• MeSH
• RNA virová MeSH

Východisko. Molekulárně biologické metody založené na reverzní transkripci a následné polymerázové řetězové reakci (RT-PCR) jsou schopny detekovat přítomnost transkriptu BCR-ABL u chronické myeloidní leukémie (CML). V tomto sdělení prezentujeme naše zkušenosti se sledováním reziduální leukémie metodou real-time PCR s detekcí hybridizačními sondami u nemocných s CML léčených imatinibem a při sběru periferních krvetvorných buněk (PKB). Metody a výsledky. Stanovení hladin transkriptu BCR-ABL v periferní krvi bylo provedeno u 27 osob před nasazením a v průběhu léčby imatinibem mesylátem. Medián relativního množství BCR-ABL v periferní krvi před nasazením imatinibu byl 2,55 %. Množství transkriptu u 23 osob bez rezistence vůči imatinibu pokleslo během prvních šesti měsíců léčby na hodnotu 0,02 %. Po dvanáctiměsíční léčbě poklesl medián množství transkriptu na 0,005 %. V dalším průběhu léčby hladiny BCR-ABL kolísaly mezi hodnotami pod detekčním limitem metody (0,001 %) a 0,005 %. Tři pacienti byli vůči imatinibu primárně rezistentní s rozsahem hodnot od 0,13 % do 11,7 %. Jeden pacient vykazoval po 18 měsících léčby známky molekulárního relapsu. Z 16 pacientů po stimulaci PKB filgrastimem měly pouze dvě vyšetřované osoby hodnoty BCR-ABL v periferní krvi i v PKB pod detekčním limitem použité metody. U ostatních osob se koncentrace fúzního transkriptu pohybovala v koncentracích spolehlivě stanovitelných metodou RT-PCR. Závěry. Vzhledem k svému prognostickému významu je vyšetření BCR-ABL vhodné pro monitorování kinetiky reziduální choroby. Stanovení provedené v koncentrátu PKB jako potenciálním autotransplantátu může kompletovat informace o kvalitě těchto buněk z hlediska proporce BCR-ABL transkriptu a nastínit další možné terapeutické postupy a prognózu pacienta.

Background. Molecular biology methods based on reverse transcription and polymerase chain reaction (RT-PCR) are able to detect the presence of BCR-ABL transcripts in chronic myeloid leukemia (CML). In this study we present our experience with monitoring of residual disease using real-time PCR with hybridization probes detection in patients treated with imatinib mesylate and in collected peripheral blood progenitor cells (PBPC). Methods and Results.We measured the level of BCR-ABL transcripts in peripheral blood cells of 27 subjects before and in the course of the imatinib treatment. The median of relative quantity of BCR-ABL in the blood before imatinib therapy was 2.55%. The number of the transcripts in 23 imatinib-sensitive subjects decreased to 0.02% in 6 months. After 12 months of the treatment the BCR-ABL median was 0.005%. Subsequent levels fluctuated between values below the detection limit (DL, 0.001%) and 0.005%. Three patients were primarily resistant to imatinib with the BCR-ABL range of 0.13%-11,7% during the treatment. One subject showed marks of molecular relapse after 18 months of the treatment. Only two of 16 filgrastim-stimulated patients had BCR-ABL levels in the blood and in collected PBPC below DL. In other subjects BCR-ABL transcripts were determined within the measurable range of RT-PCR. Conclusions. Taking into account prognostic importance, the measurement of BCR-ABL transcripts is an effective approach to monitoring of residual CML kinetics. Evaluation of BCR-ABL levels in collected PBPC can complete information on quality of the cells in potential autotransplants, and choose subsequent therapeutic protocols and patient prognosis.

• MeSH
• bcr-abl fúzní proteiny krev   MeSH
• chemorezistence MeSH
• chronická myeloidní leukemie diagnóza   farmakoterapie   genetika   MeSH
• faktor stimulující kolonie granulocytů aplikace a dávkování   MeSH
• imatinib mesylát MeSH
• lidé MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   MeSH
• prognóza MeSH
• protinádorové látky aplikace a dávkování   farmakologie   MeSH
• reziduální nádor diagnóza   farmakoterapie   genetika   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

The molecular basis of increased hemoglobin in Andean Aymara highlanders is unknown. We conducted an integrative analysis of whole-genome-sequencing and granulocytes transcriptomics from Aymara and Europeans in Bolivia to explore genetic basis of the Aymara high hemoglobin. Differentially expressed and spliced genes in Aymaras were associated with inflammatory and hypoxia-related pathways. We identified transcripts with 4th or 5th exon skipping of NFKB1 (AS-NFKB1), key part of NF-kB complex, and their splicing quantitative trait loci; these were increased in Aymaras. AS-NFKB1 transcripts correlated with both transcripts and protein levels of inflammatory and HIF-regulated genes, including hemoglobin. While overexpression of the AS-NFKB1 variant led to increased expression of inflammatory and HIF-targeted genes; under inflammatory stress, NF-kB protein translocation to the nucleus was attenuated, resulting in reduced expression of these genes. Our study reveals AS-NFKB1 splicing events correlating with increased hemoglobin in Aymara and their possible protective mechanisms against excessive inflammation.

• MeSH
• alternativní sestřih * genetika   MeSH
• dospělí MeSH
• exony genetika   MeSH
• faktor 1 indukovatelný hypoxií - podjednotka alfa genetika   metabolismus   MeSH
• granulocyty metabolismus   MeSH
• hemoglobiny * metabolismus   genetika   MeSH
• lidé MeSH
• lokus kvantitativního znaku MeSH
• NF-kappa B - podjednotka p50 * metabolismus   genetika   MeSH
• regulace genové exprese MeSH
• transkriptom MeSH
• zánět * genetika   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Bolívie MeSH

BACKGROUND: The programmed cell death 2 (Pdcd2) gene on mouse chromosome 17 was evaluated as a member of a highly conserved synteny, a candidate for an imprinted locus, and a candidate for the Hybrid sterility 1 (Hst1) gene. RESULTS: New mouse transcripts were identified at this locus: an alternative Pdcd2 mRNA skipping the last two coding exons and two classes of antisense RNAs. One class of the antisense RNA overlaps the alternative exon and the other the entire Pdcd2 gene. The antisense RNAs are alternative transcripts of the neighboring TATA-binding protein gene (Tbp) that are located mainly in the cell nucleus. Analogous alternative PDCD2 forms truncating the C-terminal domain were also detected in human and chicken. Alternative transcripts of the chicken PDCD2 and TBP genes also overlap. No correlation in the transcription of the alternative and overlapping mRNAs was detected. Allelic sequencing and transcription studies did not reveal any support for the candidacy of Pdcd2 for Hst1. No correlated expression of Pdcd2 with the other two genes of the highly conserved synteny was observed. Pdcd2, Chd1, and four other genes from this region were not imprinted in the embryo. CONCLUSION: The conservation of alternative transcription of the Pdcd2 gene in mouse, human and chicken suggests the biological importance of such truncated protein. The biological function of the alternative PDCD2 is likely to be opposite to that of the constitutive form. The ratio of the constitutive and alternative Pdcd2 mRNAs differs in the tissues, suggesting a developmental role.The identified Tbp-alternative Pdcd2-antisense transcripts may interfere with the transcription of the Pdcd2 gene, as they are transcribed at a comparable level. The conservation of the Pdcd2/Tbp sense-antisense overlap in the mouse and chicken points out its biological relevance. Our results also suggest that some cDNAs in databases labeled as noncoding are incomplete alternative cDNAs of neighboring protein-coding genes.

• MeSH
• alely MeSH
• alternativní sestřih MeSH
• apoptóza imunologie   MeSH
• exprimované sekvenční adresy MeSH
• financování organizované MeSH
• genomový imprinting MeSH
• krysa rodu rattus MeSH
• kur domácí MeSH
• lidé MeSH
• mapování chromozomů MeSH
• messenger RNA genetika   MeSH
• molekulární sekvence - údaje MeSH
• myši MeSH
• polymerázová řetězová reakce MeSH
• proteiny regulující apoptózu genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH

• MeSH
• biologické markery MeSH
• biopsie MeSH
• dysplazie děložního hrdla diagnóza   genetika   patologie   diagnóza   genetika   patologie   MeSH
• exprese genu MeSH
• financování organizované MeSH
• genetická transkripce MeSH
• imunohistochemie metody   MeSH
• infekce papilomavirem diagnóza   genetika   MeSH
• inhibitor p16 cyklin-dependentní kinasy analýza   genetika   MeSH
• lidé MeSH
• lidský papilomavirus 16 MeSH
• lidský papilomavirus 18 MeSH
• messenger RNA genetika   MeSH
• nádory děložního čípku diagnóza   genetika   patologie   MeSH
• pilotní projekty MeSH
• plocha pod křivkou MeSH
• polymerázová řetězová reakce MeSH
• prospektivní studie MeSH
• senzitivita a specificita MeSH
• statistika jako téma MeSH
• vaginální stěr MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• hodnotící studie MeSH
• srovnávací studie MeSH

PURPOSE: Tumour cells killing by cytotoxic therapies largely depends on triggering the intrinsic apoptosome-mediated caspase activation pathway but it had never been evaluated whether the expression of transcripts encoding the core components of apoptosome pathway is altered in non-small cell lung carcinoma (NSCLC). METHODS: We investigated the expression status of several apoptosome pathway-related transcripts including Apaf-1, procaspase-9, -3, -6, -7 and Smac in tumour and lung tissue samples from 65 surgically treated NSCLC patients and in 10 NSCLC cell lines with using real time RT-PCR. RESULTS: NSCLC tissues and cell lines showed significantly increased expression of procaspase-9, -3, -6 and Smac mRNAs as compared to the lungs and expression of these transcripts was simultaneously upregulated in a subset of NSCLCs belonging to different histopathological type, grade and stage categories. The expression of procaspase-7 mRNA in NSCLC tissues and cell lines and lungs was not significantly different. By contrast, the expression of Apaf-1 mRNA was frequently downregulated in the tumours as compared to matched lungs. Nevertheless, the examined NSCLC cell lines showed significantly higher expression of Apaf-1 mRNA than the lungs. The expression of Apaf-1, procaspase-9 and -6 mRNAs was higher in lung adenocarcinomas as compared to squamous cell lung carcinomas but the expression levels of the studied apoptosome pathway-related transcripts in the tumours were independent of tumour's grade and stage. CONCLUSIONS: The results of the present study suggest that there is a subgroup of NSCLCs, which may be intrinsically primed for apoptosis through upregulated expression of transcripts encoding the apoptosome pathway components.

• MeSH
• aktiny metabolismus   MeSH
• apoptóza imunologie   MeSH
• dospělí MeSH
• down regulace MeSH
• faktor 1 aktivující apoptotickou proteasu MeSH
• financování organizované MeSH
• genetická transkripce MeSH
• intracelulární signální peptidy a proteiny genetika   metabolismus   MeSH
• kaspasa 3 MeSH
• kaspasa 6 MeSH
• kaspasa 7 MeSH
• kaspasa 9 MeSH
• kaspasy genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé genetika   metabolismus   MeSH
• messenger RNA metabolismus   MeSH
• mitochondriální proteiny genetika   metabolismus   MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• nádory plic genetika   metabolismus   MeSH
• nemalobuněčný karcinom plic genetika   metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prekurzory enzymů metabolismus   MeSH
• proteiny genetika   metabolismus   MeSH
• regulace genové exprese u nádorů MeSH
• senioři MeSH
• upregulace MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé genetika   metabolismus   MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH

Antisense transcripts play an important role in generating regulatory non-coding RNAs but whether these transcripts are also translated to generate functional peptides remains poorly understood. In this study, RNA sequencing and six-frame database generation were combined with mass spectrometry analysis of peptides isolated from polysomes to identify Nascent Pioneer Translation Products (Na-PTPs) originating from alternative reading frames of bi-directional transcripts. Two Na-PTP originating peptides derived from antisense strands stimulated CD8+ T cell proliferation when presented to peripheral blood mononuclear cells (PBMCs) from nine healthy donors. Importantly, an antigenic peptide derived from the reverse strand of two cDNA constructs was presented on MHC-I molecules and induced CD8+ T cell activation. The results demonstrate that three-frame translation of bi-directional transcripts generates antigenic peptide substrates for the immune system. This discovery holds significance for understanding the origin of self-discriminating peptide substrates for the major histocompatibility class I (MHC-I) pathway and for enhancing immune-based therapies against infected or transformed cells.

• MeSH
• aktivace lymfocytů imunologie   MeSH
• antisense RNA * genetika   imunologie   MeSH
• CD8-pozitivní T-lymfocyty * imunologie   MeSH
• leukocyty mononukleární imunologie   MeSH
• lidé MeSH
• MHC antigeny I. třídy * imunologie   genetika   MeSH
• peptidy * imunologie   genetika   MeSH
• prezentace antigenu MeSH
• proteosyntéza * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Článok sa zaoberá výskumným overovaním otázky, či imaginatívna fáza v metóde katatýmne imaginatívnej psychoterapie (KIP) aktivizuje, tak ako sa predpokladá, primárny proces. Sú prezentované výsledky pre účely výskumu vytvorenej textovej analýzy prepisov terapeutických sedení realizovaných metódou katatýmne imaginatívnej psychoterapie. Dôležitým je zistenie významne vyššieho výskytu primárne procesových verbálnych výrazov v imaginatívnej časť KIP sedení. Vtedy sa dostávame prostredníctvom obrazov k nevedomým častiam osobnosti klienta a k obsahom emocionálnej a procedurálnej pamäte, ktoré potom môžu byť v nasledujúcich rozhovorových fázach terapie ďalej oslovované a spracované tak, aby sa mohli začleniť do vedomej časti osobnosti.

The article deals with research verifying of the question whether imaginative stage in Guided Affective Imagery (GAI) activates, as it is assumed, the primary process. There are results presented for the purpose of created text analysis research of therapeutic sessions transcripts realized by the method Guided Affective Psychotherapy. An important finding concerns the significantly higher occurrence of primary processing verbal aspects in the imagery part of the GAI session. At this time, we get, by means of images, to unconscious parts of client's personality and to emotional and procedural memory contents, which then, in following interview stages of therapy, can be further appealed and processed so that they could be integrated into the conscious part of personality.

• MeSH
• imaginativní postupy v psychoterapii metody   statistika a číselné údaje   MeSH
• lidé MeSH
• psychoterapie metody   MeSH
• sémantika MeSH
• statistika jako téma MeSH
• výzkum statistika a číselné údaje   MeSH
• Check Tag
• lidé MeSH

Euglena gracilis possesses secondary plastids of green algal origin. In this study, E. gracilis expressed sequence tags (ESTs) derived from polyA-selected mRNA were searched and several ESTs corresponding to plastid genes were found. PCR experiments failed to detect SL sequence at the 5'-end of any of these transcripts, suggesting plastid origin of these polyadenylated molecules. Quantitative PCR experiments confirmed that polyadenylation of transcripts occurs in the Euglena plastids. Such transcripts have been previously observed in primary plastids of plants and algae as low-abundance intermediates of transcript degradation. Our results suggest that a similar mechanism exists in secondary plastids.

• MeSH
• Euglena gracilis genetika   metabolismus   účinky záření   MeSH
• exprimované sekvenční adresy MeSH
• genom plastidový MeSH
• messenger RNA genetika   metabolismus   MeSH
• plastidy genetika   metabolismus   MeSH
• polyadenylace * MeSH
• protozoální geny MeSH
• RNA protozoální genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Protein p73 is a member of the p53 protein family that can induce cell cycle arrest or apoptosis by the activation of p53-responsive genes as well as p53-independent pathways. Alternative promoter usage, together with differential splicing of the C-terminal exons, forms several distinct mRNAs that are translated into corresponding protein isoforms containing different domains. While TAp73 isoforms respond to genotoxic stress in a manner similar to tumor suppressor p53, ΔTAp73 isoforms inhibit apoptosis during normal development and in cancer cell lines. Thus, the impact of p73 on tumorigenesis depends on a subtle balance between tumor-promoting and -suppressing isoforms. Due to the structural homology between p53 and p73, a subtle balance among p53 family members and their isoforms could influence glioma cell evolution toward malignancy. Thus, the p73 status has to be considered when studying the regulatory role of p53 protein in gliomagenesis. The presented review summarizes recent knowledge about the issue of p73 and its isoforms with respect to neuro-oncology research.

• MeSH
• DNA vazebné proteiny genetika   metabolismus   MeSH
• genetická transkripce MeSH
• jaderné proteiny genetika   metabolismus   MeSH
• lidé MeSH
• mutace genetika   MeSH
• nádorová transformace buněk genetika   metabolismus   MeSH
• nádorové supresorové proteiny genetika   metabolismus   MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• nádory centrálního nervového systému genetika   metabolismus   MeSH
• protein - isoformy genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH