BACKGROUND AND OBJECTIVES: Utilisation of the one-step nucleic acid amplification (OSNA) molecular biology method for the detection of the metastatic involvement of sentinel lymph nodes (SLNs) in endometrial cancer (EC) patients. A comparison with histopathological ultrastaging and a description of the clinical consequences. METHODS: Surgically treated EC patients underwent detection of SLNs. Nodes greater than 5 mm were cut into sections 2-mm thick parallel to the short axis of the node. Odd sections were examined according to the OSNA method, while even ones according to an appropriate ultrastaging protocol. Nodes less than or equal to 5 mm were cut into halves along the longitudinal axis with one half examined according to the OSNA method and the other half by ultrastaging. RESULTS: Fifty-eight patients were included and 135 SLNs were acquired. Both ultrastaging and OSNA agreed on 116 results. According to the OSNA method, 20.69% more patients were classified into International Federation of Gynecology and Obstetrics (FIGO) stage III. When comparing the results of the OSNA method to the conclusions of ultrastaging as a reference method, sensitivity of 90.9%, specificity of 85.5% and concordance of 85.9% were attained. CONCLUSIONS: The results of the OSNA method showed a higher frequency of detection of micrometastases and included 20.69% more patients into FIGO stage III.
- MeSH
- adenokarcinom z jasných buněk genetika sekundární chirurgie MeSH
- biopsie sentinelové lymfatické uzliny MeSH
- dospělí MeSH
- keratin-19 genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- lymfatické metastázy MeSH
- mikrometastázy MeSH
- míra přežití MeSH
- nádorové biomarkery genetika MeSH
- nádory endometria genetika patologie chirurgie MeSH
- následné studie MeSH
- nukleové kyseliny analýza genetika MeSH
- peroperační doba MeSH
- prognóza MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- sentinelová uzlina patologie chirurgie MeSH
- serózní cystadenokarcinom genetika sekundární chirurgie MeSH
- techniky amplifikace nukleových kyselin metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
Článek přináší základní orientaci v dějích a pojmech, které se podílejí na kancerogenezi. Neklade si za cíl přinést vyčerpávající odbornou informaci, ale jednoduchou a srozumitelnou formou předkládá terminologický výklad pojmů, které i začínající onkolog běžně používá. Tyto pojmy je vhodné znát, ale především je třeba jim rozumět. Článek vysvětluje charakteristiky přepisu genetické informace a tvorby bílkovin, zabývá se chromosomálními i genovými mutacemi. Popisuje význam genetických mutací pro vznik karcinomu plic.
The article provides a basic orientation in the processes and concepts involved in carcinogenesis. It does not aim to bring exhaustive expert information, but in a simple and comprehensible form it presents a terminological interpretation of the terms commonly used by a beginning oncologist. These concepts are useful to know, but also to understand them. The article explains the characteristics of transcription of genetic information and protein synthesis, deals with both chromosomal and gene mutations. It describes the importance of genetic mutations for lung cancer.
We have carried out extended set of μs-scale explicit solvent MD simulations of all possible G-triplexes which can participate in folding pathways of the human telomeric quadruplex. Our study accumulates almost 60 μs of simulation data, which is by about three orders of magnitude larger sampling compared to the earlier simulations of human telomeric G-DNA triplexes. Starting structures were obtained from experimental quadruplex structures by deleting either the first or the last strand. The life-times of antiparallel triplexes with lateral and diagonal loops are at least on μs-scale, which should be sufficient to contribute to the folding pathways. However, the triplex states may involve structures with various local deviations from the ideal triplexes, such as strand tilting and various alternative and incomplete triads. The simulations reveal easy rearrangements between lateral and diagonal loop triplex topologies. Propeller loops of antiparallel triplexes may to certain extent interfere with the G-triplexes but these structures are still viable candidates to participate in the folding. In contrast, all-parallel all-anti triplexes are very unstable and are unlikely to contribute to the folding. Although our simulations demonstrate that antiparallel G-triplexes, if folded, would have life-times sufficient to participate in the quadruplex folding, the results do not rule out the possibility that the G-triplexes are out-competed by other structures not included in our study. Among them, numerous possible misfolded structures containing guanine quartets can act as off-path intermediates with longer life-times than the triplexes. Besides analyzing the structural dynamics of a diverse set of G-DNA triplexes, we also provide a brief discussion of the limitations of the simulation methodology, which is necessary for proper understanding of the simulation data.
- MeSH
- DNA chemie MeSH
- G-kvadruplexy * MeSH
- konformace nukleové kyseliny * MeSH
- lidé MeSH
- nukleové kyseliny chemie genetika MeSH
- sekvence nukleotidů MeSH
- simulace molekulární dynamiky MeSH
- telomery chemie genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Based on the matrix-addressing sequence of mitochondrial ribosomal 5S-rRNA (termed MAM), which is naturally imported into mitochondria, we have constructed an import system for in vivo targeting of mitochondrial DNA (mtDNA) or mt-mRNA, in order to provide fluorescence hybridization of the desired sequences. Thus DNA oligonucleotides were constructed, containing the 5'-flanked T7 RNA polymerase promoter. After in vitro transcription and fluorescent labeling with Alexa Fluor(®) 488 or 647 dye, we obtained the fluorescent "L-ND5 probe" containing MAM and exemplar cargo, i.e., annealing sequence to a short portion of ND5 mRNA and to the light-strand mtDNA complementary to the heavy strand nd5 mt gene (5'-end 21 base pair sequence). For mitochondrial in vivo fluorescent hybridization, HepG2 cells were treated with dequalinium micelles, containing the fluorescent probes, bringing the probes proximally to the mitochondrial outer membrane and to the natural import system. A verification of import into the mitochondrial matrix of cultured HepG2 cells was provided by confocal microscopy colocalizations. Transfections using lipofectamine or probes without 5S-rRNA addressing MAM sequence or with MAM only were ineffective. Alternatively, the same DNA oligonucleotides with 5'-CACC overhang (substituting T7 promoter) were transcribed from the tetracycline-inducible pENTRH1/TO vector in human embryonic kidney T-REx®-293 cells, while mitochondrial matrix localization after import of the resulting unlabeled RNA was detected by PCR. The MAM-containing probe was then enriched by three-order of magnitude over the natural ND5 mRNA in the mitochondrial matrix. In conclusion, we present a proof-of-principle for mitochondrial in vivo hybridization and mitochondrial nucleic acid import.
- MeSH
- genetická transkripce MeSH
- lidé MeSH
- mitochondriální DNA chemie genetika MeSH
- nukleové kyseliny chemie genetika MeSH
- oligonukleotidy chemie genetika MeSH
- RNA ribozomální chemie genetika MeSH
- RNA chemie genetika MeSH
- sekvenční homologie nukleových kyselin MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- celotělové ozáření škodlivé účinky MeSH
- finanční podpora výzkumu jako téma MeSH
- hematopoetický systém účinky léků účinky záření MeSH
- játra účinky léků účinky záření MeSH
- kadmium aplikace a dávkování škodlivé účinky MeSH
- krysa rodu rattus MeSH
- ledviny účinky léků účinky záření MeSH
- nukleové kyseliny genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- chromatin ultrastruktura MeSH
- genetická transkripce MeSH
- genetický kód MeSH
- nukleové kyseliny genetika MeSH
- Publikační typ
- přehledy MeSH
