Immunocytochemical study of Wulzen's cone of the bovine pituitary

Summary The Wulzen's cone of the bovine adenohypophysis presents a variable development and general arrangement. It is joined to the pars intermedia with no intervening connective tissue. It is covered by a single layer of cubical cell epithelium on the side of the hypophysial cleft. Immunofluorescence reveals the presence of different glandular cell types. The most abundant cells are those demonstrated by an anti-oPRL antibody and are either isolated or clustered. Other cells react with anti-hGH, anti-bLH, anti-oLH or anti-hTSH antibodies. Some cells react simultaneously with anti-MSH, anti-^1–24ACTH, anti-^17–39ACTH, anti-LPH and anti-endorphin antibodies. Cell types other than the numerous prolactin cells appear only as isolated elements. We did not observe cells reacting with anti-leu-enkephalin, anti-met-enkephalin or anti-calcitonin antibodies either in the Wulzen's cone or in the pars distalis or pars intermedia.

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Résumé Le cône de Wulzen de l'adénohypophyse bovine présente un développement et une disposition générale variables. Il est accolé à la pars intermedia dont il n'est séparé par aucune cloison conjonctive. Du côté de la fente hypophysaire, il est revêtu par un épithélium simple, cubique. En immunofluorescence, on observe la présence de divers types de cellules glandulaires: les plus abondantes sont des cellules mises en évidence par un anticorps anti-oPRL, isolées ou groupées en amas. D'autres cellules réagissent avec des anticorps anti-hGH, et anti-bLH, anti-oLH ou anti-/mTSH. Quelques cellules réagissent simultanément avec des anticorps anti-MSH, anti-^1–24ACTH, anti-^17–39ACTH, anti-LPH et anti-endorphine. Mises à part les nombreuses cellules à prolactine, les autres types cellulaires apparaissent constamment sous l'aspect d'éléments isolés. Nous n'avons pas observé de cellules réagissant avec des anticorps anti-leu-enképhaline, antiinet-enképhaline ou anti-calcitonine ni dans le cône de Wulzen, ni dans la pars distalis et dans la pars intermedia.

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Abbreviations used in this Article oPRL ovine prolactin - hPRL human prolactin - hGH human growth hormone - bLH bovine luteinizing hormone - oLH ovine luteinizing hormone - pFSH porcine follicle stimulating hormone - hFSH human follicle stimulating hormone - hTSH human thyrotropin - ACTH corticotropin - MSH a melanotropin - MSH melanotropin - LPH lipotropin - PD pars distalis - PI pars intermedia - PN pars nervosa - HC hypophysial cleft     

Immunohistochemical study of the pars tuberalis of the adenohypophysis in the monkey,Macaca irus

Summary The pars tuberalis of the hypophysis in the monkey Macaca irus encompasses the hypophysial stem up to the median eminence. Histologically, it consists of several layers of chromophobic cells. A few PAS¹-positive cells also stainable with Alcian blue (pH 3.0) can be observed among the unstained elements. Using the indirect immunofluorescence antibody technique, scattered immunoreactive cells were revealed with the anti-oLH antibody; these cells did not react with the anti-hFSH antibody. In contrast, the immunoreactions to anti-hGH, anti-hPRL, anti-ACTH, anti-MSH, anti-LPH and anti-endorphin sera were completely negative. Single cells reacting with the anti-hTSH serum were observed at the inferior end of the hypophysial stalk (zona tuberalis), i.e., beyond the pars tuberalis proper. These results are compared with data reported in the literature.

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Résumé La pars tuberalis de l'hypophyse du Singe Macacus irus entoure la tige infundibulaire jusqu'à l'éminence médiane. En techniques histologiques, elle apparaît constituée de plusieurs assises cellulaires d'aspect chromophobe. On y observe quelques cellules PAS-positives réagissant simultanément avec le bleu Alcian (pH3.0). En technique d'immunofluorescence indirecte, des cellules dispersées sont mises en évidence uniquement avec un anticorps anti-oLH; ces cellules ne réagissent pas avec un anticorps anti-hFSH. L'utilisation d'anticorps anti-hGH, anti-hPRL, anti-ACTH, anti-MSH, anti-LPH et antiendorphines ne permet pas de révéler des cellules immunoréactives. Quelques cellules réagissant avec un anticorps anti-hTSH s'observent à la base de la tige hypophysaire (zona tuberalis), c'est-à-dire au-delà de la pars tuberalis proprement dite. Ces résultats sont confrontés à ceux rapportés dans la littérature.

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Abbreviations used in this Article PAS periodic acid Schiff - oLH ovine luteinizing hormone - hFSH human follicle stimulating hormone - hGH human growth hormone - hPRL human prolactin - ACTH corticotropin - MSH melanotropin - LPH lipotropin - hTSH human thyrotropin - BSA and HSA bovine and human serum albumin     

Localisation par immunofluorescence des hormones corticotropes et mélanotropes dans l'hypophyse du rongeur Ellobius lutescens (Th)

Résumé Les réactions d'immunofluorescence induites par l'Ac. anti -(1–24) corticotropine nous ont permis d'identifier les cellules ACTH dans le lobe antérieur de l'hypophyse d'Ellobius lutescens; il s'agit de petites cellules de forme irrégulière, à fins prolongements cytoplasmiques aboutissant à des capillaires et entourant souvent d'autres cellules préhypophysaires; ces mêmes cellules réagissent également, mais d'une façon atténuée, avec l'Ac. anti -MSH. Ce dernier induit une réaction très forte au niveau de toutes les cellules de la pars intermedia, alors que seulement certaines d'entre elles réagissent intensément avec l'Ac. anti -(1–24) corticotropine. L'improbabilité de réactions croisées entre l'Ag. -MSH et l'Ac. -(1–24) corticotropine et vice-versa est discutée. Par ailleurs, seules les cellules de la pars intermedia réagissent avec l'Ac. anti -MSH.

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Immunofluorescent localisation of corticotrophic and melanotrophic hormones in the pituitary gland of the rodent Ellobius lutescens (Th)

Summary Immunofluorescence induced by the antibody to -(1–24) corticotrophin has been used to identify the ACTH cells in the anterior lobe of the pituitary gland in Ellobius lutescens. The cells are small, irregular, with fine cytoplasmic extensions ending near capillaries and often encircling other anterior lobe cell types. They also react, although less strongly, with the antibody to -MSH. The latter antibody induced a marked reaction in all cells of the pars intermedia while only some of them reacted strongly with the antibody to -(1–24) corticotrophin. The unlikeliness of cross-reactions between the antigen -MSH and antibody to -(1–24) corticotrophin is discussed. Furthermore only the cells of the pars intermedia reacted with the antibody to -MSH.

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Abréviations ACTH hormone corticotrope - Ac. anticorps - Ag. antigène - Cell. cellules - C.M.C. fraction inactive de la purification d'ACTH porcine au stade 140 U/mg - C.R.F. fraction inactive de la purification d'ACTH porcine au stade 73 U/mg - Irrég. irrégulières - P.A.S. Periodic Acid-Schiff - P.A. Pars Anterior - P.I. Pars Intermedia     

Detection and partial characterization of two antigenically distinct β-amylases in developing kernels of wheat

A -amylase (EC 3.2.1.2) was identified in the outer pericarp (P) of developing seeds of wheat (Triticum aestivum L.) and compared with the well known -amylase which is synthesized during seed development in the starchy endosperm (E). The enzyme P already exists in the tissues before anthesis and vanishes at the time when E starts to accumulate. The isoelectric-focusing patterns of P and E are very similar. The relative molecular weight (M_r) of P is slightly higher than that of E (66 and 64.5 kDa, respectively). Both P and E exhibit common epitopes in addition to epitopes specific for each of them. The two enzymes were identified in small amounts in the green tissues of the developing seeds (inner pericarp and testa). No antigenic difference was detected between P and the -amylases of roots and leaves.Abbreviations P pericarp -amylase - E endosperm -amylase - IS1 anti--amylase immune serum - IS2 anti- and anti- amylase immune serum - IS3 anti- amylase immune serum - IEF isoelectric focusing - IgG immunoglobulin G The authors thank Dr. P. Ziegler (Universität Bayreuth, FRG) for stimulating discussion and for useful suggestions during the writing of the text. The authors thank Miss C. Mayer for her skillful technical assistance.     

N-andO-alkylation of glycoconjugates and polysaccharides by solid base in dimethyl sulphoxide/alkyl lodide

O-Methylation of simple neutral oligosaccharides is readily accomplished in dimethyl sulphoxide containing solid sodium hydroxide and methyl iodide [Cincanu I, Kerek F (1984) Carbohydr Res 131209-17]. This procedure has been extended to 2-acetamido-2-deoxy sugars and sialic acid-containing oligosaccharides. CompleteO-andN-methylation was in most cases achieved in 15 min. Esterification of carboxylic groups in uronic acids was fast and resulted in concomitant -elimination. The method is also suitable for methylation of glycoproteins and glycosphingolipids. Polysaccharides can also be methylated by this technique. Analysis of the products by gas-liquid chromatography and mass spectrometry showed no degradation products.Abbreviations lacto-N-tetraose LcOse₄, Gal3GlcNAc3Gal4Glc - lacto-N-fucopentaose III III³Fuc-nLcOse₄, Gal4[Fuc3]GlcNAc3Gal4Glc - trihexosylceramide GbOse₃Cer, Gal4Gal4Glc1-1Cer - globoside GbOse₄Cer, GalNAc3Gal4Glc1-1Cer - FAB-MS fas atom bombardment mass spectrometry     

Qualitative and quantitative composition of pigments in Phaeodactylum tricornutum (Bacillariophyceae) stressed by iron

The effect of Fe(III) deficiency on qualitative and quantitative changes in pigment composition in Phaeodactylum tricornutum Bohlin was demonstrated by HPLC and AAS. Maximum content of pigments showed the diatom cells incubated at the optimum iron concentration, i.e., 10 M. The contents of chlorophyll a, chlorophyll c ₁+c ₂, fucoxanthin, diadinoxanthin and ,-carotene were 109.99, 20.16, 40.39, 1.29 and 1.48 fg per cell, respectively. The results obtained showed that Fe(III) affected qualitative and quantitative pigment composition in P. tricornutum. The content of individual pigments, proportions between accompanying pigments and their ratios to chlorophyll a were important indicators of phytoplankton response to iron stress. The strong reduction in ,-carotene content, several times (2–5) increase in diadinoxanthin level as compared to ,-carotene, and high amount of diadinoxanthin in relation to chlorophyll a were observed in algae growing at very low Fe(III) concentrations, 0.001 and 0.01 M. The data suggested that phytoplankton pigments could be a potential physiological marker.     

Infloreszenz- und Blütenentwicklung bei der KugeldistelEchinops exaltatus (Asteraceae)

InEchinops the flowers are surrounded by several scales and initiated in an acropetal and spiral succession on a cone-like inflorescence axis (Figs. 1–6). The floral organs originate in the following sequence: petals—stamens—carpels—pappus. The petals arise from a meristematic rim and therefore are already interconnected when they arise as primordia. This sympetalous zone remains rather inconspicuous for a long period, but eventually, the elongated corolla tube is formed through intercalary growth in a ring zone. Thereby, the stamens are moved upwards and form ledges on the corolla tube (Fig. 34). In the inferior ovary the usual zones of the typical angiospermous gynoecium can be distinguished, namely a synascidiate, symplicate and hemisymplicate zone. The ovule is borne on carpellary tissue.

     

Ibf-1 (Iodine binding factor), a highly variable marker system in the Triticeae

Summary Isoelectric focusing of extracts from the endosperm of mature grains of hexaploid wheat and related species was used to study the genetic control of Iodine binding factor (IBF). Ten IBF bands were present in Chinese Spring (CS) and analysis of the nullisomictetrasomic and ditelosomic lines of CS showed nine of them to be controlled by genes on the long arms of the homoeologous group 5 chromosomes. Five alleles were detected at Ibf-A1 locus, four at Ibf-B1 and four at Ibf-D1 among a sample of 46 wheat genotypes. Homoeoloci were found on chromosome 5R of Secale cereale, 5E of Agropyron elongatum, 5U of Aegilops umbellulata, 5Agⁱ of Agropyron intermedium, 5S¹ and 4S¹ of Aegilops sharonensis and 4H of Hordeum vulgare.     

β-Adrenoceptors in the Tree Shrew Brain. I. Distribution and Characterization of [125I]Iodocyanopindolol Binding Sites

1. The number and distribution pattern of -adrenergic receptors in the brain have been reported to be species specific. The aim of the present study was to describe binding of the -adrenoceptor ligand [¹²⁵I]iodocyanopindolol in the brain of the tree shrew (Tupaia belangeri), a species which provides an appropriate model for studies of psychosocial stress and its consequences on central nervous processes.2. ¹²⁵I-Iodocyanopindolol (¹²⁵ICYP) labeling revealed a high degree of nonspecific binding, which was due mainly to interactions of this ligand with serotonin binding sites. For a quantitative evaluation of ₁- and ₂-adrenoceptors, serotonin binding sites had to be blocked by 100 M 5HT.3. Binding of the radioligand to ₁- and ₂-adrenoceptors was characterized using the ₁-specific antagonist CGP20712A and the ₂-specific antagonist ICI118.551. ₁-adrenoceptor binding is present in the whole brain, revealing low receptor numbers in most brain regions (up to 1.5 to 2.7 fmol/mg). A slight enrichment was observed in cortical areas (lateral orbital cortex: 4.0±0.7 fmol/mg) and in the cerebellar molecular layer (8.7±1.0 fmol/mg).4. Competition experiments demonstrated high- and low-affinity binding sites with considerable variations in K _i values for CGP20712A, showing that various affinity states of ₁-adrenoceptors are present in the brain (K _i: 0.61 nM to 67.1 M). In the hippocampus, only low-affinity ₁-adrenoceptors were detected (K _i: 1.3±0.2 M). Since it is known that ¹²⁵ICYP labels not only membrane bound but also internalized -adrenoceptors, it can be assumed that the large population of the low-affinity sites represents internalized receptors which may be abundant due to a high sequestration rate.5. High numbers of ₂-adrenoceptors are present in only a few brain structures of tree shrews (external layer of the olfactory bulb, 15.8±2.0 fmol/mg; claustrum, 19.3±1.5 fmol/mg; anteroventral thalamic nucleus, 19.4±1.5 fmol/mg; cerebellar molecular layer, 55.0±4.3 fmol/mg). Also for this class of -adrenoceptors, high- and low-affinity binding sites for the ₂-selective antagonist ICI118.551 were observed, indicating that ¹²⁵ICYP labels membrane bound and internalized ₂-adrenoceptors. Only in the cerebellar molecular layer was a high percentage of high-affinity ₂-adrenoceptors detected (K _i for ICI118.551 was 1.8±0.3 nM for 90% of the receptors).6. In conclusion, ₁- and ₂-adrenoceptor binding can be localized and quantified by in vitro receptor autoradiography in the brains of tree shrews when serotonergic binding sites are blocked. Modulatory effects of long-term psychosocial conflict on the central nervous -adrenoceptor system in male tree shrews are described in the following paper.     

Variability in the 11S globulin fraction of seed storage proteins ofHelianthus (Asteraceae)

The seed storage globulins from sixHelianthus and four hybrids were studied using mono and bidimensional gel SDS electrophoresis (+ 2 mercaptoethanol). The polypeptide composition of each subunit was determined. Different pairs are specifically expressed according to the species studied. Three typical patterns were discriminated. All the studied species exhibit five subunits: two of them are expressed in all the species (₁₁ and ₂₂). The subunit corresponding to the ₁₁ pair is present inH. petiolaris and in the three populations ofH. annuus studied. The _2b2 pair is common toH. annuus andH. argophyllus. H. petiolaris presents two specific _2a2 and ₄₄ pairs andH. annuus a specific ₃₃ pair. InH. argophyllus _{11 33} or ₄₄ are never observed but are replaced by ₁₃ and ₃₁ pairs. Some globulins, poorly represented, are of forms but present chains of higher molecular weights (in the range 54–56 kDa). Expressing variations in the banding patterns between these species by the use of a similarity index reveals complete identity between the three populations ofH. annuus. Identity between the twoH. petiolaris studied is also observed.H. annuus andH. argophyllus appear to be closer to each other thanH. petiolaris concerning the seed storage globulins.     

Steroid Polyols from the Far Eastern Starfish Henricia sanguinolenta and H. leviuscula leviuscula

Two new asterosaponins, (20R)-3-O--D-(2-O-methylxylopyranosyl)-24-propylcholest-4-ene-3,6,8,15,16,29-hexaol (sanguinoside A) and (20R,24S)-3-O--D-(2,3,4-tri-O-methylxylopyranosyl)-5-cholestane-3,4,6,8,15,24-hexaol (sanguinoside B), were isolated from two species of Pacific Far Eastern Starfish Henricia sanguinolenta and H. leviuscula leviuscula, collected in the Sea of Okhotsk. Both glycosides contain aglycones with pentahydroxysteroid nuclei of similar structures, which are substituted at the 3-hydroxy group with differently methylated -D-xylosyl residues. Sanguinoside A has an unusual structure of its aglycone side chain, whereas sanguinoside B has a unique permethylated carbohydrate chain. In addition, laevisculoside G, a known glycoside, was identified in the H. leviuscula starfish. The structures of the isolated glycosides were established by interpreting their spectral data and by comparing their spectral characteristics with those of known compounds.     

Expression of the β-subunit of β-conglycinin in seeds of transgenic plants

Soybean (Glycine max (L.) Merr.) seeds contain the storage protein -conglycinin, encoded by a multigene family. -Conglycinin consists of three subunits; , , and . A genomic clone for a -subunit of -conglycinin has been characterized by restriction-enzyme mapping and hybrid selected in-vitro translation followed by immunoprecipitation. In order to determine the developmental regulation of this -subunit gene, its expression was studied in seeds of transgenic petunia (Petunia hybrida) and tobacco (Nicotiana tabacum L.) plants. The -subunit expressed in seeds of petunia and tobacco was recognized by anti--conglycinin serum at a relative molecular mass of 53 000, equivalent to that of the native protein. Separation of the petunia-seed proteins by isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis showed that multiple isoelectric forms of the -subunit were produced. There was approximately a twofold variation in the accumulation of the -subunit protein in the mature seeds of transgenic petunia plants, each containing a single -subunit gene. However, the level of protein accumulation in mature seeds and the amount of -subunit mRNA in developing seeds was not correlated. Accumulation of the -subunit protein in transgenic seeds was less than the -subunit protein that accumulated in transgenic petunia seeds containing a single -subunit gene and less than the amount of the -subunit in mature soybean seeds which contain 8–13 -subunit genes. In transgenic tobacco plants, the accumulation of the -subunit protein in seeds was generally well correlated with the number of genes that were incorporated in the different transformants.Abbreviations kb kilobase - kDa kilodalton - M_r relative molecular mass - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis     

Presence of carotenoids in Gordius aquaticus (Nematomorpha,Nemathelminthes) specimens from deep wells

By means of columnar and thin-layer chromatography, the presence of carotenoids in Gordius aquaticus L. (Nematomorpha, Nemathelminthes) from deep wells was studied.The investigations revealed the presence of the following carotenoids: -carotene, mutatochrome, -cryptoxanthin, ,-carotene epoxide,lutein, zeaxanthin and astaxanthin ester.     

Purification and immunocytochemical localization of kafirins inSorghum bicolor (L. Moench) endosperm

Summary Kafirins are the storage proteins of sorghum and are found in protein bodies in the seed endosperm. They have been classified as -, -, and -kafirins according to differences in molecular weight, solubility, and structure. The kafirins were purified, amino acid composition was determined, and immunolocalization methods were used to determine the organization of the protein bodies and distribution of kafirins throughout the endosperm. All three groups of kafirins were low in lysine. -Kafirins and -kafirins were relatively high in cysteine, and -kafirins were relatively high in methionine. Transmission electron microscopy showed that protein bodies in the peripheral endosperm were spheroid with concentric rings and few darkly stained inclusions. In contrast, protein bodies of the central endosperm were irregularly shaped with a higher proportion of darkly stained material. The light staining regions of the protein bodies are composed primarily of -kafirins with minor portions of - and -kafirins. The dark staining regions, however, are composed primarily of - and -kafirins. Immunoelectron microscopy showed that protein bodies in the peripheral endosperm contain predominantly a-kafirin with minor amounts of - and -kafirin. Central endosperm protein bodies are also predominantly -kafirin, but have a higher proportion of -kafirin and -kafirin than the peripheral endosperm protein bodies.Abbreviations GAR-HRP Goat anti-rabbit horseradish peroxidase - IgG immunoglobulin G - 2-ME 2-mercaptoethanol - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis - TBS Tris buffer saline - TBS-T Tris buffer saline with Tween - TBS-T-B Tris buffer saline with Tween and bovine serum albumin - TCA trichloroacetic acid - UV ultraviolet     

Comparative cellular localization of corticotropin and melanotropin in lerot adenohypophysis (Eliomys quercinus)

Summary Corticotropin and melanotropin producing cells were localized in the adenohypophysis of normal Lerots by using antibodies against synthetic corticotropins (anti 1–24 ACTH, anti 17–39 ACTH, anti 25–39 ACTH), and melanotropins (anti MSH, anti MSH). All the anticorticotropin sera stained the same cells both in the anterior lobe and in the intermediate lobe. The anti MSH serum only stained a few cells, exclusively located in the intermediate lobe. These MSH cells were not stained with anticorticotropin antibodies. The anti MSH serum revealed all the cells stained with anticorticotropin and anti MSH sera. Absorption tests showed that the 4–10 heptapeptide common to ACTH and MSH, is not responsible for the immunohistochemical staining. The staining of only some corticotrophs with the anti 4–10 ACTH serum might indicate the presence in these cells of a peptide with an accessible 4–10 site. These results are discussedWe thank A. Pillez for technical assistance (C.N.R.S.). This work was supported by a grant from U.E.R. III Lille 1976Attaché de Recherche INSERM     

The tryptic peptide composition of the beta chains of hemoglobins A and B of the domestic cat (Felis catus)

Summary The tryptic peptides from the ^A and ^B chains of cat hemoglobins A and B have been isolated and the amino acid compositions determined. Differences between the two chains were found in two peptides,T-1 (GlySer) andT-14 (AsnSer and LysArg). The GlySer and LysArg substitutions are placed at-1 and-144 respectively from earlier work, and the third substitution, AsnSer at-139 is suggested from this work. In addition, the presence of a blocked amino terminus in ^B has been confirmed. Tentative sequences constructed by homology with known-chain structures suggest the occurrence of substitutions at _{1 1} contacts in ^A and ^B that may be functionally significant. There are at least 18 differences in amino acid composition between cat ^A and dog-chains and 22 differences between cat ^A and normal adult human-chains.     

Microbial transformation of ginsenoside Rb1 by Rhizopus stolonifer and Curvularia lunata

Of 49 microbial strains screened for their capabilities to transform ginsenoside Rb₁, Rhizopus stolonifer and Curvularia lunata produced four key metabolites: 3-O-[-d-glucopyranosyl-(1,2)--d-glucopyranosyl]- 20-O-[-d-glucopyranosyl]-3,12, 20(S)-trihydroxydammar-24-ene (1), 3-O-[-d-glucopyranosyl-(1,2)--d- glucopyranosyl]-20-O-[-d-glucopyranosyl]-3,12, 20(S)-trihydroxydammar-24-ol (2), 3-O-[-d-gluco- pyranosyl-(1,2)--d-glucopyranosyl]-3, 12, 20(S)-trihydroxydammar-24-ene (3), and 3-O--d-glucopyranosyl-3, 12, 20(S)-trihydroxydammar-24-ene (4), identified by TOF-MS, ¹H- and ¹³C-NMR spectral data. Metabolites 1, 3 and 4 were from the incubation with R. stolonifer, and 1 and 2 from the incubation with C. lunata. Compound 2 was identified as a new compound.     

A quantitative method for separating reaction components of CMP-sialic acid: Lactosylceramide sialyltransferase using Sep Pak C18 cartridges

A rapid procedure is described for the separation of CMP-sialic acid:lactosylceramide sialyltransferase reaction components using Sep Pak C₁₈ cartridges. The quantitative separation of the more polar nucleotide sugar, CMP-sialic acid, and its free acid from the less polar G_M3-ganglioside is simple and rapid relative to previously described methods. Recovery of G_M3 is optimized by the addition of phosphatidylcholine to the reaction mixture prior to the chromatographic step. Using rat liver Golgi membranes as a source of CMP-sialic acid: lactosylceramide sialyltransferase activity (G_M3 synthase; ST-1), the transfer of [¹⁴C] sialic acid from CMP-[¹⁴C] sialic acid to lactosylceramide can be quantified by this assay. The procedure is reliable and may be applicable to the isolation of ganglioside products in otherin vitro glycosyltransferase assays.Abbreviations G_M3 G_M3-ganglioside - II³NeuAc-LacCer NeuAc2-3Gal1-4Glc1-1Cer - G_D1a G_D1a-ganglioside, IV³NeuAc, II³NeuAc-GgOse₄Cer, NeuAc2-3Gal1-3GalNac1-4(NeuAc2-3)Gal1-4Glc1-1Cer - G_D3 G_D3-ganglioside, II³(NeuAc)₂LacCer, NeuAc2-8NeuAc2-3Gal1-4Glc1-1Cer - GgOse₄Cer asialo-G_M1 Gal1-3GalNAc1-4Gal1-4Glc1-1Cer - FucG_MI fucosyl-G_MI-ganglioside, Fuc1-2Gal1-3GalNAc1-4Gal1-4 Glc1-1Cer - ST-1 G_M3 synthase, CMP-sialic acid:lactosylceramide sialyltransferase - LacCer lactosylceramide, Gal1-4Glc1-1Cer - CMP-NeuAc cytidine 5-monophospho-N-acetylneuraminic acid - PC phosphatidylcholine - PMSF phenylmethylsulfonyl fluoride     

Production of a novel syrup containing neofructo-oligosaccharides by the cells of Penicillium citrinum

A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.     

Carbohydrates of influenza virus. Structure of the oligosaccharides linked to asparagines 406 and 478 in the hemagglutinin of fowl plague virus,strain dutch

Fowl plague virus, strain Dutch, was metabolically labeled withd-[2-³H]mannose, or withd-[6-³H]glucosamine, and the small subunit (HA₂; 0.8 mg in total) of the viral hemagglutinin was isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis. After proteolytic digestion, the radioactive oligosaccharides were sequentially liberated from the glycopeptides by treatment with different endo--N-acetylglucosaminidases and with peptide:N-glycosidase or, finally, by hydrazinolysis. In this manner, four groups of glycans could be obtained by consecutive gel filtrations and were subfractionated by HPLC. The structures of the individual oligosaccharides were analyzed by micromethylation, by acetolysis or by digestion with exoglycosidases. The major species amongst the high mannose glycans at Ans-406 of the viral glycopolypeptide were found to be Man1-2Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNac1-4GlcNAc and Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNAc1-4GlcNAc, while the complex glycans at Asn-478 are predominantly GlcNAc1-2Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc (lacking, in part, one of the outerN-acetylglucosamine residues) and GlcNAc1-2Man1-3(Gal1-4GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc.Abbreviation BSA bovine serum albumin - endo D (F,H) endo--N-acetyl-d-glucosaminidase D (F,H) - HA hemagglutinin (HA₁, large subunit of HA - HA₂ small subunit - FPV fowl plague virus - PNGase F peptide:N-glycosidase F - SDS sodium dodecylsulfate