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The COVID-19 pandemic has imposed several challenges and strains at all levels of the educational system, especially as a consequence of lockdown and social distance measures. After a period of exclusive use of the online educational environment, educators have adapted to the new circumstances and, by a combination of different strategies, have fought to overcome the limitations and deficiencies of virtual learning. Student motivation, productivity, and creativity continue to be the main pedagogical issues that have to be reached with the new didactic tools developed during the pandemic. At the same time, this pandemic has shown the importance of the inclusion of microbiology as a core element of the educational curriculum and the opportunity to raise public awareness of the importance of microbes to everyday life.

Gasdermin D (GSDMD) and mixed lineage kinase domain-like protein (MLKL) are the pore-forming effectors of pyroptosis and necroptosis, respectively, with the capacity to disturb plasma membrane selective permeability and induce regulated cell death. The budding yeast Saccharomyces cerevisiae has long been used as a simple eukaryotic model for the study of proteins associated with human diseases by heterologous expression. In this work, we expressed in yeast both GSDMD and its N-terminal domain (GSDMD(NT)) to characterize their cellular effects and compare them to those of MLKL. GSDMD(NT) and MLKL inhibited yeast growth, formed cytoplasmic aggregates and fragmented mitochondria. Loss-of-function point mutants of GSDMD(NT) showed affinity for this organelle. Besides, GSDMD(NT) and MLKL caused an irreversible cell cycle arrest through TORC1 inhibition and disrupted endosomal and autophagic vesicular traffic. Our results provide a basis for a humanized yeast platform to study GSDMD and MLKL, a useful tool for structure–function assays and drug discovery.

Toll-like receptor (TLR) signaling is key to detect pathogens and initiating inflammation. Ligand recognition triggers the assembly of supramolecular organizing centers (SMOCs) consisting of large complexes composed of multiple subunits. Building such signaling hubs relies on Toll Interleukin-1 Receptor (TIR) and Death Domain (DD) protein-protein interaction domains. We have expressed TIR domain-containing components of the human myddosome (TIRAP and MyD88) and triffosome (TRAM and TRIF) SMOCs in Saccharomyces cerevisiae, as a platform for their study. Interactions between the TLR4 TIR domain, TIRAP, and MyD88 were recapitulated in yeast. Human TIRAP decorated the yeast plasma membrane (PM), except for the bud neck, whereas MyD88 was found at cytoplasmic spots, which were consistent with endoplasmic reticulum (ER)-mitochondria junctions, as evidenced by co-localization with Mmm1 and Mdm34, components of the ER and Mitochondria Encounter Structures (ERMES). The formation of MyD88-TIRAP foci at the yeast PM was reinforced by co-expression of a membrane-bound TLR4 TIR domain. Mutations in essential residues of their TIR domains aborted MyD88 recruitment by TIRAP, but their respective subcellular localizations were unaltered. TRAM and TRIF, however, did not co-localize in yeast. TRAM assembled long PM-bound filaments that were disrupted by co-expression of the TLR4 TIR domain. Our results evidence that the yeast model can be exploited to study the interactions and subcellular localization of human SMOC components in vivo.

ObjectiveCOVID-19 Living OVerview of Evidence (COVID-19 L·OVE) is a public repository and classification platform for COVID-19 articles. The repository contains over 430,000 articles as of 20 September 2021 and intends to provide a one-stop shop for COVID-19 evidence. Considering that systematic reviews conduct high-quality searches, this study assesses the comprehensiveness and currency of the repository against the total number of studies in a representative sample of COVID-19 systematic reviews.MethodsOur sample was generated from all the studies included in the systematic reviews of COVID-19 published during April 2021. We estimated the comprehensiveness of COVID-19 L·OVE repository by determining how many of the individual studies in the sample were included in the COVID-19 L·OVE repository. We estimated the currency as the percentage of studies that were available in the COVID-19 L·OVE repository at the time the systematic reviews conducted their own search.ResultsWe identified 83 eligible systematic reviews that included 2132 studies. COVID-19 L·OVE had an overall comprehensiveness of 99.67% (2125/2132). The overall currency of the repository, that is, the proportion of articles that would have been obtained if the search of the reviews was conducted in COVID-19 L·OVE instead of searching the original sources, was 96.48% (2057/2132). Both the comprehensiveness and the currency were 100% for randomised trials (82/82).ConclusionThe COVID-19 L·OVE repository is highly comprehensive and current. Using this repository instead of traditional manual searches in multiple databases can save a great amount of work to people conducting systematic reviews and would improve the comprehensiveness and timeliness of evidence syntheses. This tool is particularly important for supporting living evidence synthesis processes

Toll-like receptor (TLR) signaling is key to detect pathogens and initiating inflammation. Ligand recognition triggers the assembly of supramolecular organizing centers (SMOCs) consisting of large complexes composed of multiple subunits. Building such signaling hubs relies on Toll Interleukin-1 Receptor (TIR) and Death Domain (DD) protein-protein interaction domains. We have expressed TIR domain-containing components of the human myddosome (TIRAP and MyD88) and triffosome (TRAM and TRIF) SMOCs in Saccharomyces cerevisiae, as a platform for their study. Interactions between the TLR4 TIR domain, TIRAP, and MyD88 were recapitulated in yeast. Human TIRAP decorated the yeast plasma membrane (PM), except for the bud neck, whereas MyD88 was found at cytoplasmic spots, which were consistent with endoplasmic reticulum (ER)-mitochondria junctions, as evidenced by co-localization with Mmm1 and Mdm34, components of the ER and Mitochondria Encounter Structures (ERMES). The formation of MyD88-TIRAP foci at the yeast PM was reinforced by co-expression of a membrane-bound TLR4 TIR domain. Mutations in essential residues of their TIR domains aborted MyD88 recruitment by TIRAP, but their respective subcellular localizations were unaltered. TRAM and TRIF, however, did not co-localize in yeast. TRAM assembled long PM-bound filaments that were disrupted by co-expression of the TLR4 TIR domain. Our results evidence that the yeast model can be exploited to study the interactions and subcellular localization of human SMOC components in vivo.

Caspases are a family of cysteine proteases that play an essential role in inflammation, apoptosis, cell death, and development. Here we delve into the effects caused by heterologous expression of human caspase-1 in the yeast Saccharomyces cerevisiae and compare them to those of caspase-8. Overexpression of both caspases in the heterologous model led to their activation and caused mitochondrial hyperpolarization, damage to different organelles, and cell death. All these effects were dependent on their protease activity, and caspase-8 was more aggressive than caspase-1. Growth arrest could be at least partially explained by dysfunction of the actin cytoskeleton as a consequence of the processing of the yeast Bni1 formin, which we identify here as a likely direct substrate of both caspases. Through the modulation of the GAL1 promoter by using different galactose:glucose ratios in the culture medium, we have established a scenario in which caspase-1 is sufficiently expressed to become activated while yeast growth is not impaired. Finally, we used the yeast model to explore the role of death-fold domains (DD) of both caspases in their activity. Peculiarly, the DDs of either caspase showed an opposite involvement in its intrinsic activity, as the deletion of the caspase activation and recruitment domain (CARD) of caspase-1 enhanced its activity, whereas the deletion of the death effector domain (DED) of caspase-8 diminished it. We show that caspase-1 is able to efficiently process its target gasdermin D (GSDMD) when co-expressed in yeast. In sum, we propose that S. cerevisiae provides a manageable tool to explore caspase-1 activity and structure–function relationships.

The yeast Saccharomyces cerevisiae is a model organism that has been thoroughly exploited to understand the universal mechanisms that govern signaling pathways. Due to its ease of manipulation, humanized yeast models that successfully reproduce the function of human genes permit the development of highly efficient genetic approaches for molecular studies. Of special interest are those pathways related to human disease that are conserved from yeast to mammals. However, it is also possible to engineer yeast cells to implement functions that are naturally absent in fungi. Along the years, we have reconstructed several aspects of the mammalian phosphatidylinositol 3-kinase (PI3K) pathway in S. cerevisiae. Here, we briefly review the use of S. cerevisiae as a tool to study human oncogenes and tumor suppressors, and we present an overview of the models applied to the study of the PI3K oncoproteins, the tumor suppressor PTEN, and the Akt protein kinase. We discuss the application of these models to study the basic functional properties of these signaling proteins, the functional assessment of their clinically relevant variants, and the design of feasible platforms for drug discovery.

Caspases are a family of cysteine proteases that play an essential role in inflammation, apoptosis, cell death, and development. Here we delve into the effects caused by heterologous expression of human Caspase-1 in the yeast Saccharomyces cerevisiae and compare them to those of Caspase-8. Overexpression of both caspases in the heterologous model led to their activation, and caused mitochondrial depolarization, ROS production, damage to different organelles, and cell death. All these effects were dependent on their protease activity, and Caspase-8 was more aggressive than Caspase-1. Growth arrest could be at least partially explained by dysfunction of the actin cytoskeleton as a consequence of the processing of the yeast Bni1 formin, which we identify here as a likely direct substrate of both caspases. Through the modulation of the GAL1 promoter by using different galactose:glucose ratios in the culture medium, we have established a scenario in which Caspase-1 is sufficiently expressed to become activated while yeast growth is not impaired. Finally, we used the yeast model to explore the role of death-fold domains (DD) of both caspases in their activity. Peculiarly, the DDs of either caspase showed an opposite involvement in its intrinsic activity, as the deletion of the caspase activation and recruitment domain (CARD) of Caspase-1 enhanced its activity, while the deletion of the death effector domain (DED) of Caspase-8 diminished it. We propose the yeast model as a useful and manageable tool to explore Caspase-1 structure-function relationships, the impact of mutations or the activity of putative inhibitors or regulators.

The PTEN tumor suppressor gene is mutated with high incidence in tumors and in the germline of patients with cancer predisposition or with macrocephaly associated to autism. PTEN nonsense mutations generating premature termination codons (PTC) and producing non-functional truncated PTEN proteins are frequent in association with human disease. However, there are no studies addressing the restoring of full-length PTEN proteins from PTC-mutated PTEN gene by translational readthrough. Here, we have performed a global translational and functional readthrough analysis of the complete collection of PTEN PTC somatic or hereditary mutations found in tumors or in the germline of patients (disease-associated PTEN PTCome), and we set standards for the analysis of the potential of readthrough functional reconstitution in disease-relevant genes. Our analysis indicates that prevalent pathogenic PTEN PTC mutations are susceptible of PTEN functional restoration in response to readthrough-inducing compounds. Comprehensive readthrough analyses of disease-associated PTComes will be valuable tools for implementation of readthrough-based precision interventions in specific groups of patients. This article is protected by copyright. All rights reserved.

The pipeline transport system (PTS) is a means of land and marine transport, reliable in the oil sector as well as in various industries, considering its economy and speed to transport a product. The instability of this system, though, currently takes several variables, which directly affect the mechanical integrity (MI) of the same, due to the imminent internal and external formation of the phenomenon classified as corrosion (C). To address this problem, fault tolerance control techniques (FTCS), global positioning systems (GPS), and artificial intelligence (AI), are useful tools to deal with uncertainty and minimize subjectivity, through system modeling. That will allow the identification, control, and timely monitoring of risk factors. During the last 30 years, important contributions on corrosion to pipelines in the oil and gas industry have been published. This document aims to build a systematic literary review (SLR), from an integrative perspective, recognizing various studies in the periods 1987–2018, analyzing 245 documents, from three posts previously not addressed together within the context of corrosion to PTS, offering an apology if any document relevant to the subject was excluded from the work. Findings about the topics of analysis, discussions, and future lines of research are described.

Brucella species are facultative intracellular Gram-negative bacteria relevant to animal and human health. Their ability to establish an intracellular niche and subvert host cell pathways to their advantage depends on the delivery of bacterial effector proteins through a type IV secretion system. Brucella Toll/Interleukin-1 Receptor (TIR)-domain-containing proteins BtpA (also known as TcpB) and BtpB are among such effectors. Although divergent in primary sequence, they interfere with Toll-like receptor (TLR) signaling to inhibit the innate immune responses. However, the molecular mechanisms implicated still remain unclear. To gain insight into the functions of BtpA and BtpB, we expressed them in the budding yeast Saccharomyces cerevisiae as a eukaryotic cell model. We found that both effectors were cytotoxic and that their respective TIR domains were necessary and sufficient for yeast growth inhibition. Growth arrest was concomitant with actin depolymerization, endocytic block and a general decrease in kinase activity in the cell, suggesting a failure in energetic metabolism. Indeed, levels of ATP and NAD+ were low in yeast cells expressing BtpA and BtpB TIR domains, consistent with the recently described enzymatic activity of some TIR domains as NAD+ hydrolases. In human epithelial cells, both BtpA and BtpB expression reduced intracellular total NAD levels. In infected cells, both BtpA and BtpB contributed to reduction of total NAD, indicating that their NAD+ hydrolase functions are active intracellularly during infection. Overall, combining the yeast model together with mammalian cells and infection studies our results show that BtpA and BtpB modulate energy metabolism in host cells through NAD+ hydrolysis, assigning a novel role for these TIR domain-containing effectors in Brucella pathogenesis., Author summary Brucella is a genus of zoonotic bacteria that cause severe disease in a variety of mammals, ranging from farm animals (as bovines, swine and ovine) to marine mammals. Transmission to humans, often by ingestion of non-treated dairy products, leads to serious systemic infection. Brucella abortus invades host cells and replicates intracellularly. Such behavior relies on the injection of bacterial proteins into the host cytoplasm via specialized secretion systems. Our work focuses on the study of two of these factors, BtpA and BtpB, previously described to contain Toll/Interleukin-1 Receptor (TIR)-domains that modulate innate immunity. We use here two biological models: the yeast Saccharomyces cerevisiae and human cell lines. We found that the TIR domains of both Brucella proteins were necessary and sufficient to collapse energy metabolism in yeast by depleting ATP and NAD+. This result was translatable to higher cells and consistent with the recently described NADase activity of some TIR domains both in mammalian and bacterial proteins. Importantly, we demonstrate that Brucella down-regulates total NAD levels in host cells by using both BtpA and BtpB effectors. Our results show that NAD+ is targeted by Brucella during infection, which may constitute a novel mechanism for its pathogenicity.

Small World Initiative and Tiny Earth are popular citizen science programs that are implemented worldwide in response to the global antibiotic resistance crisis. When starting up the program in Albacete (Spain), we noted that rates of isolated antibiotic-producing bacteria are generally low. To make the activity more stimulating for participating students, we modified the protocol to obtain more positive results by focusing on isolation of actinomycetes, the main producers of most clinically used antibiotics. Adaptations involved redesigning culture media, incubation times and temperatures, and modification of the ESKAPE antibiosis experiment by employing an agar-transplantation step. Of 390 bacterial isolates tested, almost 6% tested positive in antibiosis experiments and DNA sequence analysis confirmed that all positives are actinomycetes, demonstrating that our protocol is efficient toward isolating antibiotic-producing actinomycetes from soil. Evaluation forms filled by participating students indicated that the program was received very positively and that our modifications contribute to make this educational program more stimulating and efficient.

espanolLa produccion bibliografica del siglo XVIII se sirvio de las Gacetas de Mexico para la promocion de autores y titulos de impresos que aparecian cotidianamente. Bajo los titulos Libros nuevos de Mexico, Libros nuevos de Espana o Libros nuevos, varios de los fasciculos de las Gacetas ofrecen a los lectores informacion sobre novedades bibliograficas. El presente texto se enfoca particularmente en la identificacion del conjunto de noticias sobre impresos espanoles que aparecen en las dos primeras Gacetas de Mexico, que en total representan 15 anos de publicacion y mas de 160 fasciculos. Se ofrecen datos sobre autores, titulos, temas y lugares de venta de los libros espanoles que circularon en Mexico durante la primera mitad del siglo XVIII. EnglishBibliographic production in the 18th century utilized the Gacetas de Mexico to promote the authors and printed titles that appearing daily under such headings as “New Mexican Books”, “New Spanish Books”, or “New Books” (Libros nuevos de Mexico, Libros nuevos de Espana, Libros nuevos). Several fascicles of the Gacetas offered to their readers information on recent publications. This paper focuses particularly on the identification of all the news on Spanish print appearing in the first two Gazettes of Mexico, which together represent 15 years of publication and over 160 fascicles. Data on authors, titles, subjects and places selling Spanish books that circulated in Mexico during the first half of the eighteenth century.

Hematopoietic stem cell transplantation (HSCT) from a mismatched unrelated donor, an haploidentical donor or a cord blood unit (CBU) has become a widely aviable approach if patient lacks a matched related or unrelated donor. However, if the patient has anti-HLA antibodies against antigens present in the mismatched donor or CBU (donor-specific antibodies, DSAs) this option should be disregarded due to the high risk of graft failure. Desensitization can be used to reduce levels of DSAs but this technique has limited results. We report the case of a 62-year-old woman with DSAs against two haploidentical familiar donors who failed desensitization of DSAs. Finally she underwent a HSCT from a 5/10 mismatched unrelated donor which has been successful.

Las reacciones donde intervienen oxidantes y radicales han jugado un papel esencial en el origen de las formas de vida aerobias y son una parte integral de la homeostasis celular. Sin embargo y debido a los efectos tóxicos colaterales de esta presión oxidativa, de forma muy temprana en la evolución se desarrollaron los enzimas y factores antioxidantes que son capaces de controlar la presencia y efectos de estos productos. Oxidantes y antioxidantes tienen una clara función en el organismo y un desequilibrio en este delicado balance resulta en muchas alteraciones bioquímicas y celulares que pueden crear condiciones patológicas. Así mismo es creciente el papel que se le da a los radicales libres del oxígeno como factor de envejecimiento biológico (1). Es difícil encontrar actualmente algún campo de la biomedicina donde no pueda encontrarse trabajos relacionados con oxidantes y radicales libres. En los organismos vivos los principales oxidantes y radicales libres son los derivados del oxígeno, que genéricamente denominaremos intermediarios reactivos del oxígeno (IRO). Si bien tenemos una idea acerca de estos compuestos, sus amplias repercusiones y recientes implicaciones en distintas patologías, así como la progresiva complejidad de sus interacciones y efectos celulares justifican, a nuestro juicio, una revisión que intente dar una visión de conjunto sobre estos inevitables, útiles y peligrosos reactivos.

Background Social cognition and metacognition have emerged as cornerstones of research and treatment of schizophrenia. Both constructs are known to influence the onset of psychosis, to predict functional outcome and to be associated with symptoms. However, whether the deficits in first-episode psychosis are homogeneous or group in patterns remains to be studied. This study aimed to analyze patterns of social cognitive and metacognitive variables in a sample of subjects with first-episode psychosis. Methods We recruited 192 subjects with first-episode psychosis from ten public mental-health services in Spain. We collected: demographic information, measures of functioning, performance in social cognition (the Faces Test, IPSAQ and the Hinting Task), a battery of metacognitive tasks (BCIS, and the Beads Task) and a neuropsychological assessment. We performed a Latent Profile Analysis (LPA) with the metacognitive and social-cognitive variables. The variable importance was assessed via a classification tree (CART) and the mean differences among the resulting groups for clinical, neuropsychological and functioning variables were calculated with ANOVA and Kruskal-Wallis tests. Results Our sample was comprised of 192 (62 women) with first-episode psychosis. The mean age of the sample was 27.93(1.39). The mean PANSS total score of the sample was 58.48 (17.79). The average GAF score was 58.93 (12.25). We included 174 cases with complete social-cognitive and metacognitive data in the cluster analysis. We identified three type-VEE clusters (i.e. ellipsoidal clusters with equal shape and orientation) according to BIC (BIC=-3600.651). The 85-15 condition of the Beads Task and the Hinting Task emerged as the most important variables in determining the clustering structure. The first cluster (60.9%) was characterized by average scores in most of the metacognitive and social cognitive variables, but the presence of the jumping to conclusions bias. The second cluster (5.7%) was characterized by low self-reflectiveness, presence of personalizing bias and an excessive number of trials in the beads task. The third cluster (33.5%) was characterized by average scores in all metacognitive tasks but low scores in the social cognitive tasks. Discussion We found three clusters in a large sample of subjects with first-episode psychosis. Our results indicate that the three groups differ in the proneness to present deficits in specific domains. Furthermore, in our sample, patients may not exhibit a homogeneous deficit in all social-cognitive and metacognitive variables. Instead, the impairment may be particularly prominent in either social-cognitive or metacognitive variables. Subjects in different clusters may present differences in their clinical characteristics, what could be relevant in the treatment. Therefore, with further research, a thorough assessment of social cognition and metacognition may help personalize the treatment according to the person’s subtype of the deficit.

Brucellaspecies are facultative intracellular Gram-negative bacteria relevant to animal and human health. Their ability to establish an intracellular niche and subvert host cell pathways to their advantage depends on the delivery of bacterial effector proteins through a type IV secretion system.BrucellaToll/Interleukin-1 Receptor (TIR)-domain-containing proteins BtpA (also known as TcpB) and BtpB are among such effectors. Although divergent in primary sequence, they interfere with Toll-like receptor (TLR) signaling to inhibit the innate immune responses. However, the molecular mechanisms implicated still remain unclear. To gain insight into the functions of BtpA and BtpB, we expressed them in the budding yeastSaccharomyces cerevisiaeas a eukaryotic cell model. We found that both effectors were cytotoxic and that their respective TIR domains were necessary and sufficient for yeast growth inhibition. Growth arrest was concomitant with actin depolymerization, endocytic block and a general decrease in kinase activity in the cell, suggesting a failure in energetic metabolism. Indeed, levels of ATP and NAD+were low in yeast cells expressing BtpA and BtpB TIR domains, consistent with the recently described enzymatic activity of some TIR domains as NAD+hydrolases. In human epithelial cells, both BtpA and BtpB expression reduced intracellular total NAD levels. In infected cells, both BtpA and BtpB contributed to reduction of total NAD, indicating that their NAD+hydrolase functions are active intracellularly during infection. Overall, combining the yeast model together with mammalian cells and infection studies our results show that BtpA and BtpB modulate energy metabolism in host cells through NAD+hydrolysis, assigning a novel role for these TIR domain-containing effectors inBrucellapathogenesis.

Klebsiella pneumoniae is recognized as an urgent threat to human health due to the increasing isolation of multidrug resistant strains. Hypervirulent strains are a major concern due to their ability to cause life-threating infections in healthy hosts. The type VI secretion system (T6SS) is widely implicated in microbial antagonism, and it mediates interactions with host eukaryotic cells in some cases. In silico search for genes orthologous to T6SS component genes and T6SS effector genes across 700 K. pneumoniae genomes shows extensive diversity in T6SS genes across the K. pneumoniae species. Temperature, oxygen tension, pH, osmolarity, iron levels, and NaCl regulate the expression of the T6SS encoded by a hypervirulent K. pneumoniae strain. Polymyxins and human defensin 3 also increase the activity of the T6SS. A screen for regulators governing T6SS uncover the correlation between the transcription of the T6SS and the ability to kill E. coli prey. Whereas H-NS represses the T6SS, PhoPQ, PmrAB, Hfq, Fur, RpoS and RpoN positively regulate the T6SS. K. pneumoniae T6SS mediates intra and inter species bacterial competition. This antagonism is only evident when the prey possesses an active T6SS. The PhoPQ two component system governs the activation of K. pneumoniae T6SS in bacterial competitions. Mechanistically, PhoQ periplasmic domain, and the acid patch within, is essential to activate K. pneumoniae T6SS. Klebsiella T6SS also mediates anti-fungal competition. We have delineated the contribution of each of the individual VgrGs in microbial competition and identified VgrG4 as a T6SS effector. The DUF2345 domain of VgrG4 is sufficient to intoxicate bacteria and yeast. ROS generation mediates the antibacterial effects of VgrG4, and the antitoxin Sel1E protects against the toxic activity of VgrG4. Our findings provide a better understanding of the regulation of the T6SS in bacterial competitions, and place ROS as an early event in microbial competition., Author summary Klebsiella pneumoniae has been singled out as an “urgent threat to human health” due to extremely drug resistant strains. Numerous studies investigate the molecular mechanisms underlying antibiotic resistance in K. pneumoniae, while others dissect the virulence strategies of this pathogen. However, there is still limited knowledge on the fitness of Klebsiella in the environment, and, particularly, the competition of Klebsiella with other species. Here, we demonstrate that Klebsiella exploits the type VI secretion system (T6SS) nanoweapon to kill bacterial competitors and fungi. K. pneumoniae perceives T6SS attacks from bacterial competitors, resulting in retaliation against the aggressive cell. The perception of the attack involved the sensor PhoPQ and led to the up-regulation of the T6SS. We identified one of the toxins deployed by the T6SS to antagonize other microbes and revealed how Klebsiella protects itself from this toxin. Our findings provide a better understanding of the T6SS role in microbial competition and uncover new aspects on how bacteria regulate T6SS-mediated microbial antagonism.

It remains to be seen how the recent October 2018 election will affect Brazil’s economy and foreign multinational corporations aiming to enter and operate in this market. The election has offered Brazilian voters two extreme political choices: Fernando Haddad of the left-wing Partido dos Trabalhadores (PT) party, supported by the popular former president Luiz Inacio Lula da Silva who campaigned for Haddad out of prison while serving his 12-year sentence, and Jair Bolsonaro of the Partido Social Liberal (PSL), a right-wing populist. The election of the latter as the new president, which resulted in a drastic shift of Brazilian politics to the right, raised both concerns and hope in the country. President Bolsonaro’s populist campaign, which has widely been compared with the populist campaign and political rhetoric of US President Donald J. Trump, and the selection of a far-right former army captain as Vice President, one who shares Bolsonaro’s nostalgia for Brazil’s military dictatorship from 1964–1985, are reasons for considerable concerns about a new wave of far-right, authoritarian-style government in the country as well as in Latin America (a wave that has also recently been evident in other parts of the globe). However, the country’s moribund economy and systemically corrupt political environment called for a change in Brazil after nearly 15 years of left-wing rule. The new president’s selection of a University of Chicago-trained free-market economist, Paulo Guedes, as his economic advisor and new finance minister is sending signals of hope that the long-needed economic and financial reforms will be realized, similar to the Milton Friedman-style free-market reforms that were implemented in Chile under General Augusto Pinochet, who came to power in 1973 after overthrowing socialist president Salvador Allende.

Sclerodermoid chronic graft-versus-host disease (scGVHD) is a severe complication of allogeneic haema­- topoietic stem cell transplantation. The aim of this study was to investigate the usefulness of high-frequency ultrasound of the skin in assessing the inflammatory patterns and prognosis of patients with scGVHD. A prospective study was carried out with patients who developed scGVHD in the period June 2016 to April 2018. Clinical and ultrasound examinations were performed on the first visit and at 6-month follow-up. A total of 24 patients were included in the study. A 6-month follow-up high-frequency ultrasound of the skin was performed on 20 of the 24 patients. Abnormal B-mode findings in high-frequency ultrasound of the skin consisted of hypoechogenic dermis, hypoechogenicity of septa and hyperechogenicity of lobules in hypodermis. No differences were observed in these basal parameters between treatment progressive/non-responding and inactive/responding scGVHD groups of patients. Basal Doppler showing increased vascular flow with a systolic peak ≥10 cm/s and a vascular resistance index ≥ 0.70 was observed only in those patients who developed progressive/non-responding scGVHD (62.5% vs 0% p = 0.006). In conclusion, Doppler ultrasound is a useful tool to assess the inflammatory activity and outcome of scGVHD. These findings could enhance patient management and help to guide treatment decisions.

Many bacterial pathogens use specialized secretion systems to deliver virulence effector proteins into eukaryotic host cells. The function of these effectors depends on their localization within infected cells, but the mechanisms determining subcellular targeting of each effector are mostly elusive. Here, we show that the Salmonella type III secretion effector SteA binds specifically to phosphatidylinositol 4-phosphate [PI(4)P]. Ectopically expressed SteA localized at the plasma membrane (PM) of eukaryotic cells. However, SteA was displaced from the PM of Saccharomyces cerevisiae in mutants unable to synthesize the local pool of PI(4)P and from the PM of HeLa cells after localized depletion of PI(4)P. Moreover, in infected cells, bacterially translocated or ectopically expressed SteA localized at the membrane of the Salmonella-containing vacuole (SCV) and to Salmonella-induced tubules; using the PI(4)P-binding domain of the Legionella type IV secretion effector SidC as probe, we found PI(4)P at the SCV membrane and associated tubules throughout Salmonella infection of HeLa cells. Both binding of SteA to PI(4)P and the subcellular localization of ectopically expressed or bacterially translocated SteA were dependent on a lysine residue near the N-terminus of the protein. Overall, this indicates that binding of SteA to PI(4)P is necessary for its localization within host cells.

Platelet rich plasma (PRP) method was the first method used for platelet concentrate (PC) preparation and continues to be the only licensed method in the USA. By contrast in Europe since the eighties of the last century, the PRP method has been progressively replaced by methods based on the removal of the buffy coat (BC) layer. Initially BC were processed individually to produced a single PC but it was soon recognized that the pooling of 4–6 BC units with the addition of a plasma unit of one of the donors of the pool or a platelet additive solution (PAS), improved the efficiency of the separation and eased the transfusion process since the final product was stored as an adult transfusion dose ready to be connected to the patient. Semi-automatic devices were developed for the separation of whole blood into red blood cells, plasma and the BC and for the separation of the BC into the PC. The introduction of PAS in combination with plasma (in general about 35% of plasma and 65% PAS) for storing PC has shown to offer advantages such as an increased availability of plasma for fractionation, a reduction of transfusion reactions to platelets and a potential reduction in the risk of transfusion related acute lung injury. While early generations of PAS were associated to a decrease in the post-transfusion corrected platelet count in comparison to PC stored in 100% plasma, this effect has not been found with the newer generation of PAS.

Although platelet transfusions continue to be the mainstay in the management of patients suffering from qualitative and quantitative disorders, several strategies might be considered when trying to reduce or avoid platelet transfusions. Those include assuring a higher haemoglobin level and pharmacological interventions. Several studies in vitro and in vivo suggest that higher haematocrit decreases bleeding time in anaemic patients. This effect might be due to the haemorheological effect of red blood cells that expel platelets towards the wall optimizing the interaction of circulating platelets with the injured subendothelium. Several drugs have shown the capacity for improving the haemostasis and decreasing the bleeding diathesis in patients suffering from thrombocytopathies and/or thrombocytopenias, although the evidence is scarce and sometimes contradictory. Among antifibrinolytic drugs, two synthetic derivates are available, epsilon aminocaproic acid and tranexamic acid, although the latter one is the preferred for most of indications, given its higher and more sustained antifibrinolytic activity in tissues and lower and less frequent dosing. A few reports have shown the efficacy of desmopressin in treating patients with quantitative platelet disorders and Bernard–Soulier syndrome. Patients with Glanzmann thrombasthenia do not respond to desmopressin. Recombinant activated factor VII is currently approved in the European Union for treating patients with Glanzmann thrombasthenia refractory to platelet transfusions due to antibodies to GPIIb-IIIa and/or HLA. Currently, we have available two thrombopoietin analogues (romiplostim and eltrombopag) that have been used in patients with quantitative and/or qualitative platelet disorders. Several platelet substitutes are currently under development and one is being tested in healthy volunteers, but none is available yet in the market.

The Small World Initiative (SWI) and Tiny Earth are a consolidated and successful education programs rooted in the USA that tackle the antibiotic crisis by a crowdsourcing strategy. Based on active learning, it challenges young students to discover novel bioactive-producing microorganisms from environmental soil samples. Besides its pedagogical efficiency to impart microbiology content in academic curricula, SWI promotes vocations in research and development in Experimental Sciences and, at the same time, disseminates the antibiotic awareness guidelines of the World Health Organization. We have adapted the SWI program to the Spanish academic environment by a pioneering hierarchic strategy based on service-learning that involves two education levels (higher education and high school) with different degrees of responsibility. Throughout the academic year, 23 SWI teams, each consisting of 3-7 undergraduate students led by one faculty member, coordinated off-campus programs in 22 local high schools, involving 597 high school students as researchers. Post-survey-based evaluation of the program reveals a satisfactory achievement of goals: acquiring scientific abilities and general or personal competences by university students, as well as promoting academic decisions to inspire vocations for science- and technology-oriented degrees in younger students, and successfully communicating scientific culture in antimicrobial resistance to a young stratum of society.

The Akt protein kinase is the main transducer of phosphatidylinositol-3,4,5-trisphosphate (PtdIns3,4,5P3) signaling in higher eukaryotes, controlling cell growth, motility, proliferation and survival. By co-expression of mammalian class I phosphatidylinositol 3-kinase (PI3K) and Akt in the Saccharomyces cerevisiae heterologous model, we previously described an inhibitory effect on yeast growth that relied on Akt kinase activity. Here we report that PI3K-Akt expression in yeast triggers the formation of large plasma membrane (PM) invaginations that were marked by actin patches, enriched in PtdIns4,5P2 and associated to abnormal intracellular cell wall deposits. These effects of Akt were mimicked by overproduction of the PtdIns4,5P2 effector Slm1, an adaptor of the Ypk1 and Ypk2 kinases in the TORC2 pathway. Although Slm1 was phosphorylated in vivo by Akt, TORC2-dependent Ypk1 activation did not occur. However, PI3K-activated Akt suppressed the lethality derived from inactivation of either TORC2 or Ypk protein kinases. Thus, heterologous co-expression of PI3K and Akt in yeast short-circuits PtdIns4,5P2- and TORC2-signaling at the level of the Slm-Ypk complex, overriding some of its functions. Our results underscore the importance of phosphoinositide-dependent kinases as key actors in the homeostasis and dynamics of the PM.

Background Automation of blood component preparation (BCP) from whole blood (WB) collections can help to optimize the BCP process, and it is increasingly being widespread implemented. Aim This review summarizes the quality of blood components obtained with new automated devices developed in the 2000s. Methods We reviewed available literature on the quality of blood components obtained with new automated devices. Results Blood components obtained with the new devices met European standards. Of note, compared with platelet concentrates obtained with manual methods, automation of BCP improved the consistency of the final products. Conclusion The complete automation of BCP from WB collections is still in development, and it represents a huge change in paradigm.

The PTEN phosphoinositide 3-phosphatase is a tumor suppressor commonly targeted by pathologic missense mutations. Subject to multiple complex layers of regulation, its capital role in cancer relies on its counteracting function of class I phosphoinositide 3-kinase (PI3K), a key feature in oncogenic signaling pathways. Precise assessment of the involvement of PTEN mutations described in the clinics in loss of catalytic activity requires either tedious in vitro phosphatase assays or in vivo experiments involving transfection into mammalian cell lines. Taking advantage of the versatility of the model organism Saccharomyces cerevisiae, we have developed different functional assays by reconstitution of the mammalian PI3K-PTEN switch in this lower eukaryote. This methodology is based on the fact that regulated PI3K expression in yeast cells causes conversion of PtdIns(4,5)P2 in PtdIns(3,4,5)P3 and co-expression of PTEN counteracts this effect. This can be traced by monitoring growth, given that PtdIns(4,5)P2 pools are essential for the yeast cell, or by using fluorescent reporters amenable for microscopy or flow cytometry. Here we describe the methodology and review its application to evaluate the functionality of PTEN mutations. We show that the technique is amenable to both directed and systematic structure-function relationship studies, and present an example of its use for the study of the recently discovered PTEN-L variant.

The PTEN gene encodes a master regulator protein that exerts essential functions both in the cytoplasm and in the nucleus. PTEN is mutated in the germline of both patients with heterogeneous tumor syndromic diseases, categorized as PTEN hamartoma tumor syndrome (PHTS), and a group affected with autism spectrum disorders (ASD). Previous studies have unveiled the functional heterogeneity of PTEN variants found in both patient cohorts, making functional studies necessary to provide mechanistic insights related to their pathogenicity. Here, we have functionally characterized a PTEN missense variant [c.49C>G; p.(Gln17Glu); Q17E] associated to both PHTS and ASD patients. The PTEN Q17E variant displayed partially reduced PIP3-catalytic activity and normal stability in cells, as shown using S. cerevisiae and mammalian cell experimental models. Remarkably, PTEN Q17E accumulated in the nucleus, in a process involving the PTEN N-terminal nuclear localization sequence. The analysis of additional germline-associated PTEN N-terminal variants illustrated the existence of a PTEN N-terminal region whose targeting in disease causes PTEN nuclear accumulation, in parallel with defects in PIP3-catalytic activity in cells. Our findings highlight the frequent occurrence of PTEN gene mutations targeting PTEN N-terminus whose pathogenicity may be related, at least in part, with the retention of PTEN in the nucleus. This could be important for the implementation of precision therapies for patients with alterations in the PTEN pathway.

López-Goñi, Ignacio et al., Social networks are already being exploited for searching, storing, and sharing knowledge, demonstrating that they are an efficient vehicle for social learning. Consequently, they could be implemented as a competent tool for formal learning. Twitter is among the 10 most popular online social networks, integrating a community of over 500 million users around the world. Twitter has already been used in several educational programs and evaluated as a positive teaching experience with an outstanding potential in academic and educational environments (1–6). However, there are scarce examples available in the literature about its use in science teaching and communication. In this work, we present and analyze the application of Twitter to create an online space for communication and learning of basic microbiology. With this aim, a team of professionals in the field has imparted, to our knowledge, the first worldwide open access microbiology course via Twitter. Here we assess the results of our experience of using this social network as a tool for teaching, promoting, and communicating scientific knowledge to a wide audience.

In higher eukaryotes, cell proliferation is regulated by class I phosphatidylinositol 3-kinase (PI3K), which transduces stimuli received from neighboring receptors by local generation of PtdIns(3,4,5)P3 in cellular membranes. PI3K is a heterodimeric protein consisting of a regulatory and a catalytic subunit (p85 and p110 respectively). Heterologous expression of p110α in Saccharomyces cerevisiae leads to toxicity by conversion of essential PtdIns(4,5)P2 into futile PtdIns(3,4,5)P3, providing a humanized yeast model for functional studies on this pathway. Here, we report expression and functional characterization in yeast of all regulatory and catalytic human PI3K isoforms, and exploitation of the most suitable setting to functionally assay panels of tumor- and germ line-associated PI3K mutations, with indications to the limits of the system. The activity of p110α in yeast was not compromised by truncation of its N-terminal adaptor-binding domain (ABD) or inactivation of the Ras-binding domain (RBD). In contrast, a cluster of positively charged residues at the C2 domain was essential. Expression of a membrane-driven p65α oncogenic-truncated version of p85α, but not the full-length protein, led to enhanced activity of α, β, and δ p110 isoforms. Mutations impairing the inhibitory regulation exerted by the p85α iSH2 domain on the C2 domain of p110α yielded the latter non-responsive to negative regulation, thus reproducing this oncogenic mechanism in yeast. However, p85α germ line mutations associated with short stature, hyperextensibility of joints and/or inguinal hernia, ocular depression, Rieger anomaly, and teething delay (SHORT) syndrome did not increase PI3K activity in this model, supporting the idea that SHORT syndrome-associated p85α mutations operate through mechanisms different from the canonical disruption of inhibitory p85–p110 interactions typical of cancer.

In recent years the imagen processing is an envolving issue, allows a great variety of applications. This article describe the process of automation of a conveyor belt using a machine learning algorithms that is able to immediately recognize each type different electronic board. For image processing a webcam is used. The obtained information is sent a free server of a web page by a WiFi module. Is obtained an automatic and intelligent process of low cost of robotic cell.