No induction of antimicrobial resistance in Staphylococcus aureus and Listeria monocytogenes during continuous exposure to eugenol and citral

The aim of this study was to evaluate the adaptation response of Staphylococcus aureus, methicillin-resistant S.aureus (MRSA), and Listeria monocytogenes to the essential oil (EO), eugenol, and citral. The minimum inhibitory concentration of eugenol and citral was determined by agar dilution and microdilution. Adaptation to eugenol and citral was done by sequential exposure of the pathogens to increasing concentrations of the essential oils. The M2-A9 standard was used to determine the antibiotic susceptibility. The effect of eugenol and citral on the adherence ability was evaluated by the crystal violet assay. The impact of adaptation to eugenol on virulence was estimated using the Galleria mellonella model. No development of resistance to the components and antibiotics was observed in the adapted cells of S.aureus, MRSA, and L.monocytogenes. Eugenol and citral at subinhibitory concentration reduced the bacterial adherence. Adaptation to subinhibitory concentration of eugenol affected the virulence potential of S.aureus, MRSA, and L.monocytogenes. Eugenol and citral do not pose a risk of resistance development in a continuous mode of use. These EO components showed a high efficacy as antistaphylococcal and antilisterial biofilm agents. Adaptation at subinhibitory concentration of eugenol protected the larvae against listerial and staphylococcal infection.FCT - Fundacao para a Ciencia e Tecnologia [PEst-OE/EQB/LA0023/2013]info:eu-repo/semantics/publishedVersio

Untangling the immune basis of disease susceptibility

© 2020 Copyright Ribeiro and Graca. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are creditedInteractions between immune cell receptors and proteins that determine disease susceptibility shed light on how different arms of the immune system are involved in three viral infections and Crohn's disease.info:eu-repo/semantics/publishedVersio

Judicial Review of the Legislative Process in Brazil

Judicial review of the legislative process has been a controversial topic in the case law of the Federal Supreme Court in Brazil. Issues regarding enacted statutes are not as controversial as those involving pending processes. On one hand, the Court has been scrutinising cases based on procedural legislative rules that are enshrined in the Constitution. On the other, the Court has been refusing to examine procedures based on provisions not enshrined in the Constitution, such as the internal ordinances of the parliament. In the former situation, the Court sees itself compelled to perform a kind of prior control of constitutionality. In the latter, it states that evaluating whether the process abides by the parliament’s own rules is an internal political (interna corporis) task, not a legal one. In this article, I argue that adherence to rules that govern the legislative process, regardless of their status, is not a matter of political discretionary choices, but a matter of compliance with the rule of law. Therefore, the Brazilian Federal Supreme Court should abandon the prior control of constitutionality rationale, and it should review pending legislative processes including those based on the internal ordinances of the parliament

Identification of Regulatory T Cells in Tolerated Allografts

Induction of transplantation tolerance with certain therapeutic nondepleting monoclonal antibodies can lead to a robust state of peripheral “dominant” tolerance. Regulatory CD4+ T cells, which mediate this form of “dominant” tolerance, can be isolated from spleens of tolerant animals. To determine whether there were any extra-lymphoid sites that might harbor regulatory T cells we sought their presence in tolerated skin allografts and in normal skin. When tolerated skin grafts are retransplanted onto T cell–depleted hosts, graft-infiltrating T cells exit the graft and recolonize the new host. These colonizing T cells can be shown to contain members with regulatory function, as they can prevent nontolerant lymphocytes from rejecting fresh skin allografts, without hindrance of rejection of third party skin. Our results suggest that T cell suppression of graft rejection is an active process that operates beyond secondary lymphoid tissue, and involves the persistent presence of regulatory T cells at the site of the tolerated transplant

Co-receptor and co-stimulation blockade for mixed chimerism and tolerance without myelosuppressive conditioning

BACKGROUND: A major challenge in the application of marrow transplantation as a route to immunological tolerance of a transplanted organ is to achieve hematopoietic stem cell (HSC) engraftment with minimal myelosuppressive treatments. RESULTS: We here describe a combined antibody protocol which can achieve long-term engraftment with clinically relevant doses of MHC-mismatched bone marrow, without the need for myelosuppressive drugs. Although not universally applicable in all strains, we achieved reliable engraftment in permissive strains with a two-stage strategy: involving first, treatment with anti-CD8 and anti-CD4 in advance of transplantation; and second, treatment with antibodies targeting CD4, CD8 and CD40L (CD154) at the time of marrow transplantation. Long-term mixed chimerism through co-receptor and co-stimulation blockade facilitated tolerance to donor-type skin grafts, without any evidence of donor-antigen driven regulatory T cells. CONCLUSION: We conclude that antibodies targeting co-receptor and co-stimulatory molecules synergise to enable mixed hematopoietic chimerism and central tolerance, showing that neither cytoreductive conditioning nor 'megadoses' of donor bone marrow are required for donor HSC to engraft in permissive strains

Maturation and phenotypic heterogeneity of human CD4+ regulatory T cells from birth to adulthood and after allogeneic stem cell transplantation

Copyright © 2021 Matos, Hirakawa, Alho, Neleman, Graca and Ritz. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.CD4+ Regulatory T cells (Treg) play a critical role in maintaining immune homeostasis. Various Treg subsets have been identified, however the heterogeneity of Treg subpopulations during development remains uncharacterized. Using mass cytometry we obtained single cell data on expression of 35 functional markers to examine the heterogeneity of Treg cells at birth and in adults. Unsupervised clustering algorithms FlowSOM and ACCENSE were used to quantify Treg heterogeneity. As expected, Treg in umbilical cord blood were predominately naïve while Treg in adult blood were predominately central memory and effector memory cells. Although umbilical cord blood Treg are mostly naïve cells, we observed multiple phenotypic Treg subsets in cord blood. Nevertheless, peripheral blood in adults contained higher percentages of Treg and the heterogeneity of Treg was significantly increased in adults. We also studied Treg heterogeneity throughout a 2-year period after allogeneic hematopoietic stem cell transplantation (alloHSCT) and in patients with chronic graft-versus-host disease (cGVHD). Treg heterogeneity recovered rapidly after alloHSCT and gradually increased in the first two years post-transplant. However, patients with cGVHD had significantly fewer distinct Treg subpopulations, proposing a correlation between a disrupted Treg heterogeneity and cGVHD. Our study is the first to compare human Treg heterogeneity at birth, in healthy adults and in patients after alloHSCT with and without cGVHD. This approach to characterize Treg heterogeneity based on expression of a large panel of functional markers may enable future studies to identify specific Treg defects that contribute to immune dysfunction.This work was supported by NIH grant P01CA229092, EADV Research Fellowship, Rene-Touraine Fellowship and a generous contribution from the Fundação para a Ciência e a Tecnologia (FCT), FCT SFRH/BD/98980/2013info:eu-repo/semantics/publishedVersio

Carbon Nanotubes Modified Oil-based Nanofluids: Electrical Conduction Mechanism Analysis

This paper reports research on the electrical conduction mechanisms of nanofluids, formed by adding carbon nanotube (CNT) and 0.5 wt% of graphite (Gt) particles to engine oil, in the range of temperatures up to 400 K. An analysis of the dc conductivity of nanofluids versus CNT concentration and temperature, have shown a percolation threshold of 0.6 wt% and revealed the PTCR (Positive Temperature Coefficient of Resistivity) effect for temperatures below the critical value Tc≈350 K and concentrations above the critical percolation threshold. The ac conductivity of all samples was fitted in the range of frequency from 100 Hz to 1 MHz using the Jonscher power law, which allows for the specification of the appropriate electrical conduction mechanism using the exponent s in that law

IL-9 expression by invariant NKT cells is not imprinted during thymic development

Copyright © 2015 by The American Association of Immunologists, Inc. All rights reservedInvariant NKT (iNKT) cell thymic development can lead to distinct committed effector lineages, namely NKT1, NKT2, and NKT17. However, following identification of IL-9-producing iNKT cells involved in mucosal inflammation, their development remains unaddressed. In this study, we report that although thymic iNKT cells from naive mice do not express IL-9, iNKT cell activation in the presence of TGF-β and IL-4 induces IL-9 secretion in murine and human iNKT cells. Acquisition of IL-9 production was observed in different iNKT subsets defined by CD4, NK1.1, and neuropilin-1, indicating that distinct functional subpopulations are receptive to IL-9 polarization. Transcription factor expression kinetics suggest that regulatory mechanisms of IL-9 expression are shared by iNKT and CD4 T cells, with Irf4 and Batf deficiency deeply affecting IL-9 production. Importantly, adoptive transfer of an enriched IL-9(+) iNKT cell population leads to exacerbated allergic inflammation in the airways upon intranasal immunization with house dust mite, confirming the ability of IL-9-producing iNKT cells to mediate proinflammatory effects in vivo, as previously reported. Taken together, our data show that peripheral iNKT cells retain the capacity of shaping their function in response to environmental cues, namely TGF-β and IL-4, adopting an IL-9-producing NKT cell phenotype able to mediate proinflammatory effects in vivo, namely granulocyte and mast cell recruitment to the lungs.This work was supported by Fundação Para a Ciência e Tecnologia, Portugal Grants PTDC/SAU-TOX/114424/2009 and HMSP-ICT/0034/2013 and the Novo Nordisk/European Foundation for the Study of Diabetes (to L.G.), as well as by grants from the Fundação Para a Ciência e Tecnologia, Portugal, the European Molecular Biology Organization, the European Research Council, and the U.S. National Blood Foundation (to H.V.-F.). A.A.-D., C.F.A., D.F.-P., and M.M. were supported by fellowships from the Fundação Para a Ciência e Tecnologia, Portugal.info:eu-repo/semantics/publishedVersio

Peptidylprolyl isomerase C (Ppic) regulates invariant Natural Killer T cell (iNKT) differentiation in mice

© 2021 The Authors. European Journal of Immunology published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.Peptidyl-prolyl cis-trans isomerase C (Ppic) is expressed in several bone marrow (BM) hematopoietic progenitors and in T-cell precursors. Since the expression profile of Ppic in the hematoimmune system was suggestive that it could play a role in hematopoiesis and/or T lymphocyte differentiation, we sought to test that hypothesis in vivo. Specifically, we generated a Ppic-deficient mouse model by targeting the endogenous locus by CRISPR/Cas9 and tested the requirement of Ppic in hematopoiesis. Several immune cell lineages covering BM progenitors, lymphocyte precursors, as well as mature cells at the periphery were analyzed. While most lineages were unaffected, invariant NKT (iNKT) cells were reduced in percentage and absolute cell numbers in the Ppic-deficient thymus. This affected the most mature stages in the thymus, S2 and S3, and the phenotype was maintained at the periphery. Additionally, immature transitional T1 and T2 B lymphocytes were increased in the Ppic-deficient spleen, but the phenotype was lost in mature B lymphocytes. In sum, our data show that Ppic is dispensable for myeloid cells, platelets, erythrocytes, αβ, and γδ T lymphocytes in vivo in the steady state, while being involved in B- and iNKT cell differentiation.This work was supported by the Instituto Gulbenkian de Ciência (IGC) of the Calouste Gulbenkian Foundation, and the Portuguese National Research Council (Fundação para a Ciência e Tecnologia [FCT]) Grant PTDC/BIA-BID/30925/2017 to VCM, that also supports the salary of RSP. VCM is supported by an individual contract awarded by FCT (CEECIND/03106/2018). CVR is a PhD student of the IGC Integrative Biology and Biomedicine (IBB) PhD Program and supported by an individual FCT PhD Fellowship ref. PD/BD/139190/2018. This work had the support of the research infrastructures Congento LISBOA-01-0145-FEDER-022170 and PPBI-POCI-01-0145-FEDER-022122, both cofinanced by FCT and Lisboa2020, under PORTUGAL2020 agreement (European Regional Development Fund).info:eu-repo/semantics/publishedVersio