This protocol details a method to analyze two tissue samples at the transcriptomic level using microarray analysis, ingenuity pathway analysis (IPA) and gene set enrichment analysis (GSEA). Methods such as these provide insight into the mechanisms underlying biological differences across two samples and thus can be applied to interrogate a variety of processes across different tissue samples, conditions, and the like. The full method detailed below can be applied to determine the effects of muscle-specific Notch1 activation in the mouse model and to analyze previously published microarray data of human liposarcoma cell lines.
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http://dx.doi.org/10.21769/BioProtoc.2999 | DOI Listing |
ACS Biomater Sci Eng
January 2025
Mechanical Engineering Department, Worcester Polytechnic Institute, Worcester, Massachusetts 01609, United States.
Mechanical properties of engineered connective tissues are critical for their success, yet modern sensors that measure physical qualities of tissues for quality control are invasive and destructive. The goal of this work was to develop a noncontact, nondestructive method to measure mechanical attributes of engineered skin substitutes during production without disturbing the sterile culture packaging. We optimized a digital holographic vibrometry (DHV) system to measure the mechanical behavior of Apligraf living cellular skin substitute through the clear packaging in multiple conditions: resting on solid agar as when the tissue is shipped, on liquid media in which it is grown, and freely suspended in air as occurs when the media is removed for feeding.
View Article and Find Full Text PDFInt Endod J
January 2025
Department of Restorative Dentistry-Endodontics, Piracicaba Dental School, Universidade Estadual de Campinas, Piracicaba, São Paulo, Brazil.
Aim: Calcium silicate-based cements have been widely used in dentistry mainly due to their physicochemical and biological properties. Commercially available materials use radiopacifiers containing metals (bismuth, tantalum, tungsten and/or zirconium). To investigate volumetric changes, in vivo biocompatibility and systemic migration from eight commercially available materials, including powder/liquid and 'ready-to-use' presentations.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Bioscience, Research and Early Development, Oncology, AstraZeneca, Cambridge, Cambridgeshire, UK.
A protocol for the preparation of tissue extracts for the targeted analysis ca. 150 polar metabolites, including those involved in central carbon metabolism, is described, using a reversed phase ion pair U(H)PLC-MS method. Data collection enabled in high-resolution mass spectrometry detection provides highly specific and sensitive acquisition of metabolic intermediates with wide range physicochemical properties and pathway coverage.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Division of Metabolomics, Medical Research Center for High Depth Omics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
Lipidomics has attracted attention in the discovery of unknown biomolecules and for capturing the changes in metabolism caused by genetic and environmental factors in an unbiased manner. However, obtaining reliable lipidomics data, including structural diversity and quantification data, is still challenging. Supercritical fluid chromatography (SFC) is a suitable technique for separating lipid molecules with high throughput and separation efficiency.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Division of Systems Medicine, Department of Metabolism Department of Metabolism, Digestion and Reproduction, Imperial College, London, UK.
Metabolic profiling performed using untargeted metabolomics of different, complex biological samples aims to apply agnostic/holistic, hypothesis-free, analysis of the small molecules that are present in the analyzed sample. This approach has been the center of major investments and dedicated efforts from the research community for many years. However, limitations and challenges remain, particularly with regard to the validation and the quality of the obtained results.
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