Identification of a fibrinogen-related protein (FBN9) gene in neotropical anopheline mosquitoes

<p>Abstract</p> <p>Background</p> <p>Malaria has a devastating impact on worldwide public health in many tropical areas. Studies on vector immunity are important for the overall understanding of the parasite-vector interaction and for the design of novel strategies to control malaria. A member of the fibrinogen-related protein family, <it>fbn9</it>, has been well studied in <it>Anopheles gambiae </it>and has been shown to be an important component of the mosquito immune system. However, little is known about this gene in neotropical anopheline species.</p> <p>Methods</p> <p>This article describes the identification and characterization of the <it>fbn9 </it>gene partial sequences from four species of neotropical anopheline primary and secondary vectors: <it>Anopheles darlingi, Anopheles nuneztovari, Anopheles aquasalis</it>, and <it>Anopheles albitarsis </it>(namely <it>Anopheles marajoara</it>). Degenerate primers were designed based on comparative analysis of publicly available <it>Aedes aegypti </it>and <it>An. gambiae </it>gene sequences and used to clone putative homologs in the neotropical species. Sequence comparisons and Bayesian phylogenetic analyses were then performed to better understand the molecular diversity of this gene in evolutionary distant anopheline species, belonging to different subgenera.</p> <p>Results</p> <p>Comparisons of the <it>fbn9 </it>gene sequences of the neotropical anophelines and their homologs in the <it>An. gambiae </it>complex (Gambiae complex) showed high conservation at the nucleotide and amino acid levels, although some sites show significant differentiation (non-synonymous substitutions). Furthermore, phylogenetic analysis of <it>fbn9 </it>nucleotide sequences showed that neotropical anophelines and African mosquitoes form two well-supported clades, mirroring their separation into two different subgenera.</p> <p>Conclusions</p> <p>The present work adds new insights into the conserved role of <it>fbn9 </it>in insect immunity in a broader range of anopheline species and reinforces the possibility of manipulating mosquito immunity to design novel pathogen control strategies.</p

School Evasion in the Brazilian trends: analyzing the vectors that influence students’ decision to interrupt their formative process

Research data demonstrate that the analyzes built around School Evasion considers, primarily, the quantitative metrics of studentsrsquo; entry and exit and also the monetary losses, especially because it would be incoherent not to take as reference the goals and objectives outlined for education school system. However, analyzing the vectors that influence studentsrsquo; decision to interrupt their formative process, according to purely numerical criteria, would imply in ignoring the function of educational institutions and the real causes/reasons stemming from the social and relational demand of students' passage on educational institutions. For this reason, problematizing some of the recurring issues and incidents generated by certain investigations seems instigating and challenging. If in on the one hand, the attitude of understanding the conceptualization, the investigative tendencies and the characteristics of the studies give rise to certain criticisms that put in check the complexity of the object in question, on the other, it can stimulate the construction of other tendencies, new pathways, other possible ways of overcoming the gaps identified in the studies about School Evasion in Brazil

The Impact of the Serum Extraction Protocol on Metabolomic Profiling Using UPLC-MS/MS and FTIR Spectroscopy

Funding Information: This research was funded by Fundação para a Ciência e a Tecnologia (FCT), Grants DSAIPA/DS/0117/2020 and RNEM-LISBOA-01-0145-FEDER-022125 (Portuguese Mass Spectrometry Network). The Centro de Química Estrutural is a Research Unit funded by FCT through projects UIDB/00100/2020 and UIDP/00100/2020. The Institute of Molecular Sciences is an Associate Laboratory funded by FCT through project LA/P/0056/2020. Publisher Copyright: © 2023 The Authors. Published by American Chemical Society.Biofluid metabolomics is a very appealing tool to increase the knowledge associated with pathophysiological mechanisms leading to better and new therapies and biomarkers for disease diagnosis and prognosis. However, due to the complex process of metabolome analysis, including the metabolome isolation method and the platform used to analyze it, there are diverse factors that affect metabolomics output. In the present work, the impact of two protocols to extract the serum metabolome, one using methanol and another using a mixture of methanol, acetonitrile, and water, was evaluated. The metabolome was analyzed by ultraperformance liquid chromatography associated with tandem mass spectrometry (UPLC-MS/MS), based on reverse-phase and hydrophobic chromatographic separations, and Fourier transform infrared (FTIR) spectroscopy. The two extraction protocols of the metabolome were compared over the analytical platforms (UPLC-MS/MS and FTIR spectroscopy) concerning the number of features, the type of features, common features, and the reproducibility of extraction replicas and analytical replicas. The ability of the extraction protocols to predict the survivability of critically ill patients hospitalized at an intensive care unit was also evaluated. The FTIR spectroscopy platform was compared to the UPLC-MS/MS platform and, despite not identifying metabolites and consequently not contributing as much as UPLC-MS/MS in terms of information concerning metabolic information, it enabled the comparison of the two extraction protocols as well as the development of very good predictive models of patient’s survivability, such as the UPLC-MS/MS platform. Furthermore, FTIR spectroscopy is based on much simpler procedures and is rapid, economic, and applicable in the high-throughput mode, i.e., enabling the simultaneous analysis of hundreds of samples in the microliter range in a couple of hours. Therefore, FTIR spectroscopy represents a very interesting complementary technique not only to optimize processes as the metabolome isolation but also for obtaining biomarkers such as those for disease prognosis.publishersversionpublishe

Multi-biologic group analysis for an ecosystem response to longitudinal river regulation gradients

This work assesses the effects of river regulation on the diversity of different instream and riparian biological communities along a relieve gradient of disturbance in regulated rivers. Two case studies in Portugal were used, with different river regulation typology (downstream of run-of-river and reservoir dams), where regulated and free-flowing river stretches were surveyed for riparian vegetation, macrophytes, bryophytes, macroalgae, diatoms and macroinvertebrates. The assessment of the regulation effects on biological communities was approached by both biological and functional diversity analysis. Results of this investigation endorse river regulation as a major factor differentiating fluvial biological communities through an artificial environmental filtering that governs species assemblages by accentuating species traits related to river regulation tolerance. Communities' response to regulation gradient seem to be similar and insensitive to river regulation typology. Biological communities respond to this regulation gradient with different sensibilities and rates of response, with riparian vegetation and macroinvertebrates being the most responsive to river regulation and its gradient. Richness appears to be the best indicator for general fluvial ecological quality facing river regulation. Nevertheless, there are high correlations between the biological and functional diversity indices of different biological groups, which denotes biological connections indicative of a cascade of effects leading to an indirect influence of river regulation even on non-responsive facets of communities' biological and functional diversities. These results highlight the necessary holistic perspective of the fluvial system when assessing the effects of river regulation and the proposal of restoration measures.publishe

A versatile nanocarrierCubosomes, characterization, and applications

The impact of nanotechnology on the exponential growth of several research areas, particularly nanomedicine, is undeniable. The ability to deliver active molecules to the desired site could significantly improve the efficiency of medical treatments. One of the nanocarriers developed which has drawn researchers’ attention are cubosomes, which are nanosized dispersions of lipid bicontinuous cubic phases in water, consisting of a lipidic interior and aqueous domains folded in a cubic lattice. They stand out due to their ability to incorporate hydrophobic, hydrophilic, and amphiphilic compounds, their tortuous internal configuration that provides a sustained release, and the capacity to protect and safely deliver molecules. Several approaches can be taken to prepare this structure, as well as different lipids like monoolein or phytantriol. This review paper describes the different methods to prepare nanocarriers. As it is known, the physicochemical properties of nanocarriers are very important, as they influence their pharmacokinetics and their ability to incorporate and deliver active molecules. Therefore, an extensive characterization is essential to obtain the desired effect. As a result, we have extensively described the most common techniques to characterize cubosomes, particularly nanocarriers. The exceptional properties of the cubosomes make them suitable to be used in several applications in the biomedical field, from cancer therapeutics to imaging, which will be described. Taking in consideration the outstanding properties of cubosomes, their application in several research fields is envisaged.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit, by Marie Skłodowska Curie grant (MSCA-RISE; FODIAC; 778388) and by European Regional Development Fund (ERDF) through the Competitiveness factors Operational program—Norte 2020, COMPETE and National Funds through the FCT—under the project AgriFood XXI (NORTE- 01-0145-FEDER-000041). J.L.P. acknowledge the Instituto de Salud Carlos III for a “Sara Borrell” grant (CD19/00250), cofounded by European Social Fund (“El FSE invierte en futuro”). C.J.O.F. acknowledge the FCT for the grant SFRH/149/BD/14199/2019.info:eu-repo/semantics/publishedVersio

The impact of COVID-19 on cancer care and oncology clinical research: an experts' perspective

The coronavirus disease-19 (COVID-19) pandemic promises to have lasting impacts on cancer clinical trials that could lead to faster patient access to new treatments. In this article, an international panel of oncology experts discusses the lasting impacts of the pandemic on oncology clinical trials and proposes solutions for clinical trial stakeholders, with the support of recent data on worldwide clinical trials collected by IQVIA. These lasting impacts and proposed solutions encompass three topic areas. Firstly, acceleration and implementation of new operational approaches to oncology trials with patient-centric, fully decentralized virtual approaches that include remote assessments via telemedicine and remote devices. Geographical differences in the uptake of remote technology, including telemedicine, are discussed in the article, focusing on the impact of the local adoption of new operational approaches. Secondly, innovative clinical trials. The pandemic has highlighted the need for new trial designs that accelerate research and limit risks and burden for patients while driving optimization of clinical trial objectives and endpoints, while testing is being minimized. Areas of considerations for clinical trial stakeholders are discussed in detail. In addition, the COVID-19 pandemic has exposed the underrepresentation of minority groups in clinical trials; the approach for oncology clinical trials to improve generalizability of efficacy and outcomes data is discussed. Thirdly, a new problem-focused collaborative framework between oncology trial stakeholders, including decision makers, to leverage and further accelerate the innovative approaches in clinical research developed during the COVID-19 pandemic. This could shorten timelines for patient access to new treatments by addressing the cultural and technological barriers to adopting new operational approaches and innovative clinical trials. The role of the different stakeholders is described, with the aim of making COVID-19 a catalyst for positive change in oncology clinical research and eventually in cancer care

Low surface expression of B7-1 (CD80) is an immunoescape mechanism of colon carcinoma

Artificially enforced expression of CD80 (B7-1) and CD86 (B7-2) on tumor cells renders them more immunogenic by triggering the CD28 receptor on T cells. The enigma is that such B7s interact with much higher affinity with CTLA-4 (CD152), an inhibitory receptor expressed by activated T cells. We show that unmutated CD80 is spontaneously expressed at low levels by mouse colon carcinoma cell lines and other transplantable tumor cell lines of various tissue origins. Silencing of CD80 by interfering RNA led to loss of tumorigenicity of CT26 colon carcinoma in immunocompetent mice, but not in immunodeficient Rag-/- mice. CT26 tumor cells bind CTLA-4Ig, but much more faintly with a similar CD28Ig chimeric protein, thus providing an explanation for the dominant inhibitory effects on tumor immunity displayed by CD80 at that expression level. Interestingly, CD80-negative tumor cell lines such as MC38 colon carcinoma and B16 melanoma express CD80 at dim levels during in vivo growth in syngeneic mice. Therefore, low CD80 surface expression seems to give an advantage to cancer cells against the immune system. Our findings are similar with the inhibitory role described for the dim CD80 expression on immature dendritic cells, providing an explanation for the low levels of CD80 expression described in various human malignancies

House dust fungal communities’ characterization: a double take on the six by sixty by six (6 × 60 × 6) project

Fungi are a group of microbes that are found with particular incidence in the indoor environment. Their direct toxicity or capability of generating toxic compounds has been associated with a large number of adverse health effects, such as infectious diseases and allergies. Given that in modern society people spend a large part of their time indoors; fungal communities’ characterization of this environmental compartment assumes paramount importance in the comprehension of health effects. House dustThis work was supported by European Funds through COMPETE and by National Funds through the Portuguese Science Foundation (FCT) within project PEstOE/SAU/UI0709/2014. Ana C. A. Sousa and Sónia D. Coelho acknowledge FCT for the grants SFRH/BPD/65884/2009 and SFRH/ BD/78168/2011 (supported by funding from the Human Potential Operational Programme POPH, inscribed in the National Strategic Reference Framework and partially subsidized by the European Social Fund).info:eu-repo/semantics/publishedVersio

Transcriptome and Proteome of Fish-Pathogenic Streptococcus agalactiae Are Modulated by Temperature

Streptococcus agalactiae is one of the most important pathogens associated with streptococcosis outbreaks in Nile tilapia farms worldwide. High water temperature (above 27°C) has been described as a predisposing factor for the disease in fish. At low temperatures (below 25°C), fish mortalities are not usually observed in farms. Temperature variation can modulate the expression of genes and proteins involved in metabolism, adaptation, and bacterial pathogenicity, thus increasing or decreasing the ability to infect the host. This study aimed to evaluate the transcriptome and proteome of a fish-pathogenic S. agalactiae strain SA53 subjected to in vitro growth at different temperatures using a microarray and label-free shotgun LC-HDMSE approach. Biological triplicates of isolates were cultured in BHIT broth at 22 or 32°C for RNA and protein isolation and submitted for transcriptomic and proteomic analyses. In total, 1,730 transcripts were identified in SA53, with 107 genes being differentially expressed between the temperatures evaluated. A higher number of genes related to metabolism, mainly from the phosphotransferase system (PTS) and ATP-binding cassette (ABC) transport system, were upregulated at 32°C. In the proteome analysis, 1,046 proteins were identified in SA53, of which 81 were differentially regulated between 22 and 32°C. Proteins involved in defense mechanisms, lipid transport and metabolism, and nucleotide transport and metabolism were upregulated at 32°C. A higher number of interactions were observed in proteins involved in nucleotide transport and metabolism. We observed a low correlation between the transcriptome and proteome datasets. Our study indicates that the transcriptome and proteome of a fish-adapted S. agalactiae strain are modulated by temperature, particularly showing differential expression of genes/proteins involved in metabolism, virulence factors, and adaptation

Blocking TLR7- and TLR9-mediated IFN-α Production by Plasmacytoid Dendritic Cells Does Not Diminish Immune Activation in Early SIV Infection

Persistent production of type I interferon (IFN) by activated plasmacytoid dendritic cells (pDC) is a leading model to explain chronic immune activation in human immunodeficiency virus (HIV) infection but direct evidence for this is lacking. We used a dual antagonist of Toll-like receptor (TLR) 7 and TLR9 to selectively inhibit responses of pDC but not other mononuclear phagocytes to viral RNA prior to and for 8 weeks following pathogenic simian immunodeficiency virus (SIV) infection of rhesus macaques. We show that pDC are major but not exclusive producers of IFN-α that rapidly become unresponsive to virus stimulation following SIV infection, whereas myeloid DC gain the capacity to produce IFN-α, albeit at low levels. pDC mediate a marked but transient IFN-α response in lymph nodes during the acute phase that is blocked by administration of TLR7 and TLR9 antagonist without impacting pDC recruitment. TLR7 and TLR9 blockade did not impact virus load or the acute IFN-α response in plasma and had minimal effect on expression of IFN-stimulated genes in both blood and lymph node. TLR7 and TLR9 blockade did not prevent activation of memory CD4+ and CD8+ T cells in blood or lymph node but led to significant increases in proliferation of both subsets in blood following SIV infection. Our findings reveal that virus-mediated activation of pDC through TLR7 and TLR9 contributes to substantial but transient IFN-α production following pathogenic SIV infection. However, the data indicate that pDC activation and IFN-α production are unlikely to be major factors in driving immune activation in early infection. Based on these findings therapeutic strategies aimed at blocking pDC function and IFN-α production may not reduce HIV-associated immunopathology. © 2013 Kader et al