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The objective of this research was to evaluate the performance of a structured bed reactor (SBRIA), carried out with intermittent aeration (IA), in the removal of organic matter and nitrogen from dairy effluent, when run with different organic loading rates (OLR). The SBRIA was operated for 227 days, with 2:1 AI cycles (2 h with aeration on and 1 h off) and Hydraulic Retention Time (HRT) of 16 h. Three phases, with different OLR, were evaluated: phases A (1000 gCOD m-3 day-1 - 63 days), B (1400 gCOD m-3 day-1 - 94 days), and C (1800 gCOD m-3 day-1 - 70 days). The percentage of COD, NH4+-N removal, and nitrogen removal, respectively, were above 85 ± 7%, 73 ± 27%, and 83 ± 5, in all phases. There was no accumulation of the oxidized forms of nitrogen in the reactor. The kinetic test, performed to evaluate the nitrification and denitrification in the system, indicated that even in dissolved oxygen concentrations of 4.5 mg L-1, it was possible to obtain the denitrification process in the system. The results demonstrate that the reactor under study has positive characteristics to be used as an alternative for removing the removal of organic material and nitrogen in the biological treatment of dairy effluents.

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High concentrations of inorganic arsenic in groundwater for human consumption is a worldwide common problem. Particularly, the determination of As(III) becomes important, since this species is more toxic than organic, pentavalent and elemental arsenic forms. In this work, a 3D-printed device that included a 24-well microplate was developed to perform the colourimetric kinetic determination of arsenic (III) by digital movie analysis. A smartphone camera attached to the device was used to take the movie during the process where As(III) inhibited the decolourization of methyl orange. The movie images were subsequently transformed from RGB to YIQ space to obtain a new analytical parameter called "d", which was related to the chrominance of the image. Then, this parameter allowed the determination of the inhibition time of reaction (tin), which was linearly correlated with the concentration of As(III). A linear calibration curve (R = 0.9995) in the range from 5 µg L-1 to 200 µg L-1 was obtained. The method was precise (RSD = 1.2%), and the limits of detection (LOD) and quantification (LOQ) were 1.47 µg L-1 and 4.44 µg L-1, respectively. These values were lower than the limit established by the World Health Organization for total arsenic in drinking water (10 µg L-1). The accuracy of the method was assessed by a recovery study with optimal results (94.3%-104.0%). Additionally, the Analytical GREEnness metric approach was applied, obtaining a score 1.7 times higher than previously published works. The method is simple, portable and low-cost, being in compliance with various principles of green analytical chemistry.

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Prior to clinical use, extensive in vitro proliferation of human adipose-derived stem cells (ASCs) is required. Among the current options, spinner-type stirred flasks, which use microcarriers to increase the yield of adherent cells, are recommended. Here, we propose a methodology for ASCs proliferation through cell suspension culture using Cultispher-S® microcarriers (MC) under agitation in a spinner flask, with the aim of establishing a system that reconciles the efficiency of cell yield with high viability of the culture during two distinct phases: seeding and proliferation. The results showed that cell adhesion was potentiated under intermittent stirring at 70 rpm in the presence of 10% FBS for an initial cell concentration of 2.4 × 104 cells/mL in the initial 24 h of cultivation. In the proliferation phase, kinetic analysis showed that cell growth was higher under continuous agitation at 50 rpm with a culture medium renewal regime of 50% every 72 h, which was sufficient to maintain the culture at optimal levels of nutrients and metabolites for up to nine days of cultivation, representing an 11.1-fold increase and a maximum cell productivity of 422 cells/mL/h (1.0 × 105 viable cells/mL). ASCs maintained the immunophenotypic characteristics and mesodermal differentiation potential of both cell lines from different donors. The established protocol represents a more efficient and cost-effective method to obtain a high proliferation rate of ASCs in a microcarrier-based system, which is necessary for large-scale use in cell therapy, highlighting that the manipulation of critical parameters optimizes the ASCs production process.

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The pyrolysis process is a thermochemical recycling process that in recent years has gained importance due to its application in plastic waste, which is one of the biggest environmental problems today. Thus, it is essential to carry out kinetic and thermodynamic analyses to understand the thermocatalytic degradation processes involved in plastic waste mixtures. In this sense, the main objective of this study is to analyze the degradation kinetics of the specific mixture of polypropylene (25%) and polystyrene (75%) with 10% mass of regenerated FCC catalyst which was recovered from conventional refining processes using 3 heating rates at 5, 10 and 15 K min-1 by thermogravimetric analysis (TGA). The obtained TGA data were compared with the isoconversional models used in this work that include Friedman (FR), Kissinger Akahira Sunose (KAS), Flynn-Wall-Ozawa (FWO), Starink (ST) and Miura-Maki (MM) in order to determine the one that best fits the experimental data and to analyze the activation energy and the pre-exponential factor; the model is optimized by means of the difference of minimum squares. Activation energy values between 148 and 308 kJ/mol were obtained where the catalytic action has been notorious, decreasing the activation energy values with respect to thermal processes.

RESUMO

Our study demonstrated the energy gains when using biomass from three macrophyte, used commonly in constructed wetlands for wastewater treatment, the water hyacinth, cattail, and dwarf papyrus, as a substrate for biogas generation. The biochemical methane potential for the three biomass was evaluated in batch and at bench at 37 °C. A kinetic analysis of anaerobic digestion was also conducted for these substrates, evaluating the biogas composition and energy potential. Anaerobic digestion resulted in 94.27, and 25 mLCH4/gVSsubstrate of dry mass; and 19,569.65, 5617.88, and 6068.45 kJ/t of cattail, water hyacinth, and dwarf papyrus, respectively. Biomass from water hyacinth did sustain the fastest degradation, indicating that models considering the lag phase are more adequate to evaluate the anaerobic digestion of this type of substrate. Higher digestion speed resulted in the generation of 2901.88 kJ/t more energy with biomass from water hyacinth versus cattail, highlighting its value for use in constructed wetlands.

RESUMO

Prior to clinical use, extensive in vitro proliferation of human adipose-derived stem cells (ASCs) is required. Among the current options, spinner-type stirred flasks, which use microcarriers to increase the yield of adherent cells, are recommended. Here, we propose a methodology for ASCs proliferation through cell suspension culture using Cultispher-S® microcarriers (MC) under agitation in a spinner flask, with the aim of establishing a system that reconciles the efficiency of cell yield with high viability of the culture during two distinct phases: seeding and proliferation. The results showed that cell adhesion was potentiated under intermittent stirring at 70 rpm in the presence of 10% FBS for an initial cell concentration of 2.4 × 104 cells/mL in the initial 24 h of cultivation. In the proliferation phase, kinetic analysis showed that cell growth was higher under continuous agitation at 50 rpm with a culture medium renewal regime of 50% every 72 h, which was sufficient to maintain the culture at optimal levels of nutrients and metabolites for up to nine days of cultivation, representing an 11.1-fold increase and a maximum cell productivity of 422 cells/mL/h (1.0 × 105 viable cells/mL). ASCs maintained the immunophenotypic characteristics and mesodermal differentiation potential of both cell lines from different donors. The established protocol represents a more efficient and cost-effective method to obtain a high proliferation rate of ASCs in a microcarrier-based system, which is necessary for large-scale use in cell therapy, highlighting that the manipulation of critical parameters optimizes the ASCs production process.

RESUMO

A challenge in the study of gastrointestinal microbiota (GITm) is the validation of the genomic data with metabolic studies of the microbial communities to understand how the microbial networks work during health and sickness. To gain insights into the metabolism of the GITm, feces from healthy and sick rats with cancer were inoculated in a defined synthetic medium directed for anaerobic prokaryote growth (INC-07 medium). Significant differences between cultures of healthy and sick individuals were found: 1) the consumption of the carbon source and the enzyme activity involved in their catabolism (e.g., sucrase, lactase, lipases, aminotransferases, and dehydrogenases); 2) higher excretion of acetic, propionic, isobutyric, butyric, valeric, and isovaleric acids; 3) methane production; 4) ability to form biofilms; and 5) up to 500 amplicon sequencing variants (ASVs) identified showed different diversity and abundance. Moreover, the bowel inflammation induced by cancer triggered oxidative stress, which correlated with deficient antioxidant machinery (e.g., NADPH-producing enzymes) determined in the GITm cultures from sick individuals in comparison with those from control individuals. Altogether, the data suggested that to preserve the microbial network between bacteria and methanogenic archaea, a complete oxidation of the carbon source may be essential for healthy microbiota. The correlation of 16S rRNA gene metabarcoding between cultures and feces, as well as metabolomic data found in cultures, suggest that INC-07 medium may be a useful tool to understand the metabolism of microbiota under gut conditions.

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International authorities classify the ricin toxin, present in castor seeds, as a potential agent for use in bioterrorism. Therefore, the detection, identification, and characterization of ricin are considered the first actions for its risk assessment during a suspected exposure, parallel to the development of therapeutic and medical countermeasures. In this study, we report the kinetic analysis of electro-oxidation of adenine released from hsDNA by the catalytic action of ricin by square wave voltammetry. The results suggest that ricin-mediated adenine release exhibited an unusual kinetic profile, with a progress curve controlled by the accumulation of the product and the values of the kinetic constants of 46.6 µM for Km and 2000 min-1 for kcat, leading to a catalytic efficiency of 7.1 × 105 s-1 M-1.

RESUMO

International authorities classify ricin toxin present in castor seed as a potential agent for use in bioterrorism. Therefore, the detection, identification, and characterization of ricin in various sample matrices are considered necessary actions for risk assessment during a suspected exposure. This study reports a portable electrochemical assay for detecting active ricin based on the adenine electro-oxidation released from herring sperm DNA substrate by its catalytic action. Also, kinetic parameters were calculated, and the values were Km of 3.14 µM and Kcat 2107 min-1. A linear response was found in optimized experimental conditions for ricin concentrations ranging from 8 to 120 ng/mL, and with a detection limit of 5.14 ng/mL. This proposed detection strategy emphasizes the possibility of field detection of active ricin in food matrices and can be applied to other endonucleolytic activities.

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The presence of hazardous jarosites causes a serious environmental problems, releasing potentially toxic elements, principally heavy metals such as Pb, As, Tl, Cr among others to the environment. Thus, the dissolution process of jarosites has to be monitored to assess the environmental impact. In the present work, the different hazardous jarosites were prepared, and characterized by analytical techniques (XRD, SEM, EDS, etc.), and the composition of jarosites was determined by induction-coupled plasma spectroscopy (ICP). Shrinking core kinetic model (SCKM) was employed to understand the stability of hazardous jarosites, studying a complete kinetic analysis of the jarosite dissolution process under different conditions (temperatures and pH). The results show that temperature has the highest effect on stability followed by pH, requiring extreme parameters for high dissolution. The batch experiments show that the results are in good agreement with the SCKM forming a solid layer as by-products. The chemical reaction, i.e. dissolution process performs through mostly controlling stage at extreme pH values and then moved to mass transport in the fluid layer. After analyzing the results, a kinetic equation has been proposed to describe adequately the dissolution process, and it predicts the lifetime of the hazardous jarosites.

RESUMO

Although membrane proteins constitute an important class of biomolecules involved in key cellular processes, study of the thermodynamic and kinetic stability of their structures is far behind that of soluble proteins. It is known that many membrane proteins become unstable when removed by detergent extraction from the lipid environment. In addition, most of them undergo irreversible denaturation, even under mild experimental conditions. This process was found to be associated with partial unfolding of the polypeptide chain exposing hydrophobic regions to water, and it was proposed that the formation of kinetically trapped conformations could be involved. In this review, we will describe some of the efforts toward understanding the irreversible inactivation of membrane proteins. Furthermore, its modulation by phospholipids, ligands, and temperature will be herein discussed.

RESUMO

This study determined the specific uptake rate of glucose and corn oil substrates used as carbon sources in batch cultures of Gibberella fujikuroi. We tested three biological models of growth rate: Monod, logistic and lag-exponential. With respect to the substrate consumption rate, we tested two models: constant cell yield (CCY) and law of mass action (LMA). The experimental data obtained from the culture with glucose as substrate correlated satisfactorily with the logistic/LMA model, indicating that the cell yield was variable. In the case of corn oil as carbon source, considering total residual lipids as substrate in the culture broth, the model with the best correlation was the lag-exp/CCY model. The quantification by GC of the three main fatty acids (linoleic, oleic and palmitic) in the culture medium showed a cumulative behavior, with a maximum concentration of each acid at 36 h. We established a more explicit mechanism of the consumption of corn oil, consisting of two stages: generation of fatty acids by hydrolysis and consumption by cellular uptake. The kinetic of hydrolysable lipids was of first order. We found that the hydrolysis rate of corn oil is not a limiting factor for the uptake of fatty acids by the microorganism. We also established, based on the analysis of the identical mathematical structure of consumption kinetics, that the uptake of fatty acids is faster than the uptake of glucose.

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Two different putative galactokinase genes, found in the genome database of Trypanosoma cruzi were cloned and sequenced. Expression of the genes in Escherichia coli resulted for TcGALK-1 in the synthesis of a soluble and active enzyme, and in the case of TcGALK-2 gene a less soluble protein, with predicted molecular masses of 51.9kDa and 51.3kDa, respectively. The Km values determined for the recombinant proteins were for galactose 0.108mM (TcGALK-1) and 0.091mM (TcGALK-2) and for ATP 0.36mM (TcGALK-1) and 0.1mM (TcGALK-2). Substrate inhibition by ATP (Ki 0.414mM) was only observed for TcGALK-2. Gel-filtration chromatography showed that natural TcGALKs and recombinant TcGALK-1 are monomeric. In agreement with the possession of a type-1 peroxisome-targeting signal by both TcGALKs, they were found to be present inside glycosomes using two different methods of subcellular fractionation in conjunction with mass spectrometry. Both genes are expressed in epimastigote and trypomastigote stages since the respective proteins were immunodetected by western blotting. The T. cruzi galactokinases present their highest (52-47%) sequence identity with their counterpart from Leishmania spp., followed by prokaryotic galactokinases such as those from E. coli and Lactococcus lactis (26-23%). In a phylogenetic analysis, the trypanosomatid galactokinases form a separate cluster, showing an affiliation with bacteria. Epimastigotes of T. cruzi can grow in glucose-depleted LIT-medium supplemented with 20mM of galactose, suggesting that this hexose, upon phosphorylation by a TcGALK, could be used in the synthesis of UDP-galactose and also as a possible carbon and energy source.

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Porous silicon (PSi) is a nanostructured material possessing a huge surface area per unit volume. In consequence, the adsorption and diffusion of oxygen in PSi are particularly important phenomena and frequently cause significant changes in its properties. In this paper, we study the thermal oxidation of p+-type free-standing PSi fabricated by anodic electrochemical etching. These free-standing samples were characterized by nitrogen adsorption, thermogravimetry, atomic force microscopy and powder X-ray diffraction. The results show a structural phase transition from crystalline silicon to a combination of cristobalite and quartz, passing through amorphous silicon and amorphous silicon-oxide structures, when the thermal oxidation temperature increases from 400 to 900 °C. Moreover, we observe some evidence of a sinterization at 400 °C and an optimal oxygen-absorption temperature about 700 °C. Finally, the UV/Visible spectrophotometry reveals a red and a blue shift of the optical transmittance spectra for samples with oxidation temperatures lower and higher than 700 °C, respectively.

RESUMO

The thermal behavior of two polymorphic forms of rifampicin was studied by DSC and TG/DTG. The thermoanalytical results clearly showed the differences between the two crystalline forms. Polymorph I was the most thermally stable form, the DSC curve showed no fusion for this species and the thermal decomposition process occurred around 245 ºC. The DSC curve of polymorph II showed two consecutive events, an endothermic event (Tpeak = 193.9 ºC) and one exothermic event (Tpeak = 209.4 ºC), due to a melting process followed by recrystallization, which was attributed to the conversion of form II to form I. Isothermal and non-isothermal thermogravimetric methods were used to determine the kinetic parameters of the thermal decomposition process. For non-isothermal experiments, the activation energy (Ea) was derived from the plot of Log β vs 1/T, yielding values for polymorph form I and II of 154 and 123 kJ mol-1, respectively. In the isothermal experiments, the Ea was obtained from the plot of lnt vs 1/T at a constant conversion level. The mean values found for form I and form II were 137 and 144 kJ mol-1, respectively.

O comportamento térmico de duas formas polimórficas da rifampicina foi estudado por DSC e TG/DTG. Os resultados termoanalíticos mostraram claramente as diferenças entre as duas formas cristalinas. O polimorfo I é a forma mais estável termicamente, a curva DSC não mostrou a fusão dessa espécie e o processo de decomposição térmica ocorreu próximo a 245 ºC. A curva DSC do Polimorfo II apresentou dois eventos consecutivos, um endotérmico (Tpico = 193,9 ºC) e outro exotérmico (Tpico = 209,4 ºC), devido à fusão seguida de recristalização, a qual foi atribuída à conversão da forma II à forma I. Métodos termogravimétricos isotérmicos e não-isotérmicos foram empregados para determinar os parâmetros cinéticos do processo de decomposição térmica. Para experimentos não-isotérmicos, a energia de ativação (Ea) foi obtida a partir do gráfico de Log β vs 1/T, e os valores 154 e 123 kJ mol-1 foram encontrados, respectivamente, para os polimorfos I e II. Para os experimentos isotérmicos, a Ea foi obtida a partir do gráfico de lnt vs. 1/T a um nível de conversão constante. O valor médio encontrado foi 137 e 144 kJ mol-1, respectivamente, para a forma I e forma II.

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In this study, the kinetic behavior of Sf9 and Sf21 cells used in the production of a baculovirus biopesticide to control the pest of corn Spodoptera frugiperda was analyzed. Kinetic variables such as maximum specific growth rate, cell productivity, mean rate of infection, as well as the mean rate of occlusion body production were determined during the infection of these cell-lines with the extracellular virus of the S. frugiperda nucleopolyhedrovirus (SfMNPV). The Sf9 cell-line resulted in better viral production results (5.0 x 10(8) OB/mL) than the Sf21 cell-line (2.5 x 10(8) OB/mL).

Neste trabalho, analisou-se o comportamento cinético das células Sf9 e Sf21 utilizadas na produção de biopesticida para o controle de Spodoptera frugiperda. Variáveis cinéticas, como velocidade específica máxima de crescimento, produtividade em células, velocidade média de infecção e a velocidade média de produção de OB foram determinadas durante a infecção destas linhagens com o vírus extracelular do nucleopoliedrovirus de S. frugiperda. A linhagem Sf9 resultou em melhores resultados de produção do baculovírus (5 x 10(8) OB/mL), quando comparada à linhagem Sf21 (2,5 x 10(8) OB/mL) e outras linhagens da literatura.