Reliability optimisation
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Background: Adverse drug reactions (ADRs), particularly in the context of polypharmacy, remain a persistent, unresolved problem for patients and healthcare professionals. The ADRe Profile identifies medicine-related harms, and supports their resolution, thereby improving care quality and preventing future problems. Objective: The objective of this study was to assess the validity and reliability of the ADRe Profile (https://www.swansea.ac.uk/adre/) in U.K. primary care general practices, building on assessments in other settings. Methods: The ADRe Profile's validity and reliability were investigated using complementary mixed methods: content validity index, contrast group construct validity, cognitive interviewing, and inter-rater reliability. Results: Cognitive interviews (n = 5) confirmed that the ADRe Profile needed only minor adjustments. The scale-level content validity index was 0.67 (n = 14), items ranging from 0.08 to 1. Significant differences in signs and symptoms associated with ADRs between service users taking different numbers of regular prescribed medicines confirmed construct validity (n = 68, U = 870.50, p < 0.001). Inter-rater reliability testing showed substantial agreement between service users and research nurse: 10 items had 100% agreement. Overall kappa mean was 0.71 (range: 0.31-1), (n = 42). Conclusions and Relevance: The ADRe Profile is suitable for use with older service users in primary care who live at home. Users understood the questions and provided meaningful answers. ADRe Profile responses were sufficiently reliable to be used as a basis for further investigations, prescriber referral and clinical actions. However, clinician judgement of content validity may depend on knowledge and experience, highlighting the importance of training. Clinicians acknowledged that the ADRe Profile was comprehensive but identified practical difficulties. Instruments to reduce ADRs should be validated before testing in feasibility studies and randomised controlled trials. Implications for Nursing Management: Managers need to optimise patient safety by introducing patient-centred symptom monitoring, with decision support. Before instruments are adopted, managers should check the reliability and validity data. Trial Registration: ClinicalTrials.gov identifier: NCT04663360.
- MeSH
- lidé středního věku MeSH
- lidé MeSH
- nežádoucí účinky léčiv * prevence a kontrola MeSH
- polypharmacy MeSH
- průzkumy a dotazníky MeSH
- psychometrie * přístrojové vybavení metody normy MeSH
- reprodukovatelnost výsledků MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- randomizované kontrolované studie MeSH
- validační studie MeSH
- Geografické názvy
- Spojené království MeSH
- MeSH
- ethanolamin analýza metabolismus MeSH
- finanční podpora výzkumu jako téma MeSH
- isochinoliny analýza metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- reprodukovatelnost výsledků MeSH
- řízení kvality MeSH
- techniky kombinatorické chemie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- srovnávací studie MeSH
The aim of this study was to optimise a two-tube reverse transcription triplex quantitative real time PCR (qRT-PCR) combining amplification of two loci with an internal amplification control (IAC) for detection and quantitation of hepatitis E virus (HEV) RNA and to validate its performance on a pool of biological samples. Optimisation was performed on serially diluted "home-made" RNA standards. The limit of detection was determined experimentally as 10 copies/μl of the RNA standard for both amplification targets. The qRT-PCR was validated on a cohort of samples originating from 48 wild boars (Sus scrofa), 17 fallow deer (Dama dama) and 28 mouflons (Ovis musimon), with nested RT-PCR used as a reference method. qRT-PCR was found to be more specific for the detection of HEV RNA in examined samples. HEV RNA was detected in samples of five more animals (one wild boar and four mouflons) in comparison with nested RT-PCR.
- MeSH
- hepatitida E diagnóza veterinární virologie MeSH
- ovce virologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody normy MeSH
- reprodukovatelnost výsledků MeSH
- RNA virová analýza genetika normy MeSH
- senzitivita a specificita MeSH
- Sus scrofa virologie MeSH
- virus hepatitidy E genetika MeSH
- vysoká zvěř virologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Comparison of HPLC methods using conventional particle-based and monolithic columns for determination of indomethacin and its two degradation products, viz. 5-methoxy-2-methylindoleacetic acid and 4-chlorobenzoic acid, was carried out. Ketoprofen was used as an internal standard for data evaluation throughout the study. Conventional separation was based on analytical column Zorbax SB-Phenyl (75x4.6 mm; 3.5 microm particles) used with a mobile phase composition of acetonitrile and phosphoric acid 0.2% (50:50, v/v) and isocratic flow at 0.6 mL/min. Three different lengths of Chromolith columns RP-18e (25x4.6 mm, 50x4.6 mm, and 100x3 mm) were tested with respect to the validation parameters peak asymmetry, resolution, height equivalent to a theoretical plate, repeatability, and after optimisation compared to values obtained using a conventional Zorbax column. The developed methods were used to determine all three compounds in a pharmaceutical formulation--Indobene gel. Chromatographic parameters were comparable to those of a conventional particle-based column. The analysis time was shortened as expected (retention times were lowered by a factor of two). Moreover, the repeatability of peak areas and retention times obtained with a 50 mm monolithic column was greatly improved (RSD values were lower than 0.40%).
The activities of 54Mn and 65Zn have been determined by 4pi(PC)-gamma coincidence counting, with efficiency variation performed by the conventional method of altering the self-absorption in the sources as well as by the computer discrimination method. The standardisation of 65Zn presents some complications requiring optimisation of the gamma-ray energy window settings to achieve a linear efficiency-extrapolation curve. Determination of these optimal settings by the conventional coincidence method is a tedious task. These difficulties have been reduced by the utilisation of a software coincidence counting system that records time and amplitude information of individual pulses from coincidence measurements, where the coincidence parameters are set after the data collection process has completed, facilitating multiple data evaluations on a single data set. The optimal gamma-ray energy window settings for the 65Zn standardisation were derived from the results of the 54Mn standardisation, as well as from studies of the 65Zn data itself. The setting of the PC channel thresholds for K and both (K+L) electrons is also discussed. The results are compared with those attained using conventional coincidence counting.
- MeSH
- algoritmy MeSH
- dávka záření MeSH
- mangan analýza normy MeSH
- radioizotopy zinku analýza normy MeSH
- radionuklidy analýza normy MeSH
- referenční hodnoty MeSH
- referenční standardy MeSH
- reprodukovatelnost výsledků MeSH
- scintilace - počítání metody normy MeSH
- senzitivita a specificita MeSH
- směrnice jako téma MeSH
- software MeSH
- spektrometrie gama metody normy MeSH
- Publikační typ
- hodnotící studie MeSH
- Geografické názvy
- Česká republika MeSH
An electrophoretic method (on-line coupled capillary isotachophoresis with capillary zone electrophoresis) with conductometric detection (cITP-CZE-COND) for the determination of histamine in foodstuffs and feedstuffs is described. Under optimised conditions in cationic mode the histamine is well separated from other components of acidic sample extract and detected by conductimeter within 25 min. Method characteristics, i.e., linearity (22-222 ng/mL), accuracy (recovery 91 ± 9%), repeatability (1.5%), reproducibility (3.4%), and detection limit (4 ng/mL) were evaluated. On a representative series of 37 food and feed samples it has been shown that the cITP-CZE-COND gives comparable results as routine accredited HPLC method. Low laboriousness, high sensitivity, speed of analysis, and low running cost are important attributes of cITP-CZE-COND.
A new CZE method was developed for the determination of 12 purine and pyrimidine nucleotides, two adenine coenzymes and their reduced forms, and acetyl coenzyme A in various cell extracts. As the concentration levels of these metabolites in living cells are low; CZE was combined with field-enhanced sample stacking. As a result, the separation conditions were optimised to achieve a suitable resolution at the relatively high sample volume provided by this on-line pre-concentration technique. The optimum BGE was 150 mM glycine buffer (pH 9.5). Samples were introduced hydrodynamically using a pressure of 35 mbar (3.5 kPa) for 25 s, and data were collected at a detection wavelength of 260 nm. An applied voltage of 30 kV (positive polarity) and capillary temperature of 25°C gave the best separation of these compounds. The optimised method was validated by determining the linearity, sensitivity and repeatability and it was successfully applied for the analysis of extracts from Paracoccus denitrificans bacteria and from stem cells.
- MeSH
- acetylkoenzym A analýza MeSH
- adenosintrifosfát analýza MeSH
- chemické techniky analytické metody normy MeSH
- cytidintrifosfát analýza MeSH
- embryonální kmenové buňky chemie MeSH
- guanosintrifosfát analýza MeSH
- lidé MeSH
- limita detekce MeSH
- Paracoccus denitrificans chemie MeSH
- reprodukovatelnost výsledků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A CE method with contactless conductivity detection has been developed for the clinical determination of the branched chain amino acids (BCAAs) valine, isoleucine and leucine in human blood plasma. The CE separation was performed in an optimised BGE with composition of 3.2 M acetic acid in 20% v/v methanol, pH 2.0. The achieved separation time was 125 s when using a capillary with an effective length of 14.7 cm, electric field intensity of 0.96 kV/cm and simultaneous application of a hydrodynamic pressure of 50 mbar. The separation efficiency in blood plasma equalled 461 000 theoretical plates/m for valine and isoleucine, and 455 000 theoretical plates/m for leucine; the detection limits are equal to 0.4 μM for all three amino acids. The RSD values for repeatability of the migration time equalled 0.1% for measurements during a single day and 0.3% for measurements on different days; the RSD values for repeatability of the peak areas equalled 2.3-2.6% for measurements during a single day and 2.7-4.6% for measurements on different days. It followed from the performed tests that the plasmatic levels of BCAAs attain a maximum 60 min after intravenous application of an infusion of BCAAs.
- MeSH
- diabetes mellitus MeSH
- dospělí MeSH
- elektrická vodivost MeSH
- elektroforéza kapilární metody MeSH
- inzulin krev metabolismus MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- limita detekce MeSH
- lineární modely MeSH
- reprodukovatelnost výsledků MeSH
- větvené aminokyseliny krev farmakokinetika MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The triterpenoid plant hormones brassinosteroids (BRs) are believed to influence almost every aspect of plant growth and development. We have developed a sensitive mass spectrometry-based method for the simultaneous profiling of twenty-two naturally occurring brassinosteroids including biosynthetic precursors and the majority of biologically active metabolites. Using ultra-high performance liquid chromatographic (UHPLC) analysis, the run time was reduced up to three times (to 9 min) in comparison to standard HPLC BRs analyses, the retention time stability was improved to 0.1-0.2 % RSD and the injection accuracy was increased to 1.1-4.9 % RSD. The procedures for extraction and for two-step purification based on solid-phase extraction (SPE) were optimised in combination with subsequent UHPLC analysis coupled to electrospray ionisation tandem mass spectrometry (ESI-MS/MS) using Brassica flowers and Arabidopsis plant tissue extracts. In multiple reaction monitoring (MRM) mode, the average detection limit for BRs analysed was close to 7 pg, and the linear range covered up to 3 orders of magnitude. The low detection limits for this broad range of BR metabolites enabled as little as 50 mg of plant tissue to be used for quantitative analyses. The results of determinations exploiting internal standards showed that this approach provides a high level of practicality, reproducibility and recovery. The method we have established will enable researchers to gain a better understanding of the dynamics of the biosynthesis and metabolism of brassinosteroids and their modes of action in plant growth and development.
- MeSH
- Arabidopsis chemie MeSH
- Brassica napus chemie MeSH
- brassinosteroidy analýza MeSH
- extrakce na pevné fázi metody MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- limita detekce MeSH
- reprodukovatelnost výsledků MeSH
- rostlinné extrakty chemie MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- validační studie MeSH