Although the modulation of host physiology has been interpreted as an essential process supporting baculovirus propagation, the requirement of energy supply for host antivirus reactions could not be ruled out. Our present study showed that metabolic induction upon AcMNPV (budded virus) infection of Bombyx mori stimulated virus clearance and production of the antivirus protein, gloverin. In addition, we demonstrated that adenosine receptor signaling (AdoR) played an important role in regulating such metabolic reprogramming upon baculovirus infection. By using a second lepidopteran model, Spodoptera frugiperda Sf-21 cells, we demonstrated that the glycolytic induction regulated by adenosine signaling was a conservative mechanism modulating the permissiveness of baculovirus infection. Another interesting finding in our present study is that both BmNPV and AcMNPV infection cause metabolic activation, but it appears that BmNPV infection moderates the level of ATP production, which is in contrast to a dramatic increase upon AcMNPV infection. We identified potential AdoR miRNAs induced by BmNPV infection and concluded that BmNPV may attempt to minimize metabolic activation by suppressing adenosine signaling and further decreasing the host's anti-baculovirus response. Our present study shows that activation of energy synthesis by adenosine signaling upon baculovirus infection is a host physiological response that is essential for supporting the innate immune response against infection.

• MeSH
• adenosin metabolismus   MeSH
• adenosintrifosfát biosyntéza   MeSH
• bourec metabolismus   virologie   MeSH
• deoxyglukosa farmakologie   MeSH
• energetický metabolismus MeSH
• glykolýza účinky léků   genetika   MeSH
• hmyzí proteiny metabolismus   MeSH
• infekce DNA virem metabolismus   virologie   MeSH
• interakce hostitele a patogenu imunologie   MeSH
• mezibuněčné signální peptidy a proteiny metabolismus   MeSH
• nukleopolyhedroviry fyziologie   MeSH
• purinergní receptory P1 genetika   metabolismus   MeSH
• replikace viru účinky léků   MeSH
• Sf9 buňky MeSH
• Spodoptera MeSH
• transfekce MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Adenosine (Ado) is a ubiquitous metabolite that plays a prominent role as a paracrine homeostatic signal of metabolic imbalance within tissues. It quickly responds to various stress stimuli by adjusting energy metabolism and influencing cell growth and survival. Ado is also released by dead or dying cells and is present at significant concentrations in solid tumors. Ado signaling is mediated by Ado receptors (AdoR) and proteins modulating its concentration, including nucleoside transporters and Ado deaminases. We examined the impact of genetic manipulations of three Drosophila genes involved in Ado signaling on the incidence of somatic mosaic clones formed by the loss of heterozygosity (LOH) of tumor suppressor and marker genes. We show here that genetic manipulations with the AdoR, equilibrative nucleoside transporter 2 (Ent2), and Ado deaminase growth factor-A (Adgf-A) cause dramatic changes in the frequency of hyperplastic outgrowth clones formed by LOH of the warts (wts) tumor suppressor, while they have almost no effect on control yellow (y) clones. In addition, the effect of AdoR is dose-sensitive and its overexpression leads to the increase in wts hyperplastic epithelial outgrowth rates. Consistently, the frequency of mosaic hyperplastic outgrowth clones generated by the LOH of another tumor suppressor, discs overgrown (dco), belonging to the wts signaling pathway is also dependent on AdoR. Our results provide interesting insight into the maintenance of tissue homeostasis at a cellular level.

• MeSH
• Drosophila melanogaster MeSH
• membránové transportní proteiny genetika   MeSH
• mutace * MeSH
• proteiny Drosophily genetika   MeSH
• purinergní receptory P1 genetika   MeSH
• signální transdukce genetika   MeSH
• ztráta heterozygozity MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Gut ischemia and reperfusion (IR), e.g. in small bowel transplantation or during resuscitation, may result in severe impairment of the intestinal microcirculation. Potential sequelae are mucosal damage, loss of intestinal barrier function, bacterial translocation, systemic inflammation, multiple organ failure and death. We hypothesized a protective role for extracellular adenosine signalling in intestinal IR injury. Using intravital microscopy we investigated the effects of the adenosine receptor (AR) agonist NECA (5'-N-ethyl carboxamide adenosine) on leukocyte-endothelial interactions and capillary perfusion in the intestinal microcirculation following intestinal IR. Six groups of Lewis rats (n = 44) were studied: control, NECA (5'-N-ethyl carboxamide adenosine), IR (30 minutes of intestinal ischemia, 2 hours of reperfusion), IR + NECA, IR + NECA + MRS1754 (A(2B)AR antagonist), IR + NECA + DPCPX (A(1)AR antagonist). All substances were administered i.v. immediately after declamping of the superior mesenteric artery. Intravital microscopy was performed after 2 hours of reperfusion. Following IR we observed a significant increase of leukocyte adhesion in the intestinal submucosal venules and a reduced capillary perfusion within the muscular layers. NECA reduced leukocyte activation and improved capillary perfusion significantly. Administration of A(2B)AR antagonist completely reversed the NECA effect, whereas A(1)AR inhibition only partially abolished the action of NECA. The data support the hypothesis that adenosine signalling is involved in intestinal IR injury. A(2B)AR may be more important than A(1)AR because A(2B)AR inhibition by MRS1754 completely reversed the effect of the adenosine receptor agonist NECA.

• MeSH
• krysa rodu rattus MeSH
• mikrocirkulace účinky léků   MeSH
• modely nemocí na zvířatech MeSH
• potkani inbrední LEW MeSH
• purinergní receptory P1 genetika   metabolismus   MeSH
• reperfuzní poškození MeSH
• střeva krevní zásobení   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Adenosine plays an important role during inflammation, particularly through modulation of monocyte function. The objective of the present study was to evaluate the effect of synthetic adenosine analogs on cytokine production by porcine monocytes. The LPS-stimulated cytokine production was measured by flow cytometry and quantitative real-time PCR. Adenosine receptor expression was measured by quantitative real-time PCR. The present study demonstrates that adenosine analog N-ethylcarboxyamidoadenosine (NECA) down-regulates TNF-α production and up-regulates IL-8 production by LPS-stimulated porcine monocytes. The effect was more pronounced in CD163(-) subset of monocytes compared to the CD163(+) subset. Although both monocyte subsets express mRNA for A1, A2A, A2B and A3 adenosine receptors, the treatment of monocytes with various adenosine receptor agonists and antagonists proved that the effect of adenosine is mediated preferentially via A2A adenosine receptor. Moreover, the study suggests that the effect of NECA on porcine monocytes alters the levels of the cytokines which could play a role in the differentiation of naive T cells into Th17 cells. The results suggest that adenosine plays an important role in modulation of cytokine production by porcine monocytes.

• MeSH
• adenosin-5'-(N-ethylkarboxamid) farmakologie   MeSH
• adenosin farmakologie   MeSH
• agonisté purinergního receptoru P1 farmakologie   MeSH
• antagonisté purinergního receptoru P1 farmakologie   MeSH
• antigeny diferenciační myelomonocytární metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• cytokiny biosyntéza   genetika   MeSH
• lipopolysacharidy farmakologie   MeSH
• messenger RNA genetika   metabolismus   MeSH
• monocyty účinky léků   metabolismus   MeSH
• purinergní receptory P1 genetika   metabolismus   MeSH
• receptory buněčného povrchu metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• Sus scrofa metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The present studies investigated changes in expression of mRNA for adenosine A1, A2a, A2b, and A3 receptors in samples of HL-60 promyelocytic cells differing in the actual presence of cells in various phases of the cell cycle induced by the double thymidine block method. Real-time PCR technique was used for obtaining data on mRNA expression. Statistical analysis of the data revealed that the mRNA expression of adenosine A1, A2a, and A3 receptors is dependent on the cell cycle phase. G0/G1 and G2/M phases were characterized by a higher mRNA expression of adenosine A1 receptors and a lower one of adenosine A2a and A3 receptors whereas the opposite was true for the S phase. Interestingly, expression of mRNA of the adenosine A2b receptors was independent on the cell cycle phase. The results indicate the plasticity of mRNA expression of adenosine receptors in the investigated promyelocytic cells and its interaction with physiological mechanisms of the cell cycle.

• MeSH
• buněčný cyklus genetika   MeSH
• HL-60 buňky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• purinergní receptory P1 genetika   metabolismus   MeSH
• receptor adenosinový A1 genetika   metabolismus   MeSH
• receptor adenosinový A2A genetika   metabolismus   MeSH
• receptor adenosinový A2B genetika   metabolismus   MeSH
• receptor adenosinový A3 genetika   metabolismus   MeSH
• S fáze MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Four mouse bone marrow or thymus cell populations, namely granulopoietic/monocytopoietic, erythropoietic, B-lymphopoietic, and T-lymphopoietic precursor cells have been assayed by RT-PCR technique for the presence and relative amounts of adenosine A(1), A(2a), A(2b), and A(3) receptor mRNA. It has been found that (i) all four populations studied express all four adenosine receptor subtypes, (ii) the A(1), receptor is the least expressed in all populations studied, (iii) the A(3) receptor is markedly expressed in the populations of granulopoietic/monocytopoietic and erythropoietic cells, (iv) the A(2a) receptor is markedly expressed in the populations of B-lymphopoietic and T-lymphopoietic cells, and v) the A(2b) receptor does not predominate in any of the precursor cells studied. Our data offer a new possibility for the assessment of the readiness of these cells to respond, by receptor-mediated mechanisms, to adenosine or its analogs present in the tissues as a result of endogenous processes and/or following their administration.

• MeSH
• financování organizované MeSH
• hematopoetické kmenové buňky metabolismus   MeSH
• hematopoéza genetika   MeSH
• messenger RNA metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• polymerázová řetězová reakce MeSH
• purinergní receptory P1 genetika   MeSH
• receptor adenosinový A1 genetika   MeSH
• receptor adenosinový A2A genetika   MeSH
• receptor adenosinový A2B genetika   MeSH
• receptor adenosinový A3 genetika   MeSH
• regulace genové exprese MeSH
• separace buněk MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH

Expression of mRNA for adenosine receptor subtypes A(1), A(2a), A(2b), and A(3) in normal and lipopolysaccharide (LPS)-activated murine RAW 264.7 macrophages has been investigated using the method of quantitative real-time polymerase chain reaction. The results have shown a very low, unquantifiable expression of adenosine A(1) receptor mRNA in both normal and LPS-activated macrophages. The other three adenosine receptor mRNAs have been found to be expressed at various but always quantifiable levels. Activation of the macrophages by LPS induced upregulation of the expression of adenosine receptor A(2a) and A(2b) mRNA, whereas the expression of adenosine receptor A(3) mRNA was downregulated. Unstimulated macrophages exhibited a high expression of the A(2b) adenosine receptor mRNA. The findings are discussed from the point of view of the antiinflammatory and hematopoiesis-stimulating roles of the adenosine receptor signaling.

• MeSH
• aktivace makrofágů genetika   účinky léků   MeSH
• buněčné linie MeSH
• časové faktory MeSH
• financování organizované MeSH
• hematopoéza genetika   účinky léků   MeSH
• lipopolysacharidy farmakologie   MeSH
• makrofágy metabolismus   účinky léků   MeSH
• messenger RNA metabolismus   MeSH
• myši MeSH
• polymerázová řetězová reakce MeSH
• purinergní receptory P1 genetika   účinky léků   MeSH
• receptor adenosinový A1 genetika   MeSH
• receptor adenosinový A2A genetika   MeSH
• receptor adenosinový A2B genetika   MeSH
• receptor adenosinový A3 genetika   MeSH
• regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH

Cell cycle proteins regulate the transitions from G1 to S and G2 to M phases. In higher eukaryotes, their function is controlled by intracellular cascades regulated by extracellular growth factors. We have studied in previously described transgenic mouse models for thyroid proliferative diseases the expression of the key proteins regulating the cell cycle by Western blotting and immunohistochemistry, and have correlated the observations with the known actions of the transgenes on the signal transduction cascades. In the adenosine A2a receptor model, the cyclic AMP pathway, upstream of the Rb family cell division block, is constitutively activated. In the model expressing HPV 16 E7 protein, the Rb-like proteins are inhibited. Cyclin-dependent kinases cdk4, cdk2 and cdc2, and the associated cyclins D, E and A have been studied. Cyclin D3 appears as the major cyclin D subtype expressed in mouse thyroid epithelial cells in normal and transgenic mice. In the adenosine A2aR model, all cell cycle proteins tested were accumulated. In the E7 model, all cell cycle proteins except for D-type cyclins and cdk4 were also accumulated. A similar pattern was observed in thyroids coexpressing both transgenes, suggesting a dominant effect of E7 over the consequences of the cAMP cascade activation. The cyclin-dependent kinase inhibitors p21cip1/waf1 and p27kip1 were not downregulated in these proliferating thyroids which suggest other roles than the inhibition of the cell cycle progression.

• MeSH
• buněčná diferenciace MeSH
• buněčný cyklus MeSH
• cyklin A metabolismus   MeSH
• cyklin D MeSH
• cyklin E metabolismus   MeSH
• cyklin-dependentní kinasa 2 MeSH
• cyklin-dependentní kinasa 4 MeSH
• cyklin-dependentní kinasy metabolismus   MeSH
• cykliny metabolismus   MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• inhibitor p27 cyklin-dependentní kinasy MeSH
• inhibitory enzymů metabolismus   MeSH
• kinasy CDC2-CDC28 * MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• myši transgenní MeSH
• myši MeSH
• nádorové supresorové proteiny * MeSH
• nádory štítné žlázy metabolismus   MeSH
• onkogenní proteiny virové genetika   metabolismus   MeSH
• Papillomavirus E7 - proteiny MeSH
• protein-serin-threoninkinasy metabolismus   MeSH
• proteinkinasa CDC2 metabolismus   MeSH
• proteiny asociované s mikrotubuly metabolismus   MeSH
• proteiny buněčného cyklu metabolismus   MeSH
• proteiny metabolismus   MeSH
• protoonkogenní proteiny * MeSH
• purinergní receptory P1 genetika   metabolismus   MeSH
• receptor adenosinový A2A MeSH
• štítná žláza metabolismus   MeSH
• thyreoglobulin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH