Enantioselective Metabolism of Chiral 3-Phenylbutyric Acid, an Intermediate of Linear Alkylbenzene Degradation, by Rhodococcus rhodochrous PB1

Rhodococcus rhodochrous PB1 was isolated from compost soil by selective culture with racemic 3-phenylbutyric acid as the sole carbon and energy source. Growth experiments with the single pure enantiomers as well as with the racemate showed that only one of the two enantiomers, (R)-3-phenylbutyric acid, supported growth of strain PB1. Nevertheless, (S)-3-phenylbutyric acid was cometabolically transformed to, presumably, (S)-3-(2,3-dihydroxyphenyl)butyric acid (the absolute configuration at the C-3 atom is not known yet) by (R)-3-phenylbutyric acid-grown cells of strain PB1, as shown by (sup1)H nuclear magnetic resonance spectroscopy of the partially purified compound and gas chromatography-mass spectrometry analysis of the trimethylsilyl derivative. Oxygen uptake rates suggest that either 3-phenylpropionic acid or cinnamic acid (trans-3-phenyl-2-propenoic acid) is the substrate for aromatic ring hydroxylation. This view is substantiated by the fact that 3-(2,3-dihydroxyphenyl)propionic acid was a substrate for meta cleavage in cell extracts of (R)-3-phenylbutyric acid-grown cells of strain PB1. Gas chromatography-mass spectrometry analysis of trimethylsilane-treated ethyl acetate extracts of incubation mixtures showed that both the meta-cleavage product, 2-hydroxy-6-oxo-2,4-nonadiene-1,9-dicarboxylic acid, and succinate, a hydrolysis product thereof, were formed during such incubations.     

Isoxazolyl, oxazolyl, and thiazolylpropionic acid derivatives as potent alpha(4)beta(1) integrin antagonists

A series of isoxazolyl, oxazolyl, and thiazolylpropionic acid derivatives derived from LDV was found to be a potent antagonist of the alpha(4)beta(1) integrin. The synthesis and SAR leading up to 3-[3-(1-[-[3-methoxy-4-(3-o-tolyl-ureido)-phenyl]-acetylamino]-3-methyl-butyl)-isoxazol-5-yl]-propionic acid (22) are reported. In an allergic mouse model, compound 22 was efficacious delivered systemically (58% inhib @ 10 mg/kg, sc) as well as by intra-tracheal instillation (ED(50)=2 microg/kg).     

Metabolism of Hydroxydibenzofurans, Methoxydibenzofurans, Acetoxydibenzofurans, and Nitrodibenzofurans by Sphingomonas sp. Strain HH69

The metabolism of 11 substituted dibenzofurans by the dibenzofuran-degrading Sphingomonas sp. strain HH69 was investigated. Strain HH69 utilizes 2-, 3-, and 4-acetoxydibenzofuran as well as 2-, 3-, and 4-hydroxydibenzofuran as sole sources of carbon and energy. The degradation of acetoxydibenzofurans is initiated by hydrolysis of the ester bonds, yielding the corresponding hydroxydibenzofurans and acetate. Strain HH69 grew on 2-methoxydibenzofuran only after it was adapted to the utilization of 5-methoxysalicylic acid, whereas 3- and 4-methoxydibenzofuran as well as 2- and 3-nitrodibenzofuran were only cooxidized. During the breakdown of all eight hydroxy-, methoxy-, and nitrodibenzofurans studied here, the corresponding substituted salicylic acids accumulated in the culture broth. In the cases of 2- and 3-hydroxydibenzofuran as well as 2- and 3-nitrodibenzofuran, salicylic acid was also formed. Those four dibenzofurans which did not serve as carbon sources for strain HH69 were converted to a nonutilizable salicylic acid derivative. From turnover experiments with the mutant HH69/II, which is deficient in meta-cleavage, 2,2(prm1),3,4(prm1)-tetrahydroxybiphenyl, 2,2(prm1),3-trihydroxy-5(prm1)-methoxybiphenyl, 2,2(prm1),3-trihydroxy-5(prm1)-nitrobiphenyl, and 2,2(prm1),3-trihydroxy-4(prm1)-nitrobiphenyl were isolated as the main products formed from 3-hydroxydibenzofuran, 2-methoxydibenzofuran, and 2- and 3-nitrodibenzofuran, respectively. These results indicate significant regioselectivity for the dioxygenolytic cleavage of the ether bond of these monosubstituted dibenzofurans, with a preference for the nonsubstituted aromatic nucleus. Substituted trihydroxybiphenyls are converted further by meta-cleavage followed by the removal of the side chain of the resulting product. A stepwise degradation of this side chain was found to be involved in the metabolism of 2-hydroxydibenzofuran.     

Narcissiflorine,narcissiflorinine and narcissifloridine,three triterpene saponins from Anemone narcissiflora

Narcissiflorine, narcissiflorinine and narcissifloridine, three new saponins, have been isolated from the ethanolic extract of Anemone narcissiflora (Ranunculaceae). The structural elucidation of narcissiflorine, narcissiflorinine and narcissifloridine has showed them to be [α-l-arabinofuranosyl-(1 → 4)-β-d-glucuronopyranosyl-(1→3)]- 3β-hydroxy-olean-12-en-28-oic acid, [α,-l-arabinofuranosyl-(1→2)-α-l-rhamnopyranosyl-(1→4)-β-d- glucuronypyranosyl(1→3)]-3-β-hydroxy-olean-12-en-28-oic acid and [α-l-arabinofuranosyl-(1→2)-α-l- rhamnopyranosyl-(1→4)-β-d-glucopyranosyl-(1→3)]-3-β-hydroxy-olean-12-en-28-oic acid, respectively.     

Design, synthesis, and structural analysis of inhibitors of influenza neuraminidase containing a 2,3-disubstituted tetrahydrofuran-5-carboxylic acid core

(+/-)-(2R,3R,5R)-[2-(1'-S-acetamido-3'-methyl)butyl-3-methoxycarbonyl]tetrahydrofuran-5-carboxylic acid (9) and (+/-)-(2R,3R,5R)-[2-(1'-S-acetamido-3'-methyl)butyl-3-(4'-imidazolyl)]tetrahydrofuran 5-carboxylic acid (14) were synthesized as inhibitors of influenza neuraminidase (NA). Both compounds 9 and 14 inhibit influenza NA A with an IC(50) of about 0.5 microM and NA B with an IC(50) of 1.0 microM.     

Derivatives of methanopterin, a coenzyme involved in methanogenesis

Degradational studies of methanopterin, a coenzyme involved in methanogenesis, are reported. The results of these studies are in full accordance with the proposed structure of methanopterin as N-[1'-(2'-amino-4'-hydroxy-7' -methyl-6'-pteridinyl)ethyl]-4-[2', 3', 4', 5'-tetrahydroxypent-1'-yl(5'-1' )O-alpha-ribofuranosyl-5'-phosphoric acid] aniline in which the phosphate group is esterified with alpha-hydroxyglutaric acid. Acid hydrolysis of methanopterin cleaved the 5'----1' glycosidic bond and yielded a 'hydrolytic product' which was identified as N-[1'-(2'-amino-4'-hydroxy-7' -methyl-6'-pteridinyl)ethyl]-4-[2', 3', 4', 5'-tetrahydroxypent-1'-yl]aniline. Alkaline permanganate oxidation of methanopterin yielded 7-methylpterin-6-carboxylic acid. Catalytic (or enzymatic) hydrogenation of methanopterin gave a mixture of 6-ethyl-7-methyl-7,8-dihydropterin, 6-ethyl-7-methylpterin and a third compound, named methaniline which was identified as 4-[2', 3', 4', 5'-tetrahydroxypent-1'-yl(5'----1')O-alpha -ribofuranosyl-5'-phosphoric acid]aniline, in which the phosphate group is esterified with alpha-hydroxyglutaric acid. Methanosarcina barkeri contains a closely related coenzyme called sarcinapterin, which was identified as a L-glutamyl derivative of methanopterin, where the glutamate moiety is attached to the alpha-carboxylic acid group of the alpha-hydroxyglutaric acid moiety of methanopterin via an amide linkage.     

The levels of essential unsaturated fatty acids in human milk on the 3rd, 4th, 5th, and 6th days after labour

The composition of fatty acids in human milk lipids was determined in 41 women on the 3rd, 4th, 5th and 6th days after labour by the method of gas chromatography. In these investigations no significant differences were demonstrated in the fatty acids in the lipid fractions between these consecutive days. The level of polyunsaturated fatty acids of the n-6 and n-3 groups was about 11.9-13.6%, including linoleic acid (18:2, n-6) about 7.7-9.8%, and alpha-linolenic acid (18:3, n-3) about 0.7-1%. In the analysis group of n-6 fatty acids the determined acids were: linoleic acid (18:2, n-6), gamma-linolenic acid (18:3, n-6), eicosadienoic acid (20:2, n-6), eicosatrienoic acid (20:3, n-6), arachidonic acid (20:4, n-6), docosahexaenoic acid (22:6, n-6). From the group of n-3 acids the identified ones were: alpha-linolenic acid (18:3, n-3), eicosapentaenoic acid (20:5, n-3), docosapentaenoic acid (22:5, n-3) and docosahexaenoic acid (22:6, n-3). The obtained quotients of fatty acids n-6 through n-3 on the consecutive days were: 7.2:1-7.8:1, indicating a too low level of the n-3 acids in the investigated milk. The acids prevailing in human milk lipids were: oleic (18:1, n-9) and palmitic (16:0) which accounted for 37-39% and 25-26% respectively. The polyunsaturated to saturated fatty acid ratio (P:S) ranged from 0.28 to 0.33.     

Synthesis, theoretical and structural analyses, and enantiopharmacology of 3-carboxy homologs of AMPA

We have previously used homologation of (S)-glutamic acid (Glu) and Glu analogs as an approach to the design of selective ligands for different subtypes of Glu receptors. (RS)-2-Amino-3-(3-carboxy-5-methyl-4-isoxazolyl)propionic acid (ACPA), which is an isoxazole homolog of Glu, is a very potent agonist at the (RS)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)propionic acid (AMPA) subgroup of Glu receptors and a moderately potent ligand for the kainic acid (KA) subgroup of Glu receptors. The enantiomers of ACPA were previously obtained by chiral HPLC resolution. Prompted by pharmacological interest in ACPA, we have now prepared the (S)- and (R)-enantiomers of ACPA by stereocontrolled syntheses using (1R,2R,5R)- and (1S,2S,5S)-2-hydroxy-3-pinanone, respectively, as chiral auxiliaries. Furthermore, the 5-ethyl analog of ACPA, Ethyl-ACPA, was synthesized, and (S)- and (R)-Ethyl-ACPA were also prepared using this method. The absolute configurations of (S)- and (R)-ACPA were established by X-ray crystallographic analysis of a protected (1S,2S,5S)-2-hydroxy-3-pinanone imine derivative of (R)-ACPA. The absolute stereochemistry of (S)- and (R)-Ethyl-ACPA was assigned on the basis of a comparison of their properties with those of the enantiomers of ACPA, employing elution order on chiral HPLC columns, as well as circular dichroism (CD) spectroscopy in combination with time-dependent density functional theory. The structural and electronic basis for the Cotton effect observed for such analogs is examined. The lower homolog of ACPA, (RS)-2-amino-2-(3-carboxy-5-methyl-4-isoxazolyl)acetic acid (1), which is a Glu analog, was also synthesized. Affinities and neuroexcitatory effects were determined using rat brain membranes and cortical wedges, respectively, at native AMPA, KA, and N-methyl-D-aspartic acid (NMDA) receptors. The molecular pharmacology of (S)- and (R)-ACPA and (S)- and (R)-Ethyl-ACPA was evaluated at homomeric cloned subtypes of AMPA receptors (iGluR1o,3o,4o) and of KA receptors (iGluR5,6), expressed in Xenopus laevis oocytes. The cloned receptors mGluR1alpha, mGluR2, and mGluR4a, expressed in CHO cell lines, were used to study the effects of the five compounds at metabotropic Glu receptors. In accordance with ligand-receptor complexes known from X-ray crystallography, the conformationally restricted Glu analog 1 was inactive at all Glu receptors studied, and the R-forms of ACPA and Ethyl-ACPA were very weak or inactive at these receptors. At AMPA receptor subtypes, (S)-ACPA and (S)-Ethyl-ACPA showed equally potent agonist effects at iGluR1o and iGluR3o, whereas (S)-Ethyl-ACPA was 6-fold more potent than (S)-ACPA at iGluR4o. (S)-ACPA and (S)-Ethyl-ACPA were approximately an order of magnitude less potent at iGluR5 than at AMPA receptor subtypes, and neither compound showed detectable effects at iGluR6. The binding mode of (S)-Ethyl-ACPA at iGluR2 was examined by docking to the (S)-ACPA-iGluR2 complex.     

Resolution, absolute stereochemistry, and enantiopharmacology of the GluR1-4 and GluR5 antagonist 2-amino-3-[5-tert-butyl-3-(phosphonomethoxy)-4-isoxazolyl]propionic acid

The phosphono amino acid, (RS)-2-amino-3-[5-tert-butyl-3-(phosphonomethoxy)-4-isoxazolyl+ ++]propio nic acid (ATPO), is a structural hybrid between the NMDA antagonist (RS)-2-amino-7-phosphonoheptanoic acid (AP7) and the AMPA and GluR5 agonist, (RS)-2-amino-3-(5-tert-butyl-3-hydroxy-4-isoxazolyl)propionic acid (ATPA). ATPO has been resolved into (S)-ATPO and (R)-ATPO using chiral HPLC, and the absolute stereochemistry of the two enantiomers was established by an X-ray crystallographic analysis of (R)-ATPO. (S)-ATPO and (R)-ATPO were characterized pharmacologically using rat brain membrane binding and electrophysiologically using the cortical wedge preparation as well as homo- or heteromeric GluR1-4, GluR5-6, and KA2 receptors expressed in Xenopus oocytes. (R)-ATPO was essentially inactive as an agonist or antagonist in all test systems. (S)-ATPO was an inhibitor of the binding of [(3)H]AMPA (IC(50) = 16 +/- 1 microM) and of [(3)H]-6-cyano-7-nitroquinoxaline-2,3-dione ([(3)H]CNQX) (IC(50) = 1.8 +/- 0.2 microM), but was inactive in the [(3)H]kainic acid and the [(3)H]-(RS)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid ([(3)H]CPP) binding assays. (S)-ATPO did not show detectable agonist effects at any of the receptors under study, but antagonized AMPA-induced depolarization in the cortical wedge preparation (IC(50) = 15 +/- 1 microM). (S)-ATPO also blocked kainic acid agonist effects at GluR1 (K(i) = 2.0 microM), GluR1+2 (K(i) = 3.6 microM), GluR3 (K(i) = 3.6 microM), GluR4 (K(i) = 6.7 microM), and GluR5 (K(i) = 23 microM), but was inactive at GluR6 and GluR6+KA2. Thus, although ATPO is a structural analog of AP7 neither (S)-ATPO nor (R)-ATPO are recognized by NMDA receptor sites.     

Effect of plant growth regulators on ethylene production, 1-aminocyclopropane-1-carboxylic acid oxidase activity, and initiation of inflorescence development of pineapple

With the development of pineapple [Ananas comosus (L.) Merr.] as a fresh fruit crop, it became common to force inflorescence development with ethephon [(2-chloroethyl)phosphonic acid] or ethylene throughout the year. Environmental induction (EI) of inflorescence development disrupts scheduling of fruit harvest and may cause significant losses if small plants are induced, resulting in fruits that are too small to be marketable. Our objective was to identify plant growth regulators (PGRs) that could inhibit EI. Because circumstantial evidence indicates that EI occurs in response to naturally produced ethylene or changes in plant sensitivity to it, most work was done with PGRs that inhibit ethylene biosynthesis or block ethylene action. The synthetic auxin 2-(3-chlorophenoxy)propionic acid (CPA) was included because in one study it reduced the percentage of EI. GA₃, aminooxyacetic acid (AOA), aminoethoxyvinylglycine (AVG), daminozide [butanedioic acid mono-(2,2-dimethylhydrazide)], and silver thiosulfate (STS) had no effect on EL CPA, paclobutrazol [(2RS,3RS)-1-(4-chlorophenyl)methyl-4,4-dimethyl-2(1h-1,2,4-triazol-1-yl)penten-3-ol], and uniconazole [(E)-(p-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol] delayed or inhibited EI of pot-grown pineapple plants. Uniconazole and paclobutrazol inhibited growth and ethylene production by leaf basal-white tissue, and either or both effects could account for the inhibition of EI. Production of 1-aminocyclopropane-1-carboxylic acid (ACC) was unaffected by these compounds, but the activity of ACC oxidase, which converts ACC to ethylene, was inhibited and probably accounts for the reduced ethylene production by leaf basal-white tissue. CPA stimulated ethylene production by stem apical tissue approximately fourfold relative to the control. ACC oxidase activity and the malonyl-ACC (MACC) content in stem apical tissue were also greater than in the control, indicating that CPA greatly stimulated the production of ACC and its sequestration into MACC. The mechanism by which CPA delayed or inhibited EI is not known. CPA, paclobutrazol, and uniconazole appear to have some potential for inhibiting EI of pineapple. Their effect on yield needs to be determined.Abbreviations ACC oxidase 1-aminocyclopropane-1-carboxylic acid oxidase - CPA 2-(3-chlorophenoxy)propionic acid - AOA aminooxyacetic acid - AVG aminoethoxyvinylglycine - daminozide butanedioic acid mono-(2,2-dimethylhydrazide) - DM dry mass - ethephon [(2-chloroethyl)phosphonic acid] - FM fresh mass - GA gibberellin - EI environmental induction of inflorescence development - IA inflorescence appearance - LSD Fisher's protected least significant difference - MACC malonyl-ACC - NAA naphthaleneacetic acid - PGR plant growth regulator - paclobutrazol (2RS,3RS)-1-(4-chlorophenyl)methyl-4,4-dimethyl-2-(1h-1,2,4-triazol-1-yl)penten-3-ol] - uniconazole (E)-(p-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol - STS silver thiosulfate - M-leaf fourth leaf - Ml-L first leaf younger than M-leaf     

Microbial hydroxylation of (+/-)- and (-)-(2Z,4E)-5-(1',2'-epoxy-2',6',6'-trimethylcyclohexyl)-3-methyl-2,4-pentadienoic acid into (+/-)- and (-)-xanthoxin acid by Cunninghamella echinulata

Microbial hydroxylation of (+/-)-(2Z,4E)-5-(1',2'-epoxy-2',6',6'-trimethylcyclohexyl)-3-methyl-2,4-pentadienoic acid (3a) with Cercospora cruenta, a fungus producing (+)-abscisic acid, gave a four-stereoisomeric mixture consisting of (+)- and (-)-xanthoxin acid (4a), and (+)- and (-)-epi-xanthoxin acid (5a) by an HPLC analysis with a chiral column. Screening of the microorganisms capable of oxidizing (+/-)-3a showed that Cunninghamella echinulata stereoselectively oxidized (+/-)-3a to xanthoxin acid (4a) with the some degree of enantioselectivity as (-)-3a to (-)-4a.     

Micropropagation of a fruit tree, Morus australis Poir. syn. M. acidosa Griff.

High frequency bud break and multiple shoots were induced in nodal explants collected between November to February from a 5 year old tree of Morus australis Poir syn. M. acidosa Griff. on Murashige and Skoog's medium supplemented with 6-benzylaminopurine (1.0 mg/1). Incorporation of gibberellic acid (0.3 mg/l) along with BAP (1.0 mg/l) not only induced faster bud break from nodal explants as well as from apical shoot buds, but it also enhanced the frequency of bud break. Nodal explants were more responsive than apical shoot buds. The shoots formed in vitro were multiplied further as nodal segments, and an average multiplication rate of 6-fold per subculture was established within 4–5 months. The shoots were successfully rooted on half-strength MS containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/1. The plantlets were successfully hardened off and established in natural soil.Abbreviations BAP 6-benzylaminopurine - GA₃ gibberellic acid - KN kinetin - IAA indole-3-acetic acid - IBA indole-3-butyric acid - IPA indole-3-propionic acid - MS Murashige and Skoog (1962) medium - NAA 1-naphthalene acetic acid     

Astersedifolioside A-C, three new oleane-type saponins with antiproliferative activity

A phytochemical analysis of Aster sedifolius has led to the isolation of three novel triterpenoid saponins, based on an oleane-type skeleton and named astersedifolioside A (1), B (2) and C (3). On the basis of chemical, and 2D NMR and mass spectrometry data, the structures of the new compounds were elucidated as 3-O-[alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl] echinocystic acid 28-[O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranoside] (1), 3-O-[alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl] echinocystic acid 28-[O-beta-D-xylopyranosyl (1-->4)-O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranoside] (2) and 3-O-[alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl (1-->2)-beta-D-glucopyranosyl] echinocystic acid 28-[O-beta-D-xylopyranosyl (1-->4)-O-alpha-L-rhamnopyranosyl (1-->2)-alpha-L-arabinopyranoside] (3). The isolated compounds showed antiproliferative effect in KiMol, a transformed thyroid cell line.     

Microbiological degradation of bile acids, further degradation of a cholic acid metabolite containing the hexahydroindane nucleus by Corynebacterium equi.

1. The further degradation of a cholic acid (I) metabolite, (4R)-4-[4alpha-(2-carboxyethyl)-3aalpha-hexahydro-7abeta-methyl-5-oxoindan-1beta-yl]valeric acid (IIa), by Corynebacterium equi was investigated. This organism effected ring-opening and gave (4R)-4-[2alpha-(2-carboxyethyl)-3beta-(3-carboxypropionyl)-2beta-methylcyclopent-1beta-yl]valeric acid (VI). The new metabolite was isolated as its trimethyl ester and identified by partical synthesis. It was not utilized by C. equi. 2. (4R)-4[4alpha-(2-Carboxyethyl)-3aalpha-decahydro-8abeta-methyl5-oxa-6-oxoazulen-1beta-yl]valeric acid (IVa), which is a hypothetical initial oxidation product in the above degradation, was not converted by C. equi into the expected metabolite (VI), but into 3 - [2beta - [(2S) - tetrahydro - 5 - oxofur - 2 - yl] - 1beta - methyl - 5 - oxocyclopent - 1alpha - yl]-propionic acid (VIII), the structure of which was established by partial synthesis. 3. Both the possible precursors of the metabolite (VI), an isomer of the epsilon-lactone (IVa), the gamma-lactone (XIa), and the open form of these lactones, the hydroxytricarboxylic acid (V), were also not utilized by C. equi. 4. Under some incubation conditions, C. equi also converted compound (IIa) and 3-(3aalpha-hexahydro-7abeta-methyl-1,5-dioxoindan-4alpha-yl)propionic acid (IIb) into 5-methyl-4-oxo-octane-1,8-dioic acid (III), (4R)-4-(2,3,4,6,6abeta,7,8,9,9aalpha,9bbeta-decahydro-6abeta-methyl-3-oxo-1H-cyclopenta[f]quinolin-7beta-yl)valeric acid (VII) and probably a monohydroxy derivative of compound (IIa) and compound (III), respectively. 5. The possibility that an initial step in the degradation of compound (IIa) by C. equi is oxygenation of the Baeyer-Villiger type, yielding compound (IVa), is discussed. Metabolic pathways of compound (IIa) to compounds (III), (VI), (VII) and (VIII) are also considered.     

In vitro binding affinities of 4-chloro-, 2-methyl-, 4-methyl-, and 4-ethylindoleacetic acid to auxin-binding protein 1 (ABP1) correlate with their growth-stimulating activities

4-Chlorindole-3-acetic acid (4-CI-IAA), an endogenous auxin in certain plant species of Fabaceae, has a higher efficiency in stimulating cell elongation of grass coleoptiles compared with indole-3-acetic acid (IAA), particularly at low concentrations. However, some investigations reported a 1,000-fold discrepancy between growth stimulation and binding affinity of 4-CI-IAA to auxin-binding protein 1 (ABP1) from maize. Here we report binding data of 4-CI-IAA and three alkylated IAA derivatives using purified ABP1 in equilibrium dialysis. There is a clear correlation between the growth-promoting effects and the binding affinity to ABP1 of the different IAA analogues measured by competition of [³H]naphthalene-1-acetic acid binding. Our data are consistent with the hypothesis that ABP1 mediates auxin-induced cell elongation.Abbreviations ABP1 auxin-binding protein 1 - 4-CI-IAA 4-chloroindole-3-acetic acid - NAA naphthalene-1-acetic acid - ER endoplasmic reticulum - IAA indole-3-acetic acid - 2-Me-IAA 2-methylindole-3-acetic acid - 4-Me-IAA 4-methylindole-3-acetic acid - 4-Et-IAA 4-ethylindole-3-acetic acid - MES 4-morpholineethanesulfonic acid - PAA phenylacetic acid     

Effects of dietary polyunsaturated fatty acid deficiencies on mortality, growth and gill structure in the turbot, Scophthalmus maximus

The preparation of fish oil concentrates containing only ( n -3) polyunsaturated fatty acids (PUFA) with different ratios of 20:5 ( n -3)/22:6 ( n -3) is described. Three groups of turbot were maintained on different diets containing: 1, 10% of the dry weight of the diet as natural fish oil, equivalent to 2.5% ( n -3) PUFA and 0–23% ( n -6) PUFA; 2, 10% of the dry weight of the diet as palmitic acid, i.e. no PUFA; 3, 8–7% palmitic acid and 1–3% of the dry weight as ( n -3 PUFA and negligible ( n -6) PUFA. Only the fish on the diet containing natural fish oil showed significant growth over a 15-week period. In addition there were high mortalities on the two experimental diets (2 and 3). Changing the ratio of 20:5 ( n -3)/22:6 ( n -3) from 13–8 to 2–2 in the diet containing 1 3% (n-3) PUFA and negligible ( n -6) PUFA markedly decreased the mortalities. Fish fed the two experimental diets (2 and 3) developed gross changes in gill structure involving the disappearance of chloride cells, a 'sloughing off' of the epithelium along the primary and secondary filaments and an accumulation of cellular material in the inter-lamellar spaces. The tissue ultimately disintegrated to leave a skeleton of connective tissue and a mass of cellular material in the inter-lamellar spaces. It is concluded that 22:6 (n-3) is an essential fatty acid for turbot and that the gill epithelium is a sensitive indicator of this deficiency in this species.     

Synthesis, structure, and conformation of the dilactone derivative of GD1b ganglioside

Treatment of DG1b, beta-Gal-(1----3)-beta-GalNAc-(1---- 4)-[alpha-Neu5Ac-(2----8)-alpha-Neu5Ac-(2---- 3)]-beta-Gal-(1----4)-beta-Glc-(1----1)-Cer, with dicyclohexylcarbodi-imide in anhydrous methyl sulfoxide affords 95-98% of GD1b-dilactone. The carboxyl groups of the two sialic acid units are involved in ester linkages, as proved by ammoniolysis and reduction which gave derivatives containing the amide of sialic acid and N-acetylneuraminulose, respectively. 1H-N.m.r. spectroscopy showed that the lactone rings involved position 9 of the inner sialic acid and position 2 of the inner galactose and that the disialosyl chain is extended toward the -beta-Gal-(1 ----4)-beta-Glc- portion of the ganglioside moiety.     

A comparative study of capacitive immunosensors based on self-assembled monolayers formed from thiourea, thioctic acid, and 3-mercaptopropionic acid

A procedure was developed for the covalent coupling of anti-alpha-fetoprotein antibody (anti-AFP) to a gold surface modified with a self-assembled monolayer (SAM) of thiourea (TU). The performance of the SAM-antibody layer was compared to those of similar layers based on thioctic acid (TA) and 3-mercaptopropionic acid (MPA) by using flow injection capacitive immunosensor system. Covalent coupling of anti-AFP on self-assembled thiourea monolayer (SATUM) modified gold electrode can be used to detect alpha-fetoprotein with high efficiency, similar sensitivity, the same linear range (0.01-10 microgl(-1)) and detection limit (10 ngl(-1)) as those obtained from sensors based on self-assembled thioctic acid monolayer (SATAM) and self-assembled 3-mercaptopropionic acid monolayer (SAMPAM). The system is specific for alpha-fetoprotein and can be regenerated and reused up to 48 times. Therefore, self-assembled monolayer using thiourea which is cheaper than thioctic acid and 3-mercaptopropionic acid is a good alternative for biosensor applications when SAMs are used.     

Bioavailability of Vanadium Extracted by EDTA, HCl, HOAC, and NaNO3 in Topsoil in the Panzhihua Urban Park, Located in Southwest China

Bioavailable vanadium was evaluated on the basis of soil vanadium single-extraction with ethylenediaminetetraacetic acid (EDTA), hydrochloric acid (HCl), acetic acid (HOAc), and sodium nitrate (NaNO(3)) in Panzhihua urban park. The soil vanadium concentration extracted by HOAc was 0.01-2.07?mg?kg(-1), by EDTA 0.28-7.03?mg?kg(-1), by NaNO(3) 0.07-0.53?mg?kg(-1), and by HCl 0.19-1.36?mg?kg(-1). The bioavailable vanadium (bioavailable fraction) obtained with HOAc was 0.01-1.33%, with EDTA 0.27-4.09%, with NaNO(3) 0.13-0.72%, and with HCl 0.06-0.28%. In addition, the impact of soil properties, soil nutrients, and soil enzyme activities on bioavailability of vanadium is discussed in this study. Based on the characteristics of bioavailable vanadium in the soil, ecological and health risks should have been given more attention in the studied area.     

Glycosphingolipids in insects. The amphoteric moiety, N-acetylglucosamine-linked phosphoethanolamine, distinguishes a group of ceramide oligosaccharides from the pupae of Calliphora vicina (Insecta: Diptera)

A group of Calliphora vicina pupal glycolipids could be segregated from the neutral glycosphingolipids, according to their two-dimensional TLC migration properties and positive reactions toward ninhydrin and fluorescamine spray reagents. These classified zwitterionic glycolipids were isolated by silica-gel column chromatography and characterized by the presence of a N-acetyl-glucosamine-bound phosphoethanolamine residue. The structural elucidation of the oligosaccharide moieties was performed by the determination of constituent carbohydrates as alditol acetates, linkage analysis by permethylation, exoglycosidase cleavage, fast-atom-bombardment mass spectrometry and NMR spectroscopy. The dominant fatty acid and sphingoid base species of the ceramide moieties were C20:0 (arachidic acid) and C14:1 (tetradecasphing-4-enine), respectively. The chemical structures of the zwitterionic, biogenetic glycosphingolipid series were determined as: (PEtn-6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer; GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer; GalNAc(alpha 1-4)GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1-3)Man(beta 1- 4)Glc beta Cer; Gal(beta 1-3)GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer; Gal(beta 1-3)GalNAc(alpha 1-4)GalNAc(beta 1-4)(PEtn-6')GlcNAc(beta 1- 3)Man(beta 1-4)Glc beta Cer; GlcNAc(beta 1-3)Gal(beta 1-3)GalNAc(alpha 1-4)GalNAc(beta 1-4)(PEtn- 6')GlcNAc(beta 1-3)Man(beta 1-4)Glc beta Cer.