Identification of phosphate granules occurring in seedling tissue of two palm species (Phoenix dactylifera and Washingtonia filifera)

Haustoria of two palm species, Phoenix dactylifera L. (date) and Washingtonia filifera (Lindl.) Wendl were carefully dissected from seeds, and the ultrastructural characteristics of the large, electron-opaque granules present in the cells of this tissue were compared using standard aldehyde and OsO₄ fixations. In Washingtonia, the granules were smaller than those in date and were more variable in size and presence of non-opaque inclusions. All granules appeared to be membrane bounded although they often filled the bounded space. No protein, lipid, carbohydrate or tannins were found in the granules by standard staining procedures. The granules stained positively with two different metallic-phosphate stains which contained either bismuth or lead. Energy dispersive X-ray microprobe analysis, done on aldehyde-fixed granules and those stained with both phosphate stains, confirmed the fact that phosphorus and calcium were present in the granules. The granules also bound the metallic stains as expected. All procedures consistently confirmed the presence of phosphate in the granules. The data are most consistent with the hypothesis that the granules are composed of polyphosphate.Abbreviations and symbols EDAX energy-dispersive X-ray microanalysis - K K shell peak - K K shell peak - L L shell peak - L L shell peak - M M shell peak     

L cells expressing DQ molecules of the DR3 and DR4 haplotypes: reactivity patterns with mAbs

cDNAs coding for the HLA class II DR and DQ and chains of the diabetogenic haplotypes DR3 and DR4 were introduced into a mammalian expression vector and transfected into L-cell mouse fibroblasts to produce cells expressing individual human class II molecules. Stable L transfectants were generated expressing each of the DR or DQ isotypes of the cis-encoded and chains of the DR3 or DR4 haplotypes, as well as the trans-encoded and chains of the DQ molecules of the two haplotypes. However, isotype mismatched combinations (DR/DQ or DQ/DR) did not result in any stable transfectants. The stable DQ L-cell transfectants obtained, along with homozygous B-cell lines expressing the DQ2 and DQ8 specificities, were tested against a large panel of twentyone anti-HLA class II monoclonal antibodies (mAbs). Their unusual reactivity patterns are described including the failure of most pan-DQ mAbs to react with all DQ expressing L-cell transfectants. Interestingly, some mAbs react with certain heterodimers expressed on B-LCL but fail to recognize the same heterodimers expressed on the transfectants. This is suggestive of minor structural modifications that class II molecules undergo depending on the cells they are expressed on.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession number U07848. The name DQB1 ^* 0202 was officially assigned by the WHO Nomenclature Committee in April 1994. This follows the agreed policy that, subject to the conditions stated in the most recent Nomenclature Report (Bodmer et al. 1992), names will be assigned to new sequences as they are identified. Lists of such new names will be published in the following WHO Nomenclature Report     

Arabidopsis thaliana ‘extra-large GTP-binding protein’ (AtXLG1): a new class of G-protein

Heterotrimeric GTP-binding proteins, composed of , , and subunits, are involved in signal transduction pathways in animal and plant systems. In plants, physiological analyses implicate heterotrimeric G-proteins in ion channel regulation, light signaling, and hormone and pathogen responses. However, only one class of plant G genes has been identified to date. We have cloned a novel gene, Arabidopsis thaliana extra-large GTP-binding protein (AtXLG1). AtXLG1 appears to be a member of a small gene family and is transcribed in all tissues assayed: roots, leaves, stems, flowers, and fruits. The conceptually translated protein from AtXLG1 is 99 kDa, twice as large as typical G proteins. The carboxy-terminal half of the AtXLG1 protein has significant homology to animal and plant G proteins. This region includes a GTP-binding domain, a predicted helical domain, and an aspartate/glutamate-rich loop, which are characteristics of G's. Despite the absence of some of the amino acids implicated in GTP binding and hydrolysis by crystallographic and mutational analyses of mammalian G's, recombinant AtXLGl binds GTP with specificity. The amino-terminal region of AtXLGl contains domains homologous to the bacterial TonB-box, which is involved in energy transduction between the inner and outer bacterial membranes, and to zinc-finger proteins. Given the unique structure of AtXLG1, it will be of interest to uncover its physiological functions.     

Stochastic prey arrivals and crab spider giving-up times: simulations of spider performance using two simple “rules of thumb”

Summary We compared the patch-choice performances of an ambush predator, the crab spider Misumena vatia (Thomisidae) hunting on common milkweed Asclepias syriaca (Asclepiadaceae) umbles, with two stochastic rule-of-thumb simulation models: one that employed a threshold giving-up time and one that assumed a fixed probability of moving. Adult female Misumena were placed on milkweed plants with three umbels, each with markedly different numbers of flower-seeking prey. Using a variety of visitation regimes derived from observed visitation patterns of insect prey, we found that decreases in among-umbel variance in visitation rates or increases in overall mean visitation rates reduced the clarity of the optimum (the difference in the yield obtained as foraging behavior changes), both locally and globally. Yield profiles from both models were extremely flat or jagged over a wide range of prey visitation regimes; thus, differences between optimal and next-best strategies differed only modestly over large parts of the foraging landscape. Although optimal yields from fixed probability simulations were one-third to one-half those obtained from threshold simulations, spiders appear to depart umbels in accordance with the fixed probability rule.     

Degradation of barley β-glucan by endoglucanase C of Clostridium thermocellum

Summary Endoglucanase C encoded by the celC gene of Clostridium thermocellum has been purified to homogeneity from a recombinant Escherichia coli strain. It was found that this enzyme is highly efficient in degrading glucans with alternating -1,4- and -1,3-linkages but lacks activity on unmodified cellulosic substrates. The properties of endoglucanase C were compared to those of Bacillus subtilis -glucanase, an enzyme used in the brewing industry for -glucan degradation. Both enzyme cause a rapid decrease of the viscosity of barley -glucan as a result of internal chain cleavage. Endoglucanase C hydrolyses non-specifically -1,3- and -1,4-bonds adjacent to unsubstituted or 4-O-substituted cellobiose units. Due to its lower pH optimum and increased thermostability endoglucanase C compares favourably with B. subtilis -glucanase and seems suitable for use in the mashing process of beer brewing.     

Oligosaccharide Specificity of the Fucolectin from the Bark of Laburnum (Laburnum anagyroides)

A comparative study of fine carbohydrate specificity of the lectin from the bark of laburnum Laburnum anagyroides (LABA) and the fucolectin from asparagus pea Tetragonolobus purpureus (TPA) was performed using inhibition of agglutination of the complex formed by H-active neoglycoprotein and nanoparticles of colloidal gold. Both lectins bound most strongly the H type 2 oligosaccharides comprising O-glycans; however, LABA was almost unable to discriminate between them. LABA bound more weakly the H type 6 trisaccharide (Fuc1-2Gal1-4Glc) and difucosyllactose (Fuc1-2Gal1-4[Fuc1-3]Glc), a glucoanalogue of the Le^y antigen, and, even more weakly, the Le^a pentasaccharide lacto-N-fucopentaose II (Gall-3[Fucl-4]GlcNAcl-3Gall-4Glc). However, LABA did not bind the antigens Le^b, Le^c, and Le^d, very poorly interacted with the terminal Le^x, and somewhat more strongly bound the internal Le^x. The lectin also had a hydrophobic binding site. Both lectins exhibited a cluster effect with polymeric ligands (neoglycoproteins).     

A re-examination of the Na+-independent binding of [3H]β-alanine to rat brain stem-spinal cord

The Na⁺-independent binding of [³H]-alanine to rat brain stem plus spinal cord was reinvestigated, in order to study in more detail the characteristics of previously described -alanine binding processes. Binding was absent when amino acid-free postnuclear supernatants or crude synaptic membranes were used. Experiments performed with several other Na⁺-free preparations showed a sole binding component, irrespective of the preparation used. Biochemical characterization of this Na⁺-independent binding, using frozen/thawed/washed synaptosomal-mitochodrial fractions, showed that binding reached a plateau between 7 min and 13 min, increasing thereafter. Binding was linear with fraction protein over a range of 200–415 g/ml incubation medium. Binding was completely inhibited by glycine, alanine, -aminobutyric acid, -aminoisobutyric acid, hypotaurine and strychnine, and to a lesser extent by 2,2-dimethyl--alanine, brucine and gelsemine. It was insensitive to taurine, -aminobutyric acid (GABA), 2-guanidinoethanesulfonic acid (GES), carnosine, and bicuculline methiodide. Binding was reversible, saturable (K _D 20 M), and heat sensitive.     

Average phase difference theory and 1∶1 phase entrainment in interlimb coordination

The dynamics of coupled biological oscillators can be modeled by averaging the effects of coupling over each oscillatory cycle so that the coupling depends on the phase difference between the two oscillators and not on their specific states. Average phase difference theory claims that mode locking phenomena can be predicted by the average effects of the coupling influences. As a starting point for both empirical and theoretical investigations, Rand et al. (1988) have proposed d/dt= — K sin ), with phase-locked solutions =arcsin( /K), where is the difference between the uncoupled frequencies and K is the coupling strength. Phase-locking was evaluated in three experiments using an interlimb coordination paradigm in which a person oscillates hand-held pendulums. was controlled through length differences in the left and right pendulums. The coupled frequency _c was varied by a metronome, and scaled to the eigenfrequency _v of the coupled system K was assumed to vary inversely with _c. The results indicate that: (1) and K contribute multiplicatively to (2) =0 or = regardless of K when =0; (3) 0 or regardless of when K is large (relative to ); (4) results (1) to (3) hold identically for both in phase and antiphase coordination. The results also indicate that the relevant frequency is _c/_v rather than _c. Discussion high-lighted the significance of confirming =arcsin(/K) for more general treatments of phase-locking, such as circle map dynamics, and for the 11 phase-entrainment which characterizes biological movement systems.     

Zur Lokalisierung von Zellkontakten zwischenCuscuta odorata und verschiedenen höheren Wirtspflanzen

Zusammenfassung Es wird eine Methode der gezielten Untersuchung von parasitierendenCuscuta-Hyphen innerhalb des Wirtsgewebes (Pelargonium zonale, Primula obconica) beschrieben. Das Arbeiten mit isolierten Leitbündeln (Primula) sowie eine verfeinerte Zielpräparation der fixierten und eingebetteten Gewebe vor der Ultramikrotomie führt bereits mit lichtoptischen Methoden zu einer zellularen Lokalisierung der Hyphen im Wirtsgewebe. Diese wiederum ermöglicht eine genaue Beurteilung des Differenzierungsstadiums der Parasitenzellen vor der nachfolgenden elektronenoptischen Beobachtung. Erste Ergebnisse über den Feinbau derCuscuta-Hyphen werden an drei Beispielen mitgeteilt: 1. intrazellular wachsende Suchhyphe, 2. interzellulare Suchhyphe und 3. Anschlußhyphe an der Wirtssiebröhre. Die Untersuchungen wurden in den Jahren 1965–1966 im Botanischen Institut der Universität Bonn durchgeführt. Mein herzlicher Dank gilt meinem verehrten Lehrer Herrn Prof. Dr. W.Schumacher. Danken möchte ich ganz besonders Herrn Doz. Dr. R.Kollmann für seine Einführung in das Arbeitsgebiet sowie für alle anregenden Diskussionen.

---

Localization of cellcontacts betweenCuscuta odorata and different higher hostplants

Summary A new method for localized research of parasitic dodder hyphae in host tissue (Pelargonium zonale, Primula obconica) is described. Working with isolated vascular bundles (Primula) and the use of a refined, aimed preparation enable a cellular localization of theCuscuta hyphae in the host tissue already by the lightmicroscope. This facilitates an exact analyse of the stage of differentiation of the the parasitic cells previous to the ultramicrotomy and electronmicroscopic observation. First findings on the ultra-structure ofCuscuta hyphae are reported in three examples: 1. intracellular growing searching hypha, 2. intercellular searching hypha, and 3. contact hypha to the sieve tube of the host plant.

---

---

Teilabdruck einer Dissertation der Mathem.-Naturwiss. Fakultät der Universität Bonn. Das vollständige Exemplar kann an obiger Stelle eingesehen werden.     

A cytochemical method for visualising blood vessels selectively in semithin LR white sections

Traditional cytochemical techniques for illustrating blood vessels are usually unsuitable for use in high resolution, 0.35m-thick LR White sections due to the thinness of the tissue and, if collagenous, high background staining. We report that a modification of the periodic acid-thiocarbohydrazide-silver proteinate-silver enhancement method (PATCH-SP-SE) selectively visualises, with low background, even the smallest blood vessels in semithin LR White sections of highly collagenous tissues such as human parietal peritoneum.     

Evolutionary origin of the class A and class C β-lactamases

Summary The protein sequences of 18 class A -lactamases and 2 class C -lactamases were analyzed to produce a rooted phylogenetic tree using the DD peptidase of Streptomyces R61 as an outgroup. This tree supports the penicillin-binding proteins as the most likely candidate for the ancestoral origin of the class A and class C -lactamases, these proteins diverging from a common evolutionary origin close to the DD peptidase. The actinomycetes are clearly shown as the origin of the class A -lactamases found in other non-actinomycete species. The tree also divides the -lactamases from the Streptomyces into two subgroups. One subgroup is closer to the DD peptidase root. The other Streptomyces subgroup shares a common branch point with the rest of the class A -lactamases, showing this subgroup as the origin of the non-actinomycete class A -lactamases. The non-actinomycete class A -lactamase phylogenetic tree suggests a spread of these -lactamases by horizontal transfer from the Streptomyces into the non-actinomycete gram-positive bacteria and thence into the gram-negative bacteria. The phylogenetic tree of the Streptomyces class A -lactamases supports the possibility that horizontal transfer of class A -lactamases occurred within the Streptomyces.     

Hydrolysis of linalyl acetate and α-terpinyl acetate by yeasts

Of 104 yeasts tested for the hydrolysis of rac-linalyl acetate and rac--terpinyl acetate twelve hydrolysed rac-linalyl acetate. Five of these belonged to five different species of Geotrichum. The most promising strain, Geotrichum capitatum CBS 0572.82, yielded (S)-linalool in 56% enantiomeric excess after 6h (E value 5, c = 32%). Only one unclassified yeast isolate hydrolyzed rac--terpinyl acetate with a slight preference for the S-enantiomer (E value 2). © Rapid Science Ltd. 1998     

Some observations on cyclodextrin-mediated boosting of secreted amylolytic enzymes byLactobacillus amylovorus

The amounts of a 160-kDa amylase and a 140-kDa -amylase (A. Burgess-Cassler and S. H. Imam, Curr. Microbiol. 23:207–213, 1991) secreted into culture medium by the starchutilizingLactobacillus amylovorus were enhanced by the use of cyclodextrin (CD) as the carbon source. The levels of total extracellular -amylase obtained with glucose as the carbon source could be boosted severalfold by use of CD. The best enhancer was -CD, and the rank order of best to least effective was -CD>-CD=-CD>glucose.Another amylase, a 65-kDa -amylase, which degraded para-nitrophenyl-(1,4)-d-glucopyranoside, was also detected in this study. The most effective enhancer in this case was -CD, and the rank order was -CD>-CD>-CD glucose. Despite its ability to degradep-nitrophenylated glucose, this enzyme did not convert maltose to glucose. It showed a cleared zone on starch zymograms and did degrade short maltodextrins to maltose. Neither this new -amylase nor the 140-kDa -amylase exhibited any detectable ring-decyclizing (cyclodextrinase) activity against -or -CD.Other extracellular amylases (not characterized here) appeared to be similarly enhanced by CDs. Although the precise mechanism by which this effect is accomplished remains undefined, CDs can be useful inducing agents, boosting the expression and/or secretion of otherwise low-level enzymes, either as additives to growth media or as sole carbon source.     

On the sufficiency of the velocity field for perception of heading

All models of self-motion from optical flow assume the instantaneous velocity field as input. We tested this assumption for human observers using random-dot displays that simulated translational and circular paths of movement by manipulating the lifetime and displacement of individual dots. For translational movement, observers were equally accurate in judging direction of heading from a velocity field with a two-frame dot life and a direction field in which the magnitudes of displacement were randomized while the radial pattern of directions was preserved, but at chance with a speed field in which the directions were randomized, preserving only magnitude. Accuracy declined with increasing noise in vector directions, but remained below 2.6° with a 90° noise envelope. Thus, the visual system uses the radial morphology of vector directions to determine translational heading and can tolerate large amounts of noise in this pattern. For circular movement, observers were equally accurate with a 2-frame velocity field, 3-frame acceleration displays, and 2-frame and 3-frame direction fields, consistent with the use of the pattern of vector directions to locate the center of rotation. The results indicate that successive independent velocity fields are sufficient for perception of translational and circular heading.     

Multiple sex-chromosomes in the common Indian Krait,Bungarus caeruleus Schneider

The highly poisonous common Indian Krait, Bungarus caeruleus of the family Elapidae has 2n =44 and 2n =43. There is a sharp difference in size between macro and microchromosomes. The number of macrochromosomes is 24 in males and 23 in females whereas there are 20 microchromosomes in both the sexes. The difference in number of macro-chromosomes in the two sexes is explainable on the basis of translocation of a macro-autosome to the W chromosome resulting in a multiple sex-chromosome constitution of Z₁Z₁Z₂Z₂/Z₁Z₂W-types. Autoradiographic studies using H³-TdR show that the W is late replicating except for the translocated part which finishes its replication along with the macroautosomes. This is the first example of a multiple sex-chromosome complex in a vertebrate with female heterogamety.     

In vivo fucosylation of lacto-N-neotetraose and lacto-N-neohexaose by heterologous expression of Helicobacter pylori alpha-1,3 fucosyltransferase in engineered Escherichia coli

We report here the in vivo production of type 2 fucosylated-N-acetyllactosamine oligosaccharides in Escherichia coli. Lacto-N-neofucopentaose Gal1-4GlcNAc1-3Gal1-4(Fuc1-3)Glc, lacto-N-neodifucohexaose Gal1-4(Fuc1-3)Glc-NAc1-3Gal1-4(Fuc1-3)Glc, and lacto-N-neodifucooctaose Gal1-4GlcNAc1-3Gal1-4(Fuc1-3)GlcNAc1-3Gal1-4(Fuc1-3)Glc were produced from lactose added in the culture medium. Two of them carry the Lewis X human antigen. High cell density cultivation allowed obtaining several grams of fucosylated oligosaccharides per liter of culture. The fucosylation reaction was catalyzed by an -1,3 fucosyltransferase of Helicobacter pylori overexpressed in E. coli with the genes lgtAB of N. meningitidis. The strain was genetically engineered in order to provide GDP-fucose to the system, by genomic inactivation of gene wcaJ involved in colanic acid synthesis and overexpression of RcsA, positive regulator of the colanic acid operon.To prevent fucosylation at the glucosyl residue, lactulose Gal1-4Fru was assayed in replacement of lactose. Lactulose-derived oligosaccharides carrying fucose were synthesized and characterized. Fucosylation of the fructosyl residue was observed, indicating a poor acceptor specificity of the fucosyltransferase of H. pylori.     

Concentration/response effect of 2,2′, 4,4′, 5,5′-hexabromobiphenyl on cell-cell communication in vitro: assessment by fluorescence redistribution after photobleaching (“FRAP”)

Inhibition of gap junction-mediated cell-cell communication might be a mechanism for several types of cellular dysfunctions, including tumor promotion. Although many different assays have been designed to measure gap junction-mediated intercellular communication, we applied a new technique, termed Fluorescence Redistribution After Photobleaching (FRAP), to assess the ability of a known tumor promoter, 2,2, 4,4, 5,5-hexabromobiphenyl (245-HBB), to inhibit cell-cell communication in a concentration-dependent manner. WB-F344 (rat epithelial) cells were plated at low density, exposed to noncytotoxic concentrations of 1, 5, or 20 µg 245-HBB/ml medium, and stained with 6-carboxyfluorescein diacetate. Single cells in pairs or clusters of touching cells in each exposure group were examined with FRAP. The results revealed an inverse correlation between the degree offluorescence redistribution in photobleached cells and the concentration of 245-HBB. Therefore, FRAP appears to be a sensitive and rapid technique for determining complete or partial inhibition of chemically induced intercellular communication in vitro. These results also provide further evidence for the ability of 245-HBB to inhibit gap junction-mediated cell-cell communication in a concentration-dependent manner.Abbreviations 6-CFDA 6-carboxyfluorescein diacetate - FRAP fluorescence redistribution after photobleaching - 245-HBB 2,2, 4,4, 5,5-hexabromobiphenyl Michigan Agricultural Experiment Station journal article No. 12531.     

Über das Aktionsspektrum der phototaktischen Reaktionen von Euglena

Zusammenfassung Bei Euglena gracilis zeigt das Aktionsspektrum der positiv-phototaktischen Reaktion, gemessen an Reizstärken, die eben über der Schwelle liegen, ein Hauptmaximum bei etwa 490–500 m, ein Nebenmaximum wurde bei etwa 415–430 m gefunden.Das Aktionsspektrum für die negativ-phototaktische Reaktion, gemessen an den Reizstärken, die eben über der Schwelle für diesen Reaktionstyp liegen, hat sein Hauptmaximum bei etwa 415 m; es stimmt ungefähr mit dem eben erwähnten Nebenmaximum der positivphototaktischen Reaktion überein.Die Empfindlichkeitsgrenze zum Langwelligen liegt für beide Reaktionstypen bei etwa 550 m.Die Befunde legen folgende Deutung nahe: Die negativ-phototaktische Reaktion erfolgt ohne Mitwirkung des Stigmas rein phobisch. Die positiv-phototaktische Reaktion ist an die gleiche strahlungsabsorbierende Substanz gebunden wie die negative, für sie ist dabei aber eine periodische Verdunkelung dieser Photoreceptorsubstanz durch die photochemisch nicht wirksamen Carotinoide des Stigmas erforderlich. Wo diese Verdunkelung wegen geringer Absorption der Stigma-Carotinoide (etwa bei 400 m) nicht möglich ist, kann nur die negative (phobotaktische) Reaktion entstehen. Wo umgekehrt zwar das Stigma absorbiert (etwa zwischen 550–600 m), nicht aber die lichtempfindliche Substanz des Photoreceptors, sind beide Reaktionen ausgeschlossen.     

Simultaneous overproduction of extracellular endoglucanase and intracellular β-glucosidase by genetically engineered Bacillus subtilis

Summary Simultaneous overproduction of intracellular -glucosidase and extracellular endoglucanase was attempted by constructing two artificial operon systems comprising the -glucosidase-endoglucanase gene(E) or the endoglucanase--glucosidase gene(E) under the control of a strong engineered promoter, BJ27U88 and expressing them in Bacillus subtilis DB104. Two artificial operon systems contained 30 bp or 5 bp gap between the termination codon of the upstream gene and the SD sequence of the downstream gene, respectively. These operon systems were expressed well in B. subtilis and overproduced the -glucosidase cell extract as well as the endoglucanase supernatant. The level of expression in the operon system was almost the same as that in a single expression system.     

Selective amidation of carboxyl groups of the intermolecular contact regions of hemoglobin S: Structural aspects

Carboxyl groups of HbS are readily activated by water-soluble carbodiimide atpH 6.0 and room temperature. These o-acylurea intermediates (activated carboxyl) are accessible for nucleophilic attack by amines. With glycine ethyl ester, the amidation is very selective for the -carboxyl of Glu-43() and more than 65% of the glycine ethyl ester incorporated is on this carboxyl group. In contrast, glucosamine derivatizes the -carboxyl group of Glu-22() as well as that of Glu-43() to nearly the same degree. However, the total amidation of HbS by glucosamine is lower than that with glycine ethyl ester. The differential selectivity of the two amines is apparently related to the differences in the microenvironment of the -carboxyl groups of Glu-22() and Glu-43(), which either facilitates or refracts the aminolysis of the activated carboxyl with the two amines to different degrees. The carboxyl groups of isolated -chain exhibit a higher reactivity for amidation with glycine ethyl ester than does the tetramer. The carboxyl groups of Glu-22() and Glu-43() and that of Asp-47() are all activated by carbodiimide suggesting that the higherpK_a of these carboxyl groups (facilitating the activation) is a property of tertiary interaction of the polypeptide chain. The interaction of the -chain with -chain, i.e., generation of the quaternary interactions, reduces overall reactivity of the carboxyl groups of the protein. The higher selectivity of hemoglobin S for amidation at Glu-43() with glycine ethyl ester compared with that of isolated -chain appears to be primarily a consequence of decreased amidation at sites other than at Glu-43().