Nitric oxide (NO) stimulated the activity of plasma membrane H+-ATPase, 5'-nucleotidase, peroxidase, ascorbate peroxidase and glutathione reductase in ultraviolet B (UV-B) irradiated Chlorella pyrenoidosa. It also boosted the activity of nitrogen-metabolism enzymes such as nitrate reductase, nitrite reductase, glutamine synthetase, which were inhibited by UV-B irradiation. The chlorophyll fluorescence ratio (Fv/Fm) of the UV-B irradiated algae and decreased continuously after the cells were transferred to UV-B irradiation. A continuing decrease of the Fv/Fm was observed even after the cells were transferred to photosynthetically active radiation (PAR). After adaptation for 8 h under PAR (after treatment with nitric oxide), Fv/Fm recovered to 55 % of normal levels--without NO the value approached zero. Exogenous NO stopped the decay of chlorophyll and thylakoid membrane in cells exposed to UV-B irradiation. NO plays probably a key role in damage induced by UV-B irradiation in green algae.

• MeSH
• bílkoviny řas metabolismus   MeSH
• buněčná membrána enzymologie   MeSH
• Chlorella fyziologie   účinky záření   MeSH
• enzymy metabolismus   MeSH
• financování organizované MeSH
• oxid dusnatý fyziologie   MeSH
• systémy druhého messengeru MeSH
• ultrafialové záření MeSH

• MeSH
• energetický metabolismus fyziologie   MeSH
• experimenty na zvířatech MeSH
• finanční podpora výzkumu jako téma MeSH
• guanosinmonofosfát cyklický fyziologie   MeSH
• kosterní svaly metabolismus   MeSH
• oxid dusnatý fyziologie   MeSH
• potkani Wistar MeSH
• regionální krevní průtok MeSH
• spotřeba kyslíku fyziologie   MeSH
• systémy druhého messengeru fyziologie   MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

We investigated the influence of membrane cholesterol content on preferential and non-preferential signaling through the M(2) muscarinic acetylcholine receptor expressed in CHO cells. Cholesterol depletion by 39% significantly decreased the affinity of M(2) receptors for [(3)H]-N-methylscopolamine ([(3)H]-NMS) binding and increased B(max) in intact cells and membranes. Membranes displayed two-affinity agonist binding sites for carbachol and cholesterol depletion doubled the fraction of high-affinity binding sites. In intact cells it also reduced the rate of agonist-induced receptor internalization and changed the profile of agonist binding from a single site to two affinity states. Cholesterol enrichment by 137% had no effects on carbachol E(max) of cAMP synthesis inhibition and on cAMP synthesis stimulation and inositolphosphates (IP) accumulation at higher agonist concentrations (non-preferred pathways). On the other hand, cholesterol depletion significantly increased E(max) of cAMP synthesis inhibition or stimulation without change in potency, and decreased E(max) of IP accumulation. Noteworthy, modifications of membrane cholesterol had no effect on membrane permeability, oxidative activity, protein content, or relative expression of G(s), G(i/o), and G(q/11) alpha subunits. These results demonstrate distinct changes of M(2) receptor signaling through both preferential and non-preferential G-proteins consequent to membrane cholesterol depletion that occur at the level of receptor/G-protein/effector protein interactions in the cell membrane. The significant decrease of IP accumulation by cholesterol depletion was also observed in cells expressing M(3) receptors and by both cholesterol depletion and enrichment in cells expressing M(1) receptors indicating relevance of reduced G(q/11) signaling for the pathogenesis of Alzheimer's disease.

• MeSH
• acetylcholin analogy a deriváty   MeSH
• antagonisté muskarinových receptorů farmakologie   MeSH
• beta-cyklodextriny farmakologie   MeSH
• buněčná membrána metabolismus   účinky léků   MeSH
• CHO buňky MeSH
• cholesterol metabolismus   MeSH
• Cricetulus MeSH
• karbachol analogy a deriváty   farmakologie   metabolismus   MeSH
• křečci praví MeSH
• lidé MeSH
• N-methylskopolamin farmakologie   metabolismus   MeSH
• proteiny vázající GTP metabolismus   MeSH
• receptor muskarinový M2 metabolismus   MeSH
• systémy druhého messengeru účinky léků   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural 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

• MeSH
• finanční podpora výzkumu jako téma MeSH
• krysa rodu rattus MeSH
• svaly krevní zásobení   MeSH
• systémy druhého messengeru MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

UNLABELLED: Obesity and metabolic syndrome is increasing health problem worldwide. Among other ways, nutritional intervention using phytochemicals is important method for treatment and prevention of this disease. Recent studies have shown that certain phytochemicals could alter the expression of specific genes and microRNAs (miRNAs) that play a fundamental role in the pathogenesis of obesity. For study of the obesity and its treatment, monosodium glutamate (MSG)-injected mice with developed central obesity, insulin resistance and liver lipid accumulation are frequently used animal models. To understand the mechanism of phytochemicals action in obese animals, the study of selected genes expression together with miRNA quantification is extremely important. For this purpose, real-time quantitative PCR is a sensitive and reproducible method, but it depends on proper normalization entirely. The aim of present study was to identify the appropriate reference genes for mRNA and miRNA quantification in MSG mice treated with green tea catechins, potential anti-obesity phytochemicals. Two sets of reference genes were tested: first set contained seven commonly used genes for normalization of messenger RNA, the second set of candidate reference genes included ten small RNAs for normalization of miRNA. The expression stability of these reference genes were tested upon treatment of mice with catechins using geNorm, NormFinder and BestKeeper algorithms. Selected normalizers for mRNA quantification were tested and validated on expression of NAD(P)H: quinone oxidoreductase, biotransformation enzyme known to be modified by catechins. The effect of selected normalizers for miRNA quantification was tested on two obesity- and diabetes- related miRNAs, miR-221 and miR-29b, respectively. Finally, the combinations of B2M/18S/HPRT1 and miR-16/sno234 were validated as optimal reference genes for mRNA and miRNA quantification in liver and 18S/RPlP0/HPRT1 and sno234/miR-186 in small intestine of MSG mice. These reference genes will be used for mRNA and miRNA normalization in further study of green tea catechins action in obese mice.

• MeSH
• algoritmy MeSH
• geny * MeSH
• kvantitativní polymerázová řetězová reakce metody   normy   MeSH
• messenger RNA genetika   metabolismus   MeSH
• mikro RNA genetika   metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• obezita genetika   MeSH
• referenční standardy MeSH
• regulace genové exprese MeSH
• reprodukovatelnost výsledků MeSH
• software MeSH
• stanovení celkové genové exprese MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• adenylátcyklasy fyziologie   MeSH
• autoradiografie MeSH
• fosfatidylinositoly fyziologie   MeSH
• neurochemie analýza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

• MeSH
• fosfatidylinositoly genetika   metabolismus   MeSH
• vápníkové kanály fyziologie   MeSH
• Publikační typ
• přehledy MeSH

• MeSH
• AMP cyklický fyziologie   MeSH
• buněčná membrána MeSH
• hormony fyziologie   sekrece   MeSH
• kyseliny fosfatidové analogy a deriváty   fyziologie   MeSH
• receptory buněčného povrchu fyziologie   MeSH

We report a case of a 52-year-old female with synovial sarcoma of the uterine corpus. Grossly, the partly polypoid tumor involved the endometrium with invasion into the inner half of the myometrium. Histologically, the tumor showed biphasic structure with the predominance of poorly differentiated small to medium sized round to oval cells. These cells showed high nuclear to cytoplasmic ratio and were arranged in diffuse sheets. Other component consisted of larger epitheloid cells with ample eosinophilic cytoplasm arranged in irregular nests. These cells were only present in a small amount. Immunohistochemically, the tumor cells in both components showed the expression of EMA, S-100 protein, CD99, and NSE. RT-PCR analysis showed the presence of SYT-SSX1 fusion transcript. At present, the patient shows no signs of tumor relapse 56 months after the diagnosis. To the best of our knowledge, this is the first report of synovial sarcoma arising in uterus.

• MeSH
• fúzní onkogenní proteiny genetika   MeSH
• imunoenzymatické techniky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• nádory děložního čípku genetika   patologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• sekundární malignity genetika   patologie   MeSH
• synoviom genetika   patologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH