Production,characteristics and applications of the cell-bound phytase of <Emphasis Type=Italic>Pichia anomala</Emphasis>

Among several yeasts isolated from dried flowers of Woodfordia fruticosa, Pichia anomala produced a high titre of cell-bound phytase. The optimization of fermentation variables led to formulation of media and selection of cultural variables that supported enhanced phytase production. The enzyme productivity was very high in fed batch fermentation in air-lift fermentor as compared to that in stirred tank fermentor. Amelioration in the cell-bound phytase activity was observed when yeast cells were permeabilized with Triton-X-100. The enzyme is thermostable and acid stable with broad substrate specificity, the characteristics that are desirable for enzymes to be used in the animal feed industry. The phytase-encoding gene was cloned and sequenced. The 3D structure of the enzyme was proposed by comparative modeling using phytase of Debaryomyces occidentalis (50% sequence identity) as template. When broiler chicks, and fresh water and marine fishes were fed with the feed supplemented with yeast biomass containing phytase, improvement in growth and phosphorus retention, and decrease in the excretion of phosphorus in the faeces were recorded. The cell-bound phytase of P. anomala could effectively dephytinize wheat flour and soymilk.     

Seed production of cut-leaved teasel (<Emphasis Type=Italic>Dipsacus laciniatus</Emphasis>) in central Missouri

Cut-leaved teasel is an invasive weed in Missouri that reduces the diversification of native species along roadsides and impairs traffic visibility. Teasel is a biennial and grows as a rosette in the first year and flowers the second year. Reproduction is only by seed. Field studies were conducted in 2004 and 2005 at two locations to assess the seed production of cut-leaved teasel. From a natural stand, fifteen plants were tagged at the onset of flowering. Selected plants included those considered growing in a group and those growing alone; a plant was considered alone when no other plant was adjacent for at least 60 cm. Whenever a seedhead completed flowering, it was covered with a cellophane bag and harvested one month later. Linear regression was used to correlate the weight of seeds from a single seedhead and number of seeds to estimate the total seed production per seedhead. The number of seedheads per plant varied from 3 to 56. On average, plants growing alone had 64% more seedheads per plant than plants occurring in a group. Seed numbers in the primary seedhead ranged from 511 to 1,487. Total seed production per plant ranged from 1,309 to 33,527. Seed production was 61% greater for plants growing alone versus those growing in a group and was more prolific in 2005 than in 2004. In addition, seed production per plant varied between locations for plants growing alone, but seed yield per plant was similar for plants growing in groups. Colonization of teasel in new areas is facilitated by higher seedhead numbers per plant and total seed production compared to reproduction of plants in areas of intraspecific competition.     

Tropical Vine Growth and the Effects on Forest Succession: A Review of the Ecology and Management of Tropical Climbing Plants

The climbing habit has evolved independently in many plant taxa, offering vines the ability to compete with non-climbing vegetation for resources such as light, nutrients, and water. This review examines the structural and functional characteristics that allow climbing plants to (1) achieve widespread dispersal, (2) transport large amounts of water throughout vessels, (3) maintain high photosynthesis levels through a large leaf area to biomass ratio, (4) achieve rapid vertical and horizontal expansion by fast growth rates and various climbing mechanisms and (5) survive and recover from disturbances. Due to the competitive effects of vines on trees, management of vine growth is used to preserve tropical timber plantations, combat invasive weeds, and promote rainforest recovery. In order to sustainably manage the vines into the future, it is necessary to understand the mechanisms by which they can alter tropical forest succession and the impacts of various management techniques.     

Palm Harvest Impacts in North-Western South America

Tropical forests harbor thousands of useful plants that are harvested and used in subsistence economies or traded in local, regional or international markets. The effect on the ecosystem is little known, and the forests resilience is badly understood. Palms are the most useful group of plants in tropical American forests. This paper introduces a cross-disciplinary study of the effects of harvesting palm products from the tropical forests in north-western South America. The size of the resource is estimated through palm community studies in the different forest formations that determines the number of species and individuals of all palm species. The genetic structure of useful palm species is studied to determine how much harvesting of the species contributes to genetic erosion of its populations, and whether extraction can be made without harm. Almost all palm species are used in rural communities for subsistence purposes Quantitative ethno-botanical research in different forest types have identified thousands of different ways of using palms for food, construction, tool-making, etc. Although most palms are used by the person harvesting them, many are sold on local markets as fruits, fiber, tools, construction materials etc., and a few have reached larger markets, including international markets. Palm populations are managed in various ways of which some are sustainable and others are destructive. National level mechanisms that governs extraction, trade and commercialization of palm products, are used to identify positive and negative policies in relation to resilience of ecosystems.     

Useful Species Richness,Proportion of Exotic Species,and Market Orientation on Amazonian Dark Earths and Oxisols<Superscript>1</Superscript>

Useful Species Richness, Proportion of Exotic Species, and Market Orientation on Amazonian Dark Earths and Oxisols Anthropogenic soils of Amazonia, known as Amazonian Dark Earths (ADE), are environments with elevated soil fertility that can produce crops that otherwise yield poorly on the leached and highly acidic Oxisols that dominate much of the basin. While ADE sites near urban centers often attract commercial horticultural production of nutrient–demanding exotics, these soils are also considered possibly unique reservoirs of endemic agrobiodiversity because of their relationship to pre–Columbian indigenous occupation. Through botanical surveys and interviews with smallholder farmers, this study compared useful species richness, proportion of exotic species, and market orientation of farms situated on ADE and non–anthropogenic Oxisols in the municipality of Borba in Central Brazilian Amazonia. Species richness was similar on Amazonian Dark Earth and Oxisol farms (19.6 spp vs. 18.3 spp); however, ADE farms showed significantly higher proportions of exotic species (39% vs. 26%; p = 0.025). Furthermore, ADE farms in Borba demonstrated significantly higher market orientation (61.0% vs. 47.3%; p = 0.028), likely a result of the advantage of Amazonian Dark Earths for production of crops that are nutrient–demanding or pH–sensitive crops that have higher values in the nearby regional market of Manaus.     

Parasite-mediated intraguild predation as one of the drivers of co-existence and exclusion among invasive and native amphipods (Crustacea)

Parasitism is emerging as one of the forces determining the outcome of biological invasions. Using field survey and laboratory experiments, we investigate parasitism as one of the factors mediating the interactions among invasive and native amphipods. An extensive survey (100 sites) of a small British island, revealed the native Gammarus duebeni celticus to be parasitised by the muscle wasting microsporidian Pleistophora mulleri and the acanthocephalan duck parasite Polymorphus minutus, the introduced European Gammarus pulex only by P. minutus and the North American Crangonyx pseudogracilis by neither. While Gammarus spp. were widespread in rivers (one or both species present in 64% of sites), C. pseudogracilis had a restricted distribution (7% of sites) and always co-occurred with Gammarus spp. In contrast, Gammarus spp. were absent from all pond/reservoir sites, with C. pseudogracilis present in over 90%. While the negative association of C. pseudogracilis with Gammarus spp. undoubtedly results from factors such as physico-chemical tolerance and predation as C. pseudogracilis can be heavily predated by Gammarus spp., it was notable that C. pseudogracilis co-occurred with Gammarus spp. more frequently when the latter were parasitised. Laboratory experiments clearly showed that predation on C. pseudogracilis was greatly diminished when G. d. celticus was parasitised by P. mulleri and G. pulex by P. minutus. Our study provides evidence that parasitism, by mediating a key interspecific interaction, is one of an array of interacting factors that may have a role in driving patterns of exclusion and co-existence in natives and invaders.     

Inflammation-associated S100 proteins: new mechanisms that regulate function

This review focuses on new aspects of extracellular roles of the calgranulins. S100A8, S100A9 and S100A12 are constitutively expressed in neutrophils and induced in several cell types. The S100A8 and S100A9 genes are regulated by pro- and anti-inflammatory mediators and their functions may depend on cell type, mediators within a particular inflammatory milieu, receptors involved in their recognition and their post-translational modification. The S100A8 gene induction in macrophages is dependent on IL-10 and potentiated by immunosuppressive agents. S100A8 and S100A9 are oxidized by peroxide, hypochlorite and nitric oxide (NO). HOCl generates intra-chain sulfinamide bonds; stronger oxidation promotes cross-linked forms that are seen in human atheroma. S100A8 is >200-fold more sensitive to oxidative cross-linking than low-density lipoprotein and may reduce oxidative damage. S100A8 and S100A9 can be S-nitrosylated. S100A8–SNO suppresses mast cell activation and inflammation in the microcirculation and may act as an NO transporter to regulate vessel tone in inflammatory lesions. S100A12 activates mast cells and is a monocyte and mast cell chemoattractant; a G-protein-coupled mechanism may be involved. Structure–function studies are discussed in relation to conservation and divergence of functions in S100A8. S100A12 induces cytokines in mast cells, but not monocytes/macrophages. It forms complexes with Zn²⁺ and, by chelating Zn²⁺, S100A12 significantly inhibits MMPs. Zn²⁺ in S100A12 complexes co-localize with MMP-9 in foam cells in atheroma. In summary, S100A12 has pro-inflammatory properties that are likely to be stable in an oxidative environment, because it lacks Cys and Met residues. Conversely, S100A8 and S100A9 oxidation and S-nitrosylation may have important protective mechanisms in inflammation.     

Human Alzheimer’s disease synaptic <Emphasis Type=Italic>O</Emphasis>-GlcNAc site mapping and iTRAQ expression proteomics with ion trap mass spectrometry

Neuronal synaptic functional deficits are linked to impaired learning and memory in Alzheimer’s disease (AD). We recently demonstrated that O-GlcNAc, a novel cytosolic and nuclear carbohydrate post-translational modification, is enriched at neuronal synapses and positively regulates synaptic plasticity linked to learning and memory in mice. Reduced levels of O-GlcNAc have been observed in AD, suggesting a possible link to deficits in synaptic plasticity. Using lectin enrichment and mass spectrometry, we mapped several human cortical synaptic O-GlcNAc modification sites. Overlap in patterns of O-GlcNAcation between mouse and human appears to be high, as previously mapped mouse synaptic O-GlcNAc sites in Bassoon, Piccolo, and tubulin polymerization promoting protein p25 were identified in human. Novel O-GlcNAc modification sites were identified on Mek2 and RPN13/ADRM1. Mek2 is a signaling component of the Erk 1/2 pathway involved in synaptic plasticity. RPN13 is a component of the proteasomal degradation pathway. The potential interplay of phosphorylation with mapped O-GlcNAc sites, and possible implication of those sites in synaptic plasticity in normal versus AD states is discussed. iTRAQ is a powerful differential isotopic quantitative approach in proteomics. Pulsed Q dissociation (PQD) is a recently introduced fragmentation strategy that enables detection of low mass iTRAQ reporter ions in ion trap mass spectrometry. We optimized LTQ ion trap settings for PQD-based iTRAQ quantitation and demonstrated its utility in O-GlcNAc site mapping. Using iTRAQ, abnormal synaptic expression levels of several proteins previously implicated in AD pathology were observed in addition to novel changes in synaptic specific protein expression including Synapsin II.     

A novel zinc bis(thiosemicarbazone) complex for live cell imaging

Fluorescent zinc complexes have recently attracted a lot of interest owing to their vast applications in cellular imaging. We report the synthesis as well as physical, chemical and biological studies of a novel zinc glyoxalbis(4-methyl-4-phenyl-3-thiosemicarbazone), [Zn(GTSC)]₃, complex. As compared with the well-studied zinc biacetylbis(4-methyl-3-thiosemicarbazone), Zn(ATSM), complex, which was used as a reference, [Zn(GTSC)]₃ had 2.5-fold higher fluorescence. When cellular fluorescence was measured using flow cytometry, we observed that [Zn(GTSC)]₃ had 3.4-fold to 12-fold higher fluorescence than Zn(ATSM) in various cell lines (n = 9) of different tissue origin. Confocal fluorescence microscopy results showed that [Zn(GTSC)]₃ appeared to have a nuclear localization within 30 min of addition to MCF7 cells. Moreover, [Zn(GTSC)]₃ showed minimal cytotoxicity compared with Zn(ATSM), suggesting that [Zn(GTSC)]₃ may be less deleterious to cells when used as an imaging agent. Our data suggest that the novel [Zn(GTSC)]₃ complex can potentially serve as a biocompatible fluorescent imaging agent for live cells.