Human peripheral blood T cells or continuous T cell lines were treated with phorbol myristate (PMA), a direct protein kinase C activator. The effect of isoniazid on PMA-treated cells was investigated. It was found that low doses of isoniazid augmented proliferation of T cells treated with PMA or stimulated with PMA and phytohemagglutinin. Similar results were shown in the Jurkat cell line. There was no influence of isoniazid upon proliferation of PMA-treated HT-2 cells, interleukin 2-dependent cell line. The obtained results suggest that, at least one target mechanism of isoniazid is located in T cell activation pathway after protein kinase C activation.
- MeSH
- adjuvancia imunologická farmakologie MeSH
- aktivace lymfocytů účinky léků MeSH
- buněčné linie MeSH
- fytohemaglutininy farmakologie MeSH
- isoniazid farmakologie MeSH
- lidé MeSH
- tetradekanoylforbolacetát farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- adjuvancia imunologická MeSH
- fytohemaglutininy MeSH
- isoniazid MeSH
- tetradekanoylforbolacetát MeSH
Hydrogen peroxide production was measured in non-elicited rat peritoneal macrophages using luminol-dependent chemiluminescence (LDCL). Isolated cells were activated by a chemotactic peptide (FMLP) or by a phorbol ester (PMA) or by the combination of both. A hundred-fold higher LDCL intensity was achieved with PMA relative to FMLP. However, when FMLP was added subsequently to PMA it produced approximately the same response as did PMA. These measurements were carried out with cells isolated from controls and from animals exposed to normobaric hypoxia (10% O2) for 3 hours, 3 days, or 21 days. Hypoxia had a dual effect. Acutely (within 3 hours) it attenuated the production of hydrogen peroxide triggered by PMA, whilst during longer exposure (3 or 21 days) it increased the response induced by FMLP. Hypoxia can thus modulate the capacity of respiratory burst in peritoneal macrophages.
- MeSH
- hypoxie metabolismus MeSH
- krysa rodu Rattus MeSH
- luminiscenční měření MeSH
- luminol MeSH
- N-formylmethionin-leucyl-fenylalanin farmakologie MeSH
- peritoneální makrofágy účinky léků metabolismus MeSH
- peroxid vodíku metabolismus MeSH
- potkani Wistar MeSH
- spotřeba kyslíku MeSH
- tetradekanoylforbolacetát farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- luminol MeSH
- N-formylmethionin-leucyl-fenylalanin MeSH
- peroxid vodíku MeSH
- tetradekanoylforbolacetát MeSH
Mycobacterium avium subsp. paratuberculosis (MAP) is a pathogenic bacterium causing the paratuberculosis, chronic and infectious disease common particularly in wild and domestic ruminants. Currently, culture techniques to detect viable MAP are still used most commonly, although these require a long incubation period. Consequently, a faster molecular method for assessing MAP cell viability based on cell membrane integrity was introduced consisting of sample treatment with the intercalation dye propidium monoazide (PMA) followed by quantitative PCR (qPCR). However, the PMA-qPCR assay is complicated by demanding procedures involving work in a darkroom and on ice. In this study, we therefore optimized a viability assay combining sample treatment with palladium (Pd) compounds as an alternative viability marker to PMA, which does not require such laborious procedures, with subsequent qPCR. The optimized Pd-qPCR conditions consisting of 90 min exposure to 30 µM bis(benzonitrile)dichloropalladium(II) or 30 µM palladium(II)acetate at 5 °C and using ultrapure water as a resuspension medium resulted in differences in quantification cycle (Cq) values between treated live and dead MAP cells of 8.5 and 7.9, respectively, corresponding to approximately 2.5 log units. In addition, Pd-qPCR proved to be superior to PMA-qPCR in distinguishing between live and dead MAP cells. The Pd-qPCR viability assay thus has the potential to replace time-consuming culture methods and demanding PMA-qPCR in the detection and quantification of viable MAP cells with possible application in food, feed, clinical and environmental samples.
- MeSH
- azidy farmakologie MeSH
- biotest MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- mikrobiální viabilita MeSH
- Mycobacterium avium subsp. paratuberculosis * genetika MeSH
- palladium farmakologie MeSH
- paratuberkulóza * mikrobiologie MeSH
- propidium farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- azidy MeSH
- palladium MeSH
- propidium MeSH
We have studied in vitro alveolar macrophages (AMs) obtained by tracheobronchial lavage from rats exposed to subacute (3 hours and 3 days) and chronic (3 weeks) hypoxia (FiO2 = 0.1) and from rats recovering from chronic hypoxia. Hydrogen peroxide production by AMs was measured by luminol-dependent chemiluminescence after AMs adhered to the walls of the measuring cuvette, after stimulation with phorbol-myristate-acetate (PMA), and when N-formyl-methionyl-leucyl-phenylanine (FMLP) was added subsequently to the cells which had been previously stimulated by adherence or PMA. H2O2 production after cell adherence and adherence combined with FMLP stimulation did not differ between the groups. The increase of H2O2 production after adding PMA, and FMLP in addition to PMA was significantly higher in AMs from rats exposed to hypoxia for 3 days than in the controls. Other experimental groups did not differ from their controls. It is concluded that 3 days' hypoxia primes AMs for enhanced production of H2O2 upon stimulation. The mechanism is probably at the level of synthesis of proteins involved in H2O2 production, or the shift to a more reactive phenotype of alveolar macrophages subpopulations.
- MeSH
- alveolární makrofágy metabolismus MeSH
- buněčná adheze fyziologie MeSH
- hypoxie metabolismus MeSH
- krysa rodu Rattus MeSH
- luminiscenční měření MeSH
- N-formylmethionin-leucyl-fenylalanin farmakologie MeSH
- peroxid vodíku metabolismus MeSH
- potkani Wistar MeSH
- reaktivní formy kyslíku metabolismus MeSH
- tetradekanoylforbolacetát farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- N-formylmethionin-leucyl-fenylalanin MeSH
- peroxid vodíku MeSH
- reaktivní formy kyslíku MeSH
- tetradekanoylforbolacetát MeSH
BACKGROUND: The aim of our study was to compare serum concentrations of adiponectin, leptin and other selected parameters in female patients with restrictive subtype of anorexia nervosa (n=15), (RMA), binge/purge subtype of anorexia nervosa (n=11) (PMA) with age-matched healthy females, (C, n=14). METHODS AND RESULTS: RMA patients had the most severely decreased body mass index (BMI) and serum leptin levels of the three groups studied. These parameters were also significantly lower in PMA relative to C group. (BMI: RMA 14.61 +/- 0.49 kg/m2, PMA 17.30 +/- 0.25 kg/m2, C 23.21 +/- 0.96 kg/m2; leptin: RMA 1.39 +/- 0.31 ng/ml, PMA 3.72 +/- 0.77 ng/ml, C 9.17 +/- 1.53 ng/ml). In contrast, serum adiponectin levels were markedly increased in RMA patients (57.28 +/- 4.86 ug/ml) relative to other groups (PMA 40,25 +/- 2.18 microg/ml, K 26.84 +/- 2.40 microg/ml). Serum leptin levels positively correlated with BMI in all groups studied (r = 0.56, p = 0.002), while the inverse relationship was found for adiponectin levels and BMI (r = -0.72, p = 0.000003). The hormonal concentrations were measured by commercially available RIA and ELISA kits. CONCLUSIONS: The most significant changes of serum adiponectin and leptin levels were found in the RMA group with most severely decreased BMI and body fat content relative to rest of the groups. Possible role of increased adiponectin levels in the etiopathogenesis and/or metabolic changes in patients with anorexia nervosa is under the scope of our current investigations.
- MeSH
- adiponektin MeSH
- bulimia krev MeSH
- index tělesné hmotnosti MeSH
- leptin krev MeSH
- leptinové receptory MeSH
- lidé MeSH
- mentální anorexie krev MeSH
- mezibuněčné signální peptidy a proteiny krev MeSH
- receptory buněčného povrchu krev MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- adiponektin MeSH
- leptin MeSH
- leptinové receptory MeSH
- mezibuněčné signální peptidy a proteiny MeSH
- receptory buněčného povrchu MeSH
In this study we examined whether microtubules and heat shock protein 90 (Hsp90) are involved in phorbol myristate acetate (PMA) and N-formyl-Met-Leu-Phe (fMLP)-induced oxidative burst in DMSO-differentiated HL-60 cells. Our results showed that microtubule interfering agents, paclitaxel (1-5 microM), colchicine (1-100 microM), nocodazole (1-20 microM), and vincristine (1-50 microM), did not affect either PMA or fMLP-induced oxidative burst. In contrast, radicicol, an inhibitor of Hsp90, inhibited fMLP-induced oxidative burst in time and concentration-dependent manner where IC50 value for 30 min pre-incubation was 16.5 +/- 3.5 microM radicicol. We conclude that both PMA and fMLP-induced oxidative burst in DMSO-differentiated HL-60 cells is microtubule-independent while the latter requires Hsp90 activity.
- MeSH
- buněčná diferenciace MeSH
- dimethylsulfoxid farmakologie MeSH
- HL-60 buňky MeSH
- kolchicin farmakologie MeSH
- lidé MeSH
- mikrotubuly účinky léků fyziologie MeSH
- N-formylmethionin-leucyl-fenylalanin farmakologie MeSH
- neutrofily metabolismus MeSH
- nokodazol farmakologie MeSH
- paclitaxel farmakologie MeSH
- proteiny tepelného šoku HSP90 metabolismus MeSH
- respirační vzplanutí účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dimethylsulfoxid MeSH
- kolchicin MeSH
- N-formylmethionin-leucyl-fenylalanin MeSH
- nokodazol MeSH
- paclitaxel MeSH
- proteiny tepelného šoku HSP90 MeSH
Activated polymorphonuclear leukocytes (PMNL) might be an important source of host tissue damage. PMNL may be primed by various inflammatory mediators for an increased production of reactive oxygen species (ROS). The short-term priming effects of two cytokines, interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), on rat PMNL in whole unfractionated blood were investigated in the present study. Incubation of PMNL with cytokine alone did not induce the detectable ROS production without addition of a second stimulus-phorbol myristate acetate (PMA) or opsonized zymosan particles (OZP). One-hour incubation with TNF-alpha, or the mixture of TNF-alpha + IFN-gamma resulted in a dose-dependent enhancement of ROS production in response to PMA. Moreover, the incubation with both IFN-gamma and TNF-alpha caused a significantly higher increase of ROS production than with TNF-alpha alone. However, no priming effects of IFN-gamma on the PMA-induced ROS production were observed. Finally, none of the cytokines enhanced the total production of ROS upon the stimulation with OZP, although changes in the time course of ROS production were observed. Thus, different signal transduction pathways seem to be involved in PMA- and OZP-induced respiratory burst. Alternatively, partial activation and assembling of NADPH oxidase during cytokine treatment might explain the differences observed between PMA- and OZP-induced ROS production. The results of the present study suggest that IFN-gamma may enhance the priming effects of TNF-alpha. This finding may have implications for understanding the mechanisms by which both cytokines may contribute to PMNL-mediated tissue injury during various clinical conditions, as well as to host defense against invading pathogens.
- MeSH
- interferon gama farmakologie MeSH
- krev účinky léků MeSH
- krysa rodu Rattus MeSH
- luminiscenční měření MeSH
- neutrofily účinky léků MeSH
- potkani Wistar MeSH
- reaktivní formy kyslíku metabolismus MeSH
- rekombinantní proteiny farmakologie MeSH
- respirační vzplanutí účinky léků MeSH
- tetradekanoylforbolacetát farmakologie MeSH
- TNF-alfa genetika farmakologie MeSH
- zvířata MeSH
- zymosan farmakologie MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- interferon gama MeSH
- reaktivní formy kyslíku MeSH
- rekombinantní proteiny MeSH
- tetradekanoylforbolacetát MeSH
- TNF-alfa MeSH
- zymosan MeSH
The size and structural changes of nanoparticles formed after the addition of poly(2-vinylpyridine), PVP, to block copolymer micelles of polystyrene-block-poly(methacrylic acid), PS-PMA, were studied by light scattering and atomic force microscopy. Due to the strong hydrogen bonding between PVP and PMA segments, complex structures based on the core/shell micelles form in mixed selective solvents. As proven by a combination of light scattering and atomic force microscopy, individual PS-PMA micelles are "glued" together by PVP chains. The dialysis against solvents with a high content of water results in transient increase in polydispersity and turbidity of originally clear solutions. However, the precipitated polymer material dissolves in basic buffers and stable soluble nanoparticles reform in aqueous media. The behavior of their solutions was studied in a broad pH range by light scattering, atomic force microscopy and capillary zone electrophoresis.
- Publikační typ
- časopisecké články MeSH
UNLABELLED: The utility of decision tree machine learning in exploring the interactions among the SpO2 target range, neonatal maturity, and oxemic-risk is demonstrated. METHODS: This observational study used 3 years of paired age-SpO2-PaO2 data from a neonatal ICU. The CHAID decision tree method was used to explore the interaction of postmenstrual age (PMA) on the risk of extreme arterial oxygen levels at six different potential SpO2 target ranges (88-92%, 89-93%, 90-94%, 91-95%, 92-96% and 93-97%). Risk was calculated using a severity-weighted average of arterial oxygen outside the normal range for neonates (50-80 mmHg). RESULTS: In total, 7500 paired data points within the potential target range envelope were analyzed. The two lowest target ranges were associated with the highest risk, and the ranges of 91-95% and 92-96% were associated with the lowest risk. There were shifts in the risk associated with PMA. All the target ranges showed the lowest risk at ≥42 weeks PMA. The lowest risk for preterm infants was within a target range of 92-96% with a PMA of ≤34 weeks. CONCLUSIONS: This study demonstrates the utility of decision tree analytics. These results suggest that SpO2 target ranges that are different from typical range might reduce morbidity and mortality. Further research, including prospective randomized trials, is warranted.
- Klíčová slova
- decision tree classification, hyperoxemic-risk, hypoxemic-risk, machine learning, neonatal, oxygen saturation targeting,
- Publikační typ
- časopisecké články MeSH
Activation of neutrophils induces generation of reactive oxygen species and release of granule enzymes, which not only participate in the bactericidal mechanisms of these cells, but also in possible tissue damage. We studied the effect of carvedilol (CARV) [0.1-100 micromol/l], an antihypertensive and cardiovascular drug with antioxidative properties, on superoxide generation (SO) and myeloperoxidase (MPO) release from isolated human neutrophils stimulated with fMLP, a specific receptor activator, or with PMA, a receptor bypassing stimulus. Unstimulated cells showed neither SO formation nor MPO release after preincubation with drug. CARV decreased fMLP and PMA stimulated MPO release and SO generation dose dependently. The inhibitory effect of CARV may attributed to non-specific action since its effect was not influenced by the type of stimulation. It might inhibit SO generation as well as MPO release either by membrane-operating stimulus (fMLP) or membrane bypassing activator (PMA).
- MeSH
- aktivace neutrofilů * MeSH
- antihypertenziva farmakologie MeSH
- antioxidancia farmakologie MeSH
- karbazoly farmakologie MeSH
- karvedilol MeSH
- lidé MeSH
- neutrofily metabolismus MeSH
- peroxidasa metabolismus MeSH
- propanolaminy farmakologie MeSH
- superoxidy metabolismus MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antihypertenziva MeSH
- antioxidancia MeSH
- karbazoly MeSH
- karvedilol MeSH
- peroxidasa MeSH
- propanolaminy MeSH
- superoxidy MeSH
