175 research outputs found
Effect of Chromatographic Conditions on Supercoiled Plasmid DNA Stability and Bioactivity
Funding: This work was supported by FEDER funds through the POCI—COMPETE 2020 Operational Programme Competitiveness and Internationalization in Axis I—Strengthening research, technological development, and innovation (Project POCI-01-0145-FEDER-007491), and National Funds by FCT Foundation for Science and Technology (Project UID/Multi /00709/2013). This work was also developed within the scope of the project CICECO-Aveiro Institute of Materials, FCT Ref. UID/CTM/50011/2019, financed by national funds through the FCT/MCTES. G.M. Azevedo acknowledges the support and fellowship of Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/203482/2014-0). J.F.A. Valente acknowledges the PhD fellowship (Ref SFRH/BD/96809/2013) from FCT.Acknowledgments: The authors would like to thank Thomas Roberts for providing the pcDNA3–FLAG–p53
construct through Addgene, ref: 10838.The dysfunction of the tumor suppressor gene TP53 has been associated with the pathogenesis of the majority of the cases of cancer reported to date, leading the cell to acquire different features known as the cancer hallmarks. In normal situations, the protein p53 protects the cells against tumorigenesis. By detecting metabolic stress or DNA damage in response to stress, p53 can lead the cell to senescence, autophagy, cell cycle arrest, DNA repair, and apoptosis. Thus, in the case of p53 mutations, it is reasonable to assume that the reestablishment of its function, may restrain the proliferation of cancer cells. The concept of cancer gene therapy can be based on this assumption, and suitable biotechnological approaches must be explored to assure the preparation of gene-based biopharmaceuticals. Although numerous procedures have already been established to purify supercoiled plasmid DNA (sc pDNA), the therapeutic application is highly dependent on the biopharmaceutical’s activity, which can be affected by the chromatographic conditions used. Thus, the present work aims at comparing quality and in vitro activity of the supercoiled (sc) isoform of the p53 encoding plasmid purified by three different amino acids-based chromatographic strategies, involving histidine–agarose, arginine–macroporous, and histidine–monolith supports. The B-DNA topology was maintained in all purified pDNA samples, but their bioactivity, related to the induction of protein p53 expression and apoptosis in cancer cells, was higher with arginine–macroporous support, followed by histidine–monolith and histidine–agarose. Despite the purity degree of 92% and recovery yield of 43% obtained with arginine–macroporous, the sc pDNA sample led to a higher expression level of the therapeutic p53 protein (58%) and, consequently, induced a slightly higher apoptotic effect (27%) compared with sc pDNA samples obtained with histidine–monolithic support (26%) and histidine–agarose support (24%). This behavior can be related to the mild chromatographic conditions used with arginine–macroporous support, which includes the use of low salt concentrations, at neutral pH and lower temperatures, when compared to the high ionic strength of ammonium sulfate and acidic pH used with histidine-based supports. These results can contribute to field of biopharmaceutical preparation, emphasizing the need to control several experimental conditions while adapting and selecting the methodologies that enable the use of milder conditions as this can have a significant impact on pDNA stability and biological activity.info:eu-repo/semantics/publishedVersio
Quasivariational solutions for first order quasilinear equations with gradient constraint
We prove the existence of solutions for an evolution quasi-variational
inequality with a first order quasilinear operator and a variable convex set,
which is characterized by a constraint on the absolute value of the gradient
that depends on the solution itself. The only required assumption on the
nonlinearity of this constraint is its continuity and positivity. The method
relies on an appropriate parabolic regularization and suitable {\em a priori}
estimates. We obtain also the existence of stationary solutions, by studying
the asymptotic behaviour in time. In the variational case, corresponding to a
constraint independent of the solution, we also give uniqueness results
Coexistence or Separation of the Superconducting, Antiferromagnetic, and Paramagnetic Phases in Quasi One-Dimensional (TMTSF)2PF6 ?
We report on experimental studies of the character of phase transitions in
the quasi-1D organic compound (TMTSF)2PF6 in the close vicinity of the borders
between the paramagnetic metal PM, antiferromagnetic insulator AF, and
superconducting SC states. In order to drive the system through the phase
border P_0(T_0), the sample was maintained at fixed temperature T and pressure
P, whereas the critical pressure P_0 was tuned by applying the magnetic field
B. In this approach, the magnetic field was used (i) for tuning (P-P_0), and
(ii) for identifying the phase composition (due to qualitatively different
magnetoresistance behavior in different phases). Experimentally, we measured
R(B) and its temperature dependence R(B,T) in the pressure range (0 - 1)GPa.
Our studies focus on the features of the magnetoresistance at the phase
transition between the PM and AF phases, in the close vicinity to the
superconducting transition at T~1K. We found pronounced history effects arising
when the AF/PM phase border is crossed by sweeping the magnetic field: the
resistance depends on a trajectory which the system arrives at a given point of
the P-B-T phase space. In the transition from the PM to AF phase, the features
of the PM phase extends well into the AF phase. At the opposite transition from
the AF to PM phase, the features of the AF phase are observed in the PM phase.
These results evidence for a macroscopically inhomogeneous state, which
contains macroscopic inclusions of the minority phase. When the system is
driven away from the transition, the homogeneous state is restored; upon a
return motion to the phase boundary, no signatures of the minority phase are
observed up to the very phase boundary.Comment: 10 figures, 23 page
How brains make decisions
This chapter, dedicated to the memory of Mino Freund, summarizes the Quantum
Decision Theory (QDT) that we have developed in a series of publications since
2008. We formulate a general mathematical scheme of how decisions are taken,
using the point of view of psychological and cognitive sciences, without
touching physiological aspects. The basic principles of how intelligence acts
are discussed. The human brain processes involved in decisions are argued to be
principally different from straightforward computer operations. The difference
lies in the conscious-subconscious duality of the decision making process and
the role of emotions that compete with utility optimization. The most general
approach for characterizing the process of decision making, taking into account
the conscious-subconscious duality, uses the framework of functional analysis
in Hilbert spaces, similarly to that used in the quantum theory of
measurements. This does not imply that the brain is a quantum system, but just
allows for the simplest and most general extension of classical decision
theory. The resulting theory of quantum decision making, based on the rules of
quantum measurements, solves all paradoxes of classical decision making,
allowing for quantitative predictions that are in excellent agreement with
experiments. Finally, we provide a novel application by comparing the
predictions of QDT with experiments on the prisoner dilemma game. The developed
theory can serve as a guide for creating artificial intelligence acting by
quantum rules.Comment: Latex file, 20 pages, 3 figure
Immunophenotype of gastric tumors unveils a pleiotropic role of regulatory T cells in tumor development
Simple SummaryThe role of regulatory T cells (Tregs) in gastric cancer (GC) is still controversial and poorly understood. GC patients have increased numbers of Tregs in peripheral blood and among tumor infiltrating lymphocytes; however, their prognostic value depends on specific tumor features (e.g., tumor location and/or microsatellite instability status). We found that Tregs might induce membrane expression of IL2R alpha in intestinal-type GC cells, which associates with MAPK signaling pathway activation and spheroid growth. Moreover, Tregs accumulate at early steps of intestinal-type GCs progression, when tumors are starting to grow through the stomach wall, and do not present vascular and perineural invasion. Our findings suggest a novel non-immunosuppressive role of Treg cells in intestinal-type GC, which may unlock novel therapeutic immuno-oncology strategies for intestinal-type GC or other tumors with similar immune context.Gastric cancer (GC) patients display increased regulatory T cell (Tregs) numbers in peripheral blood and among tumor-infiltrating lymphocytes. Nevertheless, the role of Tregs in GC progression remains controversial. Here, we sought to explore the impact of Tregs in GCs with distinct histology, and whether Tregs can directly influence tumor cell behavior and GC development. We performed a comprehensive immunophenotyping of 82 human GC cases, through an integrated analysis of multispectral immunofluorescence detection of T cells markers and patient clinicopathological data. Moreover, we developed 3D in vitro co-cultures with Tregs and tumor cells that were followed by high-throughput and light-sheet imaging, and their biological features studied with conventional/imaging flow cytometry and Western blotting. We showed that Tregs located at the tumor nest were frequent in intestinal-type GCs but did not associate with increased levels of effector T cells. Our in vitro results suggested that Tregs preferentially infiltrated intestinal-type GC spheroids, induced the expression of IL2R alpha and activation of MAPK signaling pathway in tumor cells, and promoted spheroid growth. Accumulation of Tregs in intestinal-type GCs was increased at early stages of the stomach wall invasion and in the absence of vascular and perineural invasion. In this study, we proposed a non-immunosuppressive mechanism through which Tregs might directly modulate GC cells and thereby promote tumor growth. Our findings hold insightful implications for therapeutic strategies targeting intestinal-type GCs and other tumors with similar immune context.MTG4Molecular tumour pathology - and tumour genetic
INFLUÊNCIA DE ELEMENTOS DO CLIMA NO DESPERDÍCIO DE ENERGIA EM UM SISTEMA DE IRRIGAÇÃO POR ASPERSÃO DE ALTA PRESSÃO
Efeito da aplicação de fungicidas no controle de doenças foliares de arroz irrigado e sua relação com o rendimento industrial
Strange particle production in proton-proton collisions at TeV with ALICE at the LHC
The production of mesons containing strange quarks (K, ) and both
singly and doubly strange baryons (, Anti-, and
+Anti-) are measured at central rapidity in pp collisions at
= 0.9 TeV with the ALICE experiment at the LHC. The results are
obtained from the analysis of about 250 k minimum bias events recorded in 2009.
Measurements of yields (dN/dy) and transverse momentum spectra at central
rapidities for inelastic pp collisions are presented. For mesons, we report
yields () of 0.184 0.002 stat. 0.006 syst. for K and
0.021 0.004 stat. 0.003 syst. for . For baryons, we find
= 0.048 0.001 stat. 0.004 syst. for , 0.047
0.002 stat. 0.005 syst. for Anti- and 0.0101 0.0020 stat.
0.0009 syst. for +Anti-. The results are also compared with
predictions for identified particle spectra from QCD-inspired models and
provide a baseline for comparisons with both future pp measurements at higher
energies and heavy-ion collisions.Comment: 33 pages, 21 captioned figures, 10 tables, authors from page 28,
published version, figures at
http://aliceinfo.cern.ch/ArtSubmission/node/387
Two-pion Bose-Einstein correlations in central Pb-Pb collisions at = 2.76 TeV
The first measurement of two-pion Bose-Einstein correlations in central Pb-Pb
collisions at TeV at the Large Hadron Collider is
presented. We observe a growing trend with energy now not only for the
longitudinal and the outward but also for the sideward pion source radius. The
pion homogeneity volume and the decoupling time are significantly larger than
those measured at RHIC.Comment: 17 pages, 5 captioned figures, 1 table, authors from page 12,
published version, figures at
http://aliceinfo.cern.ch/ArtSubmission/node/388
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