102 research outputs found

    Proangiogenic Effect of 2A-Peptide Based Multicistronic Recombinant Constructs Encoding VEGF and FGF2 Growth Factors

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    Coronary artery disease remains one of the primary healthcare problems due to the high cost of treatment, increased number of patients, poor clinical outcomes, and lack of effective therapy. Though pharmacological and surgical treatments positively affect symptoms and arrest the disease progression, they generally exhibit a limited effect on the disease outcome. The development of alternative therapeutic approaches towards ischemic disease treatment, especially of decompensated forms, is therefore relevant. Therapeutic angiogenesis, stimulated by various cytokines, chemokines, and growth factors, provides the possibility of restoring functional blood flow in ischemic tissues, thereby ensuring the regeneration of the damaged area. In the current study, based on the clinically approved plasmid vector pVax1, multigenic constructs were developed encoding vascular endothelial growth factor (VEGF), fibroblast growth factors (FGF2), and the DsRed fluorescent protein, integrated via picornaviruses’ furin-2A peptide sequences. In vitro experiments demonstrated that genetically modified cells with engineered plasmid constructs expressed the target proteins. Overexpression of VEGF and FGF2 resulted in increased levels of the recombinant proteins. Concomitantly, these did not lead to a significant shift in the general secretory profile of modified HEK293T cells. Simultaneously, the secretome of genetically modified cells showed significant stimulating effects on the formation of capillary-like structures by HUVEC (endothelial cells) in vitro. Our results revealed that when the multicistronic multigene vectors encoding 2A peptide sequences are created, transient transgene co-expression is ensured. The results obtained indicated the mutual synergistic effects of the growth factors VEGF and FGF2 on the proliferation of endothelial cells in vitro. Thus, recombinant multicistronic multigenic constructs might serve as a promising approach for establishing safe and effective systems to treat ischemic diseases

    Chromium(III) biosorption onto spent grains residual from brewing industry : equilibrium, kinetics and column studies

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    The use of industrial wastes for wastewater treatment as a strategy to their re-use and valorisation may provide important advances toward sustainability. The present work gives new insights into heavy metal biosorption onto low-cost biosorbents, studying chromium(III) biosorption onto spent grains residual from a Portuguese brewing industry both in batch and expanded bed column systems. Experimental studies involved unmodified spent grains and spent grains treated with NaOH. Metal uptake followed a rapid initial step, well described by the pseudo-second-order kinetic model up to 27 h, indicating chemisorption to be the rate-limiting step. Beyond this period intraparticle diffusion assumed an important role in the uptake global kinetics. The best fit for equilibrium data was obtained using the Langmuir model, with unmodified spent grains having the higher maximum uptake capacity (q max = 16.7 mg g1). In open system studies, using expanded bed columns, the best performance was also achieved with unmodified spent grains: Breakthrough time (C/C i = 0.25) and total saturation time (C/C i = 0.99) occurred after 58 and 199 h of operation, corresponding to the accumulation of 390 mg of chromium(III), 43.3 % of the total amount entering the column. These results suggest that alkali treatment does not improve spent grains uptake performance. Changes in biomass composition determined by Fourier transform infrared spectroscopy suggested hydroxyl groups and proteins to have an important role in chromium(III) biosorption. This study points out that unmodified spent grains can be successfully used as low-cost biosorbent for trivalent chromium.The authors would like to thank the Portuguese brewing industry UNICER for all the support and FCT (Fundacao para a Ciencia e a Tecnologia) financial support through the Grant PRAXIS XXI/BD/15945/98

    Sustained Na<sup>+</sup>/H<sup>+</sup> exchanger activation promotes gliotransmitter release from reactive hippocampal astrocytes following oxygen-glucose deprivation

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    Hypoxia ischemia (HI)-related brain injury is the major cause of long-term morbidity in neonates. One characteristic hallmark of neonatal HI is the development of reactive astrogliosis in the hippocampus. However, the impact of reactive astrogliosis in hippocampal damage after neonatal HI is not fully understood. In the current study, we investigated the role of Na +/H+ exchanger isoform 1 (NHE1) protein in mouse reactive hippocampal astrocyte function in an in vitro ischemia model (oxygen/glucose deprivation and reoxygenation, OGD/REOX). 2 h OGD significantly increased NHE1 protein expression and NHE1-mediated H+ efflux in hippocampal astrocytes. NHE1 activity remained stimulated during 1-5 h REOX and returned to the basal level at 24 h REOX. NHE1 activation in hippocampal astrocytes resulted in intracellular Na+ and Ca2+ overload. The latter was mediated by reversal of Na+/Ca2+ exchange. Hippocampal astrocytes also exhibited a robust release of gliotransmitters (glutamate and pro-inflammatory cytokines IL-6 and TNFα) during 1-24 h REOX. Interestingly, inhibition of NHE1 activity with its potent inhibitor HOE 642 not only reduced Na+ overload but also gliotransmitter release from hippocampal astrocytes. The noncompetitive excitatory amino acid transporter inhibitor TBOA showed a similar effect on blocking the glutamate release. Taken together, we concluded that NHE1 plays an essential role in maintaining H + homeostasis in hippocampal astrocytes. Over-stimulation of NHE1 activity following in vitro ischemia disrupts Na+ and Ca2+ homeostasis, which reduces Na+-dependent glutamate uptake and promotes release of glutamate and cytokines from reactive astrocytes. Therefore, blocking sustained NHE1 activation in reactive astrocytes may provide neuroprotection following HI. © 2014 Cengiz et al

    The Influence of Hydrocarbon-Oxidizing Auxin-Producing Bacteria on the Growth, Biochemical Parameters, and Hormonal Status of Barley Plants in the Process of Bioremediation of Oil-Contaminated Soil

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    Взаимосвязи бактерий и растений в процессе биоремедиации почв, загрязненных нефтью, уделяется много внимания, однако воздействие бактерий-деструкторов нефти, синтезирующих фитогормоны, на содержание и распределение этих соединений в самих растениях, исследовано слабо. Целью полевого опыта было изучение влияния углеводородокисляющих бактерий, продуцирующих ауксины, на ростовые, биохимические показатели и гормональный статус растений ячменя в присутствии нефти и перспективы применения их ассоциаций для очистки почвы, содержащей нефть (в среднем 2,7 %). Обработка растений штаммами Enterobacter sp. UOM 3 и Pseudomonas hunanensis IB C7 приводила к увеличению длины и массы корней и побегов, индекса листовой поверхности и улучшению показателей элементов структуры урожая, которые были угнетены под воздействием поллютанта. В результате бактеризации повышалось содержание хлорофилла, флавоноидов и снижалось количество пролина. Наиболее заметным проявлением влияния бактерий на гормональную систему растений было уменьшение накопления абсцизовой кислоты. Полученные данные свидетельствуют о том, что интродукция микроорганизмов ослабляла для растений негативные последствия абиотического стресса, вызванного присутствием нефти. Совместное применение бактерий-нефтедеструкторов и растений эффективнее снижало содержание углеводородов в почве и увеличивало ее микробиологическую активность по сравнению с использованием их по отдельности. Изученные микробно-растительные комплексы признаны перспективными для биоремедиации нефтезагрязненных почвExtensive research has been done to investigate the relationship between bacteria and plants in the process of bioremediation of soils contaminated with oil, but the effect of oil-degrading bacteria that synthesize phytohormones on the content and distribution of these compounds in plants has been poorly studied. The aim of the field experiment was to study the effect of hydrocarbon-oxidizing bacteria producing auxins on the growth, biochemical parameters, and hormonal status of barley plants in the presence of oil and the prospects for using bacterial-plant associations for treating soil that contains oil (2.7 %, on average). Treatment of plants with cultures of Enterobacter sp. UOM 3 and Pseudomonas hunanensis IB C7 led to an increase in the length and mass of roots and shoots and the leaf surface index and an improvement in the parameters of the components of the crop structure that were suppressed by the pollutant. As a result of bacterization, the contents of chlorophyll and flavonoids increased, and the amount of proline decreased. The most noticeable effect of bacteria on the hormonal system of plants was a decrease in the accumulation of abscisic acid. The data obtained indicate that the treatment of plants with bacterial cultures alleviated the negative consequences of abiotic stress caused by the presence of oil for plants. The use of oil-degrading bacteria and plants in combination rather than separately more effectively reduced the content of hydrocarbons in the soil and increased its microbiological activity. The microbial-plant combinations studied in this work are regarded as promising for the bioremediation of oil-contaminated soil

    Monitoring biological wastewater treatment processes: Recent advances in spectroscopy applications

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    Biological processes based on aerobic and anaerobic technologies have been continuously developed to wastewater treatment and are currently routinely employed to reduce the contaminants discharge levels in the environment. However, most methodologies commonly applied for monitoring key parameters are labor intensive, time-consuming and just provide a snapshot of the process. Thus, spectroscopy applications in biological processes are, nowadays, considered a rapid and effective alternative technology for real-time monitoring though still lacking implementation in full-scale plants. In this review, the application of spectroscopic techniques to aerobic and anaerobic systems is addressed focusing on UV--Vis, infrared, and fluorescence spectroscopy. Furthermore, chemometric techniques, valuable tools to extract the relevant data, are also referred. To that effect, a detailed analysis is performed for aerobic and anaerobic systems to summarize the findings that have been obtained since 2000. Future prospects for the application of spectroscopic techniques in biological wastewater treatment processes are further discussed.The authors thank the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, COMPETE 2020 (POCI-01-0145-FEDER-006684) and the project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. The authors also acknowledge the financial support to Daniela P. Mesquita and Cristina Quintelas through the postdoctoral Grants (SFRH/BPD/82558/2011 and SFRH/BPD/101338/2014) provided by FCT - Portugal.info:eu-repo/semantics/publishedVersio

    Measuring collective action intention toward gender equality across cultures

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    Collective action is a powerful tool for social change and is fundamental to women and girls’ empowerment on a societal level. Collective action towards gender equality could be understood as intentional and conscious civic behaviors focused on social transformation, questioning power relations, and promoting gender equality through collective efforts. Various instruments to measure collective action intentions have been developed, but to our knowledge none of the published measures were subject to invariance testing. We introduce the gender equality collective action intention (GECAI) scale and examine its psychometric isomorphism and measurement invariance, using data from 60 countries (N = 31,686). Our findings indicate that partial scalar measurement invariance of the GECAI scale permits conditional comparisons of latent mean GECAI scores across countries. Moreover, this metric psychometric isomorphism of the GECAI means we can interpret scores at the country-level (i.e., as a group attribute) conceptually similar to individual attributes. Therefore, our findings add to the growing body of literature on gender based collective action by introducing a methodologically sound tool to measure collective action intentions towards gender equality across cultures

    Measuring collective action intention toward gender equality across cultures

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    Collective action is a powerful tool for social change and is fundamental to women and girls’ empowerment on a societal level. Collective action towards gender equality could be understood as intentional and conscious civic behaviors focused on social transformation, questioning power relations, and promoting gender equality through collective efforts. Various instruments to measure collective action intentions have been developed, but to our knowledge none of the published measures were subject to invariance testing. We introduce the gender equality collective action intention (GECAI) scale and examine its psychometric isomorphism and measurement invariance, using data from 60 countries (N = 31,686). Our findings indicate that partial scalar measurement invariance of the GECAI scale permits conditional comparisons of latent mean GECAI scores across countries. Moreover, this metric psychometric isomorphism of the GECAI means we can interpret scores at the country-level (i.e., as a group attribute) conceptually similar to individual attributes. Therefore, our findings add to the growing body of literature on gender based collective action by introducing a methodologically sound tool to measure collective action intentions towards gender equality across cultures.info:eu-repo/semantics/acceptedVersio

    The Psychological Science Accelerator's COVID-19 rapid-response dataset

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