Isolation, purification, and structure of components from acidic polysaccharides of Pleurotus ostreatus (Fr.) Quél.

Isolation of an antitumor component from polysaccharide fraction A5 of some Basidiomyces was achieved by column chromatography on Sephadex G-200. A detection method based on the specific rotatory characteristics of the polysaccharide was applied to estimate components in effluent fractions from the chromatography, and it was confirmed that a series of eluates having similar specific rotation was made up of homogeneous polysaccharide. Three components (H51, H52, and H53) were isolated, in chromatographically pure state, from fraction A5. Component H51 consisted of a skeleton of beta-(1 leads to 3)-linked glucose residues, probably having branches of galactose and mannose residues, and also containing acidic sugars. Component H53 had a main structure similarly consisting of beta-(1 leads to 3)-linked glucose residues and a larger proportion of acidic sugar than H51. Component H52 was a heteropolysaccharide made up of alpha-linked galactose and mannose residues. Components H51 and H53 had a higher and a lower molecular weight, respectively, than H52. The only antitumor-active component was H51.     

Isolation and Identification of Tubaic Acid and β-Tubaic Acid from Derris Roots

A tobacco callus strain, OMT-53, was selected from many cultures as a desirable strain having high nicotine producing capacity. Several culture conditions were examined, aiming to get higher nicotine production with the callus strain, OMT-53. It was revealed that the nicotine production was remarkably enhanced when the callus tissues were cultured at a limited concentration of α-NAA in culture medium. The optimal concentrations of sucrose and nitrogen in the culture medium were 3 % and 840 mg N/L respectively. Some precursors in nicotine biosynthesis were examined, and only ornithine gave a slightly positive effect at 2x10^-4m concentration. Cultures at 25°C produced the highest yield for nicotine. Considerable amounts of nicotine (ca. 20% of total nicotine) were also recognized in the culture medium. Under the best culture condition mentioned above, nicotine production in tobacco callus tissues has been elevated to 2.14% on D.W, basis at 4 weeks’ culture. This value is near to that of the intact tobacco plants.     

Coumarins from the stems and leaves of Clausena dunniana H. Lév

Phytochemical study on the stems and leaves of Clausena dunniana H. Lév. led to the isolation and identification of 14 coumarins (1–14). Their chemical structures were determined on the basis of MS, NMR, and further supported by comparison with those reported in the literature. This is the first report that compounds 1–2, 4, 8, and 12–14 are present in the genus Clausena, and all of these compounds were isolated from C. dunniana for the first time. The chemotaxonomic significance of these isolated compounds was discussed.     

Isolation and Properties of an α-Amylase Inhibitor (0.53) from Wheat (Triticum aestivum)

A droplet gel-entrapping method used for enzyme immobilization was improved to simplify the procedure and to increase the enzyme stability. This immobilization technique is suitable for coupled enzyme reactions requiring cofactors. Leucine dehydrogenase (LeuDH) and formate dehydrogenase (FDH) were freeze-dried with bovine serum albumin, dextrin and stabilizers. The freeze-dried enzyme powder was suspended in a methylcellosolve solution containing polyethyleneglycol(#4000)diacrylate, N,N′-methylenebisacrylamide and 2-hydroxyethylacrylate, and the suspension was gelled with initiators. The gel was cut up and the pieces were washed in a buffer to remove the methylcellosolve and the dextrin inside. The maximum conversion ratio for a LeuDH-FDH gel column was determined to be 99.8% by means of the recycling reaction. On longterm operation at 30 °C for leucine production, the initial conversion ratio (7.2%) gradually decreased to 6.6% over the first 10 days. However, the conversion ratio remained almost constant after the 10th day. The effects of flow rate, temperature, pH, and the concentrations of formate, α- ketoisocaproate, ammonium and NAD on the leucine productivity with the gel column were also investigated.     

Isolation and re-association of the subunits from the pro-(carboxypeptidase A)–pro-(proteinase E) binary complex from pig pancreas

The component subunits of the pro-(carboxypeptidase A)–pro-(proteinase E) binary complex from pig pancreas were separated with a high recovery (80–95%) of their original potential activity. The isolated subunits and the reconstituted complex have properties similar to those of the corresponding natural species. The tryptic activation course of the pro-(carboxypeptidase A) subunit is substantially modified when bound to pro-(proteinase E), whereas the activation of pro-(proteinase E) is not dependent on this association.     

A revision of the moss genus Sciaromium (Mitt.) Mitt. I. General remarks and the section Aloma Dusén

Abstract

The generic name Sciaromium (Mitt.) Mitt. that was originally used as a sectional epithet in Leskea is synonymous with Echinodium Jul E. madeirense Jur. [= E. spinosum (Mitt.) Jur.] is selected as the lectotype of the genus Echinodium. A historical review of taxonomic and nomenclatural problems in Sciaromium is provided and taxa that had previously been excluded from this genus are briefly discussed. Sciaromium flavidulum Dusén is a Eurhynchium and the combination E. flavidulum (Dusén) Ochyra is made. Sciaromium sect. Aloma Dusén is synonymous with Eurhynchium since E. flavidulum serves as the type species of this section. The taxonomic position of S. gracile Dusén is discussed and this species is considered synonymous with Amblystegium chilense Lor., which is transferred to Pseudoleskea and the appropriate new combination P. chilensis (Lor.) Ochyra made. Type specimens of S. bellii Broth., S. elimbatum Broth. & Watts and S. forsythii Broth. in Broth. & Watts are briefly described and illustrated; these species are reduced to synonymy with the polymorphic Cratoneuropsis relaxa (Hook. f. & Wils.) Fleisch. in Broth.     

Isolation and identification of entomopathogenic nematodes and their symbiotic bacteria from Hérault and Gard (Southern France)

Isolation and identification of native nematode-bacterial associations in the field are necessary for successful control of endemic pests in a particular location. No study has yet been undertaken to recover and identify EPN in metropolitan France. In the present paper, we provide results of a survey of EPN and their symbiotic bacteria conducted in Hérault and Gard regions in Southern France. Molecular characterization of isolated nematodes depicted three different Steinernema species and one Heterorhabditis species, H. bacteriophora. Steinernema species recovered were identified as: S. feltiae and S. affine and an undescribed species. Xenorhabdus symbionts were identified as X. bovienii for both S. feltiae and S. affine. Phylogenetic analysis placed the new undescribed Steinernema sp. as closely related to S. arenarium but divergent enough to postulate that it belongs to a new species within the “glaseri-group”. The Xenorhabdus symbiont from this Steinernema sp. was identified as X. kozodoii. All Heterorhabditis isolates recovered were diagnosed as H. bacteriophora and their bacterial symbionts were identified as Photorhabdus luminescens. Molecular characterization of these nematodes enabled the distinction of two different H. bacteriophora strains. Bacterial symbiontic strains of these two H. bacteriophora strains were identified as P. luminescens ssp. kayaii and P. luminescens ssp. laumondii.     

Chemical constituents from the roots of Fallopia convolvulus (L.) A. Löve

Twenty compounds, including three sterols (1–3), three phenols (4, 14 and 15), four anthraquinones (5, 7, 8 and 16), one chromone (6), two stilbenes (9 and 10), three amides (11–13), three flavonoids (17–19) and one organic acid (20), were obtained by modern phytochemical isolation methods. Their structures were identified by spectroscopic methods and in comparison with the published data in the references. Among them, compound 2, 3, 11 and 13 were firstly discovered from genus Fallopia, and compounds 1, 5–8, 10, 14, 15, 17, 19 and 20 were obtained from F. convolvulus for the first time. The chemotaxonomic significance of these compounds was also discussed, which revealed the relationships between F. convolvulus and some other species of Polygonaceae family.     

Protection against herbivores of the myrmecophyte Leonardoxa africana (Baill.) Aubrèv. T3 by its principal ant inhabitant Aphomomyrmex afer Emery

Leonardoxa africana T3 is a myrmecophyte, a plant with specialized structures (domatia) that shelter ants. Adult trees are essentially all occupied by the ant Aphomomyrmex afer. One tree possesses one ant colony. Ants tend homopterans inside the domatia. The plant provides ants with nest sites and food via production of extrafloral nectar and via honeydew produced by homopterans. Workers patrol the young leaves, although their nectaries are not yet functional. This study was conducted to investigate the nature of the relationship between the plant and its ants. In order to determine whether ants protect the plant against herbivorous insects, we placed microlepidopteran larvae on young leaves of several trees, and measured the time until discovery of the larvae by the workers. We then studied the responses of workers as a function of insect size. We showed that workers patrolled the young leaves of the majority of trees. There was, however, inter-colony variability in intensity of patrolling. Workers attacked every larva they found, killing and eating the smaller ones, and chasing larger ones off the young leaf. Most of the phytophagous insects attacking young leaves of L. africana T3 were inventoried in this study. We showed that the larvae of microlepidopterans, one of the most important herbivores of this species, form part of the diet of A. afer. The function of the stereotyped behaviour of ant patrolling on young leaves may be in part to obtain insect protein to complement carbohydrate-rich nectar and honeydew, and in part to protect the host and thus increase its production of resources for ants. Our study shows that ants protect the tree against herbivores, and that even if this protection is less pronounced and more variable than that demonstrated for their sister species L. africana sensu stricto and Petalomyrmex phylax, the association between L. africana T3 and A. afer is a mutualism.     

Isolation and Structural Analysis of an Acidic Polysaccharide from Astragalus membranaceus （Fisch.） Bunge

A new water-soluble hetero-polysaccharlde, APSID3, was obtained from a hot-water extract of the roots of Astragalus membranaceus （Flsch.） Bunge by DEAE-Sepharose Fast Flow and Sephacryl S-300 chromatography. The molecular weight of APSID3 was estimated to be 5.79 × 10^5 Da. Based on a sugar composlUon analysis, methylatlon analysis, partial hydrolysis and 13C nuclear magnetic resonance experimentation, It was concluded that the minimal repeat unit of APSID3 was composed of one terminal arablnose, one 1,5-1Inked arabinose, one 1,3-1Inked rhamnose, one 1,3,4-1Inked rhamnose, five 1,4-1Inked methyl galacturonates and six 1,4-1inked methyl glucuronates.     

Antitumor polysaccharides from P. ostreatus (Fr.) Quél.: isolation and structure of a beta-glucan

We isolated an antitumor glucan (HA beta-glucan) from the neutral polysaccharide fraction (A3) of a hot-water extract of the edible mushroom P. ostreatus (Fr.) Quél. Purification was accomplished by extractions with 20% sodium chloride solution saturated with thymol and by precipitations with ethanol from dimethyl sulfoxide solution. The glucan showed marked antitumor activity at a dose of 0.1 mg/kg. It is a highly branched (1----3)-beta-glucan having an average structure represented by a pentasaccharide segment consisting of one nonreducing terminal, one 3,6-di-O-substituted, and three 3-mono-O-substituted beta-D-glucopyranosyl residues. This structure was confirmed by examining 13C-n.m.r. spectra taken at 75.46 MHz.     

Phenolic and terpenoid compounds from Chione venosa (sw.) urban var. venosa (Bois Bandé)

The Caribbean island of Grenada furnishes the popular aphrodisiac drug Bois Bandé, which consists of the stem bark and the roots of Chione venosa (sw.) URBAN var. venosa (Rubiaceae), a native tree growing in the islands' rain forest. The phytochemical investigation of dichloromethane and methanolic-aqueous extracts of the bark and the roots yielded three acetophenone derivatives described for the first time in plants - ortho-hydroxy-acetophenone-azine (1), acetophenone-2-O-[beta-D-apiofuranosyl-(1'-->6')-O-beta-D-glucopyranoside] (2) and acetophenone-2-O-beta-D-glucopyranoside (3) - along with five known compounds, alpha-morroniside (4), sweroside (5), diderroside (6), daucosterol (7) and beta-sitosterol (8). Their structures were elucidated by 1D and 2D NMR analysis, UV-Vis and ESI-MS.     

Starfish saponins—XI. Isolation and partial characterization of the saponins from the starfish Marthasterias glacialis

• 1.1. The saponin mixture isolated from Marthasterias glacialis was resolved through a series of chromatographic steps into four principal individual components, named marthasteroside A₁, A₂, B and C.
• 2.2. The isolated sulphate steroidal glycosides were characterized by ¹H-NMR, ¹³C-NMR, Fast Atom Bombardment mass spectrometry and GLC analysis of the sugars after acid hydrolysis.
• 3.3. Marthasteroside A₁ and A₂ contained the aglycone thornastrol A and six sugar units. The second group of compounds, marthasteroside B and C, contained five sugar units; the aglycone of marthasteroside B was identified as marthasterone, while that of marthasteroside C was identified as dihydromarthasterone. In all compounds the sulphate group is attached at C-3 of the steroid.

     

Isolation and identification of major ecdysteroids from the pycnogonidPycnogonum litorale (Ström) (Arthropoda,Pantopoda)

Summary From adults ofPycnogonum litorale (Ström) eight ecdysteroids were isolated by HPLC and identified by mass spectrometry and NMR. One of the compounds is 20-hydroxyecdysone, two further ecdysteroids show no OH-group at C-22 (22-deoxy-20,26-dihydroxyecdysone, 22-deoxy-20-hydroxyecdysone=taxisterone). The five other compounds are esters of ecdysteroids with acetic acid (25R and 25S isomers of 20,26-dihydroxyecdysone 22-acetate, 20-hydroxyecdysone 22-acetate) or with glycolic acid (20-hydroxyecdysone 22-glycolate, ecydsone 22-glycolate). The latter are new among zoo- and phytoecdysteroids. No significant amounts of ecdysone could be detected. The origin of the ecdysteroids inPycnogonum litorale and their biological activity are discussed.Abbreviations RP-HPLC Reversed-phase high performance liquid chromatography - NP normal phase - RIA radioimmunoassay - NMR nuclear magnetic resonance - FT Fourier transform - CI/D chemical ionization/desorption - TFA trifluoroacetic acid - E ecdysone - 20E 20-hydroxyecdysone - 2026E 20 26-dihydroxyecdysone     

Studies on sex-organ development. Isolation and characterization of an oestrogen receptor from chick Müllerian duct.

An oestradiol-binding macromolecule was observed in the left Müllerian duct of the 15-day female chick embryo. The embryonic receptor binds oestradiol with a high affinity and low capacity, having a Kd of 3.2 X 10(-9)M and a maximal number of sites of 5.45 fmol/10(6) cells in the left Müllerian duct. The receptor is protein in nature, as suggested by its susceptibility to proteolysis; in addition, it is organ- and steroid-specific. Judging by glycerol-gradient analysis, the hormone receptors in the cytosol are present in 8S and 4.5S forms, and the 8S form could be dissociated into a 4.5S form in the presence of 0.5M-KCl. A 4.5-6S receptor could be extracted from the nuclei. Under physiological salt conditions, the embryonic receptors bind to DNA-cellulose and can be eluted when the salt concentration is increased to 0.5M-KCl. Determination by isoelectric focusing indicates that the isoelectric point is 5.8 for the 8S and 6.9 for the 4.5S receptor.