A Filosofia crítica segundo Gilles Deleuze

Revendo as observações de Deleuze sobre as diferenças de método entre Nietzsche-Kant e entre Nietzsche-Platão, o objetivo deste ensaio é pensar os princípios da filosofia crítica

Development of a self-diagnostic system integrated into a cyber-physical system

CONTROLAR provides Bosch with an intelligent functional testing machine used to test the correct functioning of the car radios produced. During this process, the radios are submitted to several tests, raising the problem of how the machine detects errors in several radios consecutively, making it impossible to know if the device has a problem since it has no module to see if it works correctly. This article arises from the need to find a solution to solve this problem, which was to develop a self-diagnostic system that will ensure the reliability and integrity of the cyber-physical system, passing a detailed state of the art. The development of this system was based on the design of an architecture that combines the KDT methodology with a DSL to manage and configure the tests to integrate the self-diagnostic test system into a CPS. A total of 28 test cases were performed to cover all its functionalities. The results show that all test cases passed. Therefore, the system meets all the proposed objectives.This article is a result of the project POCI-01-0247-FEDER-040130, supported by Operational Program for Competitiveness and Internationalization (COMPETE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF)

Characteristics of ash and particle emissions during bubbling fluidised bed combustion of three types of residual forest biomass

Combustion of residual forest biomass (RFB) derived from eucalypt (Eucalyptus globulus), pine (Pinus pinaster) and golden wattle (Acacia longifolia) was evaluated in a pilot-scale bubbling fluidised bed reactor (BFBR). During the combustion experiments, monitoring of temperature, pressure and exhaust gas composition has been made. Ash samples were collected at several locations along the furnace and flue gas treatment devices (cyclone and bag filter) after each combustion experiment and were analysed for their unburnt carbon content and chemical composition. Total suspended particles (TSP) in the combustion flue gas were evaluated at the inlet and outlet of cyclone and baghouse filter and further analysed for organic and elemental carbon, carbonates and 57 chemical elements. High particulate matter collection efficiencies in the range of 94-99% were observed for the baghouse, while removal rates of only 1.4-17% were registered for the cyclone. Due to the sand bed, Si was the major element in bottom ashes. Fly ashes, in particular those from eucalypt combustion, were especially rich in CaO, followed by relevant amounts of SiO2, MgO and K2O. Ash characteristics varied among experiments, showing that their inorganic composition strongly depends on both the biomass composition and combustion conditions. Inorganic constituents accounted for TSP mass fractions up to 40 wt%. Elemental carbon, organic matter and carbonates contributed to TSP mass fractions in the ranges 0.58-44%, 0.79-78% and 0.01-1.7%, respectively.publishe

Effect of industrial and domestic ash from biomass combustion, and spent coffee grounds, on soil fertility and plant growth: experiments at field conditions

An experimental study was conducted at field conditions in order to evaluate the effect of application of ash from biomass combustion on some soil fertility characteristics and plant growth. Application of 7.5 Mg ha-1 industrial fly ash (IA), domestic ash (DA), and a 50:50 mix of domestic ash (DA) and spent coffee grounds (SCG) was made in different soil parcels. Lolium perenne seeds were sown and the grown biomass was harvested and quantified after 60 days. Soil samples from each parcel were also collected after that period and characterized. Both soil and grown biomass samples were analyzed for Ca, Mg, Na, K, P, Fe, Mn, Zn, and Al contents. Soil pH was determined before and after amendment. All applications rose significantly soil pH. Domestic ash, whether combined with coffee grounds or not, proved to be efficient at supplying available macronutrients Ca, Mg, K, and P to the soil and also reducing availability of Al (more than industrial ash). However, it inhibited plant growth, even more when combined with spent coffee grounds. As regards to elemental abundance in plant tissue, both domestic ash treatments reduced Ca and enhanced Al contents, unlike industrial ash, which proved less harmful for the load applied in the soil. Hence, it was possible to conclude that application load should be a limiting factor for this management option for the studied materials.publishe

Toward the Mechanistic Understanding of Enzymatic CO2 Reduction

SFRH/BD/116515/2014 PTDC/BBB-EBB/2723/2014 UID/Multi/04378/2019 grant agreement number 810856Reducing CO2 is a challenging chemical transformation that biology solves easily, with high efficiency and specificity. In particular, formate dehydrogenases are of great interest since they reduce CO2 to formate, a valuable chemical fuel and hydrogen storage compound. The metal-dependent formate dehydrogenases of prokaryotes can show high activity for CO2 reduction. Here, we report an expression system to produce recombinant W/Sec-FdhAB from Desulfovibrio vulgaris Hildenborough fully loaded with cofactors, its catalytic characterization and crystal structures in oxidized and reduced states. The enzyme has very high activity for CO2 reduction and displays remarkable oxygen stability. The crystal structure of the formate-reduced enzyme shows Sec still coordinating the tungsten, supporting a mechanism of stable metal coordination during catalysis. Comparison of the oxidized and reduced structures shows significant changes close to the active site. The DvFdhAB is an excellent model for studying catalytic CO2 reduction and probing the mechanism of this conversion.publishersversionpublishe

Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa

A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrPC), expressed by the prion gene (prnp), into an abnormally folded isoform (PrPSc) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific - Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the "prion gene complex". Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its phenylalanine residue.publishersversionpublishe

Genetic Variants Underlying Risk of Intracranial Aneurysms: Insights from a GWAS in Portugal

Subarachnoid hemorrhage (SAH) is a life-threatening event that most frequently leads to severe disability and death. Its most frequent cause is the rupture of a saccular intracranial aneurysm (IA), which is a blood vessel dilation caused by disease or weakening of the vessel wall. Although the genetic contribution to IA is well established, to date no single gene has been unequivocally identified as responsible for IA formation or rupture. We aimed to identify IA susceptibility genes in the Portuguese population through a pool-based multistage genome-wide association study. Replicate pools were allelotyped in triplicate in a discovery dataset (100 IA cases and 92 gender-matched controls) using the Affymetrix Human SNP Array 6.0. Top SNPs (absolute value of the relative allele score difference between cases and controls |RASdiff|≥13.0%) were selected for technical validation by individual genotyping in the discovery dataset. From the 101 SNPs successfully genotyped, 99 SNPs were nominally associated with IA. Replication of technically validated SNPs was conducted in an independent replication dataset (100 Portuguese IA cases and 407 controls). rs4667622 (between UBR3 and MYO3B), rs6599001 (between SCN11A and WDR48), rs3932338 (214 kilobases downstream of PRDM9), and rs10943471 (96 kilobases upstream of HTR1B) were associated with IA (unadjusted allelic chi-square tests) in the datasets tested (discovery: 6.84E-04≤P≤1.92E-02, replication: 2.66E-04≤P≤2.28E-02, and combined datasets: 6.05E-05≤P≤5.50E-04). Additionally, we confirmed the known association with IA of rs1333040 at the 9p21.3 genomic region, thus validating our dataset. These novel findings in the Portuguese population warrant further replication in additional independent studies, and provide additional candidates to more comprehensively understand IA etiopathogenesis.FCT grant: (CMUP-ERI/TPE/0028/2013), FCT fellowships and research contracts (SFRH/BPD/35737/2007, SFRH/BPD/70008/2010, SFRH/BD/43895/2008 and Ciência and Investigator- FCT contracts)

Bioengineered cell culture systems of central nervous system injury and disease

Cell culture systems, either 2D or explant based, have been pivotal to better understand the pathophysiology of several central nervous system (CNS) disorders. Recently, bioengineered cell culture systems have been proposed as an alternative to the traditional setups. These innovative systems often combine different cell populations in 3D environments that more closely recapitulate the different niches that exist within the developing or adult CNS. Given the importance of such systems for the future of CNS-related research, we discuss here the most recent advances in the field, particularly those dealing with neurodegeneration, neurodevelopmental disorders, and trauma.Financial support is acknowledged from Prémios Santa Casa Neurociências – Prize Melo e Castro for Spinal Cord Injury Research; Portuguese Foundation for Science and Technology [Doctoral fellowship (SFRH/BD/103075/2014) to E.D.G.; IF Development Grant to A.J.S.; Starting Grant to F. Marques; PostDoctoral fellowship SFRH/BPD/97701/2013 to N.A.S.]; this work was co-funded by Programa Operacional Regional do Norte (ON.2 – O Novo Norte), ao abrigo do Quadro de Referência Estratégico Nacional (QREN), através do Fundo Europeu de Desenvolvimento Regional (FEDER).info:eu-repo/semantics/publishedVersio

Inhibition of ovine in vitro fertilization by anti-Prt antibody: hypothetical model for Prt/ZP interaction

BACKGROUND: The impact of prion proteins in the rules that dictate biological reproduction is still poorly understood. Likewise, the role of prnt gene, encoding the prion-like protein testis specific (Prt), in ram reproductive physiology remains largely unknown. In this study, we assessed the effect of Prt in ovine fertilization by using an anti-Prt antibody (APPA) in fertilization medium incubated with spermatozoa and oocytes. Moreover, a computational model was constructed to infer how the results obtained could be related to a hypothetical role for Prt in sperm-zona pellucida (ZP) binding. METHODS: Mature ovine oocytes were transferred to fertilization medium alone (control) or supplemented with APPA, or pre-immune serum (CSerum). Oocytes were inseminated with ovine spermatozoa and after 18 h, presumptive zygotes (n = 142) were fixed to evaluate fertilization rates or transferred (n = 374) for embryo culture until D6-7. Predicted ovine Prt tertiary structure was compared with data obtained by circular dichroism spectroscopy (CD) and a protein-protein computational docking model was estimated for a hypothetical Prt/ZP interaction. RESULTS: The fertilizing rate was lower (P = 0.006) in APPA group (46.0+/−6.79%) when compared to control (78.5+/−7.47%) and CSerum (64.5+/−6.65%) groups. In addition, the cleavage rate was higher (P < 0.0001) in control (44.1+/−4.15%) than in APPA group (19.7+/−4.22%). Prt CD spectroscopy showed a 22% alpha-helical structure in 30% (m/v) aqueous trifluoroethanol (TFE) and 17% alpha in 0.6% (m/v) TFE. The predominant alpha-helical secondary structure detected correlates with the predicted three dimensional structure for ovine Prt, which was subsequently used to test Prt/ZP docking. Computational analyses predicted a favorable Prt-binding activity towards ZP domains. CONCLUSIONS: Our data indicates that the presence of APPA reduces the number of fertilized oocytes and of cleaved embryos. Moreover, the CD analysis data reinforces the predicted ovine Prt trend towards an alpha-helical structure. Predicted protein-protein docking suggests a possible interaction between Prt and ZP, thus supporting an important role for Prt in ovine fertilization

Evaluation of the antioxidant activity of extracts obtained form cherry seeds.

Annual cherry production in Portugal is around 19,000 tonnes, in an area of about 6,450 ha and covering about 11,100 farms, concentrated in some northern and central interior territories. It is also in these regions that in recent decades there has been a significant increase in farms specialized in the production of cherry, using new cultivars and new technologies in a business production model. Apart from being consumed in fresh form, cherries are used for many food preparations, like sweets, jellies or confectionary. In the plants that transform cherries, a significant amount of cherry seeds (also called cherry pits) is generated as residue or waste. The possible usage of these residues as raw material for extraction of compounds with antioxidant properties is beneficial in term of economic value as well as environmental impact. Hence, the objective of this work was to obtain extract rich in compounds with antioxidant activity from cherry seeds. The cherry seeds were obtained from a local waste management company, Nutrofertil, located in Tondela, in the district of Viseu (Portugal). They were grinded and then submitted to extraction procedures testing different operating conditions: magnetic stirrer versus ultrasound, different solvents (methanol, ethanol, water) and temperatures (from 35 ºC to 80 ºC). For the obtained extracts antioxidant activity was evaluated through spectrophotometric methods, using the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid)) radicals, and also the Ferric Reducing Antioxidant Power Assay (FRAP). All measurements were replicated at least trice, and were expressed as mg Trolox equivalents per gram (mg TE/g). The results obtained for the different experimental conditions indicated that least efficient extractions at ambient temperature were obtained with methanol using magnetic stirrer and with water using ultrasounds, for which the antioxidant activities measured by the DPPH method were 0.26 and 0.33 mg TE/g and by the ABTS method were 0.82 and 0.86 mg TE/g, respectively. Most efficient methods were water:ethanol (at 50% concentration) and water (100%), using magnetic stirrer in both cases. Highest antioxidant activity was obtained for water:ethanol by the DPPH method (0.72 mg TE/g) and for water (100%) by the ABTS method (1.25 mg TE/g). Tests with different concentrations for the aqueous solutions of ethanol and at different temperatures revealed that with increasing concentration of water the antioxidant diminished, from 0.62 to 0.27 mg TE/g at 35 ºC using the DPPH method. Additionally, the variation in temperature allowed reaching a maximum extraction of compounds with antioxidant activity at 70 ºC and decreasing thereafter. The maximum values obtained were registered at 70 ºC for all cases and were 0.74 mg TE/g for the water:ethanol 50:50 (v/v) by the DPPH method, 2.16 mg TE/g for the water:ethanol 60:40 (v/v) by the ABTS method and 3.43 mg TE/g for the water:ethanol 60:40 (v/v) by the FRAP method. The results obtained by the different methods were concordant in terms of the observed trends but giving different values of the measured antioxidant activity, which is a common characteristic observed in these types of evaluation techniques. This research allowed establishing some operational conditions that should be selected in order to maximize the extraction of compounds with antioxidant activity from cherry seeds. The use of ultrasounds was not found beneficial and the magnetic stirrer technique revealed to be more useful. Also the use of methanol was not found suitable, which is a good point given that this solvent is more pollutant and has more problems of toxicity. With respect to temperature, it was found that temperatures higher than 70 ºC are not beneficial because they induce the degradation of some bioactive compounds thus reducing the antioxidant activity of thee extracts.info:eu-repo/semantics/publishedVersio