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Will canopy nitrogen subscriber base boost carbon dioxide sequestration simply by bushes?
A green and sustainable nitrone formation reaction via visible-light-promoted reaction of aryl diazoacetates with nitrosoarenes is described. This protocol exhibits good functional group tolerance and broad substrate scope for both aryl diazoacetates with nitrosoarenes. Comparing the reported methods for the synthesis of nitrones from nitrosoarenes, the reaction described herein occurs under sole visible-light irradiation without the need of any catalysts and additives.This chapter aims to provide statistical methods for analyzing protein microarray data. It uses a publicly available protein array dataset and emphasizes practical applications in statistics using R, a statistical software. A wide range of statistical methods will be demonstrated, including descriptive statistics, hypothesis testing, false discovery rate, receiver operating characteristic curve, correlation, visualization, and power analysis. The R code used to perform the statistical analyses will be provided.The eye is a multifaceted organ organized in several compartments with particular properties that reflect their diverse functions. The prevalence of ocular diseases is increasing, mainly because of its relationship with aging and of generalized lifestyle changes. However, the pathogenic molecular mechanisms of many common eye pathologies remain poorly understood. Considering the unquestionable importance of proteins in cellular processes and disease progression, proteomic techniques, such as protein microarrays, represent a valuable approach to analyze pathophysiological protein changes in the ocular environment. This technology enables to perform multiplex high-throughput protein expression profiling with minimal sample volume requirements broadening our knowledge of ocular proteome network in eye diseases.In this review, we present a brief summary of the main types of protein microarrays (antibody microarrays, reverse-phase protein microarrays, and protein microarrays) and their application for protein change detection in chronic ocular diseases such as dry eye, age-related macular degeneration, diabetic retinopathy, and glaucoma. The validation of these specific protein changes in eye pathologies may lead to the identification of new biomarkers, depiction of ocular disease pathways, and assistance in the diagnosis, prognosis, and development of new therapeutic options for eye pathologies.The completion and annotation of the human proteome require the availability of information related to protein function. Currently, more than 1800 human genes constitute the "dark proteome," which include missing proteins, uncharacterized human genes validated at protein level, smORFs, proteins from lncRNAs, or any uncharacterized transcripts. During the last years, different experimental workflows based on multi-omics analyses, bioinformatics, and in vitro and in vivo studies have been promoted by the Human Proteome Project Consortium to enhance the annotation of dark proteins. In this chapter, we describe a method that utilizes recombinant proteins and antibody arrays to establish a straightforward methodology in order to rapidly characterize potential functional features of dark proteins associated to intracellular signaling dynamics and extracellular immune response in human cell cultures. Further validating the method, this workflow was applied to probe changes in the activation patterns of kinases and transcription factors as well as in cytokine production modulated by the dark C1orf128 (PITHD1) protein in human olfactory neuroepithelial cells.The heterogeneity of diseases such as cancer makes it necessary to use high-throughput screening techniques to obtain the maximum number of parameters and characteristics of tumors. These obtained biomarkers can be used for the prediction, prognosis, and treatment or search for new therapeutic targets. In this sense, microarray technology allows exhaustive analysis in a short time and from a great variety of biological samples, becoming a fundamental tool in biomedical research projects. Here, operational process of protein microarrays based on the antibody-antigen interaction is described, emphasizing their application in intracellular signaling pathways in tumoral pathologies. In addition, a final validation using nucleic acid programmable protein array (NAPPA) technology in a simple ELISA assay was included to decipher functional characterization of featured proteins from microarray screening.Protein microarrays are a diverse and high-throughput platform for screening biomolecular interactions, autoantigens, and protein expression profiles across tissues, etc. Autoantibodies produced against aberrant protein expression are often observed in malignancies which makes protein microarrays a powerful platform to elucidate biomarkers of translational interest. Early diagnosis of malignancies is an enduring clinical problem that has a direct impact on disease prognosis. Here, we provide an overview of a method employed to screen autoantibodies using patient sera in brain tumors. In case of brain malignancies, early diagnosis is particularly challenging and often requires highly invasive brain biopsies as a confirmatory test. This chapter summarizes the various considerations for applying a serum-based autoantibody biomarker discovery pipeline that could provide a minimally invasive initial diagnostic screen, potentiating classical diagnostic approaches.Since a decade, the nucleic acid protein programmable array (NAPPA) technology has provided researchers with a high-throughput proteomic technique for deciphering immune signatures and screening for biomarkers, among other applications. In osteoarthritis (OA), the activation of proinflammatory pathways of innate immunity due to cellular stress response and cartilage degradation is now considered to be one of the pathophysiological drivers of the disease, perpetuating the catabolic process and the inflammation of the joint. see more Therefore, the identification of immunosignatures in OA would allow to detect autoantibodies (AAbs) as a new source of disease-specific biomarkers. Herein, we describe a proteomic strategy combining NAPPA and in situ protein expression coupled with enzyme-linked immunoassays (ELISA) to search for novel serum AAbs in OA.
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