A Phosphoproteomic Approach towards the Understanding of the Role of TGF-β in Trypanosoma cruzi Biology

Transforming growth factor beta (TGF-β) plays a pivotal role in Chagas disease, not only in the development of chagasic cardiomyopathy, but also in many stages of the T. cruzi life cycle and survival in the host cell environment. The intracellular signaling pathways utilized by T. cruzi to regulate these mechanisms remain unknown. To identify parasite proteins involved in the TGF-β response, we utilized a combined approach of two-dimensional gel electrophoresis (2DE) analysis and mass spectrometry (MS) protein identification. Signaling via TGF-β is dependent on events of phosphorylation, which is one of the most relevant and ubiquitous post-translational modifications for the regulation of gene expression, and especially in trypanosomatids, since they lack several transcriptional control mechanisms. Here we show a kinetic view of T. cruzi epimastigotes (Y strain) incubated with TGF-β for 1, 5, 30 and 60 minutes, which promoted a remodeling of the parasite phosphorylation network and protein expression pattern. The altered molecules are involved in a variety of cellular processes, such as proteolysis, metabolism, heat shock response, cytoskeleton arrangement, oxidative stress regulation, translation and signal transduction. A total of 75 protein spots were up- or down-regulated more than twofold after TGF-β treatment, and from these, 42 were identified by mass spectrometry, including cruzipain–the major T. cruzi papain-like cysteine proteinase that plays an important role in invasion and participates in the escape mechanisms used by the parasite to evade the host immune system. In our study, we observed that TGF-β addition favored epimastigote proliferation, corroborating 2DE data in which proteins previously described to be involved in this process were positively stimulated by TGF-β

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

18^{18}Ne production for the Beta beams project

Intense relativistic (anti)neutrino beams are an unique tool required to study fundamental properties of neutrinos such as neutrino oscillation parameters, as well as their Majorana or Dirac nature, the lepton number conservation hypothesis and the absolute neutrino mass scale. Such beams originate from acceleration of beta-decaying radioactive ions (“Beta beams”). A molten fluoride salt target has been developed for the production of the required rates of low-Q baseline isotope18Nefor the Beta beams project. The prototyped unit has been tested on-line at ISOLDE-CERN. In this contribution an overview of the prototyping and on-line tests is presented

Comparison on the production of radionuclides in 1.4 GeV proton irradiated LBE targets of different thickness

This is the first report on the inventory of radionuclides produced in 1.4 GeV proton induced reaction on Lead-Bismuth Eutectic (LBE) targets. LBE targets of 6 mm diameter and 1 to 8 mm lengths were irradiated with 1.4 GeV protons. The radionuclides ranging from Be-7 (53.12 days) to Po-207 (5.8 h) were identified in the samples with the help of time resolved gamma-ray spectroscopy. However, there is no signature of formation of At radioisotopes, which can be produced by the interaction of secondary particles, typical for thick targets

Prediction equations for maximal respiratory pressures of Brazilian adolescents

BACKGROUND: The literature emphasizes the need for studies to provide reference values and equations able to predict respiratory muscle strength of Brazilian subjects at different ages and from different regions of Brazil.OBJECTIVES:To develop prediction equations for maximal respiratory pressures (MRP) of Brazilian adolescents.METHOD: In total, 182 healthy adolescents (98 boys and 84 girls) aged between 12 and 18 years, enrolled in public and private schools in the city of Natal-RN, were evaluated using an MVD300 digital manometer (Globalmed®) according to a standardized protocol. Statistical analysis was performed using SPSS Statistics 17.0 software, with a significance level of 5%. Data normality was verified using the Kolmogorov-Smirnov test, and descriptive analysis results were expressed as the mean and standard deviation. To verify the correlation between the MRP and the independent variables (age, weight, height and sex), the Pearson correlation test was used. To obtain the prediction equations, stepwise multiple linear regression was used.RESULTS: The variables height, weight and sex were correlated to MRP. However, weight and sex explained part of the variability of MRP, and the regression analysis in this study indicated that these variables contributed significantly in predicting maximal inspiratory pressure, and only sex contributed significantly to maximal expiratory pressure.CONCLUSION: This study provides reference values and two models of prediction equations for maximal inspiratory and expiratory pressures and sets the necessary normal lower limits for the assessment of the respiratory muscle strength of Brazilian adolescents

TGF-β responsive proteins identified by MALDI TOF-TOF.

<p>pI theor = theoretical isoelectric point.</p><p>pI exp = experimental isoelectric point.</p><p>MW theor = theoretical molecular weight.</p><p>MW exp = experimental molecular weight.</p

TGF-β induces cruzipain expression in <i>T. cruzi</i> epimastigotes.

<p>Parasites treated (B) or not with TGF-β (A) for 5 minutes were immunostained for cruzipain and corresponding histograms were obtained using the ImageJ software (C and D). The quantification of the parasite areas labeled for cruzipain were determinate by the mean of RGB color and demonstrated as a bar graphic (E).</p

Bidimensional phosphoproteome of <i>T. cruzi</i> Y strain epimastigotes.

<p>Representative images show the total protein patterns of <i>T. cruzi</i> treated or not with TGF-β for 1, 5, 30 or 60 minutes. Protein extracts (100 µg) were separated on 7 cm pH 3–10NL IPG strips and 12% SDS-PAGE gels. Phosphoproteins were stained with Pro-Q Diamond (shown in green) and total protein stained with Sypro Ruby (shown in red). Images were artificially colored using PD Quest (BioRad) software tools. Phosphoprotein staining was confirmed with the visualization of the two positive molecular weight control bands (Peppermint Stick – Molecular Probes). The molecular weight (MW) of marker proteins and the pH range of the IEF gradient are indicated. N=3.</p

Proteins that presented the major phosphorylation changes in response to TGF-β.

<p>Magnified regions showing protein spots that presented the greatest up- (A) or down-(C) phosphoregulation and their respective bar graphics (B and D) with the mean fold change values.</p