Search results for "Assay"
showing 10 items of 2241 documents
Evaluation of the Cytotoxicity of Cholesterol Oxides in Human Colon Cancer Caco-2 Cells
2014
The content of cholesterol oxides (COPs) in foods varies between 0.1 and 294.3 mg/g. These oxides are formed by auto-oxidative enzymatic processes promoted by the heat treatment of food and/or to exposition of them to the presence of oxygen and sunlight during storage. Their importance is that they are associated with pathological processes like apoptosis, dyslipidemia and pro-oxidative states, among others. The objective was to evaluate the cytotoxicity, by means MTT assay, of 7-keto cholesterol (7KC), cholestane-triol (Triol), α-epoxy cholesterol (α-epoxy C), β-epoxy cholesterol (β-epoxy C) in differentiated Caco-2 cells at 120μM for 24, 48 and 72h.Triol was the most cytotoxic COP, with a…
Protein Quantitation: Bradford Method
2002
Protein quantitation according to the Bradford method. Keywords: protein quantitation; bradford; coomassie brilliant blue; calorimetric quantification
The Chorioallantoic Membrane Assay in Nanotoxicological Research—An Alternative for In Vivo Experimentation
2020
Nanomaterials unveil many applicational possibilities for technical and medical purposes, which range from imaging techniques to the use as drug carriers. Prior to any human application, analysis of undesired effects and characterization of their toxicological profile is mandatory. To address this topic, animal models, and rodent models in particular, are most frequently used. However, as the reproducibility and transferability to the human organism of animal experimental data is increasingly questioned and the awareness of animal welfare in society increases at the same time, methodological alternatives are urgently required. The chorioallantoic membrane (CAM) assay is an increasingly popu…
All-trans to 11-cis retinol isomerization in nuclear membrane fraction from bovine retinal pigment epithelium
1991
Abstract Isomerization of all-trans to 11-cis retinol has been studied in a membrane preparation from the nuclear fraction of bovine retinal pigment epithelium. When the nuclear membrane preparation deprived of endogenous retinoids is incubated with 4·5 μ m all-trans-retinol, the mean value calculated for the isomerase activity is 1·32 nmol 11-cis retinol formed hr−1 mg protein−1. Simultaneous formation of all-trans and 11-cis retinyl esters is also observed in the nuclear preparation. When assayed under the same experimental condition, RPE 150 000 g post-nuclear sediment shows about 70% of the isomerase activity found in the nuclear membrane fraction. Treatment of the nuclear membrane frac…
Bisfenola A saturošo zobārstniecības kompozītu genotoksicitāte
2019
Daži zobārstniecībā izmantojamie sveķu kompozīti satur monomērus ar BPA kā bāzes elementu, kas izraisa dzīvo šūnu genotoksicitāti un spēj izraisīt dzīvo šūnu nāvi. Šie monomēri spēj noārdīties cilvēka siekalās esošo esterāžu iedarbībā, tādējādi izdalot toksiskos noārdīšanās produktus mutes dobumā. Izdalītie monomēri pat var iekļūt asinsritē, izejot cauri zoba struktūrām uz pulpu. Šī pētījuma mērķis bija izpētīt zobārstniecības kompozīta materiālu ietekmi uz cilvēka asins šūnām, tieši uz limfocītiem, un novērtēt šūnu DNS bojājumu līmeni. Lai izpētītu šo jautājumu, 4 dažādi komerciāli pieejamie zobārstniecības kompozīta materiālu fragmenti tika izmērcēti izolētās cilvēka perifēro asiņu mononu…
Heparan sulfate levels in mucopolysaccharidoses and mucolipidoses.
2004
Glycosaminoglycans are accumulated in both mucopolysaccharidoses (MPS) and mucolipidoses (ML). MPS I, II, III and VII and ML II and ML III patients cannot properly degrade heparan sulphate (HS). In spite of the importance of HS storage in the metabolic pathway in these diseases, blood and urine HS levels have not been determined systematically using a simple and economical method. Using a new ELISA method using anti-HS antibodies, HS concentrations in blood and urine were determined in MPS and ML II and ML III patients. HS concentrations were determined in 156 plasma samples from MPS I (n = 23), MPS II (n = 26), MPS III (n = 24), MPS IV (n = 62), MPS VI (n = 5), MPS VII (n = 5), ML II (n = …
An efficient Escherichia coli expression system for the production of a functional N-terminal domain of the T1R3 taste receptor.
2012
http://www.landesbioscience.com/; International audience; Sweet taste is mediated by a dimeric receptor composed of two distinct subunits, T1R2 and T1R3, whereas the T1R1/T1R3 receptor is involved in umami taste perception. The T1R1, T1R2, and T1R3 subunits are members of the small family of class C G protein-coupled receptors (GPCRs). The members of this family are characterized by a large N-terminal domain (NTD), which is structurally similar to bacterial periplasmic-binding proteins and contains the primary ligand-binding site. In a recent study, we described a strategy to produce a functional dimeric human T1R3-NTD. Although the protein was expressed as inclusion bodies (IBs) using the …
WRN protects against topo I but not topo II inhibitors by preventing DNA break formation
2008
The Werner syndrome helicase/3′-exonuclease (WRN) is a major component of the DNA repair and replication machinery. To analyze whether WRN is involved in the repair of topoisomerase-induced DNA damage we utilized U2-OS cells, in which WRN is stably down-regulated (wrn-kd), and the corresponding wild-type cells (wrn-wt). We show that cells not expressing WRN are hypersensitive to the toxic effect of the topoisomerase I inhibitor topotecan, but not to the topoisomerase II inhibitor etoposide. This was shown by mass survival assays, colony formation and induction of apoptosis. Upon topotecan treatment WRN deficient cells showed enhanced DNA replication inhibition and S-phase arrest, whereas af…
Comparative Surface Morphology, Chemical Composition, and Cytocompatibility of Bio-C Repair, Biodentine, and ProRoot MTA on hDPCs
2020
Biocompatibility is an essential property for any vital pulp material that may interact with the dental pulp tissues. Accordingly, this study aimed to compare the chemical composition and ultrastructural morphology of Biodentine (Septodont, Saint Maur-des-Fosses, France), ProRoot MTA (Dentsply Tulsa Dental Specialties, Johnson City, TN, USA), and Bio-C Repair (Angelus, Londrina, PR, Brazil), as well as their biological effects on human dental pulp cells. Chemical element characterization of the materials was undertaken using scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX). The cytotoxicity was assessed by analyzing the cell viability (MTT assay), cell morphology …
Functional stability of the nitrate-reducing community in grassland soils towards high nitrate supply
2006
Abstract To study the effects of short-term fluctuation of nitrate concentrations on the nitrate-reducing community, repacked soil cores were amended with 0, 100 and 300 μ g NO 3 - ‐ N g - 1 soil and incubated for 3, 7 and 14 days, respectively. The nitrate reductase activity was determined in a laboratory-based enzyme assay. In parallel, the community structure of nitrate-reducing microorganisms was characterised by RFLP-PCR using the functional gene narG , which encodes the catalytic site of the membrane-bound nitrate reductase. The community structure remained constant over the experimental period indicating that this functional community is characterised by a high resistance towards flu…