Intraplastidial lipid trafficking: Regulation of galactolipid release from isolated chloroplast envelope

A method is described for cell-free studies of lipid release from isolated chloroplast envelope. The isolated membrane fraction incorporated radiolabeled galactose into galactolipids, predominantly monogalactosyldiacylglycerol, prior to immobilization of the membrane vesicles onto strips of nitrocellulose. The strips with immobilized membrane were individually incubated with various co-factors and the incubations were terminated by removing the strips. Radioactivity was determined for the strips with immobilized membrane as well as for the material released during the assay. The release of galactolipids from immobilized chloroplast envelope was time- and temperature dependent, required stroma protein(s) and was further stimulated by hydrolysable ATP, GTP and ≤50 μ M acyl-CoAs, of which 16:1-CoA was the most stimulative. To investigate whether guanine nucleotide-binding proteins could be involved, stroma and envelope were independently or together incubated with [ α -³²P]GTP or [ Γ -³²P]GTP. Stroma and envelope proteins were phosphorylated and the envelope fraction contained GMP/GDP binding proteins as well. When the fractions were co-incubated, the patterns of protein phosphorylation and guanine nucleotide binding was different compared to the additive effects of the separate fractions, suggesting that guanine nucleotides may have roles in galactolipid release in addition to providing energy. The results point to several similarities between the regulation of galactolipid release from isolated chloroplast envelope and the regulation of vesicular trafficking among animal and yeast cytosolic membranes, although other mechanisms for lipid release cannot, at this stage, be ruled out.     

Genetic control of arthritis in rats

This study was specifically designed to analyse the genetic control of the chronic disease course for the development of arthritis. Arthritis models with a chronic erosive arthritis are collagen induced arthritis induced with homologous collagen in oil but also arthritis induced with certain non-immunogenic adjuvants such as pristane and avridine. In the presently described experiment we have used pristane induced arthritis. A single injection of 150 μl pristane induces severe chronic arthritis in DA rats. The disease mimics rheumatoid arthritis in many aspects such as the chronic disease course, an erosive inflammation of peripheral joints, symmetric involvement of the joints and the development of rheumatoid factors. To determine the genetic contribution we have used a number of inbred, recombinant inbred and congenic strains as well as specifically designed segregating crosses. An influence by the MHC region (designated Pia1 locus) on the chronic disease course was determined through the uses of MHC congenic LEW strains in which the RT1-f haplotype conferred highest susceptibility. To map genes outside of MHC we used an F2 cross between the highly susceptible DA and the resistant E3 strains. Loci exclusively associated with different phenotypes of the disease could be identified:

Arthritis onset (Pia2 and Pia3).

Severity and joint erosions (Pia4).

Chronicity (Pia5 and Pia6) and Pia1 (determined from MHC congenic (strains)

These findings demonstrates that a chronic self-perpetuative disease, mimicking rheumatoid arthritis, is controlled by different set of genes exclusively linked to different phases of the disease course such as arthritis onset, joint erosions, severity and chronicity.     

On the role of strain in blue copper proteins

Theoretical investigations of the structure and function of the blue copper proteins are described. We have studied the optimum vacuum geometry of oxidised and reduced copper sites, the relative stability of trigonal and tetragonal Cu(II) structures, the relation between the structure and electronic spectra, the reorganisation energy, and reduction potentials. Our calculations give no support to the suggestion that strain plays a significant role in the function of these proteins; on the contrary, our results show that the structures encountered in the proteins are close to their optimal vacuum geometries (within 7 kJ/mol). We stress the importance of defining what is meant by strain and of quantifying strain energies or forces in order to make strain hypotheses testable.     

Mycorrhizal weathering: A true case of mineral plant nutrition?

Weatherable minerals in all podzol surface soils andshallow granitic rock under European coniferousforests studied hitherto are criss-crossed bynumerous open, tubular pores, 3–10 µm in width. Wehypothesize that these pores were formed bycomplex-forming, low-molecular weight organic acidsexuded by or formed in association with mycorrhizalfungi. It is well known that ectomycorrhizal myceliumrepresents a greatly extended, and better distributed,surface area for the absorption of nutrients. However, there have been few investigations of how thewhereabouts of individual hypha affect nutrientuptake. The results presented here provide directevidence that the mycelium is able to penetrate, andmost probably create, microsites which areinaccessible to plant roots and isolated from bulksoil solution phenomena. Dissolved products could betranslocated to the host plant roots, bypassing thesoil solution with often toxic concentration ofAl³⁺ from acid rain, and bypassing competitionfor nutrient uptake by other organisms. Furthermore,there is strong evidence that ``rock-eating'mycorrhizal fungi play a role in the formation ofpodzol E horizons. The partly speculativeinterpretations presented here challenge conventionalideas about (1) the importance of nutrient uptakefrom the bulk soil solution (2) criteria for criticalloads of acid atmospheric deposition for forests, and(3) the process of podzolization.     

Potential Nitrification as an Indicator of Preferential Uptake of Ammonium or Nitrate by Plants in an Oak Woodland Understorey

The preferences of some woodland understorey species for ammoniumand nitrate were investigated by measuring the potential nitrification(conversion of ammonium to nitrate) in the rhizosphere comparedwith the bulk soil. Less acid-tolerant species, which usuallyprefer nitrate or a mixture of ammonium and nitrate in hydroponicculture, should have a higher potential nitrification in therhizosphere compared to the bulk soil due to a low uptake ofammonium (since ammonium is relatively immobile). Acid-tolerantspecies should have a high uptake of ammonium and thereby loweror equal potential nitrification in the rhizosphere comparedto the bulk soil. The hypothesis was tested in a field investigationof five understorey herb species, Deschampsia flexuosa, Convallariamajalis, Poa nemoralis, Geum urbanum andAegopodium podagrariaperformed in oak forests in southern Sweden. Overall, the twoless acid-tolerant species, Geum urbanum and Aegopodium podagraria,had high potential nitrification in the rhizosphere comparedto the bulk soil (indicating a relatively low uptake of ammonium),whilst the acid tolerant species, Deschampsia flexuosa andConvallariamajalis , had approximately equal potential nitrification inthe rhizosphere compared to the bulk soil (indicating a relativelyhigh uptake of ammonium). In the case of Poa nemoralis, a specieswhich grows in both acid and less acid soils, we found the potentialnitrification in the rhizosphere and in the bulk soil to besimilar at low inorganic nitrogen concentrations, but the difference(rhizosphere > bulk) increased when nitrification in thebulk soil was enhanced (i.e. when the nitrogen availabilityincreased). The potential nitrification in the bulk soil variedbetween 0 and 16 nmol g^-1h^-1and was positively correlated withpH. When species occurred at the same site, the potential nitrificationin the bulk soil tended to be lower for the acid tolerant species.Despite a large variation in potential nitrification, the methodoffers a possibility of measuring the preference of plants forammonium/nitrate in a soil system, under natural conditions.Copyright 2000 Annals of Botany Company Ammonium uptake, nitrate uptake, nitrogen preference, potential nitrification, rhizosphere, Deschampsia flexuosa, Convallaria majalis, Poa nemoralis, Geum urbanum, Aegopodium podagraria     

Phylogeny of the tribe Antirrhineae (Scrophulariaceae) based on morphological andndhF sequence data

Phylogenetic relationships within the tribe Antirrhineae (Scrophulariaceae) are analysed and discussed on the basis of parsimony analyses of morphological andndhF gene sequence data. The results indicate that the tribe Antirrhineae consists of four major groups of genera, theAnarrhinum clade, theGambelia clade, theMaurandya clade, and theAntirrhinum clade. TheAnarrhinum clade, consisting of the Old World bee-pollinated generaAnarrhinum andKickxia, is sister to the rest of the tribe. TheGambelia clade consists of the New World generaGambelia andGalvezia, which are very closely related and pollinated by hummingbirds. TheMaurandya clade consists of one subclade includingMaurandya and a number of related bee- or hummingbird-pollinated New World genera and another subclade with the Old World bee-pollinated generaAsarina andCymbalaria. TheAntirrhinum clade consists mainly of bee-pollinated Old World genera, such asAntirrhinum, Linaria, Chaenorhinum, and their segregates, but also includes the New World generaMohavea andHowelliella, of which the latter is known to be partly pollinated by hummingbirds. It is concluded that hummingbirdpollination has evolved independently within Antirrhineae at least three times from bee-pollinated ancestors.     

Effects of different handling systems on pig feed quality

Separation of feed mixtures and its effect on the production traits of growing-finishing pigs was studied in three experiments. In Experiments I and II the effects of different types of equipment to prevent separation in a silo were studied. Experiment III dealt with separation along a feed transport chain.Experiments I and II clearly demonstrated an increase in feed quality variation by filling a silo without any arrangement to reduce separation. The last feed to be extracted from a silo filled in this manner showed an increasing amount of small particles and contents of crude fibre and ash, a decrease in the bulk density and also in the digestibility of nutrients and in the performance of the pigs. Filling a silo equipped with chutes by an auger or pneumatically through a cyclone did not create separation problems, and consequently there were no differences between these treatments in the production experiments.Separation measured by particle size, bulk density or chemical composition was not noticeable along the feed transport chain (Experiment III). There was no effect on performance of the pigs of their location along the chain.     

Assessment of the specific activity of commercial allergen preparations made of house dust in the bacteriosorption reaction: a comparison with the results of the radioallergosorbent inhibition test

The activity of the commercial batches of house-dust (HD) allergens was compared in the inhibition of the radioallergosorbent test (RAST) and in the direct bacteriosorbent test (BST), detecting IgG to the antigens by adsorption on the complexes of whole staphylococcal cells containing protein A. BST was made with rabbit antiserum to HD allergen. This antiserum inhibited RAST by 76% and, therefore, contained antibodies to most of the allergenic determinants of HD. At the same time, no significant correlation between the activity of 15 batches of HD allergen was revealed in RAST inhibition and in BST with the above antiserum. Nevertheless, the exhaustion of the antiserum with a batch of HD allergen showing low activity in RAST inhibition, but high activity in BST made it possible to obtain BST results significantly correlating with the data resulting from RAST inhibition in two series of experiments.     

Discovery of non-peptidergic MrgX1 and MrgX2 receptor agonists and exploration of an initial SAR using solid-phase synthesis

A class of small molecules displaying comparable activities with peptide ligands BAM22 and corticostatin-14 at both the human and rhesus monkey MrgX1 and MrgX2 receptors, respectively, was discovered. A comparative study to compare solid-phase and solution-phase chemistries for the efficient synthesis of the active class, tetracyclic benzimidazoles, was undertaken. The solid-phase chemistry was found to be superior both for the synthesis of analogs and for the synthesis of gram quantities.     

Evolution and loss of long-fringed petals: a case study using a dated phylogeny of the snake gourds, Trichosanthes (Cucurbitaceae)

ABSTRACT: BACKGROUND: The Cucurbitaceae genus Trichosanthes comprises 90-100 species that occur from India to Japan and southeast to Australia and Fiji. Most species have large white or pale yellow petals with conspicuously fringed margins, the fringes sometimes several cm long. Pollination is usually by hawkmoths. Previous molecular data for a small number of species suggested that a monophyletic Trichosanthes might include the Asian genera Gymnopetalum (four species, lacking long petal fringes) and Hodgsonia (two species with petals fringed). Here we test these groups' relationships using a species sampling of c. 60% and 4759 nucleotides of nuclear and plastid DNA. To infer the time and direction of the geographic expansion of the Trichosanthes clade we employ molecular clock dating and statistical biogeographic reconstruction, and we also address the gain or loss of petal fringes. RESULTS: Trichosanthes is monophyletic as long as it includes Gymnopetalum, which itself is polyphyletic. The closest relative of Trichosanthes appears to be the sponge gourds, Luffa, while Hodgsonia is more distantly related. Of six morphology-based sections in Trichosanthes with more than one species, three are supported by the molecular results; two new sections appear warranted. Molecular dating and biogeographic analyses suggest an Oligocene origin of Trichosanthes in Eurasia or East Asia, followed by diversification and spread throughout the Malesian biogeographic region and into the Australian continent. CONCLUSIONS: Long-fringed corollas evolved independently in Hodgsonia and Trichosanthes, followed by two losses in the latter coincident with shifts to other pollinators but not with long-distance dispersal events. Together with the Caribbean Linnaeosicyos, the Madagascan Ampelosicyos and the tropical African Telfairia, these cucurbit lineages represent an ideal system for more detailed studies of the evolution and function of petal fringes in plant-pollinator mutualisms.