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AIMS: The objective of this study was to determine the best conditions to produce invertase by Cunninghamella echinulata PA3S12MM and to immobilize and apply the enzyme. METHODS AND RESULTS: The maximum production was verified in 8 days of cultivation at 28°C supplemented with 10 g L-1 apple peel, reaching 1054.85 U ml-1 . The invertase was purified from the DEAE-Sephadex column. The derivative immobilized in alginate-gelatin-calcium phosphate showed reusability >50% for 19 cycles. The derivative immobilized in glutaraldehyde-chitosan showed greater thermostability and at a different pH. The hydrolysis of 15 ml of sucrose 500 g L-1 in a fixed bed reactor (total volume of 31 ml) produced 24.44 µmol min-1 of glucose and fructose at a residence time of 30 min and a conversion factor of 0.5. CONCLUSIONS: The new wild strain C. echinulata PA3S12MM presents high invertase production in medium supplemented with an agro-industrial residue and the immobilized enzyme showed high thermal stability and resistance at a different pH. SIGNIFICANCE AND IMPACT OF THE STUDY: The fungus C. echinulata PA3S12MM is an excellent producer of invertases in Vogel medium supplemented with apple peel. The enzyme is promising for industrial application since it has good performance in reusability and inverted sugar production.
Assuntos
Cunninghamella , beta-Frutofuranosidase , Cunninghamella/metabolismo , Estabilidade Enzimática , Enzimas Imobilizadas , Frutose , Concentração de Íons de Hidrogênio , Temperatura , beta-Frutofuranosidase/metabolismoRESUMO
Biotechnology offers innovative alternatives for industrial bioprocesses mainly because it uses enzymes that biodegrade the hemicellulose releasing fermentable sugars. Caulobacter crescentus (C. crescentus) has seven genes responsible for xylanolytic cleavage, 5 to ß-xylosidases (EC 3.2.1.37) and 2 for endoxylanases, like xynA2 (CCNA_03137) that encodes Xylanase II (EC 3.2.1.8) of the glycohydrolases-GH10 group. The xynA2 gene was amplified by PCR, cloned into the pTrcHisA vector e efficiently overexpressed in E. coli providing a His-tag fusion protein. Recombinant xylanase (XynA2) was purified by affinity chromatography using a nickel sepharose column and exhibited a single 43 kDa band on SDS-PAGE gel. XynA2 showed an optimum alkaline pH (8) and stability at alkaline pH for 24 h. Although C. crescentus is mesophilic, XynA2 has optimum temperature of 60 °C and is thermo-resistance at 65 °C. XynA maintains 66% of the enzymatic activity at high temperatures (90 °C) without being denatured.The enzyme displayed a xylanolitic activity free of cellulase to xylan from beechwood and it was not inhibited in the presence of 50 µmol mL-1 of xylose. In addition, dithiothreitol (DTT) induced XynA2 activity, as it improved its kinetic parameters by lowering the KM (5.78 µmol mL-1) and increasing the KCat/KM ratio (1.63 U s-1). Finally, C. crescentus XynA2 efficiently hydrolyzed corn straw with high release of reducing sugars that can be applied in different branches of the industry.
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Caulobacter crescentus/genética , Endo-1,4-beta-Xilanases/biossíntese , Endo-1,4-beta-Xilanases/isolamento & purificação , Biomassa , Cromatografia de Afinidade/métodos , Clonagem Molecular/métodos , Endo-1,4-beta-Xilanases/metabolismo , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Proteínas Recombinantes/genética , Especificidade por Substrato , Temperatura , Xilanos/metabolismo , XilosidasesRESUMO
The role of the clpB gene encoding HSP/chaperone ClpB was evaluated in the multiresistant antibiotic cells of Acinetobacter baumannii (RS4 strain) under stress-induced heat shock and different beta-lactams. The expression of the clpB gene was assessed by qPCR during heat shock at 45 °C and subinhibitory concentrations of ampicillin (30 µg mL-1), amoxicillin + sulbactam (8/12 µg mL-1), cefepime (30 µg mL-1), sulfamethoxazole + trimethoprim (120/8 µg mL-1) and meropenem (18 µg mL-1). The results indicated a transient increase in clpB transcription in all treatments except cefepime. Both in the presence of ampicillin and amoxicillin/sulbactam for 20 min, the mRNA-clpB synthesis was 1.4 times higher than that of the control at time zero. Surprisingly, the mRNA-clpB levels were more than 30-fold higher after 10 min of incubation with meropenem and more than eightfold higher in the presence of trimethoprim/sulfamethoxazole. In addition, western blot assays showed that the RS4 strain treated with meropenem showed a marked increase in ClpB protein expression. Our data indicate that during exposure to beta-lactams, A. baumannii adjusts the transcription levels of the clpB mRNA and protein to respond to stress, suggesting that the chaperone may act as a key cellular component in the presence of antibiotics in this bacterium.
Assuntos
Acinetobacter baumannii/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Genes Bacterianos , Resposta ao Choque Térmico/genética , Regulação para Cima/genética , beta-Lactamas/farmacologia , Acinetobacter baumannii/efeitos dos fármacos , Proteínas de Bactérias/metabolismo , Resposta ao Choque Térmico/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacosRESUMO
We present the unique case of a young female woman with an infected left liver hematoma as a complication of a previous endoscopic retrograde cholangiopancreatography for a choledocolithiasis. Urgent liver resections are rarely indicated and are usually performed in cases of important liver trauma. To our knowledge this is the first case of an urgent liver resection for the rare indication of late hepatic infected hematoma after an ERCP.
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Colangiopancreatografia Retrógrada Endoscópica , Hepatopatias , Feminino , Hemorragia Gastrointestinal , Hematoma/diagnóstico por imagem , Hematoma/etiologia , Hematoma/cirurgia , Humanos , Hepatopatias/diagnóstico por imagem , Hepatopatias/etiologia , Hepatopatias/cirurgiaRESUMO
Using the MoS2 -WTe2 heterostructure as a model system combined with electrochemical microreactors and density function theory calculations, it is shown that heterostructured contacts enhance the hydrogen evolution reaction (HER) activity of monolayer MoS2 . Two possible mechanisms are suggested to explain this enhancement: efficient charge injection through large-area heterojunctions between MoS2 and WTe2 and effective screening of mirror charges due to the semimetallic nature of WTe2 . The dielectric screening effect is proven minor, probed by measuring the HER activity of monolayer MoS2 on various support substrates with dielectric constants ranging from 4 to 300. Thus, the enhanced HER is attributed to the increased charge injection into MoS2 through large-area heterojunctions. Based on this understanding, a MoS2 /WTe2 hybrid catalyst is fabricated with an HER overpotential of -140 mV at 10 mA cm-2 , a Tafel slope of 40 mV dec-1 , and long stability. These results demonstrate the importance of interfacial design in transition metal dichalcogenide HER catalysts. The microreactor platform presents an unambiguous approach to probe interfacial effects in various electrocatalytic reactions.
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Paecilomyces variotii is a filamentous fungus that occurs worldwide in soil and decaying vegetation. Optimization of the fermentation process for exopolysaccharide (EPS) production from the fungus P. variotii, structure determination and immuno-stimulating activity of EPS were performed. Response surface methodology (RSM) coupled with central composite design (CCD) was used to optimize the physical and chemical factors required to produce EPS in submerged fermentation. Preliminary investigations to choose the three factors for the present work were made using a factorial experimental design. Glucose, ammonium nitrate (NH4NO3) and pH were used as variables for which, with constant temperature of 28 °C and agitation of 90 rpm, the optimal process parameters were determined as glucose values of 0.96%, NH4NO3 0.26% and pH 8.0. The three parameters presented significant effects. In this condition of culture, the main composition of the isolated EPS was a linear ß-(1 â 6)-linked-D-glucan, as determined by Nuclear Magnetic Resonance (NMR) and methylation analysis. This polysaccharide is a very unusual as an EPS from fungi, especially a filamentous fungus such as P. variotii. Murine peritoneal macrophages cultivated with ß-glucan for 6 and 48 h showed an increase in TNF-α, IL-6 and nitric oxide release with increased polysaccharide concentrations. Therefore, we conclude that the ß-(1 â 6)-linked-D-glucan produced in optimised conditions of P. variotii cultivation has an immune-stimulatory activity on murine macrophages.
Assuntos
Glucanos/metabolismo , Paecilomyces/metabolismo , Polissacarídeos Bacterianos/metabolismo , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância MagnéticaRESUMO
Insects are organisms extremely well adapted to diverse habitats, primarily due to their innate immune system, which provides them with a range of cellular and humoral responses against microorganisms. Lepidoptera hemolymph proteins involved in humoral responses are well known; however, there is a lack of knowledge about the sugarcane borer Diatraea saccharalis. In this present work, the hemolymph proteins of this pest insect were studied by applying proteomic methodologies. Two-dimensional electrophoresis (2-DE) gels of proteins extracted from naive larvae and larvae challenged with Escherichia coli (ATCC 11224) and Bacillus subtilis (ATCC 6623) showed an average of 300 spots, and 92 of these spots corresponded in all three 2-DE gels. Forty-one spots were excised and digested with trypsin and analyzed using mass spectrometry. After analysis, 10 proteins were identified, including some proteins of the immune system: ß-defensin-like protein, Turandot A-like protein, attacin-like protein, peptidoglycan recognition protein and cyclophilin-like protein. Nine proteins were present in both experimental conditions; however, ß-defensin-like protein was present only in hemolymph challenged by B. subtilis. Notably, attacin-like protein was strongly induced by challenge with E. coli, suggesting an immune response against the infection. However, antimicrobial activity was observed in the test zone of microbial growth inhibition of B. subtilis solely with the hemolymph extract of the larvae challenged with B. subtilis. We made for the first time a proteomic profile of the hemolymph of D. saccharalis in which it was possible to identify the presence of important proteins involved in the immune response.
Assuntos
Peptídeos Catiônicos Antimicrobianos/análise , Hemolinfa/microbiologia , Proteínas de Insetos/análise , Lepidópteros/microbiologia , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Bacillus subtilis/fisiologia , Proteínas de Transporte/análise , Proteínas de Transporte/metabolismo , Escherichia coli/fisiologia , Hemolinfa/fisiologia , Interações Hospedeiro-Patógeno , Proteínas de Insetos/metabolismo , Lepidópteros/fisiologia , ProteômicaRESUMO
The Caulobacter crescentus (NA1000) xynB5 gene (CCNA_03149) encodes a predicted ß-glucosidase-ß-xylosidase enzyme that was amplified by polymerase chain reaction; the product was cloned into the blunt ends of the pJet1.2 plasmid. Analysis of the protein sequence indicated the presence of conserved glycosyl hydrolase 3 (GH3), ß-glucosidase-related glycosidase (BglX) and fibronectin type III-like domains. After verifying its identity by DNA sequencing, the xynB5 gene was linked to an amino-terminal His-tag using the pTrcHisA vector. A recombinant protein (95 kDa) was successfully overexpressed from the xynB5 gene in E. coli Top 10 and purified using pre-packed nickel-Sepharose columns. The purified protein (BglX-V-Ara) demonstrated multifunctional activities in the presence of different substrates for ß-glucosidase (pNPG: p-nitrophenyl-ß-D-glucoside) ß-xylosidase (pNPX: p-nitrophenyl-ß-D-xyloside) and α-arabinosidase (pNPA: p-nitrophenyl-α-L-arabinosidase). BglX-V-Ara presented an optimal pH of 6 for all substrates and optimal temperature of 50 °C for ß-glucosidase and α-L-arabinosidase and 60 °C for ß-xylosidase. BglX-V-Ara predominantly presented ß-glucosidase activity, with the highest affinity for its substrate and catalytic efficiency (Km 0.24 ± 0.0005 mM, Vmax 0.041 ± 0.002 µmol min(-1) mg(-1) and Kcat/Km 0.27 mM(-1) s(-1)), followed by ß-xylosidase (Km 0.64 ± 0.032 mM, Vmax 0.055 ± 0.002 µmol min(-1) mg(-1) and Kcat/Km 0.14 mM(-1)s(-1)) and finally α-L-arabinosidase (Km 1.45 ± 0.05 mM, Vmax 0.091 ± 0.0004 µmol min(-1) mg(-1) and Kcat/Km 0.1 mM(-1) s(-1)). To date, this is the first report to demonstrate the characterization of a GH3-BglX family member in C. crescentus that may have applications in biotechnological processes (i.e., the simultaneous saccharification process) because the multifunctional enzyme could play an important role in bacterial hemicellulose degradation.
Assuntos
Caulobacter crescentus/enzimologia , Glicosídeo Hidrolases/metabolismo , Xilosidases/metabolismo , beta-Glucosidase/metabolismo , Caulobacter crescentus/genética , Clonagem Molecular , Biologia Computacional , Estabilidade Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Glicosídeo Hidrolases/genética , Concentração de Íons de Hidrogênio , Peso Molecular , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Especificidade por Substrato , Temperatura , Xilosidases/genética , beta-Glucosidase/genéticaRESUMO
The filamentous Thermoascus aurantiacus fungus characterized by its thermophilic nature, is recognized as an exceptional producer of various enzymes with biotechnological applications. This study aimed to explore biotechnological applications using polygalacturonase (PG) derived from the Thermoascus aurantiacus PI3S3 strain. PG production was achieved through submerged fermentation and subsequent purification via ion-exchange chromatography and gel filtration methods. The crude extract exhibited a diverse spectrum of enzymatic activities including amylase, cellulase, invertase, pectinase, and xylanase. Notably, it demonstrated the ability to hydrolyze sugarcane bagasse biomass, corn residue, and animal feed. The purified PG had a molecular mass of 36 kDa, with optimal activity observed at pH 4.5 and 70 °C. The activation energy (Ea) was calculated as 0.513 kJ mol-1, highlighting activation in the presence of Ca2+. Additionally, it displayed apparent Km, Vmax, and Kcat values of at 0.19 mg mL-1, 273.10 U mL-1, and 168.52 s-1, respectively, for hydrolyzing polygalacturonic acid. This multifunctional PG exhibited activities such as denim biopolishing, apple juice clarification, and demonstrated both endo- and exo-polygalacturonase activities. Furthermore, it displayed versatility by hydrolyzing polygalacturonic acid, carboxymethylcellulose, and xylan. The T. aurantiacus PI3S3 multifunctional polygalacturonase showed heightened activity under acidic pH, elevated temperatures, and in the presence of calcium. Its multifunctional nature distinguished it from other PGs, significantly expanding its potential for diverse biotechnological applications.
Assuntos
Saccharum , Thermoascus , Poligalacturonase/metabolismo , Thermoascus/metabolismo , Celulose , Enzimas Multifuncionais , Saccharum/metabolismo , Concentração de Íons de Hidrogênio , Estabilidade Enzimática , TemperaturaRESUMO
Endophytic fungi constitute a major part of the still unexplored fungal diversity and have gained interest as new biological sources of natural active compounds, including enzymes. Endophytic fungi were isolated from soybean leaves and initially screened on agar plates for the production of CMCase (carboxymethylcellulase), xylanase, amylase and protease. The highest Enzymatic Indexes (IE) were verified for xylanase (2.14 and 1.31) with the fungi M6-A6P5F2 and M12-A5P3F1.2 and CMCase (1.92 and 1.62) with the fungi M13-A9P2F1 and M12-A5P3F1.2, respectively. The production of xylanase and CMCase by the selected fungi was evaluated in submerged cultivation using beechwood xylan and carboxymethylcellulose (CMC), as well as sugarcane straw and bagasse in different ratios as carbon sources. Both types of lignocellulosic biomass proved to be good inducers of enzymatic activity. The best xylanase producer among the isolates was identified as Colletotrichum boninense. With this fungus, the highest xylanase activity was obtained with a sugarcane straw-bagasse mixture in a 50:50 ratio (383.63 U mL-1), a result superior to that obtained with the use of beechwood xylan (296.65 U mL-1). Regardingthe kinetic behavior of the crude xylanase, there was found optimal pH of 5.0 and optimal temperatures of 50°C and 60°C. At 40°C and 50°C, xylanase retained 87% and 76% of its initial catalytic activity, respectively. These results bring new perspectives on bioprospecting endophytic fungi for the production of enzymes, mainly xylanase, as well as the exploitation of agro-industrial by-products, such as sugarcane straw and bagasse.
Assuntos
Saccharum , Xilanos , Saccharum/microbiologia , Biomassa , FungosRESUMO
Developing accurate models for groundwater control is paramount for planning and managing life-sustaining resources (water) from aquifer reservoirs. Significant progress has been made toward designing and employing deep-forecasting models to tackle the challenge of multivariate time-series forecasting. However, most models were initially taught only to optimize natural language processing and computer vision tasks. We propose the Wavelet Gated Multiformer, which combines the strength of a vanilla Transformer with the Wavelet Crossformer that employs inner wavelet cross-correlation blocks. The self-attention mechanism (Transformer) computes the relationship between inner time-series points, while the cross-correlation finds trending periodicity patterns. The multi-headed encoder is channeled through a mixing gate (linear combination) of sub-encoders (Transformer and Wavelet Crossformer) that output trending signatures to the decoder. This process improved the model's predictive capabilities, reducing Mean Absolute Error by 31.26 % compared to the second-best performing transformer-like models evaluated. We have also used the Multifractal Detrended Cross-Correlation Heatmaps (MF-DCCHM) to extract cyclical trends from pairs of stations across multifractal regimes by denoising the pair of signals with Daubechies wavelets. Our dataset was obtained from a network of eight wells for groundwater monitoring in Brazilian aquifers, six rainfall stations, eleven river flow stations, and three weather stations with atmospheric pressure, temperature, and humidity sensors.
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Denim, also known as jeans, is a fabric made up of braided cotton threads dyed indigo blue, whose fibers contain approximately 10% of non-cellulosic impurities that reduce its commercial value. Microbial enzymes can act in the cleaning and desizing processes of jeans, improving their color, softness, and covering capacity. The recombinant Xylanase II (XynA2) from the aquatic bacterial Caulobacter crescentus (C. crescentus), previously characterized in terms of its biochemical features, was applied to the biotreatment of jeans to clean and degum it. The biotreatment performance was evaluated in terms of tissue weight loss, amount of reducing sugars released and analysis of the images obtained by scanning electron microscopy (SEM). Biotreated tissues, at 12 and 24 h, showed a dry weight loss of 4.9 and 6.6%, respectively. The reducing sugars amount released after XynA2 action over the jean's fibers showed statistically significant values when compared with each other and with their respective controls. SEM images clearly shown that the fabric treated for 12 h presented a smooth and polished surface, while the fabric treated for 24 h showed the cotton fibers broken, displaying severe damage to the textile. The best treatment for the jeans was in the presence of 1 U mg-1 XynA2 at pH 8 and 60 °C during 12 h. In conclusion, XynA2 of C. crescentus was satisfactorily applied for the biopolishing of denim jeans being a more sustainable alternative to the use of chemical and abrasive processes to obtain the same effects.
Assuntos
Caulobacter crescentus , Caulobacter crescentus/genética , Têxteis , Fibra de Algodão , Índigo Carmim , CorantesRESUMO
The xynB1 gene (CCNA 01040) of Caulobacter crescentus that encodes a bifunctional enzyme containing the conserved ß-Xylosidase and α-L-Arabinofuranosidase (ß-Xyl I-α-L-Ara) domains was amplified by PCR and cloned into the vector pJet1.2Blunt. The xynB1 gene was subcloned into the vector pPROEX-hta that produces a histidine-fused translation product. The overexpression of recombinant ß-Xyl I-α-L-Ara was induced with IPTG in BL21 (DE3) and the resulting intracellular protein was purified with pre-packaged nickel-Sepharose columns. The recombinant ß-Xyl I-α-L-Ara exhibited a specific ß-Xylosidase I activity of 1.25 U mg(-1) to oNPX and a specific α-L-Arabinofuranosidase activity of 0.47 U mg(-1) to pNPA. The predominant activity of the recombinant enzyme was its ß-Xylosidase I activity, and the enzymatic characterization was focused on it. The ß-Xylosidase I activity was high over the pH range 3-10, with maximal activity at pH 6. The enzyme activity was optimal at 45 °C, and a high degree of stability was verified over 240 min at this temperature. Moreover, ß-Xylosidase activity was inhibited in the presence of the metals Zn(2+) and Cu(2+), and the enzyme exhibited K(M) and V(Max) values of 2.89 ± 0.13 mM and 1.4 ± 0.04 µM min(-1) to oNPX, respectively. The modeled structure of ß-xylosidase I showed that its active site is highly conserved compared with other structures of the GH43 family. The increase in the number of contact residues responsible for maintaining the dimeric structure indicates that this dimer is more stable than the tetramer form.
Assuntos
Caulobacter crescentus/enzimologia , Caulobacter crescentus/genética , Xilosidases/metabolismo , Sequência de Aminoácidos , Clonagem Molecular , DNA Bacteriano/genética , Genes Bacterianos , Glicosídeo Hidrolases/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Estrutura Terciária de Proteína , Recombinação Genética , Análise de Sequência de DNA , Especificidade por Substrato , Xilosidases/genética , Xilosidases/isolamento & purificaçãoRESUMO
Complex systems in biology, climatology, medicine, and economy hold emergent properties such as non-linearity, adaptation, and self-organization. These emergent attributes can derive from large-scale relationships, connections, and interactive behavior despite not being apparent from their isolated components. It is possible to better comprehend complex systems by analyzing cross-correlations between time series. However, the accumulation of non-linear processes induces multiscale structures, therefore, a spectrum of power-law exponents (the fractal dimension) and distinct cyclical patterns. We propose the Multifractal detrended cross-correlation heatmaps (MF-DCCHM) based on the DCCA cross-correlation coefficients with sliding boxes, a systematic approach capable of mapping the relationships between fluctuations of signals on different scales and regimes. The MF-DCCHM uses the integrated series of magnitudes, sliding boxes with sizes of up to 5% of the entire series, and an average of DCCA coefficients on top of the heatmaps for the local analysis. The heatmaps have shown the same cyclical frequencies from the spectral analysis across different multifractal regimes. Our dataset is composed of sales and inventory from the Brazilian automotive sector and macroeconomic descriptors, namely the Gross Domestic Product (GDP) per capita, Nominal Exchange Rate (NER), and the Nominal Interest Rate (NIR) from the Central Bank of Brazil. Our results indicate cross-correlated patterns that can be directly compared with the power-law spectra for multiple regimes. We have also identified cyclical patterns of high intensities that coincide with the Brazilian presidential elections. The MF-DCCHM uncovers non-explicit cyclic patterns, quantifies the relations of two non-stationary signals (noise effect removed), and has outstanding potential for mapping cross-regime patterns in multiple domains.
Assuntos
Fractais , Meteorologia , Fatores de Tempo , Projetos de Pesquisa , BrasilRESUMO
The scientific community has been alarmed by the possible immunological evasion, higher infectivity, and severity of disease caused by the newest variants of SARS-CoV-2. The spike protein has an important role in the cellular invasion of viruses and is the target of several vaccines and therapeutic resources, such as monoclonal antibodies. In addition, some of the most relevant mutations in the different variants are on the spike (S) protein gene sequence that leads to structural alterations in the predicted protein, thus causing concern about the protection mediated by vaccines against these new strains. The present review highlights the most recent knowledge about COVID-19 and vaccines, emphasizing the different spike protein structures of SARS-CoV-2 and updating the reader about the emerging viral variants and their classifications, the more common viral mutations described and their distribution in Brazil. It also compiles a table with the most recent knowledge about all of the Omicron spike mutations.
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SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus/genética , Anticorpos Neutralizantes , Anticorpos Antivirais , COVID-19 , Humanos , Mutação , SARS-CoV-2/genética , Proteínas do Envelope Viral/genéticaRESUMO
Purified endoxylanase from Thermomyces lanuginosus PC7S1T was immobilized in calcium alginate, resulting in a yield of 78.5% and a reusability for 11 cycles. The stability of the immobilized enzyme was given for a pH range of 4 to 9 for 96 h. Endoxylanase immobilized in calcium alginate at 65 °C exhibited thermal stability equal to the soluble enzyme for 5 h, and at high temperatures of 75 °C and 85 °C showed half-lives of 4 and 3 h, respectively. Both soluble endoxylanase and immobilized forms were able to hydrolyze hemicellulose, obtained from low-lignin sorghum biomass pretreated with 5% H2O2 and 2% NaOH, after 1 h of incubation at 65 °C, releasing a mixture of short-chain xylooligosaccharides (X2-X6). The highest amounts of XOS generated were those for X5 (24 to 40%), X4 (33 to 39%), and X3 (11 to 22%). These XOS acted as prebiotics, promoting the growth of the probiotic L. acidophilus, similar to glucose in the MRS broth. These results show the potential of low-lignin sorghum to generate XOS with prebiotic activity, suggesting the application of these compounds in the food industry.
Assuntos
Endo-1,4-beta-Xilanases , Sorghum , Alginatos , Biomassa , Grão Comestível , Eurotiales , Glucuronatos , Peróxido de Hidrogênio , Hidrólise , Lignina/química , Oligossacarídeos/químicaRESUMO
The Cunninghamella echinulata PA3S12MM fungus is a great producer of invertases in a growth medium supplemented by apple peels. The enzyme was purified 4.5 times after two chromatographic processes, and it presented a relative molecular mass of 89.2 kDa. The invertase reached maximum activity at pH of 6 and at 60°C, in addition to presenting stability in alkaline pH and thermal activation at 50°C. The enzymatic activity increased in the presence of Mn2+ and dithiothreitol (DTT), while Cu2+ and Z2+ ions inhibited it. Also, DTT showed to protect enzymatic activity. The apparent values for Km , Vmáx , and Kcat for the sucrose hydrolysis were, respectively, 173.8 mmol/L, 908.7 mmol/L min-1 , and 1,388.79 s-1 . The carbohydrate content was of 83.13%. The invertase presented hydrolytic activity over different types of glycosidic bonds, such as α1 â 2ß (sucrose), α1 â 4 (polygalacturonic acid), α1 â 4 and α1 â 2 (pectin), and α1 â 1 (trehalose), indicating that the enzyme is multifunctional. Thus, the biochemical properties showed by the C. echinulata PA3S12MM suggest a broad industrial application, such as in the biomass hydrolysis or in the food industry. PRACTICAL APPLICATIONS: Invertases are hydrolytic enzymes employed in several industrial sectors. Given their great importance for the economy and several industrial sectors, there is a growing interest in microorganisms producing this enzyme. The analysis of the biochemical properties of invertase in C. echinulata PA3S12MM suggest applications in the food industry. Due to its increased hydrolytic activity, the hydrolysis process of the sucrose may employ invertase for the production of invert sugar. The stability at alkaline pH suggests an application in the development of enzymatic electrodes for the quantification of sucrose in food and beverage. The multifunctional activity may work in the biomass hydrolysis or saccharification of by-products for the extraction of fermentable sugars. The high level of invertase N-linked glycosylation of invertase grants this enzyme thermal stability at high temperatures, in addition to resistance against the action of proteases, which are desirable characteristics for the application of this enzyme in industrial processes.
Assuntos
Cunninghamella , beta-Frutofuranosidase , Concentração de Íons de Hidrogênio , TemperaturaRESUMO
Despite the outstanding progresses in Multiple Myeloma treatment options in the last decades, it remains an incurable disease nowadays. Infectious events are a complication due to an impaired immune system associated with MM, sometimes a life-threatening one, particularly on the first months after the diagnosis. Both the underlying disease and treatment can contribute to the infection risk, so a biomarker that assess this risk could be highly relevant for a more tailored management of the patient. The measurement of the heavy+light chain (HLC) pairs of immunoglobulins in serum allows the quantification of both the monoclonal component and the non-monoclonal immunoglobulin of the same isotype. This approach has demonstrated high sensitivity for the detection of the clonality and prognostic value for MM. HLC pair suppression itself has prognostic power and it has been proposed to be a reflection of the immune system' attempt to control the tumor. In this study we evaluated the impact of the HLC pair suppression on the rate of bloodstream infections (BSI) and early death in 115 newly diagnosed MM patients. Twenty-one percent of the patients suffered a BSI in the first 6 months after diagnosis, of which 58% died within this period, accounting to 67% of the early deaths in global and highlighting the major impact of infections on MM patients in a "real world" setting. Severe HLC pair suppression identified patients with a higher risk of early BSI (HR: 6,97, p=0,009), and extreme HLC pair suppression together with BSI event and age >65 were independent risk factors for early death (p<0,001). Based on these factors, a stratification model was generated to allow identify patients at a higher risk of early death and poorer OS, with an apparently better performance than the ISS on the early death context. In conclusion, HLC pair suppression associates with both a higher risk of life-threatening early infection and early death in newly diagnosed MM patients. Patients older than 65 with extreme HLC pair suppression and BSI are at a high risk of early death, and thus patients presenting with these criteria have a very adverse prognosis.
RESUMO
INTRODUCTION: The etiologies of most chronic temporomandibular joint disorders are unknown. However, an association between habitual chewing on a particular side and chronic temporomandibular joint disorders has been reported. The aim of this study was to investigate the differences between sides (affected vs unaffected) of biodynamic factors (including lateral dental guidance determined by dental anatomy) or condylar path angles (determined by temporomandibular joint morphology) and chewing function (physiological alternate chewing vs single habitual chewing side). The study scope was to investigate possible etiological factors to improve the understanding of temporomandibular joint disorders. The null hypothesis was that no difference would be found between sides that are or are not affected by chronic temporomandibular joint disorders in chewing function or in levels of dental or temporomandibular joint remodeling. METHODS: This cross-sectional, double-blind study involved 24 adults with substantial, chronic, unilateral symptoms diagnosed as temporomandibular joint disorders. Chewing function, temporomandibular joint remodeling (using axiography) and dental anatomy (lateral guidance angles using kinesiography) were assessed. RESULTS: Habitual chewing on one particular side was observed in 17 of 24 participants; significantly more (n=15) chewed on the affected side than on the unaffected side (P=0.002 in a two-tailed Fisher's exact test; risk estimate=4.5; 95% CI 1.326-15.277). The condylar path (CP) angle was steeper on the affected side than on the unaffected side (mean (standard deviation)=50.52° (9.98°) versus 45.50° (7.98°); P=0.002 in a two-tailed t-test). The lateral guidance (LG) angles were flatter on the affected side in all 24 participants. CONCLUSION: Our results suggest that habitual chewing on one side may be associated with increasing condylar path, with flattening lateral guidance angles, and also with chronic temporomandibular joint disorder on the habitual chewing side.
Assuntos
Mastigação , Transtornos da Articulação Temporomandibular , Adulto , Estudos Transversais , Método Duplo-Cego , Humanos , Côndilo Mandibular , Articulação TemporomandibularRESUMO
Despite tremendous progress being made in recent years, multiple myeloma (MM) remains a challenging disease. The laboratory plays a critical role in the overall management of patients. The diagnosis, prognosis, clinical monitoring and evaluation of the response are key moments in the clinical care process. Conventional laboratory methods have been and continue to be the basis of laboratory testing in monoclonal gammopathies, along with the serum free light chain test. However, more accurate methods are needed to achieve new and more stringent clinical goals. The heavy/light chain assay is a relatively new test which can overcome some of the limitations of the conventional methods for the evaluation of intact immunoglobulin MM patients. Here, we report an update of the evidence accumulated in recent years on this method regarding its use in MM.