RESUMEN
Bdellovibrios are predatory bacteria that invade other live Gram-negative bacterial cells for growth and reproduction. They have recently been considered as potential living antibiotics and biocontrol agents. In this study, the predatory activity and biocontrol potency of Bdellovibrio bacteriovorus strain SOIR-1 against Pantoea sp. strain BCCS and Xanthomonas campestris, two exo-biopolymer-producing phytopathogens, was evaluated. Plaque formation assays and lysis analysis in the broth co-cultures were used for the in vitro evaluation of bacteriolytic activity of strain SOIR-1. The in vivo biocontrol potential of strain SOIR-1 was evaluated by pathogenicity tests on the onion bulbs and potato tuber slices. The phytopathogens were also recovered from the infected plant tissues and confirmed using biochemical tests and PCR-based 16S rRNA gene sequence analysis. Typical bdellovibrios plaques were developed on the lawn cultures of Pantoea sp. BCCS and X. campestris. The killing rate of strain SOIR-1 toward Pantoea sp. BCCS and X. campestris was 84.3% and 76.3%, respectively. Exo-biopolymers attenuated the predation efficiency of strain SOIR-1 up to 10.2-18.2% (Pantoea sp. BCCS) and 12.2-17.3% (X. campestris). The strain SOIR-1 significantly reduced rotting symptoms in the onion bulbs caused by Pantoea sp. BCCS (69.0%) and potato tuber slices caused by X. campestris (73.1%). Although more field assessments are necessary, strain SOIR-1 has the preliminary potential as a biocontrol agent against phytopathogenic Pantoea sp. BCCS and X. campestris, especially in postharvest storage. Due to the particular physicochemical properties of evaluated exo-biopolymers, they can be used in the designing encapsulation systems for delivery of bdellovibrios.
Asunto(s)
Bdellovibrio bacteriovorus/fisiología , Bdellovibrio bacteriovorus/patogenicidad , Agentes de Control Biológico/farmacología , Pantoea/efectos de los fármacos , Pantoea/fisiología , Xanthomonas campestris/efectos de los fármacos , Xanthomonas campestris/fisiología , Antibiosis , Biopolímeros/fisiología , Técnicas de Cocultivo/métodos , ADN Bacteriano , Interacciones Microbianas , ARN Ribosómico 16SRESUMEN
The slug Arion subfuscus produces a mucus-based defensive secretion that is remarkably tough. This glue appears to be a double network hydrogel, gaining its toughness through the synergistic actions of two networks of polymers, a relatively stiff network and a relatively deformable network. The double network mechanism has great potential to guide the development of synthetic adhesives. Mechanical tests were performed to analyse key predictions of the mechanism. Stress relaxation tests and tensile tests support the presence of stable cross-links. Cyclic stress-strain tests demonstrate that the glue dissipates a great deal of energy through the failure of these cross-links as sacrificial bonds. Energy dissipation by failure of sacrificial bonds rather than viscous processes is supported by the minimal effect of the time course of the experiments on the measured properties. These sacrificial bonds appear able to reform within minutes after failure. Finally, the glue's stiffness decreases at pH values below 5.5, whereas magnesium and calcium rapidly dissociate from the glue at all pH values tested. Thus, these ions may not be the primary cross-linkers generating the glue's stiffness. This article is part of the theme issue 'Transdisciplinary approaches to the study of adhesion and adhesives in biological systems'.
Asunto(s)
Biopolímeros/fisiología , Elasticidad , Metabolismo Energético , Gastrópodos/fisiología , Animales , HidrogelesRESUMEN
The mechanical properties of the extracellular matrix (ECM)-a complex, 3D, fibrillar scaffold of cells in physiological environments-modulate cell behavior and can drive tissue morphogenesis, regeneration, and disease progression. For simplicity, it is often convenient to assume these properties to be time-invariant. In living systems, however, cells dynamically remodel the ECM and create time-dependent local microenvironments. Here, we show how cell-generated contractile forces produce substantial irreversible changes to the density and architecture of physiologically relevant ECMs-collagen I and fibrin-in a matter of minutes. We measure the 3D deformation profiles of the ECM surrounding cancer and endothelial cells during stages when force generation is active or inactive. We further correlate these ECM measurements to both discrete fiber simulations that incorporate fiber crosslink unbinding kinetics and continuum-scale simulations that account for viscoplastic and damage features. Our findings further confirm that plasticity, as a mechanical law to capture remodeling in these networks, is fundamentally tied to material damage via force-driven unbinding of fiber crosslinks. These results characterize in a multiscale manner the dynamic nature of the mechanical environment of physiologically mimicking cell-in-gel systems.
Asunto(s)
Matriz Extracelular/fisiología , Seudópodos/fisiología , Fenómenos Biomecánicos , Biopolímeros/química , Biopolímeros/fisiología , Línea Celular , Microambiente Celular/fisiología , Biología Computacional , Simulación por Computador , Matriz Extracelular/química , Matriz Extracelular/ultraestructura , Células Endoteliales de la Vena Umbilical Humana , Humanos , Imagenología Tridimensional , Cinética , Modelos Biológicos , Seudópodos/química , Seudópodos/ultraestructuraRESUMEN
The chemical functionalities within biopolymers determine their physical properties and biological activities. The relationship between the side chains available to a biopolymer population and the potential functions of the resulting polymers, however, has proven difficult to study experimentally. Using seven sets of chemically diverse charged, polar, and nonpolar side chains, we performed cycles of artificial translation, in vitro selections for binding to either PCSK9 or IL-6 protein, and replication on libraries of random side chain-functionalized nucleic acid polymers. Polymer sequence convergence, bulk population target binding, affinity of individual polymers, and head-to-head competition among post-selection libraries collectively indicate that polymer libraries with nonpolar side chains outperformed libraries lacking these side chains. The presence of nonpolar groups, resembling functionality existing in proteins but missing from natural nucleic acids, thus may be strong determinants of binding activity. This factor may contribute to the apparent evolutionary advantage of proteins over their nucleic acid precursors for some molecular recognition tasks.
Asunto(s)
Biopolímeros/química , Biopolímeros/fisiología , Replicación del ADN , Humanos , Interleucina-6/química , Biblioteca de Péptidos , Polímeros/química , Proproteína Convertasa 9/química , Proteínas/químicaRESUMEN
Life as we know it requires three basic types of polymers: polypeptide, polynucleotide, and polysaccharide. Here we evaluate both universal and idiosyncratic characteristics of these biopolymers. We incorporate this information into a model that explains much about their origins, selection, and early evolution. We observe that all three biopolymer types are pre-organized, conditionally self-complementary, chemically unstable in aqueous media yet persistent because of kinetic trapping, with chiral monomers and directional chains. All three biopolymers are synthesized by dehydration reactions that are catalyzed by molecular motors driven by hydrolysis of phosphorylated nucleosides. All three biopolymers can access specific states that protect against hydrolysis. These protected states are folded, using self-complementary interactions among recurrent folding elements within a given biopolymer, or assembled, in associations between the same or different biopolymer types. Self-association in a hydrolytic environment achieves self-preservation. Heterogeneous association achieves partner-preservation. These universal properties support a model in which life's polymers emerged simultaneously and co-evolved in a common hydrolytic milieu where molecular persistence depended on folding and assembly. We believe that an understanding of the structure, function, and origins of any given type of biopolymer requires the context of other biopolymers.
Asunto(s)
Biopolímeros/biosíntesis , Biopolímeros/metabolismo , Biopolímeros/fisiología , Animales , Catálisis , Humanos , Péptidos/metabolismo , Péptidos/fisiología , Polímeros , Polinucleótidos/biosíntesis , Polinucleótidos/metabolismo , Polisacáridos/biosíntesis , Polisacáridos/metabolismo , Polisacáridos/fisiología , Pliegue de Proteína , Pliegue del ARN/fisiologíaRESUMEN
How cells move through the three-dimensional extracellular matrix (ECM) is of increasing interest in attempts to understand important biological processes such as cancer metastasis. Just as in motion on flat surfaces, it is expected that experimental measurements of cell-generated forces will provide valuable information for uncovering the mechanisms of cell migration. However, the recovery of forces in fibrous biopolymer networks may suffer from large errors. Here, within the framework of lattice-based models, we explore possible issues in force recovery by solving the inverse problem: how can one determine the forces cells exert to their surroundings from the deformation of the ECM? Our results indicate that irregular cell traction patterns, the uncertainty of local fiber stiffness, the non-affine nature of ECM deformations and inadequate knowledge of network topology will all prevent the precise force determination. At the end, we discuss possible ways of overcoming these difficulties.
Asunto(s)
Biopolímeros/fisiología , Movimiento Celular , Matriz Extracelular/fisiología , Modelos BiológicosRESUMEN
We describe a multiscale model that incorporates force-dependent mechanical plasticity induced by interfiber cross-link breakage and stiffness-dependent cellular contractility to predict focal adhesion (FA) growth and mechanosensing in fibrous extracellular matrices (ECMs). The model predicts that FA size depends on both the stiffness of ECM and the density of ligands available to form adhesions. Although these two quantities are independent in commonly used hydrogels, contractile cells break cross-links in soft fibrous matrices leading to recruitment of fibers, which increases the ligand density in the vicinity of cells. Consequently, although the size of focal adhesions increases with ECM stiffness in nonfibrous and elastic hydrogels, plasticity of fibrous networks leads to a departure from the well-described positive correlation between stiffness and FA size. We predict a phase diagram that describes nonmonotonic behavior of FA in the space spanned by ECM stiffness and recruitment index, which describes the ability of cells to break cross-links and recruit fibers. The predicted decrease in FA size with increasing ECM stiffness is in excellent agreement with recent observations of cell spreading on electrospun fiber networks with tunable cross-link strengths and mechanics. Our model provides a framework to analyze cell mechanosensing in nonlinear and inelastic ECMs.
Asunto(s)
Matriz Extracelular/fisiología , Adhesiones Focales/fisiología , Modelos Biológicos , Actomiosina/química , Actomiosina/fisiología , Fenómenos Biofísicos , Biopolímeros/química , Biopolímeros/fisiología , Simulación por Computador , Módulo de Elasticidad , Matriz Extracelular/química , Adhesiones Focales/química , Humanos , Hidrogeles , Mecanotransducción Celular/fisiología , Fibras de Estrés/química , Fibras de Estrés/fisiologíaRESUMEN
The chemical compositions of the surface conditioning layers formed by different types of solutions (from isolated EPS to whole culture media), involving different bacterial strains relevant for biocorrosion were compared, as they may influence the initial step in biofilm formation. Different substrata (polystyrene, glass, steel) were conditioned and analyzed by X-ray photoelectron spectroscopy. Peak decomposition and assignment were validated by correlations between independent spectral data and the ubiquitous presence of organic contaminants on inorganic substrata was taken into account. Proteins or peptides were found to be a major constituent of all conditioning layers and polysaccharides were not present in appreciable concentrations; the proportion of nitrogen which may be due to DNA was lower than 15%. There was no significant difference between the compositions of the adlayers formed from different conditioning solutions, except for the adlayers produced with tightly bound EPS extracted from D. alaskensis.
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Proteínas Bacterianas , Biopelículas/crecimiento & desarrollo , Incrustaciones Biológicas/prevención & control , Biopolímeros , Espacio Extracelular , Proteínas Bacterianas/aislamiento & purificación , Proteínas Bacterianas/fisiología , Biopolímeros/aislamiento & purificación , Biopolímeros/fisiología , Medios de Cultivo/química , Espacio Extracelular/química , Espacio Extracelular/fisiología , Nitrógeno/análisis , Espectroscopía de Fotoelectrones/métodos , Polisacáridos/aislamiento & purificación , Propiedades de SuperficieRESUMEN
We have reported previously that a missense mutation in the mitochondrial fission gene Dynamin-related protein 1 (Drp1) underlies the Python mouse model of monogenic dilated cardiomyopathy. The aim of this study was to investigate the consequences of the C452F mutation on Drp1 protein function and to define the cellular sequelae leading to heart failure in the Python monogenic dilated cardiomyopathy model. We found that the C452F mutation increased Drp1 GTPase activity. The mutation also conferred resistance to oligomer disassembly by guanine nucleotides and high ionic strength solutions. In a mouse embryonic fibroblast model, Drp1 C452F cells exhibited abnormal mitochondrial morphology and defective mitophagy. Mitochondria in C452F mouse embryonic fibroblasts were depolarized and had reduced calcium uptake with impaired ATP production by oxidative phosphorylation. In the Python heart, we found a corresponding progressive decline in oxidative phosphorylation with age and activation of sterile inflammation. As a corollary, enhancing autophagy by exposure to a prolonged low-protein diet improved cardiac function in Python mice. In conclusion, failure of Drp1 disassembly impairs mitophagy, leading to a downstream cascade of mitochondrial depolarization, aberrant calcium handling, impaired ATP synthesis, and activation of sterile myocardial inflammation, resulting in heart failure.
Asunto(s)
Biopolímeros/fisiología , Dinaminas/fisiología , Insuficiencia Cardíaca/etiología , Mitofagia , Miocarditis/etiología , Animales , Biopolímeros/genética , Biopolímeros/metabolismo , Células Cultivadas , Dinaminas/genética , Dinaminas/metabolismo , Insuficiencia Cardíaca/fisiopatología , Ratones , Mutación , Miocarditis/fisiopatología , Fosforilación OxidativaRESUMEN
OBJETIVO: Determinar la sobrevida de pacientes con diagnóstico de cáncer gástrico en 2009-2010 en el Perú. MÉTODOS: Se realizó un estudio de tipo cohorte retrospectivo de pacientes con diagnóstico de cáncer gástrico registrados en el Sistema Nacional de Vigilancia Epidemiológica (SNVE) de la Dirección General de Epidemiología (DGE) y del Registro de Hechos Vitales (RHV) de la Oficina General de Estadística e Informática (OGEI) para los años 2009-2010. RESULTADOS: Se incluyeron 3 568 pacientes del SNVE, 51,5% eran hombres y 48,5% eran mujeres; la media de edad fue 63,9 años, 60,07% tenían 60 años o más. Se halló que 33,6% tenía adenocarcinoma de tipo intestinal, 18,7% tenía carcinoma de tipo difuso y 4,1% tenía linfoma gástrico primario. La sobrevida global fue de 29,7 ± 0,8 meses y fue mejor para los menores de 60 años (P = 0,034), para las mujeres (P = 0,014) y para el adenocarcinoma de tipo intestinal (P < 0,001). No hubo diferencias (P = 0,713) entre la sobrevida de los pacientes con linfoma gástrico y aquellos con adenocarcinoma. Para evaluar la tasa de mortalidad se incluyeron 6 069 registros de pacientes del RHV, la tasa de mortalidad nacional fue de 10,3 por cada 100 000 habitantes y las regiones con mayor mortalidad fueron Huánuco, Huancavelica y Junín. CONCLUSIONES: La sobrevida general fue de 29,7 ± 0,8 meses, las mujeres, los menores de 60 años y los pacientes con adenocarcinoma de tipo intestinal tienen mejor sobrevida. La mayor mortalidad por cáncer gástrico se concentra en las regiones más pobres del Perú, donde es probable que las condiciones de vida faciliten la alta transmisibilidad de Helicobacter pylori.
OBJECTIVE: Determine the survival rate of patients diagnosed with stomach cancer in 2009-2010 in Peru. METHODS: A retrospective cohort study was conducted of patients diagnosed with stomach cancer registered in the National Epidemiological Surveillance System (SNVE) of the Directorate General of Epidemiology (DGE) and the Register of Vital Statistics (RHV) of the General Office of Statistics and Information (OGEI) for the years 2009-2010. RESULTS: 3 568 patients of the SNVE were included; 51.5% were men and 48.5% were women; the average age was 63.9 years; 60.07% were 60 years old or older. It was found that 33.6% had intestinal type adenocarcinoma, 18.7% had diffuse type carcinoma, and 4.1% had primary gastric lymphoma. The overall survival rate was 29.7 ± 0.8 months and was better for those under 60 years (P = 0.034), for women (P = 0.014) and for intestinal type adenocarcinoma (P< 0.001). There was no difference (P = 0.713) between the survival rate of gastric lymphomas and adenocarcinomas. In order to evaluate mortality, 6 069 patient records from the RHV were included; national mortality was 10.3 per 100 000 population; the regions with the highest mortality were Huánuco, Huancavelica, and Junín. CONCLUSIONS: The general survival rate was 29.7 ± 0.8 months; women, those under 60 years, and patients with intestinal type adenocarcinoma had better survival rates. The highest mortality from stomach cancer is concentrated in the poorest regions of Peru, where it is probable that living conditions facilitate the high communicability of Helicobacter pylori.
Asunto(s)
Animales , Ratas , Péptidos beta-Amiloides/fisiología , Astrocitos/citología , Biopolímeros/fisiología , Neuronas/citología , Astrocitos/enzimología , Astrocitos/metabolismo , Calcio/metabolismo , /metabolismo , Activación Enzimática , Neuronas/enzimología , Neuronas/metabolismo , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismoRESUMEN
α-Synuclein (αSyn) aggregation and mitochondrial dysfunction both contribute to the pathogenesis of Parkinson disease (PD). Although recent studies have suggested that mitochondrial association of αSyn may disrupt mitochondrial function, it is unclear what aggregation state of αSyn is most damaging to mitochondria and what conditions promote or inhibit the effect of toxic αSyn species. Because the neuronal populations most vulnerable in PD are characterized by large cytosolic Ca(2+) oscillations that burden mitochondria, we examined mitochondrial Ca(2+) stress in an in vitro system comprising isolated mitochondria and purified recombinant human αSyn in various aggregation states. Using fluorimetry to simultaneously measure four mitochondrial parameters, we observed that soluble, prefibrillar αSyn oligomers, but not monomeric or fibrillar αSyn, decreased the retention time of exogenously added Ca(2+), promoted Ca(2+)-induced mitochondrial swelling and depolarization, and accelerated cytochrome c release. Inhibition of the permeability transition pore rescued these αSyn-induced changes in mitochondrial parameters. Interestingly, the mitotoxic effects of αSyn were specifically dependent upon both electron flow through complex I and mitochondrial uptake of exogenous Ca(2+). Our results suggest that soluble prefibrillar αSyn oligomers recapitulate several mitochondrial phenotypes previously observed in animal and cell models of PD: complex I dysfunction, altered membrane potential, disrupted Ca(2+) homeostasis, and enhanced cytochrome c release. These data reveal how the association of oligomeric αSyn with mitochondria can be detrimental to the function of cells with high Ca(2+)-handling requirements.
Asunto(s)
Biopolímeros/fisiología , Calcio/fisiología , Complejo I de Transporte de Electrón/metabolismo , Mitocondrias/fisiología , alfa-Sinucleína/fisiología , Animales , Benzotiazoles , Biopolímeros/química , Humanos , Potencial de la Membrana Mitocondrial , Ratones , Tiazoles/metabolismo , alfa-Sinucleína/químicaRESUMEN
Oligomers of the amyloid-ß (Aß) peptide have been implicated in the neurotoxicity associated with Alzheimer's disease. We have used single-molecule techniques to examine quantitatively the cellular effects of adding well characterized Aß oligomers to primary hippocampal cells and hence determine the initial pathway of damage. We found that even picomolar concentrations of Aß (1-40) and Aß (1-42) oligomers can, within minutes of addition, increase the levels of intracellular calcium in astrocytes but not in neurons, and this effect is saturated at a concentration of about 10 nM of oligomers. Both Aß (1-40) and Aß (1-42) oligomers have comparable effects. The rise in intracellular calcium is followed by an increase in the rate of ROS production by NADPH oxidase in both neurons and astrocytes. The increase in ROS production then triggers caspase-3 activation resulting in the inhibition of long-term potentiation. Our quantitative approach also reveals that only a small fraction of the oligomers are damaging and that an individual rare oligomer binding to an astrocyte can initiate the aforementioned cascade of responses, making it unlikely to be due to any specific interaction. Preincubating the Aß oligomers with an extracellular chaperone, clusterin, sequesters the oligomers in long-lived complexes and inhibits all of the physiological damage, even at a ratio of 100:1, total Aß to clusterin. To explain how Aß oligomers are so damaging but that it takes decades to develop Alzheimer's disease, we suggest a model for disease progression where small amounts of neuronal damage from individual unsequestered oligomers can accumulate over time leading to widespread tissue-level dysfunction.
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Péptidos beta-Amiloides/fisiología , Astrocitos/citología , Biopolímeros/fisiología , Neuronas/citología , Animales , Astrocitos/enzimología , Astrocitos/metabolismo , Calcio/metabolismo , Caspasa 3/metabolismo , Activación Enzimática , Neuronas/enzimología , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Small copy numbers of many molecular species in biological cells require stochastic models of the chemical reactions between the molecules and their motion. Important reactions often take place on one-dimensional structures embedded in three dimensions with molecules migrating between the dimensions. Examples of polymer structures in cells are DNA, microtubules, and actin filaments. An algorithm for simulation of such systems is developed at a mesoscopic level of approximation. An arbitrarily shaped polymer is coupled to a background Cartesian mesh in three dimensions. The realization of the system is made with a stochastic simulation algorithm in the spirit of Gillespie. The method is applied to model problems for verification and two more detailed models of transcription factor interaction with the DNA.
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Biopolímeros/fisiología , Modelos Biológicos , Modelos EstadísticosRESUMEN
The non-specific lipid transfer proteins (nsLTP) are unique to land plants. The nsLTPs are characterized by a compact structure with a central hydrophobic cavity and can be classified to different types based on sequence similarity, intron position or spacing between the cysteine residues. The type G nsLTPs (LTPGs) have a GPI-anchor in the C-terminal region which attaches the protein to the exterior side of the plasma membrane. The function of these proteins, which are encoded by large gene families, has not been systematically investigated so far. In this study we have explored microarray data to investigate the expression pattern of the LTPGs in Arabidopsis and rice. We identified that the LTPG genes in each plant can be arranged in three expression modules with significant coexpression within the modules. According to expression patterns and module sizes, the Arabidopsis module AtI is functionally equivalent to the rice module OsI, AtII corresponds to OsII and AtIII is functionally comparable to OsIII. Starting from modules AtI, AtII and AtIII we generated extended networks with Arabidopsis genes coexpressed with the modules. Gene ontology analyses of the obtained networks suggest roles for LTPGs in the synthesis or deposition of cuticular waxes, suberin and sporopollenin. The AtI-module is primarily involved with cuticular wax, the AtII-module with suberin and the AtIII-module with sporopollenin. Further transcript analysis revealed that several transcript forms exist for several of the LTPG genes in both Arabidopsis and rice. The data suggests that the GPI-anchor attachment and localization of LTPGs may be controlled to some extent by alternative splicing.
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Biopolímeros/metabolismo , Carotenoides/metabolismo , Lípidos/fisiología , Ceras/metabolismo , Empalme Alternativo/fisiología , Arabidopsis/metabolismo , Arabidopsis/fisiología , Biopolímeros/fisiología , Carotenoides/fisiología , Proteínas Portadoras , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/fisiología , Glicosilfosfatidilinositoles/metabolismo , Oryza/metabolismo , Oryza/fisiología , Filogenia , ARN de Planta/metabolismoRESUMEN
The copolymerization of poly(3-hydroxybutyrate) (PHB) is a promising trend in bioengineering to improve biomedical properties, e.g. biocompatibility, of this biodegradable polymer. We used strain Azotobacter chroococcum 7B, an effective producer of PHB, for biosynthesis of not only homopolymer and its main copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-HV), but also novel terpolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-poly(ethylene glycol) (PHB-HV-PEG), using sucrose as the primary carbon source and valeric acid and poly(ethylene glycol) 300 (PEG 300) as additional carbon sources. The chemical structure of PHB-HV-PEG was confirmed by (1)H nuclear-magnetic resonance analysis. The physico-chemical properties (molecular weight, crystallinity, hydrophilicity, surface energy) of produced biopolymer, the protein adsorption to the terpolymer, and cell growth on biopolymer films were studied. Despite of low EG-monomers content in bacterial-origin PHB-HV-PEG polymer, the terpolymer demonstrated significant improvement in biocompatibility in vitro in contrast to PHB and PHB-HV polymers, which may be coupled with increased protein adsorption, hydrophilicity and surface roughness of PEG-containing copolymer.
Asunto(s)
Azotobacter/metabolismo , Biopolímeros/fisiología , Adhesión Celular , Materiales Biocompatibles , Biopolímeros/biosíntesis , Rastreo Diferencial de Calorimetría , Proliferación Celular , Espectroscopía de Resonancia Magnética , Microscopía de Fuerza Atómica , Peso MolecularRESUMEN
We consider the effective surface motion of a particle that intermittently unbinds from a planar surface and performs bulk excursions. Based on a random-walk approach, we derive the diffusion equations for surface and bulk diffusion including the surface-bulk coupling. From these exact dynamic equations, we analytically obtain the propagator of the effective surface motion. This approach allows us to deduce a superdiffusive, Cauchy-type behavior on the surface, together with exact cutoffs limiting the Cauchy form. Moreover, we study the long-time dynamics for the surface motion.
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Biofisica/métodos , Biopolímeros/química , Algoritmos , Biopolímeros/fisiología , Difusión , Análisis de Fourier , Espectroscopía de Resonancia Magnética/métodos , Modelos Químicos , Modelos Estadísticos , Movimiento (Física) , Soluciones , Propiedades de SuperficieRESUMEN
Casparian strips are ring-like cell-wall modifications in the root endodermis of vascular plants. Their presence generates a paracellular barrier, analogous to animal tight junctions, that is thought to be crucial for selective nutrient uptake, exclusion of pathogens, and many other processes. Despite their importance, the chemical nature of Casparian strips has remained a matter of debate, confounding further molecular analysis. Suberin, lignin, lignin-like polymers, or both, have been claimed to make up Casparian strips. Here we show that, in Arabidopsis, suberin is produced much too late to take part in Casparian strip formation. In addition, we have generated plants devoid of any detectable suberin, which still establish functional Casparian strips. In contrast, manipulating lignin biosynthesis abrogates Casparian strip formation. Finally, monolignol feeding and lignin-specific chemical analysis indicates the presence of archetypal lignin in Casparian strips. Our findings establish the chemical nature of the primary root-diffusion barrier in Arabidopsis and enable a mechanistic dissection of the formation of Casparian strips, which are an independent way of generating tight junctions in eukaryotes.
Asunto(s)
Arabidopsis/fisiología , Biopolímeros/fisiología , Lignina/fisiología , Lípidos/fisiologíaRESUMEN
Filaments under distributed loads are common in biological systems. In this paper, we study the thermo-mechanical properties of an extensible thermally fluctuating elastic filament under distributed forces. The ground state of the filament is solved first, followed by an investigation of the thermal fluctuations around the ground state. We first consider a special case where the tangential component of the distributed force tau is uniform along the filament. For the force-extension relation in this case, we show that the filament is equivalent to one under end-to-end applied force F = tauL0/2 where L0 is the length of the filament. To study the thermal fluctuations under more general distributed loadings, the filament is first discretized into segments, and its energy is approximated up to quadratic order. Then the partition function of the discretized filament, or chain, is evaluated using multi-dimensional Gaussian integrals, from which free energy and other properties of the filament are derived. We show that a filament under distributed loads suffers larger thermal fluctuations than one with the end loads of the same magnitude. We also show that our results for a discretized filament agree with continuum theory for a continuous rod. Finally, we give some applications of our ideas to the stretching and fluctuation of DNA in non-uniform microfluidic channels.
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Fenómenos Biomecánicos/fisiología , Biopolímeros/fisiología , Citoesqueleto/fisiología , Actinas , ADN , Elasticidad , Análisis de Fourier , Microfluídica/métodos , Modelos Biológicos , TermodinámicaRESUMEN
The microtubule-associated protein tau exists as six isoforms created through the splicing of the second, third, and tenth exons. The isoforms are classified by their number of N-terminal exons (0N, 1N, or 2N) and by their number of microtubule-binding repeat regions (3R or 4R). Hyperphosphorylated isoforms accumulate in insoluble aggregates in Alzheimer's disease and other tauopathies. These neurodegenerative diseases can be categorized based on the isoform content of the aggregates they contain. Hyperphosphorylated tau has the general characteristics of an upward electrophoretic shift, decreased microtubule binding, and an association with aggregation. Previously we have shown that a combination of seven pseudophosphorylation mutations at sites phosphorylated by GSK-3ß, referred to as 7-Phos, induced several of these characteristics in full-length 2N4R tau and led to the formation of fewer but longer filaments. We sought to determine whether the same phosphorylation pattern could cause differential effects in the other tau isoforms, possibly through varied conformational effects. Using in vitro techniques, we examined the electrophoretic mobility, aggregation properties, and microtubule stabilization of all isoforms and their pseudophosphorylated counterparts. We found that pseudophosphorylation affected each isoform, but in several cases certain isoforms were affected more than others. These results suggest that hyperphosphorylation of tau isoforms could play a major role in determining the isoform composition of tau aggregates in disease.