OBJECTIVES: Stability of concentrations of urinary stone-related metabolites was analyzed from samples of recurrent urinary stone formers to assess necessity and effectiveness of urine acidification during collection and storage. METHODS: First-morning urine was collected from 20 adult calcium-stone forming patients at Tomas Bata Hospital in the Czech Republic. Urine samples were analyzed for calcium, magnesium, inorganic phosphate, uric acid, sodium, potassium, chloride, citrate, oxalate, and urine particles. The single-voided specimens were collected without acidification, after which they were divided into three groups for storage: samples without acidification ("NON"), acidification before storage ("PRE"), or acidification after storage ("POST"). The analyses were conducted on the day of arrival (day 0, "baseline"), or after storage for 2 or 7 days at room temperature. The maximum permissible difference (MPD) was defined as ±20 % from the baseline. RESULTS: The urine concentrations of all stone-related metabolites remained within the 20 % MPD limits in NON and POST samples after 2 days, except for calcium in NON sample of one patient, and oxalate of three patients and citrate of one patient in POST samples. In PRE samples, stability failed in urine samples for oxalate of three patients, and for uric acid of four patients after 2 days. Failures in stability often correlated with high baseline concentrations of those metabolites in urine. CONCLUSIONS: Detailed procedures are needed to collect urine specimens for analysis of urinary stone-related metabolites, considering both patient safety and stability of those metabolites. We recommend specific preservation steps.
- MeSH
- analýza moči metody MeSH
- dospělí MeSH
- koncentrace vodíkových iontů MeSH
- kyselina močová moč MeSH
- lidé středního věku MeSH
- lidé MeSH
- močové kameny * moč MeSH
- odběr biologického vzorku metody MeSH
- recidiva * MeSH
- sbírání vzorku moči metody MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- antibiotická rezistence MeSH
- antigeny analýza MeSH
- diagnostické techniky dýchacího ústrojí klasifikace MeSH
- diagnostické techniky molekulární MeSH
- infekce dýchací soustavy * diagnóza etiologie mikrobiologie MeSH
- kultivační techniky metody MeSH
- lidé MeSH
- mikrobiologické techniky * klasifikace metody MeSH
- mikroskopie metody MeSH
- odběr biologického vzorku metody MeSH
- odběr vzorku krve MeSH
- protilátky analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
The MBT Pathfinder is an automated colony-picking robot designed for efficient sample preparation in matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. This article presents results from three key experiments evaluating the instrument's performance in conjunction with MALDI Biotyper instrument. The method comparison experiment assessed its clinical performance, demonstrating comparable results with gram-positive, gram-negative, and anaerobic bacteria (scores larger than 2.00) and superior performance over simple direct yeast transfer (score: 1.80) when compared to samples prepared manually. The repeatability experiment confirmed consistent performance over multiple days and labs (average log score: 2.12, std. deviation: 0.59). The challenge panel experiment showcased its consistent and accurate performance across various samples and settings, yielding average scores between 1.76 and 2.19. These findings underline the MBT Pathfinder as a reliable and efficient tool for MALDI-TOF mass spectrometry sample preparation in clinical and research applications.
- MeSH
- Bacteria * klasifikace izolace a purifikace MeSH
- laboratorní automatizace * metody MeSH
- lidé MeSH
- odběr biologického vzorku metody MeSH
- reprodukovatelnost výsledků MeSH
- robotika * MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice * metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
Osteosarcoma and Ewing sarcoma are bone tumors mostly diagnosed in children, adolescents, and young adults. Despite multimodal therapy, morbidity is high and survival rates remain low, especially in the metastatic disease setting. Trials investigating targeted therapies and immunotherapies have not been groundbreaking. Better understanding of biological subgroups, the role of the tumor immune microenvironment, factors that promote metastasis, and clinical biomarkers of prognosis and drug response are required to make progress. A prerequisite to achieve desired success is a thorough, systematic, and clinically linked biological analysis of patient samples, but disease rarity and tissue processing challenges such as logistics and infrastructure have contributed to a lack of relevant samples for clinical care and research. There is a need for a Europe-wide framework to be implemented for the adequate and minimal sampling, processing, storage, and analysis of patient samples. Two international panels of scientists, clinicians, and patient and parent advocates have formed the Fight Osteosarcoma Through European Research consortium and the Euro Ewing Consortium. The consortia shared their expertise and institutional practices to formulate new guidelines. We report new reference standards for adequate and minimally required sampling (time points, diagnostic samples, and liquid biopsy tubes), handling, and biobanking to enable advanced biological studies in bone sarcoma. We describe standards for analysis and annotation to drive collaboration and data harmonization with practical, legal, and ethical considerations. This position paper provides comprehensive guidelines that should become the new standards of care that will accelerate scientific progress, promote collaboration, and improve outcomes.
- MeSH
- banky biologického materiálu MeSH
- Ewingův sarkom * terapie patologie diagnóza MeSH
- lidé MeSH
- nádorové biomarkery MeSH
- nádory kostí * terapie patologie MeSH
- odběr biologického vzorku * metody normy MeSH
- osteosarkom * terapie patologie diagnóza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Geografické názvy
- Evropa MeSH
- MeSH
- antifungální látky aplikace a dávkování farmakologie terapeutické užití MeSH
- dermatomykózy diagnóza etiologie farmakoterapie MeSH
- kandidóza diagnóza etiologie farmakoterapie MeSH
- lidé MeSH
- mykózy * diagnóza etiologie farmakoterapie MeSH
- oční infekce mykotické diagnóza etiologie farmakoterapie MeSH
- odběr biologického vzorku metody MeSH
- otomykóza diagnóza etiologie farmakoterapie MeSH
- plicní mykózy diagnóza etiologie farmakoterapie MeSH
- rizikové faktory MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Proper fixing and long-term preservation of entomological evidence are essential in collections and research and crucial in applied fields such as forensic entomology. Incorrectly stored samples may lose important morphological features over time, rendering molecular analyses exceedingly difficult. The most effective method for preserving soft samples such as larvae is fluid preservation. It uses a combination of a wide range of fixatives and storage fluids. However, very little comparative work has been done to determine the effects of long-term storage on sample quality in terms of color, shape, and DNA stability. Moreover, the current golden standard in forensic entomology has been tailored for age estimation of larvae of Diptera, which differ from larvae of Coleoptera in morphology and subsequently in applied methods. We compared the effects of combinations of 6 commonly used fixatives and 6 commonly used storage fluids on midsized larvae of the forensically important beetle, Necrodes littoralis (Linnaeus, 1758), in terms of color, shape, and suitability for DNA analyses over a 2-yr period. We were looking for combinations that can preserve specimens in a satisfactory state, can be used on a regular basis, do not require advanced protection or skills of the personnel, and are not toxic or too harmful to the environment. We found not only several methods that scored significantly better in the tested parameters compared with the golden standard but also several common methods that should be avoided. The effects of agents on each tested category are discussed in detail.
- MeSH
- barva MeSH
- brouci * MeSH
- časové faktory MeSH
- DNA * analýza MeSH
- forenzní entomologie metody MeSH
- larva * růst a vývoj MeSH
- ochrana biologická metody MeSH
- odběr biologického vzorku metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
BACKGROUND: Testing of pooled samples is an effective strategy for increasing testing capacity while saving resources and time. This study aimed to validate pooled testing and gather real-life data on its use for Covid-19 surveillance with a gargle lavage (GL) self-sampling strategy. METHODS: Two-stage pooled testing with pools of 6 and 12 samples was used for preventive testing of an asymptomatic population and Covid-19 surveillance in Czech schools. Both GL and nasopharyngeal swabs were used for sampling. RESULTS: In total, 61,111 samples were tested. The use of pooled testing for large-scale Covid-19 surveillance reduced consumable costs by almost 75% and increased testing capacity up to 3.8-fold compared to standard methods. RT-PCR experiments revealed a minimal loss of sensitivity (0-2.2%) when using pooled samples, enabling the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genes with Ct values >35. The minor loss of sensitivity was counterbalanced by a significantly increased throughput and the ability to substantially increase testing frequencies. CONCLUSIONS: Pooled testing is considerably more cost-effective and less time-consuming than standard testing for large-scale Covid-19 surveillance even when the prevalence of SARS-CoV-2 is fluctuating. Gargle lavage self-sampling is a non-invasive technique suitable for sample collection without a healthcare worker's assistance.
- MeSH
- COVID-19 * diagnóza epidemiologie MeSH
- lidé MeSH
- nazofarynx * virologie MeSH
- odběr biologického vzorku * metody MeSH
- SARS-CoV-2 * genetika izolace a purifikace MeSH
- senzitivita a specificita MeSH
- testování na COVID-19 průkazem nukleové kyseliny metody MeSH
- testování na COVID-19 metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
One of the least invasive sampling methods suitable for self-sampling is saliva spitting. The aim of this study is to evaluate the suitability of saliva self-sampling for unsupervised testing. Two self-sampling strategies were compared on the basis of visual evaluation of samples, measurement of cortisol levels in samples and questionnaire survey. The saliva samples obtained by supervised self-sampling were found to be fully suitable for further analysis. In contrast, not all saliva samples obtained from unsupervised self-collection can be used: 13% non-compliance with the minimum required sample volume, 8% with some food/drink residues and 26% taken at the wrong day time. About 42% of the unsupervised probands made at least one significant error in the saliva self-collection procedure. These results indicate that the accuracy of the results based on the analysis of samples received from saliva self-sampling is limited. For clinical investigation, the presence of an inner standard (referring to the reliability of the sampling procedure) is required.
One of the challenges in clinical translation of cell-replacement therapies is the definition of optimal cell generation and storage/recovery protocols which would permit a rapid preparation of cell-treatment products for patient administration. Besides, the availability of injection devices that are simple to use is critical for potential future dissemination of any spinally targeted cell-replacement therapy into general medical practice. Here, we compared the engraftment properties of established human-induced pluripotent stem cells (hiPSCs)-derived neural precursor cell (NPCs) line once cells were harvested fresh from the cell culture or previously frozen and then grafted into striata or spinal cord of the immunodeficient rat. A newly developed human spinal injection device equipped with a spinal cord pulsation-cancelation magnetic needle was also tested for its safety in an adult immunosuppressed pig. Previously frozen NPCs showed similar post-grafting survival and differentiation profile as was seen for freshly harvested cells. Testing of human injection device showed acceptable safety with no detectable surgical procedure or spinal NPCs injection-related side effects.
- MeSH
- buněčná diferenciace fyziologie MeSH
- dospělí MeSH
- genetické vektory genetika MeSH
- indukované pluripotentní kmenové buňky * fyziologie transplantace MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- mícha MeSH
- mozek MeSH
- nervové kmenové buňky * fyziologie transplantace MeSH
- odběr biologického vzorku metody MeSH
- odběr tkání a orgánů metody MeSH
- prasata MeSH
- přeprogramování buněk * genetika fyziologie MeSH
- přežívání štěpu fyziologie MeSH
- spinální injekce * škodlivé účinky přístrojové vybavení metody MeSH
- transplantace kmenových buněk * škodlivé účinky přístrojové vybavení metody MeSH
- virus Sendai MeSH
- výsledek terapie MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- biologické markery * analýza MeSH
- chybná diagnóza MeSH
- diagnostické techniky gastrointestinální klasifikace trendy MeSH
- ELISA MeSH
- feces * mikrobiologie MeSH
- gastrointestinální nemoci diagnostické zobrazování klasifikace MeSH
- lidé MeSH
- metody pro přípravu analytických vzorků metody MeSH
- odběr biologického vzorku metody MeSH
- pracovníci laboratoře MeSH
- prediktivní hodnota testů MeSH
- senzitivita a specificita MeSH
- Check Tag
- lidé MeSH
