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
• Názvy látek
• 2-(3-methoxyphenyl)-2-(ethylamino)cyclohexanone MeSH Prohlížeč
• cyklohexanony * MeSH
• cyklohexylaminy * 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.

• Klíčová slova
• Ascites, Biomarker, Cancer, diagnostics, MicroRNA, Plasma, Real-time PCR, Urine,
• MeSH
• ascites MeSH
• biologické markery MeSH
• cirkulující mikroRNA * MeSH
• krevní plazma chemie   MeSH
• lidé MeSH
• mikro RNA * genetika   MeSH
• nádorové biomarkery MeSH
• následné studie MeSH
• tělesné tekutiny * metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• cirkulující mikroRNA * MeSH
• mikro RNA * MeSH
• nádorové biomarkery MeSH

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.

• Klíčová slova
• RNA sequencing, biofluids, biomarker, cell-free RNA, circular RNA, extracellular RNA, liquid biopsy, messenger RNA, small RNA,
• 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
• Názvy látek
• biologické markery * MeSH
• RNA 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.

• Klíčová slova
• MOPS buffer, bioglass®, glass-ceramics scaffold, in vitro test, simulated body fluid,
• MeSH
• difrakce rentgenového záření MeSH
• fosfáty chemie   MeSH
• hydroxyapatit chemie   MeSH
• ionty chemie   MeSH
• keramika chemie   MeSH
• kinetika MeSH
• koncentrace vodíkových iontů MeSH
• pufry MeSH
• tělesné tekutiny chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Bioglass MeSH Prohlížeč
• fosfáty MeSH
• Glass ceramics MeSH Prohlížeč
• hydroxyapatit MeSH
• ionty MeSH
• pufry MeSH

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.

• Klíčová slova
• DNase 1, NETs, aggregated NETs, neutrophil elastase, neutrophils,
• 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
• Názvy látek
• deoxyribonukleasa I MeSH
• deoxyribonukleasy MeSH
• leukocytární elastasa 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.

• Klíčová slova
• isotachophoresis, nucleic acids, sample preparation,
• 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
• Názvy látek
• DNA MeSH
• elektrolyty MeSH
• mikro RNA MeSH
• nukleové kyseliny MeSH
• pufry 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.

• Klíčová slova
• Intrauterine inflammation, local inflammatory response, preterm delivery,
• 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
• Názvy látek
• toll-like receptor 2 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.

• Klíčová slova
• Histological chorioamnionitis, microbial invasion of the amniotic cavity, preterm delivery,
• 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
• Názvy látek
• IL6 protein, human MeSH Prohlížeč
• interleukin-6 MeSH
• interleukin-8 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.

• Klíčová slova
• Chorioamnionitis, inflammatory response, non-invasive sample,
• 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
• Názvy látek
• CXCL8 protein, human MeSH Prohlížeč
• IL6 protein, human MeSH Prohlížeč
• interleukin-6 MeSH
• interleukin-8 MeSH

An infection of any part of female reproductive tract can severely interfere with fertility and reproduction. The fluids and epithelium from the lumen of the female reproductive tract (uterus, oviduct and ovarian follicle) are a known source of antimicrobial action in several species. In this study, we compared the antimicrobial properties of fluids from the reproductive tract of a cow. After removal of small molecules, we demonstrated that there is an antimicrobial activity connected with a fraction of compounds with a molecular mass range between 3500 and 30,000. The most probable candidates responsible for the observed antimicrobial effect were subsequently identified by mass spectroscopy as histones H2A type 2-C, H2B type 1-K, H3.3, and H4. The antimicrobial role of histone H2B was further confirmed by using an antibody against this histone.

• Klíčová slova
• Antimicrobial properties, Bovine reproductive tract, Extracellular histones, Histone-like proteins,
• MeSH
• antiinfekční látky metabolismus   MeSH
• dialýza MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• histony chemie   metabolismus   MeSH
• hmotnostní spektrometrie MeSH
• skot MeSH
• tělesné tekutiny metabolismus   MeSH
• vejcovody u zvířat metabolismus   MeSH
• ženské pohlavní orgány metabolismus   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antiinfekční látky MeSH
• histony MeSH