The Structure and Composition of Bead-like Projections on Sitka Spruce Callus Cells Formed during Grafting and in Culture

Projections at the cell wall surface in callus formed at thegraft interface and in vitro in Picea sitchensis (Bong.) Carrhave been examined using light and electron microscopy. Thesestructures range in size from 1 to 5 µm in diameter, andconsist of a homogeneous matrix enclosing one or more fibrillar/vesicularcores. Histochemical and cytochemical studies have shown thatthe homogeneous matrix is made up of a mixture of pectins, carbohydrate,protein and fatty acids, while the fibrillar/vesicular componentis mainly carbohydrate and pectins. The possible functions ofthese structures are discussed on the bases of their composition.Copyright1993, 1999 Academic Press Picea sitchensis, callus, cell-wall projections, histochemistry, cytochemistry     

Leaf Nucleic Acids:: II. METABOLISM DURING SENESCENCE AND THE EFFECT OF KINETIN

The nucleic acids in the green and in the senescent leaves ofthree types of plant have been studied. High and low molecularweight RNA of the chloroplast is not present in senescent leavesof Xanthium pensylvanicum, but both cytoplasmic and chloroplasticfractions are found in yellow leaves of Vicia faba and Nicotianatabacum. RNA is more rapidly degraded than DNA in the leavesof these plants when they are detached, and kinetin treatmenttemporarily arrests the loss of chlorophyll and nucleic acid.Once X. pensylvanicum leaves are yellow and senescent they cannotbe re-greened, whereas those of Nicotiana spp., and to someextent those of V. faba, can be rejuvenated. We suggest thatthe retention of chloroplast RNA in yellow leaves may be a majorfactor determining their ability to re-green and that the patternof organelle senescence prior to the first stages of leaf autolysisand dehydration is species-specific.     

Observations on the Senescence of a Mutant Non-yellowing Genotype of Phaseolus vulgaris L.

A non-yellowing mutant of Phaseolus vulgaris L. was used toinvestigate factors involved in chlorophyll breakdown duringfoliar senescence. The mutant showed physiological changes similarto those of the normal yellowing type during senescence exceptthat leaf chlorophyll did not decline. Transmission electronmicroscope studies did not reveal appreciable differences inchloroplast ultrastructure between the two genotypes, suggestingthat chloroplast membrane integrity was not the factor preventingchlorophyll degradation in the mutant. However, the lack ofplastoglobuli in senescent mutant chloroplasts suggested thatthe lipid environment may be different from that of senescentnormal chloroplasts. Banding patterns of total soluble protein,resolved by sodium dodecyl sulphate-poly aery lamide gel electrophoresisshowed few, if any, differences between mature non-senescentnormal and mutant leaves; however, bands at 14 kD and 58 kDdiminished in senescent normal leaves, but remained in senescentmutant non-yellowing leaves. Key words: Non-yellowing mutant, Phaseolus vulgaris, senescence, chlorophyll degradation     

Production of reactive oxygen species in Arabidopsis thaliana cell suspension cultures in response to an elicitor from Fusarium oxysporum: implications for basal resistance

The present understanding of ROS generation in the defence responseof Arabidopsis thaliana is reviewed. Evidence suggests thatthe apoplastic oxidative burst generated during basal resistanceis peroxidase-dependent. The ROS generated during this basalresistance may serve to activate NADPH oxidase during the R-gene-mediatedhypersensitive response. The processes involved in the productionof reactive oxygen species in A. thaliana cell suspension culturesin response to an elicitor from Fusarium oxysporum are investigatedin the present work. This system appears analogous to the productionof ROS during the basal resistance response in French bean,which is peroxidase-dependent. A panel of modulators effectivein other pathogen elicitor and plant cell systems has been usedto investigate the Arabidopsis signalling pathways and the plantcell responses involved. Thus as in other systems, an earlycalcium influx into the cytosolic compartment, a rapid effluxof K⁺ and Cl^–, and extracellular alkalinization of elicitedcell cultures has been found. However the alkalinization isnot sufficient to stimulate the apoplastic oxidative burst byitself, unlike in French bean, although vectorial ion fluxesare needed. A secretory component which is sensitive to monensinand N-ethylmaleimide and insensitive to brefeldin A may alsobe necessary for the release and provision of substrates forperoxidase-dependent generation of H₂O₂. Key words: Arabidopsis thaliana, calcium, elicitation, hydrogen peroxide, oxidative burst, secretion     

Gastrulation in Calcareous Sponges: In Search of Haeckel's Gastraea

Haeckel's studies of development in calcareous sponges (1872)led him to develop the "Gastraea Theory," which proposes thatthe ancestral mode of germ layer formation, or gastrulation,was by invagination to produce a functional gut. His observationsthat gastrulation in the Calcarea occurs by invagination ofa ciliated larva upon settlement and metamorphosis were supportedby remarkable photomicrographs of the stage by Hammer in 1908.Although no later work found the same stage, these conceptsare repeated in texts today. We have re-examined embryogenesisand metamorphosis in Sycon sp. cf. S. raphanus in order to understandwhen gastrulation occurs. Almost all larvae settle on theirciliated anterior pole and metamorphose into a bilayered juvenilewhose interior cells rapidly differentiate into choanocytesand other cells of the young sponge. After a four-year searchwe have found the transitory stage shown by Hammer in whichthe anterior cells invaginate into the posterior half of thelarva. The hole closes and it is not until some days later thatthe sponge forms an osculum at its apical pole. To understandwhether invagination comprises gastrulation and if the holecan be considered to be a blastopore we have carried out a reviewof the literature dealing with this brief moment in calcaroneansponge development. Despite the intrigue of this type of metamorphosis,we conclude that gastrulation occurs earlier, during formationof the two cellular regions of the larva, and that metamorphosisinvolves the reorganization of these already differentiatedregions. Considering the pivotal position occupied by the Calcareaas the possible sister-group to all other Metazoa, these resultscall for a reassessment of germ layer formation and of the relationshipsof the primary germ layers among basal metazoan phyla.     

THE BIOLOGY AND FUNCTIONAL MORPHOLOGY OF MYTILOPSIS SALLEI (RECLUZ) (BIVALVIA: DREISSENACEA) FOULING VISAKHAPATNAM HARBOUR, ANDHRA PRADESH, INDIA

Mytilopsis sallei (Récluz) (Dreissenacea) has been introducedinto Visakhapatnam harbour, India. The normal distribution ofthe species is Central America and it is here contended thatthe species has been introduced into the Indo-Pacific via thePanama canal (opened in 1915) attached to the hulls of ships.M. allyneana (Hertlein & Hanna) with a distribution restrictedto Fiji is considered synonymous with M. sallei. The literature on M. sallei is reviewed and it is shown thatthe species has wide temperature, salinity and oxygen tolerancesand thus transport across a variety of water masses is entirelyfeasible. The species also possesses a fast rate of growth,a high fecundity and matures early. It is an opportunistic specieswhich free from the restraints that normally hold it in checkin its natural environment, poses a serious fouling threat toharbours throughout the Indo-Pacific and to the endemic fauna. The morphological adaptations of M. sallei are also describedand compared with those of Dreissena polymorpha (Pallas) inparticular and other fouling bivalves in general. The speciesis ideally adapted to life in turbid estuaries experiencingwide variations in temperature, salinity, oxygen and turbulence.A warning against further introductions is given. (Received 14 December 1979;     

Studies on the growth and nitrogen assimilation of the bloom dinoflagellate Gyrodinium aureolum Hulburt

Growth experiments were carried out with laboratory culturesof Gyrodinium aureolum. Physiological studies on CO₂ fixationand nitrogen nutrition were undertaken with a natural populationdominated by the same organism. These studies suggested thatthe population was in a late stage of bloom development andsuffering from a degree of nitrogen limitation. The relevanceof the ammonium enhancement of dark CO₂ fixation as a techniquefor determining the nitrogen status of phytoplankton is discussedas are the difficulties involved in extrapolating results fromlaboratory studies to field situations.     

The Effect of Adenine and Mevalonic acid on the Endogenous Cytokinins of a Cytokinin-dependent Strain of Soya Bean Callus

The combined application of 10^–6 M adenine and 10^–6M mevalonic acid to soya bean callus accelerated its growth.Two biologically active compounds that co-chromatographed withzeatin and isopentenyl adenine were extracted from this callus.Studies with labelled adenine and mevalonic acid indicated thatthe cytokinin-dependent soya bean callus incorporated only avery small amount of the radioactive precursors into the biologically-activecompounds, making it extremely difficult to determine whetherthese compounds were synthesized de novo or whether they aroseas by-products of tRNA turnover. As cytokinins do not accumulatein rapidly-growing cytokinin-dependent soya bean callus culturedon kinetin as a source of cytokinin it seems as if biosynthesisde novo occurs when the callus is supplied with adenine andmevalonic acid. Glycine max (L.) Merrill, soya bean, callus culture, adenine, mevalonic acid, endogenous cytokinins     

Synthesis and Characterization of Novel 1,3-Diaryltriazene-Substituted Sulfaguanidine Derivatives as Selective Carbonic Anhydrase Inhibitors: Biological Evaluation,in Silico ADME/T and Molecular Docking Study

Sulfonamide compounds known as human carbonic anhydrase (hCA) inhibitors are used in the treatment of many diseases such as epilepsy, antibacterial, glaucoma, various diseases. 1,3-diaryl-substituted triazenes and sulfaguanidine are used for therapeutic purposes in many drug structures. Based on these two groups, the synthesis of new compounds is important. In the present study, the novel 1,3-diaryltriazene-substituted sulfaguanidine derivatives ( SG1-13 ) were synthesized and fully characterized by spectroscopic and analytic methods. Inhibitory effect of these compounds on the hCA I and hCA II was screened as in vitro. All the series of synthesized compounds have been identified as potential hCA isoenzymes inhibitory with K_I values in the range of 6.44±0.74-86.85±7.01 nM for hCA I and with K_I values in the range of 8.16±0.40-77.29±9.56 nM for hCA II. Moreover, the new series of compounds showed a more effective inhibition effect than the acetazolamide used as a reference. The possible binding positions of the compounds with a binding affinity to the hCA I and hCA II was demonstrated by in silico studies. In conclusion, compounds with varying degrees of affinity for hCA isoenzymes have been designed and as selective hCA inhibitors. These compounds may be potential alternative agents that can be used to treat or prevent diseases associated with glaucoma and hCA inhibition.     

Purification and partial characterization of two biologically active compounds from the inflorescence of Sauromatum guttatum Schott (Araceae)

The active principle triggering the metabolic outburst of theappendices of certain arum lily species has been purified andpartially characterized. Although in the past it has been thoughtof as a single factor ("calorigen"), both paper chromatographyand DEAE-Sephadex ion exchange chromatography reveal that thereare two compounds (calorigen I and II) with calorigen activity.These can act alone or together, but show no synergism. Theyare low-molecular compounds which appear to be common constituentsof the inflorescence of Sauromatum guttatum Schott. CalorigenII is also present in the spadix of Arum maculatum L., Arumdioscoridis Sibth. & Sm., and Dracunculus vulgaris Schott.Calorigen I absorbs UV at 242, 267 and 278 nm, calorigen IIat 229 and 295 nm. No spectral changes are noted in alkalineenvironment, whereas at low pH-values there is a shift of theabsorption peaks of both compounds towards longer wavelengths.Calorigen II displays a strong blue fluorescence and has a maximalemission at 410 nm when activated at 295 nm. The possible mechanismof calorigen action is discussed. ¹ Present address: Department of Biochemistry, University ofWashington, Seattle, Washington 98195, U.S.A. (Received August 26, 1974; )     

Involvement of Reactive Nitrogen and Oxygen Species (RNS and ROS) in Sunflower-Mildew Interaction

Nitric oxide (·NO) has been shown to participate in plantresponse against pathogen infection; however, less is knownof the participation of other NO-derived molecules designatedas reactive nitrogen species (RNS). Using two sunflower (Helianthusannuus L.) cultivars with different sensitivity to infectionby the pathogen Plasmopara halstedii, we studied key componentsinvolved in RNS and ROS metabolism. We analyzed the superoxideradical production, hydrogen peroxide content, L-arginine-dependentnitric oxide synthase (NOS) and S-nitrosoglutathione reductase(GSNOR) activities. Furthermore, we examined the location andcontents of ·NO, S-nitrosothiols (RSNOs), S-nitrosoglutathione(GSNO) and protein 3-nitrotyrosine (NO₂-Tyr) by confocal laserscanning microscopy (CLSM) and biochemical analyses. In thesusceptible cultivar, the pathogen induces an increase in proteinsthat undergo tyrosine nitration accompanied by an augmentationin RSNOs. This rise of RSNOs seems to be independent of theenzymatic generation of ·NO because the L-arginine-dependentNOS activity is reduced after infection. These results suggestthat pathogens induce nitrosative stress in susceptible cultivars.In contrast, in the resistant cultivar, no increase of RSNOsor tyrosine nitration of proteins was observed, implying anabsence of nitrosative stress. Therefore, it is proposed thatthe increase of tyrosine nitration of proteins can be considereda general biological marker of nitrosative stress in plantsunder biotic conditions.     

A Lectin from Leaves of Neoregelia flandria Recognizes D-Glucose, D-Mannose and N-Acetyl D-glucosamine, Differing from the Mannose-Specific Lectins of Other Monocotyledonous Plants

A mannose-specific lectin was isolated from leaves of Neoregeliaflandria, an ornamental plant that belongs to Bromeliaceae,a family of monocotyledons. The amino acid composition and molecularmass of the lectin were similar to those of mannose-specificlectins from other monocotyledons. However, in a test to examinethe inhibition of hemagglutination, it became apparent thatthe isolated lectin recognized D-glucose and N-acetyl D-glucosaminein addition to D-mannose, unlike mannose-specific lectins fromthe monocotyledons that have been reported to date. (Received May 17, 1996; Accepted August 19, 1996)     

Structure--function studies of oligosaccharides of recombinant human thyrotrophin by sequential deglycosylation and resialylation

Recombinant human thyrotrophin (rhTSH) contains oligosaccharidesterminating in -galactose-sialic acid, and had lower metabolicclearance and higher in vivo bioactivity compared to pituitaryhTSH, which has oligosaccharides terminating predominantly in-N-acetylgalactosamine-sulphate. Previous studies using completeremoval of the oligosaccharide chains showed an important rolefor the carbohydrate in the biological activity of the hormone.In the present study, we have determined the contribution ofthe individual monosaccharides to hormonal activity by sequentialdeglycosylation of rhTSH using exoglycosidases. We have alsoinvestigated the effect of resialylation of desialylated rhTSHusing sialyltransferases. Sequential removal of sialic acid,galactose or N-acetylglucosamine resulted in a > 10-foldincrease in the in vitro bioactivity of rhTSH. The metabolicclearance of the derivatives was faster than that of intacthormone, but agalacto-rhTSH was cleared slower than asialo-rhTSH.However, the in vivo bioactivity decreased progressively witheach monosaccharide removal. The increased cyclic AMP-stimulatingactivity, increased metabolic clearance and the decreased invivo biologic activity were all reversed by resialylation ofthe terminal galactose residues. These results indicate thatthe in vitro, as well as the in vivo, bioactivities of rhTSHare modulated by terminal sialylation. The effects of sequentialdeglycosylation on the in vitro activity of rhTSH are differentfrom those reported earlier for human chorionic gonadotrophin.Thus, modification of the oligosaccharides by glycosidases andglycosyltransferases can be used as a powerful tool to delineatethe function of carbohydrate in glycoproteins and to engineermore potent hormone analogues with a longer half-life and/orhigher bioactivity. deglycosylation exoglycosidases oligosaccharides recombinant thyrotrophin sialyltransferases     

THE REPRODUCTIVE CYCLE AND SEXUALITY OF THE GREEN MUSSEL PERNA VIRIDIS (L.) (BIVALVIA: MYTILACEA) IN VICTORIA HARBOUR, HONG KONG

The reproductive cycle and sexuality of the green mussel Pernaviridis (L.) in Victoria Harbour, Hong Kong, were investigatedfrom July 1982 to May 1984. Histological studies showed thatthe cyde could be divided into 4 stages occurring in a highlyseasonal pattern. The very low (<0.1%) occurrence of functionalhermaphrodites indicates that P. viridis is gonochoristic. Pastreports of protandric rhythmic hermaphroditism may have resultedfrom reliance on fluctuations in sex ratio and sex identificationby gonad colour. These criteria are, however, considered invalidfor P. viridis because differences exist between the developmentalpatterns of the male and female gonads and, further, colourof the female gonad at the early proliferation stage resemblesthe male. Temperature was found to correlate positively withgonad development and with the lower threshold at 24°C.Temperature was not, however, limiting at some sites in HongKong. A joint Principal Component Analysis-Stepwise multipleregression procedure suggested that hypertrophication mightinhibit gonadal development in Victoria Harbour (Received 2 November 1987; accepted 30 December 1987)     

Cytogenetic Studies in Cochlearia L. The chromosomal homogeneity within both the 2n = 12 diploids and the 2n = 14 diploids and the cytogenetic relationship between the two chromosome levels

A cytogenetic study has been carried out within the diploidCochleariae. This has demonstrated that all the 2n = 12 taxaare chromosomally homogeneous, as are those with 2n = 14. Itis therefore suggested that the 2n = 12 group be recognisedas C pyrenaica DC, with the morphologically distinct specimensfrom France and Spain at present regarded as C. aestuaria (Lloyd)Heywood being given subspecific rank in C. pyrenaica. It isalso suggested that all the 2n = 14 taxa be regarded as C groenlandicaL. Some doubt is expressed on the existence of C. scotica Druceas a species, as all the specimens of this taxon examined provedto be tetraploid with 2n = 24. Genomic analysis has shown thatthe 2n = 14 group is a primary tetrasomic derivative of the2n = 12 group.     

Comparing peripheral olfactory coding with host preference in the rhagoletis species complex

Recent studies have shown that flies from sympatric populationsof Rhagoletis pomonella infesting hawthorn, apple, and floweringdogwood fruit can distinguish among unique volatile blends identifiedfrom each host. Analysis of peripheral chemoreception in Rhagoletisflies suggests that changes in receptor specificity and/or receptorneuron sensitivity could impact olfactory preference among thehost populations and their hybrids. In an attempt to validatethese claims, we have combined flight tunnel analyses and singlesensillum electrophysiology in F₂ and backcross hybrids displayinga variety of behavioral phenotypes. Results show that differencesin peripheral chemoreception among second-generation adultsdo not provide a direct correlation between peripheral codingand olfactory behavior. We conclude that either the plasticityof the central nervous system in Rhagoletis can compensate forsignificant alterations in peripheral coding or that peripheralchanges present subtle effects on behavior not easily detectablewith current techniques. The results of this study imply thatthe basis for olfactory behavior in Rhagoletis has a complicatedgenetic and neuronal basis, even for populations with a recentdivergence in preference.     

A Step in the Biosynthesis of Gibberellins that Is Controlled by the Mutation in the Semi-Dwarf Rice Cultivar Tan-Ginbozu

Genetic analysis and a comparison of endogenous levels of gibberellinsbetween the semi-dwarf rice cultivar Tan-ginbozu and the correspondingnormal cultivar Ginbozu have confirmed that Tan-ginbozu is agibberellin deficient mutant and that the semi-dwarfism of Tan-ginbozuis controlled by a single recessive gene. A step in the biosynthesisof gibberellins that is blocked by the mutation in Tan-ginbozuhad been considered to be the synthesis of ent-kaurene or anearlier step. However, the rate of production of ent-kaureneby Tan-ginbozu was almost the same as that by Ginbozu. By contrast,accumulation of only a small amount of ent-kaurene was detectedin Tan-ginbozu, and the amount that accumulated was similarto that in Ginbozu that had been treated with 6.9 x 10^-8 M uniconazole-P(an effective inhibitor of three oxidative steps in the pathwayfrom ent-kaurene to ent-kaurenoic acid via entkaurenol and ent-kaurenal).The height of the treated Ginbozu plants was reduced to thesame as that of Tanginbozu plants. Resembling Tan-ginbozu plants,Ginbozu plants that had been treated with uniconazole-P respondedwell to ent-kaurenoic acid and slightly to ent-kaurene and ent-kaurenol.Since the growth-promoting activity of enf-kaurenal in Tan-ginbozuwas similar to that of ent-kaurene, our results suggest thatthe mutation in Tan-ginbozu blocks the three oxidative stepswhereby ent-kaurene is converted to ent-kaurenoic acid. (Received June 9, 1995; Accepted February 15, 1996)     

Estimation of the mean function with panel count data using monotone polynomial splines

We study nonparametric likelihood-based estimators of the meanfunction of counting processes with panel count data using monotonepolynomial splines. The generalized Rosen algorithm, proposedby Zhang & Jamshidian (2004), is used to compute the estimators.We show that the proposed spline likelihood-based estimatorsare consistent and that their rate of convergence can be fasterthan n^1/3. Simulation studies with moderate samples show thatthe estimators have smaller variances and mean squared errorsthan their alternatives proposed by Wellner & Zhang (2000).A real example from a bladder tumour clinical trial is usedto illustrate this method.     

Carboxyarabinitol 1-Phosphate Phosphatase from Leaves of Phaseolus vulgaris and Other Species

Carboxyarabinitol 1-phosphate (CA1P) phosphatase activity occurredin leaves of 10 species examined, with the highest activityin leaves of Phaseolus vulgaris. Enzyme was purified from P.vulgaris 1,580-fold to a final specific activity of 6.1 µmolmin^–1 (mg protein)^–1. Structural characteristicsof positive effectors and substrate analogs for the CA1P phosphatasereaction were examined. Positive effectors were compounds thatcontained one phosphate group in close proximity to a secondphosphate or a carboxyl group (e.g. 2-phosphoglycolate, pyrophosphate,3-phosphoglycerate, and carboxyethylphosphonic acid). Many ofthe activators are structurally quite similar to CA1P, but werenot used as substrates. In addition to the natural substrateCA1P, carboxypentitol and carboxyhexitol bisphosphates wereshown to be good substrates (e.g. carboxyarabinitol bisphosphateand carboxymannitol bis-phosphate). A substrate arabinitol configuration(R) was preferred at C-2, and reactivity was lost when a hydroxymethylgroup was substituted for the carboxyl group. Despite structuralsimilarities to positive effectors, none of the tested reactionsubstrates could activate the enzyme. (Received November 11, 1996; Accepted February 3, 1997)     

Early Pleistocene glacial-interglacial radiolarian assemblages from the eastern equatorial Pacific

Polycystine radiolaria from ODP Hole 677A in the eastern equatorialPacific were examined at isotopically identified Early Pleistoceneglacial maxima and minima. Two distinct radiolarian assemblagesare recognized, characterizing glacial and interglacial optima.The Glacial Assemblage is characterized by high abundances ofTheocalyptra davisiana, Botryostrobus auritus, Anthocyrtidiumzanguebaricuim andHexacontium enthacanthum. The InterglacialAssemblage is characterized by Tetrapyle octacantha, Octapylestenozoa and Theocorythium vetulum. A comparison of these fossilassemblages with modern radiolarian distribution suggests thatthe Glacial Assemblage represents intensified upwelling of coldadvected water via the Eastern Pacific Boundary Current, whilstthe Interglacial Assemblage indicates climatic ameliorationin the eastern equatorial Pacific, with the prevalence of warm(>21C) tropical/subtropical surface waters. The recognitionof these radiolarian assemblages could be successfully appliedto studies of adjacent east Pacific areas where other palaeoecologicalindicators are lacking.