Intrinsic fluorescence studies on saccharide binding toArtocarpus integrifolia lectin

The combining region ofArtocarpus integrifolia lectin has been studied by using the ligand-induced changes in the fluorescence of the lectin. The saccharide binding properties of the lectin show that C-l, C-2, C-4, and C-6 hydroxyl groups of D-galactose are important loci for sugar binding. The -anorner of galactose binds more strongly than its -counterpart. Inversion in the configuration at C-4 as in glucose results in a loss of binding to the lectin. The C-6 hydroxyl group is also presumably involved in binding as D-fucose does not bind to the lectin.The lectin binds to the Thomsen-Friedenreich antigen (Gal(13)GalNAc) more strongly than the other disaccharides studied, viz. Gal/ (14) Gal and Gal (13) GlcNAc, which are topographically similar to T-antigen. This observation suggests that the combining region ofArtocarpus lectin is complementary to that of T-antigen.Solvent accessibility of the protein fluorophores have been probed by the quenching of protein fluorescence by Iodide ion in the absence and presence of sugar. In the presence of sugar a slight inaccessibility of the fluorophores to the solvent has been observed.Abbreviations MeGal 1-O-methyl--glucopyranoside - MeGal 1-O-methyl--glucopyranoside - GalNAc 2-acetamido-2-deoxy-galactose - Gal Galactose     

Investigation of the Relationship between the Lectin Activity of Azospirilla and Their α-Glucosidase, β-Glucosidase,and β-Galactosidase

The activities of -glucosidase, -glucosidase, and -galactosidase were studied during the isolation and purification of lectins from Azospirillum brasilenseSp7 and Azospirillum lipoferum59b cells. These enzymatic activities were revealed in crude extracts of surface proteins, protein fraction precipitated with ammonium sulfate or ethanol–acetone mixture, and protein fraction obtained by gel filtration on Sephadex G-75. The distribution of the enzymes between different protein fractions varied for the azospirilla studied. The cofunction of the A. brasilenseSp7 lectin and -galactosidase on the cell surface is assumed. A strong interaction between the A. lipoferum59b lectin and glucosidases was revealed. The lectin from A. lipoferum59b may possess saccharolytic activity.     

Myosatellite cells associated with different muscle fibre types in the Atlantic hagfish (Myxine glutinosa,L.)

Summary The incidence of myosatellite cells associated with white and red muscle fibres of the parietal muscle and red fibres of the craniovelar muscle was estimated by quantitative electron microscopy in the Atlantic hagfish (Myxine glutinosa, L.). Myosatellite cell nuclei constitute 3, 11 and 23 % of the total number of nuclei inside the basal lamina of the three types of muscle fibres, respectively. However, the total number of nuclei is highest in white fibres, most of the nuclei belonging to striated muscle cells. Myosatellite cell profiles in transverse sections constitute 23, 41 and 61 % of the number of muscle fibre profiles of the three types, respectively. The intervals between adjacent myosatellite cells are 135 m in white fibres, 55 m in red parietal fibres, and only 25 m in craniovelar fibres. Since craniovelar fibres are also comparatively thin, myosatellite cells constitute a significant fraction of the volume inside the basal lamina in these fibres. The myosatellite cells are 30–50 m long and up to 5 m thick. Some myosatellite cells possess few organelles, whereas others appear to contain many free ribosomes, granular endoplasmic reticulum, prominent Golgi apparatus and lysosome-like bodies.This investigation was supported by the Norwegian Research Council for Science and the Humanities (NAVF grant No. C20.30–37). The authors are indebted to Jorunn Line Vaaland and Berit Branil for technical assistance, and to Dr. Finn Walvig, Biological Station, University of Oslo, Drøbak, for supplying the hagfish     

DNA polymorphisms in the ψβ1- and β-globin gene regions in asian macaques

DNA polymorphisms in the ₁--globin gene region in nine Asian macaques(Macaca fuscata, M. mulatta, M. nemestrina, M. cyclopis, M. fascicularis, M. arctoides, M. radiata, M. maura, andM. assamensis) were examined using several restriction endonucleases and the human ₁, IVS2, and IVS2 probes. TheBamHI site 3 to the -globin gene was polymorphic inM. fuscata andM. mulatta, while the HincII site and the EcoRI site in the ₁-globin gene region was highly polymorphic inM. fuscata andM. mulatta, respectively. These polymorphic sites also seem to be present in other Asian macaques. The present study of the polymorphism at theBamHI site 3 to the -globin gene in Asian macaques supports, at the nuclear DNA level, the idea that thefascicularis group includingM. fuscata, M. mulatta, M. cyclopis, andM. fascicularis is different from other Asian macaque groups.This study was supported in part by the Cooperation Research Program of the Primate Research Institute, Kyoto University.     

A fluorometric assay of ceramide glycanase with 4-methylumbelliferyl β-d-lactoside derivatives

4-Methylumbelliferyl 6-O-benzyl--d-lactoside (6Bn-MU-Lac) and some related compounds were synthesizedvia different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, theK _m value is 0.232 mM at pH 5. Abbreviations: CGase, ceramide glycanase; Gal, galactose; Glc, Glucose; Lac, lactose; MU, 4-methylumbelliferone; MU-Lac, 4-methylumbelliferyl -d-lactoside; bBn-Lac, 6-O-benzyl-lactose; 6Bn-MU-Lac, 4-methylumbelliferyl 6-Obenzyl--d-lactoside; 46Bd-MU-Lac, 4-methylumbelliferyl 4,6-O-benzylidene--d-lactoside; MU-Cel, 4-methylumbellifery -d-cellobioside; 46Bd-MU-Cel, 4-methylumbelliferyl 4,6-O-benzylidene--d-cellobioside; TLC, thin layer chromatography;¹H-NMR, proton nuclear magnetic resonance; GSL, glycosphingolipids; CSA, 10-camphorsulfonic acid. See Scheme 1 for chemical structures.     

Solution conformations of proline rings in proteins studied by NMR spectroscopy

Summary Three different conformations of proline rings in a protein in solution, Up, Down and Twist, have been distinguished, and stereospecific assignments of the pyrrolidine -, - and -hydrogens have been made on the basis of ¹H-¹H vicinal coupling constant patterns and intraresidue NOEs. For all three conformations, interhydrogen distances in the pairs -³, ³-³, ²-², ²-², and ³-³ (2.3 Å) are shorter than those in the pairs -², ²-³, ³-², ²-³, and ³-² (2.7–3.0 Å), resulting in stronger NOESY cross peaks. For the Up conformation, the ³-² and ²-³ spin-spin coupling constants are small (<3 Hz), and weak cross peaks are obtained in a short-mixing-time (10 ms) TOCSY spectrum; all other vicinal coupling constants are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. For the Down form, the -², ²-³, and ³-² vicinal coupling constants are small, leading to weak TOCSY cross peaks; all other couplings again are in the range 5–12 Hz, and result in medium to strong TOCSY cross peaks. In the case of a Twist conformation, dynamically averaged coupling constants are anticipated. The procedure has been applied to bovine pancreatic trypsin inhibitor and Cucurbita maxima trypsin inhibitor-V, and ring conformations of all prolines in the two proteins have been determined.     

The filtration apparatus of Cladocera: Filter mesh-sizes and their implications on food selectivity

Summary The filtering apparatus of eleven Cladoceran species was studied. The distances between the setulae, which act as filters, were measured. Among adult individuals, they vary from 0.2 m in Diaphanosoma brachyurum to 4.7 m in Sida crystallina. Species can be grouped according to the mesh-sizes, as fine mesh filter-feeders: Diaphanosoma brachyurum, Ceriodaphnia quadrangula, Chydorus sphaericus, Daphnia cucullata and Daphnia magna; medium mesh filter-feeders: Daphnia galeata, D. hyalina. D. pulicaria, Bosmina coregoni, and coarse mesh filter-feeders: Holopedium gibberum and Sida crystallina. In Daphnia hyalina, the distances between setulae increase from 0.3–0.4 m in small juveniles, to 0.8–2.0 m in adults. In Daphnia magna, the mesh-size of the filter does not increase significantly with growth. There is good evidence that the relative abundance of the filter-feeding types varies with the trophic state of the lake. In oligotrophic lakes the coarse mesh filter-feeders usually dominate throughout the year. The seasonal succession of zooplankton species in eutrophic lakes can be interpreted as a succession of feeding types; during winter coarse mesh filter-feeders dominate, while fine mesh filter-feeders are most abundant during summer phytoplankton blooms. Our results support the hypothesis that the species composition of filter-feeding zooplankton is strongly influenced by the amount of suspended bacteria which are available as food only for filter-feeding species with fine meshes.     

Patterns and conformations of commonly occurring supersecondary structures (basic motifs) in protein data bank

Short peptides connecting-helices and-strands have been analyzed in 240 proteins refined at resolutions of 0.25 nm or better. Connecting peptides of lengths between one and five residues have been classified as part of supersecondary motifs of four types:, , , and. Careful consideration has been given to the definition of secondary structures on the basis of hydrogen bonds and main-chain conformational angles. Using five classes of residue conformation—a, b, e, l, t—in the nonregular structure regions of, space, 34 classes of supersecondary motifs occurring at least five times have been identified. Among these 34 classes, 11 classes that occur more than 25 times are commonly occurring supersecondary structure motifs. The patterns and conformations of the 11 commonly occurring supersecondary structure motifs have been characterized, demonstrating that patterns and conformations adopted by supersecondary structure motifs are limited. The results have relevance to structure prediction, comparative modeling, and protein folding.     

Induction of glucoamylase production by non-starchy carbohydrates inAspergillus terreus

Glucoamylase production inAspergillus terreus was induced, in order, by glucose, cellobiose, sorbitol, sucrose, -methyl mannoside and -methyl glucoside. Optimal induction was at 38°C, pH 4.0 and with 8 mg glucose/ml. Cycloheximide at 10 g/ml completely inhibited induction indicatingde novo protein synthesis was involved in induction of glucoamylase.

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Résumé La production de gluco-amylase est induite chezAspergillus terreus, en ordre décroissant, par le glucose, le celloboise, le sorbitol, le sucrose, l'-methylmannoside et l'-methylglucoside. L'induction est optimum à 38°C, pH 4.0 et en présence de 8 mg de glucose parml. La cycloheximide à 10 g par ml inhibe complètement l'induction, ce qui implique use synthèse de protéinede novo dans l'induction de la gluco-amylase.

     

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.     

Essentialistisches und evolution?res Denken in der systematischen Ornithologie des 19. und 20. Jahrhunderts

Zusammenfassung Die Ablösung des essentialistischen (typologischen) Denkens bei Vertretern der systematischen Ornithologie durch das darwinistische (evolutionäre) Populationsdenken um die letzte Jahrhundertwende bildete eine scharfe historische Diskontinuität. Die essentialistische Denkweise, die in Europa während des 19. Jahrhunderts dominierend war, erreichte ihr Ende im Werk vonOtto Kleinschmidt (1870–1954), dem letzten Vertreter derPallas-Schlegel Schule. In den ersten Jahrzehnten dieses Jahrhunderts wurde die evolutionäre Denkweise der Ornithologen der nordamerikanischenBaird-Coues Schule und der europäischenSeebohm-Harter Schule bestimmend. Die weite taxonomische Begrenzung von Arten durch die Ornithologen der essentialistischenPallas-Schlegel Schule und der evolutionären Mikrotaxonomie täuscht Gemeinsamkeiten dieser Schulen vor; im theoretischen Denken ihrer Vertreter bestanden fundamentale Unterschiede. Gegenwärtig ist eine Entwicklung innerhalb der evolutionären Mikrotaxonomie erkennbar, die möglicherweise in naher Zukunft zu einer Trennung in zwei Schulen mit verschiedenen Artkonzepten und unterschiedlicher Artabgrenzung führen wird: (a) Weite Artabgrenzung (mit relativ wenigen polytypischen Arten) und (b) enge Artabgrenzung (mit relativ vielen monotypischen Arten). Dazwischen vermitteln intermediäre Vorschläge. Der wissenschaftliche Naturschutz bezieht sich heute auf evolutionär signifikante Einheiten und ist unabhängig von den verschiedenen systematischen Konzeptionen über die Art.

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Essentialistic and evolutionary thinking in the field of systematic ornithology during the 19^th and 20^th century

Essentialistic thinking dominated systematic ornithology in North America during the first half of the 19^th century and in Europe until the beginning of this century, when evolutionary thinking superseded it marking a sharp historical discontinuity. Evolutionary thinking of the ornithologists of the North AmericanBaird-Coues school and of the EuropeanSeebohm-Hartert school prevailed increasingly from the late 19^th century and early 20^th century onward, respectively. The representatives of the essentialisticPallas-Schlegel school and of evolutionary microtaxonomy delimited species taxa broadly which common procedure concealed the fundamental differences in their theoretical thinking. Within the near future, evolutionary microtaxonomy may split into two different schools supporting different species concepts and a different delimitation of species taxa: (a) Wide species limits resulting in relatively few polytypic species taxa and (b) narrow species limits resulting in numerous monotypic species taxa; intermediate views connect these extremes. Conservation biology refers to evolutionarily significant units and is independent of the different views of systematists as to what constitutes a species.

     

Glycosylation patterns of membrane proteins of the jellyfish Cyanea capillata

Integral and membrane-associated proteins extracted from neuron-enriched perirhopalial tissue of the jellyfish Cyanea capillata were probed with a panel of lectins that recognize sugar epitopes of varying complexity. Of the 13 lectins tested, only concanavalin A, jacalin lectin and tomato lectin stained distinct bands on Western blots, indicating the presence of repeating -1,6-mannoses, terminal Gal--1,6-GalNAc and repeating -1,4-linked GlcNAc, respectively. In whole-mounted perirhopalial tissue, jacalin lectin stained several cell types, including neurons, muscle, cilia and mucus strands. Tomato lectin stained secretory cells intensely, and neurons in a punctate fashion. Concanavalin A stained cytoplasmic epitopes in both ecto-and endodermal cells, and ectodermal secretory cells and the mucus strands emanating from them. With the exception of tomato lectin's sugar epitope, the other sugar epitopes identified in this study are non-complex. This study suggests that while glycosylation of integral and membrane-associated proteins occurs in Cyanea, the sugars post-translationally linked to these proteins tend to be simple.     

The “biological ego”. From Garrod's “chemical individuality” to Burnet's “self”

Starting from the conceptual premises of Garrod, who as long ago as 1902 spoke of chemical individuality, and of Burnet (1949), who recognized as self one's own molecular antigenic structures (as opposed to the antigenic alien: the non- self), the discovery and understanding of HLA antigens and of their extraordinarily individual and differentiated polymorphisms have gained universal recognition. Transplant medicine has now dramatically stressed, within man's knowledge of himself, the characteristic of his biological uniqueness. Today man, having become aware of being a biological antigenic-molecular individuality which is unique and different from that of all of his fellow men (except for monozygotic twins), can therefore easily consider himself a true biological Ego.Abbreviations BMT bone marrow transplantation - GVHD graft versus host disease - HLA human leukocyte antigens - MHC major histocompatibility complex - MLC mixed lymphocyte culture - MLR mixed lymphocyte reaction     

Theβ-subunit of human chorionic gonadotropin containsN-glycosidic trisialo tri- and tri′-antennary carbohydrate chains

TheN-linked carbohydrate chains of the-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz¹H-NMR spectroscopy to beAbbreviations hCG human chorionic gonadotropin - hCG- -subunit - hCG- -subunit - PNGase-F peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52) - endo-F endo--N-acetylglucosaminidase-F (E.C. 3.2.1.96) - SDS sodium dodecyl sulphate - PAGE polyacrylamide gel electrophoresis - CBB coomassie brilliant blue R 250 - GlcNAc N-acetylglucosamine - NeuAc N-acetylneuraminic acid - Man mannose - Gal galactose - Fuc fucose     

The taxonomic significance of the trunk limbs of the chydoridae (Cladocera)

Summary The differences in the structure of the trunk limbs allow to outline three sections of Chydoridae (see table I and fig. 1), coinciding with the sections distinguished according to the structure of the head pores.

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Chydoridae (Cladocera)

Chydoridae (. ), , .

     

Investigations of carotenoids in some faunal elements of the Adriatic Sea. IV-molluscs

The carotenoids in the molluscsClanculus cruciatus, Patella coerulea, Mytilus galloprovincialis, Sepia officinalis andLoligo vulgaris from the Adriatic sea were investigated. Their presence was determined by means of columnar and thin-layer chromatography. The following carotenoids were found inC. cruciatus; mytiloxanthin-like, lutein, lutein ester, zeaxanthin and astaxanthin-like; inP. coerulea: mytiloxanthin-like, lutein, lutein ester, lutein-5,6-epoxide, zeaxanthin and astaxanthin-like; inM. galloprovincialis: -carotene, mytiloxanthin-like, lutein, lutein ester, lutein-5,6-epoxide and zeaxanthin; inS. officinalis: -carotene, lutein, lutein ester, tunaxanthin and zeaxanthin; inL. vulgaris: -carotene, -carotene, -carotene, -cryptoxanthin, isocryptoxanthin, isorenieratene, capsanthin, capsorubin, mutatochrome, triophaxanthin, zeaxanthin, 4-hydroxy--carotene and 4-keto--carotene     

Neutral changes during divergent evolution of hemoglobins

Summary A comparison of the mRNAs for rabbit and human-hemoglobins shows that synonymous changes in codons have accumulated three times as rapidly as nucleotide replacements that produced changes in amino acids. This agrees with predictions based on the so-called neutral theory. In addition, seven codon changes that appear to be single-base changes (according to maximum parsimony) are actually two-base changes. This indicates that the construction of primordial sequences is of limited significance when based on inferences that assume minimum base changes for amino acid replacements.     

Non-reciprocal gonadal dysgenesis in hybrids of the chironomid midge Chironomus thummi II. Gonadal-dysgenesis inducing chromosomes

Females of Chironomus thummi thummi (stock H1) were backcrossed with hybrid males of the cross Ch. thummi thummi (H1) x Ch. thummi piger (E) in order to test which of the four piger chromosomes determine the temperature sensitive non-reciprocal gonadal dysgenesis originally observed in the thummi x piger hybrids. In the backcross rudimentary gonads as well as normal gonads were found irrespective of the chromosome constitution of the individuals. The highest frequency of abnormal gonads (63%) occurred in backcross larvae which received piger chromosomes I and III. Statistical evaluation of the data showed that piger chromosome I is able to induce 44% dysgenic gonads. The gonadal-dysgenesis inducing ability of piger chromosome III is only recognizable if also piger chromosome I is present in the genome. The piger chromosomes II and IV do not have a statistically significant influence on the frequency of abnormal gonads.Arguments are presented for the assumption that the factors determining rudimentary development of the gonads are located distally to those pericentric chromosome regions of the piger chromosomes I and III that in polytene chromosomes show differences in the size of bands in comparison with the homologous thummi regions. The question is discussed whether the rudimentary development of the gonads is caused by gene pairs with an epistatic form of interaction or by the activity of transposable elements.     

Aufbau, Variabilit?t und m?gliche Funktionen des Rufduetts beim ThermometerhuhnLeipoa ocellata

Zusammenfassung Beim ThermometerhuhnLeipoa ocellata tragen die Partner eine Paares ein Rufduett vor. Der Anteil des besteht aus einer Rufreihe, die sich aus einer Folge von 2–7 identischen, zweisilbigen Rufen zusammensetzt. Das trägt einen einzelnen, obertonreichen und langgezogenen Ruf vor. Sowohl der Ruf des als auch die Rufreihe des wird in Serien vorgetragen. Innerhalb einer solcher Ruf- bzw. Rufreihenserie können mehrere Duette auftreten. Die Rufe sind jedoch nicht ausschließlich an das Duett gebunden. Die Variabilität im Aufbau des Duetts äußert sich im Zeitpunkt des Einsatzes des antwortenden Vogels, in der Anzahl der -Rufe während des Duetts und in der Anzahl der Einheiten, aus denen sich der Duettanteil des zusammensetzt. Das beginnt signifikant häufiger als das eine Serie, in der ein oder mehrere Duette vorkommen. Ebenso ist es häufiger der Initiator des ersten in dieser Serie liegenden Duetts. Das Duett dient wohl hauptsächlich zur Festigung des Zusammenhalts zwischen den Paarpartnern. Es erfüllt jedoch von seinen physikalischen Eigenschaften her auch die Bedingungen, die für ein territorial wirksames Signal gelten.

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Structure, variability and possible functions of duetting in the Mallee FowlLeipoa ocellata

Summary In the Australian Mallee Fowl,Leipoa ocellata, both and of a pair are involved in a call duet. The part of the consists of a sequence of 2–7 identical two-syllable calls. The contributes a single long-drawn-out call rich in harmonics. The call of the as well as the call sequence of the are presented in series. Within a series of calls () or call sequences () several duets can occur. The respective vocalizations, however, do not exclusively occur during the duet.The variability in the details of the duet expresses itself in the lag period after which the mate responds, in the number of -calls during the duet, and in the number of calls within the call sequence of the . The begins a series during which one or several duets occur significantly more frequently than the . The circumstances under which duetting occurs indicate that duet calling mainly serves to maintain the pair bond. Moreover, due to its physical characteristics the duet also seems to be suited to serve as a territorial signal.

     

Lectin binding patterns in two cultured endothelial cell types derived from bovine corpus luteum

Epithelial cells of different phenotypes derived from bovine corpus luteum have been studied intensively in our laboratory. In this study, specific lectin binding was examined for cells of type 1 and 3, which were defined as endothelial cells. In order to confirm differences in their glycocalyx at the light microscopic level, five biotinylated lectins were applied to postconfluent cultures which had been fixed with buffered paraformaldehyde or glutaraldehyde. Cells were not permeabilized with any detergent. Lectin binding was localized with a streptavidin-peroxidase complex which was visualized with two different techniques. The DAB technique detected peroxidase histochemically, while the immunogold technique used an anti-peroxidase gold complex together with silver amplification. Neither cell type 1 nor cell type 3 bound a particular lectin selectively, yet each cell type expressed a particular lectin binding pattern. With the DAB technique, diverse lectin binding patterns were seen, probably indicating either outside binding, i.e., a diffuse pattern, a lateral-cell-side pattern and a microvillus-like pattern, or inside binding, i.e., a diffuse pattern, and a granule-like pattern. With the immunogold technique, only outside binding was observed. In addition, the patterns of single cilia or of single circles were detected, the latter roughly representing 3-m-sized binding sites for concanavalin A. When localizing them at the ultrastructural level, single circles corresponded with micron-sized discontinuities of the plasma membrane. Shedding vesicles were detected whose outer embrane was labelled with concanavalin A. Our results confirm the diversity of the two cell types under study. The inside lectin binding may be caused by way of transient plasma membrane openings and related to shedding of right-side out vesicles (ectocytosis).