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Fluorescent Technology in the Assessment of Metabolic Disorders in Diabetes
Published in Andrey V. Dunaev, Valery V. Tuchin, Biomedical Photonics for Diabetes Research, 2023
Elena V. Zharkikh, Viktor V. Dremin, Andrey V. Dunaev
This chapter reviews and analyzes the prospects for the use of fluorescence techniques in the diagnosis and assessment of the severity of diabetic complications. Methods of fluorescence spectroscopy and imaging are considered to be promising modern methods of optical diagnostics. Noninvasive assessment of fluorescence of biological tissues allows us to draw conclusions about structural and metabolic changes that occur in various pathological processes. Fluorescence spectroscopy and imaging are widely used in various medical fields, including oncology and skin diagnostics.
Basic genetics and patterns of inheritance
Published in Hung N. Winn, Frank A. Chervenak, Roberto Romero, Clinical Maternal-Fetal Medicine Online, 2021
Fluorescence in situ hybridization (FISH) is a technique in which DNA probes attached to fluorescent dyes are hybridized to specific chromosome regions from patient samples, either interphase cells or metaphase chromosome spreads, most often from lymphocytes or amniocytes. The chromosomes are then visualized by fluorescent microscopy. Probes can be designed to hybridize to whole chromosomes, specific chromosome segments of interest, centromeres, or telomeres. FISH is widely used for the diagnosis of suspected recognizable microdeletion syndromes (Fig. 4). Multicolor FISH is useful for analyzing submicroscopic structural rearrangements undetectable by classic cytogenetic techniques or for identifying marker chromosomes. FISH is also used for rapid screening for aneuploidy including trisomy 13, 18, or 21 and abnormalities of X and Y.
Conjugation and Other Methods in Polymeric Vaccines
Published in Mesut Karahan, Synthetic Peptide Vaccine Models, 2021
These two processes can explain these; determination of fluorescence life (lifetime), investigation of properties of excised molecules, definition of complex formation mechanism, investigation of solvent-soluble interaction, isomerization, diffusion, investigation of micellar properties, surface properties, polymer structure and dynamics, enzyme structure, investigation of antigen-antibody interaction, biomembrane. The use of the fluorescence resonance energy transfer (FRET) method to explain the relationship between the donor and the acceptor is an example of the use of fluorescence spectrometers. Furthermore, this method is very important in amino acid detection. Tryptophan (Trp, W), tyrosine (Tyr, Y), and phenylalanine (Phe, F) are fluorescent forms of amino acids present in the structure of proteins and peptides. Among these tryptophan is the amino acid with the highest fluorescence (Ghisaidoobe and Chung 2014).
MicroRNA-122-5p ameliorates tubular injury in diabetic nephropathy via FIH-1/HIF-1α pathway
Published in Renal Failure, 2022
Li Cheng, Xinying Qiu, Liyu He, Li Liu
Fluorescence in situ hybridization (FISH) was performed according to the manufacturer's instructions. Briefly, kidneys were harvested from control and STZ-treated mice to prepare 4-micron paraffin section. The sections were treated with 20 μg/ml proteinase K for permeabilization, and then incubated with pre-hybridization solution at 78 °C for 1 h. Remove pre-hybridization solution and add digoxigenin-labeled mmu-miR-122-5p LNA probe over night at 37 °C. At the second day, after wash, bovine serum albumin (BSA) was added for blocking. Then the anti-digoxigenin-HRP was used at 37 °C for 1 h. CY3-TSA and DAPI assay were used to indicate the positive areas and cell nucleus respectively. The images were acquired from a fluorescence microscope and the representative figures were exhibited.
Immunologic evaluation and genetic defects of apoptosis in patients with autoimmune lymphoproliferative syndrome (ALPS)
Published in Critical Reviews in Clinical Laboratory Sciences, 2021
Laura Casamayor-Polo, Marta López-Nevado, Estela Paz-Artal, Alberto Anel, Frederic Rieux-Laucat, Luis M. Allende
The molecular study of somatic mutations in the FAS gene is mandatory in patients with elevated DNTs and biomarkers and in whom a germline mutation has not been identified. The workflow for genetic testing of somatic mutations (ALPS-sFAS) differs from that for germline mutations (Figure 6). Instead of working with DNA extracted from a whole blood sample, it is necessary to extract DNA from DNTs. For this application, the amount of whole blood required is higher (approximately 15 mL) because of the need to isolate DNTs using magnetic-activated cell sorting separation (Double-negative T Cell Isolation Kit (Miltenyi Biotec GmbH, Germany). Reference: 130-092-614) or fluorescence-activated cell sorting (FACS). For magnetic-activated cell sorting, the isolation of DNTs is performed in a two-step procedure: firstly, the CD4 + CD8+ and CD56+ cells are depleted from peripheral blood mononuclear cells (PBMC) and secondly, a positive selection of TCRαβ + CD4-CD8- cells are retained. For FACS sorting, PBMCs are stained with the same monoclonal antibody cocktail as for DNT immunophenotyping. In both cases, cell purity analysis is essential to ensure that there are more than 90% of DNTs. Finally, it is important to mention that somatic mutations in the FAS gene are restricted mainly to exons 7, 8, and 9 (Figure 6(A)).
Clinical role of fluorescence imaging in colorectal surgery - an updated review
Published in Expert Review of Medical Devices, 2020
Amandeep Ghuman, Sandra Kavalukas, Stephen P. Sharp, Steven D. Wexner
The role of fluorescence imaging has been expanding. It was initially used to assess bowel perfusion to ensure well-vascularized anastomoses were created with the hypothesis this objective assessment may lead to reduction in AL. Although the results of the two RCTs are conflicting and the preliminary data from PILLAR III did not establish a clear correlation, likely due to small sample size since study was terminated early, the potential benefits may still exist. Establishing a clear role for ICG in AL reduction will require consistency in the definition of AL used in the literature, along with standardized anastomotic measurements, as more distal (lower) anastomoses are known to carry higher risk of AL than are the more proximal (higher) ones. In the current literature there does appear to be a reduction in AL with ICG for rectal cancer resections, which could be due to higher AL at baseline with these lower anastomoses. Given the multifactorial nature of risk factors for colorectal AL, larger multi-center RCTs may be required to delineate a clear role.