Structural studies with crotoxin B from Crotalus durissus collilineatus venom suggest a heterodimeric assembly formed by two new isoforms.

In accidents involving Crotalus snakes, the crotoxin complex (CTX) plays lethal action due to its neurotoxic activity. On the other hand, CTX have potential biotechnological application due to its anti-tumoral, anti-inflammatory, antimicrobial, analgesic and immunomodulatory properties. CTX is a heterodimer composed of Crotoxin A (CA or crotapotin), the acidic nontoxic and non-enzymatic component and; Crotoxin B (CB), a basic, toxic and catalytic PLA2. Currently, there are two classes of CTX isoforms, whose differences in their biological activities have been attributed to features presented in CB isoforms. Here, we present the crystal structure of CB isolated from the Crotalus durissus collilineatus venom. It amino acid sequence was assigned using the SEQUENCE SLIDER software, which revealed that the crystal structure is a heterodimer composed of two new CB isoforms (colCB-A and colCB-B). Bioinformatic and biophysical analyses showed that the toxin forms a tetrameric assembly in solution similar to CB from Crotalus durissus terrificus venom, despite some differences observed at the dimeric interface. By the previously proposed classification, the colCB-B presents features of the class I isoforms while colCB-A cannot be classified into classes I and II based on its amino acid sequence. Due to similar features observed for other CB isoforms found in the NCBI database and the results obtained for colCB-A, we suggest that there are more than two classes of CTX and CB isoforms in crotalic venoms.

Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis.

Single-particle cryo-electron microscopy (cryo-EM SPA) has recently emerged as an exceptionally well-suited technique for determining the structure of membrane proteins (MPs). Indeed, in recent years, huge increase in the number of MPs solved via cryo-EM SPA at a resolution better than 3.0 Å in the Protein Data Bank (PDB) has been observed. However, sample preparation remains a significant challenge in the field. Here, we evaluated the MPs solved using cryo-EM SPA deposited in the PDB in the last two years at a resolution below 3.0 Å. The most critical parameters for sample preparation are as follows: (i) the surfactant used for protein extraction from the membrane, (ii) the surfactant, amphiphiles, nanodiscs or other molecules present in the vitrification step, (iii) the vitrification method employed, and (iv) the type of grids used. The aim is not to provide a definitive answer on the optimal sample conditions for cryo-EM SPA of MPs but rather assess the current trends in the MP structural biology community towards obtaining high-resolution cryo-EM structures.

Modelling level I Axillary Lymph Nodes depth for Microwave Imaging.

PURPOSE: Patient-specific information on the depth of Axillary Lymph Nodes (ALNs) is important for the development of new diagnostic imaging technologies, e.g. Microwave Imaging (MWI), aiming to assess the diagnosis of ALNs during breast cancer staging. Studies about ALNs depth have been presented for treatment planning, but they lack information on sample size and usability of the data to infer the depth of ALNs. The aim of this study was to create a mathematical model that can be used to predict a depth interval where level I ALNs are likely to be located. METHODS: We extracted biometric features of 98 patients who underwent breast Magnetic Resonance Imaging (MRI) to train two types of regression models. We then tested different combination of features to predict ALNs depth and found the best predictor. The final prediction models were then implemented in an algorithm used for MWI and tested with anthropomorphic phantoms of the axillary region. RESULTS: Body Mass Index (BMI) was the feature with best performance to predict ALNs depth with coefficient of determination (R2) ranging from 0.49 to 0.55 and Root Mean Squared Error (RMSE) ranging from 0.68 to 0.91 cm. The proposed model showed satisfactory results in microwave images of patients with different BMIs. CONCLUSIONS: The presented results contribute to the development of reconstruction algorithms for new imaging technologies and to the assessment of ALNs in other medical applications.

Cryo-electron microscopy unveils unique structural features of the human Kir2.1 channel.

We present the first structure of the human Kir2.1 channel containing both transmembrane domain (TMD) and cytoplasmic domain (CTD). Kir2.1 channels are strongly inward-rectifying potassium channels that play a key role in maintaining resting membrane potential. Their gating is modulated by phosphatidylinositol 4,5-bisphosphate (PIP2). Genetically inherited defects in Kir2.1 channels are responsible for several rare human diseases, including Andersen's syndrome. The structural analysis (cryo-electron microscopy), surface plasmon resonance, and electrophysiological experiments revealed a well-connected network of interactions between the PIP2-binding site and the G-loop through residues R312 and H221. In addition, molecular dynamics simulations and normal mode analysis showed the intrinsic tendency of the CTD to tether to the TMD and a movement of the secondary anionic binding site to the membrane even without PIP2. Our results revealed structural features unique to human Kir2.1 and provided insights into the connection between G-loop and gating and the pathological mechanisms associated with this channel.

Increase in Hepatitis A Cases Linked to Imported Strains to Rio de Janeiro, Brazil: A Cross-Sectional Study.

This study aims to evaluate the epidemiological and molecular features associated with HAV transmission in adults in Rio de Janeiro during a period of increased registered cases of HAV (2017-2018). Socio-epidemiological data and serum samples from anti-HAV IgM+ individuals were obtained. HAV RNA was RT-PCR amplified and sequenced for further phylogenetic and phylogeographic analyses. From fifty-two HAV IgM+ individuals, most were men (78.85%; p = 0.024), aged 20-30 years old (84.61%; p < 0.001), resided in the Rio de Janeiro north zone (31/52; 59.62%; p = 0.001), and are men who have sex with men (MSM) (57.69%; p = 0.002). Sexual practices were more frequent (96%) than others risk factors (food-borne (44%), water-borne (42.31%), and parenteral (34.62%)). Individuals who traveled to endemic regions had a 7.19-fold (1.93-36.04; p < 0.01) increased risk of HAV. Phylogenetic analysis revealed four distinct clades of subgenotype IA, three of them comprised sequences from European/Asian MSM outbreaks and one from Brazilian endemic strains. Bayesian Inference showed that the imported strains were introduced to Brazil during large mass sportive events. Sexual orientation and sexual practices may play a role in acquiring HAV infection. Public policies targeting key populations must be implemented to prevent further dissemination of HAV and other STIs.

Development of MRI-based axillary numerical models and estimation of axillary lymph node dielectric properties for microwave imaging.

PURPOSE: Microwave imaging (MWI) has been studied as a complementary imaging modality to improve sensitivity and specificity of diagnosis of axillary lymph nodes (ALNs), which can be metastasized by breast cancer. The feasibility of such a system is based on the dielectric contrast between healthy and metastasized ALNs. However, reliable information such as anatomically realistic numerical models and matching dielectric properties of the axillary region and ALNs, which are crucial to develop MWI systems, are still limited in the literature. The purpose of this work is to develop a methodology to infer dielectric properties of structures from magnetic resonance imaging (MRI), in particular, ALNs. We further use this methodology, which is tailored for structures farther away from MR coils, to create MRI-based numerical models of the axillary region and share them with the scientific community, through an open-access repository. METHODS: We use a dataset of breast MRI scans of 40 patients, 15 of them with metastasized ALNs. We apply image processing techniques to minimize the artifacts in MR images and segment the tissues of interest. The background, lung cavity, and skin are segmented using thresholding techniques and the remaining tissues are segmented using a K-means clustering algorithm. The ALNs are segmented combining the clustering results of two MRI sequences. The performance of this methodology was evaluated using qualitative criteria. We then apply a piecewise linear interpolation between voxel signal intensities and known dielectric properties, which allow us to create dielectric property maps within an MRI and consequently infer ALN properties. Finally, we compare healthy and metastasized ALN dielectric properties within and between patients, and we create an open-access repository of numerical axillary region numerical models which can be used for electromagnetic simulations. RESULTS: The proposed methodology allowed creating anatomically realistic models of the axillary region, segmenting 80 ALNs and analyzing the corresponding dielectric properties. The estimated relative permittivity of those ALNs ranged from 16.6 to 49.3 at 5 GHz. We observe there is a high variability of dielectric properties of ALNs, which can be mainly related to the ALN size and, consequently, its composition. We verified an average dielectric contrast of 29% between healthy and metastasized ALNs. Our repository comprises 10 numerical models of the axillary region, from five patients, with variable number of metastasized ALNs and body mass index. CONCLUSIONS: The observed contrast between healthy and metastasized ALNs is a good indicator for the feasibility of a MWI system aiming to diagnose ALNs. This paper presents new contributions regarding anatomical modeling and dielectric properties' characterization, in particular for axillary region applications.

Gallic acid anti-myotoxic activity and mechanism of action, a snake venom phospholipase A2 toxin inhibitor, isolated from the medicinal plant Anacardium humile.

Snakebite envenoming is the cause of an ongoing health crisis in several regions of the world, particularly in tropical and neotropical countries. This scenario creates an urgent necessity for new practical solutions to address the limitations of current therapies. The current study investigated the isolation, phytochemical characterization, and myotoxicity inhibition mechanism of gallic acid (GA), a myotoxin inhibitor obtained from Anacardium humile. The identification and isolation of GA was achieved by employing analytical chromatographic separation, which exhibited a compound with retention time and nuclear magnetic resonance spectra compatible with GA's commercial standard and data from the literature. GA alone was able to inhibit the myotoxic activity induced by the crude venom of Bothrops jararacussu and its two main myotoxins, BthTX-I and BthTX-II. Circular dichroism (CD), fluorescence spectroscopy (FS), dynamic light scattering (DLS), and interaction studies by molecular docking suggested that GA forms a complex with BthTX-I and II. Surface plasmon resonance (SPR) kinetics assays showed that GA has a high affinity for BthTX-I with a KD of 9.146 × 10-7 M. Taken together, the two-state reaction mode of GA binding to BthTX-I, and CD, FS and DLS assays, suggest that GA is able to induce oligomerization and secondary structure changes for BthTX-I and -II. GA and other tannins have been shown to be effective inhibitors of snake venoms' toxic effects, and herein we demonstrated GA's ability to bind to and inhibit a snake venom PLA2, thus proposing a new mechanism of PLA2 inhibition, and presenting more evidence of GA's potential as an antivenom compound.

RPA-1 from Leishmania sp.: Recombinant Protein Expression and Purification, Molecular Modeling, and Molecular Dynamics Simulations Protocols.

RPA is a conserved heterotrimeric complex and the major single-stranded DNA (ssDNA)-binding protein heterotrimeric complex, which in eukaryotes is formed by the RPA-1, RPA-2, and RPA-3 subunits. The main structural feature of RPA is the presence of the oligonucleotide/oligosaccharide-binding fold (OB-fold) domains, responsible for ssDNA binding and protein:protein interactions. Among the RPA subunits, RPA-1 bears three of the four OB folds involved with RPA-ssDNA binding, although in some organisms RPA-2 can also bind ssDNA. The OB-fold domains are also present in telomere end-binding proteins (TEBP), essential for chromosome end protection. RPA-1 from Leishmania sp., as well as RPA-1 from trypanosomatids, a group of early-divergent protozoa, shows some structural differences compared to higher eukaryote RPA-1. Also, RPA-1 from Leishmania sp., similar to TEBPs, may exert telomeric protective functions. Remarkably, different pieces of evidence have pointed out that trypanosomatids may not have OB fold-containing TEBPs. Moreover, recent data indicate that trypanosomatid RPA-1 may be considered a TEBP since it shares with TEBPs conserved functional and structural features. However, it is still unknown whether the RPA-1 protective telomeric role is exclusive to trypanosomatids or is also present in other primitive eukaryotes. Here, we describe a protocol to obtain highly purified and biologically active Leishmania amazonensis recombinant RPA-1, and to perform molecular modeling and molecular dynamics simulations methods which could be probably applied to functional and structural studies of homologous proteins in other primitive eukaryotes.

Evaluation of a Dielectric-Only Transmitarray for Generating Multi-Focusing Near-Field Spots Using a Cluster of Feeds in the Ka-Band.

A transmitarray antenna is evaluated to generate a multi-focusing spot area in the Fresnel region of the antenna in the Ka-band. The antenna is designed to focus the radiated field at a certain point using a central feeding configuration. The number of feeds is increased to create as many focusing spots as feeds. The feeds are placed along an arc defined in the principal planes of the transmitarray, radiating independent near-field spots and providing a solution with a wide-angle spot scanning without an antenna displacement and a high isolation between feeds. To validate this concept, a transmitarray based on dielectric-only cells is designed and simulated under full-wave conditions. Then, this design is manufactured using a 3D printing technique, and the prototype is measured in a planar acquisition range. Measurements are performed for different feed positions in order to validate the multi-focusing capability of the antenna. Measurements and simulations show a high agreement and validate the proposed design technique.

Characterization and Design of Photovoltaic Solar Cells That Absorb Ultraviolet, Visible and Infrared Light.

The world is witnessing a tide of change in the photovoltaic industry like never before; we are far from the solar cells of ten years ago that only had 15-18% efficiency. More and more, multi-junction technologies seem to be the future for photovoltaics, with these technologies already hitting the mark of 30% under 1-sun. This work focuses especially on a state-of-the-art triple-junction solar cell, the GaInP/GaInAs/Ge lattice-matched, that is currently being used in most satellites and concentrator photovoltaic systems. The three subcells are first analyzed individually and then the whole cell is put together and simulated. The typical figures-of-merit are extracted; all the I-V curves obtained are presented, along with the external quantum efficiencies. A study on how temperature affects the cell was done, given its relevance when talking about space applications. An overall optimization of the cell is also elaborated; the cell's thickness and doping are changed so that maximum efficiency can be reached. For a better understanding of how varying both these properties affect efficiency, graphic 3D plots were computed based on the obtained results. Considering this optimization, an improvement of 0.2343% on the cell's efficiency is obtained.

Seroprevalence of human alphaherpesvirus 1 and 2 among pregnant women infected or uninfected with Zika virus from Rio de Janeiro, Brazil.

Pregnant women are an important group to be monitored for infection due to the risk of transmitting infections to their babies. Both herpes simples virus (HSV) and Zika virus (ZIKV) are neurotropic viruses that can be transmitted congenitally. In this study, the prevalence and risk factors of HSV among Zika-positive and -negative pregnant women from Rio de Janeiro, Brazil, were evaluated and compared. About 167 serum samples included in our study were from pregnant women with ZIKV infection symptoms, who were attended to in different hospitals in Rio de Janeiro between November 2015 to February 2016. Blood samples collected from 167 pregnant women were used for this study. The presence of HSV antibodies and viremia were evaluated by commercial ELISA and quantitative real-time polymerase chain reaction analyses, respectively. The data obtained from medical records were statistically analyzed. The HSV-1 and HSV-2 prevalence among pregnant women was 80.2% and 12.5% for Zika-positive women and 84.5% and 5.6% for Zika-negative women, respectively. None of the pregnant women exhibited HSV viremia. Age, trimester of gestation, and skin color were associated with HSV-1 and HSV-2 prevalence among the groups studied. HSV-2 was more prevalent in Zika-positive pregnant women than in Zika-negative pregnant women, and this simultaneous infection should be better investigated in future studies.

Occurrence of euchromatic B chromosomes in natural populations of Moenkhausia bonita and M. forestii (Pisces: Characidae)

Moenkhausia is a highly specious genus among the Characidae, composed of 96 valid species. Only twelve species have a known karyotype. Thus, here are presented the first cytogenetic data of two allopatric populations of Moenkhausia bonita and one of M. forestii, both belonging to the upper Paraná River basin (PR) with discussion on the evolutionary and cytotaxonomic aspects of the genus. The two species presented 2n = 50 chromosomes but different karyotype formulas and occurrence of 1-2 B chromosomes. These elements are small metacentrics in M. bonita and small acrocentrics in M. forestii. In both species, B chromosomes were euchromatic. Ag-NOR sites were found in pair 3 (metacentric), coinciding with fluorescent in situ hybridization (FISH) by the 18S rDNA probe in both species. However, the species differed in terms of the number and position of 5S rDNA sites. Heterochromatic blocks, mapped in M. bonita showed the least amount of heterochromatin in the terminal and pericentromeric regions, while the M. forestii karyotype revealed a greater amount of interstitial heterochromatic blocks. The karyotype distinctions between the two species, including the morphology of B chromosomes, may contribute as a reference in the taxonomic studies in this group.(AU)

Moenkhausia é um gênero altamente especioso dentre os Characidae, composto por 96 espécies válidas, mas apenas doze espécies têm seus cariótipos conhecidos. Portanto, são apresentados aqui os primeiros dados citogenéticos de duas populações alopátricas de Moenkhausia bonita e uma de M. forestii, ambas pertencentes à bacia do alto rio Paraná (PR), com uma ampla discussão sobre os aspectos evolutivos e citotaxonômicos do gênero. As duas espécies apresentaram 2n = 50 cromossomos, mas diferentes fórmulas cariotípicas e ocorrência de 1-2 cromossomos B. Esses elementos são pequenos metacêntricos em M. bonita e acrocêntricos pequenos em M. forestii. Em ambas as espécies, os cromossomos B apresentaram-se eucromáticos. Sítios Ag-NOR foram encontrados no par 3 (metacêntrico), coincidindo com a hibridização fluorescente in situ (FISH) pela sonda 18S rDNA em ambas as espécies. No entanto, as espécies diferiram em termos de número e posição dos sítios de 5S rDNA. Blocos heterocromáticos mapeados em M. bonita revelaram pequena quantidade de heterocromatina nas regiões terminal e pericentromérica, enquanto o cariótipo de M. forestii revelou uma maior quantidade de blocos heterocromáticos intersticiais. As distinções cariotípicas entre as duas espécies, incluindo a morfologia dos cromossomos B, podem contribuir como uma referência em estudos taxonômicos neste grupo.(AU)

Influence of an aluminium concentrator corrosion on the output characteristic of a photovoltaic system.

The climate changes observed over the last decades have been promoting a massive transformation on the energy sector, that is still, in truth highly dependent on fossil fuels. Renewable energies are a plausible alternative, because they have lower emissions of toxic gases in comparison with non-renewable ones. In the group of renewable energies, solar technology has the biggest overall potential, mainly because it is cheap and easy to set. Several solar technologies allow to equip their photovoltaic panels with concentrators, mostly to increase the output power and possibly their efficiency. However, some problems related to the use of concentrators have to be dealt in order to improve the entire photovoltaic system performance. One of these issues is the corrosion of the concentrators, leading to a premature ageing and, consequently an increase in maintenance costs. This problem is going to be analysed in this paper, presenting some simulation from a ray traicing software and also some experimental results, from our own laboratory experiences. The used software allows to trace the solar rays of the concentrator, in order to assess the effect of the defects caused by corrosion due to the ambient circumstances. After it, experimental results will help to analyse this effect and to prove simulation ones.

Study of the high prevalence and cardiovascular risk factors: students aged 11 to 16 years from Caceres-Spain and Paranavaí-Brazil / Estudio de alta prevalencia de factores de riesgo cardiovasculares: estudiantes de 11 a 16 años de Cáceres-España y Paranavaí-Brasil

OBJECTIVE: to investigate the association of risk factors for the development of cardiovascular diseases in students from 11 to 16 years old in different contexts. MATERIAL AND METHOD: Sample composed by students of Cáceres - Spain (n = 165) and Paranavaí - Brazil (n = 237). Body Mass Index, level of physical activity, blood pressure, total cholesterol, low- and high-density lipoproteins, triglycerides and fasting blood glucose were analyzed. For the analysis of variables, the Kolmogorov-Smirnov tests, Student t, Mann-Whitney U, Chi-square, and Odds Ratio were used, with a 95% confidence interval, a value of p <0.05 was considered statistically significant. RESULTS: Spanish students have higher average values of age, physical activity level, obesity in general, low- and high-density lipoproteins and fasting blood glucose (p <0.05). Brazilians had a greater accumulation of risk factors compared to the Spanish, specifically only 8.5% of Brazilians are exempt from RF compared to 28.2% of Spanish. It is observed that there is a higher prevalence of two RFs (G-BRA: 40.7% vs. G-ESP: 24.2%); and three or more RF (G-BRA 27.0% vs. G-ESP: 13.7%), considering a value of p <0.001. Being more active was associated with HDL levels among Brazilians. Although Spanish students had a higher prevalence of general obesity, they were more active. CONCLUSION: Spanish students showed better results in physical activity levels, fasting glucose concentration, high and low density lipoproteins, in addition to a lower number cardiovascular risk factors, despite being mainly from public schools; with higher average age and higher general obesity prevalence, compared to Brazilians

OBJETIVO: Investigar la asociación de factores de riesgo para el desarrollo de enfermedades cardiovasculares en estudiantes de 11 a 16 años en diferentes contextos. MATERIAL Y MÉTODO: Muestra compuesta por estudiantes de Cáceres - España (n = 165) y Paranavaí - Brasil (n = 237). Se analizaron el Índice de Masa Corporal, el nivel de actividad física, la presión arterial, el colesterol total, las lipoproteínas de baja y alta densidad, los triglicéridos y la glucosa en sangre en ayunas. Para el análisis de variables, se utilizaron las pruebas de Kolmogorov-Smirnov, t de Student, U de Mann-Whitney, Chi-cuadrado y Odds Ratio, con un intervalo de confianza del 95%. Un valor de p < 0,05 fue considerado estadísticamente significativo. RESULTADOS: Los estudiantes españoles tienen valores promedio más altos de edad, nivel de actividad física, obesidad en general, lipoproteínas de baja y alta densidad y glucosa en sangre en ayunas (p < 0.05). Los brasileños tuvieron una mayor acumulación de Factores de Riesgo en comparación con los españoles, específicamente solo el 8,5% de los brasileños están exentos de Factores de Riesgo en comparación con el 28,2% de los españoles. Se observa que existe una mayor prevalencia de dos Factores de Riesgo (G-BRA: 40,7% vs. G-ESP: 24,2%); y tres o más Factores de Riesgo (G-BRA 27,0% vs. G-ESP: 13,7%), considerando un valor de p < 0,001. Ser más activo se asoció con los niveles de lipoproteínas de alta densidad entre los brasileños. Aunque los estudiantes españoles tenían una mayor prevalencia de obesidad general, eran más activos. CONCLUSIÓN: Los estudiantes españoles mostraron mejores resultados en niveles de actividad física, concentración de glucosa en ayunas, lipoproteínas de alta y baja densidad, además de un menor número de factores de riesgo para enfermedades cardiovasculares, a pesar de que provienen en su mayoría de escuelas públicas; con mayor edad media y mayor prevalencia de obesidad general

Study of the Refraction Effects in Microwave Breast Imaging Using a Dry Setup.

Medical Microwave Imaging (MWI) has been studied as a technique to aid breast cancer diagnosis. Several different prototypes have been proposed but most of them require the use of a coupling medium between the antennas and the breast, in order to reduce skin backscattering and avoid refraction effects. The use of dry setups has been addressed and recent publications show promising results. In this paper, we assess the importance of considering refraction effects in the image reconstruction algorithms. To this end, we consider a simplified homogeneous spherical model of the breast and analytically compute the propagating rays through the air-body interface. The comparison of results considering only direct ray propagation or refracted rays shows negligible impact on the accuracy of the images for moderately high permittivity media. Thus, we may avoid the computational burden of calculating the refracted rays in convex shapes.

Development of an Anthropomorphic Phantom of the Axillary Region for Microwave Imaging Assessment.

We produced an anatomically and dielectrically realistic phantom of the axillary region to enable the experimental assessment of Axillary Lymph Node (ALN) imaging using microwave imaging technology. We segmented a thoracic Computed Tomography (CT) scan and created a computer-aided designed file containing the anatomical configuration of the axillary region. The phantom comprises five 3D-printed parts representing the main tissues of interest of the axillary region for the purpose of microwave imaging: fat, muscle, bone, ALNs, and lung. The phantom allows the experimental assessment of multiple anatomical configurations, by including ALNs of different size, shape, and number in several locations. Except for the bone mimicking organ, which is made of solid conductive polymer, we 3D-printed cavities to represent the fat, muscle, ALN, and lung and filled them with appropriate tissue-mimicking liquids. Existing studies about complex permittivity of ALNs have reported limitations. To address these, we measured the complex permittivity of both human and animal lymph nodes using the standard open-ended coaxial-probe technique, over the 0.5 GHz-8.5 GHz frequency band, thus extending current knowledge on dielectric properties of ALNs. Lastly, we numerically evaluated the effect of the polymer which constitutes the cavities of the phantom and compared it to the realistic axillary region. The results showed a maximum difference of 7 dB at 4 GHz in the electric field magnitude coupled to the tissues and a maximum of 10 dB difference in the ALN response. Our results showed that the phantom is a good representation of the axillary region and a viable tool for pre-clinical assessment of microwave imaging technology.

Isolation and structural characterization of bioactive compound from Aristolochia sprucei aqueous extract with anti-myotoxic activity.

A bioactive compound isolated from the stem extract of Aristolochia sprucei through High Performance Liquid Chromatography (HPLC) was identified via Nuclear Magnetic Resonance (NMR) as the aristolochic acid (AA). This compound showed an inhibitory effect over the myotoxic activity of Bothrops jararacussu and Bothrops asper venoms, being also effective against the indirect hemolytic activity of B. asper venom. Besides, AA also inhibited the myotoxic activity of BthTX-I and MTX-II with an efficiency greater than 60% against both myotoxins. Docking predictions revealed an interesting mechanism, through which the AA displays an interaction profile consistent with its inhibiting abilities, binding to both active and putative sites of svPLA2. Overall, the present findings indicate that AA may bind to critical regions of myotoxic Asp 49 and Lys49-PLA2s from snake venoms, highlighting the relevance of domains comprising the active and putative sites to inhibit these toxins.

The psychology of online activism and social movements: relations between online and offline collective action.

We review online activism and its relations with offline collective action. Social media facilitate online activism, particularly by documenting and collating individual experiences, community building, norm formation, and development of shared realities. In theory, online activism could hinder offline protests, but empirical evidence for slacktivism is mixed. In some contexts, online and offline action could be unrelated because people act differently online versus offline, or because people restrict their actions to one domain. However, most empirical evidence suggests that online and offline activism are positively related and intertwined (no digital dualism), because social media posts can mobilise others for offline protest. Notwithstanding this positive relationship, the internet also enhances the visibility of activism and therefore facilitates repression in repressive contexts.

A multi-approach analysis highlights the relevance of RPA-1 as a telomere end-binding protein (TEBP) in Leishmania amazonensis.

BACKGROUND: Telomeres are chromosome end structures important in the maintenance of genome homeostasis. They are replenished by the action of telomerase and associated proteins, such as the OB (oligonucleotide/oligosaccharide-binding)-fold containing telomere-end binding proteins (TEBP) which plays an essential role in telomere maintenance and protection. The nature of TEBPs is well known in higher and some primitive eukaryotes, but it remains undetermined in trypanosomatids. Previous in silico searches have shown that there are no homologs of the classical TEPBs in trypanosomatids, including Leishmania sp. However, Replication Protein A subunit 1 (RPA-1), an OB-fold containing DNA-binding protein, was found co-localized with trypanosomatids telomeres and showed a high preference for the telomeric G-rich strand. METHODS AND RESULTS: We predicted the absence of structural homologs of OB-fold containing TEBPs in the Leishmania sp. genome using structural comparisons. We demonstrated by molecular docking that the ssDNA binding mode of LaRPA-1 shares features with the higher eukaryotes POT1 and RPA-1 crystal structures ssDNA binding mode. Using fluorescence spectroscopy, protein-DNA interaction assays, and FRET, we respectively show that LaRPA-1 shares some telomeric functions with the classical TEBPs since it can bind at least one telomeric repeat, protect the telomeric G-rich DNA from 3'-5' Exonuclease I digestion, and unfold telomeric G-quadruplex. CONCLUSIONS: Our results suggest that RPA-1 emerges as a TEBP in trypanosomatids, and in this context, we present two possible evolutionary landscapes of trypanosomatids RPA-1 that could reflect upon the evolution of OB-fold containing TEBPs from all eukaryotes.

Nuclear export of replication protein A in the nonreplicative infective forms of Trypanosoma cruzi.

Replication protein A (RPA), a heterotrimeric complex, is the major single-stranded DNA binding protein in eukaryotes. Recently, we characterized RPA from Trypanosoma cruzi, showing that it is involved in DNA replication and DNA damage response in this organism. Better efficiency in differentiation from epimastigote to metacyclic trypomastigote forms was observed in TcRPA-2 subunit heterozygous knockout cells, suggesting that RPA is involved in this process. Here, we show that RPA cellular localization changes during the T. cruzi life cycle, with RPA being detected only in the cytoplasm of the metacyclic and bloodstream trypomastigotes. We also identify a nuclear export signal (NES) in the trypanosomatid RPA-2 subunit. Mutations in the negatively charged residues of RPA-2 NES impair the differentiation process, suggesting that RPA exportation affects parasite differentiation into infective forms.