In-situ hybridization of nodulin mRNAs in root nodules using non-radioactive probes
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In recent years, the development of in situ technologies has made good progress. In situ hybridization (ISH) has become an important tool and has enabled the pathologist to demonstrate infectious pathogens or mRNAs in tissue sections or cytospins without destruction of morphology, thus enabling the assignment of signals to individual cells or cell compartments (, , , , , , , , ).
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Improved methods for both in situ hybridization and in situ enzyme histochemistry were described. The procedures for both methods have been significantly simplified by omitting some unnecessary treatments and substituting the cumbersome and laborious techniques, and the reliability of in situ histochemistry was increased by reversing the operations of Block and Debrouwer's procedure. The improved steps are: Instead of the conventional fixation, a simplified FAA procedure by adding liquid nitrogen onto the embedded tissues during sectioning to ensure high quality of the sections;labeled DNA by randompriming, other than labeled RNA by transcription, was used as probes in hybridization, which was conducted in a moisturesaturated plastic chamber other than emerging in mineral oil. The improved procedure for tissue in situ histochemical study was that the GUS coloration was carried out before fixation, embedding and sectioning, which was different from the procedure as described by Block and Debrouwer.
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The field of metabolomics for cancer diagnosis and characterization is relatively new, and especially the application of metabolomics for in-vivo cancer imaging is still in its infancy. Cancer metabolomics involves the study of global variations of metabolites, with which malignancy conditions can be evaluated by profiling the entire measurable metabolome, instead of focusing only on certain metabolites or isolated metabolic pathways. At present, the study of cancer metabolomics is mainly accomplished utilizing magnetic resonance spectroscopy (MRS) and mass spectrometry (MS). These studies aim to uncover disease-specific metabolomic profiles in order to better understand variations in the involved pathways of cancer metabolism, and to utilize these profiles to establish metabolomic criteria for cancer detection, characterization, and patient prognostication and monitoring. The ultimate goal for cancer metabolomics is its implementation in the clinic to improve clinical abilities for cancer diagnosis and characterization, and through the development of in-vivo metabolomic imaging these clinical goals can be achieved noninvasively.
Keywords:
cancer;
metabolomics;
metabolomic imaging;
magnetic resonance spectroscopy;
mass spectrometry
Metabolome
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Hybridization probe
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Metabolomics, the analysis of the metabolite profile in body fluids or tissues, is being applied to the analysis of a number of different diseases as well as being used in following responses to therapy. While genomics involves the study of gene expression and proteomics the expression of proteins, metabolomics investigates the consequences of the activity of these genes and proteins. There is good reason to think that metabolomics will find particular utility in the investigation of inflammation, given the multi-layered responses to infection and damage that are seen. This may be particularly relevant to eye disease, which may have tissue specific and systemic components. Metabolomic analysis can inform us about ocular or other body fluids and can therefore provide new information on pathways and processes involved in these responses. In this review, we explore the metabolic consequences of disease, in particular ocular conditions, and why the data may be usefully and uniquely assessed using the multiplexed analysis inherent in the metabolomic approach.
Functional Genomics
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Each cell contains many different metabolites and chemical molecules which are generated during cellular process. All the metabolites present in a cell at a particular time is called metabolome. The study of all the metabolites and their modification in a particular condition is called metabolomics. Metabolome is closely linked with genotype, physiology and environment. So,in a nutshell, metabolomics is the study of substrates and products of metabolism which are influenced by the genetic and environmental factors. In plants, metabolomics has now been frequently developed and studied in biotic and abiotic stress resistance. High throughput metabolomics includes time efficient and effective metabolite profiling techniques. These techniques are chromatography based and chromatography free methods. Chromatographic methods are NMR, GC-MS and LC-MS . Chromatographv free techniques include DI-MS,FI-MS,MALDI and Ambient MS . This paper will give an idea about how metabolomics work in elucidating plants phenotype, how sample is prepared for metabolite profiling, different techniques of metabolite profiling and various metabolomic databases.
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Metabolite profiling
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Metabolomics is a field of systems biology that draws on the scientific methods of other groups to qualitatively or quantitatively characterize small molecule metabolites in organisms, revealing their interconnections with the state of the organism at an overall relative macroscopic level. Diabetic kidney disease (DKD) is well known as a chronic metabolic disease, and metabolomics provides an excellent platform for its clinical study. A growing number of metabolomic analyses have revealed that individuals with DKD have metabolic disturbances of multiple substances in their bodies. With the continuous development and improvement of metabolomic analysis technology, the application of metabolomics in the clinical research of DKD is also expanding. This review discusses the recent progress of metabolomics in the early diagnosis, disease prognosis, and pathogenesis of DKD at the level of small molecule metabolites in vivo.
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Abstract Hyperuricemia (HUA) seriously harms human health but the exact etiology and pathogenesis of HUA are not fully understood. Therefore, it is still of great significance to find effective biomarkers and explore the pathogenesis of HUA. Metabolomics reflects the influence of internal and external factors on system metabolism, explains the changes in metabolite levels during the development of diseases, and reveals the molecular mechanism of pathogenesis. Metabolomics is divided into untargeted metabolomics and targeted metabolomics according to different research modes. Each other's advantages can be fully utilized by combining the two so that the results of metabolomics research can be consummated. 20 HUA patients and 20 healthy individuals participated in the experiment, and untargeted metabolomics was employed to find 50 differential metabolites in HUA serum samples. Twelve candidate biomarkers were screened based on literature research and ROC Curve analysis for subsequent verification. Based on the UPLC-TQ-MS analysis platform, the targeted metabolomics detection methods were established and the content of 12 candidate biomarkers was precisely quantified. Compare with the results of untargeted metabolomics, the targeted metabolomics results were considered more reliable.
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