Ciencias Exactas y Ciencias de la Salud
Permanent URI for this collectionhttps://hdl.handle.net/11285/551039
Pertenecen a esta colección Tesis y Trabajos de grado de las Maestrías correspondientes a las Escuelas de Ingeniería y Ciencias así como a Medicina y Ciencias de la Salud.
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- Immunoaffinity aqueous two-phase systems to establish novel bioprocesses for the primary recovery of CD133+ stem cells(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2018-05-25) Ornelas González, Alonso; Rito Palomares, Marco Antonio; Zavala Arcos, Judith; González González, Mirna Alejandra; Rito Palomares, Marco Antonio; Zavala Arcos, Judith; González González, Mirna AlejandraA short processing time and efficient scale-up stem cell isolation bioprocess is essential to exploit the potential of these cells for the treatment of multiple chronic diseases. Various methodologies have been used for stem cell recovery, however, most of them present economical and/or time-consuming drawbacks. In this work, the characterization and optimization of immunoaffinity aqueous two-phase systems, a liquid-liquid based separation technology enhanced with the PEGylation of the antibody, was conducted with the aim of increasing the specificity for the recovery of CD133+ stem cells from human umbilical cord blood samples. The methodology consisted in evaluating the partitioning of the different PEGylated antibodies (amine, carboxyl, thiol, succinimidyl ester, methoxy PEG and maleimide) in three previously studied aqueous two-phase systems (ATPS); PEG-dextran (DEX), Ucon-DEX and Ficoll-DEX. Subsequently, an optimization step was accomplished to manipulate the partition behavior of the CD133/2-pure antibody to the desired phase in the selected systems by varying (increasing and decreasing) two parameters closely related with the partitioning of molecules in aqueous two-phase systems; tie-line length (TLL) and volume ratio (VR). Afterwards, the partitioning behavior of the six different PEGylated antibodies in the optimized systems was tested. According to the results, the PEGylation of the CD133/2-biotin antibody induced a favorable change with respect to the non-PEGylated one when Ucon-DEX system was used, fractionating it to both phases. Likewise, the optimization of the systems showed to be effective to induce a change in the partition preference of the antibody. The best results were obtained when Ucon-DEX or PEG-DEX systems with TLL 15% w/w or 20% w/w with VR 3 were combined. Finally, PEGylated antibodies were added to the selected optimized systems. Even though a shift in the fractionation preference of the PEGylated CD133/2-biotin antibody was achieved in the optimized systems, it was not the adequate partition to justify the evaluation of this immunoaffinity ATPS with human umbilical cord samples. Both PEGylation and optimization showed to be effective to induce a change in the partition preference of the antibody, however, further studies are required to find the optimal system composition that will fractionate 100% of the antibody to the contaminants opposite phase, making this system an ideal candidate to be tested for the selectivity of CD133+ stem cells.
- Effect of ultrasound on protein yield and fate of alkaloids during lupin alkaline extraction process(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2018-05-14) Aguilar Acosta, Luis Alberto; Chuck Hernández, Cristina Elizabeth; Othón Serna Saldívar, Sergio Román; Benavidez Lozano, Jorge Alejandro; Rodríguez Rodríguez, JoséProtein isolates from legumes have attracted scientific attention because they are an efficient option to cover the daily protein requirement. One of the legumes with the highest protein content (20-50%) nowadays is Lupinus spp., with a good amino acid profile and moderate protein digestibility (≈ 70%) but with the presence, in certain species, of quinolizidine alkaloids (QA), toxic to the organism. The most popular method to QA removal is one based in washing with water, but inconvenient because of the high use of water and time. With the aim to propose eventually a new process for lupin detoxification, the objective of this work was the elucidation of the fate of QA during lupin protein extraction process assisted with ultrasound and the evaluation of the nutritional and functional properties of the protein fraction. Proximal characterization, concentration of anti-nutritional compounds, amino acid profile and protein solubility profile of flours from three lupin species were assessed: L. albus, L. angustifolious and L. mutabilis. The result showed a significant difference (p < 0.05) in protein concentration, total alkaloids and particle size between the three species flours. Considering the above, the species L. mutabilis and L. angustifolious were chosen to study the behavior of the protein fraction in terms of functionality (water absorption and nitrogen solubility), composition (protein yield and percentage reduction of alkaloids during the extraction process) and resistance to thermal treatments (displacement of denaturation temperature, change in secondary structure and modification of the electrophoretic profile). The results obtained for L. mutabilis described the ultrasound effect as beneficial for protein yield (14% more than control with 10 min ultrasound) and QA reduction from bagasse (81% less than control with 10 min ultrasound treatment) and protein isolate (50% less than control with 10 min ultrasound treatment). In addition, the change in protein structure and composition modified the functional and thermal properties of the protein, making the isolate a good candidate for food ingredient. In the other hand, L. angustifolious was more resistant to the ultrasound effect with no significant difference between treatments (10 and 15 min) and control but with the lower toxicity (3 ppm of QA in control protein isolate) and better amino acid score (0.85 of 15 min ultrasound treatment). These results will be useful to design processes to assist in the objective for meet protein demand of the population.
- Effects of uvb light, wounding stress and storage time on the biosynthesis of betalains, phenolic compounds and ascorbic acid in red prickly pears (opuntia ficus-indica cv. Rojo vigor)(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2018-05-14) Ortega-Hernández, Erika; Jacobo Velázquez, Daniel Alberto; Benavides-Lozano, Jorge; Cisneros-Zevallos, Luis; Welti-Chanes, JorgeUVB light In the past years, abiotic stresses such as wounding and ultraviolet radiation have been studied as an effective tool to improve the accumulation of bioactive phytochemicals in fresh produce. Postharvest application of UVB-radiation and wounding stress induces the accumulation of secondary metabolites in horticultural crops. However, there are few studies evaluating the combined effects of both stresses Therefore, the present thesis evaluated, on the one hand, the separated and combined effects of UVB radiations and wounding stress on the total content of betalains, phenolic compounds and ascorbic acid in red prickly pear (Opuntia Ficus-Indica cv. Rojo Vigor) treated with UVB light (6.4 W•m-2) for 0, 15, 90 and 180 min, before and after storage (24 h at 16 °C); and on the other hand, the effects of UVB radiations and wounding stress, alone or in combination, on the profile of individual betalains and phenolic compounds, as well as the activity of the enzymes involved in their biosynthesis, in red prickly pear treated with UVB light (6.4 W•m-2) for 0 and 15 min, before and after storage (24 h at 16 °C). The highest phenolic accumulation occurred 24 h after UVB treatment (15 min) in whole pulp and peel, increasing by 125.8% and 33.8%, respectively. In the whole fruit pulp, there was a significant increment in kaempferol (64.5%), sinapoyl malate (187.7%), 1-O-β-D-Glucopyranosyl sinapate (57.7%), rosmarinic acid (584.5%), vanillic acid (15.1%), sinapic acid (23.3%), quercetin3-O-glucoside (28.4%) and kaempferol 3-O-sophoroside-7O-glucoside (57.7%). On the other hand, the whole fruit peel showed an increment in the content of kaempferol (631.5%), sinapoyl malate (172.7%), 1-O-β-D-Glucopyranosyl sinapate (505.7%), rosmarinic acid (394.5%), p-coumaric (53.9%), sinapic acid (639.3%), quercetin3-O-glucoside (133.4%), kaempferol 3-Oglucoside (160.9%), quercetin (425.2%) and kaempferol 3-O-sophoroside-7O-glucoside (63.7%). The increment in the PAL activity (1173.5% in pulp and 239.3% in peel) after 12 h of storage corresponds to the increment in content of phenolics. The highest betalain accumulation occurred 24 h after UVB treatment (15 min) in whole pulp and wounded peel, increasing by 49.8% and 33.1%, respectively. The whole fruit pulp showed a significant increment in muscaaurin VII (293.6%), indicaxanthin (316%), betanin (26.2%) and iso-betanin (70.6%); whereas vulgaxanthin III (-21.1%), betanidin (-78.9%), neobetanidin (-87.3%) and gomphrenin (-55.4%) decreased. On the other hand, the wounded fruit peel showed a significant increment in muscaaurin VII (33.2%), indicaxanthin (159.1%), betanin (161.8) and iso-betanin (98.7%). The increment in the TYR hydroxylase (744.1% in pulp and 332.9% in peel) and TYR oxidase activity (81.6% in pulp and 59.2% in peel), as well as cyclo- Dopa formation (637.2% in pulp and 439.3% in peel) after 12 h of storage corresponds to the increment in content of betalains. The highest ascorbic acid accumulation occurred 24 h after UVB treatment in wounded pulp and peel, increasing by 67.2% and 84.6%, respectively. The increment in the GalLDH activity (85.4% in pulp and 502.3% in peel) after 12 h of storage corresponds to the increment in content of ascorbic acid.Results indicated that the application UVB light alone or combined with wounding stress in red prickly pear can be used as an effective strategy to induce the accumulation of secondary metabolites with potential application in the food, dietary supplements and pharmaceutical industries; since these postharvest treatments are low cost, have no legal restrictions, and do not require complex equipment.
- Efecto de agentes reductores y/o procesamiento térmico sobre la actividad de inhibido de tripsina y solubilidad de concentrados proteicos de soya o garbanzo(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2017-12-07) Aviles Gaxiola, Sara; 1; 1; 1; Cristina Chuck Hernández; Sergio Román Othón Serna Saldívar; Jorge Alejandro Benavides Lozano; Escurla de Ingeniería y Ciencias; Campus MonterreySe evaluó el efecto de dos agentes reductores (metabisulfito de sodio o Lhcisteína) y/o tratamiento térmico (80 o 121°C) en la Actividad de Inhibidor de Tripsina (AIT) de soya y garbanzo. Las materias primas se caracterizaron desde el punto de vista fisicoquímico y además se determinó la actividad y peso molecular de los inhibidores de tripsina (ITs). En el análisis proximal se determinó que el mayor porcentaje en peso de la soya fue proteína, mientras que en garbanzo fueron los carbohidratos. Se identificó en soya un IT característico (≈ 20 kDa), mientras que en garbanzo se determinaron dos bandas electroforéticas con actividad de inhibidor no reportadas antes en la literatura (≈ 25 y 37 kDa). Al ejecutar los tratamientos de agentes reductores y temperatura, el más efectivo en soya fue la combinación de metabisulfito de sodio + 80ºC, mientras que en garbanzo fue Lhcisteína sin tratamiento térmico. Se lograron inactivaciones de 99.42 y 74.53 % respectivamente. Posterior a los tratamientos se evaluó la funcionalidad proteica de ambas materias primas. Se encontró que en soya, la solubilidad de agua se redujo en 44.24% mientras que la absorción de agua aumentó casi 4 veces respecto al control. Por otro lado, las propiedades de solubilidad e interacción con agua en garbanzo no se vieron afectadas de manera significativa. Respecto a la digestibilidad in vitro, se observó un ligero aumento en ambos materiales, sin embargo, no fue estadísticamente significativo (~1%). Tras los tratamientos de inactivación, se determinó que en garbanzo la estructura proteica secundaria no se modificó, mientras que en soya aumentó el porcentaje de ßhplegada, bucles, αhhélice y cadenas no ordenadas mientras que las estructuras agregadas se redujeron en 9%. Se obtuvo también el perfil electroforético de los concentrados proteicos tratados para inactivar IT y de la mano de los resultados anteriores, se observó que garbanzo presentó el mismo patrón de bandas con respecto a su control, mientras que la soya mostró una disminución de especies de bajo peso molecular y un incremento de las bandas por encima de 150 KDa. Al analizar los cambios a una mayor escala, la microestructura de los concentrados de soya o garbanzo no se vio afectada, mientras que las propiedades de flujo y la actividad de agua (aw) sí cambiaron de forma significativa. En conclusión, el mejor tratamiento para inactivar IT de leguminosas dependerá de la estructura propia de este factor anti nutrimental. En el caso de garbanzo el uso de Lhcisteína fue el más eficiente reduciendo la AIT, lo cual fue adecuado para mantener intactas las propiedades funcionales, la estructura secundaria y el perfil electroforético de las proteínas. En soya, el tratamiento térmico (80°C) combinado con metabisulfito de sodio tuvo el mayor efecto de inactivación de IT, sin embargo, esto cambió la estructura de las proteínas, su funcionalidad y en menor medida su perfil electroforético.
- Combined effect of ultrasound treatment and phytohormones on the accumulation of bioactive compunds in broccoli florets(2017-05-27) Aguilar Camacho, Miguel; Jacobo Velázquez, Daniel Alberto; Wrlti Chanes, Jorge Santos; Benavidez Lozano, Jorge AlejandroIn the past years, postharvest abiotic stresses such as wounding, modified atmospheres, UV-radiation, and the exogenous application of ethylene (ET) and methyl jasmonate (MJ) have been studied as an effective tool to improve the accumulation of bioactive compounds in fresh produce. In this context, previous reports in literature suggest that non-thermal processing technologies such as ultrasound (US) activate the biosynthesis of nutraceuticals in crops by a similar mechanism exerted by wounding stress. However, there is still limited information in literature regarding emerging technologies and their effect on the biosynthesis of bioactive compounds. Therefore, the present thesis evaluated the immediate and late response to US treatment (20 min, frequency 24 kHz, amplitude 100 μm) on glucosinolates, isothiocyanates, phenolic compounds and ascorbic acid in broccoli florets. Moreover, the combined effects of US with exogenous MJ (250 ppm) and/or ethylene ET (1000 ppm) on the content of bioactive compounds were also evaluated during storage (72 h at 15 ºC) of broccoli florets. As an immediate response to US treatment, the extractability of glucosinolates such as glucoraphanin (795%), 4-hydroxy glucobrassicin (153%), and glucobrassicin (78.6%) increased as compared with the control (CT), and induced the detection of gluconasturtiin (0.30 mmol/kg) not identified in the control. Furthermore, US treatment increased the extractability of 1- sinapoyl-2-feruloylgentiobiose (57.23%) as compared with the control, whereas the concentration of the other phenolic compounds remained unaltered. Similarly, the content of total ascorbic acid remained unchanged immediately after US treatment. The enhanced extractability of some bioactive compounds was attributed to cell disruption by US treatment. During storage, gluconasturtiin and 4-methoxy glucobrassicin, which were not detected in CT samples before storage, showed accumulation at 24 h (0.69 mmol/kg) and 72 h (0.29 mmol/kg) of storage, respectively. On the other hand, the content of glucoraphanin and 4- hidroxy glucobrassicin, which were immediately increased by US treatment, remained stable during storage in US samples. However, the high increased content of gluconasturtiin and 4- methoxy glucobrassicin at 24 h and 72 h of storage in CT samples, was not detected in US samples. The combined application of MJ and US in broccoli florets, induced a synergistic effect on the accumulation of 4-hydroxy glucobrassicin, glucoerucin, gluconasturtiin and neoglucobrassicin at 72 h of storage, showing 187.1%, 111.92%, 755.9% and 232.8%, respectively, higher content as compared with US 72 h samples. When ET was applied on US samples, it impeded the accumulation of gluconasturtiin at 24 h of storage, whereas the accumulation of 4-methoxy glucobrassicin (0.52 mmol/kg) was slightly induced at 72 h. Interestingly, when the three stressors where applied together (US+MJ+ET) the synergistic effect of US and MJ observed on the accumulation of 4-hydroxy glucobrassicin, glucoerucin, gluconasturtiin and neoglucobrassicin at 72 h of storage was repressed. Regarding with phenolic compounds the application of US increased the accumulation of 5-O-CQA (70.6%) and 1,2,2-TSG (56.4%) at 72 h and 24 h of storage, respectively, whereas the content of CA (29.8%) decreased at 48 h, as compared with CT samples. Similar to glucosinolates, after 72 h of storage, US and MJ showed a synergistic effect on the accumulation of 3-O-CQA, 1-S-2-FG, and 1,2,2-TSG, showing 73.4%, 56.0% and 136.7%, respectively, as compared with 72h CT+MJ samples. Interestingly, the application of ET did not modify the effect exerted by US on the accumulation of individual phenolic compounds. Likewise, the application of the three stressors in broccoli florets (US+MJ+ET samples) inhibited the accumulation of 3-O-CQA, 1-S-2-FG, and 1,2,2-TSG observed in US+MJ samples stored for 72 h. In general, the ascorbic acid content decreased in most samples during storage. Samples treated with US alone did not show significant differences in the ascorbic acid content during storage when compared with CT samples. However, when MJ+ET were applied, the content of total ascorbic acid was significantly reduced in CT+MJ+ET and US+MJ+ET samples after 72 h of storage by 53.4% and 86.6%, respectively, as compared with CT 0h samples. Based on the results herein obtained, the application of US can be an effective tool to enhance the extractability of certain glucocosinolate and pheniolic compounds in broccoli. Moreover, due to the synergistic effect observed on the accumulation of bioactive compounds, the combined application of US and MJ could be an effective approach to yield higher levels of glucosinolates and phenolic compounds in broccoli during storage. Additionally, recommendations for future investigations such as different US conditions, individual isothiocyanates analysis and analysis of gene expression were proposed in order to better understand the synergistic effect excerted by US and MJ on the accumulation of bioactive compounds in broccoli florets.
- Efecto del tratamiento con altas presiones hidrostáticas en la neurotoxicidad del ácido okadaico. Evaluación en cultivos de células granulares de cerebelo de rata Wistar.(2017-05-15) Hernández Ortega, César Augusto; Welti Chanes, Jorge; Fernández-Sánchez, María Teresa; Escobedo Avellaneda, Zamantha; Jorge Welti Chanes; Fernández-Sánchez, María Teresa; Escobedo Avellaneda, Zamantha; Chuck Hernández, Cristina; Novelli Ciotti, AntonelloExiste una gran diversidad de toxinas marinas que son producidas por diferentes tipos de algas. Dichas toxinas pueden acumularse dentro de moluscos y pescados cuya dieta es a base de algas. Esto representa un riesgo de salud para los consumidores de productos marinos ya que están expuestos a las toxinas que se han acumulado en la carne. El ácido okadaico (OKA) es una de las principales toxinas que se acumula en moluscos, y es responsable de ocasionar envenenamiento diarreico en humanos. El OKA es un fuerte inhibidor de proteínas fosfatasa, las cuales son responsables de controlar varios mecanismos moleculares dentro de las células. Si bien los síntomas son raramente letales, la contaminación de productos marinos con OKA ocasiona grandes pérdidas económicas. Debido a esto, es deseable buscar tratamientos que permitan detoxificar los moluscos. Lamentablemente tratamientos comunes en la industria alimentaria, tal como la cocción, no han sido capaces de eliminar el OKA de los moluscos. Las altas presiones hidrostáticas (APH) son una alternativa de tratamiento que podrían ser útiles para eliminar el OKA, con el beneficio adicionar de no alterar las propiedades sensoriales del molusco. En este trabajo se evaluó el efecto del tratamiento con APH en la toxicidad del OKA, considerando que en estudios previos se presentaron las primeras evidencias de que las APH pueden reducir la toxicidad del OKA. Se realizó el tratamiento con APH de soluciones de OKA a presión de 600 MPa durante 30 minutos, utilizando cuatro equipos de APH diferentes. La temperatura inicial del tratamiento fue de 25 ºC en todas las cosas, con excepción de un tratamiento único a temperatura inicial de 10 ºC. Se evaluó la toxicidad de las muestras de OKA (concentración 5 nM) con y sin tratamiento APH en cultivos primarios de neuronas de cerebelo de rata. Los resultados obtenidos indicaron que no existió diferencia significativa entre la neurotoxicidad del OKA sin tratamiento y la neurotoxicidad del OKA con tratamiento APH a 25 ºC. No obstante, se observó una reducción notable de la neurotoxicidad de muestras de OKA con tratamiento APH a 10 ºC. Esta disminución de la toxicidad a causa del tratamiento con APH se relacionó con una reducción de la activación de la apoptosis durante las primeras 24 horas de exposición de las células a la toxina, manteniendo el estado sano del núcleo celular y la estabilidad del ADN durante este periodo. Se concluyó que las APH a condiciones de 600 MPa y 30 minutos pueden reducir la neurotoxicidad del OKA cuando el tratamiento es realizado a temperaturas de refrigeración (10 ºC) pero no a temperaturas de nivel ambiente (25 ºC). Esta reducción de la toxicidad puede atribuirse a una disminución de la capacidad del OKA de interactuar con proteínas fosfatasas debido al tratamiento con APH, lo cual implica un posible cambio en el OKA que impida que la toxina interactúe correctamente con las fosfatasas.
- Studies on the growth of magnetospirillum gryphiswaldense under several conditions and its influence on the production of magnetosomes(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2017-05-12) Rodríguez Ceja, Jesús Gilberto; Aguilar Jiménez, Oscar Alejandro; Parra Saldivar, Roberto; Rostro Alanís, Magdalena de Jesús; Santacruz López, Yolanda ArletteIn recent years, Magnetospirillum gryphiswaldense has called attention because it is a bacteria capable of producing magnetic nanoparticles (magnetosomes) that can be used in a variety of applications, such as directed therapies for drug delivery. The production of magnetosomes has not reach threshole concentration to ensure feasible for industrial applications. Thus, it is of high importance to increase the magnetosome production. This research evaluated the effect of stirring rate (100, 200, 400 and 600 rpm) and hydrodinamic conditions based on Reynolds number for the grwoth and production of M. gryphiswaldense. Overall, the better results were obtained at 200 rpm, with correspond to higher biomass and oxygen transfer coeficient (KLA). Optimal impeler stirring rate was selected to grow bacteria, followed by an anaerobic stage with feeding, to induce the production of magnetosomes. The best magnetosome yield obtained was 18.79 mg/mL*day in a fed-batch culture, 11.78 in continuous culture and 1.4 in batch culture. Their respective specific growth (μ) and generation time (min) were 0.083 and 8.31, 0.022 and 31.38, 0.02 and 34.65. Three different pellets were obtained and their magnetosome content was extracted to be analyzed in Z-sizer, resulting in an average size of 100 nm. Also aggregates of magnetosomes were found. The findings reported here could serve as a basis for the future scaling up of a biological factory for nanocarriers for multiple research and/or medical applications.