RESUMEN
In this study, nanoscale hydration dynamics of DNA-lipid blend dry films are investigated via small angle X-ray diffraction. Compared to the hydration of lipid films, fragmented short DNA strands and counterions in stacked lipid layers dramatically accelerate both the relaxation of the lamellar distance to a metastable interval and the subsequent peeling-off process of lipid bilayers. Moreover, genome-sized long DNA and counterions accelerate the relaxation process, but suppress the peeling-off process and simultaneously induce a damped-oscillation of the lamellar interval; this is probably due to the viscoelastic properties of the entangled long DNA dissolved in hydrated water between the stacked lipid bilayers. This study's findings can pave the way for producing cell-sized liposomes, which efficiently encapsulate any arbitrary sized DNA through natural swelling.
Asunto(s)
ADN/química , Membrana Dobles de Lípidos/química , Agua/química , Tamaño de la Partícula , Fosfatidilcolinas/química , Dispersión del Ángulo Pequeño , Difracción de Rayos XRESUMEN
What role does water play in the self-assembly of soft materials? To understand the correlation between the hydration state and the various self-assembled structures of a nonionic surfactant, terahertz time-domain spectroscopy has been performed for a C(12)E(5) solution with complementary use of small-angle x-ray scattering. Precise observations of the hydration state show clearly that transitions of the hydration state are accompanied by structural phase transitions of the surfactant from hexagonal to micelle to lamellae. These transitions of hydration state suggest that water is not a homogeneous solvent, and the interaction between water and the soft material is important for self-assembly.
Asunto(s)
Tensoactivos/química , Agua/química , Micelas , Dispersión del Ángulo Pequeño , Análisis Espectral , Electricidad Estática , Difracción de Rayos XRESUMEN
The hydration state of biomolecules is believed to affect their self-assembly. The hydration state of phospholipid bilayers is studied precisely by terahertz spectroscopy, by which water perturbed by a lipid membrane is detected sensitively from the observation of the relaxation dynamics of water molecules in the subpicosecond time scale. Combined with x-ray observation of the lamellar structure of the lipid, a long-range hydration effect on up to 4-5 layers of water is confirmed. Most water molecules in the lamellae fall into the hydration water, and condensation of them is also indicated.
Asunto(s)
Dimiristoilfosfatidilcolina/análogos & derivados , Membrana Dobles de Lípidos/química , Modelos Químicos , Fosfolípidos/química , Espectroscopía de Terahertz/métodos , Agua/química , Dimiristoilfosfatidilcolina/química , Dispersión del Ángulo Pequeño , Factores de Tiempo , Difracción de Rayos XRESUMEN
The mechanism for the formation of solid-supported phospholipid membranes during a drying process was investigated. Terracelike multilamellar structures were found to develop from a micellar solution with either spinodal decompositionlike process or nucleation growth, depending on the evaporation rate of an organic solvent. In contrast to the well-known kinetics of phase separation, fast drying induces nucleation while slow drying induces spinodal decompositionlike lipid-film formation. The existing models for the interpretation of phase separation are not sufficient to understand this unexpected kinetics. We suggest a schematic model with which this kinetic feature can be interpreted in terms of a self-assembly pathway in a three-component phase diagram for a phospholipid, organic solvent, and water.
Asunto(s)
Desecación/métodos , Membrana Dobles de Lípidos/química , Fluidez de la Membrana , Membranas Artificiales , Modelos Químicos , Fosfolípidos/química , Simulación por Computador , Cinética , Transición de FaseRESUMEN
Membrane fluctuations of dipalmitoylphosphatidylcholine (DPPC) and dimyristoylphosphatidylcholine (DMPC) were investigated by neutron spin echo spectroscopy. The intermediate structure factor was analyzed in terms of the model proposed by Zilman and Granek (Phys. Rev. Lett. 77, 4788 (1996)), and the bending modulus of lipid bilayers was derived. The hardening of a lipid bilayer upon approaching the main transition point in the anomalous swelling regime was observed, which naturally connects the bending modulus in the gel phase below the main transition temperature.
Asunto(s)
Membrana Dobles de Lípidos/química , Neutrones , 1,2-Dipalmitoilfosfatidilcolina/química , Bromuros/química , Cloruro de Calcio/química , Cristalización , Óxido de Deuterio/química , Dimiristoilfosfatidilcolina/química , Compuestos de Potasio/químicaRESUMEN
Thirty-five monoclonal antibodies (MAbs) against glycoprotein (G protein) of the RC-HL strain of the rabies virus have been established. Using these MAbs, two antigenic sites (I and II) were delineated on the G protein of the RC-HL strain in a competitive binding assay. Of these, 34 MAbs recognized the epitopes on site II. Site II was further categorized into 10 subsites according to their patterns in a competitive binding assay. Each site II-specific MAb showed 5 to 23 nonreciprocal competitions. The reactivities of 35 MAbs to rabies and rabies-related viruses in an indirect immunofluorescent antibody test showed that six MAbs in group A binded to rabies and rabies-related viruses and eight MAbs in group E reacted only with rabies viruses, considering that the former represent the genus-specific of Lyssavirus and the latter are rabies virus-specific. From biological assays, 28 of the 35 MAbs showed neutralization activity, 31 showed hemagglutination inhibition (HI) activity, and 18 showed immunolysis (IL) activity. The MAbs recognizing neutralization epitopes fell into at least three groups: those exhibiting both HI and IL activity, those showing only HI activity, and those showing neither HI nor IL activity. All IL epitopes overlap with HA epitopes. Five of the nine MAbs which reacted with the antigen treated by sodium dodecyl sulfate in ELISA were not reduced, or reduced only slightly, in the titer. None of the MAbs reacted with 2-mercaptoethanol-treated antigen. Only one MAb that recognized site I reacted with the denatured G protein in a Western blotting assay, indicating that its epitope is linear. These results suggest that almost all of the epitopes on the G protein of the rabies virus are conformation-dependent and the G protein forms a complicated antigenic structure.
Asunto(s)
Antígenos Virales/inmunología , Glicoproteínas/inmunología , Virus de la Rabia/inmunología , Proteínas del Envoltorio Viral/inmunología , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Unión Competitiva , Ensayo de Inmunoadsorción Enzimática , Mapeo Epitopo , Técnica del Anticuerpo Fluorescente Indirecta , Pruebas de Inhibición de Hemaglutinación , Pruebas de Neutralización , Desnaturalización ProteicaRESUMEN
A set of 29 monoclonal antibodies (MAbs) specific for the rabies virus nucleoprotein (N protein) was prepared and used to analyze the topography of antigenic sites. At least four partially overlapping antigenic sites were delineated on the N protein of rabies virus by competitive binding assays. Indirect immunofluorescent antibody tests using MAbs with a series of rabies and rabies-related viruses showed that epitopes shared by various fixed and street strains of rabies virus were mainly localized at antigenic sites II and III, while epitopes representing the genus-specific antigen of Lyssavirus were widely presented at sites I, III and IV. All but one of seven MAbs specific for antigenic sites I, IV and bridge site (I and II) reacted with the antigen that had been denatured by sodium dodecyl sulfate or 2-mercaptoethanol, as well as with the denatured N protein in Western blotting assays. However, none of the MAbs against antigenic sites II and III reacted with the denatured antigen. These data indicate that antigenic sites I and IV, and sites II and III on the N protein of rabies virus are composed of linear and conformation-dependent epitopes, respectively.
Asunto(s)
Cápside/inmunología , Epítopos/inmunología , Virus de la Rabia/inmunología , Proteínas del Núcleo Viral/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Unión Competitiva/inmunología , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Mercaptoetanol/farmacología , Ratones , Ratones Endogámicos BALB C , Conformación Proteica , Desnaturalización Proteica/efectos de los fármacos , Ensayo de Radioinmunoprecipitación , Dodecil Sulfato de Sodio/farmacologíaRESUMEN
The effects of beryllium addition to a dental cobalt-chromium alloy on biological compatibility as well as physical properties were examined and the following results were obtained. 1. Slight, but significant depression of the rates of cell multiplication was obtained with the experimental groups, i.e., alloy with no beryllium added as well as alloys with beryllium of up to 3.0 per cent by weight, compared to control group, which contained no alloys, but a glass disk. Within the experimental groups no significant difference in the rates of cell multiplication was found between the alloys with beryllium addition ranging from zero up to 2.0% by weight. However, alloy with 3.0% beryllium yielded slight, but significant depression of the rates of cell multiplication. Pure beryllium metal revealed severe cytotoxicity. 2. Cell morphology of the experimental groups confirmed the above results of the rate of cell multiplication. 3. Increase of beryllium within the alloys resulted in increase of tensile strength as well as Rockwell hardness, while elongation and fusion temperature were brought down. 4. Metallographs of alloys and cast specimens confirmed the results of the mechanical properties. The more beryllium was added, the smaller was the alloy crystal observed. 5. Loss of beryllium in the alloy was found during such procedures as melting each metal for making up alloys and casting. It is considered that the present results will be able to lend suggestions to beryllium use in dentistry with regard to biological compatibility as well as physical properties.