Oxidative stress induced by long-term plum pox virus infection in peach (Prunus persica)

In this study, the effect of long-term plum pox virus (PPV) infection on the response of certain antioxidant enzymes at the subcellular level was studied in peach plants ( Prunus persica (L.) Batch) (cv. GF305), which are characterized by great susceptibility to the virus. In infected plants, a decrease in the efficiency of excitation energy capture by PSII ( F _v'/ F _m') was observed, which was accompanied by a decrease in non-photochemical quenching (NPQ). p -Hydroxy-mercury benzoic acid (pHMB)-insensitive ascorbate peroxidase (APX) activity (class III peroxidase) was detected in both chloroplast and soluble fractions. In soluble fractions from inoculated peaches, a significant increase in pHMB-sensitive APX activity and a significant decrease in superoxide dismutase (SOD) activity were observed. These changes were correlated with the observations in isolated chloroplasts, where an increase in both pHMB-sensitive and pHMB-insensitive APX activities was observed, whereas significant decreases in SOD, monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR) activities were produced. According to these results, as a consequence of PPV infection, an oxidative stress, indicated by an increase in lipid peroxidation and protein oxidation, was produced in peach leaves, which was monitored by the diaminobenzidine (DAB) peroxidase-coupled H₂O₂ probe. PPV infection produced an alteration in chloroplast ultrastructure, giving rise to dilated thylakoid membranes. PPV-infected peach leaves showed a decreased amount of starch in chloroplasts from palisade parenchyma, as well as an increase in the number and size of plastoglobuli, in relation to control plants. The results suggest that long-term PPV infection produces an oxidative stress, and that an antioxidative metabolism imbalance may be related to the progress of PPV infection and symptoms in peach plants.     

Natriuretic Peptide Clearance Receptor Alleles and Susceptibility to Abdominal Adiposity

Objective: To test the association of the C(?55)A polymorphism of the natriuretic peptide clearance receptor (NPRC) with blood pressure (BP), overweight/obesity, and body fat distribution in a large male adult population. Research Methods and Procedures: The study population was from a cross‐sectional and follow‐up study of 787 untreated male participants in the 1994 to 1995 follow‐up examination of the Olivetti Heart Study in Naples (356 of whom were examined previously in 1975). BP and anthropometric measures were taken, and biochemical assays were performed. The NPRC gene polymorphism C(?55)A was evaluated by polymerase chain reaction and HgaI digestion. Results: In the whole study population, there was no difference in BP, BMI, and biochemical tests among genotypes. Considering an A(?55) recessive model of inheritance, the AA subjects had lower BMI and waist circumference and lower prevalence of overweight, obesity, and abdominal adiposity as compared with the CC+CA subjects. On reviewing the characteristics of the subgroup previously examined in 1975, the AA subjects had already lower BMI, and their 20‐year rate of overweight and obesity was lower than the CC+CA subjects; no difference was observed in the rate of hypertension. Discussion: Male subjects carrying the A(?55)A NPRC genotype had a significantly lower prevalence of overweight, obesity, and abdominal adiposity. They also had a lower 20‐year rate of overweight compared with CC+CA individuals. These findings from a large unselected and untreated male population are in keeping with the recent evidence of a powerful lipolytic and lipomobilizing activity of natriuretic peptides.     

Effect of N-acetyl- -cysteine on peroxynitrite and superoxide anion production of lung alveolar macrophages in systemic sclerosis

Lung macrophages may play a relevant role in oxidative processes producing both superoxide anion (O(2)(-)) and NO. In this view, an antioxidant therapy can be useful in the treatment of systemic sclerosis (SSc) patients. N-Acetylcysteine (NAC) is able to expand natural antioxidant defenses by increasing intracellular gluthatione concentration and it has been proposed as an antioxidant therapy in respiratory distress syndromes. The aim of our study was to determine whether lung macrophages obtained from SSc patient bronchoalveolar lavage (BAL) express the inducible form of nitric oxide synthase (iNOS) and whether NAC can reduce the peroxynitrite (ONOO(-)) and O(2)(-) production of these cells. Alveolar macrophages were isolated from BAL of 32 patients and used for the immunocytochemical determination of iNOS, and the production of ONOO(-) and O(2)(-) was measured by fluorimetric or spectrophotometric methods, respectively. Lung macrophages obtained from SSc patients expressed a higher level of iNOS compared to healthy subject cells. NAC preincubation (5 x 10(-5)M, 24h) significantly reduced (-21%) the ONOO(-) production in formyl Met-Leu-Phe (fMLP)-activated cells and slightly reduced it under resting conditions, whereas NAC preincubation was unable to modify the release of O(2)(-) both in basal condition and in fMLP-stimulated cells. We conclude that since SSc lung macrophages express high levels of iNOS and produce a significant quantity of ONOO(-), NAC administration reduces ONOO(-) production and can be an useful treatment to alleviate SSc symptoms.     

The impact of H2-DM on humoral immune responses.

H2-DM (DM, previously H2-M) facilitates the exchange of peptides bound to MHC class II molecules. In this study, we have used H2-DM-deficient (DM(-/-)) mice to analyze the influence of DM in the priming of B cell responses in vivo and for Ag presentation by B cells in vitro. After immunization, IgG Abs could be raised to a T-dependent Ag, 4-hydroxy-5-nitrophenylacetyl-OVA, in DM(-/-) mice, but closer analysis revealed the IgG response to be slower, diminished in titer, and composed of low-affinity Abs. The Ab response correlated with a vast reduction in the number of germinal centers in the spleen. The presentation of multiple epitopes by H2-A(b) from distinct Ags was found to be almost exclusively DM-dependent whether B cells internalized Ags via fluid phase uptake or using membrane Ig receptors. The poor B cell response in vivo could be largely, but not completely restored by expression of a H2-Ea(d) transgene, despite the fact that Ag presentation by H2-E(d/b) molecules was found to be highly DM dependent. Hence, while substantial Ab responses can be raised in the absence of DM, this molecule is a crucial factor both for Ag processing and for the normal maturation of T-dependent humoral immune responses in vivo.     

Plant resources of the Tehuacán-Cuicatlán valley, Mexico

Information on richness of plant resources, and their forms of use and management in the biosphere reserve Tehuacan-Cuicatlan, Mexico is analyzed. This 10 000 km² region hosts nearly 2700 vascular plant species, and it is acknowledged as one of the arid areas with the highest floristic diversity in North America. The seven indigenous ethnic groups that live in this region have cultural roots that date back almost 10 000 years. Based upon ethnobotanical and floristic studies, as well as bibliographical sources, a total of 808 useful plant species were identified, most of them (90%) being native, and 44 species being endemic to the region. A total of 681 species are wild plants, 109 are weeds and ruderal plants, and 86 are domesticated crops. However, it was noted that considerable overlap exists between the species of these 3 categories. For example, while wild and ruderal plants (706 species) are foraged by both humans and domestic animals, 59 species of this group are also managed in situ. On the other hand, 168 wild, ruderal and domesticated species are cultivated. The Tehuacán-Cuicatlan Valley is one of the richest regions of Mexico in plant resources. Local knowledge on use and management of plants is a valuable source of information for designing conservation and social development strategies for the biosphere reserve.     

Increase of rotifer diversity after sewage diversion in the hypertrophic lagoon,Albufera of Valencia,Spain

The Albufera of Valencia is a large oligohaline hypertrophic lagoon, regulated by sluice gates according to the needs of the surrounding rice field cultivation. It is in a turbid state with permanent cyanobacterial blooms. A slight improvement was detected after diversion in the 1990s of part of the sewage flowing into it. After sewage diversion, we found that: (1) Chlorophyll concentration and rotifer densities decreased; (2) Rotifer proportions declined, due mainly to a relative increase in cladocerans; (3) Rotifer diversity increased. The two dominants of the 1980s, Polyarthra spp. in the colder period and Brachionus angularis in the warmer one, reverted after sewage diversion to a more diverse assemblage reminiscent of the 1970s, with a higher number of dominant species. In the summer of 1998, both Brachionus calyciflorus and its predator Asplanchna brightwelli, dominant in 1973, became abundant again. In 1998, an increase in the number of dominant species was also observed during water renewal periods, some of these species were new or seldomly found before in the lagoon (Proalides tentaculatus-digitus, Trichocerca pusilla at the end of rice culture, Brachionus variabilis at the end of winter flooding). Another change that indicates an improvement of water conditions is a more distinct and longer clear water phase, which occurs in the water renewal period at the end of winter and involves a Daphnia magna peak. The increased importance of this phase, promoted the flourishment of Brachionus variabilis, a facultative Daphnia epibiont never found before in the lake.     

Pigment Production by Streptococcus agalactiae in Quasi-Defined Media

A quasi-defined medium that supports the growth of Streptococcus agalactiae as pigmented colonies has been developed. The medium contains starch, a peptic digest of albumin, amino acids, nucleosides, vitamins, and salts. The presence of free cysteine, which could be replaced with other sulphur-containing compounds and to a lesser degree by reducing agents, was required for pigment formation.     

Molecular phylogeny ofCheirolophus (Asteraceae:Cardueae-Centaureinae) based on ITS sequences of nuclear ribosomal DNA

The genusCheirolophus has an interesting western Mediterranean and Macaronesian distribution. Here we investigate the delimitation of the genus and its exclusion from the large genusCentaurea, the systematic position of the related genusPaleocyanus, the delimitation of some species and the phylogeny of the group. We have carried out a phylogenetic analysis of the PCR-generated sequences of the internal transcribed spacers (ITS-1 and ITS-2) of the nuclear ribosomal DNA. The results suggest that the genus, includingPaleocyanus crassifolius is monophyletic; thus, a new combination of this species underCheirolophus is proposed. The Macaronesian group of species is also monophyletic, indicating a single colonization of the archipelago. The poor resolution of microspecies in the Macaronesian group reinforces the hypothesis of a very recent differentiation of the group.     

Maf/ham1-like pyrophosphatases of non-canonical nucleotides are host-specific partners of viral RNA-dependent RNA polymerases

Cassava brown streak disease (CBSD), dubbed the “Ebola of plants”, is a serious threat to food security in Africa caused by two viruses of the family Potyviridae: cassava brown streak virus (CBSV) and Ugandan (U)CBSV. Intriguingly, U/CBSV, along with another member of this family and one secoviridae, are the only known RNA viruses encoding a protein of the Maf/ham1-like family, a group of widespread pyrophosphatase of non-canonical nucleotides (ITPase) expressed by all living organisms. Despite the socio-economic impact of CDSD, the relevance and role of this atypical viral factor has not been yet established. Here, using an infectious cDNA clone and reverse genetics, we demonstrate that UCBSV requires the ITPase activity for infectivity in cassava, but not in the model plant Nicotiana benthamiana. HPLC-MS/MS experiments showed that, quite likely, this host-specific constraint is due to an unexpected high concentration of non-canonical nucleotides in cassava. Finally, protein analyses and experimental evolution of mutant viruses indicated that keeping a fraction of the yielded UCBSV ITPase covalently bound to the viral RNA-dependent RNA polymerase (RdRP) optimizes viral fitness, and this seems to be a feature shared by the other members of the Potyviridae family expressing Maf/ham1-like proteins. All in all, our work (i) reveals that the over-accumulation of non-canonical nucleotides in the host might have a key role in antiviral defense, and (ii) provides the first example of an RdRP-ITPase partnership, reinforcing the idea that RNA viruses are incredibly versatile at adaptation to different host setups.