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Antioxidants (Basel, Switzerland) Mar 2022Common peroxidase action and haloperoxidase action are quantifiable as light emission from dioxygenation of luminol (5-amino-2,3-dihydrophthalazine-1,4-dione). The...
Common peroxidase action and haloperoxidase action are quantifiable as light emission from dioxygenation of luminol (5-amino-2,3-dihydrophthalazine-1,4-dione). The velocity of enzyme action is dependent on the concentration of reactants. Thus, the reaction order of each participant reactant in luminol luminescence was determined. Horseradish peroxidase (HRP)-catalyzed luminol luminescence is first order for hydrogen peroxide (HO), but myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are second order for HO. For MPO, reaction is first order for chloride (Cl) or bromide (Br). For EPO, reaction is first order for Br. HRP action has no halide requirement. For MPO and EPO, reaction is first order for luminol, but for HRP, reaction is greater than first order for luminol. Haloperoxidase-catalyzed luminol luminescence requires acidity, but HRP action requires alkalinity. Unlike the radical mechanism of common peroxidase, haloperoxidases (XPO) catalyze non-radical oxidation of halide to hypohalite. That reaction is second order for HO is consistent with the non-enzymatic reaction of hypohalite with a second HO to produce singlet molecular oxygen (O*) for luminol dioxygenation. Alternatively, luminol dehydrogenation by hypohalite followed by reaction with HO yields dioxygenation consistent with the same reaction order. Haloperoxidase action, Cl, and Br are specifically quantifiable as luminol luminescence in an acidic milieu.
PubMed: 35326168
DOI: 10.3390/antiox11030518 -
Discovery Medicine Jul 2019The goal of this study is to evaluate a novel direct immunohistochemistry staining method on frozen tissues for the intraoperative diagnosis of breast papillary lesions.
AIMS
The goal of this study is to evaluate a novel direct immunohistochemistry staining method on frozen tissues for the intraoperative diagnosis of breast papillary lesions.
METHODS AND RESULTS
Keratin 5 (CK5) and smooth muscle myosin heavy chain (SMMHC) antibodies were labeled with horseradish peroxidase polymers and used for direct immunohistochemistry (IHC) staining on frozen sections of breast tissues during surgical operations. The results from direct IHC on 102 cases of breast papillary lesions were compared with those obtained by the conventional staining method on formalin-fixed paraffin-embedded tissues (FFPE). Compared to the conventional method, direct IHC staining can significantly improve the accuracy of intraoperative diagnosis of breast papillary lesions from 70% to 97% (p < 0.01). No false negative cases were found with direct IHC in this study. In comparison, 53% of cases with the conventional method were found false negative. Direct IHC also significantly reduced the deferred diagnosis rate from 21% to 3% (p < 0.01). Furthermore, the entire procedure of direct IHC can be finished within 10 minutes, which makes it more feasible for the use of intraoperative frozen section diagnosis.
CONCLUSION
The direct IHC staining method can significantly improve the efficiency and accuracy of intraoperative diagnosis of breast papillary lesions. It also fits better for the quick turnaround time required for intraoperative diagnosis.
Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Female; Frozen Sections; Humans; Immunohistochemistry; Intraoperative Care; Keratin-5; Middle Aged; Predictive Value of Tests
PubMed: 31465723
DOI: No ID Found -
Sensors (Basel, Switzerland) Feb 2020For the early diagnosis of several diseases, various biomarkers have been discovered and utilized through the measurement of concentrations in body fluids such as blood,... (Review)
Review
For the early diagnosis of several diseases, various biomarkers have been discovered and utilized through the measurement of concentrations in body fluids such as blood, urine, and saliva. The most representative analytical method for biomarker detection is an immunosensor, which exploits the specific antigen-antibody immunoreaction. Among diverse analytical methods, surface plasmon resonance (SPR)-based immunosensors are emerging as a potential detection platform due to high sensitivity, selectivity, and intuitive features. Particularly, SPR-based immunosensors could detect biomarkers without labeling of a specific detection probe, as typical immunosensors such as enzyme-linked immunosorbent assay (ELISA) use enzymes like horseradish peroxidase (HRP). In this review, SPR-based immunosensors utilizing noble metals such as Au and Ag as SPR-inducing factors for the measurement of different types of protein biomarkers, including viruses, microbes, and extracellular vesicles (EV), are briefly introduced.
Topics: Bacteria; Biomarkers; Extracellular Vesicles; Metals; Proteins; Surface Plasmon Resonance
PubMed: 32069896
DOI: 10.3390/s20041003 -
Sensors (Basel, Switzerland) Apr 2020Suitable immobilization of a biorecognition element, such as an antigen or antibody, on a transducer surface is essential for development of sensitive and analytically... (Review)
Review
Suitable immobilization of a biorecognition element, such as an antigen or antibody, on a transducer surface is essential for development of sensitive and analytically reliable immunosensors. In this review, we report on (1) methods of antibody prefunctionalization using electroactive probes, (2) methods for immobilization of such conjugates on the surfaces of electrodes in electrochemical immunosensor construction and (3) the use of antibody-electroactive probe conjugates as bioreceptors and sensor signal generators. We focus on different strategies of antibody functionalization using the redox active probes ferrocene (Fc), anthraquinone (AQ), thionine (Thi), cobalt(III) bipyridine (Co(bpy)), Ru(bpy) and horseradish peroxidase (HRP). In addition, new possibilities for antibody functionalization based on bioconjugation techniques are presented. We discuss strategies of specific, quantitative antigen detection based on (i) a sandwich format and (ii) a direct signal generation scheme. Further, the integration of different nanomaterials in the construction of these immunosensors is presented. Lastly, we report the use of a redox probe strategy in multiplexed analyte detection.
Topics: Antibodies; Antibodies, Immobilized; Antigens; Electrochemical Techniques; Electrodes; Ferrous Compounds; Immunoassay; Metallocenes; Nanostructures; Oxidation-Reduction; Phenothiazines
PubMed: 32260217
DOI: 10.3390/s20072014 -
JACC. Basic To Translational Science Jul 2021Protein-protein interactions are of paramount importance in regulating normal cardiac physiology. Methodologies to elucidate these interactions in vivo have been... (Review)
Review
Protein-protein interactions are of paramount importance in regulating normal cardiac physiology. Methodologies to elucidate these interactions in vivo have been limited. Recently, proximity-dependent biotinylation, with the use of BioID, TurboID, and ascorbate peroxidase, has been developed to uncover cellular neighborhoods and novel protein-protein interactions. These cutting-edge techniques have enabled the identification of subcellular localizations of specific proteins and the neighbors or interacting proteins within these subcellular regions. In contrast to classic methods such as affinity purification and subcellular fractionation, these techniques add covalently bound tags in living cells, such that spatial relationships and interaction networks are not disrupted. Recently, these methodologies have been used to identify novel protein-protein interactions relevant to the cardiovascular system. In this review, we discuss the development and current use of proximity biotin-labeling for cardiovascular research.
PubMed: 34368510
DOI: 10.1016/j.jacbts.2021.01.005 -
Molecules (Basel, Switzerland) Oct 2020Silk fibroin is a widely and commercially available natural protein derived from silkworm cocoons. Thanks to its unique amino acid composition and structure, which lead... (Review)
Review
Silk fibroin is a widely and commercially available natural protein derived from silkworm cocoons. Thanks to its unique amino acid composition and structure, which lead to localized nanoscale pockets with limited but sufficient hydration for protein interaction and stabilization, silk fibroin has been studied in the field of enzyme immobilization. Results of these studies have demonstrated that silk fibroin offers an important platform for covalent and noncovalent immobilization of enzymes through serving as a stabilization matrix/support with high retention of the biological activity of the enzymes of interest. In the hope of providing suggestions for potential future research directions, this review has been written to briefly introduce and summarize key advances in silk fibroin-based materials for immobilization of both enzymes/biocatalysts (including alkaline phosphatase, β-glucosidase, glucose oxidase, lipase, urease, uricase, horseradish peroxidase, catalase, xanthine oxidase, tyrosinase, acetylcholinesterase, neutral protease, α-chymotrypsin, amylase, organophosphorus hydrolase, β-galactosidase, carbonic anhydrase, laccase, zymolyase, phenylalanine ammonia-lyase, thymidine kinase, and several others) and non-enzymatic catalysts (such as Au, Pd, Fe, α-FeO, FeO, TiO, Pt, ZnO, CuO, CuO, MnO, and MnO).
Topics: Animals; Bioprinting; Catalysis; Dendrimers; Enzymes, Immobilized; Fibroins; Humans; Metals; Oxides; Surface Properties
PubMed: 33114465
DOI: 10.3390/molecules25214929 -
Nanomaterials (Basel, Switzerland) Dec 2022In theory, nanoplastics (NPs) can adsorb biological macromolecules, such as proteins, in the surrounding environment to form protein corona (PC). In this study, we focus...
In theory, nanoplastics (NPs) can adsorb biological macromolecules, such as proteins, in the surrounding environment to form protein corona (PC). In this study, we focus on amino polystyrene (PS) NPs and horseradish peroxidase (HRP) to explore the dynamic process of the formation of PS-HRP PC and their influence on PS and HRP. This work used atomic force microscopy, laser particle size and Zeta potential analyzer, and UV-vis spectrophotometer. According to the adsorption behavior of HRP to NPs, the surface morphology characteristics of NPs can be observed to change at 60 min. Meanwhile, the increase in size and hydrodynamic diameter, the decrease in Zeta potential, surface roughness and HRP activity, and the change in HRP structure attest to the PC formation. The thickness of the PC was approximately 30 nm and there are differences in the dynamic and static variations in the size of the PC. The PC formation process progresses gradually from 0 min to 240 min. Overall, the formation of PS-HRP PC is identified, and the changes in its properties are confirmed from the perspective of nanoplastics and peroxidase, which help study the effects of nanoplastics on the environment and creatures.
PubMed: 36558320
DOI: 10.3390/nano12244467 -
Pharmaceuticals (Basel, Switzerland) Oct 2022Dehydrodiisoeugenol (DHIE) is a neolignan found in more than 17 plant species, including herbs, fruit, and root. DHIE was, for the first time, isolated from bark in... (Review)
Review
Dehydrodiisoeugenol (DHIE) is a neolignan found in more than 17 plant species, including herbs, fruit, and root. DHIE was, for the first time, isolated from bark in 1973. Since then, many methodologies have been used for the obtention of DHIE, including classical chemistry synthesis using metal catalysts and biocatalytic synthesis; employing horseradish peroxidase; peroxidase from ; laccase; culture cells of plants; and microorganisms. Increasing evidence has indicated that DHIE has a wide range of biological activities: anti-inflammatory, anti-oxidant, anti-cancerogenic, and anti-microbial properties. However, evidence in vivo and in human beings is still lacking to support the usefulness potential of DHIE as a therapeutic agent. This study's review was created by searching for relevant DHIE material on websites such as Google Scholar, PubMed, SciFinder, Scholar, Science Direct, and others. This reviews the current state of knowledge regarding the different synthetical routes and biological applications of DHIE.
PubMed: 36355523
DOI: 10.3390/ph15111351 -
Bio-protocol Nov 2021CD45 is a pan-leukocyte marker, and CD45 stain is widely used to determine the extent of inflammatory cell infiltration and its association with tissue injury. In this...
CD45 is a pan-leukocyte marker, and CD45 stain is widely used to determine the extent of inflammatory cell infiltration and its association with tissue injury. In this manuscript, we share a reliable immunohistochemistry (IHC) protocol for CD45 staining in sections of paraffin-embedded mouse kidney. A rat anti-CD45 antibody was used as primary antibody, and a mouse adsorbed biotin-conjugated goat anti-rat IgG was selected as secondary antibody. A horseradish peroxidase (HRP)-linked avidin/biotin detection system was used to amplify the signal, which was detected with 3,3'-Diaminobenzidine (DAB). With this protocol, we show that the CD45 antibody recognizes cells of hematolymphoid lineage in bone marrow, as well as monocyte/macrophages in liver and lung tissue. The utility of this protocol in pathology research was indicated by dramatically increased CD45-positive (CD45) cells in the kidneys of a mouse model of diabetes. Double staining for CD45 and injury marker KIM-1 showed accumulated CD45 cells around injured tubular cells. CD45 and F4/80 macrophage staining on adjacent tissue sections revealed overlap of CD45 cells with other inflammatory cells.
PubMed: 34909451
DOI: 10.21769/BioProtoc.4230 -
Sensors (Basel, Switzerland) Jan 2020A convenient electrochemical sensing pathway was investigated for neurotransmitter detection based on newly synthesized silole derivatives and...
A convenient electrochemical sensing pathway was investigated for neurotransmitter detection based on newly synthesized silole derivatives and laccase/horseradish-peroxidase-modified platinum (Pt)/gold (Au) electrodes. The miniature neurotransmitter's biosensors were designed and constructed via the immobilization of laccase in an electroactive layer of the Pt electrode coated with poly(2,6-bis(3,4-ethylenedioxythiophene)-4-methyl-4-octyl-dithienosilole) and laccase for serotonin (5-HT) detection, and a Au electrode modified with the electroconducting polymer poly(2,6-bis(selenophen-2-yl)-4-methyl-4-octyl-dithienosilole), along with horseradish peroxidase (HRP), for dopamine (DA) monitoring. These sensing arrangements utilized the catalytic oxidation of neurotransmitters to reactive quinone derivatives (the oxidation process was provided in the enzymes' presence). Under the optimized conditions, the analytical performance demonstrated a convenient degree of sensitivity: 0.0369 and 0.0256 μA mM cm, selectivity in a broad linear range (0.1-200) × 10 M) with detection limits of ≈48 and ≈73 nM (for the serotonin and dopamine biosensors, respectively). Moreover, the method was successfully applied for neurotransmitter determination in the presence of interfering compounds (ascorbic acid, L-cysteine, and uric acid).
Topics: Biosensing Techniques; Catalysis; Dopamine; Electrochemical Techniques; Electrodes; Enzymes, Immobilized; Gold; Horseradish Peroxidase; Hydrogen-Ion Concentration; Laccase; Limit of Detection; Microscopy, Atomic Force; Neurotransmitter Agents; Oxidation-Reduction; Platinum; Polymers; Serotonin; Silicon Compounds
PubMed: 31940833
DOI: 10.3390/s20020423