Human, mouse, fly and yeast GUP1 orthologues in Candida albicans

Dissertação de mestrado em Genética MolecularIn C. albicans the deletion of GUP1 causes drastic alterations at both morphologic and physiologic levels, compromising several virulence factors, such as (i) peculiar colony morphology, (ii) filamentous growth defects, (iii) impairment on the ability of cells to adhere, invade and form biofilms, and (iv) resistance to the antifungal agents. Gup1 is a yeast O-acyltransferase with close homologues in higher Eukaryotes, which has been implicated in the regulation of the Hedgehog pathway in mouse. Based on that, a primarily aim for the present work concerned the creation of a strain collection of C. albicans Δgup1 mutant harboring available GUP1 homologues from well known model organisms/higher Eukaryotes: yeast Saccharomyces cerevisiae (ScGUP1), Fly Drosophila melanogaster (DmGUP1), Mouse Mus musculus (MmGUP1), and Man Homo sapiens (HsGUP1). These transformed C. albicans strains were tested for several virulence factors, such morphology and differentiation, agar adherence and invasion capacities, and resistance to antifungals agents. Another important purpose of this study was to obtain for the first time a detailed characterization of the proteins secreted onto the C. albicans extracellular matrix. For that we imitated biofilm formation on plate and compare the wt, Δgup1 mutant and transformants. Our results suggest an apparent initial development of hyphae/pseudohyphae cells in liquid cultures, when the GUP1 homologues are inserted into Δgup1 mutant strain, yet the same strains are unable to give continuity to that differentiation. In agreement, on solid cultures those transformant strains, though displaying a colony morphology closer to wt strain were powerless to differentiate into hyphas. Moreover all the CaΔgup1GUP1 homologues showed the same impairment displayed by Δgup1mutant strain, in what regards adherence and agar invasion capacities. Concerning antifungal resistance, it seems that only DmGUP1 gene complements Δgup1 mutation. Additionally, the analysis of Δgup1 mutant extracellular matrix protein profile revealed lack of some proteins, presence of other that were not on wt profile and different concentration of some of the common proteins, suggesting that the lack of Gup1p triggers C. albicans to produce or to excrete distinct proteins. In conclusion, our results open a field of options to future research, mainly regarding proteomics of ECM and its relation to the virulence of C. albicans.A deleção do gene GUP1 em C. albicans provoca-lhe alterações drásticas tanto a nível morfológico como fisiológico, comprometendo vários factores de virulência tais como a capacidade de desenvolver hifas, a capacidade de aderir invadir e formar biofilmes, alterações de morfologia nas colónias bem com um aumento da resistência a antifúngicos. A proteína Gup1p é uma O-aciltransferase presente em leveduras e que partilha uma elevada homologia com os eucariotas superiores, tendo sido descrita como essencial na regulação da via Hedgehog, em ratinhos. Com base nestes conhecimentos, foi proposto como 1° objectivo deste trabalho a criação de uma colecção de estirpes de C. albicans mutada no gene GUP1 complementada com vários homólogos do GUP1 de eucariotas superiores: levedura Saccharomyces cerevisiae (ScGUP1), mosca Drosophila melanogaster (DmGUP1), ratinho - Mus musculus (MmGUP1), humano -Homo sapiens (HsGUP1). Nestas novas estirpes foram testados vários factores de virulência: morfologia, capacidade de diferenciação em filamentos, aderência e invasão ao agar, e ainda a resistência a antifúngicos. Um outro objectivo importante deste trabalho consistiu na caracterização detalhada das proteínas secretadas para a matriz extracelular da C. albicans. Para tal simulou-se a formação de biofilmes em placas comparando as estirpes wt e mutada no GUP1. Os resultados obtidos mostram que os vários homólogos do GUP1 quando inseridos na estirpe mutada tiveram a capacidade de iniciar diferenciação em hifas/pseudohifas, em meio líquido. No entanto, não foram aptas para dar continuidade à referida diferenciação. Do mesmo modo, em meio sólido as colónias destas estirpes foram morfologicamente mais semelhantes à wt que ao mutante GUP1, sem no entanto apresentarem hifas. Para além disso, nenhum dos transformantes mostrou capacidade de aderir ou invadir o agar. Quando testada a sua resistência a antifúngicos apenas a estirpe complementada com o DmGUP1 (da mosca) reverteu o fenótipo do mutante GUP1. Por fim, a análise das proteínas da matriz extracelular da wt e do mutante GUP1 revelou perfis proteicos muito distintos, presença de diferentes proteínas e também em diferentes quantidades, o que sugere que a ausência do GUP1 provoca uma alterada excreção de proteínas

Near-Optimal Detection of CE-OFDM Signals with High Power Efficiency via GAMP-based Receivers

Proceeding of: 2022 IEEE Globecom Workshops (GC Wkshps), Rio de Janeiro, Brazil, 4-8 December 2022A quasi-optimum receiver based on the generalized approximate message passing (GAMP) concept is proposed for constant envelope orthogonal frequency division multiplexing (CE-OFDM) signals. Large modulation index results in large power efficiency for CE-OFDM, but the phase modulator introduces nonlinear distortion effects, precluding good performance for a simple phase detector. Our simulation results show that the GAMP receiver can achieve quasi-optimum performance and it can outperform the linear OFDM and CE-OFDM with phase detectors, for both additive white Gaussian noise (AWGN) and frequency selective channels.This work received funding from the European Union (EU) Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie ETN TeamUp5G, grant agreement No. 813391, the Spanish National Project IRENE-EARTH (PID2020-115323RB-C33) (MINECO/AEI/FEDER, UE) and Portuguese FCT Instituto de Telecomunicaçoes project UIDB/50008/2020

Chondral lesion of the patella in a junior football player: chondral-only fragment fixation

Chondral lesions of the patella are a relatively common pathology that can cause pain and impairment in active young individuals. These lesions can have lifelong consequences, with increased risk of osteoarthritis in some cases. Surgical excision of medium to large fragments is associated with poor long-term outcomes. There is a general belief that fixation of purely chondral lesions in the absences of subchondral bone is associated with low integration rates, however recent case reports and basic science studies have been challenging this belief. In this case report, we describe a young patient with a chondral lesion of the patella and a fragment without subchondral bone, who underwent open fragment reduction and fixation using bioabsorbable pins. The patient had a successful outcome with significant fast improvement in pain and function and full return to practice. This case highlights the potential benefits of conservative surgical management with preservation of the native cartilage, even in the absence of subchondral bone, especially in the young active patient.

Yeasts, the man’s best friend

1. Introduction In most cultures, bread making depends on a fermentation step. The flour leavening ability was, at first, most probably dependent on spontaneous fermentation. It became a controlled process by the maintenance of fresh innocula from one preparation to the next and this kind of environmental constraints eventually generated a particular type of yeast and bacteria biodiversity, adapted to ferment a certain brand of flour mixture, yielding specific organoleptic characteristics to the dough. Nowadays, although the baking industry generally uses commercially available strains of Saccharomyces cerevisiae for bread making, some types of bread are still prepared using dough carried over from previous makings as a starter. This trend decreased worldwide bread diversity and the cultural values associated, simultaneously increasing the dependence of local producers on world-scale yeast producers. Sustainability demands assessing yeast biodiversity, as well as devising simple and cheap methods for maintaining dough and multiply yeast.Este trabalho é financiado por Fundos FEDER através do Programa Operacional Factores de Competitividade – COMPETE e por Fundos Nacionais através da FCT – Fundação para a Ciência e a Tecnologia no âmbito do projecto PEst-C/BIA/UI4050/201

Exploiting Leishmania—Primed Dendritic Cells as Potential Immunomodulators of Canine Immune Response

Funding Information: This research was funded by the FCT-Foundation for Science and Technology, I.P., through research grants PTDC/CVT-CVT/28908/2017 (http://doi.org/10.54499/PTDC/CVT-CVT/28908/2017) and PTDC/CVT-CVT/0228/2020 (http://doi.org/10.54499/PTDC/CVT-CVT/0228/2020) and by national funds within the scope of Centro de Investigação Interdisciplinar em Sanidade Animal (CIISA, UIDB/00276/2020), AL4AnimalS—LA/P/0059/2020, Global Health and Tropical Medicine (GHTM, UID/04413/2020), and LA-REAL—LA/P/0117/2020. Publisher Copyright: © 2024 by the authors.Dendritic cells (DCs) capture pathogens and process antigens, playing a crucial role in activating naïve T cells, bridging the gap between innate and acquired immunity. However, little is known about DC activation when facing Leishmania parasites. Thus, this study investigates in vitro activity of canine peripheral blood-derived DCs (moDCs) exposed to L. infantum and L. amazonensis parasites and their extracellular vesicles (EVs). L. infantum increased toll-like receptor 4 gene expression in synergy with nuclear factor κB activation and the generation of pro-inflammatory cytokines. This parasite also induced the expression of class II molecules of major histocompatibility complex (MHC) and upregulated co-stimulatory molecule CD86, which, together with the release of chemokine CXCL16, can attract and help in T lymphocyte activation. In contrast, L. amazonensis induced moDCs to generate a mix of pro- and anti-inflammatory cytokines, indicating that this parasite can establish a different immune relationship with DCs. EVs promoted moDCs to express class I MHC associated with the upregulation of co-stimulatory molecules and the release of CXCL16, suggesting that EVs can modulate moDCs to attract cytotoxic CD8+ T cells. Thus, these parasites and their EVs can shape DC activation. A detailed understanding of DC activation may open new avenues for the development of advanced leishmaniasis control strategies.publishersversionpublishe

Exploiting <i>Leishmania</i>—Primed Dendritic Cells as Potential Immunomodulators of Canine Immune Response

Dendritic cells (DCs) capture pathogens and process antigens, playing a crucial role in activating naïve T cells, bridging the gap between innate and acquired immunity. However, little is known about DC activation when facing Leishmania parasites. Thus, this study investigates in vitro activity of canine peripheral blood-derived DCs (moDCs) exposed to L. infantum and L. amazonensis parasites and their extracellular vesicles (EVs). L. infantum increased toll-like receptor 4 gene expression in synergy with nuclear factor κB activation and the generation of pro-inflammatory cytokines. This parasite also induced the expression of class II molecules of major histocompatibility complex (MHC) and upregulated co-stimulatory molecule CD86, which, together with the release of chemokine CXCL16, can attract and help in T lymphocyte activation. In contrast, L. amazonensis induced moDCs to generate a mix of pro- and anti-inflammatory cytokines, indicating that this parasite can establish a different immune relationship with DCs. EVs promoted moDCs to express class I MHC associated with the upregulation of co-stimulatory molecules and the release of CXCL16, suggesting that EVs can modulate moDCs to attract cytotoxic CD8+ T cells. Thus, these parasites and their EVs can shape DC activation. A detailed understanding of DC activation may open new avenues for the development of advanced leishmaniasis control strategies

6G Technology Overview (third edition)

6G aims to address diverse, often competing needs, such as vastly increased data rates, a massive scale of communicating devices, energy efficiency, and the simultaneous demand for both high data rates and low communication latency. The White Paper covers several groundbreaking technologies pivotal for 6G evolution, such as terahertz frequencies, 6G radio access, integrated sensing and communication, non-terrestrial networks and more. For each technology, the White Paper offers background information, explains its relevance to 6G, presents key problems, and provides a thorough review of the current state of the art. The paper emphasises that while the listed technologies form the foundation for 6G, the landscape will continuously evolve. Subsequent versions of the white paper will spotlight new technologies and integrate them into a cohesive system. The “6G Technology Overview” concludes that 6G, with its vast potential, aims not only to meet the diverse requirements of novel use cases but also to ensure sustainability, user-friendliness, and ease of service deployment