Identification and Characterization of a Novel Quadruple-Domain Galectin from the Hong Kong Oyster Crassostrea hongkongensis

Russian Journal of Marine Biology - Galectins, i.e. β-galactoside-binding animal lectins, play essential roles in the innate immunity systems of vertebrates and invertebrates. In the present...     

Isolation, characterization and inactivation of the mouse Mgat3 gene: the bisecting N-acetylglucosamine in asparagine-linked oligosaccharides appears dispensable for viability and reproduction

The biosynthesis of complex asparagine (N)-linked oligosaccharidesin vertebrates proceeds with the linkage of N-acetylglucosamine(GlcNAc) to the core mannose residues. UDP-N-acetylglucosamine:ß-D-mannosideß1–4 N-acetylglucosaminyltransferase III (GlcNAc-TIII,EC2.4.1.144) catalyzes the addition of GlcNAc to the mannosethat is itself ß1–4 linked to underlying N-acetylglucosamine.GlcNAc-TIII thereby produces what is known as a ‘bisecting’GlcNAc linkage which is found on various hybrid and complexN-glycans. GlcNAc-TIII can also play a regulatory role in N-glycanbiosynthesis as addition of the bisecting GlcNAc eliminatesthe potential for     

Genomic analysis of C-type lectins

Many biological effects of complex carbohydrates are mediated by lectins that contain discrete carbohydrate-recognition domains. At least seven structurally distinct families of carbohydrate-recognition domains are found in lectins that are involved in intracellular trafficking, cell adhesion, cell-cell signalling, glycoprotein turnover and innate immunity. Genome-wide analysis of potential carbohydrate-binding domains is now possible. Two classes of intracellular lectins involved in glycoprotein trafficking are present in yeast, model invertebrates and vertebrates, and two other classes are present in vertebrates only. At the cell surface, calcium-dependent (C-type) lectins and galectins are found in model invertebrates and vertebrates, but not in yeast; immunoglobulin superfamily (I-type) lectins are only found in vertebrates. The evolutionary appearance of different classes of sugar-binding protein modules parallels a development towards more complex oligosaccharides that provide increased opportunities for specific recognition phenomena. An overall picture of the lectins present in humans can now be proposed. Based on our knowledge of the structures of several of the C-type carbohydrate-recognition domains, it is possible to suggest ligand-binding activity that may be associated with novel C-type lectin-like domains identified in a systematic screen of the human genome. Further analysis of the sequences of proteins containing these domains can be used as a basis for proposing potential biological functions.     

ERRATA

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IDENTIFICATION AND EVALUATION OF THE FLAVOR-SIGNIFICANT COMPONENTS OF GINGER ESSENTIAL OIL

The flavorfully significant compounds in ginger essential oilwere selected statistically by use of a step-wise multiple regressionanalysis which treated individual peak quantitities on a gaschromatogram as independent variables and taste panel scoresfor ginger flavor intensity as dependent variables. The statisticalanalysis showed that four gas chromatographic peaks, consistingof a terpineol, citral a, citral b (peak 11), ß-sesquiphellandrene,ar-curcumene (peak 14), nerolidol (peak 19), and a sesquiterpenealcohol (peak 29), accounted for 85% of the panel's flavor response(R²=0.85). Taste panel evaluations of the isolated components indicatedthat ß-sesquiphellandrene and ar-curcumene are theprime contributors to the characteristic ‘ginger’attribute. -Terpineol, citral a and citral b contribute to the‘lemony’ attribute of ginger oil and may thereforebe desirable additives to whole ginger oil to intensify its‘lemony’ character. Nerolidol contributes to the‘woody’ or ‘soapy’ attribute and doesnot appear to be a good potential additive to ginger oil. A trained sensory panel judged a mixture of -terpineol, citrala, citral b, ß-sesquiphellandrene, ar-curcumene andnerolidol to be characteristic of ginger oil. The panel founda mixture of these chemicals, in combination with ginger heatchemicals, to be a suitable imitation ginger flavor. The study has shown that statistics can be used in selectingthe individual flavor contributing components of a flavor essence.This approach can greatly reduce the number of compounds whichneed be identified in the investigation of the composition ofnatural flavoring substances. ^* Scientific Article No. A1996, Contribution No. 4938, MarylandAgricultural Experimental Station (Food Science Program).     

Factors Affecting the Abscission of Reproductive Organs in Yellow Lupins (Lupinus luteus L.): III. ENDOGENOUS GROWTH SUBSTANCES IN VIRUS-INFECTED AND HEALTHY PLANTS AND THEIR EFFECT ON ABSCISSION

Extracts of small and mature-size lupin pods yielded four substancesaffecting the growth of wheat-coleoptile sections: one acidpromotor (A), two acid inhibitors(B and X), and one neutralinhibitor(Y). Inhibitor B was extremely active, however, coleoptile sectionsshowed no signs of toxic effects; they resumed growth at a rapidrate after rinsing them and adding ß-indolylaceticand (IAA) to the medium. 1 µg of IAA was required to counteractthe effect of ‘B’ extracted from 230 mg. Of tissue.On an equal fresh weight basis the inhibiting action of ‘B’in lupin pods was 500–1,500 times more potent than thatof ‘inhibitor ß’ in etiolated pea seedlings. Small pods of plants infected with pea-mosaic virus yielded3 times the amount of ‘A’ of healthy plants (equivalentto 1 µg. IAA 0.3 µg. IAA per 25 g. of tissue respectively),and approximately the amount of ‘B’. Mature podsof virus-infected plants again yielded more‘A’,but also 2? times more ‘B’ than pods of healthyplants. Healthy pods yielded more ‘A’ than virus-infectedpods, and there was no difference in ‘X’. A lupin abscission test was developed and the effects of proximaland distal application of -naphthyl acetic acid (NAA) are presented,and discussed with respect to results of other abscission tests. ‘A’ accelerated abscission when applied proximally,and delayed or prevented it when applied distally. ‘B’strongly accelerated abscission when applied in either way.A possible mechanism explaining the abscission-inducing effectof developing pods on later flowers is discussed in terms ofthe substances ‘A’ and ‘B’. The partlyprevented abscission observed on virus-infected plants was foundto agree well with the proposed mechanism.     

Comparative Immunology

Comparative Immunology includes an evolutionary approach toimmunity revealing the immune system as widespread and necessaryfor survival. Although many immunology textbooks and coursesare often oriented solely toward medicine orallied professions,comparative immunology is a unique beginning for advanced undergraduatestudents of biology, zoology, and immunology. A more biologicapproach to immune competence causes mammalian immunologiststo view comparative immunology as an introduction to numerousectothermic vertebrates and invertebrates. Such exotic animalsare sources of meaningful facts important for anyone viewingsignificant breakthroughs in immunology in phylogenetic perspective.Animals preserve their unique individuality by distinguishingbetween self and non-self to protect against infection and possibleextinction. The earliest beginnings of immune reactions arebest exemplified among the invertebrates by recognition, phagocytosis,graft rejection accompanied by specificity and weak memory,and the induction of agglutinins and lectins. Although vertebratesshare these same responses, the main differences include moreextensive evidence of specificity and memory and the uniquecapacity to synthesize immunoglobulins. Acknowledging the problemof recognition as the common basis for the immune response,certain members of the Ig superfamily, notably Thy-1 and ß₂-microglobulin,will help our attempts to define evolutionary pressures thatcaused the development of immunity.     

The ubiquity of the insulin superfamily across the eukaryotes detected using a bioinformatics approach

The insulin superfamily is composed of a diverse group of proteins that share a common structural design whose most notable feature is a set of disulfide bonds. There is now sufficient experimental and bioinformatics evidence that it is represented in at least a number of well-investigated invertebrates, where they have been found to intervene mainly in complex processes such as mitosis, cell growth, castes differentiation, and fertility. In this article we automated a methodology first proposed elsewhere-that combines sequence similarity with assessing membership to the superfamily by conservation of structuraly key residues-to identify putative insulin-like peptides (ILPs) in completely sequenced genomes, and applied it as a pipeline to a group of 46 organisms both vertebrates and invertebrates. As a result, we were able to identify 1,653 putative members of the insulin superfamily, from 17 putative members in C. savigny to 58 in X. tropicalis. Moreover, we found that structural distinctions-such as peptides length-between functionally diverse members of the superfamily found in vertebrates, that is, insulins, IGFs, and relaxins, are not equally represented in invertebrates genomes, suggesting that such divergence has occurred only recently in the evolutionary history of vertebrates.     

The nicotinic acetylcholine receptor gene family of the nematode Caenorhabditis elegans: an update on nomenclature

The simple nematode, Caenorhabditis elegans, possesses the most extensive known gene family of nicotinic acetylcholine receptor (nAChR)-like subunits. Whilst all show greatest similarity with nAChR subunits of both invertebrates and vertebrates, phylogenetic analysis suggests that just over half of these (32) may represent other members of the cys-loop ligand-gated ion channel superfamily. We have introduced a novel nomenclature system for these “Orphan” subunits, designating them as lgc genes (ligand-gated ion channels of the cys-loop superfamily), which can also be applied in future to unnamed and uncharacterised members of the cys-loop ligand-gated ion channel superfamily. We present here the resulting updated version of the C. elegans nAChR gene family and related ligand-gated ion channel genes.     

Biological Responsiveness to Pheromones Provides Fundamental and Unique Insight into Olfactory Function

When exposed to the odor of conspecifics, most organisms exhibitan adaptive behavioral response, particularly if the individualsare sexually mature. Evidence increasingly suggests that behavioralresponsiveness to these odors, which are termed ‘pheromones’,reflects neuroethological mechanisms associated with olfactoryfunction. Reproductive pheromones, which are the best understood,are commonly used by both invertebrates and vertebrates. Inboth instances they are generally comprised of mixtures of compoundsand behavioral responsiveness to them is largely instinctual,sexually-dimorphic, and attributable to a specialized component(s)of the olfactory system. While pheromonal responsiveness insome systems (e.g. moths) appears highly stereotypic and symptomaticof a relatively simple ‘labeled line’, behavioralresponsiveness of other animals (e.g. rodents) can be modifiedby experience, suggesting a more complex underlying centralmechanism. In any case, our understanding of these fascinatingsystems is progressing only because of an active dialogue betweenbehavioral and neurological investigations. This review brieflyexamines how behavioral studies have provided fundamental insightinto the neuroethology of olfactory function by drawing comparisonsbetween some of the better understood sex pheromone systemswhich have been described in heliothine moths, the goldfish,and the pig. Many similarities between invertebrate and vertebratepheromone systems are noted. Chem. Senses 21, 245–256,1996.     

Specific {beta} glucanases as tools for polysaccharide structure determination

A number of interesting ß-glucanases with a varietyof substrate specificities have recently been described. Theseenzymes are ideal tools for elucidating the structure of thewide array of glucose polymers found in nature. Since many ofthe enzymes can be easily produced in substantial quantitiesby cloning and ‘overexpression’ techniques, theyare available for other purposes as well. For example, theymay substantially increase the nutritional value of grain andother plant materials. glucanases ß-glucans polysaccharides     

Behavioral responses of the brittle star Ophiura ophiura to amino acids, acetylcholine and related low-molecular-weight compounds

Feeding behavior of the brittle star Ophiura ophiura includesorienting posture, orienting movements, arm ‘walking’,changing the direction of ‘walking’ arm coilingand ingestion. All sequential behavior patterns were releasedor enhanced by single low-molecular-weight compounds. Stimuliwhich released ‘walking’ behavior at high concentrations(10^–4 M) in all the test animals are listed in decreasingorder of sensitivity: sarcosine, glycine, urea, L-valine, L-leucine,L-methionine, L-homocysteine, L-norvaline, L-norleucine, L-threonine,L-serine, S-methyl-L-cysteine, L-proline. Threshold values forsingle amino acids were as much as 100 times different in differentindividuals and ranged from 3 x 10^–9 to 3 x 10^–7M for the most effective stimulus, sarcosine, and from 10^–6to 10^–4 M for proline. Above 10^–5 M, only L-prolineregularly released a second behavior pattern, the arm coilingresponse, which temporarily inhibited the ‘walking’behavior. Behavioral thresholds for the ‘walking’behavior for L(+)-lactate and L-alanine were higher than thosefor the orienting movements. Thyoglycolic acid and ß-alaninereleased tube feet walking, which is not part of the feedingbehavior. Structure—activity comparisons were studied at estimated10^–5 M concentrations. Gycline, sarcosine, L-valine, L-norvaline,L-leucine, L-isoleucine, DL-norleucine and DL-homocysteine releasedarm ‘walking’ behavior in more than 75% of all thetests. With the exceptions of S-methyl-and S-ethyl-cysteine,and glycine methylester, derivatives of amino acids were noteffective behavioral stimuli in Ophiura ophiura. L-Isomers ofvaline and leucine regularly stimulated the ‘walking’behavior while their D-isomers were effective in some testsand ineffective in others. Acetylcholine iodide, acetyl-ß-methylcholine chloride and choline phosphate chloride regularly released‘walking’ behavior at concentrations above 10^–5M.     

Xenopus laevis L-14 lectin is expressed in a typical pattern in the adult, but is absent from embryonic tissues

Soluble, dimeric, lactose-binding lectins with subunit Mr ofñ 14–16 x 10³, here called L-14s, are expressedin multiple tissues in all vertebrates that have been examined.L-14s have particular affinity for polylactosamine chains onlaminin, co-localize with laminin in some basement membranes,and influence adhesion to laminin and proliferation for somecultured cells. In previous studies of mammals and chickens,L-14s have been found at high levels in a variety of adult tissues,such as muscle and peripheral nerve, but at much higher levelsin many embryonic tissues, suggesting a special role in development.To further explore possible roles of L-14 in embryogenesis,we have studied the expression of L-14 in embryonic and adultXenopus laevis tissues. Except for the abundant L-14 in poisonglands in adult Xenopus skin, we find that Xenopus L-14 is expressedin the same general distribution as its mammalian homologues.However, we could detect no expression of L-14 in Xenopus embryosusing either a sensitive immunoassay for the protein or a sensitiveRNase protection assay for its mRNA. Furthermore, use of affinitychromatography to identify other lactose-binding lectins inembryonic tissue revealed only scarce proteins with higher subunitmolecular weights. These results suggest that in X.laevis L-14functions in adult tissues and is not involved in embryogenesis. ß-galactoside-binding lectin L-14 lectin Xenopus laevis.     

Concentration-dependent changes of perceived odor quality

In order to assess the dependence of perceived odor qualityon odorant concentration, we studied 21 subjects. For eightsubjects all possible pairs from a pool of six odorants at threedecimal dilutions were presented, and subjects were requestedto state whether members of the pair were qualitatively ‘similar’or ‘different’ It was found that while pairs withthe same odorant at identical concentrations were judged ‘similar’in >90% of the cases by all subjects, scores went down to10% ‘similar’ judgements in some cases when thesame odorant was presented at a 100-fold concentration difference.Large time-invariable differences were found among subjectsand among odorants. For the additional 13 subjects, all possiblepairs from a pool of four odorants at three decimal dilutionswere presented. Subjects were instructed to state whether membersof the pair were qualitatively ‘same’ or ‘different’,and were also requested to rank the degree of difference ona visual analogue scale. Results for this group were, in general,similar to the results of the former group of subjects and goodagreement between the two tasks was found. The results suggestthat variations in olfactory stimulus magnitude may be perceivedas quality differences, as previously shown for vision and audition.     

Some Early Responses of Helianthus annuus L. to Flooding: I. THE EFFECTS OF FLOODING ON THE UPTAKE AND LEAKAGE OF 'NON-ELECTROLYTES' BY ROOTS

Drakeford, D. R., Mukherjee, I. and Reid, D. M. 1985. Some earlyresponses of Helianthus annuus L. to flooding. I. The effectsof flooding on the uptake and leakage of ‘non-electrolytes’by roots.—J. exp. Bot. 36: 1705–1715. The object of this work was to examine some of the early effectsof flooding on roots. A hydroponic system was developed thatgave good control over watering, degree of oxygenation of thebathing medium and allowed measurement of short term changesin the composition of the bathing medium. Excised roots, floodedfor 24 h, were shown to take up less [³H) ß-alaninethan non-flooded roots and also leaked more [³H] ß-alanineinto a distilled water bathing medium. Further, flooded excisedroots lost more protein to the bathing medium, with ‘young’(5–7 d) roots showing greater losses than ‘old’(11–14 d) roots. However, young roots had more proteinin the tissue even after greater loss. Young roots remainedhealthier and lost less fresh weight than old roots. Abscisicacid was shown to have a small role in protecting ‘young’roots from the effects of flooding. Key words: ABA, abscisic acid, anaerobic, flooding, leakage, roots, uptake, waterlogging     

Emission of Volatiles From Brown Boronia Flowers: Some Comparative Observations

Extensive research has focused on the concentration of aglyconeswithin brown boronia (Boronia megastigma) flowers, however emissionof volatiles into the headspace above these flowers is not welldocumented. Using solid-phase microextraction (SPME) to trapvolatiles and GCMS analysis, we observed 23 volatiles in theheadspace above buds and flowers throughout flower maturation,above dissected floral organs and above whole plants held for36 h under either continuous light, continuous dark or 12 hlight:12 h dark:12 h light treatments. Fully-opened flowersemitted the most complex mixture of volatiles and in the greatestquantity, with a rapid decline in senescent flowers. Caryophyllene,humulene and bicyclogermacrene declined as flower buds matured;ß-ionone increased. From the individual floral organs,emission from the petaline anthers comprised 38% of total emissionsfrom the (calculated) ‘whole flower’, with 27% contributedby the petals and 10.5% by the stigma. Monoterpenes dominatedthe headspace from the calyx; dodecyl acetate, methyl jasmonateand (Z)-n-heptadec-8-ene were relatively predominant in emissionsfrom the androecium. ß-Ionone, the major floral volatilein brown boronia, dominated volatiles emitted from the stigma(87%). However, the relatively tiny petaline anthers, activein pollen production and high in carotenoids, contributed thegreatest overall amount of ß-ionone to emission fromthe whole flower. There were three different patterns in emissionof volatiles from plants in response to different light conditions:(1) emission patterns identical irrespective of light environment,with maximum emission in the ‘endogenous’ dark period,i.e. when the plant would normally have been in the dark (-pinene);(2) similar emission in all treatments, with an increase anddecline over a period of 26 h (5-acetoxy linalool, cyclic ß-ionone,dodecyl acetate and (Z)-n-heptadec-8-ene); and (3) emissionin all treatments but enhanced in the dark, with a 27.5 h periodin some cases (cyclic ß-ionone endoperoxide, dihydroß-ionone, ß-ionone, and ‘total volatiles’).Preliminary evidence is presented for endogenous control ofemission of a number of volatiles such as -pinene, with perhapsdiurnal control of others such as ß-ionone. Copyright2000 Annals of Botany Company Boronia megastigma, brown boronia, SPME, headspace, floral volatiles, ß-ionone     

Transient Expression of Exogenous DNA in Intact, Viable Wheat Embryos Following Particle Bombardment

Expression of foreign DNA has been detected in intact, germinatingwheat embryos (Triticum aestivum L.) following bombardment withtungsten particles complexed with a reporter gene encoding thebacterial enzyme ß-glucuronidase (‘GUS’:E.C.3.2.1.31). Expression was detected in situ in individualcells and groups of cells, by supplying the germinating embryoswith the chromogenic substrate of the GUS enzyme, ‘X-gluc’.Expression was dependent on the presence of a constitutive plantpromoter, the Cauliflower Mosaic Virus ‘35S’ promoter,fused to the GUS structural coding sequence. The relative simplicityof this technique recommends its future use for the assay ofregulatory elements which control the spatial and temporal specificityof genes expressed during embryo development. Key words: Transient expression, particle bombardment, wheat embryo, Triticum aestivum L.     

Lectin binding by macroglial and microglial cells in the brains of young normal and myelin-deficient (md) mutant rats

To distinguish macroglia (oligodendrocytes and astrocytes) frommicroglia in the brain, ‘markers’ that have beenused in previous studies include carbonic anhydrase II (CAII)immunoreactivity for macroglia, and Ricinusand Bandeiraea(BS-I)lectins for microglia. However, in rats <1 week of age, manycells stained intensely with both anti-CAII and the labelledBS-I lectin. If some of the BS-I+/CAII+ cells were macroglia,and not microglia, BS-I should no longer be regarded as specificfor microglia. To confirm or rule out that possibility, lectinhistochemistry and double immunofluorescence staining were performedin tissue from the brains of normal young rats and from themicroglial cell-enriched brains of myelin-deficient mutant rats.BS-I+/ CAII+ cells were found and examined. The BS-I +/CAII-cellsresembled macrophages and microglia and did, indeed, differin sizes and shapes from the BS-I+/+cells. The BS-I+/CAII+ cellsappeared to represent CAII+ putative oligodendrocyte precursorsdescribed previously. Although less obvious, a lectin-bindingstructure was also observed in astrocytes. Lectins may cross-reactwith macroglial glycoproteins. For example, a glycoprotein foundin o ligodendrocytes and myelin, the myelin-associated glycoprotein(MAG), is related to the Ig superfamily and cell adhesion molecules.Therefore, it is cautioned that lectins and antibodies againstmembers of the latter families of proteins should be used ascell-type specific markers only if other parameters are alsoexamined. astrocytes carbonic anhydrase lectins microglia oligodendrocytes     

Uptake and Metabolism of Sugars by Suspension cultured Catharanthus roseus Cells

p. 186, right column line 11 ‘27.2 KBq’ change to‘37.2 MBq’ p. 187, left column line 8 ‘27.2 KBq’ change to‘37.2 MBq’ line 10 ‘13.6 KBq’ change to ‘18.6 MBq’ line 11 ‘13.6 KBq’ change to ‘18.6 MBq’ line 21 ‘89%’ change to ‘80%’ right column line 24 ‘CLC-NH₄’ change to ‘CLC-NH₂’ P. 189, Table 1 appeared incorrectly: it should appear as indicated.     

Protein phylogenies provide evidence of a radical discontinuity between arthropod and vertebrate immune systems

 Protein phylogenies were used to test the hypothesis that aspects of the innate immune system of vertebrates have been conserved since the last common ancestor of vertebrates and arthropods. The phylogeny of lysozymes showed evidence of conservation of function, but phylogenies of seven other protein families did not. Natural resistance-associated macrophage protein, nitric oxide synthetase, and serine protease families all showed a pattern of gene duplication within vertebrates after their divergence from arthropods, giving rise to immune system-expressed genes in vertebrates. Insect hemolin, a member of the immunoglobulin superfamily, was found not to be closely related to members of that family having an immune system role in vertebrates; rather, it appeared most closely related to both arthropod and vertebrate molecules expressed in the nervous system. Thus, hemolin seems to have evolved its role independently in insects, probably through duplication of a neuroglian-like ancestor. Furthermore, vertebrate immune system-expressed serpins, chitinases, and pentraxins were found to lack orthologous relationships with arthropod members of the same families also functioning in immunity. Therefore members of these families have evolved immune system functions independently in the two phyla. It is now widely recognized that the specific immune system of vertebrates has no counterpart in invertebrates; these phylogenetic analyses suggest that there is a similar evolutionary discontinuity with respect to innate immunity as well. Received: 10 May 1997 / Revised: 10 September 1997