RESUMEN
Glycinin is an important allergenic protein. A1a is the acidic chain of the G1 subunit in glycinin (G1A1a), and it has strong allergenicity. In this study, we used phage display technology to express the protein of G1A1a and its overlapping fragments and an indirect enzyme-linked immunosorbent assay (iELISA) to determine the antigenicity and allergenicity of the expressed protein. After three rounds of screening, it was determined that fragment A1a-2-B-I (151SLENQLDQMPRRFYLAGNQEQEFLKYQQEQG181) is the allergenic domain of G1A1a destroyed by thermal processing. In addition, three overlapping peptides were synthesized from fragments A1a-2-B-I, and a linear epitope was found in this domain through methods including dot blot and iELISA. Peptide 2 (157DQMPRRFYLANGNQE170) showed allergenicity, and after replacing it with alanine, it was found that amino acids D157, Q158, M159, and Y164 were the key amino acids that affected its antigenicity, while Q158, M159, R162, and N168 affected allergenicity.
Asunto(s)
Alérgenos , Globulinas , Calor , Proteínas de Soja , Alérgenos/inmunología , Alérgenos/química , Humanos , Globulinas/química , Globulinas/inmunología , Proteínas de Soja/química , Proteínas de Soja/inmunología , Secuencia de Aminoácidos , Hipersensibilidad a los Alimentos/inmunología , Epítopos/química , Epítopos/inmunología , Dominios Proteicos , Antígenos de Plantas/inmunología , Antígenos de Plantas/química , Antígenos de Plantas/genética , Glycine max/química , Glycine max/inmunología , Ensayo de Inmunoadsorción EnzimáticaRESUMEN
Glycinin is an important allergen in soybeans. In this study, molecular cloning and recombinant phage construction were performed to explore the antigenic sites of the glycinin A3 subunit that were denatured during processing. Next, the A-1-a fragment was located as the denatured antigenic sites by indirect ELISA. The combined UHP heat treatment showed better denaturation of this subunit than the single heat treatment assay. In addition, identification of the synthetic peptide showed that the A-1-a fragment was an amino acid sequence containing a conformational and linear IgE site, in which the first synthetic peptide (P1) being both an antigenic and allergenic site. The results of alanine-scanning showed that the key amino acids affecting antigenicity and allergenicity of A3 subunit were S28, K29, E32, L35 and N13. Our results could provide the basis for further development of more efficient methods to reduce the allergenicity of soybeans.
Asunto(s)
Globulinas , Proteínas de Soja , Proteínas de Soja/química , Glycine max , Globulinas/química , Secuencia de Aminoácidos , AlérgenosRESUMEN
Soft exo-suit could facilitate walking assistance activities (such as level walking, upslope, and downslope) for unimpaired individuals. In this article, a novel human-in-the-loop adaptive control scheme is presented for a soft exo-suit, which provides ankle plantarflexion assistance with unknown human-exosuit dynamic model parameters. First, the human-exosuit coupled dynamic model is formulated to express the mathematical relationship between the exo-suit actuation system and the human ankle joint. Then, a gait detection approach, including plantarflexion assistance timing and planning, is proposed. Inspired by the control strategy that is used by the human central nervous system (CNS) to handle interaction tasks, a human-in-the-loop adaptive controller is proposed to adapt the unknown exo-suit actuator dynamics and human ankle impedance. The proposed controller can emulate human CNS behaviors which adapt feedforward force and environment impedance in interaction tasks. The resulting adaptation of actuator dynamics and ankle impedance is demonstrated with five unimpaired subjects and implemented on a developed soft exo-suit. The human-like adaptivity is performed by the exo-suit in several human walking speeds and illustrates the promising potential of the novel controller.
Asunto(s)
Dispositivo Exoesqueleto , Robótica , Humanos , Tobillo/fisiología , Articulación del Tobillo/fisiología , Impedancia Eléctrica , Caminata/fisiología , Fenómenos BiomecánicosRESUMEN
Glycinin is an important storage protein in soybean, but it can also lead to allergic reactions in humans. In this study, based on a low-cost, simple, rapid and portable lateral flow immunoassay test strip, combined with high sensitivity surface enhanced Raman spectroscopy (SERS) technology, a sandwich lateral flow immunochromatographic test strip for rapid detection of soybean allergen glycinin is established. In the experiment, colloidal gold was covalently conjugated with rabbit-derived polyclonal antibodies of glycinin and Raman probe molecule 4-aminothiophenol(4-PATP) to prepare the immunoprobe. The respective optimal PATP and optimal antibody labeling amounts of colloidal gold solution were 1.05 × 10-2mol/L and 4.6 × 10-8mol/L. The detection limit of the test strip for glycinin was 4.87 ng/mL. The recovery rate ranged from 91 to 107 % and the CV was between 3 and 10 %. The test strip underwent no cross-reaction with ß-conglycinin, sesame protein, peanut protein, wheat protein or whey protein. The results of the experiment showed that this method exhibits high sensitivity and specificity.
Asunto(s)
Oro Coloide , Espectrometría Raman , Humanos , Animales , Conejos , Oro Coloide/química , Espectrometría Raman/métodos , Anticuerpos Monoclonales/química , Inmunoensayo/métodos , Glycine maxRESUMEN
Wearable walking exoskeletons show great potentials in helping patients with neuro musculoskeletal stroke. Key to the successful applications is the design of effective walking trajectories that enable smooth walking for exoskeletons. This work proposes a walking planning method based on the divergent component of motion to obtain a stable joint angle trajectory. Since periodic and nonperiodic disturbances are ubiquitous in the repeating walking motion of an exoskeleton system, a major challenge in the walking control of wearable exoskeleton is the joint angle drift problem, that is, the joint angle motion trajectories are not necessarily periodic due to the presence of disturbance. To address this challenge, this work develops an adaptive repetitive control strategy to guarantee that the motion trajectories of joint angle are repetitive. In particular, by treating the disturbance as system uncertainties, an adaptive controller is designed to compensate for the uncertainties based on an integral-type Lyapunov function. A fully saturated learning approach is then developed to achieve asymptotic tracking of repetitive walking trajectories. Extensive experiments are carried out to demonstrate the effectiveness of the tracking performance.
RESUMEN
This article presents a human-in-the-loop cooperative control of a walking exoskeleton to provide assistance to the user and enhance human mobility. First, a dynamic mathematical model of the human-exoskeleton system is derived, and then, a human-in-the-loop cooperative control framework is proposed in two ways: separable cooperative control (SCC) and interactive cooperative control (ICC), respectively. The SCC introduces a space division of the human and the exoskeleton, while the ICC allows the robot to perceive the human intention and follows human motor, thereby improving the collaborative performance. The ICC formulates the impedance connection between the human and the exoskeleton in the divided orthogonal subspaces of walking, such that the robot is able to modify its position of center of mass (COM) when its motor trajectory deviates the one of the human. In addition, a novel adaptation control is proposed to deal with the unmodeled dynamics and trajectory tracking. Finally, to validate the effectiveness of our proposed controller, a series of experiments are conducted in three adults in gait at different speeds. It shows that the proposed controller can preserve the periodic walking gait and inherit the robustness of dealing with perturbations during walking.