• MeSH
• alveolární makrofágy enzymologie   imunologie   účinky léků   MeSH
• chronická obstrukční plicní nemoc * farmakoterapie   imunologie   patofyziologie   MeSH
• dendritické buňky imunologie   účinky léků   MeSH
• eozinofily imunologie   účinky léků   MeSH
• lidé MeSH
• mediátory zánětu diagnostické užití   imunologie   MeSH
• neutrofily enzymologie   imunologie   účinky léků   MeSH
• zánět * diagnóza   imunologie   MeSH
• Check Tag
• lidé MeSH

To specify the site of action of the synthetic coumarin derivatives 7-hydroxy-3-(4'-hydroxyphenyl) coumarin (HHC) and 7-hydroxy-3-(4'-hydroxyphenyl) dihydrocoumarin (HHDC), we evaluated their effects on extra- and intracellular reactive oxygen species (ROS) formation in phorbol-myristate-13-acetate (PMA) stimulated human neutrophils. We studied also the effects of HHC and HHDC on possible molecular mechanisms which participate in the activation of NADPH oxidase, that is, on PKC activity, on phosphorylation of some PKC isoforms (α, βII, and δ), and on phosphorylation of the NADPH oxidase subunit p40(phox). Without affecting cytotoxicity, both coumarines tested were effective inhibitors/scavengers of ROS produced by neutrophils on extracellular level. HHC markedly diminished oxidant production and also, intracellularly, decreased PKC activity and partly phosphorylation of PKCα, βII. On the other hand, we did not observe any effect of coumarin derivatives on phosphorylation of PKC δ and on phosphorylation of the NADPH oxidase subunit p40(phox), which were suggested to be involved in the PMA-dependent intracellular activation process. In agreement with our previous findings, we assume that the different molecular structures of HHC and HHDC with their different physicochemical and free radical scavenging characteristics are responsible for their diverse effects on the parameters tested.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• aktivace neutrofilů účinky léků   MeSH
• bezbuněčný systém MeSH
• buněčná smrt účinky léků   MeSH
• dospělí MeSH
• extracelulární prostor účinky léků   metabolismus   MeSH
• fosfoproteiny metabolismus   MeSH
• fosforylace účinky léků   MeSH
• inhibiční koncentrace 50 MeSH
• intracelulární prostor účinky léků   metabolismus   MeSH
• izoenzymy metabolismus   MeSH
• kinetika MeSH
• kumariny chemie   farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• luminiscenční měření MeSH
• mladý dospělý MeSH
• neutrofily cytologie   účinky léků   enzymologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• tetradekanoylforbolacetát farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

PURPOSE OF THE STUDY: Leukocyte esterase is an enzyme in neutrophils from which it is released into exudate; its detection by colorimetric test strips indicates the presence of neutrophils. This is a rapid method to find whether exudate is of infectious or non-infectious aetiology. The aim of the study was to determine the sensitivity and specificity of leukocyte esterase testing with use of AUTION Sticks (Arkray) for examination of exudates obtained in inflammatory diseases of the skeletal system. MATERIAL AND METHODS: Exudates associated with skeletal system diseases were collected from 45 patients in the period from July 1st to December 31 st , 2012. Aspirates obtained under sterile conditions were examined for leukocyte esterase; cytological and microbiological examinations were also carried out. For the detection of leukocyte esterase, a drop of aspirate was placed on the reagent zone of a test strip and the resulting colour reaction was read after 90 minutes. Changes in colour were compared with a reference strip provided by the manufacturer. The results were assessed on a five-shade scale as follows: 0, no colour change; 1 to 4, gradual change from light pink to deep purple. The results were compared with those of cytological and microbiological examinations. Shade 4 on the strip corresponded to a positive cytological finding of bacterial infection, and shades 3 and 4 correlated with a positive microbial finding. The sensitivity and specificity of leukocyte esterase testing were statistically evaluated for both comparisons. RESULTS: Based on the results of cytological and microbiological examinations, an infectious aetiology of exudate was diagnosed in 21 (44.4%) and non-infectious aetiology in 24 (63.6%) patients. With leukocyte esterase reagent strips when shade 4 was taken as a positive result, the sensitivity and specificity of examination was assessed as 0.6190 and 0.9583, respectively. When taking both shade 3 and shade 4 for a positive result, sensitivity and specificity were 0.8571 and 0.8750, respectively. Shades 0 and 1 corresponded to the number of leukocytes in exudate that was no higher than 2 x 10⁹/ml. DISCUSSION: The detection of leukocyte esterase is a quick and easy examination. It is useful for readily excluding or confirming an infectious aetiology of exudate and can, to some extent, substitute a cytological examination. It can also help to make a quick decision whether one- or two-stage joint reimplantation should be performed and thus eliminate the need of intra-operative histological examination of frozen tissue samples. A drawback of the method was that exudate samples contaminated with blood interfered with an assessment of colour shades. However, this can be avoided by centrifugation of the sample and use of a supernatant free from erythrocytes. CONCLUSIONS: Diagnosing infectious aetiology of joint exudate or exudate from an abscess using leukocyte esterase reagent strips appears, according to our results, to be a promising, semi-quantitative method with high specificity and sensitivity which is rapid, simple and affordable. It can be useful particularly in out-patient institutions for a quick diagnosis of arthritis; intraoperatively, it can serve as an additional method to other exudate examinations.

• MeSH
• absces diagnóza   mikrobiologie   MeSH
• artritida diagnóza   mikrobiologie   MeSH
• bakteriální infekce diagnóza   MeSH
• cytodiagnostika MeSH
• exsudáty a transsudáty enzymologie   MeSH
• karboxylesterhydrolasy analýza   MeSH
• lidé MeSH
• nemoci kostí diagnóza   mikrobiologie   MeSH
• neutrofily enzymologie   MeSH
• reagenční papírky * MeSH
• senzitivita a specificita MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• srovnávací studie MeSH

• Klíčová slova
• pterostilben,
• MeSH
• aktivace neutrofilů účinky léků   MeSH
• antioxidancia MeSH
• lidé MeSH
• neutrofily * enzymologie   MeSH
• oxidační stres imunologie   účinky léků   MeSH
• peroxidasa MeSH
• proteinkinasa C MeSH
• Pterocarpus MeSH
• reaktivní formy kyslíku škodlivé účinky   MeSH
• stilbeny * farmakologie   chemie   MeSH
• superoxidy * škodlivé účinky   MeSH
• techniky in vitro MeSH
• výzkum MeSH
• zánět prevence a kontrola   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Matrix-metaloproteinázy (MMP) a jejich tkáňové inhibitory TIMP jsou zapojeny do patogeneze chronické obstrukční plicní nemoci (CHOPN) a bronchiálního astmatu. Obě onemocnění se liší typem zánětu, imunologickými mechanismy a jsou podmíněny různými buňkami a mediátory. Zatím existuje málo informací o rozdílech v expresních profilech jednotlivých MMP a TIMP stejně jako v zastoupení jednotlivých buněčných subpopulací bronchoalveolárních buňek pacientů s CHOPN a bronchiálním astmatem. Zkoumali jsme proto buněčné profily a genovou expresi MMP-2, MMP-9 a TIMP-1 v bronchoalveolárních buňkách získaných z bronchoalveolární laváže (BAL) u pacientů s CHOPN (n=32), s perzistujícím bronchiálním astmatem (n=24) a u kontrolních subjektů (n=16) pomocí kvantitativní RT-PCR. Srovnáním zastoupení jednotlivých buněčných populací v bronchoalveolární tekutině (BALTe) jsme potvrdili výraznou neutrofilii u pacientů ve všech stádiích CHOPN a výraznou eosinofilii u pacientů s astmatem. U pacientů s CHOPN jsme zjistili signifikantně zvýšenou expresi mRNA MMP-2 (relativní exprese ± SD: 0,08 ± 0,06 vs. 0,03 ± 0,02, p = 0,007), MMP-9 (0,12 ± 0,14 vs. 0,02 ± 0,02, p = 0,003) a TIMP-1 (3,08 ± 3,79 vs. 0,97 ± 0,66, p = 0,004) ve srovnání s kontrolní skupinou. U pacientů s astmatem jsme rovněž zjistili signifikantně zvýšenou expresi mRNA MMP-2 (0,10 ± 0,10 vs. 0,03 ± 0,02, p = 0,002), MMP-9 (0,16 ± 0,15 vs. 0,02 ± 0,02, p = 0,001) a TIMP-1 (2,27 ± 2,64 vs. 0,97 ± 0,66, p = 0,003) ve srovnání s kontrolní skupinou, přitom rozdíly mezi pacienty s CHOPN a astmatem nebyly statisticky významné. Rozdíly v expresi MMP-2, MMP-9 a TIMP-1 mezi 1., 2. a 3. stádiem CHOPN nedosáhly statistické významnosti. Ve skupině pacientů s 2. stadiem CHOPN byla proti kontrolní skupině signifikantně zvýšená exprese MMP-2 (p=0,002), MMP-9 (p=0,008) a TIMP-1 (p=0,008), ve skupině se 3.stádiem CHOPN byla signifikantně zvýšená exprese TIMP-1 (p=0,018) proti kontrolní skupině, ostatní rozdíly nebyly statisticky významné. Výsledky naší práce ukazují, že exprese mRNA MMP-2, MMP-9 a TIMP-1 v bronchoalveolárních buňkách jsou zvýšeny u pacientů s CHOPN i bronchiálním astmatem. Dále jsme prokázali zvýšenou expresi mRNA MMP-9 a TIMP-1 u pacientů v těžších stadiích CHOPN.

Matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) are involved in the pathogenesis of chronic obstructive pulmonary disease (COPD) and bronchial asthma. The two diseases differ by the type of inflammation and immunological mechanisms and are conditioned by different cells and mediators. Little is known about differences in expression profiles of individual MMP and TIMP as well as in representation of individual cell subpopulations of bronchoalveolar cells in patients with COPD and bronchial asthma. Therefore, cell profiles and gene expression of MMP-2, MMP-9 and TIMP-1 in bronchoalveolar cells obtained by bronchoalveolar lavage (BAL) in patients with COPD (n = 32) or persistent bronchial asthma (n = 24) and in controls (n = 16) were tested by quantitative RT-PCR. The proportion of individual cell populations in the bronchoalveolar lavage fluid (BALE) was compared. Marked neutrophilia in patients with all stages of COPD and marked eosinophilia in asthma patients was conflrmed. Patients with COPD were found to have significantly increased expression of mRNA for MMP-2 (relative expression ± SD: 0.08 ± 0.06 vs. 0.03 ± 0.02, p = 0.007), MMP-9 (0.12 ± 0.14 vs. 0.02 ± 0.02, p = 0.003) and TlMP-1 (3.08 ± 3.79 vs. 0.97 ± 0.66, p = 0.004) as compared with controls. Asthma patients were also found to have significantly increased expression of mRNA for MMP-2 (0.10 ± 0.10 vs. 0.03 ± 0.02, p = 0.002), MMP-9 (0.16 ± 0.15 VS. 0.02 ± 0.02, p = 0.001) and TIMP-1 (2.27 ± 2.64 vs. 0.97 ± 0.66, p = 0.003) as compared with controls, with statistically insignificant differences between COPD and asthma patients. Differences in MMP-2, MMP-9 and TIMP-1 expression between COPD stages 1, 2 and 3 were not statistically significant. Patients with stage 2 COPD had significantly increased expression of MMP-2 (p = 0.002), MMP-9 (p = 0.008) and TIMP-1 (p = 0.008) and stage 3 COPD patients had significantly increased expression of TIMP-1 (p = 0.018), as compared with controls. The other differences were not statistically significant. The results suggest that expression of mRNA for MMP-2, MMP-9 and TIMP-1 in bronchoalveolar cells is increased in patients with both COPD and bronchial asthma. Additionally, increased expression of mRNA for MMP-9 and TlMP-1 was found in patients with more severe COPD stages.

• Klíčová slova
• tkáňové inhibitory, astma bronchiale,
• MeSH
• bronchiální astma enzymologie   imunologie   patofyziologie   MeSH
• bronchoalveolární lavážní tekutina cytologie   chemie   imunologie   MeSH
• chronická obstrukční plicní nemoc enzymologie   imunologie   patofyziologie   MeSH
• eozinofily cytologie   enzymologie   MeSH
• financování organizované MeSH
• lidé MeSH
• metaloproteinasy secernované do matrix diagnostické užití   MeSH
• neutrofily cytologie   enzymologie   MeSH
• pilotní projekty MeSH
• tkáňové inhibitory metaloproteinas diagnostické užití   MeSH
• Check Tag
• lidé MeSH

The quaternary benzo[c]phenanthridine alkaloid chelerythrine is widely used as an inhibitor of protein kinase C (PKC). However, in biological systems chelerythrine interacts with an array of proteins. In this study, we examined the effects of chelerythrine and sanguinarine on conventional PKCs (cPKCs) and PKC upstream kinase, phosphoinositide-dependent protein kinase 1 (PDK1), under complete inhibition conditions of PKC-dependent oxidative burst. In neutrophil-like HL-60 cells, sanguinarine and chelerythrine inhibited N-formyl-Met-Leu-Phe, phorbol 12-myristate 13-acetate (PMA)-, and A23187-induced oxidative burst with IC(50) values not exceeding 4.6 micromol/L, but the inhibition of PMA-stimulated cPKC activity in intact cells required at least fivefold higher alkaloid concentrations. At concentrations below 10 micromol/L, sanguinarine and chelerythrine prevented phosphorylation of approximately 80 kDa protein and sequestered approximately 60 kDa phosphoprotein in cytosol. Moreover, neither sanguinarine nor chelerythrine impaired PMA-stimulated translocation of autophosphorylated PKCalpha/betaII isoenzymes, but both alkaloids induced dephosphorylation of the turn motif in PKCalpha/betaII. The dephosphorylation did not occur in unstimulated cells and it was not accompanied by PKC degradation. Furthermore, cell treatment with sanguinarine or chelerythrine resulted in phosphorylation of approximately 70 kDa protein by PDK1. We conclude that PKC-dependent cellular events are affected by chelerythrine primarily by multiple protein interactions rather than by inhibition of PKC activity.

• MeSH
• alkaloidy farmakologie   chemie   MeSH
• benzofenantridiny farmakologie   chemie   MeSH
• bezbuněčný systém MeSH
• buněčná smrt účinky léků   MeSH
• fosforylace účinky léků   MeSH
• HL-60 buňky MeSH
• isochinoliny farmakologie   chemie   MeSH
• izoenzymy metabolismus   MeSH
• lidé MeSH
• NADPH-oxidasy metabolismus   MeSH
• neutrofily cytologie   enzymologie   účinky léků   MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• proteinkinasa C metabolismus   MeSH
• respirační vzplanutí účinky léků   MeSH
• substrátová specifita účinky léků   MeSH
• transport proteinů účinky léků   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

OBJECTIVE: Progelatinase B/proMMP-9 has recently been identified as an indicator of pleural inflammation, presumably originating from granulocytes. The aim of this study was to verify the origin of progelatinase B by simultaneous estimation of specific markers of neutrophil recruitment and activation in pleural effusions following induced pleurisy and pleural injury. MATERIAL AND METHODS: Sixty-three samples of pleural fluid from patients undergoing therapeutic talc pleurodesis (n = 8) and explorative thoracoscopy (n = 3) collected before and at different time intervals after the intervention were analyzed for progelatinase B and neutrophil gelatinase-associated lipocalin (NGAL)-gelatinase complex by substrate electrophoresis, for myeloperoxidase (MPO) and interleukin-8 (IL-8) by immunoadsorbent sandwich assay, as well as for leukocyte count, C-reactive protein (CRP) and total protein (TP). RESULTS: A significant increase in free and NGAL-complexed progelatinase B, MPO and IL-8 was recorded within 48 h following treatment in all subjects. Progelatinase B was strongly correlated with NGAL-gelatinase complex (r = 0.88, p = 0.001), MPO (r = 0.81, p = 0.001), neutrophil count (r = 0.75, p = 0.01) and IL-8 (r = 0.71, p = 0.001), but not with CRP and TP. CONCLUSIONS: The results support the neutrophil origin of the proenzyme, which confirms progelatinase B as an indicator of a local inflammatory reaction. Quantifying the inflammatory reaction may be helpful in the evaluation of both the technical variants of therapeutic pleurodesis and finer discrimination of paraneoplastic effusions.

• MeSH
• biologické markery metabolismus   MeSH
• degranulace buněk fyziologie   MeSH
• financování organizované MeSH
• interleukin-8 metabolismus   MeSH
• kolagenasy biosyntéza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipokaliny MeSH
• matrixová metaloproteinasa 9 MeSH
• metaloendopeptidasy biosyntéza   MeSH
• neutrofily enzymologie   fyziologie   patologie   MeSH
• peroxidasa metabolismus   MeSH
• pleura enzymologie   patologie   zranění   MeSH
• pleurální výpotek enzymologie   patologie   MeSH
• pleuritida enzymologie   patologie   MeSH
• prekurzory enzymů biosyntéza   MeSH
• proteiny akutní fáze metabolismus   MeSH
• protoonkogenní proteiny metabolismus   MeSH
• senioři MeSH
• želatinasy biosyntéza   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH

• MeSH
• antioxidancia metabolismus   MeSH
• diabetes mellitus 2. typu enzymologie   krev   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• lineární modely MeSH
• neutrofily enzymologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

• MeSH
• glukuronidasa analýza   MeSH
• lidé MeSH
• lyzozomy enzymologie   účinky léků   MeSH
• neutrofily enzymologie   patologie   účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• techniky in vitro MeSH

• MeSH
• dieta MeSH
• dítě MeSH
• glykogen MeSH
• lidé MeSH
• neutrofily enzymologie   patofyziologie   MeSH
• neutropenie enzymologie   imunologie   MeSH
• předškolní dítě MeSH
• vrozené poruchy metabolismu sacharidů dietoterapie   imunologie   komplikace   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH