Showing papers by "Institut national de la recherche agronomique published in 2000"

Dietary Intake and Bioavailability of Polyphenols

Abstract: The main dietary sources of polyphenols are reviewed, and the daily intake is calculated for a given diet containing some common fruits, vegetables and beverages. Phenolic acids account for about one third of the total intake and flavonoids account for the remaining two thirds. The most abundant flavonoids in the diet are flavanols (catechins plus proanthocyanidins), anthocyanins and their oxidation products. The main polyphenol dietary sources are fruit and beverages (fruit juice, wine, tea, coffee, chocolate and beer) and, to a lesser extent vegetables, dry legumes and cereals. The total intake is approximately 1 g/d. Large uncertainties remain due to the lack of comprehensive data on the content of some of the main polyphenol classes in food. Bioavailability studies in humans are discussed. The maximum concentration in plasma rarely exceeds 1 microM after the consumption of 10-100 mg of a single phenolic compound. However, the total plasma phenol concentration is probably higher due to the presence of metabolites formed in the body's tissues or by the colonic microflora. These metabolites are still largely unknown and not accounted for. Both chemical and biochemical factors that affect the absorption and metabolism of polyphenols are reviewed, with particular emphasis on flavonoid glycosides. A better understanding of these factors is essential to explain the large variations in bioavailability observed among polyphenols and among individuals.

Generalized Discriminant Analysis Using a Kernel Approach

Abstract: We present a new method that we call generalized discriminant analysis (GDA) to deal with nonlinear discriminant analysis using kernel function operator. The underlying theory is close to the support vector machines (SVM) insofar as the GDA method provides a mapping of the input vectors into high-dimensional feature space. In the transformed space, linear properties make it easy to extend and generalize the classical linear discriminant analysis (LDA) to nonlinear discriminant analysis. The formulation is expressed as an eigenvalue problem resolution. Using a different kernel, one can cover a wide class of nonlinearities. For both simulated data and alternate kernels, we give classification results, as well as the shape of the decision function. The results are confirmed using real data to perform seed classification.

Respiration as the main determinant of carbon balance in European forests

Abstract: Carbon exchange between the terrestrial biosphere and the atmosphere is one of the key processes that need to be assessed in the context of the Kyoto Protocol1. Several studies suggest that the terrestrial biosphere is gaining carbon2,3,4,5,6,7,8, but these estimates are obtained primarily by indirect methods, and the factors that control terrestrial carbon exchange, its magnitude and primary locations, are under debate. Here we present data of net ecosystem carbon exchange, collected between 1996 and 1998 from 15 European forests, which confirm that many European forest ecosystems act as carbon sinks. The annual carbon balances range from an uptake of 6.6 tonnes of carbon per hectare per year to a release of nearly 1 t C ha-1 yr-1, with a large variability between forests. The data show a significant increase of carbon uptake with decreasing latitude, whereas the gross primary production seems to be largely independent of latitude. Our observations indicate that, in general, ecosystem respiration determines net ecosystem carbon exchange. Also, for an accurate assessment of the carbon balance in a particular forest ecosystem, remote sensing of the normalized difference vegetation index or estimates based on forest inventories may not be sufficient.

Proanthocyanidins and tannin-like compounds – nature, occurrence, dietary intake and effects on nutrition and health

Abstract: Proanthocyanidins (syn condensed tannins) are complex flavonoid polymers naturally present in cereals, legume seeds and particularly abundant in some fruits and fruit juices. They share some common structural features—phenolic nature and high molecular weight—with phenolic polymers found in black tea and red wine (called here tannin-like compounds). The polymeric nature of proanthocyanidins makes their analysis and estimation in food difficult. For this reason, little is known about their consumption, although they likely contribute a large part of the daily polyphenol intake. They also share common physicochemical properties: they form stable complexes with metal ions and with proteins and are, like other polyphenols, good reducing agents. Many of their biological effects of nutritional interest derive from these properties. As metal ion chelators, they influence the bioavailability of several minerals. The nutritional significance of the non-specific complexation of proteins is less clear. As reducing agents, they may participate in the prevention of cancers, both of the digestive tract and inner organs. They may also protect LDLs against oxidation and inhibit platelet aggregation and therefore prevent cardiovascular diseases. In vitro, animal and human studies on the prevention of these chronic diseases are reviewed with particular attention to wine and tea polyphenols. The lack of data on their bioavailability and the paucity of human studies are emphasised. © 2000 Society of Chemical Industry

Methane production by ruminants: its contribution to global warming

Abstract: The aim of this paper is to review the role of methane in the global warming scenario and to examine the contribution to atmospheric methane made by enteric fermentation, mainly by ruminants. Agricultural emissions of methane in the EU-15 have recently been estimated at 10.2 million tonnes per year and represent the greatest source. Of these, approximately two-thirds come from enteric fermentation and one-third from livestock manure. Fermentation of feeds in the rumen is the largest source of methane from enteric fermentation and this paper considers in detail the reasons for, and the consequences of, the fact that the molar percentage of the different volatile fatty acids produced during fermentation influences the production of methane in the rumen. Acetate and butyrate promote methane production while propionate formation can be considered as a competitive pathway for hydrogen use in the rumen. The many alternative approaches to reducing methane are considered, both in terms of reduction per animal and reduction per unit of animal product. It was concluded that the most promising areas for future research for reducing methanogenesis are the development of new products/delivery systems for anti-methanogenic compounds or alternative electron acceptors in the rumen and reduction in protozoal numbers in the rumen. It is also stressed that the reason ruminants are so important to mankind is that much of the world's biomass is rich in fibre. They can convert this into high quality protein sources (i.e. meat and milk) for human consumption and this will need to be balanced against the concomitant production of methane.

Disruption of the uncoupling protein-2 gene in mice reveals a role in immunity and reactive oxygen species production.

Abstract: The gene Ucp2 is a member of a family of genes found in animals and plants, encoding a protein homologous to the brown fat uncoupling protein Ucp1 (refs 1–3). As Ucp2 is widely expressed in mammalian tissues4,5, uncouples respiration6 and resides within a region of genetic linkage to obesity4, a role in energy dissipation has been proposed. We demonstrate here, however, that mice lacking Ucp2 following targeted gene disruption are not obese and have a normal response to cold exposure or high-fat diet. Expression of Ucp2 is robust in spleen, lung and isolated macrophages4,5,7, suggesting a role for Ucp2 in immunity or inflammatory responsiveness4. We investigated the response to infection with Toxoplasma gondii in Ucp2−/− mice, and found that they are completely resistant to infection, in contrast with the lethality observed in wild-type littermates. Parasitic cysts and inflammation sites in brain were significantly reduced in Ucp2−/− mice (63% decrease, P<0.04). Macrophages from Ucp2 −/− mice generated more reactive oxygen species than wild-type mice (80% increase, P<0.001) in response to T. gondii, and had a fivefold greater toxoplasmacidal activity in vitro compared with wild-type mice (P<0.001 ), which was absent in the presence of a quencher of reactive oxygen species (ROS). Our results indicate a role for Ucp2 in the limitation of ROS and macrophage-mediated immunity.

Relationship of soil organic matter dynamics to physical protection and tillage

Abstract: Tillage has been reported to reduce organic matter concentrations and increase organic matter turnover rates to a variable extent. The change of soil climate and the incorporation of aboveground C inputs within the soil lead to no unique effect on biodegradation rates, because of their strong interaction with the regional climate and the soil physical properties. The periodical perturbation of soil structure by tools and the subsequent drying‐rewetting cycles may be the major factor increasing organic matter decomposition rates by exposing the organic matter that is physically protected in microaggregates to biodegradation. This paper reviews the assessed effects of tillage on organic matter, the scale, extent and mechanisms of physical protection of organic matter in soils. # 2000 Elsevier Science B.V. All rights reserved.

Cytochromes P450: a success story

Abstract: Cytochrome P450 proteins, named for the absorption band at 450 nm of their carbon-monoxide-bound form, are one of the largest superfamilies of enzyme proteins. The P450 genes (also called CYP) are found in the genomes of virtually all organisms, but their number has exploded in plants. Their amino-acid sequences are extremely diverse, with levels of identity as low as 16% in some cases, but their structural fold has remained the same throughout evolution. P450s are heme-thiolate proteins; their most conserved structural features are related to heme binding and common catalytic properties, the major feature being a completely conserved cysteine serving as fifth (axial) ligand to the heme iron. Canonical P450s use electrons from NAD(P)H to catalyze activation of molecular oxygen, leading to regiospecific and stereospecific oxidative attack of a plethora of substrates. The reactions carried out by P450s, though often hydroxylation, can be extremely diverse and sometimes surprising. They contribute to vital processes such as carbon source assimilation, biosynthesis of hormones and of structural components of living organisms, and also carcinogenesis and degradation of xenobiotics. In plants, chemical defense seems to be a major reason for P450 diversification. In prokaryotes, P450s are soluble proteins. In eukaryotes, they are usually bound to the endoplasmic reticulum or inner mitochondrial membranes. The electron carrier proteins used for conveying reducing equivalents from NAD(P)H differ with subcellular localization. P450 enzymes catalyze many reactions that are important in drug metabolism or that have practical applications in industry; their economic impact is therefore considerable.

The genome sequence of the plant pathogen Xylella fastidiosa

Abstract: Instituto Ludwig de Pesquisa sobre o Câncer, Rua Prof. Antonio Prudente, 109-4 andar, 01509-010, Sao Paulo-SP

Membrane proteins and proteomics: un amour impossible?

Abstract: Proteome analysis implies the ability to separate proteins as a first step prior to characterization. Thus, the overall performance of the analysis strongly depends on the performance of the separation tool, usually two-dimensional electrophoresis. This review shows how two-dimensional electrophoresis performs with membrane proteins from bacteria or animal or vegetable cells and tissues, the recent progress in this field, and it examines future prospects in this area.

A Mutation in PRKAG3 Associated with Excess Glycogen Content in Pig Skeletal Muscle

Abstract: A high proportion of purebred Hampshire pigs carries the dominant RN- mutation, which causes high glycogen content in skeletal muscle. The mutation has beneficial effects on meat content but detrimental effects on processing yield. Here, it is shown that the mutation is a nonconservative substitution (R200Q) in the PRKAG3 gene, which encodes a muscle-specific isoform of the regulatory gamma subunit of adenosine monophosphate-activated protein kinase (AMPK). Loss-of-function mutations in the homologous gene in yeast (SNF4) cause defects in glucose metabolism, including glycogen storage. Further analysis of the PRKAG3 signaling pathway may provide insights into muscle physiology as well as the pathogenesis of noninsulin-dependent diabetes mellitus in humans, a metabolic disorder associated with impaired glycogen synthesis.

Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector

Abstract: We have developed a system for stable germline transformation in the silkworm Bombyx mori L. using piggyBac, a transposon discovered in the lepidopteran Trichoplusia ni. The transformation constructs consist of the piggyBac inverted terminal repeats flanking a fusion of the B. mori cytoplasmic actin gene BmA3 promoter and the green fluorescent protein (GFP). A nonautonomous helper plasmid encodes the piggyBac transposase. The reporter gene construct was coinjected into preblastoderm eggs of two strains of B. mori. Approximately 2% of the individuals in the G1 broods expressed GFP. DNA analyses of GFP-positive G1 silkworms revealed that multiple independent insertions occurred frequently. The transgene was stably transferred to the next generation through normal Mendelian inheritance. The presence of the inverted terminal repeats of piggyBac and the characteristic TTAA sequence at the borders of all the analyzed inserts confirmed that transformation resulted from precise transposition events. This efficient method of stable gene transfer in a lepidopteran insect opens the way for promising basic research and biotechnological applications.

Ruminant milk fat plasticity: nutritional control of saturated, polyunsaturated, trans and conjugated fatty acids

Abstract: After a brief survey of metabolic pathways and nutrient fluxes involved in mammary lipogenesis, this review summarises the known effects of diet on ruminant milk fat composition. Special attention is given to fatty acids that could play a positive role for human health, such as butyric acid, oleic acid, C18 to C22 polyunsaturated fatty acids and conjugated linoleic acid (CLA). The efficiency of the transfer of C18:2, C18:3, C20:5, C22:5 and C22:6, from the duodenum to the milk, is reviewed. The main dietary factors taken into account are the nature of forages, including pasture, and the supplementation of dairy rations with protected or unprotected vegetable or fish oils. Dose-response curves of milk CLA are reviewed for different fat supplements, as well as the non-linear relationship between milk CLA and trans C18:1. The potential of dietary factors to increase the mean CLA content in cow milk fat is about 300% above basal values. There is, however, a need to evaluate how the different feeding strategies could change the other aspects of milk fat quality.

The TT8 Gene Encodes a Basic Helix-Loop-Helix Domain Protein Required for Expression of DFR and BAN Genes in Arabidopsis Siliques

Abstract: The TRANSPARENT TESTA8 (TT8) locus is involved in the regulation of flavonoid biosynthesis in Arabidopsis. The tt8-3 allele was isolated from a T-DNA–mutagenized Arabidopsis collection and found to be tagged by an integrative molecule, thus permitting the cloning and sequencing of the TT8 gene. TT8 identity was confirmed by complementation of tt8-3 and sequence analysis of an additional allele. The TT8 gene encodes a protein that displays a basic helix-loop-helix at its C terminus and represents an Arabidopsis ortholog of the maize R transcription factors. The TT8 transcript is present in developing siliques and in young seedlings. The TT8 protein is required for normal expression of two flavonoid late biosynthetic genes, namely, DIHYDROFLAVONOL 4-REDUCTASE (DFR) and BANYULS (BAN), in Arabidopsis siliques. Interestingly, TRANSPARENT TESTA GLABRA1 (TTG1) and TT2 genes also control the expression of DFR and BAN genes. Our results suggest that the TT8, TTG1, and TT2 proteins may interact to control flavonoid metabolism in the Arabidopsis seed coat.

Organic Matter Influence on Clay Wettability and Soil Aggregate Stability

Abstract: Soil organic matter is thought to increase aggregate stability by lowering the wettability and increasing the cohesion of aggregates. In southwest France, thick humic loamy soils (Vermic Haplubrepts) have been intensively cropped for 40 yr, decreasing the soil organic pool and lowering the soil agregate stability. This study assessed (i) the contribution of organic matter to aggregate stability by decreasing aggregate wettability and (ii) the specific role of clay-associated organic matter. Soil samples with a C content of 4 to 53 g kg -1 were sampled and soil aggregate stability was measured. Aggregate wettability was assessed by measuring water drop penetration times on individual 3-to 5-mm aggregates. The <2-μm fractions were extracted without organic matter destruction and their wettability was determined by measuring contact angles of water on clay deposits. Aggregate stability against slaking was correlated to soil C content (r 2 = 0.71 for fast wetting). Water drop penetration time increased with C contents from 1 to 32 s and was very heterogeneous among individual aggregates from a given soil. The contact angle of water on the clay fraction increased linearly with the C content (r 2 = 0.86). This change in clay wettability could partly explain the higher water stability of soils rich in C.

Protein–polysaccharide interactions

Abstract: Numerous investigations on protein–polysaccharide systems have recently been undertaken and are leading to a better understanding of the key parameters implied in protein–polysaccharide interactions. Microscopic methods are being developed to describe the structure formation in the mixed systems in combination with rheological characterisation. Progress is also being made in the description of the mechanisms underlying the phase separation processes by the use of scattering techniques.

Sulforaphane, a Naturally Occurring Isothiocyanate, Induces Cell Cycle Arrest and Apoptosis in HT29 Human Colon Cancer Cells

Abstract: Sulforaphane is an isothiocyanate that is present naturally in widely consumed vegetables and has a particularly high concentration in broccoli. This compound has been shown to block the formation of tumors initiated by chemicals in the rat. Although sulforaphane has been proposed to modulate the metabolism of carcinogens, its mechanism of action remains poorly understood. We have previously demonstrated that sulforaphane inhibits the reinitiation of growth and decreases the cellular viability of quiescent human colon carcinoma cells (HT29). Moreover, the weak effect observed on differentiated CaCo2 cells suggests a specific anticancer activity for this compound. Here we investigated the effect of sulforaphane on the growth and viability of HT29 cells during their exponentially growing phase. We observed that sulforaphane induced a cell cycle arrest in a dose-dependent manner, followed by cell death. This sulforaphane-induced cell cycle arrest was correlated with an increased expression of cyclins A and B1. Moreover, we clearly demonstrated that sulforaphane induced cell death via an apoptotic process. Indeed, a large proportion of treated cells display the following: (a) translocation of phosphatidylserine from the inner layer to the outer layer of the plasma membrane; (b) typical chromatin condensation; and (c) ultrastructural modifications related to apoptotic cell death. We also showed that the expression of p53 was not changed in sulforaphane-treated cells. In contrast, whereas bcl-2 was not detected, we observed increased expression of the proapoptotic protein bax, the release of cytochrome c from the mitochondria to the cytosol, and the proteolytic cleavage of poly(ADP-ribose) polymerase. In conclusion, our results strongly suggest that in addition to the activation of detoxifying enzymes, induction of apoptosis is also involved in the sulforaphane-associated chemoprevention of cancer.

AGO1, QDE-2, and RDE-1 are related proteins required for post-transcriptional gene silencing in plants, quelling in fungi, and RNA interference in animals

Abstract: Introduction of transgene DNA may lead to specific degradation of RNAs that are homologous to the transgene transcribed sequence through phenomena named post-transcriptional gene silencing (PTGS) in plants, quelling in fungi, and RNA interference (RNAi) in animals. It was shown previously that PTGS, quelling, and RNAi require a set of related proteins (SGS2, QDE-1, and EGO-1, respectively). Here we report the isolation of Arabidopsis mutants impaired in PTGS which are affected at the Argonaute1 (AGO1) locus. AGO1 is similar to QDE-2 required for quelling and RDE-1 required for RNAi. Sequencing of ago1 mutants revealed one amino acid essential for PTGS that is also present in QDE-2 and RDE-1 in a highly conserved motif. Taken together, these results confirm the hypothesis that these processes derive from a common ancestral mechanism that controls expression of invading nucleic acid molecules at the post-transcriptional level. As opposed to rde-1 and qde-2 mutants, which are viable, ago1 mutants display several developmental abnormalities, including sterility. These results raise the possibility that PTGS, or at least some of its elements, could participate in the regulation of gene expression during development in plants.

Four Genes of Medicago truncatula Controlling Components of a Nod Factor Transduction Pathway

Abstract: Rhizobium nodulation (Nod) factors are lipo-chitooligosaccharides that act as symbiotic signals, eliciting several key developmental responses in the roots of legume hosts. Using nodulation-defective mutants of Medicago truncatula, we have started to dissect the genetic control of Nod factor transduction. Mutants in four genes (DMI1, DMI2, DMI3, and NSP) were pleiotropically affected in Nod factor responses, indicating that these genes are required for a Nod factor–activated signal transduction pathway that leads to symbiotic responses such as root hair deformations, expressions of nodulin genes, and cortical cell divisions. Mutant analysis also provides evidence that Nod factors have a dual effect on the growth of root hair: inhibition of endogenous (plant) tip growth, and elicitation of a novel tip growth dependent on (bacterial) Nod factors. dmi1, dmi2, and dmi3 mutants are also unable to establish a symbiotic association with endomycorrhizal fungi, indicating that there are at least three common steps to nodulation and endomycorrhization in M. truncatula and providing further evidence for a common signaling pathway between nodulation and mycorrhization.

PROCUSTE1 Encodes a Cellulose Synthase Required for Normal Cell Elongation Specifically in Roots and Dark-Grown Hypocotyls of Arabidopsis

Abstract: Mutants at the PROCUSTE1 (PRC1) locus show decreased cell elongation, specifically in roots and dark-grown hypocotyls. Cell elongation defects are correlated with a cellulose deficiency and the presence of gapped walls. Map-based cloning of PRC1 reveals that it encodes a member (CesA6) of the cellulose synthase catalytic subunit family, of which at least nine other members exist in Arabidopsis. Mutations in another family member, RSW1 (CesA1), cause similar cell wall defects in all cell types, including those in hypocotyls and roots, suggesting that cellulose synthesis in these organs requires the coordinated expression of at least two distinct cellulose synthase isoforms.

Interactions between forest stands and microclimate: Ecophysiological aspects and consequences for silviculture

Abstract: At a local scale, forest trees and stands have a marked influence on climate; thus it is possible to define microclimates. These effects depend on local climatic characteristics and stand type. All climatic parameters should be considered, but partic ular attention should be paid to temperature, light and water. From a silvicultural point of view knowledge of the interactions exis ting between microclimatic conditions and stands, in conjunction with information now available concerning tree ecophysiology make i t possible to produce viable applications which are useful for silviculture during stand formation, and for applying silvicultura l treat- ments. Whitout a doubt, taking forest cover interactions into account (climate and ecophysiological potential of species) is th e basis of sustainable management in forests. Based on the current knowledge about young stands, it is possible to use existing vegetat ion cover, in a more or less modified form, to improve temperature conditions especially in relation to spring frost damage, and to improve water conditions for both plant uptake and plant growth for a variable period depending on the species, but for a minim um of 4 to 5 years. For clearing and thinning, the effects of microclimatic changes created by this type of forestry management, a nd con- sequently the response of trees in terms of photosynthesis and growth, are now well defined. forest stands / microclimate / ecophysiology / silviculture

Biochemical quality of crop residues and carbon and nitrogen mineralization kinetics under nonlimiting nitrogen conditions

Abstract: Statistical relationships were established between the fate of C and N from 47 types of crop residues and their biochemical characteristics during a soil incubation at 15°C. The incubations were carried out under nonlimiting N in order to differentiate the effects of biochemical characteristics of residues from those of soil N availability. Depending on the residue, the apparent mineralization of residue C after 168 d varied from 330 to 670 g kg -1 of added C. Mineralization kinetics were described using a two-compartment decomposition model that decomposes according to first-order kinetics. Amounts of C mineralized after 7 d and the decomposition rate coefficient of the labile fraction were related mainly to the soluble C forms of the residue. No statistical relationship was established between the N concentration of residues and their decomposition in the soil. The incorporation of crop residues into soil led to various soil mineral N dynamics. Two residues caused net N mineralization from the time of their incorporation, whereas all the others induced net N immobilization (1-33 g N kg -1 of added C). After 168 d, only residues with a C/N ratio <24 induced a surplus of mineral N compared with the control soil. The mineral N dynamics were related mainly to the organic N concentration of the residues and to their C/N ratio. At the start of incubation, these dynamics were also influenced by the presence of polyphenols in the plant tissues. Finally, this study showed the need to include the biochemical quality of crop residues in any C and N transformation models that describe decomposition. In contrast, the N concentration or C/N ratio of the residues are sufficient to predict the net effects of crop residues on soil mineral N dynamics.

The bioavailability of dietary calcium.

Abstract: This update focuses on the bioavailability of dietary calcium for humans. Fundamentals of calcium metabolism, intestinal absorption, urinary excretion and balance are recalled. Dietary factors, especially lactose and other milk components, influencing calcium bioavailability at intestinal and renal levels are reviewed. A critical examination of all the methods used for evaluating calcium bioavailability is made. This includes in vitro assays, classical and isotopic balances, urinary excretion, isotope labeling in the urine, plasma and bones, long term evaluation of bone mineralization and the use of biological bone markers. Importance and advantages of animal models are discussed. The state of the art in the comparative bioavailability of calcium in foods is detailed including a comparison of sources of calcium (dairy products and calcium salts) in human studies and in some animal studies, casein phosphopeptides, proteins, lactose and lactase and their relation with calcium bioavailability (in humans and ...