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PubMed2004

Preparation of alginate/chitosan microcapsules and enteric coated granules of mistletoe lectin.

The aqueous extract of European mistletoe (Viscum album, L.) has been used in cancer therapy. The purified mistletoe lectins, main components of mistletoe, have demonstrated cytotoxic and immune-system-stimulating activities. Korean mistletoe (Viscum album L. coloratum), a subspecies of European mistletoe, has also been reported to possess anticancer and immunological activities. A galactose- and N-acetyl-D-galactosamine-specific lectin (Viscum album L. coloratum agglutinin, VCA) with Mr 60 kDa was isolated from Korean mistletoe. Mistletoe preparations have been given subcutaneously due to the low stability of lectin in the gastrointestinal (GI) tract. In the present study, we investigated the possibility of alginate/chitosan microcapsules as a tool for oral delivery of mistletoe lectin. In addition, our strategy has been to develop a system composed of stabilizing cores (granules), which contain mistletoe lectin, extract or powder, coated by a biodegradable polymer wall. Our results indicated that successful incorporation of VCA into alginate/chitosan microcapsules has been achieved and that the alginate/chitosan microcapsule protected the VCA from degradation at acidic pH values. And coating the VCA with polyacrylic polymers, Eudragit, produced outstanding results with ideal release profiles and only minimal losses of cytotoxicity after manufacturing step. The granules prepared with extract or whole plant produced the best results due to the stability in the extract or whole plant during manufacturing process.

Administration, Oral↗

PubMed2004

Flavonoids of Crotalaria sessiliflora.

Phytochemical investigation of the whole plants of Crotalaria sessiliflora L. led to the isolation of four flavonoids. The structures of these compounds were identified as 2',4',5,7-tetrahydroxyisoflavone (1), 2',4',7-trihydroxyisoflavone (2), 4',7-dihydroxyflavone (3), and isovitexin (4) using spectroscopic analysis. Among these, compounds 2, and 3 have not been reported from Crotalaria species, whereas compounds 1, and 4 were reported from this plant for the first time.

Crotalaria↗

PubMed2003

Ginsenoside Rb1: the anti-ulcer constituent from the head of Panax ginseng.

We previously reported that the butanol (BuOH) fraction of the head of Panax ginseng exhibited gastroprotective activity in peptic and chronic ulcer models. In order to identify the active constituent, an activity-guided isolation of the BuOH faction was conducted with a HCl x ethanol-induced gastric lesion model. The BuOH fraction was passed through a silica-gel column using a chloroform-methanol gradient solvent system, and six fractions (frs. 1-6) were obtained. The active fr. 5 was further separated by silica-gel column, to yield 6 subfractions (subfrs. a-f). Subfr. d was composed of ginsenosides Re, Rc and Rb1. The most active constituent was ginsenoside Rb1 (GRb1), a protopanaxadiol glycoside, which was investigated for its anti-ulcer effect. Gastric injury induced by HCl x ethanol, indomethacin and pyloric ligation (Shay ulcer) was apparently reduced with oral GRb1 doses of 150 and 300 mg/kg. GRb1 at these dosage significantly increased the amount of mucus secretion in an ethanol-induced model. The anti-ulcer effects were consistent with the result of histological examination. These results suggest that the major active constituent in the head of Panax ginseng is GRb1, and that anti-ulcer effect is produced through an increase in mucus secretion.

Animals↗

PubMed2003

A prolyl endopeptidase-inhibiting antioxidant from Phyllanthus ussurensis.

A prolyl endopeptidase inhibitor was isolated from the ethyl acetate soluble fraction of Phyllanthus ussurensis. The active compound was identified as an ellagitannin, corilagin. It was shown to non-competitively inhibit prolyl endopeptidase (PEP) with the IC50 value of 1.17x10(-6) microM. The Ki value was 6.70x10(-7) M. Corilagin was less inhibitory to other serine proteases such as chymotrypsin, trypsin, and elastase, indicating that it was relatively a specific inhibitor of PEP. Corilagin also effectively inhibited reactive oxygen species such as hydroxide and superoxide anion radical, hydrogen peroxide, and DPPH. Especially, corilagin showed potent scavenging activity on the superoxide anion radical in the ESR method (IC50 = 3.79x10(-6) M) as well as xanthine oxidase system.

Antioxidants↗

PubMed2003

Immunomodulatory activity of betulinic acid by producing pro-inflammatory cytokines and activation of macrophages.

Betulinic acid (BA), a pentacyclic triterpene isolated from Lycopus lucidus, has been reported to be a selective inducer of apoptosis in various human cancer and shown anti-inflammatory and immunomodulatory properties. We postulated that BA modulates the immunomodulatory properties at least two groups of protein mediators of inflammation, interleukin-1beta (IL-1beta) and the tumor necrosis factor-alpha (TNF-alpha) on the basis of the critical role of the monocytes and tissue macrophages in inflammatory and immune responses. TNF-alpha and IL-1beta were produced by BA in a dose dependent manner at concentration of 0.625 and 10 microg/mL. The production of NO associated with iNOS was inhibited when treated with LPS at the concentration of 2.5 to 20 microg/mL of BA whereas COX-2 expression was decreased at 2.5 to 20 microg/mL. These modulations of inflammatory mediators were examined in LPS-stimulated RAW 264.7 cells and peritoneal macrophages. The morphology of macrophage was also examined and enhanced surface CD 40 molecule was expressed when treated BA at 0.625 to approximately 5 microg/mL with or without LPS. Furthermore, BA (20 microg/mL) enhanced apoptosis by producing DNA ladder in the RAW 264.7 cells. Our results indicated that BA induced activation of macrophage and pro-inflammatory cytokines. This may provide a molecular basis for the ability of BA to mediate macrophage, suppress inflammation, and modulate the immune response.

Adjuvants, Immunologic↗

PubMed2004

Neuroprotective effect of H. perforatum extracts on beta-amyloid-induced neurotoxicity.

In the present study we assessed the neuroprotective role of a Hypericum perforatum ethanolic extract and obtained fractions in amyloid-beta peptide (Abeta)((25-35))-induced cell death in rat cultured hippocampal neurons. Lipid peroxidation was used as a marker of oxidative stress by following the formation of TBARS in rat cortical synaptosomes, after incubation with ascorbate/Fe2+, alone or in the presence of EC97 effective concentrations of H. perforatum fractions. Induced lipid peroxidation was significantly inhibited by fractions containing flavonol glycosides, flavonol and biflavone aglycones, and by a fraction containing several phenols, mainly chlorogenic acid-type phenolics (21%, 77% and 98%, respectively). Lipid peroxidation evaluated after incubation with 25 microM Abeta(25-35), was significantly inhibited by H. perforatum extract. Cell viability was assessed by use of the Syto-13/PI assay. The total ethanolic extract (TE) and fractions containing flavonol glycosides, flavonol and biflavone aglycones, reduced Abeta(25-35)-induced cell death (65%, 58% and 59%, respectively). These results were further supported by morphological analysis of cells stained with cresyl violet. Peptide beta-amyloid(25-35) induced a decrease in cell volume, chromatin condensation and nuclear fragmentation, alterations not evident in the presence of the TE and fractions containing hypericins (hypericin concentration = 11.02 microM), or fractions containing flavonoids (quercetin concentration = 21.13 microM). Dendritic lesion, an evidence of neurodegeneration, was observed by neuronal staining with cobalt following insult with Abeta(25-35), but prevented after exposure to the peptide plus the fractions referred above. The results of the present paper suggest that H. perforatum extracts may be endowed with neuroprotective compounds able to prevent Abeta(25-35)-induced toxicity.

Amyloid beta-Peptides↗

PubMed2005

Ear of durum wheat under water stress: water relations and photosynthetic metabolism.

The photosynthetic characteristics of the ear and flag leaf of well-watered (WW) and water-stressed (WS) durum wheat (Triticum turgidum L. var. durum) were studied in plants grown under greenhouse and Mediterranean field conditions. Gas exchange measurements simultaneously with modulated chlorophyll fluorescence were used to study the response of the ear and flag leaf to CO2 and O2 during photosynthesis. C4 metabolism was identified by assessing the sensitivity of photosynthetic rate and electron transport to oxygen. The presence of CAM metabolism was assessed by measuring daily patterns of stomatal conductance and net CO2 assimilation. In addition, the histological distribution of Rubisco protein in the ear parts was studied by immunocytochemical localisation. Relative water content (RWC) and osmotic adjustment (osmotic potential at full turgor) were also measured in these organs. Oxygen sensitivity of the assimilation rate and electron transport, the lack of Rubisco compartmentalisation in the mesophyll tissues and the gas-exchange pattern at night indicated that neither C4 nor CAM metabolism occurs in the ear of WW or WS plants. Nevertheless, photosynthetic activity of the flag leaf was more affected by WS conditions than that of the ear, under both growing conditions. The lower sensitivity under water stress of the ear than of the flag leaf was linked to higher RWC and osmotic adjustment in the ear bracts and awns. We demonstrate that the better performance of the ear under water stress (compared to the flag leaf) is not related to C4 or CAM photosynthesis. Rather, drought tolerance of the ear is explained by its higher RWC in drought. Osmotic adjustment and xeromorphic traits of ear parts may be responsible.

Carbon Dioxide↗

PubMed2005

Regulation of the early GA biosynthesis pathway in pea.

The early steps in the gibberellin (GA) biosynthetic pathway are controlled by single copy genes or small gene families. In pea (Pisum sativum L.) there are two ent-kaurenoic acid oxidases, one expressed only in the seeds, while ent-copalyl synthesis and ent-kaurene oxidation appear to be controlled by single copy genes. None of these genes appear to show feedback regulation and the only major developmental regulation appears to be during seed development. During shoot maturation, transcript levels do not change markedly with the result that all the three genes examined are expressed in mature tissue, supporting recent findings that these tissues can synthesise GAs. It therefore appears that the regulation of bioactive GA levels are determined by the enzymes encoded by the 2-oxoglutarate-dependent dioxygenase gene families controlling the later steps in GA biosynthesis. However the early steps are nonetheless important as a clear log/linear relationship exists between elongation and the level of GA1 in a range of single and double mutants in genes controlling these steps.

Alkyl and Aryl Transferases↗

PubMed2004

A genetic analysis of relative growth rate and underlying components in Hordeum spontaneum.

Species from productive and unproductive habitats differ inherently in their relative growth rate (RGR) and a wide range of correlated quantitative traits. We investigated the genetic basis of this trait complex, and specifically assessed whether it is under the control of just one or a few genes that can act as 'master switches' by simultaneously affecting a range of traits in the complex. To address this problem, we crossed two Hordeum spontaneum lines originating from two habitats that differ in productivity. The F3 offspring, in which parental alleles are present in different combinations due to recombination and segregation, was analysed for RGR and its underlying components (leaf area ratio, unit leaf rate, photosynthesis, respiration), as well as a number of other physiological and morphological parameters. For this intra-specific comparison, we found a complex of positively and negatively correlated traits, which was quite similar to what is generally observed across species. A quantitative trait loci (QTL) analysis showed three major and one minor QTL for RGR. Most other variables of the growth-trait complex showed fewer QTLs that were typically scattered over various locations on the genome. Thus, at least in H. spontaneum, we found no evidence for regulation of the trait complex by one or two master switches.

Crosses, Genetic↗

PubMed2006

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-rich protein genes (CRGRPs) cloned by antiserum raised against whole xylem sap of cucumber (Cucumis sativus) were abundantly expressed in the parenchyma cells surrounding xylem vessels in the root-hair zone. CRGRP proteins accumulated and immobilized in the lignified walls of metaxylem vessels and perivascular fibers in shoots, suggesting a systemic delivery mechanism of wall materials via xylem sap. A major 30-kDa protein (XSP30) found in cucumber xylem sap was homologous to the B chains of a lectin (ricin) and bound to a nonfucosylated core N-acetylglucosamine dimer of N-linked glycoproteins abundant in leaf parenchyma cells. XSP30 gene expression, abundant in root xylem parenchyma and pericycle, and the level of XSP30 protein fluctuated diurnally under the control of a circadian clock, and the amplitude was up-regulated by gibberellic acid produced in young leaves, suggesting a long-distance control system between organs.

Cell Wall↗

PubMed2005

Rapid and reliable method of extracting DNA and RNA from sweetpotato, Ipomoea batatas (L). Lam.

A quick, simple and reliable method of extracting DNA from sweetpotato (Ipomoea batatas (L.) Lam.) has been developed. The method was applied successfully for extraction of total DNA from leaves and total RNA from leaves and various tissues. The yield of DNA extracted by this procedure was high (about 1 mg/g leaf tissue). The extracted DNA was completely digested by restriction endonucleases indicating the absence of common contaminating compounds. The absorbancy ratios of A260/A230 and A260/A280 of isolated RNA were approx. 2 and the yield was about 0.2 mg/g fresh wt. CIPK and tublin genes were successfully amplified by RT-PCR, suggesting the integrity of isolated RNA. The total DNA and RNA isolated by this method was of sufficient quality for subsequent molecular analysis.

2-Propanol↗

PubMed2006

GmZFP1 encoding a single zinc finger protein is expressed with enhancement in reproductive organs and late seed development in soybean (Glycine max).

The plant TFIIIA zinc finger proteins play important roles in plant growth and development. In this report, we isolated a single zinc finger gene, designated as GmZFP1, from soybean flowers by in silico mRNA subtraction strategy and RT-PCR. GmZFP1 is an intronless gene and encodes a protein of 210 amino acid residues with a predicted molecular mass of 23.5 KDa. GmZFP1 contains a single zinc finger domain and a DLN-box/EAR-motif at the C-terminus. To localize GmZFP1 mRNA in various tissues, semi-quantative RT-PCR assay was performed. GmZFP1 is expressed in all detected reproductive organs including flowers, developing seeds, pods, sepals, pistils with a low level, and more accumulated in petals, stamens, but not found in all detected vegetative organs with except of stem tips. Further, we found that the expression of GmZFP1 was higher in late seed development than in early seed development. To our knowledge, GmZFP1 is the first characterized gene encoding for single zinc finger protein in soybean, and may play a role as a transcriptional factor in reproductive organs development, especially functioning in petals, stamens and late developing seeds.

Amino Acid Sequence↗

PubMed2005

Activation of the WUS gene induces ectopic initiation of floral meristems on mature stem surface in Arabidopsis thaliana.

A gain-of-function Arabidopsis mutant was identified via activation tagging genetic screening. The mutant exhibited clustered ectopic floral buds on the surface of inflorescence stems. The mutant was designated as sef for stem ectopic flowers. Our detailed studies indicate that the ectopic flower meristems are initiated from the differentiated cortex cells. Inverse PCR and sequence analysis indicated that the enhancer-containing T-DNA from the activation tagging construct, SKI015, was inserted upstream of the previously cloned WUS gene encoding a homeodomain protein. Studies from RT-PCR, RNA in situ hybridization and transgenic plant analysis further confirmed that the phenotypes of sef are caused by the overexpression of WUS. Our results suggest that overexpression of WUS could trigger the cell pluripotence and reestablish a new meristem in cortex. The type of new meristems caused by WUS overexpression was dependent upon the developmental and physiological stages of a plant. With the help of some undefined factors in the reproductive organs the new meristems differentiated into floral buds. In a vegetative growth plant, however, only the new vegetative buds can be initiated upon the overexpression of WUS. These studies provide new insights of WUS on flower development.

Arabidopsis↗

PubMed2005

Use of pruning waste compost as a component in soilless growing media.

The objective of this work was to study the use of pruning wastes compost (PWC) as a growing media component for ornamental plants. The main physical, chemical and biological characteristics of PWC were analysed in order to evaluate its suitability for use in soil-less cultivation. Six growth substrates were prepared by mixing PWC with peat (P), ground leaves (GL), sand (S) and spent mushroom compost (SMC) in different proportions. Two different pot experiments were carried out to test its characteristics of production using perennial ryegrass (Lolium perenne L.) and cypress (Cupressus sempervirens L.) as indicators and the different media as treatments. The growth experiments showed that PWC required mixing with a nutrient-richer material to produce higher results. Therefore, substrates containing SMC (PWC+P+SMC and PWC+SMC) seems to be the most adequate growing media. After the statistical analysis, we concluded that the PWC could be used as a growing media component.

Agaricales↗

PubMed2005

Application of enzymes in producing bleached pulp from wheat straw.

Crude enzymes produced by different strains were used in the production of bleached wheat straw pulp. Pre-treatment with enzymes from Penicillium A10 and Aspergillus L22 at a xylanase dosage of 4 IU/g prior to pulping decreased pulp kappa number by 6.29% and 12.07% respectively as compared to the control. High cellulase activity in crude enzymes has a negative influence on pulping. Xylanase pre-bleaching reduced chlorine charge by 20-30%, or increased final brightness by approximately 4-5% ISO, and improved the pulp strength properties. Xylanase could substitute for alkali extraction in CEH sequence, and be used for treating chemical-bleached pulp, which resulted in higher strength properties for bleached pulp. Modification of bleached pulp with enzymes of 3 IU/g (on xylanase) increased pulp brightness and breaking length by 3-6% ISO and 160-790 m respectively, and decreased post color number and beating degree of pulp by 29-36% and 2.5-5.5 degrees SR respectively, as compared to the original pulp.

Agriculture↗

PubMed2004

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.

Animals↗

PubMed2005

HCHs and DDTs in salt marsh plants (Scirpus) from the Yangtze estuary and nearby coastal areas, China.

HCHs and DDTs in salt marsh plants taken from intertidal flats in the Yangtze estuary and coastal area in April and July 2002 were determined by GC-ECD. A significant seasonal effect was observed for HCHs and DDTs in sources and concentration levels in different sample types including above-ground tissues and roots as well as the whole plants and rhizospheric sediments. The results indicated that the concentration of t-HCH was higher in the above-ground tissues than in their roots in April; however, the partitioning of DDTs between contaminated sediments and the roots showed the higher concentrations of t-DDT in their roots. HCHs and DDTs concentration levels were higher in above-ground tissues than in roots in July. BCFs of HCHs and DDTs exhibited lower values with higher levels of contaminants in sediments, and higher values with lower levels in sediments.

China↗

PubMed2004

Soundwave stimulation triggers the content change of the endogenous hormone of the Chrysanthemum mature callus.

Hormones have been suggested to play a prominent role in the control of callus growth. In this paper, with the method of indirect enzyme-linked immunosorbent assays (ELISA), we investigated the induction effect of soundwave on the dynamic change of endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) in vitro during the differentiation process of Chrysanthemum synchronized mature Callus. These experiments showed that groups treated by optimal soundwave (1.4 kHz, 0.095 kdb) had significantly higher IAA levels and lower ABA than that of the control, which had been implicated activation of endogenous IAA and inhibition of ABA. Through the biochemical analysis, it revealed that the increased level of IAA as well as decreased levels of ABA correlated with soundwave stimulus. High rate of IAA/ABA was favorable to development of the callus and differentiation of mature callus. We conclude that soundwave contributes to endogenous hormone as well as the control of callus growth.

Acoustic Stimulation↗