A Review on Ethnobotany of Hepatoprotective Plants of India
T. Pullaiah, K. V. Krishnamurthy, Bir Bahadur in Ethnobotany of India, 2017
The liver biopsy results may not be comparable with the LFTS since many functional changes are not mirrored by obvious structural changes in the liver cells (Praful and Drashan, 1996). Thus a battery of liver function tests is employed for accurate diagnosis, to assess the severity of the damage, to judge the prognosis and to evaluate therapy. These tests are described below in relation to major liver functions.Tests for manufacture and excretion of bile (Mohan, 2005).Serum enzyme assay include Alkaline phosphatase, Transaminases: SGOT (AST) and SGPT (ALT), γ-Glutamyl transpeptidase (γ-GT), 5-Nucleotidase and lactic dehydrogenase.Immunologic tests.
A Rapid Purification of Dna Topoisomerase I, A Chromatin-Bound Nonhistone Protein, and its Inhibition by Heparin
Isaac Bekhor, Carol J. Mirell, C. C. Liew in Progress in Nonhistone Protein Research, 1985
Cultured mouse mammary carcinoma cell line FM3A was used as the source of DNA topoisomerase I. Chromatin extract containing all the enzyme activity of the cells was prepared by the method of Germond et al.25 and used as a starting material for the purification of the enzyme. A unit of enzyme is defined as the activity that converts 1 μg of col El DNA I to fully relaxed form Ir after incubation at 37°C for 15 min. Further details of enzyme assay were described in a previous article.24 SDS-PAGE (polyacrylamide gel electrophoresis) of proteins was performed by the prescription of Laemmli26 and Blattler et al.27 Recovery of the enzyme from SDS-polyacrylamide gel was performed according to the method of Hager and Burgess.28 Heparin-Sepharose® was prepared as described.29 Phenyl-Sepharose® was purchased from Pharmacia Fine Chemicals, Uppsala, Sweden. Hog intestinal heparin was obtained from Cohelfred Laboratories, Chicago, and purified free of dermatan sulfate by the method of Rodén et al.30 Affinity-chromatography of heparin on antithrombin III-Sepharose® was performed essentially as described by Laurent et al.31 Gel-filtration chromatography of heparin samples was performed on Ultrogel® AcA44 agarose-acrylamide gel obtained from LKB-Produkter AB, Broma, Sweden. Heparin samples were electrophoresed on cellulose acetate membrane by the methods of Seno et al.32 and Wessler.33 Anticoagulant activity of heparin was assayed by the whole-blood assay method of the USP,34 and the activity was expressed as units per milligram. Chemical analyses of heparin samples were performed as described.35
Ria: Principles and Application
Fuad S. Ashkar, Lelio G. Colombetti in Radiobioassays, 2019
The heterogeneous type of enzyme assay has several variations. The competitive antigen enzyme linked immunosorbent assay (ELISA) involves an antigen covalently linked to a solid-phase (a tube or beads), the addition of enzyme-labeled antibody and the test antigen (analyte), subsequent separation of either fraction, addition of substrate, and finally quantitation.92 Another type of immunosorbent assay using a double antibody technique or “sandwich” technique has found application in detection of hepatitis, rubella, and toxoplasma as well as other pathogenic organisms.93, 94
An experimental investigation into cardiovascular, haemodynamic and salivary alpha amylase reactivity to acute stress in Type D individuals
Published in Stress, 2019
Sarah F. Allen, Mark A. Wetherell, Michael A. Smith
The passive drool technique was implemented to collect saliva. Participants were instructed to allow saliva to pool in the bottom of their mouth, and then pass the saliva through a SalivaBio Collection Aid (SCA) into a 4 ml polypropylene tube for 2 min. Samples were taken at baseline, pre-stressor, and post-stressor, in addition to +10 min and +20 min post-stressor. As alpha amylase has a diurnal profile (O’Donnell, Kammerer, O’Reilly, Taylor, & Glover, 2009), testing occurred at 2 pm each day. Samples were immediately stored at −20 C and transferred to −80 C within 24 h. On day of assay, samples were completely thawed and centrifuged at 1500 × g for 15 min. Assays were conducted in-house by a trained technician using a kinetic enzyme assay kit provided by Salimetrics. Saliva flow rate (mL/min) was calculated and alpha amylase output (U/min) computed for each sample. All samples were assayed in duplicate. Intra-assay variation (CV) was computed for the mean of duplicate samples and those with a CV above 15% excluded from analyses. This resulted in 56 participants providing full alpha amylase data (31 non-Type D, 25 Type D). Inter-assay variation was below 15% and therefore deemed acceptable.
Recent advances in molecular testing to improve early diagnosis in children with mucopolysaccharidoses
Published in Expert Review of Molecular Diagnostics, 2018
Ana Carolina Brusius-Facchin, Diana Rojas Malaga, Sandra Leistner-Segal, Roberto Giugliani
The diagnosis of MPS is based mainly in biochemical and molecular testing. Usually the first step in the diagnosis pathway is the urine screening with quantitation and qualitative evaluation (by electrophoresis or thin-layer chromatography) of urinary GAGs [55,56]. These results, together with the clinical findings, can help guiding the choice of the confirmatory enzyme assay to be performed. Assays to measure activity of specific enzymes in leukocytes or fibroblasts are considered the gold standard for a definitive diagnosis. For some enzymes, assays in plasma are also available. In some cases, it is possible to perform the enzyme assay in dried blood spots (DBS) [10]. This option is instrumental in regions where collecting and shipping whole blood or other tissue samples is impractical. The present recommendation is that positive results in DBS should be confirmed in leucocytes or fibroblasts. When this is not possible, enzyme assays should be performed at least twice in DBS (in two independent samples) and/or should have the results confirmed by genotyping [10]. Although the finding of a specific enzyme deficiency in leucocytes or fibroblasts confirms the diagnosis, molecular analysis of the respective gene is recommended, whenever possible (Figure 1).
Kushenol A and 8-prenylkaempferol, tyrosinase inhibitors, derived from Sophora flavescens
Published in Journal of Enzyme Inhibition and Medicinal Chemistry, 2018
Jang Hoon Kim, In Sook Cho, Yang Kang So, Hyeong-Hwan Kim, Young Ho Kim
Enzyme assay was performed according to the modified methods in the previous papers7. For the calculation of inhibitory activity, 130μL of tyrosinase (about 46 units/mL) solvated in 0.1mM phosphate buffer (pH: 6.8) and 20μL of 1–0.0078mM concentrations of the inhibitors were mixed in a 96-well plate, and then 50μL of 2mM L-tyrosine in buffer was added in mixture. To test the enzyme kinetic study, 130μL of tyrosinase and 20μL of inhibitor were also mixed, and then 50μL of 0.62–10mM L-tyrosine was added in a 96-well plate. The mixture was recorded at UV-Vis 475nm during 20 min. The inhibitory ratio was calculated according to the following equation: C20 and S20 are the intensity of control and inhibitor after 20min, C0 and S0 are the intensity of control and inhibitor at 0 min.
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