Průduškové astma (astma) je jedním z nejčastějších chronických zánětlivých onemocnění v české populaci. Obecně se řadí mezi onemocnění s nedostatečnou nebo neadekvátně regulovanou systémovou zánětlivou aktivitou, projevující se chronickými zánětlivými změnami v cestách dýchacího systému. V léčbě je užívána komplexní léčebná strategie, stupňovitě vedená, jejímž farmakologickým základem je aplikace imunomodulačních léčiv. Lékem volby, ale i nejčastěji aplikovaným imunomodulačním léčivem, jsou inhalační glukokortikoidy. Spíše ojediněle je u jedinců s astmatem prokázána glukokortikoidní rezistence, která je definována termínem kortikorezistentní astma. Jedinci s kortikorezistentním astmatem nejen že neprofitují z terapie glukokortikoidy, ale jsou navíc zatíženi častějšími nežádoucími účinky z důvodu užívání vyšších dávek glukokortikoidů. Klasifikace astmatu, od které se odvíjí stupňovité vedení komplexní léčebné strategie, není nadále jednotná a prochází neustálým vývojem. V následujícím přehledu bude kortikorezistentní astma definováno a zařazeno do aktuální doporučené české národní klasifikace astmatu. Budou uvedeny prokázané nebo předpokládané molekulární mechanizmy glukokortikoidní rezistence na buněčné úrovni u jedinců s kortikorezistentním astmatem.

Asthma is one of the most frequent chronic inflammatory disease amongst czech population. Generally asthma is caused by insufficient or inadequate regulation of systemic inflammatory activity manifested by chronic inflammatory changes in the airways of respiratory system. Complex treatment strategy, management approach based on treatment steps, is used in the treatment, with application of immunomodulatory drugs as a pharmacological base. Inhaled glucocorticoids are the drug of choice and concurrently the most often aplicated immunomodulatory drug amongst asthmatics. Glucocorticoid resistance amongst asthmatics – steroid (glucocorticosteroid) resistant asthma – is rather rarely. However, these steroid resistant asthmatics do not make a profit from the glucocorticoid therapy. Additionally, they suffer from more frequent side effects caused by taking higher doses of glucocorticoids. Steroid resistant asthma is exactly defined and classified. Classification of asthma, from which stepped management of complex treatment strategy is derived, is not yet unified and is still in the incessant process. In the following review steroid resistant asthma will be defined and classified according to actual czech national and international recommendations of asthma. Proven or expected molecular mechanisms of steroid resistant asthma on the cell level will be shown.

• Klíčová slova
• glukokortikoidní rezistence,
• MeSH
• biochemické jevy * MeSH
• bronchiální astma * farmakoterapie   klasifikace   metabolismus   MeSH
• glukokortikoidy * farmakologie   imunologie   MeSH
• léková rezistence * genetika   imunologie   MeSH
• lidé MeSH
• protein - isoformy MeSH
• receptory glukokortikoidů * genetika   metabolismus   MeSH
• transport proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

The protein homologous to the tumor suppressor p53, p73, has essential roles in development and tumorigenesis. This protein exists in a wide range of isoforms with different, even antagonistic, functions. However, there are virtually no detailed morphological studies analyzing the endogenous expression of p73 isoforms at the cellular level in cancer cells. In this study, we investigated the expression and subcellular distribution of two N-terminal isoforms, TAp73 and ΔNp73, in medulloblastoma cells using immunofluorescence microscopy. Both proteins were observed in all cell lines examined, but differences were noted in their intracellular localization between the reference Daoy cell line and four newly established medulloblastoma cell lines (MBL-03, MBL-06, MBL-07 and MBL-10). In the new cell lines, TAp73 and ΔNp73 were located predominantly in cell nuclei. However, there was heterogeneity in TAp73 distribution in the cells of all MBL cell lines, with the protein located in the nucleus and also in a limited non-random area in the cytoplasm. In a small percentage of cells, we detected cytoplasmic localization of TAp73 only, i.e., nuclear exclusion was observed. Our results provide a basis for future studies on the causes and function of distinct intracellular localization of p73 protein isoforms with respect to different protein-protein interactions in medulloblastoma cells.

• MeSH
• buněčné jádro metabolismus   MeSH
• dítě MeSH
• DNA vazebné proteiny chemie   metabolismus   MeSH
• intracelulární prostor metabolismus   MeSH
• jaderné proteiny chemie   metabolismus   MeSH
• lidé MeSH
• meduloblastom metabolismus   patologie   MeSH
• nádorové buněčné linie MeSH
• nádorové supresorové proteiny chemie   metabolismus   MeSH
• předškolní dítě MeSH
• protein - isoformy metabolismus   MeSH
• transport proteinů MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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

P53 controls the growth and survival of cells by acting in response to a multitude of cellular stresses. It is, however, not yet fully understood how different p53 activation pathways result in either cell cycle arrest or apoptosis. We and others have described an N-terminally truncated p53 protein (p53/47) originating from a second translation initiation site in the p53 messenger RNA (mRNA), which can interact with p53 and impose altered stability and transactivation properties to p53 complexes. Here we show that cap-dependent and cap-independent mechanisms of initiation govern the translation of the p53 mRNA. Changes in synthesis of full-length p53 or p53/47 are regulated through distinct cell stress-induced pathways acting through separate regions of the p53 mRNA. We also show that some cytotoxic drugs require the presence of full-length p53 to induce apoptosis, whereas for others p53/47 is sufficient. This indicates that by harbouring alternative translation initiation sites, the p53 mRNA gives rise to different levels of the p53 isoforms which help to orchestrate the cell biological outcome of p53 activation in response to different types of cell stress. This sheds new light into the way p53 can integrate and differentiate a large multiplicity of changes in the cellular environment.

• MeSH
• 5' nepřekládaná oblast MeSH
• exprese genu MeSH
• lidé MeSH
• messenger RNA * metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorový supresorový protein p53 * genetika   MeSH
• northern blotting MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein - isoformy * genetika   MeSH
• proteosyntéza * fyziologie   MeSH
• průtoková cytometrie MeSH
• regulace genové exprese * genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Expression of major cytochrome P450 forms (P450) was followed in preparation of purified hematopoietic CD34+ stem and progenitor cells. Levels of transcripts as well as mature proteins were traced by quantitative real-time polymerase chain reaction and by Northern and Western blotting. P450 1B1 and P450 2E1 proteins and respective mRNAs were found in all cases. On the other hand, no expression of P450 3A4, P450 3A7, and P450 2C9 was found. The results showed that expression of various P450 enzymes starts at different stages of cell differentiation. Both P450 forms found are known to be connected with cancer cells and with activation of procarcinogens (P450 1B1, polycyclic aromatic hydrocarbons; P450 2E1, nitrosamines, and solvents). Hence, cells at the early stage of differentiation already may be influenced by interaction with xenobiotics. This fact should also be taken into consideration when hematopoietic cell transplant therapy is applied.

• MeSH
• antigeny CD34 biosyntéza   MeSH
• cyklofilin A biosyntéza   genetika   MeSH
• exprese genu genetika   MeSH
• financování organizované MeSH
• hematopoetické kmenové buňky enzymologie   imunologie   metabolismus   MeSH
• imunoblotting MeSH
• izoenzymy biosyntéza   genetika   MeSH
• komplementární DNA izolace a purifikace   metabolismus   MeSH
• lidé MeSH
• messenger RNA biosyntéza   genetika   MeSH
• průtoková cytometrie MeSH
• RNA izolace a purifikace   metabolismus   MeSH
• systém (enzymů) cytochromů P-450 biosyntéza   genetika   MeSH
• Check Tag
• lidé MeSH

γ-Tubulin is the key protein for microtubule nucleation. Duplication of the γ-tubulin gene occurred several times during evolution, and in mammals γ-tubulin genes encode proteins which share ∼97% sequence identity. Previous analysis of Tubg1 and Tubg2 knock-out mice has suggested that γ-tubulins are not functionally equivalent. Tubg1 knock-out mice died at the blastocyst stage, whereas Tubg2 knock-out mice developed normally and were fertile. It was proposed that γ-tubulin 1 represents ubiquitous γ-tubulin, while γ-tubulin 2 may have some specific functions and cannot substitute for γ-tubulin 1 deficiency in blastocysts. The molecular basis of the suggested functional difference between γ-tubulins remains unknown. Here we show that exogenous γ-tubulin 2 is targeted to centrosomes and interacts with γ-tubulin complex proteins 2 and 4. Depletion of γ-tubulin 1 by RNAi in U2OS cells causes impaired microtubule nucleation and metaphase arrest. Wild-type phenotype in γ-tubulin 1-depleted cells is restored by expression of exogenous mouse or human γ-tubulin 2. Further, we show at both mRNA and protein levels using RT-qPCR and 2D-PAGE, respectively, that in contrast to Tubg1, the Tubg2 expression is dramatically reduced in mouse blastocysts. This indicates that γ-tubulin 2 cannot rescue γ-tubulin 1 deficiency in knock-out blastocysts, owing to its very low amount. The combined data suggest that γ-tubulin 2 is able to nucleate microtubules and substitute for γ-tubulin 1. We propose that mammalian γ-tubulins are functionally redundant with respect to the nucleation activity.

• MeSH
• časové faktory MeSH
• down regulace MeSH
• embryonální vývoj genetika   MeSH
• implantace embrya MeSH
• intracelulární prostor metabolismus   MeSH
• lidé MeSH
• mikrotubuly metabolismus   MeSH
• mitóza genetika   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• protein - isoformy nedostatek   genetika   metabolismus   MeSH
• transport proteinů MeSH
• tubulin nedostatek   genetika   metabolismus   MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

It has been suggested that thiazolidinediones (TZDs) ameliorate insulin resistance in muscle tissue by suppressing muscle lipid storage and the activity of novel protein kinase C (nPKC) isoforms. To test this hypothesis, we analyzed long-term metabolic effects of pioglitazone and the activation of nPKC-? and -? isoforms in an animal model of the metabolic syndrome, the spontaneously hypertensive rat (a congenic SHR strain with wild type Cd36 gene) fed a diet with 60 % sucrose from the age of 4 to 8 months. Compared to untreated controls, pioglitazone treatment was associated with significantly increased basal (809±36 vs 527±47 nmol glucose/g/2h, P<0.005) and insulinstimulated glycogenesis (1321±62 vs 749±60 nmol glucose/g/2h, P<0.0001) in isolated gastrocnemius muscles despite increased concentrations of muscle triglycerides (3.83±0.33 vs 2.25±0.12 µmol/g, P<0.005). Pioglitazone-treated rats exhibited significantly increased membrane/total (cytosolic plus membrane) ratio of both PKC-? and PKC-? isoforms compared to untreated controls. These results suggest that amelioration of insulin resistance after long-term pioglitazone treatment is associated with increased activation of PKC-? and -? isoforms in spite of increased lipid concentration in skeletal muscles.

• MeSH
• antigeny CD36 genetika   metabolismus   MeSH
• časové faktory MeSH
• financování organizované MeSH
• glykogen metabolismus   MeSH
• hypoglykemika farmakologie   MeSH
• inzulin metabolismus   MeSH
• inzulinová rezistence MeSH
• izoenzymy metabolismus   MeSH
• konzumní sacharóza aplikace a dávkování   metabolismus   MeSH
• kosterní svaly enzymologie   patofyziologie   účinky léků   MeSH
• krevní glukóza metabolismus   MeSH
• krysa rodu rattus MeSH
• metabolický syndrom MeSH
• modely nemocí na zvířatech MeSH
• potkani inbrední SHR MeSH
• proteinkinasa C metabolismus   MeSH
• thiazolidindiony farmakologie   MeSH
• transport proteinů MeSH
• triglyceridy metabolismus   MeSH
• zvířata kongenní MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH

Recognition of glycosylation patterns is one of the basic features of innate immunity. Ability of C-type lectin-like receptors such as NKR-P1 to bind saccharide moieties has become recently a controversial issue. In the present study, binding assay with soluble fluorescently labeled recombinant rat NKR-P1A and mouse NKR-P1C proteins revealed apparently no affinity to the various neoglycoproteins. Lack of functional linkage between NKR-P1 and previously described saccharide binder was supported by the fact, that synthetic N-acetyl-D-glucosamine octabranched dendrimer on polyamidoamine scaffold (GN8P) did not change gene expression of NKR-P1 isoforms in C57BL/6 and BALB/c mice divergent in the NK gene complex (both in vitro and in vivo). Surprisingly, N-acetyl-D-glucosamine-coated tetrabranched polyamido-amine dendrimer specifically binds to NKT cells and macrophages but not to NK cells (consistently with changes in cytokine patterns). Despite the fact that GN8P has been tested as an immunomodulator in anti-cancer treatment animal models for many years, surprisingly no changes in cytokine profiles in serum relevant to anti-cancer responses using B16F10 and CT26 harboring mouse strains C57BL/6 and BALB/c are observed. Our results indicate possible indirect involvement of NK cells in GN8P mediated immune responses.

• MeSH
• acetylglukosamin imunologie   metabolismus   MeSH
• buňky NK imunologie   metabolismus   MeSH
• dendrimery metabolismus   MeSH
• experimentální nádory farmakoterapie   genetika   imunologie   MeSH
• exprese genu účinky léků   imunologie   MeSH
• glykokonjugáty imunologie   metabolismus   farmakologie   MeSH
• interferon gama krev   genetika   imunologie   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• lektinové receptory NK-buněk - podrodina B genetika   imunologie   metabolismus   MeSH
• lektiny typu C genetika   imunologie   metabolismus   MeSH
• makrofágy imunologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• NKT buňky imunologie   metabolismus   MeSH
• oligosacharidy imunologie   metabolismus   MeSH
• polyaminy imunologie   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein - isoformy genetika   imunologie   metabolismus   MeSH
• průtoková cytometrie MeSH
• slezina cytologie   imunologie   metabolismus   MeSH
• TNF-alfa krev   genetika   imunologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The platinum(II)-based complex cisplatin is one of the most frequently used antitumour agents; however, a high incidence of harmful side effects and the frequent emergence of acquired resistance are the major clinical problems. The novel platinum(IV)-based complex LA-12 exhibits a high efficacy against cancer cell lines, including cisplatin-insensitive cells, but the mechanisms by which LA-12 perturbs cell growth are unclear. We tested the effects of LA-12 on the p53 response and demonstrate that LA-12 induces unique changes in the profile of gene expression compared with cisplatin and doxorubicin. Furthermore, the ability of LA-12 to disrupt cellular proliferation is greatly enhanced by the expression of p53 and p53/47 indicating both p53-dependent and p53-independent effects of LA-12. Exposure of the human cancer cell lines H1299, A2780, BT549 and BT474 to LA-12 alters the expression of p53 and p53/47 in both a time-dependent and dose-dependent manner. Treatment of cells with a low concentration of the drug results in accumulation of p53 and p53/47 concomitant with their posttranslational modification, whereas a high dose results in the disappearance of both the forms of p53. The distinct p53 activation profile of LA-12 compared with cisplatin and doxorubicin provides a molecular explanation for the ability of this drug to treat cisplatin-resistant cells and indicates its potential usefulness as an alternative antitumour agent in first-line therapy or as a second-line therapy in patients with acquired cisplatin resistance.

• MeSH
• amantadin analogy a deriváty   farmakologie   MeSH
• buněčný cyklus účinky léků   MeSH
• chemorezistence MeSH
• cisplatina farmakologie   MeSH
• doxorubicin MeSH
• financování organizované MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• organoplatinové sloučeniny farmakologie   MeSH
• posttranslační úpravy proteinů MeSH
• proliferace buněk účinky léků   MeSH
• protein - isoformy genetika   metabolismus   MeSH
• protinádorové látky farmakologie   MeSH
• průtoková cytometrie MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• signální transdukce MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• srovnávací studie MeSH

Stanovit solubilní a na buňky vázané angiogenní faktory odrážející růstovou aktivitu karcinomu ledviny a stanovit prognostický význam. Pomocí vybraných parametrů (VEGF, bFGF, TGF a CD105) stanovit riziko progrese karcinomu ledviny v čase stanovení diagnózy. Pomocí průtokové cytometrie stanovit expresi CD105 (endoglin) na nádorových buňkách a buňkách cévních epitelií. Zjistit, zda opakovaná stanovení hladin angiogenních faktorů mohou odhalit progresi karcinomu ledviny.; To verify the soluble and membrane associated angiogenic factors reflected the growth activity of renal cell carcinoma and set up there prognostic value. To pick out some parameters (VEGF, bFGF, TGF, CD105) to determine the risk of progression of RCC intime of diagnosis. To use the flow cytometry for detection of CD105 expression on membranes of tumor and endothelial vessel cells. To verify the diagnostic importance of repeated detection of angiogenic factors during the relaps of disease.

• MeSH
• biologické markery analýza   MeSH
• erbB receptory MeSH
• fibroblastový růstový faktor 2 MeSH
• intracelulární signální peptidy a proteiny diagnostické užití   MeSH
• karcinom z renálních buněk MeSH
• látky indukující angiogenezi MeSH
• protein - isoformy diagnostické užití   MeSH
• průtoková cytometrie MeSH
• vaskulární endoteliální růstové faktory MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• nefrologie
• onkologie
• biologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR