Ciencia e Ingeniería en Alimentos y Biotecnología
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Item Análisis de variantes de splicing y factores de transcripción (HSF1, CBP y Sp1) en la evolución de la enfermedad de Huntington(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) Silva Gamboa, Christopher Joel; Galarza Galarza, Cristian FernandoHuntington's disease (HD) is a neurodegenerative disorder characterized by the expansion of CAG repeats in the HTT (Huntingtin) gene, leading to significant molecular dysfunctions. In the present study, the impact of splicing variants (SV) and transcription factors (TF) on disease progression was investigated using an in-silico approach with sequencing data based on RNA-seq technologies. Differential splicing variants were identified and their possible influence on the generation of aberrant HTT gene isoforms was analysed. Among these variants, the 109CAG condition showed a particularly severe impact on mRNA processing, primarily affecting huntingtin protein stability. In addition, the functional networks of the HSF1, CBP, and Sp1 factors were constructed and analysed. CBP and Sp1 emerged as central nodes, indicating that they play key roles in global epigenetic and genetic regulation, while HSF1 does not show significant connections with the others. These results suggest that interactions between SVs and TFs exacerbate cellular dysfunction, contributing to HD progression. Finally, the research concludes that both splicing variants and transcription factors represent critical components in HD pathogenesis. These findings provide a solid basis for exploring therapeutic interventions aimed at modulating these molecular processes.Item Evaluación in silico de microbiomas intestinales para identificar biomarcadores específicos en enfermedades de Crohn y Celíaca usando técnicas de metagenómica comparativa(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) Lucero Chisaguano, Israel Joel; Galarza Galarza, Cristian FernandoUnderstanding the intestinal microbiota is crucial for advancing the diagnosis and treatment of inflammatory diseases such as Crohn's disease (CDr) and Celiac disease (CD). This study employs comparative metagenomics techniques to identify specific biomarkers to differentiate these pathologies. The results highlight the relevance of microbial dysbiosis as a common factor, but also highlight significant differences in the composition and functions of the intestinal microbiota in each disease, which will advance the understanding of these pathologies, and create opportunities for the development of diagnostic tools and innovative therapies. The analysis was performed using 16S rRNA gene samples, processed with the QIIME2 tool under strict sample quality criteria. In parallel, the impact of sex was evaluated by analyzing significant variations in beta microbial diversity in patients with CDr. Functional analysis performed with PICRUSt revealed specific metabolic pathways related to inflammation in CD and oxidative stress in rCD. Microbial differences observed showed an increase in Proteobacteria, along with reductions in Firmicutes and Actinobacteria. In addition, biomarkers were identified in common such as, Faecalibacterium and Lachnospiraceae, and specific ones such as Alistipes in CDr and Lachnoanaerobaculum in CD. These results validate metagenomics as a tool to explore the microbiome-disease relationship, facilitating early diagnosis and the design of personalized therapies aimed at restoring microbial balance, marking a significant milestone in the development of precision medicine for inflammatory bowel diseases.Item Evaluación in silico de microbiomas intestinales para identificar biomarcadores específicos en enfermedades de Crohn y Celíaca usando técnicas de metagenómica comparativa(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) López Herrera, Alan Joshua; Galarza Galarza, Cristian FernandoUnderstanding the intestinal microbiota is crucial for advancing the diagnosis and treatment of inflammatory diseases such as Crohn's disease (CDr) and Celiac disease (CD). This study employs comparative metagenomics techniques to identify specific biomarkers to differentiate these pathologies. The results highlight the relevance of microbial dysbiosis as a common factor, but also highlight significant differences in the composition and functions of the intestinal microbiota in each disease, which will advance the understanding of these pathologies, and create opportunities for the development of diagnostic tools and innovative therapies. The analysis was performed using 16S rRNA gene samples, processed with the QIIME2 tool under strict sample quality criteria. In parallel, the impact of sex was evaluated by analyzing significant variations in beta microbial diversity in patients with CDr. Functional analysis performed with PICRUSt revealed specific metabolic pathways related to inflammation in CD and oxidative stress in rCD. Microbial differences observed showed an increase in Proteobacteria, along with reductions in Firmicutes and Actinobacteria. In addition, biomarkers were identified in common such as, Faecalibacterium and Lachnospiraceae, and specific ones such as Alistipes in CDr and Lachnoanaerobaculum in CD. These results validate metagenomics as a tool to explore the microbiome-disease relationship, facilitating early diagnosis and the design of personalized therapies aimed at restoring microbial balance, marking a significant milestone in the development of precision medicine for inflammatory bowel diseases.Item Evaluación in silico de la influencia de la heterogeneidad tumoral en la evolución clonal del cáncer de mama(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) Caicedo Albán, Samantha Lizbeth; Galarza Galarza, Cristian FernandoThe present study seeks to understand the influence of tumor heterogeneity on cancer progression and metastasis development, proposing a starting point for personalized treatment. Data from breast cancer were used, with sequences from samples belonging to primary tumors and tumors in the lymph nodes. Bioinformatics techniques were applied for processing, and differential expression analysis was used to identify genes that were expressed in each of the experimental conditions. Interaction networks were represented to identify the different interactions of elite genes with other genes. The functionality of these genes was validated using the functional enrichment technique to identify the role that these genes play in cell integrity and homeostasis. A search for variants was performed with the selected genes to identify the influence that these genes have on the development and progression of metastasis. Tumor heterogeneity on clonal evolution was evaluated by analyzing the influence of the variants of each type of tumor, observing that GL has a higher incidence on metastasis compared to TP.Item Estandarización del método PMAxxTM qPCR para la cuantificación de la carga viable de Salmonella spp. en contenido cecal de origen avícola(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2022-03) Corrales Martínez, Joselyn Griselda; Calero Cáceres, William RicardoCurrently, the standardization and validation of quantitative molecular techniques such as qPCR are important for the quality assurance and research sectors. Especially, the standardization of qPCR protocols for the detection of Salmonella spp. is essential, due to the impact it has on the health of the population. The present work aimed to standardize the PMAxxTM-based qPCR method for the detection and quantification of viable cells of S. infantis in poultry cecal content samples. First, staining with PMAxx bacterial viability stain (Trademark) of pure Salmonella culture samples was performed to inhibit the signal of non-viable cells. After DNA extraction with two different kits, a bank of serial dilutions was made, which were used for the construction of standard curves using the Illumina Eco Thermal Cycler (Trademark) Real-Time PCR System. In the analysis of the results, the concentration and volume of dye used significantly inhibited the signal of dead cells. The standard curves obtained with the two DNA extraction kits showed a high linearity greater than 0.99 and a limit of quantification of Ct 31.4. However, the best efficiency was obtained with the QIAamp Fast DNA Stool Mini Kit, which was 84.99 percent. Therefore, the optimization of the method obtained with this kit was used because it had acceptable yield values for the subsequent quantification of S. infantis in samples of poultry cecal contents.Item Validación de técnicas de PCR cuantitativa (qPCR) para la cuantificación de genes de resistencia a antibióticos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2019-12) Valle Ramos, María Judith; Calero Cáceres, William RicardoThe increment in the usage of genomic techniques applied to diagnosis and research demands to have validated techniques, in order to obtain reliable and reproducible results during the experimental tests. This work aimed to validate real-time PCR (qPCR) techniques, using both TaqMan and Sybr Green methods. The applied equipment was 7500 Fast Real-Time PCR System thermal cycler. Pure standards of the different recombinant plasmids (blaTEM, sul1, qnrS, ermB, and 16S rDNA) were used. Additionally, a synthetic plasmid standard was designed for tetW gene. The results indicate that the standard curves presented high efficiencies (between 97.4 to 109. 8 percent), as well as linear correlations greater than 0.99 and limits of quantification between Ct 34.4 to 34.8. Therefore, it was demonstrated that the qPCR techniques have acceptable performances since the values obtained are within the optimal ranges. These validated qPCR techniques will be used as a control tool to evaluate the dissemination of antibiotic resistance genes from environmental samples, obtaining truthful data in a short time.Item Identificación y clasificación de bacterias con potencial en biotecnología vegetal(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-12) Reinoso Caicedo, Gladys Estefanía; Rodríguez Meza, Carlos AlbertoForty-four bacteria were classified, which were separated into 3 groups of interest according to their possible uses in plant biotechnology processes. In the phenotypic study of the strains, data were obtained from their macroscopic, microscopic and physiological characteristics. By direct observation of the colony, stains, tests of resistance to different conditions of the medium and tests of use of different sources of carbon. In the molecular study the characteristics were obtained by the BOX PCR reaction of all the strains. Both, phenotypic and molecular characteristics were analyzed using the coefficient of Simple similarity (SSM) and the UPGMA algorithm. The species groups formed in this analysis helped to select the strains that were identified by sequencing the 16s rDNA gene. To obtain the species that belonged each strain was carried out the phylogenetic analysis in the program PHYDIT, using the coefficient of Jukes-Cantor and Neighbour joining algorithm to obtain the phylogenetic tree. From which a total of 16 different species were obtained, where those belonging to the genus Bacillus were the dominant ones with a total of 11 species. Two species of Enterobacter were also obtained and only one species of Rhizobium, Pseudomonas and Lysinibacillus. It should be emphasized that these species are distributed in the 3 groups of interest demonstrating the diversity existing in the collection of bacteria used in this study.Item Derarrollo de herramientas de biología sintética para la bacteria no modelo Burkholderia sacchari: un nueva plataforma para una producción más limpia(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2018-11) Barreno Sánchez, María José; Pérez Betancourt, YunysThe metabolic diversity in microorganisms can provide the basis to create new biochemical products. However, most metabolic engineering projects use a handful of established model organisms and, therefore, a challenge to take advantage of the potential of novel microbial functions is the ability to express new genes or directly use non-model organisms. Genetic manipulation of non-model microorganisms is still a challenge due to the specific nuances of the organism that make universal molecular genetic tools difficult. However, in recent years, unprecedented advances have been made in synthetic biology and the development of molecular genetic tools. That is why the objective of this study is to develop useful tools of synthetic biology to enhance the use of the non-model bacterium Burkholderia sacchari, as an alternative to biological production, since it has been proven that it can synthesize several high-value products using different sources of carbon. In this work we used the BglBrix plasmids, a standard that includes all the basic biological parts necessary for a vector to be efficient, stable and fulfill its function within the cell. The transformation of Burkholderia sacchari cells with the BglBrix plasmids was carried out by chemical method using RbCl, obtaining high transformation efficiency. Dose-induction assays were performed to evaluate the expression of fluorescent proteins at a maximum concentration of inducer.Item Determinación molecular de la biodiversidad de levaduras asociada al Canal de riego Latacunga - Salcedo - Ambato (CRLSA).(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2016-05) Hidalgo Escobar, Edith Paola; Pérez Betancourt, YunysThe Latacunga-Salcedo-Ambato irrigation channel (LSAIC) is an important environment for agriculture located in the central region of the country. About 24000 ha of crops are irrigated with this water. However, the channel receives large flows of pollutants that result from the neighboring industrial activity. To determine the diversity of yeast within this habitat, seven sampling points were selected along the 36 km comprising the channel. From the samples of water taken, 13 yeast species belonging to 10 genera were isolated. Phylogenetic analysis showed a heterogeneous population, where 77% of the population belongs to the phylum Ascomycota compared with 23% corresponding to the phylum Basidiomycota. Moreover, the molecular diversity found in LSAIC was compared with other polluted waters environments (River Santiago Argentina, River Danube Bratislava, River Doce Brazil) with similar physicochemical characteristics. It was determined that most of the yeast population was different.