastructure and behaviour of the spindle pole body of the aphid-pathogenic fungusErynia neoaphidis

Summary A detailed account of the ultrastructure and behaviour of the spindle pole body (SPB) of the entomophthoraceous fungusErynia neoaphidis is presented for the first time.The SPB consists of extranuclear (ENC) and intranuclear (INC) components. The ENC is a saucepan-shaped structure which lies in a pocket of the nuclear envelope. It is composed of a forked, fibrillar handle and a shallow, cylindrical pan. The pan has a wall of two layers, both of which are thickened with a regular periodicity so that they appear to be beaded. It is postulated that the pan is formed from rough endoplasmic reticulum and that it synthesizes the amorphous, electron-dense material coating the ENC.The INC is a saucer-shaped, electron-dense plaque in which the ends of the spindle microtubules terminate. During metaphase, a clear zone separates the INC from the nuclear envelope and persists until telophase. The roles of the amorphous, electron-dense material and the clear zone as well as the method of SPB replication are discussed.     

Carotenoid pigments of Sorangium compositum (Myxobacterales) including two new carotenoid glucoside esters and two new carotenoid rhamnosides

Summary The carotenoid pigments of the myxobacterium Sorangium compositum were analyzed by chromatographical and chemical techniques and by visible, infra red, and mass spectroscopy. Besides -carotene, neurosporene, torulene, lycopene, and 1,2-dihydro-1-hydroxy--carotene, four new carotenoid glycosides were found. These pigments were identified as 1,2-dihydro-1-hydroxy-torulene glucoside ester (I), 1,2-dihydro-3,1-dihydroxy-torulene glucoside ester (III), 1,2-dihydro-1-hydroxy-torulene rhamnoside (II), and 1,2-dihydro-3,1-dihydroxytorulene rhamnoside (IV).Fifth communication on the carotenoids of myxobacteria. Fourth communication see Arch. Mikrobiol. 76, 364–380 (1971).     

Liver and intestinal fatty acid binding proteins in control and TGFβ1 gene targeted deficient mice

The effect of transforming growth factor beta-1 (TGF1) expression on fatty acid binding proteins was examined in control and two strains of gene targeted TGF1-deficient mice. Homozygous TGF1-deficient 129 × CF-1, expressing multifocal inflammatory syndrome, had 25% less liver fatty acid binding protein (L-FABP) when compared to control mice. The decrease in L-FABP expression was not due to multifocal inflammatory syndrome since homozygous TGF1-deficient/immunodeficient C3H mice on a SLID background had 36% lower liver L-FABP than controls. This effect was developmentally related and specific to liver, but not the proximal intestine, where L-FABP is also expressed. Finally, the proximal intestine also expresses intestinal-FABP (1-FABP) which decreased 3-fold in the TGF1-deficient/immunodeficient C3H mice only. Thus, TGF1 appears to regulate the expression of L-FABP and I-FABP in the liver and the proximal intestine, respectively.Abbreviations L-FABP liver fatty acid binding protein - I-FABP intestinal fatty acid binding protein - TGF1 transforming growth factor beta-1 - TNF- tumor necrosis factor- - MIP- macrophage inflammatory protein- - PMSF phenylmethyl sulfonyl fluoride - PBS phosphate buffered saline     

Glycosphingolipids of leukocytes are unbranched at the galactopyranosyl residue and contain fucosylα-1-3-N-acetylglucosamine structures

Glycolipids of peripheral leukocytes which had been used for the production of interferon were separated into oligoglycosylceramides, polyglycosylceramides and polyglycosylpeptides (erythroglycan). Neutral oligoglycosylceramides comprised glucosylceramide, galactosylceramide, lactosylceramide, lactotriaosylceramide, globotriaosylceramide andneolactotetraosylceramide. Globotetraosylceramide was not detected. Glycolipids which were more complex thanneolactotetraosylceramide belonged exclusively to theneolacto series of compounds and were essentially unbranched at galactopyranosyl residues. The polyglycosylceramide fraction contained a glycolipid with a probable structure Gal1-4(Fuc1-3) GlcNAc1-3Gal1-4GlcNAc1-3 Gal1-4GlcNAc1-3Gal1-4Glc1-1ceramide. Polyglycosylpeptides were found only in trace amounts and were also unbranched at galactopyranosyl residues. All glycoconjugates studies did not contain significant amounts of carbohydrate structures derived from ABH immunodominant groups.Nomenclature Gal1-4Gal1-4GlcCer Lactotrioasylcermide (LcOse₃Cer) - Gal1-4Gal1-4GlcCer globotriaosylceramide, (GbOse₄Cer) - GalNAc1-3Gal1-4 Gal1-4GlcCer globoside (globotetraosylceramide, GbOse₄Cer) - Gal1-4GlcNAc1-3Gal1-4GlcCer paragloboside (lacto-N-neo tetraosylceramide,nLcOse₄Cer)     

Preliminary Study of Vertebral Growth Rings in the Whale Shark, Rhincodon typus, from the East Coast of South Africa

Growth rings (GR) in vertebral centra of 15 whale sharks, Rhincodon typus, four female (418–750cm precaudal length), 10 male (422–770cm), and one of unknown sex (688cm), were examined using x-radiography. GR counts were made from scanned images and count precision was determined using the average percentage error index (4.19%) and the index of precision D (3.31%). In females, counts ranged from 19 GR (418cm) to 27 GR (750cm); in males from 20 GR (670cm) to 31 GR (770cm). Three mature males had 20 GR (670cm), 24 GR (744cm) and 27 GR (755cm). A female with 22 GR (445cm) was adolescent. There was a linear relationship between centrum dorsal diameter and body length, and back-calculated body lengths at number of GR are presented. A linear relationship between body length and number of GR prevented the calculation of von Bertalanffy parameters from either observed or back-calculated values.     

Identification and characterization of the functional alpha origin of DNA replication of the R6K plasmid and its relatedness to the R6K beta and gamma origins

Summary The functional R6K origin is composed of two DNA elements, one of 580 bp carrying the origin sequences and the other of 277 bp containing the seven 22 bp direct repeats previosly identified as also required for and origin activity. These two genetic elements are separated by approximately 3,000 bp of R6K sequences which are dispensable for origin activity. The function of the origin depends on the presence in cis of the 580 bp and the 277 bp fragments and requires that they be oriented as in the intact R6K. Activation of the origin depends on the R6K replication initiation protein .Within the 580 bp of the origin, there is a sequence of 98 bp which appears as an inverted repeat of 96 bp in the replicon. Deletion of the 96 bp or 98 bp results in inactivation of the and the origins respectively. These long repeats are palindromic and it is suggested that these may serve as the recognition signals for initiation of DNA replication in the and the origins of R6K. DNA homology analysis performed on , and origin sequences, also reveals 10–23 bp sequences in the and the origins that are related to the family of 22 bp direct repeats in the origin which were shown previously to be binding sites for the protein.     

Long-term continuous reaction of immobilized β-fructofuranosidase

Summary -Fructofuranosidase was immobilized by alginate gel at high efficiency (92 %). The extreme long-term continuous reaction (half-life, 275 days) was achieved by the immobilized enzyme using sucrose at high concentration (500 mg ml^–1) to produce fructo-olicosaccharides, such as 1-kestose (Fru21Fru21aGlc) and nystose (Fru21Fru21Fru21aGlc).     

Partial mispairing and crossing-over between β0 and δ genes as the origin of the δ β0 thalassemia gene

Summary Two double heterozygous ⁰/⁰ thalassemic sibs of Mexican descent were studied. The father had a ⁰/⁰ genotype, while the mother, one sib and several maternal relatives were ⁰/⁰ heterozygotes. Parental consanguinity and an apparently low frequency of thalassemia among Mexicans suggested a possible common origin of both ⁰ and ⁰ genes. A hypothesis to explain such a possibility is proposed on the basis of a partial mispairing between ⁰ and genes followed by a crossing-over which would results in a ⁰ recombinant gene. This hypothesis could also be extended to explain the 22 gluala, 22 alaglu and 116 arghis Hb variants as recombinants from double crossing-over between and mispaired genes for which the name interstitial-Lepore is proposed.     

A new enzymatic route to the synthesis of 12-ketoursodeoxycholic acid

Summary Cholic acid (3,7,12-trihydroxy-5-cholanoic acid) was completely and selectively transformed into 12-ketoursodeoxycholic acid (3,7-dihydroxy-12-oxo-5-cholanoic acid) by means of two consecutive enzymatic steps catalyzed, the first, by 7- and 12-hydroxysteroid dehydrogenase and, the second, by 7-hydroxysteroid dehydrogenase. Coenzyme regeneration was carried out with -ketoglutarate-glutamate dehydrogenase and glucose-glucose dehydrogenase, respectively.