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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Phylotranscriptomic Analysis of Pigment Biosynthesis in the Caryophyllales
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-12-02) Gutierrez Vences, Alma Yesenia; Rodríguez López, Carlos Eduardo; Chavez Santoscoy, Rocío; Dang, Thu-Thuy; Díaz de la Garza, Rocío; Treviñp, Manuel; School of Engineering and Sciences; Campus Monterrey
In plants, color is given by secondary metabolites, pigments, that absorb and emit color due to their chemical structure. Betalains are a class of tyrosine-derived pigments found exclusively in the Caryophyllales order, where they are concurrent with the anthocyanin pigments. These two types of pigments show a mutually exclusive pattern of expression rooted in a complex homoplastic distribution. Although the genetic events that led to the emergence of betalains in the Caryophyllales is known, the exact genetic evolutionary path that led to mutual exclusion remains elusive with multiple theories with a single or multiple emergences being proposed. This thesis combines orthology-based phylogenetic correlation analyses with comparative transcriptomics to investigate the molecular and evolutionary drivers of pigment biosynthesis, focusing on transcriptional regulation, pigment structural modification, and transport mechanisms. Co-expression analysis revealed potential regulatory co-option, suggesting that betalain-producing species may have repurposed ancestral anthocyanin regulatory mechanisms to drive their pigment emergence. Additionally, candidate enzymes such as UGT, SCPL, and MT were identified as key players in pigment structural modification and stabilization in betalain-producing species. The research also presents novel insights into the role of cellular transport mechanisms in pigment exclusion. Genes in orthogroups with statistically different occupancy annotated in processes related to vesicle trafficking were found to coexpress with biosynthetic modules, suggesting that transport may contribute significantly to the mutual exclusion of anthocyanins and betalains. These findings hint at a possible loss and co-option of anthocyanin transport mechanisms for betalain sequestration in vacuoles, raising questions about how shifts in transport specificity may drive pigment divergence.
Development of a multi-component disjointed tissue culture system using three-dimensionally printed polymeric scaffolds and microfluidic pumping
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-12) Romero Zepeda, Claudia Alejandra; Lozano Sánchez, Luis Marcelo; emipsanchez; Perfecto Avalos, Yacanxóchitl; García González, Alejandro; García Varela, Rebeca; School of Engineering and Sciences; Campus León; Chaires Oria, Jorge Isaac
In-vitro cellular culture plays a crucial role in preclinical research. While cost-effective, the pre- vailing 2D culture approach falls short in simulating realistic cellular interactions when these cells are grown in different but interacting spaces. Organs-on-a-Chip (OoC) devices have been developed to address this limitation, creating controlled micro-environments that mimic in-vivo tissue interaction conditions. This research addressed designing and assessing a microfluidic chip device based on ad- ditive manufacturing to analyze fibroblast and monocyte cell interaction grown in a separate culture apparatus. The OoC devices were created using Computer-Aided Design (CAD), and additive manu- facturing strategies using translucent resin as constructive material. The developed chip consisted of 200 mm2 cell culture area, a glass window for monitoring, and two inlets and outlets for fluid transfer and sampling. An instrumented peristaltic micro-pumping system induces fluid motion through the tubing that connects the manufactured microchips. Here, we show the ability of the developed 3D printed system to culture different cell lines, allow treatment addition without disturbing the system, and connect with a continuous flow between the devices without generating detectable cellular stress by enzymatic quantification. Finally, the interconnected system communicates between fibroblast and monocyte cultures by connecting two chips with micropumps through microscopic and cellular stress markers in selected cell lines. This results in a prototype for a multi-organ-on-a-chip-like device.
Analysis of cellular senescence in Spinocerebellar ataxia type 7 using bioinformatics and an inducible cell model
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-06-15) Ruiz Esparza Palacios, Vanessa; GONZALEZ MELJEM, JOSE MARIO; 316764; González Meljem, José Mario; puemcuervo; Pérez Méndez, Óscar Armando; García Aguirre, Ian Alaín; School of Engineering and Sciences; Campus Monterrey; Magaña Aguirre, Jonathan Javier
Spinocerebellar ataxia type 7 (SCA7) is an autosomal dominant inherited disorder manifested by the inability to coordinate balance, gait, and speech. Some experimental evidence suggests that nuclear inclusion of mutant ataxin-7 (ATX7) is part of the molecular basis of this disease and the origin of oxidative stress. In this thesis, we made a bioinformatic analysis that supports the hypothesis that SCA7 could share some mechanisms that are core features of senescent cells. Senescence is a permanent state of cell-cycle arrest, and core features of this phenotype include the expression of anti-proliferative molecules, the activation of a chronic DNA damage response, altered metabolic rates, and many others. We sought to establish a senescence induction protocol in a human fibroblast cell line as a positive control of senescence and found that induction with H2O2 at 1200 μM resulted in 79% of positive SA-β-gal cells. Afterwards, we were able to validate an inducible cell model of MIO-M1 cells, which were stably transduced to express a mutant ATXN7 gene carrying 64 CAG repeats and 10 CAG repeats. Administration of doxycycline (dox) induced the expression of the mutated protein causing the formation of nuclear aggregates. Furthermore, in a preliminary SA-β-gal assay, we found activity of this enzyme in 64 CAG cells, suggesting the presence of senescence features after the induction of the mutated protein. Based on our findings, we propose that the oxidative stress generated by the accumulation of the mutated protein could be leading to a senescence phenotype. Future evaluation of the Senescence-associated secretory phenotype (SASP) and markers of DNA damage will bring better understanding of the possible role of senescence in SCA7.
Characterization of the effect of UV-A light and agitation on the exopolysaccharide production of Chlorella vulgaris and Porphyridium cruentum
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-05) Garza Rodríguez, Zaida Berenice; GARZA RODRIGUEZ, ZAIDA BERENICE; 838147; Benavides Lozano, Jorge Alejandro; emipsanchez; Jacobo Velázquez, Daniel Alberto; Santacruz López, Yolanda Arlette; School of Engineering and Sciences; Campus Monterrey; Hernández Pérez, Jesús
Microalgae species are photosynthetic microorganisms that are a sustainable source of bioproducts due to their ability to reduce anthropogenic carbon dioxide in the atmosphere. The number of studies focused on their biologically active molecules, such as lipids, proteins, polysaccharides, and pigments, has been increasing in the last years due to their promising application as valuable products. In this context, the exopolysaccharides (EPS) from microalgal sources stand out as high-value molecules for their potential applications in the nutraceutical and pharmaceutical industries. However, studies aimed to find strategies and optimal conditions to promote the biosynthesis of EPS are still required to make these molecules economically feasible. The effect of ultraviolet light A (UV-A) is studied on biomass and EPS productivity of the red microalgae Porphyridium cruentum and the green microalgae Chlorella vulgaris testing three levels: L0, L1 and L2. Likewise, the effect of the agitation factor on cell growth and EPS productivity are analyzed for the two microalgae species testing two levels: A0 and A1. After recovering the EPS using centrifugation and diafiltration, their potential antioxidant activity was tested using the 2,2-Diphenyl-1-prykylhydrazyl (DPPH) radical scavenging assay. The UV-A light (315-400 nm), along with photosynthetic active radiation (PAR) increased EPS productivity in both C. vulgaris and P. cruentum to 1.21-fold and 2.43-fold, respectively, compared to light control conditions (PAR at 35.6 µmol m⁻² s⁻ ¹). The highest P. cruentum EPS productivity was 8.67 mg/g DW biomass at the highest tested UV-A level and lowest agitation(L2A0), and the highest biomass concentration was 8.57 g/L at the highest agitation (A1) due to a possible improvement in nutrient distribution. For C. vulgaris, the highest productivity was 1380 mg/g DW biomass at intermediate UV-A light level (L1). The highest biomass concentration of C. vulgaris was 0.34 g/L at the highest UV-A level (L2), treatment that conversely displayed the lowest EPS productivity (73.08 mg/g DW biomass). The study of antioxidant activity revealed that EPS show DPPH radical scavenging activity. The mean highest radical scavenging effect (%) of P. cruentum and C. vulgaris EPS was 56.48 ± 4.46 and 46.31 ± 3.2 at 4 mg/mL and 2 mg/mL, respectively. This work contributes to the characterization of abiotic factors that could potentially influence the production of high-value EPS molecules that display bioactivity of interest for health applications.
Anticancer activity of Elaphoglossum paleaceum (Hook. & Grev.) sledge derived phloroglucinols in human hepatocellular carcinoma cells through potential transcriptional modulation of microRNAs
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2022-01-05) Ruiz Manriquez, Luis Mario; PAUL, SUJAY; 512948; Paul, Sujay; emijzarate/puemcuervo; Arvizu Espinosa, María Goretti; School of Engineering and Sciences; Campus Monterrey; Sharma, Ashutosh
Phloroglucinols derivatives are an important class of secondary metabolites widely distributed in Pteridophytes of the genera Elaphoglossum that exhibit multiple biological activities, including anticancer activity. Recent evidence suggests that many phytochemicals, including polyphenols, can significantly alter microRNA (miRNAs) expression profiles. miRNAs are short non-coding RNAs that can act as tumor suppressors or onco-miRNAs regulating the expression of cancer-associated genes post-transcriptionally, therefore, modulating cancer progression. Elaphoglossum paleaceum (Hook. & Grev.) Sledge phloroglucinols derivatives have demonstrated cytotoxic activity against several cancer models. Nevertheless, its relative mechanism of action and its effect on HCC cells have not still investigated. The present study aimed to evaluate the anticancer activity of hexanic/chloroformic extract of E. paleaceum and investigate its mechanism(s) of action on tumor suppressor and onco-miRNAs expression involved in HCC carcinogenesis. E. paleaceum hexanic/chloroformic extract exhibited cytotoxicity against HepG2 cells. Moreover, an E. paleaceum hexanic/chloroformic extract concentration approximating the IC50 induced upregulation of tumor suppressors miR-200b-5p, miR-34-5p, Let-7b-5p, and miR-218b-5p. These modulatory effects might contribute to inhibiting HepG2 cell growth observed after treatment with E. paleaceum hexanic/chloroformic extract. These data demonstrate the ability of E. paleaceum phloroglucinols derivatives to impede hepatocellular carcinoma cell proliferation, possibly by modulating critical miRNAs and consequently their downstream targets, suggesting a novel mechanism of the anticancer potential of phloroglucinols derivatives and positioning them as potential novel anticancer agents.
Bioinformatic analysis of the expression pattern of an intron-retaining isoform of the Agrp transcript in arcuate nucleus neurons of mice
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-12-07) Gómez Montalvo, Jesús; GONZALEZ MELJEM, JOSE MARIO; 316764; González Meljem, José Mario; puemcuervo, emipsanchez; Treviño Alvarado, Víctor Manuel; de Obeso Fernández del Valle, Álvaro; School of Engineering and Sciences; Campus Monterrey; Avendaño Vázquez, Selma Eréndira
Known to be the regulatory center of hunger and satiety, the arcuate nucleus (ARC) harbors two neuronal populations that exert opposite effects on the regulation of food intake. Namely, AgRP and POMC neurons respectively trigger hunger and satiety. Although both neuronal groups are born within the same time interval in the developing hypothalamus of mouse, their peptidergic identities are established at distinct developmental timepoints. While the POMC identity is established as early as the embryonic period, the complete maturation of the AgRP peptidergic identity extends to the postnatal period. Previously, through RT-PCR, our group detected the presence of an Agrp transcript isoform that retained introns 3 and 4 (Agrp-i3,4) in the early postnatal, but not in the adult mouse hypothalamus. In this thesis project, the expression pattern of Agrp-i3,4 is analyzed in public RNA-seq datasets from ARC neurons of mice at different postnatal developmental stages. To identify intron retention events, iREAD was used, and to quantify the proportion of Agrp transcripts that retained introns 3 and 4, ASpli was employed. Using this bioinformatics approach, the largest proportion of Agrp-i3,4 was detected in ARC samples of P12 mice and there was a trend towards decreased retention of Agrp introns 3 and 4 at later developmental stages. Agrp-i3,4 was detected in poly-A RNA extracted from whole AgRP neurons, but not in the ribosomal fraction. On the other hand, food deprivation appeared to exert distinct effects on the proportion of the Agrp-i3,4 transcript depending on the duration of the fast. While in weaned mice fasted for 16 hours Agrp-i3,4 showed a slight increase, in adult mice fasted for 38 hours Agrp-i3,4 appeared to decrease. Unlike fasting, leptin treatment did not exert any effect on the retention of Agrp introns 3 and 4. Of note, IR was found in Agrp but not in other genes that characterize the ARC, such as Npy or Pomc. Taken together, the results presented in this thesis suggest that increased IR of the Agrp transcript may correlate with a lower maturation degree of the ARC during a time interval in which the AgRP peptidergic identity has not yet been fully established.
Evaluation of the protective effects of Time-Caloric Restriction in liver fibrosis in a DEN model
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-12-07) Arriaga González, Fernanda Guadalupe; ; De Donato Capote, Marcos; puemcuervo/tolmquevedo; Robles Espinoza, Carla Daniela; School of Engineering and Sciences; Campus Monterrey; Molina Aguilar, Christian
Liver fibrosis is, in most cases, the first stage of chronic liver disease. Advanced stages of fibrosis can evolve into cirrhosis and finally trigger hepatocellular carcinoma (HCC). The liver is the central metabolic organ of the body and diet is considered one of the main environmental factors that can impact its function. Most of the research up to this date centers on its effects in the last stages of this disease, meaning cirrhosis and HCC. Given that fibrosis is a reversible stage of liver damage, a better understanding of the effects of caloric restriction in this initial damage can help to prevent the development of advanced chronic liver disease. The aim of this study is to evaluate the possible protective effects of the time-caloric restriction protocol in the progression of liver fibrosis. Forty male Wistar rats were randomly assigned to four experimental groups according the food intake regime and treatment: AL, AL+DEN, T-CR, and T-CR+DEN. The AL groups had unrestricted access to food. T-CR groups had food access limited to a 2-hour period daily (from 11am to 13:00 pm). One group of each food intake regime receive a weekly intraperitoneal injection of diethylnitrosamine (DEN) of 50mg/kg to induce liver fibrosis. All rats were sacrificed after 10 weeks and blood samples were analyzed by standard spectrophotometric techniques to evaluate the nutritional state of the animals, liver damage, and the effects of T-CR on the inflammatory response. Histological evaluation of the livers was performed though Masson’s Trichrome and Hematoxylin-eosin staining. The T-CR regime prompted a ~40% reduction of daily food intake in comparison to the AL groups. However, the evaluation of biochemical parameters indicates that none of the groups was malnourished. There were no significant changes in liver chemistry results. However, histological evaluation revealed that the T-CR regime protected the liver from fibrogenic processes. Additionally, the T-CR groups showed a decrease in total leukocyte count when compared to the AL groups. The differential leukocyte count attributes this change to a decrease in lymphocyte population. Unexpectedly, an additional decrease in the lymphocyte count was observed in the groups with DEN treatment. It is difficult to posit a clear conclusion on the effect that the time-caloric restriction regime has on the inflammatory state of the animal models thus, further analysis has to be conducted.
In silico identification of cis-regulatory elements in folate biosynthesis and 1C metabolism genes in plants
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-11-26) Salinas Espinosa, Jessica Pamela; TREVIÑO ALVARADO, VICTOR MANUEL; 205076; Treviño Alvarado, Víctor Manuel; puemcuervo; Cuevas Díaz Durán, Raquel; Rodríguez López, Carlos; Martínez Ledesma, Juan Emmanuel; School of Engineering and Sciences; Campus Monterrey; Díaz de la Garza, Rocío Isabel
Folates (vitamin B9) are enzyme cofactors required for all organisms for one-carbon (1C) transfer reactions. A deficiency of these nutrients can lead to several health problems. Since humans are not natural producers of folates, the intake of these nutrients from plants is vital for human nutrition. Several techniques that involve the genetic modification of organisms have proved to be effective for the fortify plants with essential macronutrients. However, to achieve this, it is necessary to elucidate the metabolic control in plant systems. Although the genes involved in folate biosynthesis and 1C metabolism in plants are known, the mechanisms of transcriptional regulation have not yet been explored. This project focuses on discovering cis-regulatory DNA elements (motifs) using computational data analysis to provide insights regarding the regulation of folate biosynthesis in plants. For this, we first collected a compendium of known genes related to folate biosynthesis. Then, a database comprising the DNA promoter regions of folate biosynthesis and 1C metabolism genes in 19 different plant species was built and analyzed using different motif discovery algorithms. Afterward, the discovered motifs were tested for statistical significance and further associated with their putative biological role using other bioinformatics tools. A total of 149 statistically significant motifs (p < .05) were discovered in 18 of 19 species using the GimmeMotifs ensemble algorithm. These motifs were represented in 104 different regulatory networks built automatically from co-expression clusters obtained from each plant species. The results from this work could provide an insight into the transcriptional regulation of the folate biosynthesis pathway in plants. Furthermore, the elements found could be used for research in gene editing techniques to produce biofortified crops.
Culture of Euglena gracilis in photoautotrophy for paramylon production: effect of pH and media composition
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-11-15) Martín Roldán, María; ; Pacheco Moscoa, Adriana; puemcuervo/tolmquevedo; Senés Guerrero, Carolina; School of Engineering and Sciences; Campus Monterrey; Valledor González, Luis
Microalgae have been widely studied in biotechnology for their application in various areas such as bioremediation, production of biofuels, or use in nutrition. More specifically, certain species are recognized to produce compounds of high commercial value. Euglena is a group of microalgae characterized by the generation of the reserve polysaccharide paramylon, with promising prospects for its application in pharmacology, nutrition, to produce bioplastics, or biodiesel. Paramylon is accumulated mainly in the presence of organic carbon in the culture medium; however, this represents a challenge when establishing a large-scale culture due to the risk of biological contamination. In this study, an extensive study of the literature was carried out with respect to the autotrophic culture of Euglena gracilis destined to paramylon production. As a result, we evaluated the effect of culture pH, vitamin supplementation, and nitrogen source in the culture medium in order to reach the maximum biomass productivity. A pH of 7.5 and ammonium as nitrogen source were optimum for the autotrophic culture of E. gracilis, while an improvement in productivity was not observed with vitamin supplementation. Finally, it was possible to scale up the photoautotrophic culture of E. gracilis to a 1-L airlift photobioreactor. In conclusion, there is still much optimization work to achieve the biomass and paramylon productivity reported for the heterotrophic cultures of E. gracilis, but the results of this study reveal its viability.
Induction of antigen-independent T cell proliferation in vitro
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2021-06-11) Ureña Herrera, Milena; Brunck, Marion E.G.; puemcuervo; Vázquez Garza, Eduardo; Licona Cassani, Cuauhtémoc; School of Engineering and Sciences; Campus Monterrey
Innate immunity is the first line of defense of the body against pathogens and it responds in a nonspecific way to all threats. Some of its major responses are inflammation and antiviral defense, which overall help to prevent, control, and eliminate infections. It also allows the induction of an adaptive immune response, which is determinant to prevent re-infection. The adaptive immune response depends on antigen recognition, it triggers stimulation signals that In vivo induce T lymphocytes to proliferation response in addition to cytokine production and a modification to the pattern of expression of cell surface glycoproteins. T lymphocytes are one of the effector cells on the adaptive immune response. Their function of destroying the pathogens and their toxic molecules have made them popular for the development of cell therapies, for example, the CAR T cell therapies, which are based on modifying the antigen receptors of the T cells to allow them to detect and destroy cancer cells. Therefore, mimic in vitro as much as possible in vivo stimulation to induce cell proliferation is key to improve cell therapy manufacturing, decrease the time of production and costs. In this work, different stimulation protocols with PMA, ionomycin, IL-2, and Dynabeads™ were tested and evaluated in terms of proliferation induction, and progression in the cell cycle, changes in the expression of surface proteins, and gene transcription levels of relevant genes. A correct overexpression of activation markers, as well as cytokines transcription, is confirmed, however, the stimuli were not correct to induce a significant increase in cell proliferation, and contrary to expected, it seems that induced cells to apoptosis.

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