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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- Evaluation of three fluorescent lateral flow assay platforms for the detection of leptin, adiponectin, and insulin in mexican adults and pediatric populations(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2025-06-06) Aguilar Cavada, Magaly; González González, Mirna Alejandra; emipsanchez; Rito Palomares, Marco Antonio; Coale Willson, Richard; Escuela de Medicina y Ciencias de la Salud; Campus MonterreyObesity is a major public health concern on a global scale. In Mexico, the prevalence of obesity among adults was 36.9% in 2022, while rates among children and adolescents were 18.1% and 17.2%, respectively. The prevalence of adult obesity in Mexico ranks among the highest in the world and is closely associated with the development of chronic diseases and metabolic syndrome. While body mass index (BMI) remains the standard metric for obesity, it fails to reflect metabolic health accurately. Public health policies are moving towards overweight and obesity prevention during childhood to avoid complications later in life. Effective treatment and management of this disease requires novel monitoring tools to help improve the assessment of metabolic status and associated health risks. Leptin, adiponectin, and insulin are three protein hormones closely related to each other. They are involved in energy expenditure regulation, lipid and glucose metabolism, and immune response and inflammation. The altered levels of these proteins are linked to coronary disease, insulin resistance, metabolic syndrome, and cancer, all of which are related to the correct function of adipose tissue. Lateral flow assays (LFA) provide inexpensive, easy to use, and portable alternative to classical biomarker detection techniques. In collaboration between the Bioengineering and Medical Devices Unit of the Institute for Obesity Research at Tecnológico de Monterrey and Dr. Richard Willson’s team from the Department of Chemical and Biomolecular Engineering at the University of Houston, three fluorescence-based lateral flow assays have been developed for the detection of leptin, adiponectin, and insulin. This thesis aims to validate the performance of three LFA for the detection of leptin, adiponectin, and insulin by analyzing samples from Mexican adult and pediatric participants and comparing the results to concentration values obtained through ELISA.
- Development and refinement of an aqueous two-phase system three-dimensional cell culture method for MDA-MB-231 cancer cell spheroid analysis(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-06-04) Villarreal Alejandre, Javier; González González, Mirna Alejandra; emimmayorquin; Benavides Lozano, Jorge Alejandro; Willson, Richard Coale; Escuela de Medicina y Ciencias de la Salud; Campus Monterrey; Rito Palomares, Marco AntonioAqueous two-phase systems (ATPS) are generated by mixing immiscible solutions of polymer-polymer combinations above certain concentrations forming two phases, allowing selective partition of biological particles, such as cells. Leveraging this, novel biomedical applications have emerged using ATPS for three-dimensional (3D) cell culture due to their ability to recapitulate the tumor in vivo conditions. Current approaches require sophisticated equipment and highly trained personnel for their production. To address this, a 3D cell culture method using ATPS and basic laboratory equipment for culturing cancer cell spheroids (CCS) has been designed. Nevertheless, to produce CCS of well-defined baseline characteristics with this approach, refinement of culture conditions is necessary for accurate drug-response evaluations. CCS configuration is achieved through the confinement of cells within droplets of 6.4% w/w dextran (DEX) 500,000 g/mol that are subsequently submerged in 5% w/w polyethylene glycol (PEG) 35,000 g/mol. Confining cells in DEX at different cell densities (5.0x103 -1.6x104 cells) produced CCS of diameters ranging from 419.51±64.16 μm to 818.11±83.27 μm after 48 h of culture, indicating that CCS of increasing size are achieved. Furthermore, 8-day examination of CCS viability showed reduced cell metabolic activity compared to their 2D culture (p-value <0.05) and consistent viability onwards day 4 of culture (p-value >0.05), reflecting diminished diffusion of nutrients and enabling assessing long-term biological processes. Finally, CCS showed a deviation from a sigmoidal response to paclitaxel (PTX), displaying an increase of 31% in maximum cell viability inhibition (Emax) in contrast with their 2D counterpart, signifying drug resistance. In conjunction, these results indicate that the ATPS-3D approach can produce in vitro models of consistent, predictable signals for end-point assays and biologically complex responses to drugs.
- Assessment of non-invasive colorimetric methods for pH and glucose determination in human saliva(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-05) Flores de la Toba, Raquel Fernanda; González González, Mirna Alejandra; emimmayorquin; Benavides Lozano, Jorge Alejandro; Willson, Richard Coale; Escuela de Medicina y Ciencias de la Salud; Campus Monterrey; Rito Palomares, Marco AntonioMonitoring glycemic biomarkers such as glucose is essential for the management of metabolic diseases such as obesity and its associated comorbidities. Traditional methods for glucose monitoring are based on invasive blood tests, using other biological fluids like saliva could be an alternative. Point-Of-Care Tests (POCT) are widely used for saliva analysis using colorimetric enzyme-based methods. However, the influence of pH on enzymatic activity is often overlooked in the development of such analytical devices. This study aims to assess the effectiveness of non-invasive colorimetric methods for pH determination, and glucose quantification by microfluidic paper-based analytical devices (μPADs) for the analysis of human saliva samples. Colorimetric methods were developed for pH determination using spectrophotometry with bromothymol blue, image analysis of the color channel of test strips using a smartphone, and adaptation of this method to an App. The effect of pH on (μPADs) for glucose quantification was analyzed in buffer solutions, pH determination methods and the μPAD for glucose quantification were validated in saliva samples from 29 healthy subjects. Test strips and the App performed better than spectrophotometry when tested in saliva, yielding a mean of pH 7.58. NaOH was selected as the wet etching agent in the fabrication of the μPAD for glucose quantification (LOD of 0.14 mM), at a pH 6 device’s response was significantly affected. Although no correlation was found between pH and salivary glucose, pH may provide insight into oral health. In addition, a moderate positive correlation (r=0.53, p-value=0.0054) was found between salivary glucose and body mass index (BMI), suggesting a potential application in metabolic monitoring. Implementing colorimetric methods (pH and glucose quantification) could enhance saliva analysis tools for POCT and metabolic monitoring. Therefore, it is proposed that future implementation of these methods will serve as a useful auxiliary tool for point-of-care testing, such as glycemic biomarker detection.
- Development, optimization and evaluation of a microfluidic, paper-based, analytical device for glucose and uric acid detection: a proof-of-concept(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2023-06-01) González Torres, Luis Fernando; González González, Mirna Alejandra; puemcuervo, emimayorquin; Zavala Arcos, Judith; Aguilar González, Cristóbal Noé; School of Engineering and Sciences; Campus Monterrey; Ortíz Martínez, MargaritaGlucose and uric acid are systemic metabolites of clinical interest in patients with obesity. Glucose plays a crucial role in multiple metabolic processes and is an indicator of pathological conditions in obesity-related diseases such as diabetes. Additionally, untreated hyperuricemia, defined as serum uric acid levels exceeding 400 µM, is associated with obesity and can lead to the development of conditions such as gout and kidney dysfunction. Therefore, regular testing of glucose and uric acid levels can be relevant as preventive measures for screening and monitoring purposes. Within the field of point-of-care testing, microfluidic, paper-based analytical devices offer several advantages, including compact size, portability, affordability, biocompatibility, and simplicity. Enzymatic colorimetry serves as an effective detection method due to its low cost, rapid response time, and high selectivity. Furthermore, no complex equipment is required, and a smartphone can be employed to capture the color response for analysis in ImageJ. As a microfluidic pattern generation method, selective wet etching stands out given its fast, simple, and inexpensive procedure. In this work, a microfluidic, paper-based, colorimetric device is developed to detect glucose and uric acid in solution. Trimethoxy(octyl)silane was used to turn the surface of the paper hydrophobic, and NaOH was used as the etching agent to generate the hydrophilic pattern. The detection limits for glucose and uric acid were 0.02 and 0.04 mM, respectively. Additionally, the glucose and uric acid assays exhibit linear responses encompassing the physiological range found in saliva and serum. In the future, this device could be further adapted and validated for the detection of glucose and uric acid in relevant biofluids.
- 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.