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The development of an effective amphotericin B (AmB) formulation to replace actual treatments available for leishmaniasis, which present serious drawbacks, is a challenge. Here we report the development of hyaluronic acid-amphotericin B self-assembled nanocomplexes (HA-AmB), processed by freeze-drying (FD) or nano spray-drying (SD), using a simple process that favors the non-covalent drug-polysaccharide association in an amorphous state. These water-soluble formulations, which presented a nanometric size (300-600 nm), high colloidal stability (zeta potential around -39 mV) and an AmB loading (15-18%) in aggregated and super aggregated states, demonstrated less in vitro cytotoxic and hemolytic effects compared to the free-drug. A significant decrease in the number of intramacrophagic L. infantum amastigotes upon treatment (IC50 of 0.026 and 0.030 µM for HA-AmB FD and HA-AmB SD, respectively) was also observed, and the best selectivity index (SI) was observed for the HA-AmB SD nanocomplex (SI of 651). Intravenous administration of the HA-AmB SD nanocomplex for 3 alternate days showed an effective parasite reduction in the spleen and liver of C57BL/6 mice without signs of toxicity commonly observed upon free-AmB treatment. Although lower than that achieved with AmBisome® in the liver, the observed parasite reduction for the nanocomplex was of a similar order of magnitude. The efficacy, stability, safety and low cost of the HA-AmB SD nanocomplex highlight its potential as an alternative treatment for leishmaniasis.
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Anfotericina B , Leishmaniasis , Anfotericina B/farmacología , Animales , Sistemas de Liberación de Medicamentos , Ácido Hialurónico/uso terapéutico , Leishmaniasis/tratamiento farmacológico , Ratones , Ratones Endogámicos C57BLRESUMEN
Feeding-regulatory peptides such as thyrotropin-releasing hormone (TRH), α-melanocyte-stimulating hormone (α-MSH) and their receptors are expressed in brain regions involved in the homeostatic and hedonic control of food intake, such as the hypothalamus and the mesolimbic system, respectively. The nucleus accumbens (NAc) is part of the latter, a brain circuit involved in processing reward stimuli and the appetitive motivation of feeding. When TRH or α-MSH are administered in the NAc, both decrease food intake, through activating their respective receptors, TRH-R1 and MC4R. The actions of α-MSH as a homeostatic feeding-regulator involves the increase of hypothalamic TRH expression, thus, we aimed to identify whether TRH signaling in the NAc was also participating in α-MSH-induced reduction of food intake. α-MSH administration in the NAc of 48â¯h fasted rats reduced their food intake during the 2-h period of refeeding, increased accumbal TRH mRNA expression and decreased that of MC4R. Such downregulated MC4R mRNA levels implied a compensatory decrease of α-MSH actions in the NAc after the previous pathway stimulation. The co-administration of α-MSH along with an antisense oligonucleotide directed against pro-TRH mRNA in the NAc impaired the α-MSH-induced feeding reduction, supporting that the accumbal TRHergic pathway is downstream of α-MSH actions to inhibit feeding. Our results suggested that TRH in the NAc mediates some effects of α-MSH on inhibition of food intake; this supports the role of TRH not only as a homeostatic regulator but also as modulating the motivational aspects of feeding.
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Ingestión de Alimentos , Núcleo Accumbens/metabolismo , Precursores de Proteínas/metabolismo , Ácido Pirrolidona Carboxílico/análogos & derivados , Hormona Liberadora de Tirotropina/metabolismo , alfa-MSH/metabolismo , Animales , Masculino , Ácido Pirrolidona Carboxílico/metabolismo , ARN Mensajero/metabolismo , Ratas WistarRESUMEN
In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages (BMMΦ) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and 10 µg of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline Avicel-plus® CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. Avicel-plus® CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.
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In 2012, the first-generation protease inhibitors telaprevir (TVR) and boceprevir (BOC) were introduced in the Brazilian health system for treatment of chronic hepatitis C, after their approval by the National Committee for Health Technology Incorporation (CONITEC). However, these medicines were discontinued in 2015. The short period of use in therapy and their high cost require a discussion about the consequences for patients and for the health system of the early incorporation of new therapies. The article presents a qualitative analysis of the incorporation process of both medications in Brazil and the results of a multicenter study that included patients treated with BOC or TVR between January 2011 and December 2015 in five Brazilian cities. The study included 855 patients (BOC: n=247) and (TVR: n=608). The document analysis showed that CONITEC's decision to incorporate BOC and TVR was based on results of phase III clinical trials that compared sustained virologic response (SVR) rates of patients treated with BOC and TVR with rates of those that received placebo. However, these studies included a low percentage of cirrhotic patients. The SVR rates observed in this multicenter study were worse than clinical trials pointed out (BOC: 45.6%; TVR: 51.8%), but similar to those achieved with previously adopted therapies. The discontinuation rate due to adverse events was (BOC: 15.4%; TVR: 12.7%). Based on these unsatisfactory results, the study brings a discussion that goes beyond the therapy outcomes, exploring the incorporation of these high-cost medicines and the related decision-making process, contributing to future decisions in medicine policies and in the treatment of chronic hepatitis C.
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Antivirales/administración & dosificación , Hepatitis C Crónica/tratamiento farmacológico , Oligopéptidos/administración & dosificación , Prolina/análogos & derivados , Inhibidores de Proteasas/administración & dosificación , Antivirales/economía , Protocolos Clínicos , Toma de Decisiones , Quimioterapia Combinada , Femenino , Genotipo , Hepacivirus/genética , Hepatitis C Crónica/economía , Humanos , Interferón alfa-2 , Interferón-alfa/administración & dosificación , Masculino , Persona de Mediana Edad , Oligopéptidos/economía , Polietilenglicoles/administración & dosificación , Prolina/administración & dosificación , Prolina/economía , Inhibidores de Proteasas/economía , Proteínas Recombinantes/administración & dosificación , Estudios Retrospectivos , Ribavirina/administración & dosificaciónRESUMEN
We have, in previous work developed, characterized and evaluated the biocompatibility of an engineered hyaluronic acid nanogel. Here we assess the targetability of a hyaluronic acid nanogel towards CD44 overexpressing cells, in vitro and in vivo. Results obtained by flow cytometry and confocal fluorescence microscopy shows that nanogel is greatly internalized by non-small cancer lung cells (A549 cells), that overexpress CD44 receptors. The biodistribution and tumor targetability of the nanogel labelled with a near-infrared (NIR) probe were performed, in mice, through a non-invasive imaging system. Results revealed nanogel high targetability towards an induced subcutaneous A549 tumor. Nanogels pharmacokinetics was evaluated also in healthy animals, and Alexa Fluor 680 labelled nanogel exhibited higher accumulation in liver, kidneys and skin. Also, a comparative biodistribution study was performed, using two NIR imaging probes, Cy5.5 and Alexa Fluor 680.
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Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Geles , Ácido Hialurónico/administración & dosificación , Neoplasias Pulmonares/tratamiento farmacológico , Nanotecnología , Células A549 , Animales , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Humanos , Ácido Hialurónico/farmacocinética , Técnicas In Vitro , Riñón/metabolismo , Hígado/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Masculino , Ratones Desnudos , Piel/metabolismo , Espectroscopía Infrarroja Corta , Distribución TisularRESUMEN
Tuberculosis (TB), a disease caused by the human pathogen Mycobacterium tuberculosis, recently joined HIV/AIDS on the top rank of deadliest infectious diseases. Low patient compliance due to the expensive, long-lasting and multi-drug standard therapies often results in treatment failure and emergence of multi-drug resistant strains. In this scope, antimicrobial peptides (AMPs) arise as promising candidates for TB treatment. Here we describe the ability of the exogenous AMP LLKKK18 to efficiently kill mycobacteria. The peptide's potential was boosted by loading into self-assembling Hyaluronic Acid (HA) nanogels. These provide increased stability, reduced cytotoxicity and degradability, while potentiating peptide targeting to main sites of infection. The nanogels were effectively internalized by macrophages and the peptide presence and co-localization with mycobacteria within host cells was confirmed. This resulted in a significant reduction of the mycobacterial load in macrophages infected in vitro with the opportunistic M. avium or the pathogenic M. tuberculosis, an effect accompanied by lowered pro-inflammatory cytokine levels (IL-6 and TNF-α). Remarkably, intra-tracheal administration of peptide-loaded nanogels significantly reduced infection levels in mice infected with M. avium or M. tuberculosis, after just 5 or 10 every other day administrations. Considering the reported low probability of resistance acquisition, these findings suggest a great potential of LLKKK18-loaded nanogels for TB therapeutics.
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Péptidos Catiónicos Antimicrobianos/administración & dosificación , Antituberculosos/administración & dosificación , Portadores de Fármacos/administración & dosificación , Geles/administración & dosificación , Ácido Hialurónico/administración & dosificación , Nanoestructuras/administración & dosificación , Animales , Péptidos Catiónicos Antimicrobianos/química , Péptidos Catiónicos Antimicrobianos/uso terapéutico , Antituberculosos/química , Antituberculosos/uso terapéutico , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Portadores de Fármacos/química , Portadores de Fármacos/uso terapéutico , Geles/química , Geles/uso terapéutico , Ácido Hialurónico/química , Ácido Hialurónico/uso terapéutico , Interleucina-6/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/microbiología , Ratones Endogámicos C57BL , Mycobacterium avium/efectos de los fármacos , Mycobacterium avium/crecimiento & desarrollo , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/crecimiento & desarrollo , Nanoestructuras/química , Nanoestructuras/uso terapéutico , Tuberculosis Pulmonar/tratamiento farmacológico , Tuberculosis Pulmonar/microbiología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Biodegradable poly(l-lactic acid) (PLLA) and PLLA/CoFe2O4 magnetic microspheres with average sizes ranging between 0.16-3.9µm and 0.8-2.2µm, respectively, were obtained by an oil-in-water emulsion method using poly(vinyl alcohol) (PVA) solution as the emulsifier agent. The separation of the microspheres in different size ranges was then performed by centrifugation and the colloidal stability assessed at different pH values. Neat PLLA spheres are more stable in alkaline environments when compared to magnetic microspheres, both types being stable for pHs higher than 4, resulting in a colloidal suspension. On the other hand, in acidic environments the microspheres tend to form aggregates. The neat PLLA microspheres show a degree of crystallinity of 40% whereas the composite ones are nearly amorphous (17%). Finally, the biocompatibility was assessed by cell viability studies with MC3T3-E1 pre-osteoblast cells.
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Tecnología Biomédica , Fenómenos Magnéticos , Microesferas , Poliésteres/química , Poliésteres/aislamiento & purificación , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Concentración de Iones de Hidrógeno , Tamaño de la Partícula , Poliésteres/farmacología , Propiedades de SuperficieRESUMEN
A novel approach for tissue engineering applications based on the use of magnetoelectric materials is presented. This work proves that magnetoelectric Terfenol-D/poly(vinylidene fluoride-co-trifluoroethylene) composites are able to provide mechanical and electrical stimuli to MC3T3-E1 pre-osteoblast cells and that those stimuli can be remotely triggered by an applied magnetic field. Cell proliferation is enhanced up to ≈ 25% when cells are cultured under mechanical (up to 110 ppm) and electrical stimulation (up to 0.115 mV), showing that magnetoelectric cell stimulation is a novel and suitable approach for tissue engineering allowing magnetic, mechanical and electrical stimuli.
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Proliferación Celular , Campos Magnéticos , Osteoblastos/citología , Ingeniería de Tejidos/métodos , Aleaciones/química , Animales , Línea Celular , Disprosio/química , Estimulación Eléctrica , Hidrocarburos Fluorados/química , Hierro/química , Membranas Artificiales , Ratones , Microscopía Fluorescente , Reproducibilidad de los Resultados , Terbio/química , Ingeniería de Tejidos/instrumentación , Andamios del Tejido/química , Compuestos de Vinilo/químicaRESUMEN
The biodistribution profile of a new dextrin nanomagnetogel, which consists of γ-Fe2O3 superparamagnetic nanoparticles loaded within a polymeric matrix of modified dextrin, was studied in mice. The nanomagnetogel bear a monomodal size distribution profile (average diameter 110 nm) close to neutral surface charge and higher relaxivity (r2 = 215-248 mM(-1) s(-1) and r2/r1 = 13-11) than those of commercial formulations (r2 = 160-177 mM(-1) s(-1) and r2/r1 = 4-7). Also, the observed blood half-life-approximately 4 h-is superior to that of similar commercially available formulations, which remain for a few minutes in circulation. PEGylation resulted in 1.7- and 1.2-fold lower accumulation in the liver and spleen, respectively, within the first 24 h. Noteworthy, a good correlation was obtained between the amount of polymer (quantified by scintigraphy) in the spleen, 48 h after administration, and the amount of iron physically loaded through hydrophobic interactions (quantified by ICP) indicating the absence of iron leakage from the polymeric matrix. This study provides evidence of the in vivo stability of a self-assembled nanomagnetogel, a relevant feature which is seldom reported in the literature.
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Dextrinas/farmacocinética , Portadores de Fármacos , Compuestos Férricos/farmacocinética , Nanopartículas de Magnetita/química , Animales , Dextrinas/química , Estabilidad de Medicamentos , Compuestos Férricos/química , Geles , Semivida , Interacciones Hidrofóbicas e Hidrofílicas , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/ultraestructura , Imagen por Resonancia Magnética , Nanopartículas de Magnetita/ultraestructura , Ratones , Polietilenglicoles/química , Bazo/efectos de los fármacos , Bazo/metabolismo , Bazo/ultraestructura , Electricidad Estática , Propiedades de Superficie , Tomografía Computarizada de Emisión de Fotón ÚnicoRESUMEN
Poly(l-lactide) electrospun mats with random and aligned fiber orientation and films have been produced with degrees of crystallinity ranging from 0 up to nearly 50%. The overall surface roughness is practically constant irrespective of the sampling areas (1 × 1 µm to 20 × 20 µm) for degrees of crystallinity below 30%, increasing for higher degrees of crystallinity for the larger sampling areas. Further, due to fiber confinement, surface roughness variations are smaller in electrospun mats. Samples with 50% of crystallinity show the lowest osteoblast and the highest fibroblast proliferation. Therefore, it is verified that higher roughness promotes lower osteoblast but higher fibroblast proliferation. The overall results indicate the relevant role of the sub-microenvironment variations associated to the microscale roughness in determining the different cell responses.
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Fibroblastos/citología , Osteoblastos/citología , Poliésteres/química , Células 3T3 , Animales , Proliferación Celular , Cristalización , Ratones , Propiedades de SuperficieRESUMEN
In this work, polypyrrole (PPy) was in situ polymerized onto the surface of bacterial nanocellulose (BNC) produced by Gluconacetobacter xylinus, by chemical oxidation in aqueous medium using ammonium persulfate. Composites (BNC/PPy) were produced with varying concentrations of pyrrole (Py). The produced BNC/PPy membranes were used as a template for the seeding of PC12 rat neuronal cells. Cell suspensions were directly seeded onto the surfaces of the BNC/PPy membranes. The Py concentration affected the behavior of neuronal cells that adhered and grew significantly more on BNC/PPy comparatively to BNC. Scanning electron microscopy (SEM) micrographs revealed that PC12 cells adhered on the surface of the BNC and BNC/PPy membranes. Conductive PPy coatings on nanofibers acting as an active interface for tissue engineering may be used to regulate cell activity through electrical stimulations.
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Celulosa/química , Materiales Biocompatibles Revestidos/farmacología , Neuronas/efectos de los fármacos , Polímeros/química , Polímeros/farmacología , Pirroles/química , Pirroles/farmacología , Andamios del Tejido/química , Animales , Materiales Biocompatibles Revestidos/química , Conductividad Eléctrica , Ensayo de Materiales , Nanopartículas/química , Neuronas/fisiología , Células PC12 , Polisacáridos Bacterianos/química , Ratas , Propiedades de Superficie , Técnicas de Cultivo de Tejidos/instrumentación , Ingeniería de TejidosRESUMEN
This study aims at the production and characterization of a "nanomagnetogel" consisting of superparamagnetic iron oxide nanoparticles (γ-Fe2O3) stabilized within a hydrophobized-dextrin nanogel. The nanomagnetogel obtained was extensively characterized with respect to physico-chemical (transmission electron microscopy, cryo-scanning electron microscopy, dynamic light scattering, small angle X-ray scattering), magnetic (relaxometry, MIAplex) and biocompatibility (interaction with cells) properties. The obtained nanomagnetogel formulation, with about 4 mM of iron and a diameter of 100 nm, presents relevant features as a promising magnetic resonance imaging (MRI) contrast agent, noteworthy superparamagnetic behavior, high stability, narrow size distribution and potential for magnetic guidance to target areas by means of an external magnetic field. High values of transverse relaxivity make the nanomagnetogel a promising T2 contrast agent, allowing enhanced lesion detectability through magnetic resonance imaging. The nanomagnetogel demonstrated non-toxicity for 3T3 fibroblast cultures and was efficiently internalized by bone marrow-derived macrophages, therefore having potential as a contrast agent for MRI of the organs associated with the reticuloendothelial system (spleen, liver). The production of the nanomagnetogel is simple and easy to scale up, thus offering great technological potential.
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BACKGROUND AND AIM: Disturbances in intestinal motility have been described since XIX century, with a not very well understood pathogenesis and few therapeutic approaches. Considering chewing as an important stimulus to promote intestinal motility we designed this study to assess the efficacy of chewing gum to improve the postoperative ileus, and the clinical relevance of this intervention. METHODS: Were included patients who underwent for elective left hemicolectomy during January 2007 to December 2008. The patients were randomly assigned to receive chewing gum or nothing during the post-surgery period. The main outcomes assessed were duration of postoperative ileus, and reduction in hospitalization days. RESULTS: During the period of the study were included 32 patients, were included mainly men (11 and 9 in treated and control patients, respectively). The first defecation after surgery procedure occurs at (44.2 ± 17.6 h) in the treated group and (55.5 ± 14.8 h) in the control group (p = 0.05). In the same way oral tolerance was achieved faster in the treated group (p= 0.05). The hospital stay duration was not different among treated and not treated patients. CONCLUSIONS: The use of chewing gum in patients after elective left hemicolectomy, reduce the duration of the postoperative ileus. However, this does not reduce the hospital stay duration.
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Goma de Mascar , Colectomía , Ileus/terapia , Complicaciones Posoperatorias/terapia , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Tiempo de Internación , Masculino , Persona de Mediana EdadRESUMEN
The characterization of biodistribution is a central requirement in the development of biomedical applications based on the use of nanoparticles, in particular for controlled drug delivery. The blood circulation time, organ biodistribution and rate of excretion must be well characterized in the process of product development. In this work, the biodistribution of recently developed self-assembled dextrin nanoparticles is addressed. Functionalization of the dextrin nanoparticles with a DOTA-monoamide-type metal chelator, via click chemistry, is described. The metal chelator functionalized nanoparticles were labelled with a gamma-emitting (153)Sm(3+) radioisotope and the blood clearance rate and organ biodistribution of the nanoparticles were obtained. The effect of PEG surface coating on the blood clearance rate and organ biodistribution of the nanoparticles was also studied.
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Dextrinas/farmacocinética , Sistemas de Liberación de Medicamentos/métodos , Nanopartículas , Animales , Técnicas Químicas Combinatorias , Dextrinas/sangre , Dextrinas/química , Compuestos Heterocíclicos con 1 Anillo/química , Tamaño de la Partícula , Polietilenglicoles/química , Ratas , Ratas Wistar , Samario/química , Compuestos de Sulfhidrilo/química , Distribución TisularRESUMEN
The attachment of cells to biomedical materials can be improved by using adhesion sequences, such as Arg-Gly-Asp (RGD), found in several extracellular matrix proteins. In this work, bifunctional recombinant proteins, with a Cellulose-Binding Module (CBM), from the cellulosome of Clostridium thermocellum and cell binding sequences-RGD, GRGDY-were cloned and expressed in E.coli. These RGD-containing cellulose binding proteins were purified and used to coat bacterial cellulose fibres. Its effect on the cell adhesion/biocompatibility properties was tested using a mouse embryo fibroblasts culture. Bacterial cellulose (BC) secreted by Gluconacetobacter xylinus (=Acetobacter xylinum) is a material with unique properties and promising biomedical applications. CBMs adsorbs specifically and tightly on cellulose. Thus, they are a useful tool to address the fused RGD sequence (or other bioactive peptides) to the cellulose surface, in a specific and simple way. Indeed, fibroblasts exhibit improved ability to interact with bacterial cellulose sheets coated with RGD-CBM proteins, as compared with cellulose treated with the CBM, that is, without the adhesion peptide. The effect of the several fusion proteins produced was analyzed.
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Celulosa/metabolismo , Fibroblastos/citología , Oligopéptidos/metabolismo , Receptores de Superficie Celular/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Células 3T3 , Animales , Adhesión Celular , Supervivencia Celular/efectos de los fármacos , Clostridium thermocellum , Medios de Cultivo , Electroforesis en Gel de Poliacrilamida , Gluconacetobacter xylinus/química , Ratones , Microscopía Fluorescente , Poliestirenos/farmacología , Receptores de Superficie Celular/química , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/aislamiento & purificación , SueroRESUMEN
The electrophoretic profile of cerebrospinal fluid proteins and albumin quota was studied in healthy dogs and dogs with distemper in either nervous or non-nervous phases. Cerebrospinal fluid (CSF) samples from 30 dogs were collected by puncture of the cisterna magna. The total protein content, the albumin quota, and the electrophoretic fraction of CSF proteins in agarose gel plates were evaluated. Results were similar in healthy dogs and dogs with distemper and no nervous signs, but were significantly increased in the group of dogs with distemper showing nervous signs. The study of CSF protein profile proved useful and contributed significantly on the detection of central nervous system disorders and damages to the blood-brain barrier during the nervous phase of distemper.
Estudaram-se o perfil eletroforético das proteínas liquóricas e a cota de albumina em cães sem e com cinomose na fase neurológica e não-neurológica. A punção da cisterna magna para a obtenção de amostras de liquor realizou-se em 30 cães. Analisaram-se teores de proteínas totais, cota de albumina e fracionamento eletroforético das proteínas liquóricas em gel de agarose. Os resultados foram semelhantes nos cães normais e nos cães com cinomose sem sinais neurológicos e significativamente elevados no grupo de cães com cinomose apresentando sinais neurológicos. O estudo do quadro protéico do líquido cérebroespinhal foi útil e contribuiu significativamente na detecção de lesões ao sistema nervoso central e de danos à barreira hematoencefálica durante a fase neurológica da cinomose.
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Animales , Masculino , Femenino , Moquillo/diagnóstico , Moquillo/líquido cefalorraquídeo , Perros , Electroforesis/métodos , Líquido Cefalorraquídeo/fisiologíaRESUMEN
Enzymatic versus chemical deinking is examined for MOW and photocopy prints. Several enzymatic preparations and two fibre/ink particle separation methods are tested. Deinking was monitored by image analysis and standard pulp and paper characterisation procedures. The effectiveness of the fibre/ink particle separation method depends on the ink particle's size: for smaller particles a washing step is recommended whereas for larger particles, the use of flotation is necessary. The enzymatic treatment is a competitive alternative for MOW and photocopy paper deinking. However, the process requires the selection of an adequate enzymatic preparation for each paper grade.
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Conservación de los Recursos Naturales , Tinta , Modelos Biológicos , Biodegradación Ambiental , Celulasa/química , Estabilidad de Enzimas , Procesamiento de Imagen Asistido por Computador , Papel , Tamaño de la Partícula , Impresión , Eliminación de ResiduosRESUMEN
Two enzymatic extracts obtained from xylan-grown Aspergillus terreus CCMI 498 and cellulose-grown Trichoderma viride CCMI 84 were characterised for different glycanase activities. Both strains produce extracellular endoxylanase and endoglucanase enzymes. The enzymes optimal activity was found in the temperature range of 45-60 degrees C. Endoglucanase systems show identical activity profiles towards temperature, regardless of the strain and inducing substrate. Conversely, the endoxylanases produced by both strains showed maximal activity at different pH values (from 4.5 to 5.5), being the more acidic xylanase produced by T. viride grown on cellulose. The endoglucanase activities have an optimum pH at 4.5-5.0. The endoxylanase and endoglucanase activities exhibited high stability at 50 degrees C and pH 5.0. Mannanase, beta-xylosidase, and amylase activities were also found, being the first two activities only present for T. viride extract. These two enzymatic extracts were used for mixed office wastepaper (MOW) deinking. When the enzymatic extract from T. viride was used, a further increase of 24% in ink removal was obtained by comparison with the control. Both enzymes contributed to the improvement of the paper strength properties and the obtained results clearly indicate that the effective use of enzymes for deinking can also contribute to the pulp and paper properties improvement.
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Celulasa/química , Tinta , Papel , Eliminación de Residuos/métodos , Xilosidasas/química , Aspergillus/química , Aspergillus/enzimología , Biodegradación Ambiental , Sistema Libre de Células , Células Cultivadas , Celulasa/aislamiento & purificación , Celulasa/metabolismo , Endo-1,4-beta Xilanasas , Activación Enzimática , Estabilidad de Enzimas , Tamaño de la Partícula , Protones , Sensibilidad y Especificidad , Especificidad de la Especie , Temperatura , Trichoderma/química , Trichoderma/enzimología , Xilosidasas/aislamiento & purificación , Xilosidasas/metabolismoRESUMEN
A cellulase from Trichoderma reesei was immobilized on Eudragit L-100, a reversibly soluble polymer depending on the pH of the medium. The solubility of the modified cellulase was studied at different pH values. By changing the pH, the adsorption equilibrium of the derivatized proteins is switched towards the liquid phase, thus making recycling possible. This method allows for improved stability, without major loss of specific activity. The adsorption of cellulase on Eudragit lowers the enthalpy of denaturation, but affects only slightly the denaturation temperature. The use of carbodiimide was ineffective on linking the enzymes covalently to the polymer, since the immobilization process was found to be only mediated by non-covalent forces.
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Celulasa/metabolismo , Ácidos Polimetacrílicos/química , Ácidos Polimetacrílicos/metabolismo , Adsorción , Estabilidad de Enzimas , Enzimas Inmovilizadas , Concentración de Iones de Hidrógeno , Sensibilidad y Especificidad , Solubilidad , Trichoderma/enzimologíaRESUMEN
The selective extraction of capsaicinoids and carotenoids from chili guajillo "puya" flour was studied. When ethanol was used as solvent, 80% of capsaicinoids and 73% of carotenoids were extracted, representing an interesting alternative for the substitution of hexane in industrial processes. Additionally, when the flour was pretreated with enzymes that break the cell wall and then dried, extraction in ethanol increased to 11 and 7% for carotenoid and capsaicinoid, respectively. A selective two-stage extraction process after the treatment with enzymes is proposed. The first step uses 30% (v/v) ethanol and releases up to 60% of the initial capsaicinoids, and the second extraction step with industrial ethanol permits the recovery of 83% of carotenoids present in the flour.