MicroRNAs (miRNAs) are important regulators of heart function and then an intriguing therapeutic target for plenty of diseases. The problem raised is that many data in this area are contradictory, thus limiting the use of miRNA-based therapy. The goal of this review is to describe the hub-mechanisms regulating the biogenesis and function of miRNAs, which could help in clarifying some contradictions in the miRNA world. With this scope, we analyse an array of factors, including several known agents of stress response, mediators of epigenetic changes, regulators of alternative splicing, RNA editing, protein synthesis and folding and proteolytic systems. All these factors are important in cardiovascular function and most of them regulate miRNA biogenesis, but their influence on miRNAs was shown for non-cardiac cells or some specific cardiac pathologies. Finally, we consider that studying the stress response factors, which are upstream regulators of miRNA biogenesis, in the diseased heart could help in (1) explaining some contradictions concerning miRNAs in heart pathology, (2) making the role of miRNAs in pathogenesis of cardiovascular disease more clear, and therefore, (3) getting powerful targets for its molecular therapy.

• Klíčová slova
• cardiac pathophysiology, hub-mechanisms, miRNAs biogenesis, stress response,
• MeSH
• biologické modely MeSH
• epigeneze genetická MeSH
• fyziologický stres genetika   MeSH
• kardiovaskulární nemoci genetika   MeSH
• lidé MeSH
• mikro RNA biosyntéza   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• mikro RNA MeSH

Adipose tissue is an abundant source of autologous adult stem cells that may bring new therapeutic perspectives on the treatment of diabetes and its complications. It is unclear whether adipose tissue-derived stromal cells (ASCs) of diabetic patients, constantly influenced by hyperglycaemia, have the same properties as non-diabetic controls. As an alternative source of ASCs, adipose tissue from distal limbs of diabetic patients with critical ischemia was isolated. ASCs were characterized in terms of cell surface markers, multilineage differentiation and the expression of vascular endothelial growth factor (VEGFA), chemokine-related genes and compared with non-diabetic controls. Flow cytometry analysis confirmed mesenchymal phenotypes in both diabetic and non-diabetic ASCs. Nevertheless, 40% of diabetic and 20% of non-diabetic ASC samples displayed high expressions of fibroblast marker, which inversely correlated with the expression of CD105. In diabetic patients, significantly decreased expression of VEGFA and chemokine receptor CXCR4 was found in fibroblast-positive ASCs, compared with their fibroblast-negative counterparts. Reduced osteogenic differentiation and the downregulation of chemokine CXCL12 were found in fibroblast-negative diabetic ASCs. Both diabetic and non-diabetic ASCs were differentiated into adipocytes and chondrocytes and did not reveal islet-like cell differentiation. According to this study, adipose tissue from distal limbs of diabetic patients is not satisfactory as an autologous ASC source. Hyperglycaemic milieu as well as other metabolic disorders linked to diabetes may have an influence on endogenous stem cell properties. The present study investigated the feasibility of autologous stem cell therapy in diabetic patients. ASCs isolated from the ischemic limb of diabetic patients were found to be less potent when compared phenotypically and functionally to control non-diabetic counterparts with no signs of limb ischemia. High expression of fibroblast markers associated with reduced expression of VEGFA as well as reduced osteogenic differentiation may have an impact on the effectiveness of autologous cell therapies in diabetic patients.

• Klíčová slova
• adipose tissue, cell therapy, diabetes, differentiation, flow cytometry, mesenchymal stem cells,
• MeSH
• biologické markery metabolismus   MeSH
• buněčná diferenciace MeSH
• buňky stromatu cytologie   metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• chondrocyty cytologie   metabolismus   MeSH
• cytokiny metabolismus   MeSH
• diabetes mellitus patologie   MeSH
• diabetická noha patologie   MeSH
• dospělí MeSH
• endoglin MeSH
• fibroblasty cytologie   metabolismus   MeSH
• ischemie patologie   MeSH
• končetiny krevní zásobení   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• osteoblasty cytologie   metabolismus   MeSH
• podkožní tuk cytologie   MeSH
• receptory buněčného povrchu metabolismus   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• tukové buňky cytologie   metabolismus   MeSH
• vaskulární endoteliální růstový faktor A metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• CD antigeny MeSH
• cytokiny MeSH
• endoglin MeSH
• ENG protein, human MeSH Prohlížeč
• receptory buněčného povrchu MeSH
• vaskulární endoteliální růstový faktor A MeSH

Alzheimer's disease (AD) is accompanied by oxidative stress in the brain. Because the brain tissue is rich in polyunsaturated fatty acids, it is prone to the free radical attack resulting in lipid peroxidation. Intermediates of lipid peroxidation may diffuse from the primary site, cross the blood-brain barrier and modify erythrocyte membranes in the bloodstream. We exposed isolated erythrocyte membranes from patients with AD and the control group to in vitro free radical damage and monitored the accumulation of the end products of lipid peroxidation, lipofuscin-like pigments (LFPs), by fluorescence spectroscopy. LFPs were analyzed by means of tridimensional and synchronous fluorescence spectroscopy. The levels of LFP formed during in vitro peroxidation were significantly higher in erythrocyte membranes from patients with AD compared with the control group. Furthermore, the chemical composition of LFP in AD was different from the control group. The analysis of the specific modifications of erythrocyte membranes in AD is of great medical importance regarding the need of a diagnostic blood biomarker.

• MeSH
• Alzheimerova nemoc metabolismus   MeSH
• erytrocytární membrána metabolismus   MeSH
• lidé MeSH
• lipofuscin metabolismus   MeSH
• oxidační stres MeSH
• peroxidace lipidů * MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• lipofuscin MeSH

We have examined the effect of suberoylanilide hydroxamic acid (SAHA, Vorinostat, Zolinza) on the viability of normal peripheral blood lymphocytes (PBLs) in vitro and on the expression of 20 apoptosis-related genes. RT-PCR, western blots and flow cytometry were performed to reveal the proteins of apoptosis machinery that were affected to cause cell death. Our data suggest that PBL markedly resisted for approximately 24 h the destructive activity of the agent, but eventually 60% of cells treated with 4 micromol/L SAHA died within 72 h through mitochondrial way of apoptosis. While the expression of the majority of genes remained indifferent against 4 micromol/L SAHA, the cellular levels of BimEL, Bmf-2, Bcl-w and survivin mRNA varied, confirming the pro-apoptotic response of SAHA treated PBL. In addition, the expression of multifunctional proteins c-Myc and p21(WAF1) changed profoundly with the time of SAHA treatment. The Bax activator BimEL increased rapidly, driving cells towards apoptosis likely controlled by c-Myc and p21(WAF1) activities. We suggest that variations in c-Myc and p21(WAF1) expression decelerate the apoptosis in the early period and increase the resistance of resting PBL against SAHA.

• MeSH
• aktivace enzymů účinky léků   MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• buněčná smrt účinky léků   MeSH
• cytochromy c metabolismus   MeSH
• cytoprotekce * účinky léků   MeSH
• fosfatidylseriny metabolismus   MeSH
• inhibitor p21 cyklin-dependentní kinasy metabolismus   MeSH
• kaspasy metabolismus   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• lidé MeSH
• lymfocyty cytologie   účinky léků   enzymologie   metabolismus   MeSH
• membránové proteiny metabolismus   MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• mitochondrie účinky léků   metabolismus   MeSH
• poly(ADP-ribosa)polymerasy metabolismus   MeSH
• protein BCL2L11 MeSH
• proteiny regulující apoptózu metabolismus   MeSH
• protoonkogenní proteiny c-myc metabolismus   MeSH
• protoonkogenní proteiny metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• transport proteinů účinky léků   MeSH
• viabilita buněk účinky léků   MeSH
• vorinostat MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• BCL2L11 protein, human MeSH Prohlížeč
• cytochromy c MeSH
• fosfatidylseriny MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• kaspasy MeSH
• kyseliny hydroxamové MeSH
• membránové proteiny MeSH
• poly(ADP-ribosa)polymerasy MeSH
• protein BCL2L11 MeSH
• proteiny regulující apoptózu MeSH
• protoonkogenní proteiny c-myc MeSH
• protoonkogenní proteiny MeSH
• reaktivní formy kyslíku MeSH
• vorinostat MeSH

We studied changes in antioxidant protection during ageing and senescence in chloroplasts of tobacco (Nicotiana tabacum L., cv. Wisconsin) with introduced SAG(12) promoter fused with ipt gene for cytokinin synthesis (transgenic plants with increased levels of cytokinins, SAG) or without it (control). Old leaves of SAG plants as well as their chloroplasts maintained higher physiological parameters compared to controls; accordingly, we concluded that their ageing was diverted due to increased cytokinin content. The chloroplast antioxidant protection did not decrease as well. Although antioxidant protection usually decreased in whole leaves of senescing control plants, ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR) activity, which maintained the high redox state of ascorbate, increased in chloroplasts of old control leaves.

• MeSH
• antioxidancia fyziologie   MeSH
• časové faktory MeSH
• chloroplasty metabolismus   MeSH
• cytokininy biosyntéza   MeSH
• geneticky modifikované rostliny cytologie   genetika   fyziologie   MeSH
• listy rostlin metabolismus   MeSH
• stárnutí buněk fyziologie   MeSH
• tabák cytologie   genetika   metabolismus   fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• antioxidancia MeSH
• cytokininy MeSH

The molecular mechanisms involved in GPCR-initiated signaling cascades where the two receptors share the same signaling cascade, such as thyrotropin-releasing hormone (TRH) and angiotensin II (ANG II), are still far from being understood. Here, we analyzed hormone-induced Ca(2+) responses and the process of desensitization in HEK-293 cells, which express endogenous ANG II receptors. These cells were transfected to express exogenously high levels of TRH receptors (clone E2) or both TRH receptors and G(11)alpha protein (clone E2M11). We observed that the characteristics of the Ca(2+) response, as well as the process of desensitization, were both strongly dependent on receptor number and G(11)alpha protein level. Whereas treatment of E2 cells with TRH or ANG II led to significant desensitization of the Ca(2+) response to subsequent addition of either hormone, the response was not desensitized in E2M11 cells expressing high levels of G(11)alpha. In addition, stimulation of both cell lines with THR elicited a clear heterologous desensitization to subsequent stimulation with ANG II. On the other hand, ANG II did not affect a subsequent response to TRH. ANG II-mediated signal transduction was strongly dependent on plasma membrane integrity modified by cholesterol depletion, but signaling through TRH receptors was altered only slightly under these conditions. It may be concluded that the level of expression of G-protein-coupled receptors and their cognate G-proteins strongly influences not only the magnitude of the Ca(2+) response but also the process of desensitization and resistance to subsequent hormone addition.

• MeSH
• angiotensin II farmakologie   MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• časové faktory MeSH
• cholesterol analýza   metabolismus   MeSH
• hormon uvolňující thyreotropin farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• proteiny vázající GTP - alfa-podjednotky Gq-G11 biosyntéza   účinky léků   genetika   MeSH
• receptory thyroliberinu biosyntéza   účinky léků   MeSH
• teplota MeSH
• transfekce MeSH
• vápník metabolismus   farmakologie   MeSH
• vápníková signalizace účinky léků   fyziologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• angiotensin II MeSH
• cholesterol MeSH
• hormon uvolňující thyreotropin MeSH
• proteiny vázající GTP - alfa-podjednotky Gq-G11 MeSH
• receptory thyroliberinu MeSH
• vápník MeSH

Several studies report that hypoxic exposure induces free radical oxidative damage in various tissues. The mechanism of this damage includes membrane lipid peroxidation which can be easily detected by measuring fluorescent end-products of the process, i.e. lipofuscin-like pigments. Four day exposure of rats to hypoxia (10% O(2)) increased the level of lipofuscin-like pigments in erythrocytes up to 9 fold. This increase was completely prevented when the animals were exposed to hypercapnia (4.3% CO(2)) in addition to hypoxia. We studied the possible mechanism of the hypercapnic protection on isolated erythrocyte membranes in vitro. Lipid peroxidation was initiated by incubation of the membranes with iron ions and ascorbate. Production of malonaldehyde, the precursor of lipofuscin-like pigments, was strongly inhibited in bicarbonate buffer. Similarly the production of lipofuscin-like products was damped. These experiments suggest that the protective effect of hypercapnia might consist in direct interaction of CO(2) with free radical processes.

• MeSH
• časové faktory MeSH
• cytoprotekce * MeSH
• erytrocytární membrána metabolismus   MeSH
• erytrocyty cytologie   metabolismus   MeSH
• fluorescenční spektrometrie MeSH
• hyperkapnie metabolismus   MeSH
• hypoxie metabolismus   MeSH
• krysa rodu Rattus MeSH
• lipofuscin biosyntéza   MeSH
• luminiscence MeSH
• luminol metabolismus   MeSH
• malondialdehyd metabolismus   MeSH
• peroxidace lipidů MeSH
• potkani Wistar MeSH
• volné radikály metabolismus   MeSH
• železo metabolismus   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
• lipofuscin MeSH
• luminol MeSH
• malondialdehyd MeSH
• volné radikály MeSH
• železo MeSH

We have examined the effect of sodium butyrate (SB) on the viability of normal peripheral blood lymphocytes (PBLs) in vitro and the effect of this agent on the expression of 20 apoptosis-related genes. Data suggest that PBL treated with 2 mmol L(-1) SB resisted for at least 8 h the destructive activity of the agent, but eventually 30% of cells died within 72 h. As documented by flow cytometry and cytochrome c release study, cells underwent mitochondrial-derived apoptosis. While the expression of the majority of genes examined by RT-PCR and Western blots remained indifferent to 2 mmol L(-1) SB, the cellular levels of BimEL, c-myc, p53, and p21(WAF1) varied profoundly with the time of SB treatment. The Bax activator BimEL increased rapidly, driving cells toward apoptosis likely controlled by c-myc and p21(WAF1) activities. The c-myc, exercising the role of mediator of the function of BimEL and inhibitor of p21(WAF1) expression, decreased significantly for several hours after adding SB but within 48 h it returned to close to its original value. An apoptosis inhibitor and executive caspase substrate p21(WAF1) increased early at the beginning of treatment but subsequently, within a time frame of 72 h, profoundly dropped in terms of both a caspase-dependent and caspase-independent way. We suggest that variations in c-myc and p21(WAF1) expression delay apoptosis making PBL resistant to SB for several hours, and together with fast catabolism of SB in vivo protect PBL against the destructive activity of this anti-cancerous metabolite of colonic bacteria.

• MeSH
• aktivace enzymů MeSH
• apoptóza účinky léků   MeSH
• cytochromy c metabolismus   MeSH
• inhibitor p21 cyklin-dependentní kinasy genetika   metabolismus   MeSH
• intracelulární membrány účinky léků   MeSH
• kaspasy metabolismus   MeSH
• kolagen typ XI metabolismus   MeSH
• kultivované buňky MeSH
• kyselina máselná farmakologie   MeSH
• lidé MeSH
• lymfocyty účinky léků   metabolismus   MeSH
• membránové proteiny genetika   metabolismus   MeSH
• mitochondrie účinky léků   MeSH
• protein BCL2L11 MeSH
• proteiny regulující apoptózu genetika   metabolismus   MeSH
• protoonkogenní proteiny c-myc genetika   metabolismus   MeSH
• protoonkogenní proteiny genetika   metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• BCL2L11 protein, human MeSH Prohlížeč
• Bcl2l11 protein, mouse MeSH Prohlížeč
• CDKN1A protein, human MeSH Prohlížeč
• COL11A2 protein, human MeSH Prohlížeč
• cytochromy c MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• kaspasy MeSH
• kolagen typ XI MeSH
• kyselina máselná MeSH
• membránové proteiny MeSH
• MYC protein, human MeSH Prohlížeč
• Perp protein, mouse MeSH Prohlížeč
• protein BCL2L11 MeSH
• proteiny regulující apoptózu MeSH
• protoonkogenní proteiny c-myc MeSH
• protoonkogenní proteiny MeSH
• reaktivní formy kyslíku MeSH

CYP2D6 is a member of cytochrome P450 enzymes that metabolise over 25% of commonly used drugs. Genetic polymorphisms can cause insufficient drug efficacy at usually administered doses or can be the cause of adverse drug reaction. CYP2D6 genotyping can be used to predict CYP2D6 phenotype and thereby explain some abnormalities in drug response and thus optimize pharmacotherapy. The aim of this study was to investigate the frequency of functionally important variant alleles of the CYP2D6 gene throughout the Czech population to predict the prevalence of ultra-rapid and poor metabolizer phenotypes. The DNA of 223 unrelated, healthy volunteers was analysed to detect the presence of CYP2D6*6, *5, *4, *3 and gene duplication. The variant allele frequencies in our population were 0.22%, 3.14%, 22.87%, 1.12% and 3.14% for CYP2D6*6, CYP2D6*5, CYP2D6*4, CYP2D6*3 and CYP2D6*MxN, respectively. Fifteen subjects carried two variant alleles leading to predicted poor type of metabolism, 84 subjects were heterozygous extensive metabolizers (het-EM). The full-text contains detailed comparison with European white populations. The distribution of variant alleles complies with the Hardy-Weinberg equilibrium. The frequencies of functional variant alleles of CYP2D6 in Czech population are in concordance with other Caucasian populations.

• MeSH
• cytochrom P-450 CYP2D6 genetika   fyziologie   MeSH
• dospělí MeSH
• farmakogenetika MeSH
• frekvence genu * MeSH
• genotyp MeSH
• jednonukleotidový polymorfismus * MeSH
• léčivé přípravky metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• populační genetika MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• cytochrom P-450 CYP2D6 MeSH
• léčivé přípravky MeSH

The aim of the present work was to evaluate non-enzymic antioxidants during natural and artificially modulated senescence. Senescence of bean (Phaseolus vulgaris L. cv. Jantar) cotyledons was modulated by UV C irradiation or by the decapitation of plants apices. The content of beta-carotene and zeaxanthin decreased in control and decapitated plants but in UV C irradiated plants these contents increased. The degree of de-epoxidation increased in all cultivations with age. The content of total glutathione (sum of reduced and oxidized) sharply decreased in bean cotyledons grown in all conditions. Interestingly, the content of total ascorbate increased at the end of cotyledon life span of control and decapitated plants but decreased in UV plants. Decrease of reduced/oxidized ratio of ascorbate and glutathione during cotyledon ageing confirmed increasing oxidative stress during senescence in all cultivations.

• MeSH
• antioxidancia metabolismus   MeSH
• beta-katenin metabolismus   MeSH
• epoxidové sloučeniny metabolismus   MeSH
• fazol růst a vývoj   metabolismus   MeSH
• molekulová hmotnost MeSH
• oxidace-redukce MeSH
• xanthofyly metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antioxidancia MeSH
• beta-katenin MeSH
• epoxidové sloučeniny MeSH
• xanthofyly MeSH