BACKGROUND: Understanding the temporal variability of the microbiome is critical for translating associations of the microbiome with health and disease into clinical practice. The aim of this study is to assess the extent of temporal variability of the human urinary microbiota. A pair of urine samples were collected from study participants at 3-40-month interval. DNA was extracted and the bacterial V4 hypervariable region of the 16S rRNA gene was sequenced on the Illumina MiSeq platform. The alpha diversity of paired samples was analyzed using Chao1 and Shannon indices and PERMANOVA was used to test the factors influencing beta diversity. RESULTS: A total of 63 participants (43 men and 20 women with a mean age of 63.0 and 57.1 years, respectively) were included in the final analysis. An average of 152 ± 128 bacterial operational taxonomic units (OTUs) were identified in each urine sample from the entire cohort. There was an average of 41 ± 32 overlapping OTUs in each sample pair, accounting for 66.3 ± 29.4% of the relative abundance. There was a clear correlation between the number of overlapping OTUs and the relative abundance covered. The difference in Chao1 index between paired samples was statistically significant; the difference in Shannon index was not. Beta diversity did not differ significantly within the paired samples. Neither age nor sex of the participants influenced the variation in community composition. With a longer interval between the collections, the relative abundance covered by the overlapping OTUs changed significantly but not the number of OTUs. CONCLUSION: Our findings demonstrated that, while the relative abundance of dominant bacteria varied, repeated collections generally shared more than 60% of the bacterial community. Furthermore, we observed little variation in the alpha and beta diversity of the microbial community in human urine. These results help to understand the dynamics of human urinary microbiota and enable interpretation of future studies.

• Klíčová slova
• 16S rRNA gene, Bacterial community, Next-generation sequencing, Stability, Urinary microbiota, Variability,
• MeSH
• Bacteria * klasifikace   genetika   izolace a purifikace   MeSH
• biodiverzita MeSH
• časové faktory MeSH
• DNA bakterií genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikrobiota * genetika   MeSH
• moč * mikrobiologie   MeSH
• prospektivní studie MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• Názvy látek
• DNA bakterií MeSH
• RNA ribozomální 16S MeSH

Capillary electrophoresis coupled online with mass detection is a modern tool for analyzing wide ranges of compounds in complex samples, including urine. Capillary electrophoresis with mass spectrometry allows the separation and identification of various analytes spanning from small ions to high molecular weight protein complexes. Similarly to the much more common liquid chromatography-mass spectrometry techniques, the capillary electrophoresis separation reduces the complexity of the mixture of analytes entering the mass spectrometer resulting in reduced ion suppression and a more straightforward interpretation of the mass spectrometry data. This review summarizes capillary electrophoresis with mass spectrometry studies published between the years 2017 and 2021, aiming at the determination of various compounds excreted in urine. The properties of the urine, including its diagnostical and analytical features and chemical composition, are also discussed including general protocols for the urine sample preparation. The mechanism of the electrophoretic separation and the instrumentation for capillary electrophoresis with mass spectrometry coupling is also included. This review shows the potential of the capillary electrophoresis with mass spectrometry technique for the analyses of different kinds of analytes in a complex biological matrix. The discussed applications are divided into two main groups (capillary electrophoresis with mass spectrometry for the determination of drugs and drugs of abuse in urine and capillary electrophoresis with mass spectrometry for the studies of urinary metabolome).

• Klíčová slova
• capillary electrophoresis, drugs, mass spectrometry, metabolome, urine analysis,
• MeSH
• elektroforéza kapilární metody   trendy   MeSH
• hmotnostní spektrometrie metody   trendy   MeSH
• léčivé přípravky moč   MeSH
• lidé MeSH
• metabolomika MeSH
• moč chemie   MeSH
• odhalování abúzu drog metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• léčivé přípravky MeSH

3-Quinuclidinyl benzilate (BZ) ranks among incapacitating military warfare agents. It acts as a competitive inhibitor on muscarinic receptors leading to non-lethal mental impairment. The present study aimed to investigate toxicokinetics of BZ in rats. Moreover, BZ can be exploited to produce a pharmacological model of Alzheimer's disease; thus, this paper focuses mainly on the BZ distribution to the brain. Wistar rats were administered i.p. with BZ (2 and 10 mg/kg). The BZ concentration was determined using LC-MS/MS in plasma, urine, bile, brain, kidney and liver. The sample preparation was based on a solid phase extraction (liquids) or protein precipitation (organ homogenates). The plasma concentration peaked at 3 min (204.5 ± 55.4 and 2185.5 ± 465.4 ng/ml). The maximal concentration in the brain was reached several minutes later. Plasma elimination half-life was 67.9 ± 3.4 in the 2 mg/kg group and 96.6 ± 27.9 in the 10 mg/kg group. BZ concentrations remained steady in the brain, with slow elimination (t1/2 506.9 ± 359.5 min). Agent BZ is excreted mainly via the urine. Steady BZ concentration in the brain could explain the previously published duration of the significant impairment in passive avoidance tasks in rats after an injection of BZ.

• Klíčová slova
• 3-quinuclidinyl benzilate, agent BZ, incapacitating agent, liquid chromatography-mass spectrometry, toxicokinetics,
• MeSH
• antagonisté muskarinových receptorů krev   metabolismus   toxicita   moč   MeSH
• chinuklidinylbenzilát krev   metabolismus   toxicita   moč   MeSH
• krysa rodu Rattus MeSH
• metabolom MeSH
• moč MeSH
• mozek metabolismus   MeSH
• potkani Wistar MeSH
• toxikokinetika MeSH
• žluč metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antagonisté muskarinových receptorů MeSH
• chinuklidinylbenzilát MeSH

BACKGROUND AND OBJECTIVES: Renal elimination of amikacin and other aminoglycosides is slowed down in sepsis-induced acute kidney injury increasing the risk of adverse effects. Since neutrophil gelatinase-associated lipocalin (NGAL) and aminoglycosides share the mechanisms for renal excretion, the predictive power of NGAL was examined towards the changes in amikacin pharmacokinetics during early endotoxemia in anesthetized Wistar rats. METHODS: Endogenous biomarkers of inflammation and acute kidney injury were assessed including NGAL in saline-injected controls and two groups of rats challenged with an intravenous injection of bacterial lipopolysaccharide (5 mg/kg)-a fluid-resuscitated group (LPS) and a fluid-resuscitated group infused intravenously with 8 μg/kg/h terlipressin (LPS-T). Sinistrin and amikacin were infused to measure glomerular filtration rate (GFR) and amikacin clearance (CLam). The investigations included blood gas analysis, chemistry and hematology tests and assessment of urine output, creatinine clearance (CLcr) and sinistrin clearance (CLsini). RESULTS: Within 3 h of injection, systemic and renal inflammatory responses were induced by lipopolysaccharide. Gene and protein expression of NGAL was increased in the kidneys and the concentrations of NGAL in the plasma (pNGAL) and urine rose 4- to 38-fold (P < 0.01). The decreases in CLam and the GFR markers (CLcr, CLsini) were proportional, reflecting the extent to which endotoxemia impaired the major elimination mechanism for the drug. Terlipressin attenuated lipopolysaccharide-induced renal dysfunction (urine output, CLcr, CLsini) and accelerated CLam. The pNGAL showed a strong association with the CLsini (rs = - 0.77, P < 0.0005). Concerning prediction of CLam, pNGAL was comparable to CLcr (mean error - 24%) and inferior to CLsini (mean error - 6.4%), while the measurement of NGAL in urine gave unsatisfactory results. CONCLUSIONS: During early endotoxemia in the rat, pNGAL has a moderate predictive ability towards CLam. Clinical studies should verify whether pNGAL can support individualized dosing of aminoglycosides to septic patients.

• MeSH
• akutní poškození ledvin krev   MeSH
• amikacin krev   metabolismus   farmakokinetika   MeSH
• biologické markery krev   MeSH
• cytokiny MeSH
• endotoxemie chemicky indukované   MeSH
• hodnoty glomerulární filtrace fyziologie   MeSH
• krysa rodu Rattus MeSH
• ledviny patofyziologie   MeSH
• lipokalin-2 krev   metabolismus   moč   MeSH
• lipopolysacharidy farmakologie   MeSH
• metabolická clearance MeSH
• moč MeSH
• modely u zvířat MeSH
• oligosacharidy farmakokinetika   MeSH
• potkani Wistar * MeSH
• prediktivní hodnota testů MeSH
• sepse farmakoterapie   metabolismus   MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amikacin MeSH
• biologické markery MeSH
• cytokiny MeSH
• lipokalin-2 MeSH
• lipopolysacharidy MeSH
• oligosacharidy MeSH
• sinistrin MeSH Prohlížeč

The cellular pool of purines is maintained by de novo purine synthesis (DNPS), recycling and degradation. Mutations in genes encoding DNPS enzymes cause their substrates to accumulate, which has detrimental effects on cellular division and organism development, potentially leading to neurological impairments. Unspecified neurological symptoms observed in many patients could not be elucidated even by modern techniques. It is presumable that some of these problems are induced by dysfunctions in DNPS enzymes. Therefore, we determined the concentrations of dephosphorylated DNPS intermediates by LC-MS/MS as markers of yet unpublished mutations in PFAS and PAICS genes connected with dysfunctions of carboxylase/phosphoribosylaminoimidazolesuccinocarboxamide synthase (PAICS) or phosphoribosylformylglycinamidine synthase (PFAS). We determined the criteria for normal values of metabolites and investigated 1,447 samples of urine and 365 dried blood spots of patients suffering from various forms of neurological impairment. We detected slightly elevated aminoimidazole riboside (AIr) concentrations in three urine samples and a highly elevated 5-formamidoimidazole-4-carboxamide riboside (FGAr) concentration in one urine sample. The accumulation of AIr or FGAr in body fluids can indicate PAICS or PFAS deficiency, respectively, which would be new disorders of DNPS caused by mutations in the appropriate genes. Measurement of DNPS intermediates in patients with neurological symptoms can uncover the cause of serious cellular and functional impairments that are otherwise inaccessible to detection. Further genetic and molecular analysis of these patients should establish the causal mutations for prenatal diagnosis, genetic consultation, and reinforce the DNPS pathway as a therapeutic target.

• MeSH
• lidé MeSH
• limita detekce MeSH
• metabolom MeSH
• metabolomika metody   MeSH
• moč MeSH
• mutace genetika   MeSH
• nemoci nervového systému genetika   metabolismus   MeSH
• puriny biosyntéza   krev   chemie   moč   MeSH
• test suché kapky krve MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• purine MeSH Prohlížeč
• puriny MeSH

The former perception of the urothelium as an impermeable barrier has been revised during the last decade, as increasing evidence of changes in urine composition during its passage of the urinary tract has been presented. Since differences in urothelial permeability between upper and lower urinary tract have been found, our aim is to demonstrate whether changes in urine composition occur during passage through the ureter. We studied consecutive urine samples from both renal pelvises in six pigs and compared them to samples from the bladder and distal ureter. We further sampled urine during storage in the bladder at a fixed volume. All samples were analysed by measuring osmolality and pH, along with the concentration of the following parameters: Na(+), K(+), Cl(-), creatinine, urea. Urine alkalinity increased significantly during passage of the ureter. Creatinine concentration, pH and K(+) increased significantly during the passage from pelvis to the bladder. All other parameters increased non-significantly during the passage to the bladder. The increase in concentration was more pronounced at low concentrations in the pelvis. During storage in the bladder, there was a significant increase in urea concentration. Changes in the composition of urine occur during its passage from the renal pelvis to the bladder and during storage in the bladder. Despite the brief transit time, significant changes in alkalinity were found already during passage through the ureter.

• MeSH
• časové faktory MeSH
• chloridy moč   MeSH
• draslík moč   MeSH
• koncentrace vodíkových iontů MeSH
• koncentrační schopnost ledvin * MeSH
• kreatinin moč   MeSH
• moč chemie   MeSH
• močovina moč   MeSH
• močový měchýř metabolismus   MeSH
• osmolární koncentrace MeSH
• sodík moč   MeSH
• Sus scrofa MeSH
• ureter metabolismus   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• chloridy MeSH
• draslík MeSH
• kreatinin MeSH
• močovina MeSH
• sodík MeSH

OBJECTIVE: Immunocytochemistry has attained a marginal role in urology so far. Combining the morphological and immunophenotypical changes of the urothelial cells retrieved from urine is a logical approach. The study aimed to analyse the diagnostic potential of immunocytological staining in the detection of high-grade and low-grade urothelial carcinoma. METHODS: Freshly voided urine was collected from 152 consecutive individuals, cytology classes were determined and cell blocks produced. A total of 77 patients were diagnosed with urothelial carcinoma and 75 patients had various benign urological conditions. Immunocytochemistry was performed using four antibodies: p53, MCM2, MCM5 and Ki-67. A diagnostic power to detect low grade and high-grade urothelial carcinoma was analysed for each antibody and their combinations with cytology. RESULTS: There were no significant differences between patients with low-grade tumours and control group. Antibodies p53 and Ki-67 slightly improved the sensitivity of urinary cytology while maintaining its specificity. The best negative predictive value was demonstrated in combinations of cytology and MCM5 (88.9%) and cytology, p53 and MCM5 (90.6%). In the diagnosis of high-grade tumours, all antibodies apart from MCM2 yielded better sensitivity and specificity than cytology alone (receiver operating characteristic curves: p53 = 0.853, MCM5 = 0.931, and Ki-67 = 0.895). Combined with cytology, the sensitivities went even higher for the cost of lower specificity. The best diagnostic performance was observed in the combination of MCM5 and Ki-67 (sensitivity = 96.2%; specificity = 80%). CONCLUSIONS: Immunocytochemistry with p53, MCM5 and Ki-67 antibodies can improve the diagnostic power of urinary cytology in the detection and follow-up of urinary bladder urothelial carcinoma.

• Klíčová slova
• diagnosis, follow-up, immunocytochemistry, urinary bladder, urinary cell block, urinary cytology, urothelial carcinoma,
• MeSH
• antigen Ki-67 imunologie   MeSH
• cytodiagnostika * MeSH
• imunohistochemie MeSH
• lidé MeSH
• MCM komplex, komponenta 2 imunologie   MeSH
• moč cytologie   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorový supresorový protein p53 imunologie   MeSH
• nádory močového měchýře diagnóza   patologie   MeSH
• proteiny buněčného cyklu imunologie   MeSH
• protilátky nádorové imunologie   MeSH
• ROC křivka MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• stupeň nádoru MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen Ki-67 MeSH
• MCM komplex, komponenta 2 MeSH
• MCM2 protein, human MeSH Prohlížeč
• MCM5 protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• nádorový supresorový protein p53 MeSH
• proteiny buněčného cyklu MeSH
• protilátky nádorové MeSH

BACKGROUNDS: The prevalence of toxoplasmosis is higher in schizophrenics than in the general population. It has been suggested that certain symptoms of schizophrenia, including changes in olfactory functions, are in fact symptoms of toxoplasmosis that can be easily detected in schizophrenics only due to the increased prevalence of toxoplasmosis in this population. Schizophrenics have impaired identification of odors and lower sensitivity of odor detection, however, no information about these parameters of non-schizophrenic Toxoplasma-infected subjects is available. METHODS: Here we searched for differences in olfactory functions between 62 infected and 61 noninfected non-schizophrenic subjects using the case-controls experimental design. RESULTS: The infected men scored better than the non-infected controls in the standard odor-identification test. The infected women rated all smells as more intensive while the infected men rated nearly all smells as less intensive. Infected women rated the pleasantness of the smell of the cat urine as higher than the non-infected women and the opposite was true for the men-in contrast, higher pleasantness of odor in infected men and lower in infected women were observed and described in the 2011 study. Toxoplasmosis, Rh, and toxoplasmosis-Rh interaction were not associated with the rated pleasantness of the smell of other stimuli. However, our sample contained only 17 Rh negative men and 30 Rh negative women. Therefore, all results concerning the main effects of Rh factor and the interaction with Rh factor must be considered only preliminary. CONCLUSIONS: Our results suggest that latent toxoplasmosis is associated with changes in the olfactory functions in humans; however, the observed changes differ from those observed in schizophrenics.

• MeSH
• dospělí MeSH
• kočky MeSH
• krevní skupiny - systém Rh-Hr metabolismus   MeSH
• lidé MeSH
• mladý dospělý MeSH
• moč MeSH
• odoranty MeSH
• schizofrenie metabolismus   parazitologie   patofyziologie   MeSH
• studie případů a kontrol MeSH
• Toxoplasma patogenita   MeSH
• toxoplazmóza metabolismus   patofyziologie   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• kočky MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• krevní skupiny - systém Rh-Hr MeSH

OBJECTIVE: The methodology of cell blocks (CBs) has long been an integrated part of cytology. However, there are very few data on CBs derived from urine. Their main disadvantage is a lack of cellularity, which limits their broader clinical applicability. Factors affecting cellular adequacy in urine remain unclear. We assessed the impact of basic clinical and cytopathological factors on the adequacy of cellularity in urinary CBs. METHODS: Freshly voided urine was collected from 401 consecutive individuals. Of these, 167 patients were diagnosed with urothelial carcinoma. The remaining 234 patients had various benign urological conditions. Papanicolaou classes were determined and CBs produced. Cellular adequacy was assigned to each CB (acellular, hypocellular, moderate cellularity, high cellularity), and moderately and highly cellular CBs were considered as adequate. Several factors were analysed to find any correlation with the adequacy of the cellularity. RESULTS: In univariate analysis, seven factors significantly correlated with the adequacy of the CBs. In the multivariate model, positive sediment (OR = 3.7), female sex (OR = 2.7), positive urinary cytology (OR = 2.6) and positive leucocyturia (OR = 2.1) were independent predictors of adequate cellularity. Positive predictive value and negative predictive value of the model were 65.0% and 77.7%, respectively. CONCLUSIONS: We determined four clinical and cytopathological factors which independently predict adequate cellularity in urinary CBs. Based on these results, several clinical situations have been proposed, in which the highest probability of adequate cellularity in urinary CBs can be achieved.

• Klíčová slova
• adequate cellularity, immunocytochemistry, urinary bladder, urinary cell block, urinary cytology, urothelial carcinoma,
• MeSH
• cytodiagnostika metody   MeSH
• karcinom z přechodných buněk diagnóza   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• moč cytologie   MeSH
• močový měchýř patologie   MeSH
• nádory močového měchýře diagnóza   patologie   MeSH
• následné studie MeSH
• senioři MeSH
• urologické nádory diagnóza   patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Simultaneous electromembrane extraction (EME) of six trace metal cations (Cu2+ , Zn2+ , Co2+ , Ni2+ , Pb2+ , Cd2+ ) from saline samples was investigated. CE with capacitively coupled contactless conductivity detection (C4 D) was used to determine the metals in acceptor solutions due to its excellent compatibility with the minute volumes of acceptor solutions. Bis(2-ethylhexyl)phosphate (DEHPA) was selected as a suitable nonselective modifier for EME transport of target metal cations. Both, the individual effect of each major inorganic cation (Na+ , K+ , Ca2+ , Mg2+ ) and their synergistic effect on EME of the trace metal cations were evaluated. In both cases, a decrease in extraction efficiency was observed when major inorganic cations were present in the sample. This effect was more significant for Ca2+ and Mg2+ . The system was optimized for simultaneous extractions of the six target metals from saline samples (50 mM Na+ , 5 mM Mg2+ , 1 mM K+ , and 1 mM Ca2+ ) and following EME conditions were applied. Organic phase consisted of 1-nonanol containing 1% (v/v) DEHPA, acceptor solution was 1 M acetic acid (HAc) and sample pH was adjusted to 5. Sample was stirred at 750 rpm and EMEs were carried out at extraction potential of 10 V for 20 min. The method presented a repeatability between 8 and 21.8% (n = 5), good linearity in 0.5-10 μM concentration range (R2 = 0.987-0.999) and LOD better than 2.6 nM. Applicability of the EME-CE-C4 D method to the analyses of metal cations in drinking water, seawater, and urine samples was also demonstrated.

• Klíčová slova
• CE, DEHPA, Electromembrane extraction, Major inorganic cations, Metal cations, Saline samples,
• MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární metody   normy   MeSH
• moč chemie   MeSH
• mořská voda chemie   MeSH
• pitná voda chemie   MeSH
• solný roztok MeSH
• stopové prvky analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• pitná voda MeSH
• solný roztok MeSH
• stopové prvky MeSH