Publications

biosynthesis of 12α-and 13-hydroxylated gibberellins in a cell-free system from cucurbita maxima endosperm and the identification of new endogenous gibberellins.	gibberellin (ga) biosynthesis in cell-free systems from cucurbita maxima l. endosperm was reinvestigated using incubation conditions different from those employed in previous work. the metabolism of ga12 yielded ga13, ga43 and 12α-hydroxyga43 as major products, ga4, ga37, ga39, ga46 and four unidentified compounds as minor products. the intermediates ga15, ga24 and ga25 accumulated at low protein concentrations. the structure of the previously uncharacterised 12α-hydroxyga43 was inferred from it ...	1993	24178490
purification, characterization, and complete amino acid sequence of a trypsin inhibitor from amaranth (amaranthus hypochondriacus) seeds.	a protein proteinase inhibitor was purified from a seed extract of amaranth (amaranthus hypochondriacus) by precipitation with (nh4)2so4, gel-filtration chromatography, ion-exchange chromatography, and reverse-phase high-performance liquid chromatography. it is a 69-amino acid protein with a high content of valine, arginine, and glutamic acid, but lacking in methionine. the inhibitor has a relative molecular weight of 7400 and an isoelectric point of 7.5. it is a serine proteinase inhibitor that ...	1993	8290633
identification of sulphur-rich proteins which resist rumen degradation and are hydrolysed rapidly by intestinal proteases.	several proteins with high proportions of s-containing essential amino acids were incubated in sheep rumen fluid in vitro and their rate of digestion was examined by sodium dodecyl sulphate-polyacrylamide-gel electrophoresis. the s-rich proteins rice prolamin (10 kda), maize zein (10 kda) and the 3.2 kda pumpkin (cucurbita maxima l.) trypsin inhibitor-1 (cmti-1) were highly resistant to rumen fluid degradation, relative to control proteins of known degradation rate (casein, bovine serum albumin ...	1994	7827006
toxicologic evaluation of acute and subacute oral administration of cucurbita maxima seed extracts to rats and swine.	the extract prepared from dried seeds of cucurbita maxima was administered to rats and pigs. following a single dose or 4 weeks of daily oral administration, the extract produced no changes in serum glucose, urea, creatinine, total protein, uric acid, got, gpt, ldh or blood counts. urine analysis (urea, uric acid, creatinine, total protein, na and k), as well as histopathological investigation, showed no abnormalities. these results taken as a whole indicate that the seeds of c. maxima as used i ...	1994	7967649
new active analogues of cucurbita maxima trypsin inhibitor iii (cmti-iii) modified in the non-contact region.	four new analogues of trypsin inhibitor cmti-iii(3-28) = [desarg1,desval2,desgly29]cmti-iii which was recently shown to be fully active, were synthesized by the solid-phase method. the introduction of glycine in position 9 (peptide 1) and gly-pro-gly (peptide 2) and gly-pro-asn (peptide 3) in the regions 17-19 and 23-25, respectively, did not change the antitrypsin activity of all modified peptides. all of these substitutions are presumed to be outside the trypsin-binding loop as judged from the ...	1994	8003251
analogues of cucurbita maxima trypsin inhibitor iii (cmti-iii) with elastase inhibitory activity.	three new cmti-iii analogues containing the val residue in the reactive site (position 5) were synthesized by the solid-phase method. the analogues displayed an elastase inhibitory activity. it is shown that the removal of the n-terminal arg residue and the introduction of the gly-pro-gln tripeptide in the region 23-25 decreases the antielastase activity by two orders of magnitude. the removal of the disulfide bridge in positions 16-28 and the substitution of ala for cys16 and gly for cys28 decr ...	1994	8060538
expression cloning of a gibberellin 20-oxidase, a multifunctional enzyme involved in gibberellin biosynthesis.	in the biosynthetic pathway to the gibberellins (gas), carbon-20 is removed by oxidation to give the c19-gas, which include the biologically active plant hormones. we report the isolation of a cdna clone encoding a ga 20-oxidase [gibberellin, 2-oxoglutarate:oxygen oxidoreductase (20-hydroxylating, oxidizing) ec 1.14.11.-] by screening a cdna library from developing cotyledons of pumpkin (cucurbita maxima l.) for expression of this enzyme. when mrna from either the cotyledons or the endosperm was ...	1994	8078921
single peptide bond hydrolysis/resynthesis in squash inhibitors of serine proteinases. 1. kinetics and thermodynamics of the interaction between squash inhibitors and bovine beta-trypsin.	the substrate and inhibitory parameters are described for the interaction between cucurbita maxima trypsin inhibitor i (cmti i) and bovine beta-trypsin. the data are fully consistent with the reactive site hypothesis and the standard mechanism proposed for the protein inhibitor-serine proteinase interaction. the second-order association rate constant (k(on)) for the interaction of the intact inhibitor and trypsin is high, above 10(6) m-1 s-1. the same value is only 22-fold lower for the reactive ...	1994	8286341
aluminum-induced rapid root inhibition and changes in cell-wall components of squash seedlings.	growth of squash (cucurbita maxima duch.) roots was significantly inhibited by 1 mm alcl3 as early as 1 h after the treatment. the growth inhibition was confined to the elongating zone (1-6 mm from the root tip). chemical analysis of cell-wall polysaccharides from roots revealed that aluminum increased pectin, hemi-cellulose, and cellulose contents after 3 h of treatment. the effect of aluminum on pectin content was found in the elongating zone including the root tip, whereas change in cellulose ...	1994	12232377
organization and characterization of cucurbita phloem lectin genes.	the phloem of pumpkin and squash contains a dimeric chitin-binding lectin called pp2 (phloem protein 2). we have isolated three genomic clones from pumpkin (cucurbita maxima duch.) that encoded pp2. one clone, lambda gpc13-1, contained two pp2 genes that were 99.8% identical over a region of 3055 nucleotides. this conserved region included 1922 bp of 5' non-coding sequence, 844 bp of protein coding sequence (including two introns), and 289 bp of 3' non-coding sequence. to examine the conservatio ...	1994	8000002
single peptide bond hydrolysis/resynthesis in squash inhibitors of serine proteinases. 2. limited proteolysis of curcurbita maxima trypsin inhibitor i by pepsin.	porcine pepsin hydrolyzes the leu7-met8 (p2'-p3') peptide bond in cucurbita maxima trypsin inhibitor i (cmti i) in the ph range 2.0-4.8. the reaction proceeds to equilibrium between intact cmti i and its cleaved form. the ph-independent value of the equilibrium constant (khyd0 = 0.78) indicates that both forms of the inhibitor have similar gibbs energies. the ph dependence of this constant shows that the peptide bond hydrolysis does not perturb ionization constants of any preexistent groups. the ...	1994	8286342
characterization and sequencing of cdna clone encoding the phloem protein pp2 of cucurbita pepo.	direct n-terminal amino acid sequencing of the phloem protein 2 (pp2) from 3-month old cucurbita pepo l. (pumpkin), purified by sds-page and blotted onto pvdf membrane, showed that the protein had a blocked n-terminus. however, after in situ cleavage of the polypeptide in a gel slice by cyanogen bromide, 75 residues of sequences on two cyanogen bromide fragments were determined. an oligonucleotide probe based on this amino acid sequence was used to screen a cdna library, constructed from mrna of ...	1994	8111014
purification and partial amino-acid sequence of gibberellin 20-oxidase from cucurbita maxima l. endosperm.	gibberellin (ga) 20-oxidase was purified to apparent homogeneity from cucurbita maxima endosperm by fractionated ammonium-sulphate precipitation, gel-filtration chromatography and anion-exchange and hydrophobic-interaction high-performance liquid chromatography (hplc). average purification after the last step was 55-fold with 3.9% of the activity recovered. the purest single fraction was enriched 101-fold with 0.2% overall recovery. apparent relative molecular mass of the enzyme was 45 kda, as d ...	1994	7765793
ecology of nematodes under influence of cucurbita spp. and different fertilizer types.	in a field study conducted in georgia, cucurbit plants with high (cucurbita andreana) and low (cucurbita maxima) concentrations of cucurbitacins were used in combination with two types of fertilizers to investigate their effects on the community of soil nematodes. ecological measures of soil nematode community structure such as total nematode abundance, number of genera, trophic diversity, trophic group proportions, fungivore/bacterivore ratio, and modified maturity index were assessed and compa ...	1995	19277330
purification and properties of ent-kaurene synthase b from immature seeds of pumpkin.	ent-kaurene synthase b (ksb) was purified 291-fold from a crude enzyme preparation from endosperm of pumpkin (cucurbita maxima l.). separation of ent-kaurene synthase a and ksb was achieved by hydrophobic interaction chromatography. the fractions containing ksb activity were further purified by diethylaminoethyl, phenyl, and hydroxyapatite column chromatography. using sodium dodecyl phosphate-polyacrylamide gel electrophoresis, the purest enzyme preparation showed a major band at an apparent mol ...	1995	12228665
purification of a trypsin inhibitor (pfti) from pumpkin fruit phloem exudate and isolation of putative trypsin and chymotrypsin inhibitor cdna clones.	the major trypsin inhibitor from pumpkin (cucurbita maxima cv supermarket hybrid) fruit phloem exudate was purified by affinity and reverse phase chromatography. the protein has a molecular weight of approximately 8100 by sds-page and is blocked at the n-terminal serine. following sequencing of a cnbr fragment, 3'- and 5'-race were used to isolate full length cdnas corresponding to a trypsin inhibitor and to two chymotrypsin inhibitors. the three genes are similar, both in their translated and n ...	1995	7662170
three-dimensional solution structure of cucurbita maxima trypsin inhibitor-v determined by nmr spectroscopy.	the solution structure of cucurbita maxima trypsin inhibitor-v (cmti-v), which is also a specific inhibitor of the blood coagulation protein, factor xiia, was determined by 1h nmr spectroscopy in combination with a distance-geometry and simulated annealing algorithm. sequence-specific resonance assignments were made for all the main-chain and most of the side-chain hydrogens. stereospecific assignments were also made for some of the beta-, gamma-, delta-, and epsilon-hydrogens and valine methyl ...	1995	7711040
growth of cucurbita maxima l. plants in the presence of the cycloartenol synthase inhibitor u18666a.	squash, like other cucurbitaceae, have unique sterol profiles that offer an excellent opportunity to examine the relationship between sterol biosynthesis and plant growth. to determine the effect of sterol biosynthesis inhibition on squash growth, cucurbita maxima seedlings with and without cotyledons were subjected to increasing concentrations of the cycloarternol synthase (ec 5.4.99.8) inhibitor 3 beta-(2-diethylaminoethoxy)androstenone (u18666a). inhibition of shoot growth was concentration-d ...	1995	7791534
synthesis, cloning and expression in escherichia coli of a gene coding for the met8-->leu cmti i--a representative of the squash inhibitors of serine proteinases.	a chemically synthesized gene coding for a cucurbita maxima trypsin inhibitor modified at position p'3 (met8-->leu cmti i), i.e. at the third position downstream of the reactive site bond (arg5-ile), was cloned into a derivative of the plasmid paed4 that utilizes a t7 expression system. the gene was expressed in escherichia coli as a fusion protein that accumulates in inclusion bodies. after reduction and cnbr cleavage of the fusion protein followed by oxidative refolding and reverse-phase hplc, ...	1995	8543044
solution conformations of proline rings in proteins studied by nmr spectroscopy.	three different conformations of proline rings in a protein in solution, up, down and twist, have been distinguished, and stereospecific assignments of the pyrrolidine beta-, gamma- and delta-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 alpha-beta 3, beta 3-gamma 3, beta 2-gamma 2, gamma 2-delta 2, and gamma 3-delta 3 (2.3 a) are shorter than those in the pairs alpha-beta ...	1995	8589600
changing the inhibitory specificity and function of cucurbita maxima trypsin inhibitor-v by site-directed mutagenesis.	cucurbita maxima trypsin inhibitor-v (cmti-v) is also a specific inhibitor of human blood coagulation factor beta-factor xiia. a recombinant version of cmti-v has allowed probing of roles of individual amino acid residues including the reactive site residue, lysine (p1), by site-directed mutagenesis. the k44r showed at least a 5-fold increase in inhibitory activity toward human beta-factor xiia, while there was no change toward bovine trypsin. this result demonstrates that beta-factor-xiia prefe ...	1995	7864895
reactive-site hydrolyzed cucurbita maxima trypsin inhibitor-v: function, thermodynamic stability, and nmr solution structure.	reactive-site (lys44-asp45 peptide bond) hydrolyzed cucurbita maxima trypsin inhibitor-v (cmti-v) was prepared and characterized: in comparison to the intact form, cmti-v exhibited markedly reduced inhibitory properties and binding affinities toward trypsin and human blood coagulation factor xiia. the equilibrium constant of trypsin-catalyzed hydrolysis, khyd, defined as [cmti-v*]/[cmti-v], was measured to be approximately 9.4 at 25 degrees c (delta g degrees = -1.3 kcal.mol-1). from the tempe ...	1995	7547948
the ga5 locus of arabidopsis thaliana encodes a multifunctional gibberellin 20-oxidase: molecular cloning and functional expression.	the biosynthesis of gibberellins (gas) after ga12-aldehyde involves a series of oxidative steps that lead to the formation of bioactive gas. previously, a cdna clone encoding a ga 20-oxidase [gibberellin, 2-oxoglutarate:oxygen oxidoreductase (20-hydroxylating, oxidizing), ec 1.14.11.-] was isolated by immunoscreening a cdna library from liquid endosperm of pumpkin (cucurbita maxima l.) with antibodies against partially purified ga 20-oxidase. here, we report isolation of a genomic clone for ga 2 ...	1995	7604047
isolation and expression of three gibberellin 20-oxidase cdna clones from arabidopsis.	using degenerate oligonucleotide primers based on a pumpkin (cucurbita maxima) gibberellin (ga) 20-oxidase sequence, six different fragments of dioxygenase genes were amplified by polymerase chain reaction from arabidopsis thaliana genomic dna. one of these was used to isolate two different full-length cdna clones, at2301 and at2353, from shoots of the ga-deficient arabidopsis mutant ga1-2. a third, related clone, yap169, was identified in the database of expressed sequence tags. the cdna clones ...	1995	7630935
antigenotoxic spinasterol from cucurbita maxima flowers.	the antigenotoxic constituent of squash flowers was isolated by solvent partitioning and repeated vacuum liquid chromatography. the micronucleus test, an in vivo method, was used to monitor the antigenotoxicity of the various fractions during the isolation process. isolate sqfwb2d from the chloroform extract of squash flowers is the most antigenotoxic isolate. it decreased the mutagenicity of tetracycline by 64.7% at a dosage of 100 mg/kg mouse. statistical analysis using kruskall wallis one-way ...	1996	8649469
molecular cloning and photoperiod-regulated expression of gibberellin 20-oxidase from the long-day plant spinach.	spinach (spinacia oleracea l.) is a long-day (ld) rosette plant in which stem growth under ld conditions is mediated by gibberellins (gas). major control points in spinach are the later steps of sequential oxidation and elimination of c-20 of c20-gas. degenerate oligonucleotide primers were used to obtain a polymerase chain reaction product from spinach genomic dna that has a high homology with ga 20-oxidase cdnas from cucurbita maxima l. and arabidopsis thaliana heynh. this polymerase chain rea ...	1996	8742334
molecular cloning and characterization of a cdna encoding the gibberellin biosynthetic enzyme ent-kaurene synthase b from pumpkin (cucurbita maxima l.).	the first committed step in the formation of diterpenoids leading to gibberellin (ga) biosynthesis is the conversion of geranylgeranyl diphosphate (ggdp) to ent-kaurene. ent-kaurene synthase a (ksa) catalyzes the conversion of ggdp to copalyl diphosphate (cdp), which is subsequently converted to ent-kaurene by ent-kaurene synthase b (ksb). a full-length ksb cdna was isolated from developing cotyledons in immature seeds of pumpkin (cucurbita maxima l.). degenerate oligonucleotide primers were des ...	1996	8771778
internal mobility of reactive-site-hydrolyzed recombinant cucurbita maxima trypsin inhibitor-v characterized by nmr spectroscopy: evidence for differential stabilization of newly formed c- and n-termini.	the solution structure and internal dynamics of the reactive-site (lys44-asp45 peptide bond) hydrolyzed form of recombinant cucurbita maxima trypsin inhibitor-v (rcmti-v) were characterized by the application of two-dimensional 1h-15n nmr methods to the uniformly 15n-labeled protein. the 1h-15n chemical shift correlation spectra of rcmti-v were assigned, and the chemical shift data were compared with those available for rcmti-v [liu, j., prakash, o., cai, m., gong, y., huang, y., wen, l., wen, ...	1996	8823186
synthesis of a squash-type protease inhibitor by gene engineering and effects of replacements of conserved hydrophobic amino acid residues on its inhibitory activity.	cucurbita maxima trypsin inhibitor i (cmti-i), a member of the squash-type protease inhibitor family, is composed of 29 amino acids and shows strong inhibition of trypsin by its compact structure. to study the structure-function relationship of this inhibitor using protein engineering methods, we constructed an expression system for cmti-i as a fused protein with porcine adenylate kinase (adk). a met residue was introduced into the junction of adk and cmti-i to cleave the fusion protein with cnb ...	1996	9010939
differential modulation of binding loop flexibility and stability by arg50 and arg52 in cucurbita maxima trypsin inhibitor-v deduced by trypsin-catalyzed hydrolysis and nmr spectroscopy.	the side chains of arg50 and arg52 iin cucurbita maxima trypsin inhibitor-v (cmti-v) anchor the binding loop to the scaffold region [cai, m., gong, y., kao, j.l-f., & krishnamoorthi, r. (1995) biochemistry 34, 5201-5211]. the consequences of these hydrogen-bonding and electrostatic interactions on the conformational flexibility and stability of the binding loop were evaluated by trypsin-catalyzed hydrolysis of cmti-v mutants, in which each of the arginines was individually replaced with ala, lys ...	1996	8664268
[color loss evaluation of a dietary formulation in function of temperature, storage time and packaging].	color loss of a dietetic formulae based on pumpkin (cucurbita maxima), rice, hen and vegetable oils was evaluated. the product was under an accelerated test at temperatures of 40, 50 and 70 degrees c for a month. the kinetic reaction of color loss (quality loss indicator) was obtained with n = 1 and ea = 11.5 kcal/mol. the product was also stored for sixth months at temperatures of 25, 30, 35 and 40 degrees c using glass containers, laminated and composite cans (non metallic). the color was meas ...	1996	9161461
solution structure and backbone dynamics of recombinant cucurbita maxima trypsin inhibitor-v determined by nmr spectroscopy.	the solution structure of recombinant cucurbita maxima trypsin inhibitor-v (rcmti-v), whose n-terminal is unacetylated and carries an extra glycine residue, was determined by means of two-dimensional (2d) homo and 3d hetero nmr experiments in combination with a distance geometry and simulated annealing algorithm. a total of 927 interproton distances and 123 torsion angle constraints were utilized to generate 18 structures. the root mean squared deviation (rmsd) of the mean structure is 0.53 a fo ...	1996	8634282
the development of phloem anastomoses between vascular bundles and their role in xylem regeneration after wounding in cucurbita and dahlia.	the differentiation of phloem anastomoses linking the longitudinal vascular bundles has been studied in stem internodes of cucurbita maxima duchesne, c. pepo l. and dahlia pinnata cav. these anastomoses comprise naturally occurring regenerative sieve tubes which redifferentiate from interfascicular parenchyma cells in the young internodes. in all three species, severing a vascular bundle in a young internode resulted in regeneration of xylem to form a curved by-pass immediately around the wound. ...	1996	28321670
improved purification and spectroscopic properties of squash glutamate decarboxylase.	squash glutamate decarboxylase was purified by deae-cellulose batchwise followed by blue-sepharose, cellulofine gcl-2000, and toyopearl hw-55f column chromatography. the purified glutamate decarboxylase had a high specific activity (95.0 u/mg). the absorption spectrum of glutamate decarboxylase had an absorption maximum at 420 nm in the range 300-500 nm. a ph change from 5.3 to 7.8 was accompanied by a decrease in absorbancy at 420 nm. one mole of glutamate decarboxylase contained 3.8 and 1.3 mo ...	1996	27281150
phloem sap proteins from cucurbita maxima and ricinus communis have the capacity to traffic cell to cell through plasmodesmata.	in angiosperms, the functional enucleate sieve tube system of the phloem appears to be maintained by the surrounding companion cells. in this study, we tested the hypothesis that polypeptides present within the phloem sap traffic cell to cell from the companion cells, where they are synthesized, into the sieve tube via plasmodesmata. coinjection of fluorescently labeled dextrans along with size-fractionated cucurbita maxima phloem proteins, ranging in size from 10 to 200 kda, as well as injectio ...	1997	9391168
molecular characterization of a phloem-specific gene encoding the filament protein, phloem protein 1 (pp1), from cucurbita maxima.	sieve elements in the phloem of most angiosperms contain proteinaceous filaments and aggregates called p-protein. in the genus cucurbita, these filaments are composed of two major proteins: pp1, the phloem filament protein, and pp2, the phloem lactin. the gene encoding the phloem filament protein in pumpkin (cucurbita maxima duch.) has been isolated and characterized. nucleotide sequence analysis of the reconstructed gene gpp1 revealed a continuous 2430 bp protein coding sequence, with no intron ...	1997	9263452
effect of disruption of a cutinase gene (cuta) on virulence and tissue specificity of fusarium solani f. sp. cucurbitae race 2 toward cucurbita maxima and c. moschata.	a 3.9-kb genomic dna fragment from the cucurbit pathogen fusarium solani f. sp. cucurbitae race 2 was cloned. sequence analysis revealed an open reading frame of 690 nucleotides interrupted by a single 51-bp intron. the nucleotide and predicted amino acid sequences showed 92 and 98% identity, respectively, to those of the cuta gene of the pea pathogen f. solani f. sp. pisi. a gene replacement vector was constructed and used to generate cuta- mutants that were detected with a polymerase chain rea ...	1997	9100380
gibberellin biosynthesis from gibberellin a12-aldehyde in endosperm and embryos of marah macrocarpus.	soluble enzyme preparations from embryos and endosperm of marah macrocarpus (previously echinocystis macrocarpa) were incubated with [14c4]gibberellin(ga)12-aldehyde, [14c4]ga12, [14c1] ga9, 2,3-didehydro[14c1]ga9, [14c1]ga20, and [17-13c, 3h]ga5. embryo preparations converted ga12-aldehyde, ga12, and ga9 to ga4 and ga7; 2,3-didehydroga9 to ga7; ga5 to ga3; and ga20 (incompletely) to ga1 and ga60, but not to ga3. endosperm preparations converted ga12-aldehyde and ga12 to ga15, ga24, and ga9, but ...	1997	9112781
the ga2 locus of arabidopsis thaliana encodes ent-kaurene synthase of gibberellin biosynthesis.	the ga2 mutant of arabidopsis thaliana is a gibberellin-deficient dwarf. previous biochemical studies have suggested that the ga2 mutant is impaired in the conversion of copalyl diphosphate to ent-kaurene, which is catalyzed by ent-kaurene synthase (ks). overexpression of the previously isolated ks cdna from pumpkin (cucurbita maxima) (cmks) in the ga2 mutant was able to complement the mutant phenotype. a genomic clone coding for ks, atks, was isolated from a. thaliana using cmks cdna as a heter ...	1998	9536043
specificity of human cathepsin g.	a series of tetrapeptide p-nitroanilide substrates of the general formula: suc-ala-ala-pro-aaa-p-nitroanilide was used to map the s1 binding pocket of human cathepsin g. based on the kcat/km parameter, the following order of preference was found: lys=phe>arg=leu>met>nle=nva>ala>asp. thus, the enzyme exhibits clear dual and equal trypsin- and chymotrypsin-like specificities. particularly deleterious were beta-branched side chains of ile and val. the p1 substrate preferences found for cathepsin g ...	1998	9675278
modifications outside the proteinase binding loop in cucurbita maxima trypsin inhibitor iii (cmti-iii) analogues change the binding energy with bovine beta-trypsin.	five 26-peptide analogues of the trypsin inhibitor [pro18]cmti-iii containing leu or tyr in position 7 and val or tyr in position 27: 1 (leu7, tyr27), 2 (tyr7, val27), 3 (tyr7, tyr27), 4 (leu7, val27) and 5 (leu7, ala18, tyr27) were synthesized by the solid-phase method. analogues 1-4 displayed ka with bovine beta-trypsin of the same order of magnitude as the wild cmti-iii inhibitor, whereas for analogue 5, this value was lower by about 3 orders of magnitude. this indicated that for the analogue ...	1998	9781673
ribosome inactivating protein and lectin from bitter melon (momordica charantia) seeds: sequence comparison with related proteins.	one of the ribosome inactivating proteins (rips), beta-momorcharin, and a lectin were isolated from seeds of the bitter melon momordica charantia (family cucurbitaceae) in accordance with published procedures. both beta-momorcharin and m. charantia lectin were then subjected to amino acid sequencing. beta-momorcharin exhibited considerable homology in sequence to other cucurbitaceae rips, and also to the a chains of abrin and ricin which are type ii (double-chained) rips. the resemblance between ...	1998	9875234
the first step of gibberellin biosynthesis in pumpkin is catalyzed by at least two copalyl diphosphate synthases encoded by differentially regulated genes.	the first step in gibberellin biosynthesis is catalyzed by copalyl diphosphate synthase (cps) and ent-kaurene synthase. we have cloned from pumpkin (cucurbita maxima l.) two cdnas, cmcps1 and cmcps2, that each encode a cps. both recombinant fusion cmcps proteins were active in vitro. cps are translocated into plastids and processed by cleavage of transit peptides. for cmcps1 and cmcps2, the putative transit peptides cannot exceed the first 99 and 107 amino acids, respectively, because longer n-t ...	1998	9847116
nmr studies of internal dynamics of serine proteinase protein inhibitors: binding region mobilities of intact and reactive-site hydrolyzed cucurbita maxima trypsin inhibitor (cmti)-iii of the squash family and comparison with those of counterparts of cmti-v of the potato i family.	serine proteinase protein inhibitors follow the standard mechanism of inhibition (laskowski m jr, kato i, 1980, annu rev biochem 49:593-626), whereby an enzyme-catalyzed equilibrium between intact (i) and reactive-site hydrolyzed inhibitor (i) is reached. the hydrolysis constant, khyd, is defined as [i]/[i]. here, we explore the role of internal dynamics in the resynthesis of the scissile bond by comparing the internal mobility data of intact and cleaved inhibitors belonging to two different f ...	1998	9514268
factors improving the accuracy of determination of 15n relaxation parameters in proteins.	a number of factors at all stages of data processing which affect the accuracy of determination of 15n relaxation parameters in 15n-labeled proteins is discussed. methods which allow to improve accuracy of the determined parameters are presented using data obtained for cucurbita maxima trypsin inhibitor.	1999	10698274
phloem long-distance transport of cmnacp mrna: implications for supracellular regulation in plants.	direct support for the concept that rna molecules circulate throughout the plant, via the phloem, is provided through the characterisation of mrna from phloem sap of mature pumpkin (cucurbita maxima) leaves and stems. one of these mrnas, cmnacp, is a member of the nac domain gene family, some of whose members have been shown to be involved in apical meristem development. in situ rt-pcr analysis revealed the presence of cmnacp rna in the companion cell-sieve element complex of leaf, stem and root ...	1999	10498677
analysis of phloem protein patterns from different organs of cucurbita maxima duch. by matrix-assisted laser desorption/ionization time of flight mass spectroscopy combined with sodium dodecyl sulfate polyacrylamide gel electrophoresis.	sieve tubes mediate the long-distance transport of nutrients and signals between source and sink organs of plants. to detect mobile phloem proteins that are differentially distributed in source and sink organs of cucurbita maxima, we used both one-dimensional gel electrophoresis (sds-page) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (maldi-tof ms). both techniques revealed that phloem protein patterns depend on the sampling site: whilst several proteins were ...	1999	10093900
plant paralog to viral movement protein that potentiates transport of mrna into the phloem.	cmpp16 from cucurbita maxima was cloned and the protein was shown to possess properties similar to those of viral movement proteins. cmpp16 messenger rna (mrna) is present in phloem tissue, whereas protein appears confined to sieve elements (se). microinjection and grafting studies revealed that cmpp16 moves from cell to cell, mediates the transport of sense and antisense rna, and moves together with its mrna into the se of scion tissue. cmpp16 possesses the characteristics that are likely requi ...	1999	9872750
translocation of structural p proteins in the phloem.	phloem-specific proteins (p proteins) are particularly useful markers to investigate long-distance trafficking of macromolecules in plants. in this study, genus-specific molecular probes were used in combination with intergeneric grafts to reveal the presence of a pool of translocatable p protein subunits. immunoblot analyses demonstrated that cucurbita spp p proteins pp1 and pp2 are translocated from cucurbita maxima stocks and accumulate in cucumis sativus scions. cucurbita maxima or cucurbita ...	1999	9878637
solution structure by nmr of circulin a: a macrocyclic knotted peptide having anti-hiv activity.	the three-dimensional solution structure of circulin a, a 30 residue polypeptide from the african plant chassalia parvifolia, has been determined using two-dimensional 1h-nmr spectroscopy. circulin a was originally identified based upon its inhibition of the cytopathic effects and replication of the human immunodeficiency virus. structural restraints consisting of 369 interproton distances inferred from nuclear overhauser effects, and 21 backbone dihedral and nine chi1 angle restraints from spin ...	1999	9878410
high-resolution structures of three new trypsin-squash-inhibitor complexes: a detailed comparison with other trypsins and their complexes.	an anionic trypsin from atlantic salmon and bovine trypsin have been complexed with the squash-seed inhibitors, cmti-i (cucurbita maxima trypsin inhibitor i, p1 arg) and cpti-ii (cucurbita pepo trypsin inhibitor ii, p1 lys). the crystal structures of three such complexes have been determined to 1.5-1.8 a resolution and refined to crystallographic r factors ranging from 17.6 to 19.3%. the two anionic salmon-trypsin complexes (st-cpti and st-cmti) and the bovine-trypsin complex (bt-cpti) have been ...	1999	10089404
purification and partial characterization of the pancreatic proteolytic enzymes trypsin, chymotrypsin, and elastase from the chicken.	the purpose of this work was to isolate, purify and partially sequence trypsin, chymotrypsin and elastase from the chicken pancreas. the extraction of the pancreatic zymogens with 0.5 m cacl2 at ph 7.5 for 9 h appeared to be most effective in obtaining maximum recovery of the three enzymes. the sequential cucurbita maxima trypsin inhibitor i/bovine pancreas trypsin inhibitor/soybean trypsin inhibitor affinity chromatography gave the best result for the isolation of trypsin, chymotrypsin and elas ...	1999	10480246
identification and localization of transformed cells in agrobacterium tumefaciens-induced plant tumors	agrobacterium tumefaciens-induced tumors of dicotyledonous plants consist of well-defined vascular bundle-like structures originating from transformed cells. the current view that 25% of the tumor cells are transformed has been re-investigated by using beta-glucuronidase (gus)-gene-containing wild-type bacteria (a281 p35s gus-int). regularly growing stem and leaf tumors showed irregular gus-staining patterns in the different plant species, ricinus communis l., cucurbita maxima l., vicia faba l. ...	1999	10550620
traditional antihelmintic, antiparasitic and repellent uses of plants in central italy.	the uses of 51 plants of marche, abruzzo and latium, distributed in 28 families, are listed here. memories and news of continued use of the plants in these sectors were collected from farmers and shepherds in person (mostly old people). the plants most frequently used as antiparasitics and repellents are juglans regia, lupinus albus, ruta graveolens, fraxinus ornus, datura stramonium, artemisia absinthium, allium cepa, while r. graveolens, cucurbita maxima, a. absinthium, allium sativum are the ...	1999	10624877
design, chemical synthesis and kinetic studies of trypsin chromogenic substrates based on the proteinase binding loop of cucurbita maxima trypsin inhibitor (cmti-iii).	a series of trypsin chromogenic substrates with formula: y-ala-x-abu-pro-lys-pna, where x = gly, ala, abu, val, leu, phe, ser, glu and y = ac, h; pna = p-nitroanilide was synthesized. the cucurbita maxima trypsin inhibitor cmti-iii molecule was used as a vehicle to design the trypsin substrates. to evaluate the influence of position p(4) on the substrate-enzyme interaction, kinetic parameters of newly synthesized substrates with bovine beta-trypsin were determined. the increasing hydrophobicity ...	2000	10694481
yield response of pumpkin and winter squash to simulated cucumber beetle (coleoptera: chrysomelidae) feeding injury.	field studies were conducted to characterize the yield responses of pumpkin, cucurbita pepo l., and wintersquash, cucurbita maxima duch., to simulated striped cucumber beetle, acalymma vittatum (f.), feeding damage. preliminary studies in pumpkin ('spookie') were conducted in 1993, and more complete studies in 1994 and 1995 in pumpkin (spookie), and in 1995 and 1996 in winter squash ('waltham'). plants were artificially injured 1 time at the cotyledon, 1st, 2nd, or 3rd true leaf stages by clippi ...	2000	14658523
proteolytic processing of cmpp36, a protein from the cytochrome b(5) reductase family, is required for entry into the phloem translocation pathway.	cucurbita maxima (pumpkin) phloem sap contains a 31 kda protein that cross-reacts with antibodies directed against the red clover necrotic mosaic virus movement protein (rcnmv mp). microsequence data from phloem-purified 31 kda protein were used to isolate a complementary dna: the open reading frame encodes a 36 kda protein belonging to the cytochrome b(5) reductase (cb5r) family; the gene was termed cmpp36. western analyses established that cmpp36, rcnmv mp and cmpp16 (xoconostle-cázares et al. ...	2000	11135108
electronic structure of trypsin inhibitor from squash seeds in aqueous solution.	the electronic structure of the trypsin inhibitor from seeds of the squash cucurbita maxima (cmti-i) in aqueous solution is obtained by ab initio, all-electron, full-potential calculations using the self-consistent cluster-embedding (scce) method. the reactive site of the inhibitor is explained theoretically, which is in agreement with the experimental results. it is shown that the coordinates of oxygen atoms in the inhibitor, determined by nuclear magnetic resonance and combination of distance ...	2000	11089109
characterization of cucurbita maxima phloem serpin-1 (cmps-1). a developmentally regulated elastase inhibitor.	we report on the molecular, biochemical, and functional characterization of cucurbita maxima phloem serpin-1 (cmps-1), a novel 42-kda serine proteinase inhibitor that is developmentally regulated and has anti-elastase properties. cmps-1 was purified to near homogeneity from c. maxima (pumpkin) phloem exudate and, based on microsequence analysis, the cdna encoding cmps-1 was cloned. the association rate constant (k(a)) of phloem-purified and recombinant his(6)-tagged cmps-1 for elastase was 3.5 + ...	2000	10960478
wound-induced ethylene synthesis and expression and formation of 1-aminocyclopropane-1-carboxylate (acc) synthase, acc oxidase, phenylalanine ammonia-lyase, and peroxidase in wounded mesocarp tissue of cucurbita maxima.	1-aminocyclopropane-1-carboxylate (acc) synthase was rapidly induced in mesocarp tissue of cucurbita maxima after wounding in the cut surface layer in 1 mm thickness (ca. 9 cells) (first layer) in both the enzyme activity and the levels of transcript. this led to a rapid accumulation of acc and hence ethylene production. in the inside tissue (1-2 mm) (second layer), no significant induction of acc synthase was observed, which resulted in a low level of acc, although ethylene was evolved at a muc ...	2000	10845457
matrix-assisted laser desorption/ionization time of flight mass spectrometry peptide mass fingerprints and post source decay: a tool for the identification and analysis of phloem proteins from cucurbita maxima duch. separated by two-dimensional polyacrylamide gel electrophoresis.	a combination of gel electrophoresis and mass spectrometry was used to analyze the soluble proteins from phloem sap of cucurbita maxima duch. phloem proteins were separated using two-dimensional gel electrophoresis. coomassie-stained spots were cut out and subjected to tryptic digestion. to identify proteins, peptide mass fingerprints were determined by matrix-assisted laser desorption/ionization time of flight (maldi-tof) mass spectrometry. in addition, maldi-tof post source decay measurements ...	2001	11556791
disulfide bond effects on protein stability: designed variants of cucurbita maxima trypsin inhibitor-v.	attempts to increase protein stability by insertion of novel disulfide bonds have not always been successful. according to the two current models, cross-links enhance stability mainly through denatured state effects. we have investigated the effects of removal and addition of disulfide cross-links, protein flexibility in the vicinity of a cross-link, and disulfide loop size on the stability of cucurbita maxima trypsin inhibitor-v (cmti-v; 7 kd) by differential scanning calorimetry. cmti-v offers ...	2001	11266603
phloem-specific expression of the pumpkin fruit trypsin inhibitor.	vascular exudates of cucurbita maxima (duch.) contain a group of highly conserved serine proteinase inhibitors collectively called pumpkin fruit trypsin inhibitors (pftis) that prevent proteolytic activity of trypsin or chymotrypsin. polyclonal antibodies raised against pftis were used to immunolocalize these low-molecular-weight proteins within the phloem tissue and to study their developmental expression. the inhibitors were translocated throughout the transport phloem and were present in vasc ...	2001	11216835
induction of cutinolytic esterase activity during saprophytic growth of cucurbit pathogens, fusarium solani f. sp. cucurbitae races one and two (nectria haematococca mpi and mpv, respectively).	cutins from fruit of cucurbita maxima and cucurbita moschata cultivars, apple and a c(16) alcohol (hexadecanol) were used to induce cutinolytic esterase activity during saprophytic growth of strains of the two cucurbit pathogens, fusarium solani f. sp. cucurbitae, race 1 (nectria haematococca mating population (mpi) and f. solani f. sp. cucurbitae, race 2 (mpv). four strains of mpv and 11 strains of mpi were were included in the study. although we were primarily interested in the two cucurbit pa ...	2001	11164297
chromogenic substrates of bovine beta-trypsin: the influence of an amino acid residue in p1 position on their interaction with the enzyme.	the cucurbita maxima trypsin inhibitor cmti-iii molecule was used as a vehicle to design and synthesize a series of trypsin chromogenic substrates modified in position p1: ac-ala-val-abu-pro-x-pna, where x = orn, lys, arg, har, arg(no(2)), cit, hci, phe(p-cn), phe(p-nh(2)); pna = p-nitroanilide. the most active compounds (as determined by specificity constant k(cat)/k(m)) were peptides with the arg and lys residues in the position discussed. changes in the length and the decrease of the positive ...	2001	11478806
applying science to changing dietary patterns.	the intake of 400-600 g/d of fruits and vegetables is associated with reduced incidence of many common forms of cancer. these foods contain phytochemicals that can modulate gene expression to inhibit carcinogenesis via multiple pathways. many phytochemicals are colorful, providing an easy way to communicate increased diversity of fruits and vegetables to the public. red foods contain lycopene, the pigment in tomatoes, which is localized in the prostate gland and may be involved in maintaining pr ...	2001	11694651
a sensitive and rapid bioassay of homogalacturonan synthase using 2-aminobenzamide-labeled oligogalacturonides.	polygalacturonate 4-alpha-galacturonosyltransferase (gala t) activity was detected in the microsomal fraction isolated from pumpkin (cucurbia moschata duchesne, cv. tokyou-kabocha) seedlings using udp-gala and 2-aminobenzamide (2ab)-labeled oligogalacturonides. a 2ab-labeled undecagalacturonide was elongated by the attachment of galacturonic acid (gala) residues to give 2ab-labeled oligogalacturonides with a degree of polymerization (dp) between 12 and 17. exogenous 2ab-labeled oligogalacturonid ...	2002	12461139
a subclass of plant heat shock cognate 70 chaperones carries a motif that facilitates trafficking through plasmodesmata.	plasmodesmata establish a pathway for the trafficking of non-cell-autonomously acting proteins and ribonucleoprotein complexes. plasmodesmal enriched cell fractions and the contents of enucleate sieve elements, in the form of phloem sap, were used to isolate and characterize heat shock cognate 70 (hsc70) chaperones associated with this cell-to-cell transport pathway. three cucurbita maxima hsc70 chaperones were cloned and functional and sequence analysis led to the identification of a previously ...	2002	12456884
relationship between crop losses and initial population densities of meloidogyne arenaria in winter-grown oriental melon in korea.	to determine the economic threshold level, oriental melon (cucumis melo l. cv. geumssaragi-euncheon) grafted on shintozoa (cucurbita maxima x cu. moschata) was planted in plots (2 x 3 m) under a plastic film in february with a range of initial population densities (pi) of meloidogyne arenaria. the relationships of early, late, and total yield to pi measured in september and january were adequately described by both linear regression and the seinhorst damage model. initial nematode densities in s ...	2002	19265907
phylogenetic relationships among domesticated and wild species of cucurbita (cucurbitaceae) inferred from a mitochondrial gene: implications for crop plant evolution and areas of origin.	we have investigated the phylogenetic relationships among six wild and six domesticated taxa of cucurbita using as a marker an intron region from the mitochondrial nad1 gene. our study represents one of the first successful uses of a mtdna gene in resolving inter- and intraspecific taxonomic relationships in angiosperms and yields several important insights into the origins of domesticated cucurbita. first, our data suggest at least six independent domestication events from distinct wild ancesto ...	2002	11782554
distribution of mca-coated grits in maize fields after high wheel tractor application for disrupting orientation of diabrotica virgifera virgifera leconte.	high wheel tractor applications of 4-methoxycinnamaldehyde (mca)-coated corn granules ('grits') were conducted in ruski krstur (serbia) in summer 2001 in a 5 ha corn field. grits are a by-product after corn is harvested and separated from the cob and used as a carrier medium to disseminate mca into the corn field. mca is a kairomone mimic derived form cucurbita maxima (duchesne) used to disrupt orientation of diabrotica virgifera virgifera leconte towards different mca and pheromone baited traps ...	2002	12696417
correlation of binding-loop internal dynamics with stability and function in potato i inhibitor family: relative contributions of arg(50) and arg(52) in cucurbita maxima trypsin inhibitor-v as studied by site-directed mutagenesis and nmr spectroscopy.	the side chains of arg(50) and arg(52) at positions p(6)' and p(8)', respectively, anchor the binding loop to the protein scaffold by means of hydrogen bonds in cucurbita maxima trypsin inhibitor-v (cmti-v), a potato i family member. here, we have investigated the relative contributions of arg(50) and arg(52) to the binding-loop flexibility and stability by determining changes in structure, dynamics, and proteolytic stability as a consequence of individually mutating them into an alanine. we hav ...	2002	12135379
analysis of the complexity of protein kinases within the phloem sieve tube system. characterization of cucurbita maxima calmodulin-like domain protein kinase 1.	in angiosperms, functional, mature sieve elements lack nuclei, vacuoles, ribosomes, and most of the endomembrane network. in this study, the complexity, number, and nature of protein kinases within the phloem sap of cucurbita maxima were investigated to test the hypothesis that the enucleate sieve tube system utilizes a simplified signal transduction network. supporting evidence was obtained in that only five putative protein kinases (three calcium-independent and two calcium-dependent protein k ...	2002	11847230
analysis of highly polar compounds of plant origin: combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry.	the primary goal of metabolomic analysis is the unbiased relative quantification of every metabolite in a biological system. a number of different metabolite-profiling techniques must be combined to make this possible. here we report the separation and analysis of highly polar compounds in a proof of concept study. compounds were separated and analyzed using hydrophilic interaction liquid chromatography (hilic) coupled to electrospray ionization (esi) mass spectrometry. two types of hilic microb ...	2002	11814300
influence of cultivar, cooking, and storage on cell-wall polysaccharide composition of winter squash (cucurbita maxima).	changes in the cell-wall polysaccharides (cwp) of the edible tissues of four winter squash cultivars during storage and after cooking were investigated. a procedure for isolating cell walls of tissues containing high levels of starch was used. the starch-free cwp were sequentially fractionated using cdta, dilute na(2)co(3), and 4 m koh. cellulose made up 40-42% of the total cwp for three cultivars (delica, cf 2, and cf 4) at harvest but was 35% in the softer red warren. the pectic polysaccharide ...	2003	12643650
metabolic networks of cucurbita maxima phloem.	metabolomic analysis aims at a comprehensive characterization of biological samples. yet, biologically meaningful interpretations are often limited by the poor spatial and temporal resolution of the acquired data sets. one way to remedy this is to limit the complexity of the cell types being studied. cucurbita maxima duch. vascular exudates provide an excellent material for metabolomics in this regard. using automated mass spectral deconvolution, over 400 components have been detected in these e ...	2003	12590115
simple and inexpensive flow l-glutamate determination using pumpkin tissue.	this work refers to a very easy to implementate flow injection system with potentiometric detection for l-glutamate determination in food samples. the proposed procedure is based on measurement of carbon dioxide produced by decarboxylation of l-glutamate catalyzed by l-glutamate decarboxylase (e.c. 4.1.1.1.5) from cucurbita maxima (pumpkin). the fi potentiometric system includes an enzymatic reactor with a length of 8 cm and thickness of 5 mm packed with 200 mg of a c. maxima outer layer cut in ...	2003	14611151
modulation of pumpkin glutathione s-transferases by aldehydes and related compounds.	induction of pumpkin (cucurbita maxima duch.) glutathione s-transferase (gst, ec 2.5.1.18) by aldehydes and related compounds was examined. all of the tested compounds induced pumpkin gst to different degrees, and it was found that (1) aldehydes induce gst directly and alcohols induce gst indirectly, and (2) alpha,beta-unsaturated aldehydes are the most effective inducers and their potency is related to the michael acceptors reaction. the results of western blot analysis showed that the patterns ...	2003	12773634
melon resistance to cucurbit yellow stunting disorder virus is characterized by reduced virus accumulation.	abstract the pattern of accumulation of cucurbit yellow stunting disorder virus (cysdv; genus crinivirus, family closteroviridae) rna has been analyzed in several cucurbit accessions. in susceptible accessions of melon (cucumis melo), cucumber (cucumis sativus), marrow (cucurbita maxima), and squash (cucurbita pepo), cysdv rna accumulation peaked during the first to second week postinoculation in the first to third leaf above the inoculated one; younger leaves showed very low or undetectable lev ...	2003	18943165
expression, refolding, and activation of the catalytic domain of human blood coagulation factor xii.	human blood coagulation factor xii (fxii; 80 kda) contains a c-terminal serine protease zymogen domain, which becomes activated upon contacting a negative surface. activated fxii (alphafxiia) brings about reciprocal activation of fxii and kallikrein that by further hydrolysis produces the free catalytic domain (betafxiia; 28 kda). increased levels of alphafxiia are associated with coronary heart disease, sepsis, and diabetes. biophysical investigation of the structural basis of activation, subst ...	2003	12509996
expression studies of gibberellin oxidases in developing pumpkin seeds.	two cdna clones, 3-ox and 2-ox, have been isolated from developing pumpkin (cucurbita maxima) embryos that show significant amino acid homology to gibberellin (ga) 3-oxidases and 2-oxidases, respectively. recombinant fusion protein of clone 3-ox converted ga(12)-aldehyde, ga(12), ga(15), ga(24), ga(25), and ga(9) to ga(14)-aldehyde, ga(14), ga(37), ga(36), ga(13), and ga(4), respectively. recombinant 2-ox protein oxidized ga(9), ga(4), and ga(1) to ga(51), ga(34), and ga(8), respectively. previo ...	2003	12644672
[preclinical studies of cucurbita maxima (pumpkin seeds) a traditional intestinal antiparasitic in rural urban areas].	experimental research was carried out at the parasitology and chemistry laboratories of the jorge basadre grohmann national university, in tacna. the process involved two phases: (1) determination of the minimum inhibitory concentration (mic) of cucurbita maxima as an antiparasitic agent using canine tapeworms with an intestinal isolation of 5 to 6 hours, and (2) determination of the side-effects of curbita maxima on exposed albino rats. it was found that the mic of 23 gr. of pumpkin seed in 100 ...	2004	15614300
proteomics of curcurbit phloem exudate reveals a network of defence proteins.	many different proteins can be separated from the sap of mature sieve tubes of different plant species. to date, only a limited number of those have been identified and functionally characterised. due to sieve tubes inability of transcription and translation, the proteins are most probably synthesised in the intimately connected companion cells and transported into the sieve elements through plasmodesmata. the specific protein composition of phloem sap suggests an important role of these protein ...	2004	15276438
a systemic small rna signaling system in plants.	systemic translocation of rna exerts non-cell-autonomous control over plant development and defense. long-distance delivery of mrna has been proven, but transport of small interfering rna and microrna remains to be demonstrated. analyses performed on phloem sap collected from a range of plants identified populations of small rna species. the dynamic nature of this population was reflected in its response to growth conditions and viral infection. the authenticity of these phloem small rna molecul ...	2004	15258266
xylem sap protein composition is conserved among different plant species.	xylem sap from broccoli (brassica oleracea l. cv. calabrais), rape (brassica napus l. cv. drakkar), pumpkin (cucurbita maxima duch. cv. gelber zentner) and cucumber (cucumis sativus l. cv. hoffmanns giganta) was collected by root pressure exudation from the surface of cut stems of healthy, adult plants. total protein concentrations were in the range of 100 microg ml(-1). one-dimensional gel electrophoresis (sds-page) resulted in 10-20 visible protein bands in a molecular mass range from 10 to 10 ...	2004	15064951
dietary patterns and risk of gastric cancer: a case-control study in uruguay.	gastric cancer is a frequent malignancy in the uruguayan population. in northern counties, incidence rates reach high figures (age-standardized rates [asr], 37.3 per 100,000 men and 18.3 per 100,000 women). diet is a major determinant in gastric carcinogenesis. because foods or food groups have the advantage over nutrients in being most directly related to dietary recommendations, we decided to conduct a case-control study on the relationships between food groups and risk of gastric cancer. for ...	2004	15616769
molecular and biochemical characterization of the capsicum annuum calcium-dependent protein kinase 3 (cacdpk3) gene induced by abiotic and biotic stresses.	the isolated full-length capsicum annuum calcium-dependent protein kinase 3 (cacdpk3) cdna clone was selected from the chili pepper expressed sequence tag database (http://www.pdrc.re.kr/ks200201/pepper.html). phylogenetic analysis based on the deduced amino acid sequence of cacdpk3 cdna revealed significant sequence similarity to the winter squash (cucurbita maxima) cmcpk2 gene (81% identity). genomic gel blot analysis disclosed that cacdpk3 belongs to a multigene family in the pepper genome. c ...	2004	15449060
localization of ascorbic acid, ascorbic acid oxidase, and glutathione in roots of cucurbita maxima l.	to understand the function of ascorbic acid (asc) in root development, the distribution of asc, asc oxidase, and glutathione (gsh) were investigated in cells and tissues of the root apex of cucubita maxima. asc was regularly distributed in the cytosol of almost all root cells, with the exception of quiescent centre (qc) cells. asc also occurred at the surface of the nuclear membrane and correspondingly in the nucleoli. no asc could be observed in vacuoles. asc oxidase was detected by immunolocal ...	2004	15520029
gibberellin biosynthesis in developing pumpkin seedlings.	a gibberellin (ga) biosynthetic pathway was discovered operating in root tips of 7-d-old pumpkin (cucurbita maxima) seedlings. stepwise analysis of ga metabolism in cell-free systems revealed the conversion of ga(12)-aldehyde to bioactive ga(4) and inactive ga(34). highest levels of endogenous ga(4) and ga(34) were found in hypocotyls and root tips of 3-d-old seedlings. cdna molecules encoding two ga oxidases, cmga20ox3 and cmga3ox3, were isolated from root tips of 7-d-old lab150978-treated seed ...	2005	16126862
phenolic profiles of raw apricots, pumpkins, and their purees in the evaluation of apricot nectar and jam authenticity.	the possibility of proving the undeclared addition of pumpkin puree in apricot nectars and jams has been investigated by using the phenol compound fingerprint and sensory evaluation. the cheaper pumpkin admixtures in apricot nectars and jams could not be detected by the sensory evaluation, particularly if present in quantities of <15%. the lower admixtures of pumpkin puree in apricot nectars and jams could be detected by the presence of syringic acid, a phenolic compound characteristic of the in ...	2005	15941324
multi-species interactions impact the accumulation of weathered 2,2-bis (p-chlorophenyl)-1,1-dichloroethylene (p,p'-dde) from soil.	the impact of interactions between the earthworms eisenia foetida and lumbricus terrestris and the plants cucurbita pepo and cucurbita maxima on the uptake of weathered p,p'-dde from soil was determined. although some combinations of earthworm and plant species caused significant changes in the p,p'-dde burden in both organisms, the effects were species specific. contaminant bioconcentration in c. pepo was increased slightly by e. foetida and by 3-fold when the plant was grown with l. terrestris ...	2005	15913856
destination-selective long-distance movement of phloem proteins.	the phloem macromolecular transport system plays a pivotal role in plant growth and development. however, little information is available regarding whether the long-distance trafficking of macromolecules is a controlled process or passive movement. here, we demonstrate the destination-selective long-distance trafficking of phloem proteins. direct introduction, into rice (oryza sativa), of phloem proteins from pumpkin (cucurbita maxima) was used to screen for the capacity of specific proteins to ...	2005	15863519
cucurbit phloem serpins are graft-transmissible and appear to be resistant to turnover in the sieve element-companion cell complex.	serpins are unique inhibitors of serine proteases that are located in various plant tissues and organs. an orthologue of the pumpkin (cucurbita maxima) phloem serpin cmps-1 was amplified from cucumber (cucumis sativus) rna by rt-pcr, cloned, and designated as csps-1 (genbank accession no. aj866989). alternative amino acid sequences in the reactive centre loop suggest distinct inhibitory specificity between cmps-1 and csps-1. a difference in the electrophoretic mobility of these serpins was used ...	2005	16246856
squash xylem sap has activities that inhibit proliferation and promote the elongation of tobacco by-2 cell protoplasts.	to elucidate the physiological functions of the substances in xylem sap, we analyzed the biological activities of xylem sap from squash (cucurbita maxima duch.) root using tobacco by-2 (nicotiana tabacum l. cv. bright yellow 2) cell protoplasts. when by-2 cell protoplasts were cultivated with the total substance of squash xylem sap, the protoplasts elongated remarkably, and cell division was inhibited. although trans-zeatin riboside (zr), the most abundant cytokinin in squash xylem sap, had a co ...	2005	15890518
hispanic and non-hispanic white elders from massachusetts have different patterns of carotenoid intake and plasma concentrations.	carotenoids have been linked with protective roles against diseases associated with aging, including cancer, cardiovascular disease, cataracts, and age-related macular degeneration. with data from a semiquantitative, validated ffq, we examined carotenoid intake of 340 puerto ricans, 98 dominicans, and 146 non-hispanic whites (>60 y old) in massachusetts. compared with non-hispanic white men, hispanic men reported a higher intake of lycopene and lower intakes of alpha-carotene, lutein + zeaxanthi ...	2005	15930459
new analogues of cucurbita maxima trypsin inhibitor iii (cmti iii) with simplified structure.	seven new analogues of trypsin inhibitor cmti iii were obtained by solid-phase peptide synthesis. three analogues contained only two, instead of three, disulfide bridges, whereas the molecules of the next four analogues were shortened at the n- and/or c-terminus. the elimination of one disulfide bridge in cmti iii induces a decrease in the association equilibrium constants by 6-7 orders of magnitude, whereas the removal of one, two or three amino-acid residues at the n- and/or c-terminus does no ...	2006	1418676
a putative acyl-coa-binding protein is a major phloem sap protein in rice (oryza sativa l.).	the n-terminal amino-acid sequence of a major rice phloem-sap protein, named rpp10, was determined. rpp10 is encoded by a single gene in the rice genome. its complete amino-acid sequence, predicted from the corresponding rice full-length cdna, showed high similarity to plant acyl-coa-binding proteins (acbps). western blot analysis using anti-acbp antiserum revealed that putative acbp is abundant in the phloem sap of rice plants, and is also present in sieve-tube exudates of winter squash (cucurb ...	2006	16804052
ectopic expression of pumpkin gibberellin oxidases alters gibberellin biosynthesis and development of transgenic arabidopsis plants.	immature pumpkin (cucurbita maxima) seeds contain gibberellin (ga) oxidases with unique catalytic properties resulting in gas of unknown function for plant growth and development. overexpression of pumpkin ga 7-oxidase (cmga7ox) in arabidopsis (arabidopsis thaliana) resulted in seedlings with elongated roots, taller plants that flower earlier with only a little increase in bioactive ga4 levels compared to control plants. in the same way, overexpression of the pumpkin ga 3-oxidase1 (cmga3ox1) res ...	2006	16384902
organic substances in xylem sap delivered to above-ground organs by the roots.	squash (cucurbita maxima) xylem sap, an apoplastic fluid, contains t-zeatin riboside, glutamine, methylglycine, myo-inositol, fructose, oligosaccharides of arabinogalactan, glucan, galacturonan, and pectins (rhamnogalacturonan-i and rhamnogalacturonan-ii), as well as various proteins, including arabinogalactan and pathogen-related proteins. these substances are mainly produced in stele (xylem) parenchyma and the pericycle in the root-hair zone where ion transporter genes are expressed. glycine-r ...	2006	16733632
the cucurbit images (1515-1518) of the villa farnesina, rome.	the gorgeous frescoes organized by the master renaissance painter raphael sanzio (1483-1520) and illustrating the heavenly adventures of cupid and psyche were painted between 1515 and 1518 to decorate the roman villa (now known as the villa farnesina) of the wealthy sienese banker agostino chigi (1466-1520). surrounding these paintings are festoons of fruits, vegetables and flowers painted by giovanni martini da udine (1487-1564), which include over 170 species of plants. a deconstruction and co ...	2006	16314340