RESUMEN
PIEZO1 is a mechanosensitive channel that converts applied force into electrical signals. Partial molecular structures show that PIEZO1 is a bowl-shaped trimer with extended arms. Here we use cryo-electron microscopy to show that PIEZO1 adopts different degrees of curvature in lipid vesicles of different sizes. We also use high-speed atomic force microscopy to analyse the deformability of PIEZO1 under force in membranes on a mica surface, and show that PIEZO1 can be flattened reversibly into the membrane plane. By approximating the absolute force applied, we estimate a range of values for the mechanical spring constant of PIEZO1. Both methods of microscopy demonstrate that PIEZO1 can deform its shape towards a planar structure. This deformation could explain how lateral membrane tension can be converted into a conformation-dependent change in free energy to gate the PIEZO1 channel in response to mechanical perturbations.
Asunto(s)
Microscopía por Crioelectrón , Canales Iónicos/química , Canales Iónicos/ultraestructura , Microscopía de Fuerza Atómica , Silicatos de Aluminio/química , Animales , Células HEK293 , Humanos , Canales Iónicos/metabolismo , Liposomas/química , Liposomas/metabolismo , Liposomas/ultraestructura , RatonesRESUMEN
Using membrane processes we studied the improvement of the treatment of waste emulsified by mineral oils and alkylphenolethoxylate (APE) and investigated diatomaceous earth processing for comparison. A quantitative analysis of the mineral oils and APE using column chromatography was applied to assess these processes. Despite substantial rejection of the turbidity values of emulsified model waste, the percentages of mineral oil and APE rejections using microfiltration (MF) processes were 39-61% and 16-19%, respectively. In contrast, the ultrafiltration (UF) process could reject 97% for the mineral oils and 90% for APE. The diatomaceous earth adsorption processes reduced the mineral oils 40-49% and the APE, 8-14%, and therefore were inferior to the UF process for waste treatment. The average fluxes of both the MF and UF membranes until the volume reduction factor (VRF) reached a value of 5 were almost the same in the range of 8 x 10(-6)-14 x 10(-6) m3 m-2 s-1. For commercial application, decrease in the permeate flux by fouling needs to be resolved.
Asunto(s)
Aceite Mineral/química , Tensoactivos/química , Eliminación de Residuos Líquidos/métodos , Purificación del Agua/métodos , Adsorción , Tierra de Diatomeas/química , Filtración , Membranas Artificiales , Fenoles/químicaRESUMEN
This article presents the case of a Japanese woman who had Ekman-Westborg-Julin trait. She had general macrodontia with multituberculism, evagination of the premolar, single conical roots, shovel-shaped incisors, enamel hypoplasia, impacted tooth, dental crowding, and an open bite. The oral and general characteristics of this patient are described and include the histological and radiographic findings of the mandibular third molars. We suggest that the distinctive oral features with macrodontia of the permanent teeth, multituberculism, evagination, single conical roots, and impaction of the tooth could be defined as the Ekman-Westborg-Julin trait.
Asunto(s)
Anomalías Dentarias/patología , Adolescente , Diente Premolar/anomalías , Diente Canino/anomalías , Dens in Dente/patología , Hipoplasia del Esmalte Dental/patología , Femenino , Estudios de Seguimiento , Humanos , Incisivo/anomalías , Maloclusión/patología , Diente Molar/anomalías , Mordida Abierta/patología , Síndrome , Corona del Diente/anomalías , Raíz del Diente/anomalías , Diente Impactado/patologíaRESUMEN
Oral manifestations of Down syndrome include high susceptibility to gingival inflammation with early onset and rapidly progressive periodontitis. The influence of reactive oxygen species (ROS) on periodontitis of Down syndrome is unclear. The aim of this study was to characterize ROS formation in Down syndrome-gingival fibroblasts (DS-GF) using electron spin resonance (ESR) spin trapping with 5,5-dimetyl-1-pyrolline-N-oxide (DMPO), and to determine whether ROS generation plays a role in the pathogenesis of periodontitis in Down syndrome patients. We observed formation of the DMPO-OH spin adduct, indicating HO* generation from cultured DS-GF and non-DS-GF. The increased HO* generation in cultured DS-GF was strongly decreased in the presence of the H2O2 scavenger, catalase, or the iron chelator, desferal. This may due to the enzymatic ability of over-expressed CuZn-superoxide dismutase in Down syndrome to catalyze the formation of H2O2 from O2*-, thereby increasing the availability of substrate H2O2 for the iron-dependent generation of HO* via the Fenton reaction, suggesting that HO* generated from DS-GF may be involved in progressive periodontitis of Down syndrome.
Asunto(s)
Síndrome de Down/metabolismo , Espectroscopía de Resonancia por Spin del Electrón/métodos , Fibroblastos/metabolismo , Encía/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Adulto , Catalasa/metabolismo , Catalasa/farmacología , Técnicas de Cultivo de Célula , Deferoxamina/farmacología , Síndrome de Down/patología , Femenino , Fibroblastos/citología , Encía/citología , Humanos , Peróxido de Hidrógeno/metabolismo , Radical Hidroxilo/metabolismo , Masculino , Modelos Químicos , Oxidación-Reducción/efectos de los fármacos , Estrés Oxidativo , Periodontitis/metabolismo , Periodontitis/patología , Superóxido Dismutasa/metabolismo , Superóxidos/metabolismoRESUMEN
Actinobacillus actinomycetemcomitans, a Gram-negative periodontopathic bacterium, produces a leukotoxin belonging to the RTX family. The production of leukotoxin varies greatly among different strains of this species. In this paper the effects of growth rate and bicarbonate on the leukotoxin production by a toxin-production-variable strain (301-b) during growth in a chemostat were examined. When the bacterium was grown in anaerobic fructose-limited chemostat cultures (pH 7.0 and 37 degrees C) at dilution rates (D) ranging from 0.04 to 0.20 h-1 in the absence and presence of 10 mM bicarbonate, it produced leukotoxin as a cluster of two polypeptides (M(r) 113,000 and 120,000) and complexed with nucleic acids on the bacterial cell surface. The relationship between leukotoxin production and specific growth rate was analysed by plotting the specific rate of leukotoxin production [qLT, in microgram (mg dry wt)-1 h-1] against D. The plots were approximated to the linear relationships qLT = 2.7D-0.058 and qLT = 9.3D-0.407 without and with bicarbonate, respectively. These relationships suggest that the apparent leukotoxin production is a result of both growth-rate-dependent production and growth-rate-independent decomposition. The cellular leukotoxin level was also followed after the change from chemostat to batch culture in the same fermenter. In batch culture leukotoxin production stopped immediately and the cellular toxin level rapidly decreased, suggesting toxin decomposition. From the slopes of the approximated linear relationships between qLT and D, a theoretical maximum leukotoxin yield (YLT) was estimated as 2.7 and 9.3 micrograms (mg dry wt)-1 in the absence and presence of 10 mM bicarbonate, respectively. The increased YLT value in the cultures containing bicarbonate indicated that the addition stimulated the efficiency of leukotoxin synthesis up to about threefold. Further increases of bicarbonate concentration to between 20 and 40 mM had no effect on the total leukotoxin production, but the amount of extracellular leukotoxin increased with higher bicarbonate concentrations.