DISPLAY OF PEPTIDES AND SINGLE-CHAIN ANTIBODIES ON BACTERIOPHAGE MS2 VIRUS-LIKE PARTICLES

Phage display is a powerful technology for selection of novel binding functions from large populations of peptide or antibody fragments. From a sufficiently complex library, phage bearing peptides or antibodies with practically any desired binding activity can be physically isolated by affinity selection, and, since each particle carries in its genome the genetic information for its own replication, the selectants can be amplified in bacteria. Existing display systems are based mostly on the filamentous phage M13. It is a powerful technology, but it has limitations for certain applications. Our work has concentrated on development of a new display platform based on virus-like particles (VLPs) of bacteriophage MS2, which addresses some of these limitations. The work presented here has two main foci - the display of single-chain antibody fragments (scFvs) on the MS2 VLP surface and the affinity selection of epitopes and epitope mimic from random sequence peptide libraries using monoclonal antibodies as targets. Here we demonstrate the display of several different scFvs via genetic fusion at the C-terminus of the MS2 coat protein single-chain dimer, including the following: M18 (scFv against anthrax protective antigen), AF-20 (scFv against AF-20 antigen, found on hepatocellular carcinoma cells), and scFv26/scFv66 (scFvs against Nipah virus G and F protein, respectively). Work with M18 demonstrates successful scFv incorporation into the MS2 VLP and intact antibody structure and function. VLPs displaying scFv AF-20 were used in both confocal microscopy and FACS experiments to demonstrate that these scFv-bearing VLPs can be specifically targeted to cells expressing the receptor. The functionality of VLPs displaying scFv26 or scFv66 was demonstrated in neutralization assays with NiV G protein-pseudotyped vesicular stomatitis virus (VSV). We also demonstrate the affinity selection of epitopes and epitope mimics from libraries of random sequence peptide displayed within the AB-loop of the MS2 VLP. Several different antibodies recognizing both protein and carbohydrate antigens were used as the selection targets. They included the following: the 2C7 and 2-1-L8 (monoclonal antibodies directed to lipooligosaccharide of Neisseria gonorrhoeae), MDVP-55A and GTX29202 (antibodies against a discontinuous epitope of the envelope protein of Dengue virus); MCA5792 (an antibody against the peptidoglycan of Staphylococcus aureus); and 2H1 (an antibody to the capsular glucuronoxylomannan (GXM) of the fungus Cryptococcus neoformans, and SYA/J6 (which recognizes the lipopolysaccharide of Shigella flexneri). From these selections, families of potential mimotopes were developed, and tested for activity with the selecting antibody and (in some cases) were used in mouse immunizations to attempt to promote an immune response against the original antigen. In the cases where immunizations were attempted, high-titer antibodies were generated that bound to the original antigen, suggesting the suitability of the MS2 VLP platform for identification of mimotopes for diverse epitope types

Evaluación de la Metodología 5S en la carpintería MORALES S.A.C. - Chimbote 2019

El presente trabajo de investigación, tuvo como objetivo principal evaluar la metodología de las 5S en la carpintería MORALES SAC, Nuevo Chimbote con la finalidad de demostrar el estado actual de la metodología 5S, así mismo dar a conocer el estado actual con respecto a cada criterio que maneja la mencionada metodología. Por ende, el diseño del estudio fue no experimental- transversal aplicando un Cheklist para la evaluación de la situación actual de la carpintería. La evaluación se realizó mediante observación directa, como herramienta de evaluación se utilizó un Checklist para cada criterio, luego se reflejó los datos obtenidos en un diagrama radial para indicar el porcentaje de cumplimiento y por último se evaluó el porcentaje de cumplimiento en la metodología 5S. Posterior al procesamiento de los datos adquiridos en el desarrollo de la investigación se concluyó que el estado actual de la carpintería es crítico puesto que se obtuvo como resultado un 19% de cumplimiento según la metodología 5S. En ese sentido, se recomienda implementar la metodología 5S para mejorar los criterios de clasificación, orden, limpieza, estandarización y disciplina

Mejora de procesos para incrementar la productividad en la operación pelado de mango, Santa Sofía del Sur S.A.C. - Casma 2020

La presente investigación tuvo por finalidad la aplicación de la mejora de procesos en la operación de pelado de mango para incrementar la productividad de la empresa Santa Sofía del Sur S.A.C. Se utilizó una investigación del tipo aplicada del diseño pre experimental con un pre prueba y post prueba, así mismo la muestra involucro a la productividad de mango congelado. Para el diagnostico se realizó un cursograma para conocer las operaciones del proceso, identificando un tiempo improductivo de 35.71%, por tal motivo la productividad inicial no fue la adecuada obteniendo 39.84%, por tal motivo se plantearon mejoras en base a la metodología PHVA, en la etapa planear se planifico mediante el cursograma del área de pelado, el estudio de tiempos y la técnica de interrogatorio determinando la necesidad de capacitar al personal, en el hacer se desarrolló mediante un diagrama bimanual, analizando el mejoramiento en ambas manos, mejorando el estudio de tiempo en 1:22:27, el verificar se redujo las actividades improductivas en un 4.7%, el actuar se planteó un manual de procedimientos. Concluyendo que la aplicación de la mejora de procesos aumento la productividad de materia prima en un 4.39%

Relación entre la intención de continuidad y variables experienciales en compradores chatbot-asistidos, de 18 a 45 años en Lima

El presente estudio de investigación tiene como principal objetivo conocer la relación que existe entre la variable intención de continuidad en el e-commerce asistido por chatbot y las variables experienciales; calidad de información, calidad de servicio, utilidad percibida y satisfacción, en compradores de dispositivos tecnológicos para educación remota de 18 a 45 años de Lima Metropolitana durante el año 2022. Por ello, se empleó un método de investigación no experimental correlacional con el fin de estudiar y contrastar las hipótesis planteadas, donde se evalúa el nivel de relación entre cada una de las variables en estudio. Asimismo, con la finalidad de recolectar datos se aplicaron cuestionarios virtuales, donde 331 personas participaron, pero solo 200 pasaron el filtro de elegibilidad. Cabe señalar que, el instrumento estuvo conformado por 21 ítems, es decir, 21 preguntas en escala de Likert, los cuales presentaron un nivel de confiabilidad de 0,936 en su totalidad. Finalmente, los principales hallazgos obtenidos permiten conocer más el comportamiento del consumidor en la experiencia de búsqueda de información hasta la post venta según el público objetivo del E-commerce, ya que se encontró patrones en cada uno de los segmentos generacionales.The main objective of this research study is to know the relationship that exists between the variable intention to continue in e-commerce assisted by chatbot and the experiential variables, quality of information, quality of service, perceived utility and satisfaction, in buyers of technological devices for remote education. from 18 to 45 years of Metropolitan Lima during the year 2022. Therefore, a correlational non-experimental research method was used to study and contrast the hypotheses, where the level of relationship between each of the variables in study. Likewise, to collect data, virtual surveys were applied, where 331 people participated, but only 200 passed the eligibility filter. It should be noted that the instrument consisted of 21 items, that is, 21 questions on the Likert scale, which presented a reliability level of 0.936 in its entirety. Finally, the main results obtained allow us to learn more about consumer behavior in the information search experience up to post-sale according to the target audience of E- commerce, since patterns were found in each of the generational segments.Tesi

Industrial antifoam agents impair ethanol fermentation and induce stress responses in yeast cells

Abstract The Brazilian sugarcane industry constitutes one of the biggest and most efficient ethanol production processes in the world. Brazilian ethanol production utilizes a unique process, which includes cell recycling, acid wash, and non-aseptic conditions. Process characteristics, such as extensive CO2 generation, poor quality of raw materials, and frequent contaminations, all lead to excessive foam formation during fermentations, which is treated with antifoam agents (AFA). In this study, we have investigated the impact of industrial AFA treatments on the physiology and transcriptome of the industrial ethanol strain Saccharomyces cerevisiae CAT-1. The investigated AFA included industrially used AFA acquired from Brazilian ethanol plants and commercially available AFA commonly used in the fermentation literature. In batch fermentations, it was shown that industrial AFA compromised growth rates and glucose uptake rates, while commercial AFA had no effect in concentrations relevant for defoaming purposes. Industrial AFA were further tested in laboratory scale simulations of the Brazilian ethanol production process and proved to decrease cell viability compared to the control, and the effects were intensified with increasing AFA concentrations and exposure time. Transcriptome analysis showed that AFA treatments induced additional stress responses in yeast cells compared to the control, shown by an up-regulation of stress-specific genes and a down-regulation of lipid biosynthesis, especially ergosterol. By documenting the detrimental effects associated with chemical AFA, we highlight the importance of developing innocuous systems for foam control in industrial fermentation processes.</jats:p

Effect of NRG1, GDNF, EGF and NGF in the Migration of a Schwann Cell Precursor Line

The Schwann cells are the myelinating glia of the peripheral nervous system that originated during development from the highly motile neural crest. However, we do not know what the guidance signals for the Schwann cell precursors are. Therefore, we set to test some of the known neurotrophins that are expressed early in developing embryos and have been shown to be critical for the survival and patterning of developing glia and neurons. The goal of this study was to determine more specifically if GDNF, NRG1 and NGF are chemoattractants and/or chemokinetic molecules for a Schwann cell precursor line, the Spl201. We performed live chemoattraction assays, with imaging and also presented these molecules as part of their growing substrate. Our results show for the first time that GDNF and NRG1 are potent chemoattractive and chemokinetic molecules for these cells while NGF is a chemokinetic molecule stimulating their motility