Development of molecular markers based on retrotransposons for the analysis of genetic variability in Moniliophthora perniciosa

Moniliophthora perniciosa is a fungus that causes witches?? broom disease (WBD) in the cacao tree (Theobroma cacao). The M. perniciosa genome contains different transposable elements; this prompted an evaluation of the use of its retrotransposons as molecular markers for population studies. The inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) techniques were used to study the variability of 70?M. perniciosa isolates from different geographic origins and biotypes. A total of 43 loci was amplified. Cluster analysis of different geographical regions of C biotype revealed two large groups in the state of Bahia, Brazil. Techniques using retrotransposon-based molecular markers showed advantages over previously used molecular techniques for the study of genetic variability in M. perniciosa.     

CO2 fertilization does not affect biomass production and nutritive value of a C4 tropical grass in short timeframe

Increased atmospheric carbon dioxide (CO₂) is a consequence of recent anthropogenic environmental changes, and few studies have evaluated its effects on tropical grasses used in Brazilian pastures, the main feed source for major part of ruminant livestock. This study evaluated forage production, chemical composition, in vitro total gas production and organic matter degradability of Brachiaria brizantha under contrasting CO₂ atmospheric conditions in a free air carbon dioxide enrichment (FACE) facility. The forage plants were sown in each of the 12 octagonal rings of the FACE facility: six under ambient atmospheric CO₂ concentration of approximately 390 μmol/mol, hereafter referred to as control (CON) plots, and other six rings enriched with pure CO₂ flux to achieve a target CO₂ concentration of 550 μmol/mol, hereafter called elevated CO₂ (eCO₂) plots. Soil samples were collected to determine carbon and nitrogen concentrations. After seventy days of sowing, a standardization cutting was performed and then at regular intervals of 21 days the forage was harvested (ten harvest dates) and forwarded to laboratorial analyses. Forage above‐ground biomass production (dry matter (DM): 6,143 vs. 6,554 kg/ha), as well as morphological characteristics (leaves: 71% vs. 68%; stem: 28% vs. 31%), chemical composition (crude protein: 162.9 vs. 161.8; neutral detergent fibre: 663.8 vs. 664.3; acid detergent fibre: 369.5 vs. 381; lignin: 60.1 vs. 64.1 g/kg DM; total C: 45.9 vs. 45.9; total N: 2.8 vs. 2.8; total S: 0.2% vs. 0.2%), organic matter in vitro degradability (573.5 vs. 585.3 g/kg), methane (5.7 vs. 4.3 ml/g DM) and total gas (128.3 vs. 94.5 ml/g DM) production did not differ significantly between CON and eCO₂ treatments (p > .05). The results indicated that at least under short‐term enrichment, B. brizantha was not affected by eCO₂.     

Isolation and characterization of a new hydroxytyrosol derivative from olive (Olea europaea) leaves

A new secoiridoid compound was isolated from the leaves of Olea europaea. This compound, not previously identified, is the bis methylacetal of oleuropein aglycone, the 3,4-dihydroxyphenylethyl [(2,6-dimethoxy-3-ethylidene)-tetrahydropyran-4-yl]acetate (3,4-DHPEA-DETA), and was found in different olive cultivar phenolic extracts as one of the major secoiridoid components. This compound was shown to be easily transformed in acidic aqueous media into 3,4-DHPEA-EDA, the major polyphenolic compound found in olive oil, and permitted us to increase the yield of 3,4-DHPEA-EDA isolation from the olive leaf extract. The antiradical activity of this new compound, evaluated by scavenging of 2,2-diphenyl-1-picrylhydrazyl radicals, was much higher than the one found for 3,4-DHPEA-EDA or alpha-tocopherol. Results also call to attention the need for a careful identification of compounds by HPLC-MS, usually performed in acidic conditions.     

Structural Analysis of Tillandsia usneoides L. Exposed to Air Pollutants in São Paulo City–Brazil

Bioindicators plants are important for the evaluation of air quality and Tillandsia usneoides L., an atmospheric epiphyte bromeliad, has been used for this purpose. The present study aims at evaluate the structural pattern of the leaf of this species when exposed to urban air pollutants, and determining whether the leaves present structural parameters that could be used as indicators of such pollutants. Samples of T. usneoides were exposed in São Paulo, the biggest city of Brazil, for 8, 16 and 24 weeks, and compared with others kept in a rural area. The urban pollution of São Paulo affected the structure of the leaves of T. usneoides causing alterations, especially in the scales, density of stomata and epidermis thickness. Qualitative alterations in the mesophyll were not observed in plants exposed at the polluted sites. These structural characteristics of T. usneoides seem to account for its high tolerance to heavy metal accumulation. The percentage of anomalous scales may potentially be used as an alternative bioindicator parameter.     

A New Bioassay Platform Design for the Discovery of Small Molecules with Anticancer Immunotherapeutic Activity

Immunotherapy takes advantage of the immune system to prevent, control, and eliminate neoplastic cells. The research in the field has already led to major breakthroughs to treat cancer. In this work, we describe a platform that integrates in vitro bioassays to test the immune response and direct antitumor effects for the preclinical discovery of anticancer candidates. The platform relies on the use of dendritic cells that are professional antigen-presenting cells (APC) able to activate T cells and trigger a primary adaptive immune response. The experimental procedure is based on two phenotypic assays for the selection of chemical leads by both a panel of nine tumor cell lines and growth factor-dependent immature mouse dendritic cells (D1). The positive hits are then validated by a secondary test on human monocyte-derived dendritic cells (MoDCs). The aim of this approach is the selection of potential immunotherapeutic small molecules from natural extracts or chemical libraries.     

Analysis of world strains of Anaplasma marginale using major surface protein 1a repeat sequences

Anaplasma marginale is a tick-borne pathogen of cattle that causes the disease bovine anaplasmosis worldwide. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. MSP1a is involved in the adhesion and transmission of A. marginale by ticks and varies among geographic strains in the number and sequence of amino-terminal tandem repeats. The aim of this study was to characterize the genetic diversity of A. marginale strains collected from countries in North and South America, Europe, Asia, Africa and Australia, inclusive of all continents. In this study, we characterized 131 strains of A. marginale using 79 MSP1a repeat sequences. These results corroborated the genetic heterogeneity of A. marginale strains in endemic regions worldwide. The phylogenetic analyses of MSP1a repeat sequences did not result in clusters according to the geographic origin of A. marginale strains but provided phylogeographic information. Seventy-eight percent of the MSP1a repeat sequences were present in strains from a single geographic region. Strong (> or =80%) support was found for clusters containing sequences from Italian, Spanish, Chinese, Argentinean and South American strains. The phylogenetic analyses of MSP1a repeat sequences suggested tick-pathogen co-evolution and provided evidence of multiple introductions of A. marginale strains from various geographic locations worldwide. These results contribute to the understanding of the genetic diversity and evolution of A. marginale and tick-pathogen interactions.     

Effects of Marine Toxins on the Reproduction and Early Stages Development of Aquatic Organisms

Marine organisms, and specially phytoplankton species, are able to produce a diverse array of toxic compounds that are not yet fully understood in terms of their main targets and biological function. Toxins such as saxitoxins, tetrodotoxin, palytoxin, nodularin, okadaic acid, domoic acid, may be produced in large amounts by dinoflagellates, cyanobacteria, bacteria and diatoms and accumulate in vectors that transfer the toxin along food chains. These may affect top predator organisms, including human populations, leading in some cases to death. Nevertheless, these toxins may also affect the reproduction of aquatic organisms that may be in contact with the toxins, either by decreasing the amount or quality of gametes or by affecting embryonic development. Adults of some species may be insensitive to toxins but early stages are more prone to intoxication because they lack effective enzymatic systems to detoxify the toxins and are more exposed to the toxins due to a higher metabolic growth rate. In this paper we review the current knowledge on the effects of some of the most common marine toxins on the reproduction and development of early stages of some organisms.     

Effects of anthropogenic noise as a source of acoustic stress in wild populations of Hippocampus guttulatus in the Ria Formosa,south Portugal

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