Universidad Nacional Autonoma de Mexico View Institution's Website 14 articles published in JoVE Environment Biogas Purification through the use of a Microalgae-Bacterial System in Semi-Industrial High Rate Algal Ponds Mariana Vega Blanes1, Isaac Jhonnatan Pérez-Hermosillo2, Arnold Ramírez Rueda1, Armando González Sánchez1 1Instituto de Ingeniería, Ciudad Universitaria, Universidad Nacional Autónoma de México, 2Granos y Servicios Integrales Air pollution impacts the quality of life of all organisms. Here, we describe the use of microalgae biotechnology for the treatment of biogas (simultaneous removal of carbon dioxide and hydrogen sulfide) and the production of biomethane through semi-industrial open high-rate algal ponds and subsequent analysis of treatment efficiency, pH, dissolved oxygen, and microalgae growth. Biochemistry Estimation of Structural Sensitivity of Intrinsically Disordered Regions in Response to Hyperosmotic Stress in Living Cells Using FRET Constanza Enriquez-Toledo*1, Cesar A. Ponce-Diego*1, Cesar L. Cuevas-Velazquez1 1Departamento de Bioquímica, Facultad de Química, Universidad Nacional Autónoma de México Intrinsically disordered regions (IDRs) are flexible protein domains that modify their conformation in response to environmental changes. Ensemble fluorescence resonance energy transfer (FRET) can estimate protein dimensions under different conditions. We present a FRET approach to assess IDR structural sensitivity in living Saccharomyces cerevisiae cells under hyperosmotic stress. Immunology and Infection A Strategy for the Study of IL-9-Producing Lymphoid Cells in the Nippostrongylus brasiliensis Infection Model Ofelia Muñoz-Paleta*1, Enrique Olguín-Martínez*1, Blanca Estela Ruiz-Medina*1, Alejandra Alonso-Quintana1, Ma Cleofas Marcial-Medina1, Paula Licona-Limón1 1Departamento de Biología Celular y del Desarrollo, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México IL-9-expressing T and ILC2 cells are induced during N. brasiliensis infection, yet their characterization has been largely overlooked in the infected intestine due to their low frequency and differential kinetics. This protocol describes the isolation of these cells from different target organs and confirmation of their identity via flow cytometry at different infection stages. Developmental Biology Chicken Recombinant Limbs Assay to Understand Morphogenesis, Patterning, and Early Steps in Cell Differentiation Jessica Cristina Marín-Llera1, Montse Fernández-Calderón2, Jesús Chimal-Monroy1 1Departamento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, 2Departamento de Anatomía y Biología Celular and IDIVAL, Facultad de Medicina, Universidad de Cantabria Recombinant limbs are a powerful experimental model that allows for studying the process of cell differentiation and the generation of patterns under the influence of embryonic signals. This protocol presents a detailed method for generating recombinant limbs with chicken limb-mesodermal cells, adaptable to other cell types obtained from different organisms. Medicine Isolated Lung Perfusion System in the Rabbit Model Alejandro Pacheco-Baltazar2, José Luis Arreola-Ramírez1, Jesús Alquicira-Mireles1, Patricia Segura-Medina1 1Departamento de Investigación en Hiperreactividad Bronquial, Instituto Nacional de Enfermedades Respiratorias, 2Facultad de Medicina, Universidad Nacional Autónoma de México The isolated rabbit lung preparation is a gold standard tool in pulmonary research. This publication aims to describe the technique as developed for the study of physiological and pathological mechanisms involved in airway reactivity, lung preservation, and preclinical research in lung transplantation and pulmonary edema. Behavior A Two-interval Forced-choice Task for Multisensory Comparisons Fabiola Duarte1, Tonatiuh Figueroa1, Luis Lemus1 1Neurociencia Cognitiva, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México Psychophysics is essential for studying perception phenomena through sensory information. Here we present a protocol to perform a two-interval forced-choice task as implemented in a previous report on human psychophysics where participants estimated the duration of visual, auditory, or audiovisual intervals of aperiodic trains of pulses. Cancer Research Analyzing the Communication Between Monocytes and Primary Breast Cancer Cells in an Extracellular Matrix Extract (ECME)-based Three-dimensional System Nancy Adriana Espinoza-Sánchez*1,2, Gloria Karina Chimal-Ramírez*1, Ezequiel Moisés Fuentes-Pananá1 1 Here, we describe a three-dimensional culture method to analyze the morphology of primary breast cancer cells, as well as to study their direct/indirect interactions with monocytes and the outcomes such as collagen degradation, immune cell recruitment, cell invasion, and promotion of cancer-related inflammation. Biology CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion Orlando Santillán1, Miguel A. Ramírez-Romero2, Guillermo Dávila3 1Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, 2Oncomedic Incorporation, Ciudad de México, 3Laboratorio Internacional de Investigación sobre el Genoma Humano, Universidad Nacional Autónoma de México Here, we present chimera assembly by plasmid recovery and restriction enzyme site insertion (CAPRRESI), a protocol based on the insertion of restriction enzyme sites into synonym DNA sequences and functional plasmid recovery. This protocol is a fast and low-cost method for fusing protein-coding genes. Biochemistry Defining Substrate Specificities for Lipase and Phospholipase Candidates Diana X. Sahonero-Canavesi1, Maritza Zavaleta-Pastor1, Lourdes Martínez-Aguilar1, Isabel M. López-Lara1, Otto Geiger1 1Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México Many predicted (phospho)lipases are poorly characterized with regard to their substrate specificities and physiological functions. Here we provide a protocol to optimize enzyme activities, search for natural substrates, and propose physiological functions for these enzymes. Biochemistry Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions Abril Gijsbers1, Takuya Nishigaki2, Nuria Sánchez-Puig1 1Departamento de Química de Biomacromoléculas, Instituto de Química, Universidad Nacional Autónoma de México, 2Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México Protein interactions are at the heart of a cell's function. Calorimetric and spectroscopic techniques are commonly used to characterize them. Here we describe fluorescence anisotropy as a tool to study the interaction between the protein mutated in the Shwachman-Diamond Syndrome (SBDS) and the Elongation factor-like 1 GTPase (EFL1). Developmental Biology Affinity Labeling Detection of Endogenous Receptors from Zebrafish Embryos Tonatiuh Molina-Villa1, Valentín Mendoza1, Fernando López-Casillas1 1Cell and Developmental Biology, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México A novel technique for the detection of low abundance endogenous receptors present in zebrafish embryos is described. We have named it AFLIP because it consists of affinity labeling of the receptor by its ligand linked to immunoprecipitation. Immunology and Infection Isolation of Viral Replication Compartment-enriched Sub-nuclear Fractions from Adenovirus-infected Normal Human Cells Paloma Hidalgo1,2, Ramón A. Gonzalez1 1Centro de Investigación en Dinámica Celular, Instituto de Investigación en Ciencias Básicas y Aplicadas, Universidad Autónoma del Estado de Morelos, 2Instituto de Biotecnologìa, Universidad Nacional Autónoma de México We provide a novel strategy to isolate viral replication compartments (RC) from adenovirus (Ad)-infected human cells. This approach represents a cell-free system that can help to elucidate the molecular mechanisms regulating viral genome replication and expression as well as regulation of viral-host interactions established at the RC. Biology Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging Esperanza Mata-Martínez1, Omar José1, Paulina Torres-Rodríguez1, Alejandra Solís-López1, Ana A. Sánchez-Tusie1, Yoloxochitl Sánchez-Guevara1, Marcela B. Treviño2, Claudia L. Treviño1 1Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 2Math and Sciences Department, Edison State College Intracellular Ca2+ dynamics are very important in sperm physiology and Ca2+-sensitive fluorescent dyes constitute a versatile tool to study them. Population experiments (fluorometry and stopped flow fluorometry) and single cell experiments (flow cytometry and single cell imaging) are used to track spatio-temporal [Ca2+] changes in human sperm cells. Immunology and Infection A Simple and Efficient Method to Detect Nuclear Factor Activation in Human Neutrophils by Flow Cytometry Erick García-García1, Eileen Uribe-Querol2, Carlos Rosales3 1Department of Biological Sciences, University of Alberta, 2División de Estudios de Posgrado e Investigación, Facultad de Odontología, Universidad Nacional Autónoma de México, 3Department of Immunology, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México Neutrophils are the most abundant leukocytes in blood. Neutrophils possess transcriptionally regulated functions such as production of proinflammatory cytokines and inhibition of apoptosis. These functions can be studied with the method presented here, which allows detection and quantification of nuclear factors by flow cytometry in isolated nuclei
