Contribution of PRS3, RPB4 and ZWF1 to the resistance of industrial Saccharomyces cerevisiae CCUG53310 and PE-2 strains to lignocellulosic hydrolysate-derived inhibitors

PRS3, RPB4 and ZWF1 were previously identified as key genes for yeast tolerance to lignocellulose-derived inhibitors. To better understand their contribution to yeast resistance to the multiple stresses occurring during lignocellulosic hydrolysate fermentations, we overexpressed these genes in two industrial Saccharomyces cerevisiae strains, CCUG53310 and PE-2, and evaluated their impact on the fermentation of Eucalyptus globulus wood and corn cob hydrolysates. PRS3 overexpression improved the fermentation rate (up to 32%) and productivity (up to 48%) in different hydrolysates. ZWF1 and RPB4 overexpression did not improve the fermentation performance, but their increased expression in the presence of acetic acid, furfural and hydroxymethylfurfural was found to contribute to yeast adaptation to these inhibitors. This study expands our understanding about the molecular mechanisms involved in industrial yeast tolerance to the stresses occurring during lignocellulosic bioethanol production and highlights the importance of selecting appropriate strain backgrounds/hydrolysates combinations when addressing further improvement of these processes.Fundação para a Ciência e a Tecnologia, Portugal, through: Grant SFRH/BDP/63831/2009 to Grant SFRH/BPD/77995/2011; Strategic Project PEst-OE/EQB/LA0023/2013; and Project BioInd NORTE-07-0124-FEDER-000028 co-funded by the Programa Operacional Regional do Norte (ON.2 – O Novo Norte), QREN, FEDER

Advances in isoflavone profile characterisation using matrix solid-phase dispersion coupled to HPLC/DAD in medicago species

Analytical methods used in phytochemistry analysis are limited by the sample preparation step, which should ideally be fast, accurate, ecofriendly and achievable using low quantities of the sample. Matrix solid-phase dispersion (MSPD) may be a good alternative for combining extraction and purification procedures, thereby reducing the indicated limitations. Objective-Applying an MSPD extraction procedure coupled to high-performance liquid chromatography diode-array detection (HPLC/DAD) as an alternative methodology to evaluate isoflavone profiles. Methods-Isoflavone profiles were determined for the leaves of nine species of Medicago in the late flower phenological stage (one or more nodes with 50% open flowers, no seed pods). Extraction was performed following MSPD, and isoflavone profiles were characterised using HPLC/DAD. The quantified amounts were compared with previous results in different species commonly recognised as good sources of isoflavones. Results-Formononetin was the major isoflavone in most species, except M. polymorpha and M. truncatula. The isoflavone amounts were significantly different among the assayed species, with M. orbicularis and M. arabica as the major isoflavone sources, while M. rigidula presented the lowest contents. Furthermore, the detected differences allow electing the best species as a primary source of a specific isoflavone. Conclusion-The MSPD allowed good extraction efficiency, reproducibility and recovery. Some of the species showed relevant isoflavone contents, even when compared with acknowledged plant sources such as soy or red clover. To the best of our knowledge the results presented are reported for the first time in these species.T. Visnevschi-Necrasov and João C.M. Barreira are grateful to ‘FCT - Fundação para a Ciência e a Tecnologia’ for their grants (BD/ 31734/2006 and SFRH/BPD/72802/2010, respectively). S.C. Cunha is grateful to ‘POPH-QREN - Tipologia 4.2, Fundo Social Europeu e Fundo Nacional MCTES’.info:eu-repo/semantics/publishedVersio

Phylogenetic insights on the isoflavone profile variations in Fabaceae spp.: Assessment through PCA and LDA

Legumes (Fabaceae) are important crops, known as sources of food, feed for livestock and raw materials for industry. Their ability to capture atmospheric nitrogen during symbiotic processes with soil bacteria reduces the need for expensive chemical fertilizers, improving soil and water quality. Several Fabaceae species are acknowledged for the high levels of secondary metabolites. Isoflavones are among the most well-known examples of these compounds, being recognized for their several types of biological activity. Herein, isoflavone profiles were characterized in nine species of four Fabaceae genera (Biserrula, Lotus, Ornithopus and Scorpiurus). Plants were harvested in the late flower physiological stage to prevent biased results due to naturally occurring variations along the vegetative cycle. Isoflavones were extracted using matrix solid-phase dispersion and analyzed by high performance liquid chromatography/diode-array detection. The detected profiles revealed significant differences, inclusively among species belonging to the same genus, indicating that other factors besides the genotypic features contribute to the expression of these phenolic compounds. The classification of the results by principal component analysis placed species belonging to the same genus in different clustering groups, proving this latter assumption. However, the detected profiles proved to be characteristic of the assayed, as it was proved by the applied linear discriminant analysis.info:eu-repo/semantics/publishedVersio

Medicago spp. as potential sources of bioactive isoflavones: Characterization according to phylogenetic and phenologic factors

A high variety of plant species are often proposed as potential natural sources of specific bioactive components, with emphasis in phenolic compounds. However, the ability to produce a determined phytochemical might be variable, even among species with close phylogeny. Furthermore, the metabolic dynamics vary greatly according to phenologic factors. Herein, it was verified whether isoflavone production in Medicago spp. is more associated with phylogenetic or phenologic determinants, to define the optimal productive conditions. Isoflavone profiles were characterized in field-grown Medicago species in three phenologic stages. Isoflavones were extracted by matrix solid-phase dispersion method and analyzed using high-performance liquid chromatography coupled with a diode-array detector. The obtained data were evaluated by a generalized linear model (GLM) and linear discriminant analysis (LDA). Formononetin, genistein and irilone were the most abundant isoflavones, reaching values higher than those present in acknowledged plant sources like soy or red clover. Outputs from GLM and LDA indicate that the phylogenetic factors are the most defining criteria. This study promotes Medicago spp. as potential isoflavone sources, particularly because the effects of these compounds are highly dependent on their type and concentration, with potential application as foodstuff, feedstuff, or in the nutraceutical and pharmaceutica l industry.Visnevschi-Necrasov and João C.M. Barreira are grateful to ‘‘FCT – Fundação para a Ciência e a Tecnologia’’ for their Grants (SFRH/BD/31734/2006 and SFRH/BPD/72802/2010, respectively). S.C. Cunha is grateful to ‘‘POPH-QREN – Tipologia 4.2, Fundo Social Europeu e Fundo Nacional MCTES’’.info:eu-repo/semantics/publishedVersio

Efeito da Reeducação Postural Global sobre o sistema cardiovascular de indivíduos saudáveis

ObjectiveTo evaluate the effect of Global Postural Reeducation (GPR) on cardiovascular system by heart rate variability (HRV), blood pressure (BP) and heart rate (HR). Materials and methodSeventeen healthy men (22.47 ± 3.02 years) were submitted to the postures frog on the floor, frog on the air, sitting, standing against the wall and inclined standing, two postures per session. The systolic and diastolic blood pressure (SBP and DBP) and HR were recorded. The intervals between heartbeats were collected during the whole session (Polar S810i). The frequency domain was analyzed (Wavelet Transform), the low frequency (LF) and high frequency (HF) were obtained. The data were analyzed by ANOVA and Tukey (p < 0.05). ResultIncreased LF/HF ratio was observed in the frog on the floor (1 ± 0.1 vs. 2 ± 0.3 p < 0.05) and on the air postures (1 ± 0.1 vs. 2 ± 0.2 p < 0.01). There was an increase in SBP in the postures frog on the floor (123 ± 2 vs. 136 ± 4 p < 0.05), frog on the air (122 ± 2 vs. 133 ± 3 p < 0.05), standing against the wall (123 ± 2 vs. 136 ± 4 p < 0.05), inclined standing (124 ± 3 vs. 146 ± 5 p < 0.05). There was increase of DBP in the postures frog on the floor (69 ± 2 vs. 81 ± 2 p < 0.01), frog on the air (72 ± 2 vs. 83 ± 3 p < 0.05), sitting (85 ± 2 vs. 102 ± 3 p < 0.01). There was increase in HR in the postures frog on the air (67 ± 2 vs. 77 ± 3 p < 0.05) and inclined standing (88 ± 3.5 vs. 101 ± 3 p < 0.05). ConclusionThe increase in LF/HF ratio and also the BP and HR indicates high sympathetic activity, possibly related to the work isometric developed during GPR method. [P]ObjetivoAvaliar o efeito da Reeducação Postural Global (RPG) sobre o sistema cardiovascular por meio da variabilidade da frequência cardíaca (VFC), pressão arterial (PA) e frequência cardíaca (FC). Materiais e métodoDezessete homens saudáveis (22,47± 3,02 anos) foram submetidos às posturas rã no chão, rã no ar, sentada, de pé contra a parede e de pé inclinada, sendo duas posturas por sessão. A pressão arterial sistólica e diastólica (PAS, PAD) e FC foram registradas. Os intervalos entre os batimentos cardíacos foram coletados durante toda a sessão (Polar S810i). O domínio da frequência foi analisado (transformada Wavelet), as bandas de baixa (LF) e a alta frequências (HF) foram obtidas. Os dados foram analisados por ANOVA e Tukey (p < 0,05). ResultadoHouve aumento da relação LF/HF nas posturas rã no chão (1 ± 0,1 vs. 2 ± 0,3 p < 0,05); rã no ar (1 ± 0,1 vs. 2 ± 0,2 p < 0,01). Observou-se aumento da PAS nas posturas rã no chão (123 ± 2 vs. 136 ± 4 p < 0,05), rã no ar (122 ± 2 vs. 133 ± 3 p < 0,05), de pé na parede (123 ± 2 vs. 136 ± 4 p < 0,05), de pé inclinada (124 ± 3 vs. 146 ± 5 p < 0,05). Houve aumento da PAD nas posturas rã no chão (69 ± 2 vs. 81 ± 2 p < 0,01), rã no ar (72 ± 2 vs. 83 ± 3 p < 0,05), sentada (85 ± 2 vs. 102 ± 3 p < 0,01). Houve aumento da FC nas posturas rã no ar (67 ± 2 vs. 77 ± 3 p < 0,05) e de pé inclinada (88 ± 3,5 vs. 101 ± 3 p < 0,05). ConclusãoO aumento da relação LF/HF e também da PA e da FC aponta para maior atividade simpática, possivelmente relacionada ao trabalho isométrico desenvolvido durante a realização do método de RPG.Universidade PaulistaUniversidade Federal de São Paulo (UNIFESP)Universidade do Vale do ParaíbaUNIFESPSciEL

A new brain emotional learning simulink toolbox for control systems design

The brain emotional learning (BEL) control paradigm has been gathering increased interest by the control systems design community. However, the lack of a consistent mathemat- ical formulation and computer based tools are factors that have prevented its more widespread use. In this article both features are tackled by providing a coherent mathematical framework for both the continuous and discrete-time formulations and by presenting a Simulink R computational tool that can be easily used for fast prototyping BEL based control systems.info:eu-repo/semantics/publishedVersio

Mitochondrial apoptosis-inducing factor is involved in doxorubicin-induced toxicity on H9c2 cardiomyoblasts

AbstractThe cardiotoxicity induced by the anti-cancer doxorubicin involves increased oxidative stress, disruption of calcium homeostasis and activation of cardiomyocyte death. Nevertheless, antioxidants and caspase inhibitors often show little efficacy in preventing cell death. We hypothesize that a caspase-independent cell death mechanism with the release of the apoptosis-inducing factor from mitochondria is involved in doxorubicin toxicity. To test the hypothesis, H9c2 cardiomyoblasts were used as model for cardiac cells. Our results demonstrate that z-VAD-fmk, a pan-caspase inhibitor, does not prevent doxorubicin toxicity in this cell line. Doxorubicin treatment results in AIF translocation to the nuclei, as confirmed by Western Blotting of cell fractions and confocal microscopy. Also, doxorubicin treatment of H9c2 cardiomyoblasts resulted in the appearance of 50kbp DNA fragments, a hallmark of apoptosis-inducing factor nuclear effects. Apoptosis-inducing factor knockdown using a small-interfering RNA approach in H9c2 cells resulted in a reduction of doxorubicin toxicity, including decreased p53 activation and poly-ADP-ribose-polymerase cleavage. Among the proteases that could be responsible for apoptosis-inducing factor cleavage, doxorubicin decreased calpain activity but increased cathepsin B activation, with inhibition of the latter partly decreasing doxorubicin toxicity. Altogether, the results support that apoptosis-inducing factor release is involved in doxorubicin-induced H9c2 cell death, which explains the limited ability of caspase inhibitors to prevent toxicity

Histone Deacetylase Inhibitors Protect Against Pyruvate Dehydrogenase Dysfunction in Huntington's Disease

Transcriptional deregulation and changes in mitochondrial bioenergetics, including pyruvate dehydrogenase (PDH) dysfunction, have been described in Huntington's disease (HD). We showed previously that the histone deacetylase inhibitors (HDACIs) trichostatin A and sodium butyrate (SB) ameliorate mitochondrial function in cells expressing mutant huntingtin. In this work, we investigated the effect of HDACIs on the regulation of PDH activity in striatal cells derived from HD knock-in mice and YAC128 mice. Mutant cells exhibited decreased PDH activity and increased PDH E1alpha phosphorylation/inactivation, accompanied by enhanced protein levels of PDH kinases 1 and 3 (PDK1 and PDK3). Exposure to dichloroacetate, an inhibitor of PDKs, increased mitochondrial respiration and decreased production of reactive oxygen species in mutant cells, emphasizing PDH as an interesting therapeutic target in HD. Treatment with SB and sodium phenylbutyrate, another HDACI, recovered cell viability and overall mitochondrial metabolism in mutant cells. Exposure to SB also suppressed hypoxia-inducible factor-1 (HIF-1α) stabilization and decreased the transcription of the two most abundant PDK isoforms, PDK2 and PDK3, culminating in increased PDH activation in mutant cells. Concordantly, PDK3 knockdown improved mitochondrial function, emphasizing the role of PDK3 inactivation on the positive effects achieved by SB treatment. YAC128 mouse brain presented higher mRNA levels of PDK1-3 and PDH phosphorylation and decreased energy levels that were significantly ameliorated after SB treatment. Furthermore, enhanced motor learning and coordination were observed in SB-treated YAC128 mice. These results suggest that HDACIs, particularly SB, promote the activity of PDH in the HD brain, helping to counteract HD-related deficits in mitochondrial bioenergetics and motor function.SIGNIFICANCE STATEMENT The present work provides a better understanding of mitochondrial dysfunction in Huntington's disease (HD) by showing that the pyruvate dehydrogenase (PDH) complex is a promising therapeutic target. In particular, the histone deacetylase inhibitor sodium butyrate (SB) may indirectly (through reduced hypoxia-inducible factor 1 alpha stabilization) decrease the expression of the most abundant PDH kinase isoforms (e.g., PDK3), ameliorating PDH activity and mitochondrial metabolism and further affecting motor behavior in HD mice, thus constituting a promising agent for HD neuroprotective treatment.FCT, Santa Casa da Misericórdia de Lisboa (SCML), Fundação Luso-Americana para o Desenvolvimento (FLAD) Life Scienc

Alignment of legislation of Brazilian states and municipalities with a National Solid Waste Policy

The establishment of policies aimed at reducing production, disposal and adequate discarding of waste to reduce environmental impacts has been evidenced throughout the world in this century. In this study, the degree of alignment between federal and state and municipal legislations regarding solid waste in municipalities of the same size of Joinville, Santa Catarina was analyzed and the practices on solid waste plans, selective col-lection and final disposal were investigated. The selection of these was based on official sociodemographic data using as criteria: population, population density, human development index and belonging to different regions of Brazil. The confrontation of the state and municipal legislations with national law occurred by pre-established criteria. The results indicated a misalignment in the concepts between state and municipal legislations with federal law. The framework of the Joinville’s law was satisfactory. The review and update these legislation and practices involving the management of solid waste has been evidenced, especially in the municipalities of Juiz de Fora, Minas Gerais and Porto Velho, Rondônia, which need to be adjusted in relation to the dead-lines established by federal law