Cytochrome P450 (P450) and uridine 5′-diphospho-glucuronosyltransferase (UGT) enzymes mediate a major proportion of phase I II metabolism xenobiotics. In vitro-in vivo extrapolation (IVIVE) hepatic clearance in conjunction with physiologically-based pharmacokinetics (PBPK) has become common practice drug development. However, prediction xenobiotic kinetics virtual populations requires knowledge both enzyme abundances the extent to which these correlate. A multiplexed quantification...

The blood-brain barrier (BBB) maintains brain homeostasis by controlling traffic of molecules from the circulation into brain. This function is predominantly dependent on proteins expressed at BBB, especially transporters and tight junction proteins. Alterations to level BBB can impact susceptibility central nervous system exposure xenobiotics in systemic with potential consequent effects function. In this study, expression profiles drug solute carriers were assessed tissues healthy...

Glutathione reductase (Glr1) is a low abundance protein involved in defense mechanisms against reactive oxygen species. Expressed on cytosolic ribosomes, the same gene, GLR1, uses alternative start codons to generate two forms of Glr1. Translation from first AUG codon generates mitochondrial form incorporating presequence necessary for import; translation second yields counterpart. Proteomic strategies were used analyze N-terminal sequences and turnover Saccharomyces cerevisiae The...

Quantitative translation of information on drug absorption, disposition, receptor engagement, and drug–drug interactions from bench to bedside requires models informed by physiological parameters that link in vitro studies vivo outcomes. To predict outcomes, biochemical data experimental systems are routinely scaled using protein quantity these relevant tissues. Although several laboratories have generated useful quantitative proteomic state‐of‐the‐art mass spectrometry, no harmonized...

The levels of drug-metabolizing enzymes (DMEs) and transporter proteins in the human intestine are pertinent to determine oral drug bioavailability. Despite paucity reports on such measurements, it is well recognized that these values essential for translating vitro data metabolism transport predict disposition gut wall. In current study, clinically relevant DMEs [cytochrome P450 (P450) uridine 5′-diphospho-glucuronosyltransferase (UGT)] transporters were quantified total mucosal protein...

Twenty‐seven cases of neosporosis in European dogs are described. The disease was confirmed by immunohistochemistry, electron microscopy, or a favourable response to treatment the with appropriate clinical signs, and presence antibodies Neospora caninum but not Toxoplasma gondii . affected were two days seven years old, 13 different breeds. Both sexes most littermates remained normal. Twenty‐one had an initial hindlimb paresis ataxia, which muscle atrophy consistent sign. Rigid...

A total of 1,554 dogs from 5 countries on 3 continents were tested for antibodies to Neospora caninum using an indirect fluorescent antibody test. In Australia, overall, 42/451 (9%, 95% confidence interval [CI] 6-12%) seropositive (Melbourne 11/207 [5%, CI 2-9%]; Sydney 18/150 [12%, 7-18%]; Perth 13/94 [14%, 8-22%]). Antibodies N. also detected in South America (Uruguay [20%, 16-24%, n = 414]) and sub-Saharan Africa (Tanzania [22%, 12-36%, 49]). contrast, only 1 500 the Falkland Islands none...

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTMinor-groove recognition of the self-complementary duplex d(CGCGAATTCGCG)2 by Hoechst 33258: a high-field NMR studyJohn A. Parkinson, Jill Barber, Kenneth T. Douglas, John Rosamond, and Derek SharplesCite this: Biochemistry 1990, 29, 44, 10181–10190Publication Date (Print):November 1, 1990Publication History Published online1 May 2002Published inissue 1 November...

The cytochrome P450 (P450) family of enzymes is a major player in the metabolism therapeutic drugs available on market, and development novel has to take into account these fate new drugs. Testing pharmacokinetic behavior animals common part drug process. Pigs are increasingly used for this purpose because their similarity enzymatic pattern humans. In study, adult Suffolk White pig liver microsomal samples were analyzed using mass-spectrometry-based techniques identify relatively quantify...

In the present work, two different proteomic platforms, gel-based and gel-free, were used to map matrix outer membrane vesicle exoproteomes of Pseudomonas aeruginosa PAO1 biofilms. These strategies allowed us a confident identification 207 327 proteins from enriched vesicles whole isolated Because physicochemical characteristics these subproteomes, showed complementarity, thus, most comprehensive analysis P. exoproteome date was achieved. Under our conditions, contribute approximately 20%...

There is an urgent need (recognized in FDA guidance, 2018) to optimize the dose of medicines given patients for maximal drug efficacy and limited toxicity (precision dosing), which can be facilitated by quantitative systems pharmacology (QSP) models. Accurate quantification proteins involved clearance essential build improve QSP models any target population. Here we describe application label-free proteomics microsomes from 23 human livers simultaneously quantify 188 enzymes 66 transporters...

Variability in individual capacity for hepatic elimination of therapeutic drugs is well recognized and associated with variable expression activity liver enzymes transporters. Although genotyping offers some degree stratification, there often large variability within the same genotype. Direct measurement protein impractical due to limited access tissue biopsies. Hence, determination drug metabolism disposition using liquid biopsy (blood samples) an attractive proposition during development...

Abstract The blood–brain barrier ( BBB ) maintains brain homeostasis by tightly regulating the exchange of molecules with systemic circulation. It consists primarily microvascular endothelial cells surrounded astrocytic endfeet, pericytes, and microglia. Understanding make‐up transporters in rat is essential to translation pharmacological toxicological observations into humans. In this study, experimental workflows are presented which optimization (a) isolation microvessels (b) enrichment...

Liver cirrhosis is a chronic disease that affects the liver structure, protein expression, and overall metabolic function. Abundance data for drug-metabolizing enzymes transporters (DMET) across all stages of severity are scarce. Levels these proteins crucial accurate prediction drug clearance in hepatically impaired patients using physiologically based pharmacokinetic (PBPK) models, which can be used to guide selection more precise dosing. This study aimed experimentally quantify human...

SUMMARY An indirect enzyme linked immunosorbent assay (ELISA) for detection of antibodies to Neospora caninum in serum from dogs is described. Extracted tachyzoite proteins incorporated into immunostimulating complexes (iscoms) were used as coating antigen. A mixture a monoclonal antibody dog immunoglobulin G and horse radish peroxidase conjugated mouse Ig was detect bound antibody. When the iscom preparation analysed by means sodium dodecyl sulphate polyacrylamide gel electrophoresis it...

SUMMARY Neospora caninum is an apicomplexan, protozoan parasite, which causes severe disease in dogs and cattle. It has previously been isolated only the United States. A 5-week-old Boxer pup with a progressive hindlimb paresis was diagnosed as suffering from neosporosis on basis of clinical signs presence anti- antibodies it, 2 litter-mates its darn. Despite treatment sulphonamides, euthanased 3 days later. The diagnosis confirmed by immunohistochemical examination muscle CNS tissue...

QconCAT is a tool for quantitative proteomics, consisting of an artificial protein, expressed from gene, made up concatenated string proteotypic peptides selected the proteins under study. Isotopically labeled (usually containing 13C6-arginine and 13C6-lysine) provides standard each peptide included in its sequence. In practice, some fail to express at sufficient levels purpose analysis. Two complementary methods are presented recalcitrant intended quantify human hepatic enzymes transporters.

Many genetic and environmental factors lead to interindividual variations in the metabolism transport of drugs, profoundly affecting efficacy toxicity. Precision dosing, that is, targeting drug dose a well characterized subpopulation, is dependent on quantitative models profiles drug-metabolizing enzymes (DMEs) transporters within informed by proteomics. We report first use ion mobility–mass spectrometry for this purpose, allowing rapid, robust, label-free quantification human liver...

Quantitative characterization of UDP-glucuronosyltransferase (UGT) enzymes is valuable in glucuronidation reaction phenotyping, predicting metabolic clearance and drug-drug interactions using extrapolation exercises based on pharmacokinetic modeling. Different quantitative proteomic workflows have been employed to quantify UGT various systems, with reports indicating large variability expression, which cannot be explained by interindividual alone. To evaluate the effect methodological...