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Cardiovascular diseases, resulting from the deposition of clots in blood vessels, are the leading cause of death worldwide. Fibrinolytic enzymatic activity can catalyze blood clot degradation. Findings show that 36 fungal isolates recovered from Caatinga soils have the potential to produce fibrinolytic protease under submerged conditions. About 58 % of the isolates displayed fibrinolytic activity above 100 U/mL, with Mucor subtilissimus UCP 1262 being the most active. The protease was biochemically and biophysically characterized, showing that the enzyme had a high affinity for SAApNA substrate and was significantly inhibited by fluoride methyl phenyl sulfonyl-C7H7FO2S, suggesting that it is a chymotrypsin-like serine protease. The highest enzyme activity was detected at pH 5.0 and 28 °C. This fibrinolytic protease's far-UV circular dichroism (CD) showed that its secondary structure was primarily α-helical. The purified fibrinolytic enzyme may represent a novel therapeutic agent for treating thrombosis. At temperatures above 65 °C, the enzyme lost all its secondary structure. Its melting temperature was 58.1 °C, the denaturation enthalpy 85.1 kcal/mol, and the denaturation entropy 0.26 kcal/Kâmol.
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Mucor , Mucor/enzimología , Concentración de Iones de Hidrógeno , Dicroismo Circular , Microbiología del Suelo , Péptido Hidrolasas/química , Péptido Hidrolasas/aislamiento & purificación , Péptido Hidrolasas/metabolismo , Temperatura , Fibrinolíticos/química , FibrinólisisRESUMEN
Eleocharis R. Br. (Cyperaceae) species are known for having holocentric chromosomes, which enable rapid karyotype differentiation. High intra- and interspecific variations in chromosome numbers and genome sizes are documented for different Eleocharis species, frequently accompanied by fluctuations in the repetitive DNA fraction. However, a lack of detailed analysis has hampered a better understanding of the interplay between holocentricity and repetitive DNA evolution in this genus. In our study, we confirmed the holocentricity of Eleocharis chromosomes by immunostaining against the kinetochore protein KNL1 and the cell-cycle dependent posttranslational modifications histone H2AThr121ph and H3S10ph. We further studied the composition and chromosomal distribution of the main satellite DNA repeats found in the newly sequenced species E. maculosa, E. geniculata, E. parodii, E. elegans, and E. montana. Five of the six satellites discovered were arranged in clusters, while EmaSAT14 was distributed irregularly along the chromatid length in a line-like manner. EmaSAT14 monomers were present in a few copies in few species across the Eleocharis phylogenetic tree. Nonetheless, they were accumulated within a restricted group of Maculosae series, subgenus Eleocharis. The data indicates that the amplification and line-like distribution of EmaSAT14 along chromatids may have occurred recently within a section of the genus.
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Perivascular adipose tissue (PVAT) negatively regulates vascular muscle contraction. However, in the context of obesity, the PVAT releases vasoconstrictor substances that detrimentally affect vascular function. A pivotal player in this scenario is the peptide endothelin-1 (ET-1), which induces oxidative stress and disrupts vascular function. The present study postulates that obesity augments ET-1 production in the PVAT, decreases the function of the nuclear factor erythroid 2-related factor-2 (Nrf2) transcription factor, further increasing reactive oxygen species (ROS) generation, culminating in PVAT dysfunction. Male C57BL/6 mice were fed either a standard or a high-fat diet for 16 weeks. Mice were also treated with saline or a daily dose of 100 mg·kg-1 of the ETA and ETB receptor antagonist Bosentan, for 7 days. Vascular function was evaluated in thoracic aortic rings, with and without PVAT. Mechanistic studies utilized PVAT from all groups and cultured WT-1 mouse brown adipocytes. PVAT from obese mice exhibited increased ET-1 production, increased ECE1 and ETA gene expression, loss of the anticontractile effect, as well as increased ROS production, decreased Nrf2 activity, and downregulated expression of Nrf2-targeted antioxidant genes. PVAT of obese mice also exhibited increased expression of Tyr216-phosphorylated-GSK3ß and KEAP1, but not BACH1 - negative Nrf2 regulators. Bosentan treatment reversed all these effects. Similarly, ET-1 increased ROS generation and decreased Nrf2 activity in brown adipocytes, events mitigated by BQ123 (ETA receptor antagonist). These findings place ET-1 as a major contributor to PVAT dysfunction in obesity and highlight that pharmacological control of ET-1 effects restores PVAT's cardiovascular protective role.
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Tejido Adiposo , Regulación hacia Abajo , Endotelina-1 , Ratones Endogámicos C57BL , Factor 2 Relacionado con NF-E2 , Obesidad , Especies Reactivas de Oxígeno , Animales , Endotelina-1/metabolismo , Obesidad/metabolismo , Obesidad/fisiopatología , Masculino , Tejido Adiposo/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Bosentán/farmacología , Dieta Alta en Grasa , Ratones , Estrés Oxidativo , Receptor de Endotelina A/metabolismo , Receptor de Endotelina A/genética , Enzimas Convertidoras de Endotelina/metabolismo , Aorta Torácica/metabolismo , Aorta Torácica/efectos de los fármacos , Aorta Torácica/fisiopatologíaRESUMEN
Cardiovascular diseases (CVDs) cause 30% of deaths each year, and in 2030, around 23.6 million people will die due to CVDs. The major challenge is to obtain molecules with minimal adverse reactions that can prevent and dissolve blood clots. In this context, fibrinolytic enzymes from diverse microorganism sources have been extensively investigated due to their potential to act directly and specifically on the fibrin clot, preventing side effects and performing potential thrombolytic effects. However, most researches focus on the purification and characterization of proteases, with little emphasis on the mechanism of action and pharmacological characteristics, including toxicity assays which are essential to assess safety and side effects. Therefore, this work aims to emphasize the importance of evaluations indicating the toxicological profile of fibrinolytic proteases through in vitro and in vivo tests. Both types of assays contribute as preclinical stage in drug development and are crucial for clinical applications. This scarcity creates arbitrary barriers to further studies. This work should further encourage the development of studies to ensure the safety and effectivity of fibrinolytic proteases.
Suggested pre-clinical trials aim to validate more specific methods for fibrinolytic enzymes;Current toxicity standards can be adapted to better assess the profile of fibrinolytic enzymes;The class of fibrinolytic enzymes must be carefully evaluated according to the method of application.
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High levels of testosterone (Testo) are associated with cardiovascular risk by increasing reactive oxygen species (ROS) formation. NADPH oxidases (NOX) are the major source of ROS in the vasculature of cardiovascular diseases. NOX4 is a unique isotype, which produces hydrogen peroxide (H2O2), and its participation in cardiovascular biology is controversial. So far, it is unclear whether NOX4 protects from Testo-induced endothelial injury. Thus, we hypothesized that supraphysiological levels of Testo induce endothelial NOX4 expression to attenuate endothelial injury. Human mesenteric vascular endothelial cells (HMECs) and human umbilical vein endothelial cells (HUVEC) were treated with Testo (10-7 M) with or without a NOX4 inhibitor [GLX351322 (10-4 M)] or NOX4 siRNA. In vivo, 10-wk-old C57Bl/6J male mice were treated with Testo (10 mg/kg) for 30 days to study endothelial function. Testo increased mRNA and protein levels of NOX4 in HMECs and HUVECs. Testo increased superoxide anion (O2-) and H2O2 production, which were abolished by NOX1 and NOX4 inhibition, respectively. Testo also attenuated bradykinin-induced NO production, which was further impaired by NOX4 inhibition. In vivo, Testo decreased H2O2 production in aortic segments and triggered endothelial dysfunction [decreased relaxation to acetylcholine (ACh)], which was further impaired by GLX351322 and by a superoxide dismutase and catalase mimetic (EUK134). Finally, Testo led to a dysregulated endothelial cell migration, which was exacerbated by GLX351322. These data indicate that supraphysiological levels of Testo increase the endothelial expression and activity of NOX4 to counterbalance the deleterious effects caused by Testo in endothelial function.NEW & NOTEWORTHY By inducing ROS formation, high levels of testosterone play a major role in the pathogenesis of cardiovascular disease. NOXs are the major sources of ROS in the vasculature of cardiovascular diseases. Herein, we describe a novel compensatory mechanism by showing that NOX4 is a protective oxidant enzyme and counterbalances the deleterious effects of testosterone in endothelial cells by modulating hydrogen peroxide formation.
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Movimiento Celular , Endotelio Vascular , Células Endoteliales de la Vena Umbilical Humana , Peróxido de Hidrógeno , Ratones Endogámicos C57BL , NADPH Oxidasa 4 , Testosterona , Animales , Humanos , Masculino , Ratones , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Células Endoteliales/metabolismo , Células Endoteliales/efectos de los fármacos , Endotelio Vascular/metabolismo , Endotelio Vascular/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/farmacología , NADPH Oxidasa 4/metabolismo , NADPH Oxidasa 4/genética , Especies Reactivas de Oxígeno/metabolismo , Testosterona/farmacología , Testosterona/metabolismoRESUMEN
The decline in the quality of water resources in the Amazon is very rapid in cities suffering from unplanned urban growth. The region has two defined seasons, winter (wet) and summer (dry), which directly affect the behavior of contaminants in aquatic ecosystems. The aim of this study was to assess the ecological and human health risks associated with the use of the watershed. In addition, an ecological index was proposed: the Quality Index for Aquatic Life, for the risk of contaminants to aquatic life. Sampling was carried out at six points in the Juá watershed. Physicochemical parameters, major anions, metals and total phosphorus were analyzed at both stations between 2020 and 2021. The highest concentrations of contaminants were found in the rainy season, due to the washing away of the banks. In this sense, Cl presented a concentration more than 307 times higher than that permitted by Brazilian legislation (wet). The ecological index showed that the watershed has a high risk of metals such as Cr III and Cr VI for the biota. The human health risk analysis showed a low risk; however, the lack of basic sanitation in the city indicates that monitoring of urban water resources is necessary.
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Metales Pesados , Contaminantes Químicos del Agua , Humanos , Calidad del Agua , Monitoreo del Ambiente , Ecosistema , Contaminantes Químicos del Agua/análisis , Medición de Riesgo , Metales Pesados/análisis , Ríos , ChinaRESUMEN
Snake venoms contain various bradykinin-potentiating peptides (BPPs). First studied for their vasorelaxant properties due to angiotensin converting enzyme (ACE) inhibition, these molecules present a range of binding partners, among them the argininosuccinate synthase (AsS) enzyme. This has renewed interest in their characterization from biological sources and the evaluation of their pharmacological activities. In the present work, the low molecular weight fraction of Bothrops moojeni venom was obtained and BPPs were characterized by mass spectrometry. Eleven BPPs or related peptides were sequenced, and one of them, BPP-Bm01, was new. Interestingly, some oxidized BPPs were detected. The three most abundant peptides were BPP-Bm01, BPP-Bax12, and BPP-13a, and their putative interactions with the AsS enzyme were investigated in silico. A binding cavity for these molecules was predicted, and docking studies allowed their ranking. Three peptides were synthesized and submitted to vasorelaxation assays using rat aortic rings. While all BPPs were active, BPP-Bm01 showed the highest potency in this assay. This work adds further diversity to BPPs from snake venoms and suggests, for the first time, a putative binding pocket for these molecules in the AsS enzyme. This can guide the design of new and more potent AsS activators.
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Aorta , Bothrops , Oligopéptidos , Péptidos , Serpientes Venenosas , Animales , Ratas , Brasil , Aorta/efectos de los fármacos , Péptidos/farmacología , Péptidos/química , Bradiquinina/farmacología , Masculino , Venenos de Crotálidos/farmacología , Venenos de Crotálidos/química , Ratas Wistar , Venenos de Serpiente/farmacología , Vasodilatadores/farmacología , Vasodilatadores/química , Estructura MolecularRESUMEN
Mushrooms are a source of primary and secondary metabolites. Little is known about the most suitable conditions for production of mushrooms by submerged fermentation. This article reports antioxidant and cytotoxic assays, in addition to quantitatively evaluating the content of proteases with fibrinolytic action in the crude extracts of two species of edible mushrooms produced in different formulations, as well as evaluating the recovery of these enzymes by aqueous two-phase systems (ATPS). The mushrooms Pleurotus ostreatus and Pleurotus eryngii, at concentration of 100 µg/mL, displayed inhibition of DPPH and ABTS radicals below 50%. In the cytotoxicity test, the cells human fibroblast cell lines (MRC-5) showed cell viability greater than 80%. Concerning fibrinolytic activity, P. eryngii presented 226.47 ± 7.26 U/mL, therefore being more efficient than P. ostreatus (71.5 ± 0.56 U/mL). In the recovery of the P. eryngii extract by ATPS, the fibrinolytic protease was partitioned in the salt phase (30.25 U/mL). The molecular mass of the proteases was between 75 and 100 kDa. These results prove the low cytotoxicity of the extracts produced and that fermentation in supplemented malt broth favored the excretion of fibrinolytic proteases compared to the other evaluated media.
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Agaricales , Antineoplásicos , Pleurotus , Humanos , Antioxidantes/química , Pleurotus/química , Péptido Hidrolasas/metabolismo , Agaricales/química , Endopeptidasas/metabolismo , Antineoplásicos/metabolismoRESUMEN
Collagenases are proteases able to degrade native and denatured collagen, with broad applications such as leather, food, and pharmaceutical industries. The aim of this research was to purify and characterize a collagenase from Streptomyces antibioticus. In the present work, the coffee ground substrate provided conditions to obtaining high collagenase activity (377.5 U/mL) using anion-exchange DEAE-Sephadex G50 chromatographic protocol. SDS-PAGE revealed the metallo-collagenase with a single band of 41.28 kDa and was able to hydrolyzed type I and type V collagen producing bioactive peptides that delayed the coagulation time. The enzyme activity showed stability across a range of pH (6.0-11) and temperature (30-55 °C) with optima at pHâ¯7.0 and 60 °C, respectively. Activators include Mg+2, Ca+2, Na+, K+, while full inhibition was given by other tested metalloproteinase inhibitors. Kinetic parameters (Km of 27.14 mg/mol, Vmax of 714.29 mg/mol/min, Kcat of 79.9 s-1 and Kcat/Km of 2.95 mL/mg/s) and thermodynamic parameters (Ea of 65.224 kJ/mol, ΔH of 62.75 kJ/mol, ΔS of 1.96 J/mol, ΔG of 62.16 kJ/mol, ΔGE-S of 8.18 kJ/mol and ΔGE-T of -2.64 kJ/mol) were also defined. Coffee grounds showed to be an interesting source to obtaining a collagenase able to produce bioactive peptides with anticoagulant activity.
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Streptomyces antibioticus , Café , Termodinámica , Colagenasas , Péptidos , Concentración de Iones de Hidrógeno , CinéticaRESUMEN
AIMS: Overproduction of reactive oxygen species (ROS) is a pathologic hallmark of cyclophosphamide toxicity. For this reason, antioxidant compounds emerge as promising tools for preventing tissue damage induced by cyclophosphamide. We hypothesized that melatonin would display cytoprotective action in the vasculature by preventing cyclophosphamide-induced oxidative stress. MATERIALS AND METHODS: Male C57BL/6 mice (22-25 g) were injected with a single dose of cyclophosphamide (300 mg/kg; i.p.). Mice were pretreated or not with melatonin (10 mg/kg/day, i.p.), given during 4 days before cyclophosphamide injection. Functional (vascular reactivity) and oxidative/inflammatory patterns were evaluated at 24 h in resistance arteries. The antioxidant action of melatonin was assessed in vitro in cultured vascular smooth muscle cells (VSMCs) of mesenteric arteries. KEY FINDINGS: Cyclophosphamide induced ROS generation in both mesenteric arterial bed (MAB) and cultured VSMCs, and this was normalized by melatonin. Cyclophosphamide-induced ROS generation and lipoperoxidation in the bladder and kidney was also prevented by melatonin. Increased levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 were detected in the MAB of cyclophosphamide-treated mice, all of which were prevented by melatonin. Functional assays using second-order mesenteric arteries of cyclophosphamide-treated mice revealed a decrease in vascular contractility. Melatonin prevented vascular hypocontractility in the cyclophosphamide group. Melatonin partially prevented the decrease in myeloperoxidase (MPO) and N-acetyl-beta-D-glucosaminidase (NAG) activities in the MAB of the cyclophosphamide group. SIGNIFICANCE: Melatonin may constitute a novel and promising therapeutic approach for management of the toxic effects induced by cyclophosphamide in the vasculature.
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Melatonina , Ratones , Masculino , Animales , Especies Reactivas de Oxígeno/farmacología , Melatonina/uso terapéutico , Antioxidantes/metabolismo , Ratones Endogámicos C57BL , Ciclofosfamida/toxicidad , Estrés Oxidativo , Arterias Mesentéricas/metabolismoRESUMEN
BACKGROUND: Renin-angiotensin (Ang II)-aldosterone system (RAAS) is crucial for the cardiovascular risk associated with excessive ethanol consumption. Disturbs in mitochondria have been implicated in multiple cardiovascular diseases. However, if mitochondria dysfunction contributes to ethanol-induced vascular dysfunction is still unknown. We investigated whether ethanol leads to vascular dysfunction via RAAS activation, mitochondria dysfunction, and mitochondrial reactive oxygen species (mtROS). METHODS: Male C57/BL6J or mt-keima mice (6-8-weeks old) were treated with ethanol (20% vol./vol.) for 12 weeks with or without Losartan (10 mg/kg/day). RESULTS: Ethanol induced aortic hypercontractility in an endothelium-dependent manner. PGC1α (a marker of biogenesis), Mfn2, (an essential protein for mitochondria fusion), as well as Pink-1 and Parkin (markers of mitophagy), were reduced in aortas from ethanol-treated mice. Disturb in mitophagy flux was further confirmed in arteries from mt-keima mice. Additionally, ethanol increased mtROS and reduced SOD2 expression. Strikingly, losartan prevented vascular hypercontractility, mitochondrial dysfunction, mtROS, and restored SOD2 expression. Both MnTMPyP (SOD2 mimetic) and CCCP (a mitochondrial uncoupler) reverted ethanol-induced vascular dysfunction. Moreover, L-NAME (NOS inhibitor) and EUK 134 (superoxide dismutase/catalase mimetic) did not affect vascular response in ethanol group, suggesting that ethanol reduces aortic nitric oxide (NO) and H2O2 bioavailability. These responses were prevented by losartan. CONCLUSION: AT1 receptor modulates ethanol-induced vascular hypercontractility by promoting mitochondrial dysfunction, mtROS, and reduction of NO and H2O2 bioavailability. Our findings shed a new light in our understanding of ethanol-induced vascular toxicity and open perspectives of new therapeutic approaches for patients with disorder associated with abusive ethanol drinking.
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Losartán , Lesiones del Sistema Vascular , Humanos , Ratones , Masculino , Animales , Losartán/farmacología , Receptor de Angiotensina Tipo 1/metabolismo , Etanol/toxicidad , Peróxido de Hidrógeno/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Mitocondrias/metabolismoRESUMEN
Structural karyotype changes result from ectopic recombination events frequently associated with repetitive DNA. Although most Phaseolus species present relatively stable karyotypes with 2n = 22 chromosomes, the karyotypes of species of the Leptostachyus group show high rates of structural rearrangements, including a nested chromosome fusion that led to the dysploid chromosome number of the group (2n = 20). We examined the roles of repetitive landscapes in the rearrangements of species of the Leptostachyus group using genome-skimming data to characterize the repeatome in a range of Phaseolus species and compared them to species of that group (P. leptostachyus and P. macvaughii). LTR retrotransposons, especially the Ty3/gypsy lineage Chromovirus, were the most abundant elements in the genomes. Differences in the abundance of Tekay, Retand, and SIRE elements between P. macvaughii and P. leptostachyus were reflected in their total amounts of Ty3/gypsy and Ty1/copia. The satellite DNA fraction was the most divergent among the species, varying both in abundance and distribution, even between P. leptostachyus and P. macvaughii. The rapid turnover of repeats in the Leptostachyus group may be associated with the several rearrangements observed.
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Phaseolus , Phaseolus/genética , ADN de Plantas/genética , ADN Satélite/genética , Retroelementos , Filogenia , Genoma de Planta , Evolución MolecularRESUMEN
KEY MESSAGE: Karyotypes evolve through numerical and structural chromosome rearrangements. We show that Phaseolus leptostachyus, a wild bean, underwent a rapid genome reshuffling associated with the reduction from 11 to 10 chromosome pairs, but without whole genome duplication, the highest chromosome evolution rate known for plants. Plant karyotypes evolve through structural rearrangements often associated with polyploidy or dysploidy. The genus Phaseolus comprises ~ 90 species, five of them domesticated due to their nutritional relevance. Most of the species have 2n = 22 karyotypes and are highly syntenic, except for three dysploid karyotypes of species from the Leptostachyus group (2n = 20) that have accumulated several rearrangements. Here, we investigated the degrees of structural rearrangements among Leptostachyus and other Phaseolus groups by estimating their chromosomal evolution rates (CER). For this, we combined our oligo-FISH barcode system for beans and chromosome-specific painting probes for chromosomes 2 and 3, with rDNA and a centromeric probe to establish chromosome orthologies and identify structural rearrangements across nine Phaseolus species. We also integrated the detected rearrangements with a phylogenomic approach to estimate the CERs for each Phaseolus lineage. Our data allowed us to identify translocations, inversions, duplications and deletions, mostly in species belonging to the Leptostachyus group. Phaseolus leptostachyus showed the highest CER (12.31 rearrangements/My), a tenfold increase in contrast to the 2n = 22 species analysed. This is the highest rate known yet for plants, making it a model species for investigating the mechanisms behind rapid genome reshuffling in early species diversification.
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Phaseolus , Phaseolus/genética , Cariotipo , Cariotipificación , Centrómero , Reordenamiento GénicoRESUMEN
Feedlot finishing of goats is a growing practice, but the economic viability of this technology is compromised by the inclusion of ingredients such as corn and soybean. An alternative to minimize this barrier is the use of agroindustry coproducts as substitutes for those ingredients, such as crude glycerol. This study aimed to evaluated the metabolism of crossbred Boer finishing goats fed diets containing crude glycerin from biodiesel production. Thirty-two crossbred, castrated goat of age were distributed in a fully randomized experimental design with four treatments and eight replicates. The experiment lasted 69 days, and goats were fed sorghum silage and concentrate, with the inclusion of crude glycerin in the diet at levels of 0, 50, 100, and 150 g/kg on a dry matter basis. The diets did not have an effect (p > 0.05) on the serum urea levels. Increasing dietary crude glycerin levels did not the influence the metabolic or urinary profiles (p > 0.05). The liver tissue of the goats fed diets containing the highest crude glycerin inclusion levels showed deleterious effects. The inclusion of crude glycerin with approximately 6.6 g/kg methanol caused deleterious effects to the liver tissue of Boer crossbred goats as the glycerin concentrations increased. However, glycerin levels did not cause deleterious effects on the liver tissue or on the serum or urinary profiles. The use of crude glycerin with lower methanol content is recommended for goat diets.
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A fibrinolytic enzyme from the microalga Dunaliella tertiolecta was produced under mixotrophic conditions using different corn steep liquor (CSL) concentrations ( 0 ≤ CLS ≤ 0.75%), purified using a combination of salting out and ion-exchange chromatography, and then biochemical characterized. Cultivation of this microalga using 0.5% CSL led to the highest maximum cell concentration (1.960±0.010 mg L-1) and cell productivity (0.140g L-1 day-1), besides a high fibrinolytic activity of the extract obtained by the homogenization method (102 ±1 U mL-1). The enzyme extracted from the microalgal biomass was 5-fold purified with a 20% yield and was found to have a specific activity of 670 U mg-1. The enzyme, whose molecular weight determined by fibrin zymography was 10 kDa, was shown to be stable at pH 3.0-9.0 and up to 70°C with optimal pH and temperature values of 8.0 and 50°C, respectively. When compared to other fibrinolytic enzymes, this protease stood out for its high fibrinolytic activity, which was enhanced by Fe2+, inhibited by Zn2+, Cu2+, Mg2+, and Ca2+, and strongly inhibited by phenylmethylsulfonyl fluoride, suggesting that it belongs to the serine metalloprotease family. Moreover, thanks to its thermal stability, the enzyme may be easily preserved and activated under high-temperature conditions.
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Microalgas , Zea mays , Análisis Costo-Beneficio , Fibrina , Concentración de Iones de Hidrógeno , TemperaturaRESUMEN
The emergence of SARS-CoV-2 variants can affect their detection via RT-qPCR. The Omicron variant has a greater affinity for the upper respiratory system and causes clinical conditions similar to bronchitis, as opposed to the pneumonitis-like conditions caused by other SARS-CoV-2 variants. This characteristic increases the viscosity of clinical samples collected for diagnosis. Coinciding with the arrival of the Omicron variant, we observed a failure in control gene expression in our laboratory. In this report, we have optimized a rapid nucleic acid extraction step to restore gene expression and detect the presence of the SARS-CoV-2 virus. We reevaluated 3000 samples, compared variant types occurring in different time periods, and confirmed that the presence of the Omicron variant was responsible for changes observed in the characteristics of these clinical samples. For samples with large amounts of mucus, such as those containing the Omicron variant, a modification to the nucleic acid extraction step was sufficient to restore the quality of RT-qPCR results.
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COVID-19 , Ácidos Nucleicos , Humanos , Brasil , SARS-CoV-2/genética , COVID-19/diagnóstico , ARNRESUMEN
This study was carried out to evaluate the effect of ensiling sorghum silage with urea and amending the aerobic exposure nutrients intake and apparent digestibility, ingestive behaviour and blood serum metabolites of feedlot lambs. Forty uncastrated crossbred Dorper × Santa Inês lambs, aged 150 ± 15 days and with an initial body weight of 21.73 ± 2.40 kg, were used. Animals were assigned in a 2 × 3 factorial arrangement. Thus, six silage diets were produced with various urea addition levels (UA: 0 and 5 g/kg on a natural matter basis) and periods of aerobic exposure of silages (PAE: 0, 24 and 48 h). An effect was observed for nutrient intakes of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fibre corrected for ash and protein (NDFap) and total digestive nutrients TDN (g/day) and for the total apparent digestibility of DM, OM and CP. There was an interaction effect between urea levels and aerobic exposure for ether extract (EE) and NDFap intakes (g/kg) and nonfibrous carbohydrate (NFC) digestibility (g/kg) (p = 0.012). The addition of 5 g/kg of urea to sorghum ensilage improved the digestibility parameters without changing dry matter intake and ingestive behaviour. The addition or not of urea does not change the blood parameters of the animals.
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Background: Traumatic brain injury (TBI) has a high economic and social impact on the family dynamics, particularly among children. High-quality and comprehensive epidemiological studies about TBI in this population are limited worldwide, specifically in Latin America. Therefore, this study aimed to elucidate the epidemiology of TBI among children in Brazil and its effects on the public health system. Methods: This epidemiological (cohort) retrospective study collected data from the Brazilian healthcare database between 1992 and 2021. Results: The mean annual volume of hospital admission (HA) due to TBI in Brazil was 29,017. Moreover, the incidence of TBI in the paediatric population was 45.35 admissions per 100,000 inhabitants/year. Furthermore, approximately 941 paediatric hospital deaths per year were caused by TBI, with an in-hospital lethality rate of 3.21%. The average annual financial transfer for TBI was 12,376,628 USD, and the mean cost per admission was 417 USD. In addition, the mean length of hospital stay was 4.2 days. Notably, the length of stay in the hospital was longer among males, Afro-Brazilians patients and individuals aged 15-19 years. Conclusion: Paediatric TBI is an important public health issue worldwide with high social and economic costs. The incidence of paediatric TBI in Brazil is similar to that in developing countries. Moreover, male predominance (2.3:1) was observed in relation paediatric TBI. Notably, during the pandemic, the incidence of paediatric HA has decreased. To the best of our knowledge, this is the first epidemiological study that specifically evaluates paediatric TBI in Latin America.
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The aim of this study was to determine the effect of cottonseed hull (CH) in the diets of rams on intake, digestibility, nitrogen balance, ingestive behaviour, and blood metabolites. Twenty males, uncastrated with an average body weight of 29.08 ± 4.18 kg, were distributed in a completely randomized design with four treatment diets and five replicates. The diet contained a forage to concentrate ratio of 50â:â50 (ground corn, soybean meal, corn silage, and CH), and the experimental treatments were inclusion of 0, 10, 20, and 30% CH (dry matter basis). Inclusion of CH linearly increased water intake, effective intake of ethereal extract and non-fibrous carbohydrates, and crude protein digestibility. Dry matter (DM) and neutral detergent fiber (NDF) digestibility, nitrogen faecal losses, and nitrogen balance reduced linearly with CH inclusion in the diet. Inclusion of CH in rams' diet reduces intake, nutrient digestibility, and intake and rumination efficiencies. Thus, it is not recommended to add this by-product to the diet of high-performance rams, but for nurture that does not aim at high performance, inclusion of up to 5% of this by-product is recommended.
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Aceite de Semillas de Algodón , Lactancia , Animales , Femenino , Masculino , Aceite de Semillas de Algodón/metabolismo , Dieta , Fibras de la Dieta/metabolismo , Digestión , Conducta Alimentaria , Nitrógeno/metabolismo , Rumen/metabolismo , Ovinos , Oveja Doméstica/metabolismo , Ensilaje , Zea mays/metabolismoRESUMEN
Fructooligosaccharides (FOS) are prebiotics of interest to the food industry. These compounds can be produced through the transfructosylation reaction by the enzyme fructofuranosidase. This enzyme is widely produced by fungi in a medium rich in sugar. Therefore, in this work, the main objectives were production, purification, biochemical characterization of a novel fructofuranosidase enzyme by Penicillium citreonigrum URM 4459 and synthesize and evaluate the antibacterial potential of fructooligosaccharides. With respect to sucrose hydrolysis, the optimal pH was 5.5, the apparent Km for purified FFase was 3.8 mM, the molecular mass was 43.0 kDa, estimated by gel filtration on Superdex increase G75 controlled by AKTA Avant 25 and confirmed by 10% SDS-PAGE under denaturing condition. Also, the isoelectric point was 4.9. The fractions obtained with enzymatic activities, both stable at acidic pH and high temperatures, as well as being able to produce FOS. Regarding antibacterial activity, the FOS produced in this study showed better results than commercial FOS and other carbon sources. Thus, this work presents relevant data for the use of P. citreonigum to produce fructofuranosidase and consequently FOS and can be used in the food and pharmaceutical industry.