The purpose of our study was to examine an early activation of JNK and p38 mitogen activated protein kinases (MAPK) and their substrate c-Myc after soman poisoning in order to enlighten the pathogenetic mechanism of nerve agent-induced non-specific effects. Male Wistar rats were intramuscularly poisoned by soman (60 μg.kg-1 - 70% LD50). Samples were taken 4, 24, and 72 hours after poisoning, immunohistochemically stained and phospho-JNKThr-183/Tyr-185, phospho-p38Thr180/Tyr182, and phospho-c- MycThr58/Ser62 expressions were measured using a computer Image analysis in apical and cryptal enterocytes of the colon transversum. We observed decreased phospho-JNK in apical enterocytes 4 and 24 h after poisoning and increased phospho-JNK in cryptal and apical enterocytes 72 h after intoxication. Phosphop38 dropped significantly in the apical compartment 72 h after soman poisoning. An activation of c-Myc decreased in both apical and cryptal compartment 4 and 24 h after soman intoxication, while increased in both compartments 72 h after poisoning. Soman poisoning seems to temporarily suppress promitotic pathways of proliferating cryptal cells and causes delayed activation of JNK stress signaling pathway.
- MeSH
- časové faktory MeSH
- colon transversum chemie MeSH
- enterocyty chemie MeSH
- experimenty na zvířatech MeSH
- financování organizované MeSH
- JNK mitogenem aktivované proteinkinasy chemie MeSH
- kontrolní skupiny MeSH
- MAP kinasový signální systém fyziologie MeSH
- mitogenem aktivované proteinkinasy p38 chemie MeSH
- potkani Wistar MeSH
- protoonkogenní proteiny c-myc chemie MeSH
- soman otrava toxicita MeSH
- výzkumný projekt MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
The purpose of our study was to examine the early expression of p21 and activated transcription factors ATF-2, CREB, Elk-1, p53 after soman and VX poisoning, to throw light on the pathogenetic mechanism of nerve agent-induced non-specific effects. Male Wistar rats were i.m. poisoned by soman (60 μg.kg-1 – 70% LD50) or VX (8 μg.kg-1 – 70% LD50). Samples were taken 4, 24, and 72 hours after poisoning, immunohistochemically stained and phospho-ATF-2Thr-69/71, phospho-CREBSer-133, phospho-Elk-1Ser-383, phospho-p53Ser-15, and protein p21 expressions were measured using computer Image analysis in apical and cryptal enterocytes of the colon transversum. After soman poisoning, we observed an increased phospho-CREB in cryptal enterocytes 4, 24, and 72 h after poisoning, while apical enterocytes expressed increased phospho-CREB only 72 h after intoxication. Phospho-Elk-1 significantly dropped 4 and 24 h after soman poisoning in the cryptal compartment. Activation of ATF-2 and p53 and expression of p21 were not changed 4, 24, and 72 h after soman poisoning. VX poisoning did not change any of measured parameters. Soman and VX showed a different effect on cellular signalling. Soman seems to cause additional effects, which are not related to the basic mechanism of nerve agent-induced toxicity and which temporarily suppress promitotic pathways of proliferating cells and persist in cells during the differentiation process.
- Klíčová slova
- VX,
- MeSH
- aktivační transkripční faktory účinky léků MeSH
- enterocyty účinky léků MeSH
- financování organizované MeSH
- histologické techniky metody statistika a číselné údaje MeSH
- nádorový supresorový protein p53 účinky léků MeSH
- onkogenní protein p21(ras) účinky léků MeSH
- organofosforové sloučeniny škodlivé účinky MeSH
- počítačové zpracování obrazu statistika a číselné údaje MeSH
- potkani Wistar MeSH
- protein Elk-1 s doménou ets účinky léků MeSH
- protein vázající CREB účinky léků MeSH
- soman škodlivé účinky MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
PURPOSE: To examine the p38 mitogen-activated protein kinase (p38) phosphorylation and transforming growth factor beta 1 (TGF-β1) expression in rat colon enterocytes after irradiation and their contribution to pathology of intestinal radiation disease. MATERIALS AND METHODS: Male Wistar rats were irradiated with whole body γ-radiation of 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 Gy ((60)Co, 1.44 Gy.min(-1)). Samples were taken 4 and 24 h after irradiation, immunohistochemically stained, then p38 phosphorylation and TGF-β1 expression were measured in apical and cryptal enterocytes using computer image analysis. In selected groups, morphometric parameters, mitosis and apoptosis were evaluated. RESULTS: P38 phosphorylation integrated optical density (IOD)-based levels increased 2.4-fold (p ≤ 0.01) and 3.6 to 22.8-fold (p ≤ 0.001) in apical enterocytes 4 h after 0.5 Gy and 24 h after 3-10 Gy, respectively. TGF-β1 IOD-based expression increased 3.3- to 6.9-fold (p ≤ 0.001) and 1.6- to 4.9-fold (p ≤ 0.001) in apical cells 4 h after 0.5-2, 4, 5 Gy and 24 h after 6-10 Gy, respectively. No changes were observed in crypts. CONCLUSIONS: We found a chronological and dose-dependent order of p38 activation and TGF-β1 expression in apical enterocytes. Transient up-regulation of p38 and TGF-β1 signalling observed 4 h after low-dose irradiation may participate in molecular mechanisms creating cellular over-expression in apical compartment, while persistent patterns measured 24 h after high-dose irradiation might provide protection of remaining cells in order to maintain tissue integrity.
- MeSH
- aktivace enzymů účinky záření MeSH
- apoptóza účinky záření MeSH
- časové faktory MeSH
- celotělové ozáření škodlivé účinky MeSH
- enterocyty cytologie enzymologie metabolismus účinky záření MeSH
- fosforylace účinky záření MeSH
- kolon cytologie účinky záření MeSH
- krysa rodu rattus MeSH
- mitogenem aktivované proteinkinasy p38 metabolismus MeSH
- mitóza účinky záření MeSH
- polarita buněk účinky záření MeSH
- potkani Wistar MeSH
- regulace genové exprese účinky záření MeSH
- záření gama škodlivé účinky MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- lékařské mise MeSH
- ozbrojené síly MeSH
- radiobiologie MeSH
- vojenské lékařství MeSH
- vojenské nemocnice MeSH
- Publikační typ
- novinové články MeSH
PURPOSE: The large white pig is a useful experimental model to compare in vivo, in vitro and ex vivo sensitivity of peripheral blood leukocytes to ionising radiation. Such studies are impossible to perform in humans and laboratory rodents due to ethical reasons and body size, respectively. We analysed dose- and time-dependent changes of lymphocyte and granulocyte absolute numbers in porcine peripheral blood after either whole-body irradiation (in vivo and ex vivo experiments) or exposure of porcine whole blood to γ-irradiation (in vitro experiments). MATERIALS AND METHODS: CytoCount™ absolute counting beads and light scatter analysis using a flow cytometer were used to determine major leukocyte subpopulation numbers in blood samples after red cell removal. RESULTS: Similar to other species, lymphocyte numbers significantly decreased in pigs both in vivo and in vitro in a dose-dependent manner. Most importantly, our data clearly show that reduction of lymphocyte numbers after irradiation in vivo proceeds much faster than after irradiation in vitro and that granulocyte changes depend only on the time of analysis after irradiation. CONCLUSIONS: All three tested experimental arrangements demonstrated the radiosensitivity of lymphocytes and the radioresistance of peripheral blood granulocytes. These in vivo and in vitro approaches, as well as the newly introduced ex vivo observations, appear to be relevant to biodosimetry.
- MeSH
- celotělové ozáření MeSH
- ionizující záření MeSH
- leukocyty cytologie účinky záření MeSH
- počet buněk metody MeSH
- prasata MeSH
- průtoková cytometrie metody MeSH
- radiační rozptyl MeSH
- světlo MeSH
- techniky in vitro MeSH
- tolerance záření MeSH
- vztah dávky záření a odpovědi MeSH
- záření gama MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- biodozimetrie,
- MeSH
- dávka záření MeSH
- diagnostické techniky molekulární metody využití MeSH
- financování organizované MeSH
- ionizující záření MeSH
- krevní obraz klasifikace metody využití MeSH
- lidé MeSH
- monitorování radiace metody normy přístrojové vybavení MeSH
- příznaky a symptomy MeSH
- radiační poranění diagnóza patofyziologie MeSH
- radiobiologie metody normy MeSH
- radiometrie metody využití MeSH
- statistika jako téma MeSH
- Check Tag
- lidé MeSH
AIM: The aim of this study is to evaluate the diagnostic yield of capsule endoscopy in nonsteroidal anti-inflammatory drug (NSAID)-induced enteropathy in pigs. MATERIALS AND METHODS: Indomethacin (400 mg/day) was administrated orally for 10 days to eight female pigs weighing 36.3+/-2.4 kg. Afterwards, capsule endoscopy was performed, using the EndoCapsule system (Olympus Optical Co., Tokyo, Japan). The following morning, pharmacological euthanasia and immediate autopsy were performed. RESULTS: Small bowel injury compatible with NSAID-induced enteropathy was observed in 7/8 animals. The most common lesions were red spots and erosions. Ulcers and small intestinal bleeding were identified sporadically. Sensitivity and specificity of capsule endoscopy were 83.3% and 95.8%, respectively. CONCLUSION: Our results indicate that wireless capsule endoscopy is a highly accurate noninvasive method for evaluation of experimental NSAID-induced enteropathy.
- MeSH
- antiflogistika nesteroidní MeSH
- duodenum patologie MeSH
- dvanáctníkové vředy chemicky indukované diagnóza MeSH
- ileum patologie MeSH
- indomethacin MeSH
- jejunum patologie MeSH
- kapslová endoskopie MeSH
- krvácení z peptického vředu chemicky indukované diagnóza MeSH
- modely nemocí na zvířatech MeSH
- nemoci střev chemicky indukované diagnóza MeSH
- prediktivní hodnota testů MeSH
- senzitivita a specificita MeSH
- střevní sliznice patologie MeSH
- Sus scrofa MeSH
- tenké střevo patologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
In this work we evaluated changes in molecular response of human promyelocyte leukemia cells HL-60 and HL-60-IR cells (HL-60 irradiated by 10 cycles of radiation with total dose of 60 Gy, given over a period of 3 months) to irradiation by the dose of 2 and 8 Gy. Analysis of CD11b and apoptosis by flow-cytometry revealed that on 3rd day after irradiation by 8 Gy the HL-60-IR are more resistant to radiation-induced apoptosis and more differentiated (increase in CD11b in non-apoptotic cells) than regular HL-60. We found that both types of cells have high basal level of phosphorylated extracellular signal-regulated kinases Erk1/2 . Irradiation induces decrease in Erk1/2 phosphorylation after 4 and 8 h in both cell types. However, in HL-60-IR cells Erk1/2 phosphorylation is restored faster than in HL-60. Also it was found that in contrary to HL-60 cells, the HL-60-IR cells react to 2 Gy irradiation by p53 independent increase in p21(WAF1/Cip1), and not by activation of checkpoint kinase Chk-2. Therefore we concluded that relatively high dose of radiation (6 Gy) does not lead after 10 repetitive irradiations to eradication of HL-60 cells, but instead increases their radioresistance, increases the ability to differentiate, alters MAPK response, increases amount of p21(WAF1/Cip1), and decreases induction of apoptosis by ionizing radiation. p21(WAF1/Cip1) might prevent apoptosis induction and trigger permanent cell-cycle arrest, which can also contribute to regression of this leukaemia after therapy.
- MeSH
- akutní promyelocytární leukemie metabolismus patologie radioterapie MeSH
- apoptóza účinky záření MeSH
- buněčná diferenciace účinky záření MeSH
- buněčný cyklus účinky záření MeSH
- fosforylace MeSH
- frakcionace dávky záření MeSH
- HL-60 buňky MeSH
- inhibitor p21 cyklin-dependentní kinasy metabolismus MeSH
- lidé MeSH
- MAP kinasový signální systém účinky záření MeSH
- tolerance záření fyziologie MeSH
- vztah dávky záření a odpovědi MeSH
- záření gama terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
