First isolation of Pseudogymnoascus destructans in bats from Portugal

The psychrophilic fungus Pseudogymnoascus destructans (formerly known as Geomyces destructans) is considered the etiological agent of white-nose disease (WND), an emerging disease which affects bats during their hibernation period. This disease is clinically characterized by the growth of a white fungus on muzzle, ears, and wings’ membranes of affected bats. This infection caused the death of several million bats in North America. Conversely, European bats show no evidence of significant mortality occurrences associated with P. destructans colonization. This fungus has been isolated from bats in at least 15 European countries since 2008, but was never before reported in the Iberian Peninsula. This study describes the first case report of P. destructans colonization in bats from Portugal. We isolated P. destructans from three hibernating Myotis blythii (lesser mouse-eared bat) with visual signs of P. destructans colonization, during a routine visit to a mine located in the Trás-os-Montes region, Northern Portugal. M. blythii is one of the rarest bat species in Europe, classified as critically endangered in Portugal. P. destructans was obtained from at least three different parts of the body of each specimen analyzed. The identification of the respective fungal isolates was based on the macroscopic and microscopic characterization of the cultures and confirmed by PCR-based analysis. All nucleotide sequences obtained showed 100 % identity with previous data reported for P. destructans. This new finding improves the current knowledge about the European distribution of P. destructans, which is of great interest for forthcoming studies on the fungus dispersion and impact among bat populations at regional and/or global level.     

Effects of artichoke (Cynara scolymus) leaf and bloom head extracts on chemically induced DNA lesions in Drosophila melanogaster

The genotoxicity of bloom head (BHE) and leaf (LE) extracts from artichoke (Cynara scolymus L.), and their ability to modulate the mutagenicity and recombinogenicity of two alkylating agents (ethyl methanesulfonate – EMS and mitomycin C – MMC) and the intercalating agent bleomycin (BLM), were examined using the somatic mutation and recombination test (SMART) in Drosophila melanogaster. Neither the mutagenicity nor the recombinogenicity of BLM or MMC was modified by co- or post-treatment with BHE or LE. In contrast, co-treatment with BHE significantly enhanced the EMS-induced genotoxicity involving mutagenic and/or recombinant events. Co-treatment with LE did not alter the genotoxicity of EMS whereas post-treatment with the highest dose of LE significantly increased this genotoxicity. This enhancement included a synergistic increase restricted to somatic recombination. These results show that artichoke extracts promote homologous recombination in proliferative cells of D. melanogaster.     

Generalized Allee effect model

The Allee effect consists of a positive correlation between very small population size and fitness. Offering a new view point on the weak and strong demographic Allee effect, we propose to combine them with the Richards growth model. In particular, a peculiar manifestation of the Allee effect is analytically predicted and still not validated by experiments. Model validation with ecological data is presented for some special situations.     

Floristic variation in ecotonal areas: Patterns,determinants and biogeographic origins of subtropical forests in South America

We present the first quantification of forest community composition and its relationship with environmental factors in South American subtropical Atlantic Forests. In this region, rain, seasonally dry and mixed forests form an ecotonal zone near the parallel of latitude 30°S. To investigate how well current knowledge on climatic effects and biogeographic distribution apply to subtropical ecotones, we tested the following expectations: (i) there is a floristic longitudinal gradient correlated to altitudinal and climatic gradients; (ii) climatic variables are more important than soil factors in shaping floristic composition; and (iii) there are three floristic regions in the southernmost limit of the Atlantic Forest biome that are expected to be distinct in composition, structure and biogeographical origin. We examined floristic composition and its relationship with environmental factors across 52 1‐ha permanent study areas in subtropical Brazil, containing in total 269 tree species ≥ 9.5 dbh (diameter at breast height). Climatic data, related to rainfall seasonality and temperature, as well as soil properties, were compiled from published sources or global data banks. Expectations one and two were confirmed, but expectation three was only partially met. Hierarchical cluster analysis divided the southernmost Atlantic Forests into four major groups (Rain, Seasonally Dry, Western Mixed and Eastern Mixed Forests). Overall, the tested environmental variables differed significantly among the four regions. Using indicator species analysis, we distinguished 46 indicator species, which had significant environmental preferences for one floristic region. These species can be used as indicators of environmental conditions or to determine to which floristic region a certain forest belongs. Biogeographic distributions differed between floristic groups, supporting the interpretation that Eastern Mixed Forests are relict forests of a temperate forest of Andean origin that occurred during colder palaeoclimates. Western Mixed Forests represent the main floristic ecotone between Seasonally dry and Eastern Mixed Forests.     

Balance between the two kinin receptors in the progression of experimental focal and segmental glomerulosclerosis in mice

Focal and segmental glomerulosclerosis (FSGS) is one of the most important renal diseases related to end-stage renal failure. Bradykinin has been implicated in the pathogenesis of renal inflammation, whereas the role of its receptor 2 (B2RBK; also known as BDKRB2) in FSGS has not been studied. FSGS was induced in wild-type and B2RBK-knockout mice by a single intravenous injection of Adriamycin (ADM). In order to further modulate the kinin receptors, the animals were also treated with the B2RBK antagonist HOE-140 and the B1RBK antagonist DALBK. Here, we show that the blockage of B2RBK with HOE-140 protects mice from the development of FSGS, including podocyte foot process effacement and the re-establishment of slit-diaphragm-related proteins. However, B2RBK-knockout mice were not protected from FSGS. These opposite results were due to B1RBK expression. B1RBK was upregulated after the injection of ADM and this upregulation was exacerbated in B2RBK-knockout animals. Furthermore, treatment with HOE-140 downregulated the B1RBK receptor. The blockage of B1RBK in B2RBK-knockout animals promoted FSGS regression, with a less-inflammatory phenotype. These results indicate a deleterious role of both kinin receptors in an FSGS model and suggest a possible cross-talk between them in the progression of disease.KEY WORDS: Focal and segmental glomerulosclerosis, Bradykinin receptors, Inflammation, Podocyte, Fibrosis     

The HIV-Brazil Cohort Study: Design,Methods and Participant Characteristics

Background

The HIV-Brazil Cohort Study was established to analyze the effectiveness of combination antiretroviral therapy (cART) and the impact of this treatment on morbidity, quality of life (QOL) and mortality. The study design, patients’ profiles and characteristics of cART initiation between 2003 and 2010 were described.

Methodology/Principal Findings

Since 2003, the HIV-Brazil Cohort has been following HIV-infected adults receiving cART at 26 public health care facilities, using routine clinical care data and self-reported QOL questionnaires. When not otherwise available, data are obtained from national information systems. The main outcomes of interest are diseases related or unrelated to HIV; suppression of viral replication; adverse events; virological, clinical and immunological failures; changes in the cART; and mortality. For the 5,061 patients who started cART between 2003 and 2010, the median follow-up time was 4.1 years (IQR 2.2–5.9 years) with an 83.4% retention rate. Patient profiles were characterized by a predominance of men (male/female ratio 1.7∶1), with a mean age of 36.9 years (SD 9.9 years); 55.2% had been infected with HIV via heterosexual contact. The majority of patients (53.4%) initiated cART with a CD4⁺ T-cell count ≤200 cells/mm³. The medications most often used in the various treatment regimens were efavirenz (59.7%) and lopinavir/ritonavir (18.2%). The proportion of individuals achieving viral suppression within the first 12 months of cART use was 77.4% (95% CI 76.1–78.6). Nearly half (45.4%) of the patients presented HIV-related clinical manifestations after starting cART, and the AIDS mortality rate was 13.9 per 1,000 person-years.

Conclusions/Significance

Results from cART use in the daily practice of health services remain relatively unknown in low- and middle-income countries, and studies with the characteristics of the HIV-Brazil Cohort contribute to minimizing these shortcomings, given its scope and patient profile, which is similar to that of the AIDS epidemic in the country.     

Integrating Multibeam Backscatter Angular Response,Mosaic and Bathymetry Data for Benthic Habitat Mapping

Multibeam echosounders (MBES) are increasingly becoming the tool of choice for marine habitat mapping applications. In turn, the rapid expansion of habitat mapping studies has resulted in a need for automated classification techniques to efficiently map benthic habitats, assess confidence in model outputs, and evaluate the importance of variables driving the patterns observed. The benthic habitat characterisation process often involves the analysis of MBES bathymetry, backscatter mosaic or angular response with observation data providing ground truth. However, studies that make use of the full range of MBES outputs within a single classification process are limited. We present an approach that integrates backscatter angular response with MBES bathymetry, backscatter mosaic and their derivatives in a classification process using a Random Forests (RF) machine-learning algorithm to predict the distribution of benthic biological habitats. This approach includes a method of deriving statistical features from backscatter angular response curves created from MBES data collated within homogeneous regions of a backscatter mosaic. Using the RF algorithm we assess the relative importance of each variable in order to optimise the classification process and simplify models applied. The results showed that the inclusion of the angular response features in the classification process improved the accuracy of the final habitat maps from 88.5% to 93.6%. The RF algorithm identified bathymetry and the angular response mean as the two most important predictors. However, the highest classification rates were only obtained after incorporating additional features derived from bathymetry and the backscatter mosaic. The angular response features were found to be more important to the classification process compared to the backscatter mosaic features. This analysis indicates that integrating angular response information with bathymetry and the backscatter mosaic, along with their derivatives, constitutes an important improvement for studying the distribution of benthic habitats, which is necessary for effective marine spatial planning and resource management.     

Structure-Activity Relationship of Benzophenanthridine Alkaloids from Zanthoxylum rhoifolium Having Antimicrobial Activity

Zanthoxylum rhoifolium (Rutaceae) is a plant alkaloid that grows in South America and has been used in Brazilian traditional medicine for the treatment of different health problems. The present study was designed to evaluate the antimicrobial activity of the steam bark crude methanol extract, fractions, and pure alkaloids of Z. rhoifolium. Its stem bark extracts exhibited a broad spectrum of antimicrobial activity, ranging from 12.5 to 100 µg/mL using bioautography method, and from 125 to 500 µg/mL in the microdilution bioassay. From the dichloromethane basic fraction, three furoquinoline alkaloids (1–3), and nine benzophenanthridine alkaloids (4–12) were isolated and the antimicrobial activity of the benzophenanthridine alkaloids is discussed in terms of structure-activity relationships. The alkaloid with the widest spectrum of activity was chelerythrine (10), followed by avicine (12) and dihydrochelerythrine (4). The minimal inhibitory concentrations of chelerythrine, of 1.50 µg/mL for all bacteria tested, and between 3.12 and 6.25 µg/mL for the yeast tested, show this compound to be a more powerful antimicrobial agent when compared with the other active alkaloids isolated from Z. rhoifolium. To verify the potential importance of the methylenedioxy group (ring A) of these alkaloids, chelerythrine was selected to represent the remainder of the benzophenanthridine alkaloids isolated in this work and was subjected to a demethylation reaction giving derivative 14. Compared to chelerythrine, the derivative (14) was less active against the tested bacteria and fungi. Kinetic measurements of the bacteriolytic activities of chelerythrine against the bacteria Bacillus subtilis (Gram-positive) and Escherichia coli (Gram-negative) were determined by optical density based on real time assay, suggesting that its mechanism of action is not bacteriolytic. The present study did not detect hemolytic effects of chelerythrine on erythrocytes and found a protective effect considering the decrease in TBARS and AOPP (advanced oxidized protein products) levels when compared to the control group.