Microbiota in Wheat Roots,Rhizosphere and Soil in Crops Grown in Organic and Other Production Systems

Culturable microbial communities and diseases were compared in organic, integrated and conventional systems of winter wheat production and monoculture. Particular emphasis was placed on the density and diversity of cereal pathogens and their potential antagonists, and on the association of the active microbial populations with the health and productivity of wheat. In roots, rhizoplane and rhizosphere, fungi tended to be most abundant in the integrated system or monoculture, and bacteria in the organic system. The dominant fungal groups (with individual frequency >5%) included root pathogens (Fusarium, Gibberella, Haematonectria and Ilyonectria) and known pathogen antagonists (Acremonium strictum, Clonostachys, Chaetomium, Gliocladium and Trichoderma spp.). The 50 subdominant species (with individual frequency 1–5%) included the pathogens Alternaria, Cladosporium (leaf spot), Gaeumannomyces graminis (take‐all), Glomerella graminicola (anthracnose), Oculimacula yallundae (eyespot), Phoma spp. (leaf spots), and Pythium and Rhizoctonia (root rot). The 40 subrecedent species (with individual frequency <1%) included minor pathogens (Botrytis, Coniothyrium, Leptosphaeria). Antagonists in roots, rhizoplane and rhizosphere were most frequent in the organic system and least frequent in monoculture, suggesting that these systems had the most and least disease‐suppressive habitats, respectively. The other two systems were intermediate, with microbial communities suggesting that the conventional system produced a slightly more suppressive environment than the integrated system. The highest grain yield, in the integrated system, was associated with high abundance of fungi, including fungal pathogens, lowest abundance of Arthrobacter, Pseudomonas and Streptomyces in roots, rhizoplane and soil, and relatively high stem‐base and leaf disease severity. The lowest grain yields, in the organic system and monoculture, were associated with less abundant fungi and more abundant Pseudomonas. There is no clear indication that yields were affected by diseases.     

The anti-genomic (negative) strand of Hepatitis C Virus is not targetable by shRNA

Hepatitis C Virus (HCV) and other plus-strand RNA viruses typically require the generation of a small number of negative genomes (20–100× lower than the positive genomes) for replication, making the less-abundant antigenome an attractive target for RNA interference(RNAi)-based therapy. Because of the complementarity of duplex short hairpin RNA/small interfering RNA (shRNA/siRNAs) with both genomic and anti-genomic viral RNA strands, and the potential of both shRNA strands to become part of the targeting complexes, preclinical RNAi studies cannot distinguish which viral strand is actually targeted in infected cells. Here, we addressed the question whether the negative HCV genome was bioaccessible to RNAi. We first screened for the most active shRNA molecules against the most conserved regions in the HCV genome, which were then used to generate asymmetric anti-HCV shRNAs that produce biologically active RNAi specifically directed against the genomic or antigenomic HCV sequences. Using this simple but powerful and effective method to screen for shRNA strand selectivity, we demonstrate that the antigenomic strand of HCV is not a viable RNAi target during HCV replication. These findings provide new insights into HCV biology and have important implications for the design of more effective and safer antiviral RNAi strategies seeking to target HCV and other viruses with similar replicative strategies.     

Beta-amyloid peptides enhance the proliferative response of activated CD4CD28 lymphocytes from Alzheimer disease patients and from healthy elderly

Alzheimer's disease (AD) is the most frequent form of dementia among elderly. Despite the vast amount of literature on non-specific immune mechanisms in AD there is still little information about the potential antigen-specific immune response in this pathology. It is known that early stages of AD include β-amyloid (Aβ)- reactive antibodies production and inflammatory response. Despite some evidence gathered proving cellular immune response background in AD pathology, the specific reactions of CD4(+) and CD8(+) cells remain unknown as the previous investigations yielded conflicting results. Here we investigated the CD4(+)CD28(+) population of human peripheral blood T cells and showed that soluble β-amyloids alone were unable to stimulate these cells to proliferate significantly, resulting only in minor, probably antigen-specific, proliferative response. On the other hand, the exposure of in vitro pre-stimulated lymphocytes to soluble Aβ peptides significantly enhanced the proliferative response of these cells which had also lead to increased levels of TNF, IL-10 and IL-6. We also proved that Aβ peptide-enhanced proliferative response of CD4(+)CD28(+) cells is autonomous and independent from disease status while being associated with the initial, ex vivo activation status of the CD4(+) cells. In conclusion, we suggest that the effect of Aβ peptides on the immune system of AD patients does not depend on the specific reactivity to Aβ epitope(s), but is rather a consequence of an unspecific modulation of the cell cycle dynamics and cytokine production by T cells, occurring simultaneously in a huge proportion of Aβ peptide-exposed T lymphocytes and affecting the immune system performance.     

The use of supramolecular structures as protein ligands

Congo red dye as well as other eagerly self-assembling organic molecules which form rod-like or ribbon-like supramolecular structures in water solutions, appears to represent a new class of protein ligands with possible wide-ranging medical applications. Such molecules associate with proteins as integral clusters and preferentially penetrate into areas of low molecular stability. Abnormal, partly unfolded proteins are the main binding target for such ligands, while well packed molecules are generally inaccessible. Of particular interest is the observation that local susceptibility for binding supramolecular ligands may be promoted in some proteins as a consequence of function-derived structural changes, and that such complexation may alter the activity profile of target proteins. Examples are presented in this paper.     

Resveratrol,a naturally occurring diphenolic compound,affects lipogenesis,lipolysis and the antilipolytic action of insulin in isolated rat adipocytes

Resveratrol is a naturally occurring diphenolic compound exerting numerous beneficial effects in the organism. The present study demonstrated its short-term, direct influence on lipogenesis, lipolysis and the antilipolytic action of insulin in freshly isolated rat adipocytes. In fat cells incubated for 90 min with 125 and 250 μM resveratrol (but not with 62.5 μM resveratrol), basal and insulin-induced lipogenesis from glucose was significantly reduced. The antilipogenic effect was accompanied by a significant diminution of CO₂ release and enhanced production of lactate. The inhibition of glucose conversion to lipids found in the presence of resveratrol was not attenuated by activator of protein kinase C. However, acetate conversion to lipids appeared to be insensitive to resveratrol.In adipocytes incubated for 90 min with epinephrine, 10 and 100 μM resveratrol significantly enhanced lipolysis, especially at lower concentrations of the hormone. However, the lipolytic response to dibutyryl-cAMP, a direct activator of protein kinase A, was unchanged. Further studies demonstrated that, in cells stimulated with epinephrine, 1, 10 and 100 μM resveratrol significantly enhanced glycerol release despite the presence of insulin or H-89, an inhibitor of protein kinase A. The influence of resveratrol on epinephrine-induced lipolysis and on the antilipolytic action of insulin was not abated by the blocking of estrogen receptor and was accompanied by a significant (with the exception of 1 μM resveratrol in experiment with insulin) increase in cAMP in adipocytes. It was also revealed that resveratrol did not change the proportion between glycerol and fatty acids released from adipocytes exposed to epinephrine.Results of the present study revealed that resveratrol reduced glucose conversion to lipids in adipocytes, probably due to disturbed mitochondrial metabolism of the sugar. Moreover, resveratrol increased epinephrine-induced lipolysis. This effect was found also in the presence of insulin and resulted from the synergistic action of resveratrol and epinephrine. The obtained results provided evidence that resveratrol affects lipogenesis and lipolysis in adipocytes contributing to reduced lipid accumulation in these cells.     

Radioimmunologic proinsulin determination in the serum using an "insulin-catabolizing protease" from the rat liver]

Proinsulin determinations in human serum are of particular interest, because proinsulin represents only a fraction of the biological activity of insulin. Proinsulin like components are determined by means of an "Insulin Degrading Protease" (ISP) which degrades insulin into non-radioimmunoassayable fission products. The radioimmunoassay before and after incubation with this enzyme provides values for the total-insulin and the proinsulin. The preparation of the ISP is done by homogenization and ultracentrifugation of fresh liver tissue followed by dialysation and dryfreezing. After further concentration by adsorption to a calcium-phosphate-gel the ISP degrades pure porcine insulin within 20' down to a rest of 9%, but only 7% of pure porcine proinsulin is altered. The proinsulin values provided this way are reproducible and exact enough for the clinical use. They correspond largely to those methods using chromatographic columns. In 13 persons the proinsulin fraction of the total insulin after stimulation with glucose and tolbutamid has been registered. The proinsulin shows in the oral glucose tolerance test compared to insulin a lower and delayed increase. After tolbutamid only minor changes of proinsulin values have been seen. As long as it is difficult to prepare a proinsulin specific antibody for a direct proinsulin radioimmunoassay, the ISP-method is even qualified for extensive proinsulin determinations in human serum.     

Effects of hydrogen, oxygen, and ammonia low-pressure glow plasma on granular starches

Cassava, corn, high amylose corn, potato, rice Indica, rice Japonica, sweet potato, waxy corn, and wheat starches were exposed to low-pressure ammonia, hydrogen, and oxygen plasma. In every case, depolymerization of the starch polysaccharides was noted. The extent of the depolymerization depended on the nature of the starch as well as the type of plasma applied. Among three fractions of polysaccharides distinguished by their molecular weight average, the fraction of the highest molecular weight suffered the most efficient depolymerization. The relative depolymerization for the middle- and low-molecular fractions of polysaccharides was found to be starch and plasma specific. The chemical character of the plasma had very little influence on the starch polysaccharides. Only subtle oxidation effects were observed in oxygen plasma. Low-pressure glow plasma treatment appeared to be a convenient tool for a waste-less dextrinization of starch. Manipulation of the plasma variety and the time of exposure resulted in a wide spectrum of dextrins of various molecular weights and paste-forming properties.