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951.
Bertazzon N Raiola A Castiglioni C Gardiman M Angelini E Borgo M Ferrari S 《Plant cell reports》2012,31(1):133-143
Grapevine is an economically important crop, and the recent completion of its genome makes it possible to study the function
of specific genes through reverse genetics. However, the analysis of gene function by RNA interference (RNAi) in grapevine
is difficult, because the generation of stable transgenic plants has low efficiency and is time consuming. Recently, transient
expression of genes in grapevine leaves has been obtained by Agrobacterium tumefaciens infiltration (agroinfiltration). We therefore tested the possibility to silence grapevine genes by agroinfiltration of RNAi
constructs. A construct to express a double strand RNA (dsRNA) corresponding to the defense-related gene VvPGIP1, encoding a polygalacturonase-inhibiting protein (PGIP), was obtained and transiently expressed by agroinfiltration in leaves
of grapevine plants grown in vitro. Expression of VvPGIP1 and accumulation of PGIP activity were strongly induced by infiltration with control bacteria, but not with bacteria carrying
the dsRNA construct, indicating that the gene was efficiently silenced. In contrast, expression of another defense-related
gene, VST1, encoding a stilbene synthase, was unaffected by the dsRNA construct. We have therefore demonstrated the possibility of transient
down-regulation of grapevine genes by agroinfiltration of constructs for the expression of dsRNA. This system can be employed
to evaluate the effectiveness of constructs that can be subsequently used to generate stable RNAi transgenic plants. 相似文献
952.
The flavone luteolin was extracted, isolated and purified from leaves of Cichorium endivia L. subsp. divaricatum and identified based on 1H and 13C NMR spectral analyses. Luteolin (0.0015?%) was applied to two cultivars of maize (Giza 2 and single cross 10) grains, germinated in the absence and presence of NaCl, in order to assess the bioactivity of luteolin and its role in alleviating the harmful effect of salt stress on maize. The results showed that luteolin enhanced germination and seedling growth of both maize cultivars in normal conditions. Furthermore, luteolin successfully alleviated the harmful effect of salinity on germination and seedling growth of maize. The potential role of luteolin was performed mainly by stimulating ??-amylase activity leading to enhancing starch mobilization and increasing the accumulation of soluble sugars, and partially by enhancing the antioxidative defense. As future prospective, a great number of phenolic compounds need to be screened in order to make use of them in improving agriculture under stressful conditions. 相似文献
953.
Nadia Danilova Elena Bibikova Todd M. Covey David Nathanson Elizabeth Dimitrova Yoan Konto Anne Lindgren Bertil Glader Caius G. Radu Kathleen M. Sakamoto Shuo Lin 《Disease models & mechanisms》2014,7(7):895-905
Ribosomal biogenesis involves the processing of pre-ribosomal RNA. A deficiency of some ribosomal proteins (RPs) impairs processing and causes Diamond Blackfan anemia (DBA), which is associated with anemia, congenital malformations and cancer. p53 mediates many features of DBA, but the mechanism of p53 activation remains unclear. Another hallmark of DBA is the upregulation of adenosine deaminase (ADA), indicating changes in nucleotide metabolism. In RP-deficient zebrafish, we found activation of both nucleotide catabolism and biosynthesis, which is consistent with the need to break and replace the faulty ribosomal RNA. We also found upregulation of deoxynucleotide triphosphate (dNTP) synthesis – a typical response to replication stress and DNA damage. Both RP-deficient zebrafish and human hematopoietic cells showed activation of the ATR/ATM-CHK1/CHK2/p53 pathway. Other features of RP deficiency included an imbalanced dNTP pool, ATP depletion and AMPK activation. Replication stress and DNA damage in cultured cells in non-DBA models can be decreased by exogenous nucleosides. Therefore, we treated RP-deficient zebrafish embryos with exogenous nucleosides and observed decreased activation of p53 and AMPK, reduced apoptosis, and rescue of hematopoiesis. Our data suggest that the DNA damage response contributes to p53 activation in cellular and zebrafish models of DBA. Furthermore, the rescue of RP-deficient zebrafish with exogenous nucleosides suggests that nucleoside supplements could be beneficial in the treatment of DBA.KEY WORDS: Ribosomal protein deficiency, Rps19, Rpl11, p53, ATR, RNR, Chk1, ATP, AMPK, Exogenous nucleosides 相似文献
954.
Natha J. Booth Judith B. Beekman Ronald L. Thune 《Applied and environmental microbiology》2009,75(21):6712-6720
Genomic analysis indicated that Edwardsiella ictaluri encodes a putative urease pathogenicity island containing the products of nine open reading frames, including urea and ammonium transporters. In vitro studies with wild-type E. ictaluri and a ureG::kan urease mutant strain indicated that E. ictaluri is significantly tolerant of acid conditions (pH 3.0) but that urease activity is not required for acid tolerance. Growth studies demonstrated that E. ictaluri is unable to grow at pH 5 in the absence of urea but is able to elevate the environmental pH from pH 5 to pH 7 and grow when exogenous urea is available. Substantial production of ammonia was observed for wild-type E. ictaluri in vitro in the presence of urea at low pH, and optimal activity occurred at pH 2 to 3. No ammonia production was detected for the urease mutant. Proteomic analysis with two-dimensional gel electrophoresis indicated that urease proteins are expressed at both pH 5 and pH 7, although urease activity is detectable only at pH 5. Urease was not required for initial invasion of catfish but was required for subsequent proliferation and virulence. Urease was not required for initial uptake or survival in head kidney-derived macrophages but was required for intracellular replication. Intracellular replication of wild-type E. ictaluri was significantly enhanced when urea was present, indicating that urease plays an important role in intracellular survival and replication, possibly through neutralization of the acidic environment of the phagosome.Identification of virulence factors is vitally important to an understanding of the pathogenesis of Edwardsiella ictaluri and to the development of methods for controlling the spread of disease. Although the pathogenesis of E. ictaluri was reviewed in 1993 (28, 31), recent reports demonstrated that E. ictaluri is a facultative intracellular pathogen (3) and that a type III secretion system is required for intracellular survival and replication within channel catfish head kidney-derived macrophages (HKDM) (30). Using signature-tagged mutagenesis (STM) in an immersion challenge model for E. ictaluri, Thune et al. (30) identified 50 transconjugants carrying transposon insertions in genes required for survival and replication in the channel catfish host. Two of those mutants had insertions in genes encoding homologs of UreG and UreF, proteins that are essential for the production of an active urease enzyme in other bacteria (6, 10, 14, 26). UreG is a GTP-binding accessory protein that functions in energy-dependent assembly of the urease holoenzyme (19), while UreF is a urease accessory protein that functions in the generation or delivery of carbon dioxide to the urease metallocenter assembly site (19). Both the ureG and ureF mutant strains were further characterized in a competitive challenge with the wild-type (WT) parental strain and were confirmed to be significantly attenuated (30). The identification of two mutants with insertions in urease-associated genes suggests an important role for urease activity in E. ictaluri pathogenesis, despite the fact that E. ictaluri is urease negative in standard biochemical tests. Consequently, the objectives of this study are to characterize the E. ictaluri urease pathogenicity island (PAI), to evaluate conditions for E. ictaluri urease activity, and to establish a possible role for urease in E. ictaluri pathogenesis. 相似文献
955.
Annett Mikolasch Veronika Hahn Katrin Manda Judith Pump Nicole Illas Dirk Gördes Michael Lalk Manuela Gesell Salazar Elke Hammer Wolf-Dieter Jülich Stephan Rawer Kerstin Thurow Ulrike Lindequist Frieder Schauer 《Amino acids》2010,39(3):671-683
In order to design potential biomaterials, we investigated the laccase-catalyzed cross-linking between l-lysine or lysine-containing peptides and dihydroxylated aromatics. l-Lysine is one of the major components of naturally occurring mussel adhesive proteins (MAPs). Dihydroxylated aromatics are structurally related to 3,4-dihydroxyphenyl-l-alanine, another main component of MAPs. Mass spectrometry and nuclear magnetic resonance analyses show that the ε-amino group of l-lysine is able to cross-link dihydroxylated aromatics. Additional oligomer and polymer cross-linked products were obtained from di- and oligopeptides containing l-lysine. Potential applications in medicine or industry for biomaterials synthesised via the three component system consisting of the oligopeptide [Tyr-Lys]10, dihydroxylated aromatics and laccase are discussed. 相似文献
956.
Judith A. C. Rietjens Paul J. van der Maas Bregje D. Onwuteaka-Philipsen Johannes J. M. van Delden Agnes van der Heide 《Journal of bioethical inquiry》2009,6(3):271-283
Two decades of research on euthanasia in the Netherlands have resulted into clear insights in the frequency and characteristics
of euthanasia and other medical end-of-life decisions in the Netherlands. These empirical studies have contributed to the
quality of the public debate, and to the regulating and public control of euthanasia and physician-assisted suicide. No slippery
slope seems to have occurred. Physicians seem to adhere to the criteria for due care in the large majority of cases. Further,
it has been shown that the majority of physicians think that the euthanasia Act has improved their legal certainty and contributes
to the carefulness of life-terminating acts. In 2005, eighty percent of the euthanasia cases were reported to the review committees.
Thus, the transparency envisaged by the Act still does not extend to all cases. Unreported cases almost all involve the use
of opioids, and are not considered to be euthanasia by physicians. More education and debate is needed to disentangle in these
situations which acts should be regarded as euthanasia and which should not. Medical end-of-life decision-making is a crucial
part of end-of-life care. It should therefore be given continuous attention in health care policy and medical training. Systematic
periodic research is crucial for enhancing our understanding of end-of-life care in modern medicine, in which the pursuit
of a good quality of dying is nowadays widely recognized as an important goal, in addition to the traditional goals such as
curing diseases and prolonging life. 相似文献
957.
Hooper C Day R Slocombe R Handlinger J Benkendorff K 《Fish & shellfish immunology》2007,22(4):363-379
Increasing mariculture of abalone focuses attention on their immune and stress responses. For abalone, as well as many invertebrates, the function and relationship of these systems and how in vitro tests relate to them are not fully understood. This review focuses on research into the immune system and stress response conducted on abalone and on aspects that can be monitored in vitro. To fill the considerable knowledge gaps, we discuss work on other invertebrate taxa, concentrating on those closest to abalone, and making explicit the phylogenetic relations involved. The stress response appears to be very similar to that in vertebrates, but interpreting most immune responses remains problematic. Phylogeny must be considered: immune function tests derived from research into vertebrates or distantly related invertebrates should not be used in abalone until they have been validated in abalone by studies of susceptibility to pathogens. We suggest phagocytic activity of haemocytes and their efficiency in clearing bacteria are reliable parameters to measure, because they have been directly related to immune competency and are consistently depressed by stress. Carefully designed assays of antimicrobial activity may also be useful. Important aims of future research will be to investigate the relationship between growth, stress and robust immunity, and to develop tests that can be run on production animals, which accurately depict immune status. 相似文献
958.
Singleton K Parvaze N Dama KR Chen KS Jennings P Purtic B Sjaastad MD Gilpin C Davis MM Wülfing C 《Journal of immunology (Baltimore, Md. : 1950)》2006,177(7):4402-4413
T cell activation is driven by the TCR and complemented by costimulation. We have studied the dynamics of ligand-engagement of the costimulatory receptor CD2 in T cell/APC couples. Thousands of ligand-engaged CD2 molecules were included in a large T cell invagination at the center of the cellular interface within 1 min of cell couple formation. The structure and regulation of this invagination shared numerous features with phagocytosis and macropinocytosis. Three observations further characterize the invagination and the inclusion of CD2: 1) numerous ligand-engaged receptors were enriched in and internalized through the T cell invagination, none as prominently as CD2; 2) dissolution of the T cell invagination and CD2 engagement were required for effective proximal T cell signaling; and 3) the T cell invagination was uniquely sensitive to the affinity of the TCR for peptide-MHC. Based on this characterization, we speculate that the T cell invagination, aided by CD2 enrichment, internalizes parts of the TCR signaling machinery to reset T cell signaling upon agonist-mediated, stable APC contact. 相似文献
959.
Walton KA Gugiu BG Thomas M Basseri RJ Eliav DR Salomon RG Berliner JA 《Journal of lipid research》2006,47(9):1967-1974
Previous studies from our laboratory and others presented evidence that oxidized 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphatidylcholine (OxPAPC) and oxidized 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphatidylethanolamine can inhibit lipopolysaccharide (LPS)-mediated induction of interleukin-8 (IL-8) in endothelial cells. Using synthetic derivatives of phosphatidylethanolamine, we now demonstrate that phospholipid oxidation products containing alpha,beta-unsaturated carboxylic acids are the most active inhibitors we examined. 5-Keto-6-octendioic acid ester of 2-phosphatidylcholine (KOdiA-PC) was 500-fold more inhibitory than OxPAPC, being active in the nanomolar range. Our studies in human aortic endothelial cells identify one important mechanism of the inhibitory response as involving the activation of neutral sphingomyelinase. There is evidence that Toll-like receptor-4 and other members of the LPS receptor complex must be colocalized to the caveolar/lipid raft region of the cell, where sphingomyelin is enriched, for effective LPS signaling. Previous work from our laboratory suggested that OxPAPC could disrupt this caveolar fraction. These studies present evidence that OxPAPC activates sphingomyelinase, increasing the levels of 16:0, 22:0, and 24:0 ceramide and that the neutral sphingomyelinase inhibitor GW4869 reduces the inhibitory effect of OxPAPC and KOdiA-PC. We also show that cell-permeant C6 ceramide, like OxPAPC, causes the inhibition of LPS-induced IL-8 synthesis and alters caveolin distribution similar to OxPAPC. Together, these data identify a new pathway by which oxidized phospholipids inhibit LPS action involving the activation of neutral sphingomyelinase, resulting in a change in caveolin distribution. Furthermore, we identify specific oxidized phospholipids responsible for this inhibition. 相似文献
960.
Elinor M. Lichtenberg Sarah K. Richman Rebecca E. Irwin Judith L. Bronstein 《Ecological Entomology》2020,45(4):904-909
1. Competition alters animal foraging, including promoting the use of alternative resources. It may also impact how animals feed when they are able to handle the same food with more than one tactic. Competition likely impacts both consumers and their resources through its effects on food handling, but this topic has received little attention. 2. Bees often use two tactics for extracting nectar from flowers: they can visit at the flower opening, or rob nectar from holes at the base of flowers. Exploitative competition for nectar is thought to promote nectar robbing. If so, higher competition among floral visitors should reduce constancy to a single foraging tactic as foragers will seek food using all possible tactics. To test this prediction, field observations and two experiments involving bumble bees visiting three montane Colorado plant species (Mertensia ciliata, Linaria vulgaris, Corydalis caseana) were used under various levels of inter- and intra-specific competition for nectar. 3. In general, individual bumble bees remained constant to a single foraging tactic, independent of competition levels. However, bees that visited M. ciliata in field observations decreased their constancy and increased nectar robbing rates as visitation rates by co-visitors increased. 4. While tactic constancy was high overall regardless of competition intensity, this study highlights some intriguing instances in which competition and tactic constancy may be linked. Further studies investigating the cognitive underpinnings of tactic constancy should provide insight on the ways in which animals use alternative foraging tactics to exploit resources. 相似文献