We report the forensic case of a 42-year-old man, a known drug user, who died at home and whose body was only discovered 2 months later. Autopsy was performed on a corpse in the late postmortem stage where no apparent cause of death was found. A toxicological screening of biological materials (blood, urine and gastric content) using liquid chromatography with different types of mass detection (ion trap and high-resolution) revealed the presence of methoxetamine (MXE), a ketamine analog, and its metabolites. MXE and a number of its metabolites (e.g., O-desmethyl, N-desethyl, hydroxy, glucuronides and sulfates) were identified in urine. Based on the results, a method using liquid chromatography with tandem mass spectrometry was developed and validated for the determination of MXE concentration in biological materials. The following values of MXE concentration were found: blood-3.6 ng/mL, urine-70.5 ng/mL and gastric content-18.0 ng/mL. Given the absence of other drugs, medications and poisons, it can be inferred that despite relatively low blood concentrations, MXE contributed to the victim's death. The present case demonstrates that even after 2 months, MXE and its several metabolites can be detected and determined in the human cadaver at a relatively advanced stage of decomposition.
- MeSH
- cyklohexanony * analýza metabolismus MeSH
- cyklohexylaminy * MeSH
- dospělí MeSH
- lidé MeSH
- mrtvola MeSH
- pitva MeSH
- tělesné tekutiny * metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
MicroRNAs (miRNAs) comprise a large group of small noncoding RNAs within a heterogeneous entity of noncoding RNAs, forming potent functional tools regulating the crucial biological processes within cells and the body. Cell-free miRNAs have become one of the novel promising diagnostic, predictive, and prognostic biomarkers for various diseases extensively investigated in recent years. This is due to their presence within extracellular fractions of various body fluids suggesting their potential as noninvasive "liquid biopsy" in case of their dysregulated expression.Among the body fluids, blood plasma and serum along with urine are the most commonly investigated sources of various types of cell-free miRNAs. Another body fluid, i.e., ascites (effusion, peritoneal/pleural fluid) may be the clinically important fluid particularly associated with carcinogenesis in ovarian carcinomas and hepatocellular carcinomas or in case of liver cirrhosis.Here, we provide a protocol for an expression profiling study based on qPCR analyses aimed at finding novel candidate miRNAs via small-scale or large-scale screening and evaluation experiments using liquid biopsies of blood plasma, ascites, and urine. Using this approach may be worth in cases where no (or limited) information is available on miRNA expression in particular diseases and geographic regions, for validation of previously published miRNAs with promising diagnostic potential, particularly in situations where follow-up study is aimed at validating miRNAs coming from (micro) array or NGS experiments, or where funding for large-scale experiments is not available. We demonstrate that assessment of plasma, ascites, and urine miRNAs expression may represent a feasible method to explore the potential for finding novel diagnostic, predictive, and prognostic biomarkers for various diseases.
Extracellular RNAs present in biofluids have emerged as potential biomarkers for disease. Where most studies focus on blood-derived fluids, other biofluids may be more informative. We present an atlas of messenger, circular, and small RNA transcriptomes of a comprehensive collection of 20 human biofluids. By means of synthetic spike-in controls, we compare RNA content across biofluids, revealing a 10,000-fold difference in concentration. The circular RNA fraction is increased in most biofluids compared to tissues. Each biofluid transcriptome is enriched for RNA molecules derived from specific tissues and cell types. Our atlas enables an informed selection of the most relevant biofluid to monitor particular diseases. To verify the biomarker potential in these biofluids, four validation cohorts representing a broad spectrum of diseases were profiled, revealing numerous differential RNAs between case and control subjects. Spike-normalized data are publicly available in the R2 web portal for further exploration.
- MeSH
- biologické markery * MeSH
- kohortové studie MeSH
- lidé MeSH
- RNA genetika metabolismus MeSH
- sekvenční analýza RNA metody MeSH
- stanovení celkové genové exprese metody MeSH
- tělesné tekutiny metabolismus MeSH
- transkriptom * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
The international standard ISO 23317:2014 for the in vitro testing of inorganic biomaterials in simulated body fluid (SBF) uses TRIS buffer to maintain neutral pH. In our previous papers, we investigated the interaction of a glass-ceramic scaffold with TRIS and HEPES buffers. Both of them speeded up glass-ceramic dissolution and hydroxyapatite (HAp) precipitation, thereby demonstrating their unsuitability for the in vitro testing of highly reactive biomaterials. In this article, we tested MOPS buffer (3-[N-morpholino] propanesulfonic acid), another amino acid from the group of "Goods buffers". A highly reactive glass-ceramic scaffold (derived from Bioglass®) was exposed to SBF under static-dynamic conditions for 13/15 days. The kinetics and morphology of the newly precipitated HAp were studied using two different concentrations of (PO4 )3- ions in SBF. The pH value and the SiIV , Ca2+ , and (PO4 )3- concentrations in the SBF leachate samples were measured every day (AAS, spectrophotometry). The glass-ceramic scaffold was monitored by SEM/EDS, XRD, WD-XRF, and BET before and after 1, 3, 7, 11, and 13/15 days of exposure. As in the case of TRIS and HEPES, the preferential dissolution of the glass-ceramic crystalline phase (Combeite) was observed, but less intensively. The lower concentration of (PO4 )3- ions slowed down the kinetics of HAp precipitation, thereby causing the disintegration of the scaffold structure. This phenomenon shows that the HAp phase was predominately generated by the presence of (PO4 )3- ions in the SBF, not in the glass-ceramic material. Irrespective of this, MOPS buffer is not suitable for the maintenance of pH in SBF.
The release of neutrophil extracellular traps (NETs) is one of the weapons neutrophils have in their armory. NETs consist of extracellular chromatin fibers decorated with a plethora of cytoplasmic and granular proteins, such as the antimicrobial serine protease neutrophil elastase (NE). Because the first description of NETs as beneficial to the host, reports on their double-faced role in health and disease have considerably increased recently. On one hand, NETs reportedly trap and kill bacteria and also participate in the resolution of the acute inflammation associated with infection and with tissue damage. On the other hand, numerous negative aspects of NETs contribute to the etiopathogenesis of autoimmune disorders. Employing soluble and solid fluorescent substrates, we demonstrate the interaction of NE with aggregated NETs (aggNETs), the limitation of its enzymatic activity and the containment of the enzyme from surrounding tissues. These events prevent the spread of inflammation and tissue damage. The detection of DNase 1-dependent elevation of NE activity attests the continuous presence of patrolling neutrophils forming NETs and aggNETs even under conditions physiologic conditions.
- MeSH
- aktivace enzymů MeSH
- deoxyribonukleasa I metabolismus MeSH
- deoxyribonukleasy metabolismus MeSH
- extracelulární pasti imunologie metabolismus MeSH
- leukocytární elastasa metabolismus MeSH
- lidé MeSH
- myši MeSH
- neutrofily imunologie metabolismus MeSH
- tělesné tekutiny metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Progress achieved between 2014-2017 in the extraction and sample preparation of nucleic acid by isotachophoresis is reviewed in this paper. The isolation and purification of nucleic acids is very often compromised by a complex matrix such as blood and other bodily fluids, samples from the scene of crime, fossil samples, etc. While most of the common nucleic acids isolation techniques are based on extraction with inherent limitations with regard to quantitative results, isotachophoretic focusing is a quantitative process with a theoretically unlimited concentration factor. Since isotachophoresis belongs to less traditional approaches of nucleic acids purification, we present not only the latest developments in the application of isotachophoresis for the nucleic acids concentration but also a brief description of the principles of this method.
- MeSH
- automatizace MeSH
- DNA analýza MeSH
- elektrolyty MeSH
- hybridizace nukleových kyselin MeSH
- izotachoforéza metody trendy MeSH
- lidé MeSH
- mikro RNA analýza MeSH
- mikrofluidika MeSH
- nukleové kyseliny izolace a purifikace MeSH
- polymerázová řetězová reakce MeSH
- pufry MeSH
- tělesné tekutiny metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Změny objemu i složení tělesných tekutin patří mezi zásadní a limitující parametry ve zdraví i v nemoci. S přibývajícím věkem význam těchto parametrů nabývá na závažnosti. V konceptu geriatrického pacienta jsou rozvraty vodního a minerálního metabolizmu příčinou oběhového selhání, centrálních mozkových příhod, dále instability, pádů, deliria. Organizmus sice umí v širokém rozmezí rovnováhy ve vnitřním prostředí odchylky kompenzovat, ale vždy jen po limitovanou dobu, a zejména ve stáří tato schopnost kompenzace klesá. Udržení vodní a minerální rovnováhy ve stáří je komplikováno i polymorbiditou. Častý výskyt kardiovaskulárních chorob a omezení renálních funkcí ve stáří má za následek omezené kompenzační schopnosti a tento stav je nutné striktně respektovat. Vedle polymorbidity se ve stáří uplatňuje při vzniku poruch vodního a elektrolytového hospodářství i velmi častá polypragmazie ve farmakoterapii. Sedativní léčba potlačuje i pocit žízně, a to má za následek neobvykle rychlý rozvoj poruch vodní a minerální rovnováhy i po malých inzultech, jako jsou horečnatá onemocnění a drobné úrazy. Znalost odlišností diagnostiky a léčby vodních a iontových dysbalancí ve stáří nabývá stále větší důležitosti, zejména při rostoucím podílu starší populace ve společnosti.
Changes in the volume and composition of body fluids are among the essential and limiting parameters both in health and illness. These parameters gain in importance with increasing age. Within the concept of a geriatric patient, disturbances in water and mineral metabolism are the cause of circulatory collapse, stroke, and further instability, falls and delirium. The body can, in the broad range of balance within internal environment, compensate for variations, however always for a limited length of time only, and this compensation ability decreases in particular in older age. Maintaining of water and mineral balance in the elderly is also complicated by polymorbidity. Frequent occurrence of cardiovascular diseases and decline in renal functions later in life results in reduced compensation abilities, which status must be rigorously considered. Besides polymorbidity, also polypragmasia in pharmacotherapy is very frequently encountered in relation to age-related disorders of water and electrolyte handling. Treatment with sedatives also suppresses the feeling of thirst, which results in rapid development of disturbances in water and mineral balance even after small insults, such as feverish illnesses and minor injuries. The knowledge of differences in diagnosing and treatment of water and ion imbalances in later life is becoming increasingly important, especially with regard to the increasing share of older people in society.
- MeSH
- dehydratace MeSH
- hořčík krev metabolismus MeSH
- hyperkalemie etiologie patofyziologie terapie MeSH
- hypernatremie etiologie terapie MeSH
- hypokalemie etiologie chemicky indukované patologie terapie MeSH
- lidé MeSH
- poruchy metabolismu fosforu etiologie klasifikace patologie terapie MeSH
- poruchy metabolismu vápníku etiologie klasifikace patologie terapie MeSH
- senioři MeSH
- stárnutí MeSH
- tělesné tekutiny metabolismus MeSH
- věkové faktory MeSH
- vodní a elektrolytová nerovnováha * etiologie klasifikace patofyziologie terapie MeSH
- vodní a elektrolytová rovnováha účinky léků MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
OBJECTIVE: To determine the cervical and vaginal fluid soluble Toll-like receptor-2 (sTLR2) levels in pregnancies complicated by preterm prelabor rupture of membranes (PPROM) and their correlation to microbial invasion of the amniotic cavity (MIAC) and/or histological chorioamnionitis (HCA). METHODS: Sixty-eight women with singleton pregnancies complicated by PPROM were included in this study. Cervical and vaginal fluid was collected at the time of admission, and levels of sTLR2 in the cervical and vaginal fluid were determined using enzyme-linked immunosorbent assay. RESULT: Women with MIAC and both MIAC and HCA did not have different cervical and vaginal fluid sTLR2 levels compared to those without MIAC and without both MIAC and HCA. Women with HCA had higher cervical fluid sTLR2 levels in crude analysis (with HCA: median 11.6 pg/mL versus without HCA: median 5.5 pg/mL; p = 0.04) but not after adjustment for gestational age at sampling (p = 0.19). No difference in vaginal fluid sTLR2 levels between women with and without HCA was found. A positive correlation between cervical and vaginal fluid sTLR2 levels was identified (rho = 0.54; p < 0.0001). CONCLUSIONS: Cervical and vaginal fluid sTLR2 levels did not reflect the presence of MIAC and/or HCA.
- MeSH
- cervix uteri metabolismus MeSH
- chorioamnionitida metabolismus MeSH
- dospělí MeSH
- ELISA MeSH
- lidé MeSH
- novorozenec MeSH
- plodová voda mikrobiologie MeSH
- předčasný odtok plodové vody metabolismus patologie MeSH
- těhotenství MeSH
- tělesné tekutiny metabolismus MeSH
- toll-like receptor 2 metabolismus MeSH
- vagina metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- novorozenec MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVE: To determine the vaginal fluid interleukin (IL)-6 and IL-8 concentrations in pregnancies complicated by preterm prelabor rupture of membranes and their correlation to microbial invasion of the amniotic cavity (MIAC) as well as histological chorioamnionitis (HCA). METHODS: Sixty-eight women with singleton pregnancies were included in this study. Vaginal fluid was collected at the time of admission. IL-6 and IL-8 concentrations in the vaginal fluid were determined using ELISA. RESULT: Women with MIAC had higher vaginal fluid IL-6 levels compared to those without MIAC (with MIAC: median 374 pg/mL versus without MIAC: median 174 pg/mL; p = 0.03). IL-8 levels were higher in women with MIAC only in the crude analysis but not after adjustment for gestational age. There was no difference in the IL-6 and IL-8 concentrations between those with and without HCA. Women with both MIAC and HCA had higher IL-6 vaginal fluid levels than those without both MIAC and HCA (with MIAC and HCA: median 466 pg/mL versus without MIAC and HCA: median 178 pg/mL; p = 0.02). IL-8 levels were higher in women with MIAC and HCA only in the crude analysis but not after adjustment for gestational age. CONCLUSIONS: Vaginal fluid IL-6 but not IL-8 levels reflect the presence of MIAC and both MIAC and HCA.
- MeSH
- chorioamnionitida metabolismus patologie MeSH
- dospělí MeSH
- infekční komplikace v těhotenství metabolismus patologie MeSH
- interleukin-6 analýza metabolismus MeSH
- interleukin-8 analýza metabolismus MeSH
- kohortové studie MeSH
- lidé MeSH
- mladý dospělý MeSH
- novorozenec MeSH
- předčasný odtok plodové vody metabolismus patologie MeSH
- těhotenství MeSH
- tělesné tekutiny chemie metabolismus MeSH
- vagina MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- novorozenec MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
UNLABELLED: Abstract Objective: To determine the cervical fluid interleukin (IL)-6 and IL-8 levels in pregnancies complicated by preterm prelabor rupture of membranes (PPROM) and the association of these interleukins with microbial invasion of the amniotic cavity (MIAC) and histological chorioamnionitis (HCA). METHODS: Sixty women with singleton pregnancies were included in this study. Cervical fluid was sampled at the time of admission using Dacron polyester swabs, which were placed into the endocervical canal for 20 s. IL-6 and IL-8 levels were determined by ELISA. The management of PPROM was active management (except for in pregnancies <28 weeks of gestation) and occurs not later than 72 h after the rupture of membranes. RESULT: The women with MIAC had higher IL-6 and IL-8 levels than did the women without MIAC (IL-6: p=0.01; IL-8: p=0.003). There was no difference in IL-6 levels between women with and without HCA (p=0.37). The women with HCA had higher IL-8 levels only in the crude analysis (p=0.01) but not after adjustment for gestational age (p=0.06). The women with both MIAC and HCA had higher levels of IL-6 and IL-8 than did the other women (IL-6: p=0.003; IL-8: p=0.001). IL-8 level of 2653 pg/mL was found to be the best cut-off point in the identification of PPROM pregnancies complicated by both MIAC and HCA with a likelihood ratio of 24. CONCLUSIONS: The presence of MIAC is the most important factor impacting the local cervical inflammatory response, which is determined by IL-6 and IL-8 levels in the cervical fluid. IL-8 levels seem to be a promising non-invasive marker for the prediction of pregnancies complicated by the presence of both MIAC and HCA.
- MeSH
- amnion metabolismus mikrobiologie patologie MeSH
- cervix uteri chemie metabolismus MeSH
- chorioamnionitida diagnóza metabolismus mikrobiologie patologie MeSH
- dospělí MeSH
- infekční komplikace v těhotenství diagnóza metabolismus MeSH
- interleukin-6 analýza metabolismus MeSH
- interleukin-8 analýza metabolismus MeSH
- lidé MeSH
- mladý dospělý MeSH
- plodová voda chemie metabolismus MeSH
- předčasný odtok plodové vody metabolismus MeSH
- těhotenství MeSH
- tělesné tekutiny chemie metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
