Inhibition of photophosphorylation and electron transport chain in thylakoids by lasiodiplodin, a natural product from Botryosphaeria rhodina

Four natural products were isolated from the fungus Botryosphaeria rhodina, and their effects on photosynthesis were tested. Only lasiodiplodin (1) inhibited ATP synthesis and electron flow from water to methylviologen; therefore, it acts as a Hill reaction inhibitor in freshly lysed spinach thylakoids. Photosystem I and II and partial reactions as well as ATPase were measured in the presence of 1. Three new different sites of 1 interaction and inhibition were found: one at CF1, the second in the water-splitting enzyme, and the third at the electron-transfer path between P680 and QA; these targets are different from that of the synthetic herbicides present. Electron transport chain inhibition by 1 was corroborated by fluorescence induction kinetics studies.     

Bioactive Compounds and Antioxidant Capacity of Strawberry Jams

Strawberries represent the main source of ellagic acid derivatives in the Brazilian diet. They are also good sources of flavonoids, mainly anthocyanins, and phenolic acids, to which many beneficial effects have been attributed. However, as the fruit is not available all the year, the objective of this work was to determine whether the jams could also represent a good source of bioactive compounds. In the current study, five different commercially available strawberry jams were characterized in relation to flavonoids, total phenolics, free and total ellagic acid contents, and antioxidant capacity. Anthocyanins were detected only in two jams at very low content. Kaempferol glycosides were the main flavonoids present (from 0.38 to 1.05 mg/100 g fresh weight, FW), while quercetin glycosides were present in the range 0.14–1.20 mg/100 g FW. Free and total ellagic acid content ranged from 0.4 to 2.9 mg/100 g FW, and from 17.0 to 29.5 mg/100 g FW, respectively. Total phenolics varied from 58 to 136 mg/100 g FW, and the antioxidant capacity from 0.55 to 0.76 μmol BHT (Butylhydroxytoluene) equivalents/g FW. Overall, results indicated that jams can also represent a good source of antioxidant compounds, although compared to the fruit important losses seem to occur.     

Antioxidant and antimutagenic properties of Hibiscus tiliaceus L. methanolic extract

The genus Hibiscus thrives in a variety of climates and produces a diversity of natural compounds with bioactive properties. We have studied the chemical composition and the in vivo antioxidant properties of Hibiscus tiliaceus L. methanolic flower extract, as well as its mutagenic/antimutagenic effects. Vitamin E and some stigmasterol derivatives that might confer an antioxidant effect to the extract were present. Treatment with this extract protected several Saccharomyces cerevisiae strains defective in antioxidant defenses against H2O2 and t-BOOH cytotoxicities, showing a clear antioxidant activity. The effect is the same for all strains used, independent of the antioxidant defense disrupted, suggesting that protection may be due to molecules that act as versatile and wide spectrum nonenzymatic antioxidants, such as vitamins or phytosterols. The extract was not mutagenic in either Salmonella typhimurium or S. cerevisiae and showed a significant antimutagenic action against oxidative mutagens in S. cerevisiae.     

Kinetics of energy source utilization in Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) embryonic development

The present work evaluates the kinetics of utilization of the main potential energy sources throughout the embryonic developmental stages of Boophilus microplus. The embryonic development of this arthropod is completed in 21 days. Cellularization of the blastoderm occurs on the 6th day and is rapidly followed by germ band extension and segmentation, whose first signs are visible on the 7th day. Cellularization is typically a maternal-driven process, carried out by molecular determinants deposited in the oocyte during oogenesis. On the other hand, segmentation is of zygotic nature, being the consequence of the synthesis of various components by the growing embryo. The enhancement in total B. microplus RNA was observed after cellularization, corroborating the replacement of maternal-driven processes by embryonic zygotic expression. An abrupt increase in oxygen consumption was observed from cellularization until the 8th day of development. The reduction in dry weight at the same period and the susceptibility of oxygen consumption to KCN suggest that the respiration process is activated during early embryonic development. A marked decrease in total lipid content occurred between the 5th and 7th days of development, suggesting this is the main energy source for cellularization. A major reduction in carbohydrate content occurred later, between the 7th and 9th days, and it could be assigned to the morphological segmentation of the embryo. Although the total amount of proteins remains unchanged from oviposition to hatching, a 15% reduction in vitellin (VT) content was observed before cellularization, up to the 4th day after egglaying. This observation was correlated to the synthesis of new proteins needed to support early embryo development. Additional 20% of VT was consumed thereafter, mainly at the end of embryogenesis, and in this case VT is probably used as energy source to the older embryo. Altogether, these data indicate different energy sources for maternal and zygotic driven processes.     

Virulence-associated gene profiling of Streptococcus suis isolates by PCR

Definition of virulent Streptococcus suis strains is controversial. One successful approach for identification of virulent European strains is differentiation of capsular serotypes (or the corresponding cps types) and subsequent detection of virulence-associated factors, namely the extracellular factor (EF, epf), the muramidase-released protein (MRP, mrp) and the hemolysin suilysin (SLY, sly). In this work we present a novel multiplex PCR (MP-PCR) and an mrp variant PCR for identification and characterization of virulent S. suis strains. These new methods were used to identify association of disease with particular profiles of virulence-associated genes. The MP-PCR allowed identification of S. suis through detection of the housekeeping gene gdh, differentiation of four cps types (1, 2, 7 and 9), and detection of epf, mrp, sly and arcA (arginine deiminase from S. suis). Furthermore, this study describes the first PCR assay for differentiation of at least six mrp variants. Expression of the corresponding size variants of MRP was shown for four of the six mrp variants, but was undetectable for the two larger mrp variants in the particular strains investigated. The results of this study suggest that cps7 strains are associated with pneumonia and that variation of mrp is very pronounced among these strains. Gene profiles of invasive, pneumonia and carrier S. suis isolates by combination of PCR assays allowed differentiation of 24 different genotypes among cps1, 2, 7 and 9 strains. Forty-five percent of the invasive S. suis diseases investigated in this study were caused by only two of these genotypes, namely cps2/mrp+/epf+/sly+ and cps9/mrp(*)/epf-/sly+. Thus, this study demonstrates for the first time a uniform profile of the particular virulence-associated genes for the vast majority of the investigated invasive cps9 strains.     

Antinociceptive activity of the pyranocoumarin seselin in mice

Seselin an angular pyranocoumarin at dose of 0.5, 4.5 or 40.5 mg/kg inhibited the writhing response induced by acetic acid in a significant and dose-dependent manner, by 19.5%, 26.2% and 41.4%, respectively. Using the same doses, seselin elicited a significant inhibition of formalin response during the second phase (inflammatory), by 90.3%, 97.8% and 95.3%, respectively. Besides, a significant reduction of licking time was observed during the first phase (neurogenic) at the highest doses of seselin, by 34.4% and 66.9%, respectively. On the contrary, in the hot plate test no effect was observed after seselin treatment. In conclusion, seselin was able to inhibit inflammatory hyperalgesia, suggesting that this natural product possesses both important peripheral anti-inflammatory and antinociceptive properties.     

Scion-rootstock interaction affects the physiology and fruit quality of sweet cherry

Water relations, leaf gas exchange, chlorophyll a fluorescence, light canopy transmittance, leaf photosynthetic pigments and metabolites and fruit quality indices of cherry cultivars 'Burlat', 'Summit' and 'Van' growing on five rootstocks with differing size-controlling potentials that decrease in the order: Prunus avium L. > CAB 11E > Maxma 14 > Gisela 5 > Edabriz, were studied during 2002 and 2003. Rootstock genotype affected all physiological parameters. Cherry cultivars grafted on invigorating rootstocks had higher values of midday stem water potential (Psi(MD)), net CO(2) assimilation rate (A), stomatal conductance (g(s)), intercellular CO(2) concentration (C(i)) and maximum photochemical efficiency of photosystem II (PSII) (F(v)/F(m)) than cultivars grafted on dwarfing rootstocks. The Psi(MD) was positively correlated with A, g(s) and C(i). Moreover, A was positively correlated with g(s), and the slopes of the linear regression increased from invigorating to dwarfing rootstocks, indicating a stronger regulation of photosynthesis by stomatal aperture in trees on dwarfing Edabriz and Gisela 5. The effect of rootstock genotype was also statistically significant for leaf photosynthetic pigments, whereas metabolite concentrations and fruit physicochemical characteristics were more dependent on cultivar genotype. Among cultivars, 'Burlat' leaves had the lowest concentrations of photosynthetic pigments, but were richest in total soluble sugars, starch and total phenols. Compared with the other cultivars, 'Summit' had heavier fruits, independent of the rootstock. 'Burlat' cherries were less firm and had lower concentrations of soluble sugars and a lower titratable acidity than 'Van' cherries. Nevertheless, 'Van' cherries had lower lightness, chroma and hue angle, representing redder and darker cherries, compared with 'Summit' fruits. In general, Psi(MD) was positively correlated with fruit mass and A was negatively correlated with lightness and chroma. These results demonstrate that: (1) water relations and photosynthesis of sweet cherry tree are mainly influenced by the rootstock genotype; (2) different physicochemical characteristics observed in cherries of the three cultivars suggest that regulation of fruit quality was mainly dependent on the cultivar genotype, although the different size-controlling rootstocks also had a significant effect.     

Multiple phosphorylation sites confer reproducibility of the rod's single-photon responses

Although signals controlled by single molecules are expected to be inherently variable, rod photoreceptors generate reproducible responses to single absorbed photons. We show that this unexpected reproducibility-the consistency of amplitude and duration of rhodopsin activity-varies in a graded and systematic manner with the number but not the identity of phosphorylation sites on rhodopsin's C terminus. These results indicate that each phosphorylation site provides an independent step in rhodopsin deactivation and that collectively these steps tightly control rhodopsin's active lifetime. Other G protein cascades may exploit a similar mechanism to encode accurately the timing and number of receptor activation.     

Acute cattle intoxication from Nerium oleander pods

Seven outbreaks of acute intoxication from oleander (Nerium oleander) in cattle were reported in Northeast of Brazil. A total of 92 cattle were poisoned by oleander in 7 different herds; 57 animals died (67% of affected cattle). All cases reported here occurred during dry season. Two of the outbreaks resulted from offering oleander triturated and mixed with fodder. In the other cases, accidental ingestion of residual parts of oleander derived from pruning or cutting plants on grazing land were responsible. Clinical signs were diverse; the most common were locomotion disturbances, diarrhoea, depression and sudden death. Postmortem findings varied from no significant lesions to widespread haemorrhage. The presence of oleander leaves in the rumen was noted in all cases. The lack of information about the toxicity of oleanders was the main cause for the accident, which prompt us to stimulate wide divulgation of the common toxic plants.     

Extending top-down mass spectrometry to proteins with masses greater than 200 kilodaltons

For characterization of sequence and posttranslational modifications, molecular and fragment ion mass data from ionizing and dissociating a protein in the mass spectrometer are far more specific than are masses of peptides from the protein's digestion. We extend the approximately 500-residue, approximately 50-kilodalton (kD) dissociation limitation of this top-down methodology by using electrospray additives, heated vaporization, and separate noncovalent and covalent bond dissociation. This process can cleave 287 interresidue bonds in the termini of a 1314-residue (144-kD) protein, specify previously unidentified disulfide bonds between 8 of 27 cysteines in a 1714-residue (200-kD) protein, and correct sequence predictions in two proteins, one with 2153 residues (229 kD).