• Publikační typ
• abstrakt z konference MeSH

The tumor suppressor protein p53 is a key factor in genome stability and one of the most studied of DNA binding proteins. This is the first study on the interaction of wild-type p53 with guanine quadruplexes formed by the human telomere sequence. Using electromobility shift assay and ELISA, we show that p53 binding to telomeric G-quadruplexes increases with the number of telomeric repeats. Further, p53 strongly favors G-quadruplexes folded in potassium over those formed in sodium, thus indicating the telomeric G-quadruplex conformational selectivity of p53. The presence of the quadruplex-stabilizing ligand, N-methyl mesoporphyrin IX (NMM), increases p53 recognition of G-quadruplexes in potassium. Using deletion mutants and selective p53 core domain oxidation, both p53 DNA binding domains are shown to be crucial for telomeric G-quadruplex recognition.

• MeSH
• cirkulární dichroismus MeSH
• DNA chemie   genetika   metabolismus   MeSH
• draslík chemie   MeSH
• ELISA MeSH
• G-kvadruplexy * MeSH
• kompetitivní vazba MeSH
• lidé MeSH
• mesoporfyriny chemie   MeSH
• mutace MeSH
• nádorový supresorový protein p53 chemie   genetika   metabolismus   MeSH
• oligonukleotidy chemie   genetika   metabolismus   MeSH
• retardační test MeSH
• sekvence nukleotidů MeSH
• tandemové repetitivní sekvence genetika   MeSH
• telomery chemie   genetika   metabolismus   MeSH
• vazba proteinů MeSH
• vazebná místa genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Úspešnosť liečby Ta-T1 nádorov močového mechúra (NMM) závisí od precízne realizovanej transuretrálnej resekcie (TURBT) všetkých papilárnych nádorov, výberu adekvátnej adjuvantnej intravezikálnej liečby a správneho sledovania pacientov s týmto ochorením. Vysoká početnosť recidív je dôkazom nedostatočnej a nesprávnej terapie NMM s podcenením rizika pre pacienta. Ani kvalitne uskutočnená TURBT nezabráni vzniku recidív (s frekvenciou 50-70 %), resp. progresii ochorenia (s výskytom do 30 %) v závislosti od klinických a histopatologických vlastností papilárnych nádorov (patologické štádium, grading, počet a veľkosť nádorov, či ide o primárny, alebo už recidivujúci nádor a pod.). Ďalšia, intravezikálna, liečba je preto nevyhnutným predpokladom zníženia rizika recidívy alebo progresie. Po TURBT je u pacientov s nízkym rizikom recidívy a progresie indikovaná jednorazová instilácia chemoterapeutika do 6 hodín po zákroku. U pacientov so stredným a s vysokým rizikom recidívy je jednorazová instilácia chemoterapeutika suboptimálna, preto pokračujeme v adjuvantnej liečbe. Intravezikálna imunoterapia je pravdepodobne účinnejšia ako chemoterapia, avšak jej lepšia účinnosť bola preukázaná až pri aplikovaní najmenej 1-ročnej udržiavacej liečby. Často je však táto liečba v klinickej praxi ukončená už po indukčnom cykle (výskyt nežiaducich účinkov, nespolupráca pacienta, rozhodnutie lekára), pacienti zostávajú ďalej neliečení a výsledkom je neúspech v podobe recidívy alebo progresie ochorenia. Pri strednom riziku recidívy a nízkom riziku progresie je určite rovnocennou alternatívou intravezikálna chemoterapia. Ak existuje predpoklad, že u pacienta s vysokým rizikom recidívy sa nebude dať aplikovať najmenej 1-ročná intravezikálna imunoterapia podľa odporúčanej schémy, takisto možno indikovať intravezikálnu chemoterapiu, ako pacienta liečiť imunoterapiou v suboptimálnom režime alebo ho dokonca neliečiť vôbec, čo, bohužiaľ, nie je zriedkavé. Nádory močového mechúra sú aj v Ta-T1 štádiu onkologickým ochorením s možným letálnym koncom a prognóza neadekvátne liečených pacientov je závažná.

The fruitfulness of Ta-T1 bladder tumours (NMIBC) therapy depends on a meticulous transurethral resection of all papillary lesions (TURBT), a selection of appropriate adjuvant intravesical therapy and a proper follow-up of patients with such disease. High frequency of recurrence indicates an insufficient and improper therapy of NMIBC with underestimated risk for the patient. Even a superior TURBT cannot prevent a development of recurrence (with frequency 50-70 %) or progression (with up to 30 % rate) depending on clinical and histopathological characteristics of papillary tumours (pathological stage, grade, number and size of tumours, if it is primary or recurrent tumour etc.). Further, intravesical therapy is therefore the essential tool to reduce the risk of recurrence or progression. An immediate single instillation of chemotherapeutic agent within 6 hours after surgery is indicated as a definitive treatment for patients with low risk of recurrence and progression. A single instillation is suboptimal in case of intermediate or high risk of recurrence, therefore adjuvant therapy should continue. Intravesical immunotherapy is probably more effective than chemotherapy; however, its superiority was proven only with maintenance therapy lasting at least 1 year. But this therapy is in real clinical practice often terminated just after induction course (adverse events, non-compliance of patient, urologist decision), patients remain untreated with resulting treatment failure (recurrence or progression of disease). Intravesical chemotherapy is definitely an equal treatment option to immunotherapy in case of an intermediate risk of recurrence and a low risk of progression. If we can assume that the patient with high risk of recurrence will not complete at least 1-year intravesical therapy according to recommended schedule, we should rather prefer intravesical chemotherapy than a suboptimal course of immunotherapy or even no treatment, which unfortunately is not a rare case. Remember, bladder cancer in Ta-T1 stage is still oncological, potentially lethal condition and prognosis of inadequately managed patients is serious.

Cieľom práce bolo vyhodnotiť kvalitu primárnej TURB, stanoviť výskyt reziduálneho nádoru (RN) a vplyv klinických a histopatologických charakteristík primárneho nádoru na riziko ponechania RN. Do retrospektívnej štúdie sme zaradili 260 pacientov s novodiagnostikovaným svalovinu neinfiltrujúcim stredne a vysoko rizikovým NMM, reTURB absolvovali 124 pacienti. Hodnotili sme pohlavie, vek, pT, grading, počet a veľkosť primárnych nádorov, prítomnosť/neprítomnosť svaloviny v resekovanom tkanive, podanie/nepodanie epirubicínu do 6 hodín po TURB. Na štatistické vyhodnotenie boli použité jednofaktorové a multifaktorové analýzy. V našom súbore sme zistili 40% výskyt RN a 2,4% výskyt infiltrácie svaloviny pri reTURB. Prediktormi RN boli stupeň diferenciácie buniek a nepriaznivý cystoskopický nález. Peroperačná instilácia epirubicínu neovplyvnila výskyt RN. Neprítomnosť svaloviny vo vzorkách po primárnej TURB neviedla v našom súbore k vyššej incidencii RN.

The aim of study was to evaluate the quality of primary transurethral resection of bladder tumour, to determine the incidence of residual tumour (RT) and the impact of clinical and histopathological characteristics of primary tumour on the risk of occurence of RT. Consecutive 260 patients with intermediate and high risk primary non-muscle invasive bladder cancer were included in the study, repeat TUR was performed in 124 patients. Gender, age, pT stage, grade, number and diameter of primary tumours, presence/absence of muscle in TUR specimen and intravesical instillation of epirubicin within 6 hours after TUR were evaluated. Statistical univariate and multivariate analysis was performed. In our cohort the incidence of RT was 40 % with 2.4 % incidence of muscle invasion revealed in reTUR specimen. Significant predictors of RT were grade and unfavourable cystoscopic findings. Immediate intravesical epirubicin instillation did not change the incidence of RT. The absence of muscle in primary TUR specimen did not lead to the increased incidence of RT.

• MeSH
• lidé MeSH
• nádory močového měchýře chirurgie   MeSH
• reziduální nádor epidemiologie   MeSH
• staging nádorů MeSH
• urologické chirurgické výkony u mužů metody   MeSH
• urologické chirurgické výkony metody   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

BACKGROUND: Many studies have shown that guanine-rich DNA sequences form quadruplex structures (G4) in vitro but there is scarce evidence of guanine quadruplexes in vivo. The majority of potential quadruplex-forming sequences (PQS) are located in transposable elements (TEs), especially close to promoters within long terminal repeats of plant LTR retrotransposons. RESULTS: In order to test the potential effect of G4s on retrotransposon expression, we cloned the long terminal repeats of selected maize LTR retrotransposons upstream of the lacZ reporter gene and measured its transcription and translation in yeast. We found that G4s had an inhibitory effect on translation in vivo since "mutants" (where guanines were replaced by adenines in PQS) showed higher expression levels than wild-types. In parallel, we confirmed by circular dichroism measurements that the selected sequences can indeed adopt G4 conformation in vitro. Analysis of RNA-Seq of polyA RNA in maize seedlings grown in the presence of a G4-stabilizing ligand (NMM) showed both inhibitory as well as stimulatory effects on the transcription of LTR retrotransposons. CONCLUSIONS: Our results demonstrate that quadruplex DNA located within long terminal repeats of LTR retrotransposons can be formed in vivo and that it plays a regulatory role in the LTR retrotransposon life-cycle, thus also affecting genome dynamics.

• MeSH
• G-kvadruplexy * MeSH
• genetická transkripce MeSH
• genom rostlinný * MeSH
• koncové repetice * MeSH
• kukuřice setá genetika   růst a vývoj   metabolismus   MeSH
• reportérové geny * MeSH
• retroelementy * MeSH
• Saccharomyces cerevisiae genetika   růst a vývoj   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The prospective study dealing with the recurrence and progression of superficial bladder cancer prediction on the basis of expression profiles of candidate genes PAX5, HSP60, BCL2 and BAX in tumour biopsies. Another aim of the study is to develop methodology of semiquantitative immunohistochemical examination of PAX5 experession, to find out the correlation with the expression detected by RT-PCR and put PAX5 immunohistochemistry into clinical practice. Next aim of the study is to find out if the immunohistochemical examination of p53, Ki-67 and PAX5 expression in non-tumour mucosa samples taken during the superficial bladder carcinoma transurethral resection brings any prognostic information. The study will bring a suggestion of using the prognostic markers when following patients with superficial bladder carcinoma and when planning the treatment, which will be benefical for them.

Prospektivní studie zabývající se predikcí výskytu recidiv a progrese povrchových NMM za základě stanovení expresních profilů kandidátních genů PAX5, HSP60, BCL2 a BAX v tkáni karcinomu močového měchýře. Dalším cílem je rozpracovat metodiku semikvantitativního způsobu hodnocení imunohistochemického vyšetření exprese PAX5, zjistit korelaci s mírou exprese zjišťovanou pomocí RT-PCR a zavést imunohistochemii PAX5 do klinické praxe. Dalším cílem je zjistit zda imunohistochemické vyšetření exprese p53, Ki-67a PAX5 ve vzorcích nenádorové sliznice, odebírané při transuretrální resekci povrchového NMM, přináší prognostickou informaci stran recidivy či progrese onemocnění. Studie přinese návrh použití prognostických markerů při sledování a plánování léčby pacientů s povrchovým nádorem měchýře, což bude mít přínos pro kvalitu pacientova života i ekonomický.

• MeSH
• aktivátorový protein specifický pro B-buňky analýza   MeSH
• chaperon hsp60 analýza   MeSH
• exprese genu imunologie   MeSH
• imunohistochemie metody   využití   MeSH
• nádory močového měchýře MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   využití   MeSH
• prognóza MeSH
• protein X asociovaný s bcl-2 analýza   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• urologie
• onkologie
• biologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Guanine-rich nucleic acid sequences can fold into four-stranded G-quadruplex (G4) structures. Despite growing evidence for their biological significance, considerable work still needs to be done to detail their cellular occurrence and functions. Herein, we describe an optimized core-extended naphthalene diimide (cex-NDI) to be exploited as a G4 light-up sensor. The sensing mechanism relies on the shift of the aggregate-monomer equilibrium towards the bright monomeric state upon G4 binding. In contrast with the majority of other ligands, this novel cex-NDI is able to discriminate among G4s with different topologies, with a remarkable fluorescent response for the parallel ones. We investigate this sensing by means of biophysical methods, comparing the lead compound to a non-selective analogue. We demonstrate that mitigating the affinity of the binding core for G4s results in an increased selectivity and sensitivity of the fluorescent response. This is achieved by replacing positively charged substituents with diethylene glycol (DEG) side chains. Remarkably, the limit of detection values obtained for parallel G4s are more than one order of magnitude lower than those of the parallel-selective ligand N-methyl mesoporphyrin IX (NMM). Interestingly, the classical fluorescent intercalator displacement (FID) assay failed to reveal binding of cex-NDI to G4 because of the presence a ternary complex (G4-TO-cex-NDI) revealed by electrospray-MS. Our study thus provides a rational basis to design or modify existent scaffolds to redirect the binding preference of G4 ligands.

• MeSH
• biosenzitivní techniky metody   MeSH
• fluorescenční barviva chemická syntéza   chemie   farmakologie   MeSH
• G-kvadruplexy * účinky léků   MeSH
• imidy chemická syntéza   chemie   farmakologie   MeSH
• interkalátory chemická syntéza   chemie   farmakologie   MeSH
• ligandy * MeSH
• naftaleny chemická syntéza   chemie   farmakologie   MeSH
• rozpustnost MeSH
• substrátová specifita MeSH
• vazebná místa MeSH
• voda chemie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Aortic dissection is a biomechanical phenomenon associated with a failure of internal cohesion, which manifests itself through the delamination of the aortic wall. The goal of this study is to deepen our knowledge of the delamination strength of the aorta. To achieve this, 661 peeling experiments were carried out with strips of the human aorta collected from 46 cadavers. The samples were ordered into groups with respect to (1) anatomical location, (2) orientation of the sample, and (3) extension rate used within the experiment. The obtained results are in accordance with the hypothesis that delamination resistance is not sensitive to the extension rates 0.1, 1, 10, and 50 mms-1. We arrived at this conclusion for all positions along the aorta investigated in our study. These were the thoracic ascending (AAs), thoracic descending (ADs), and the abdominal aorta (AAb), simultaneously considering both the longitudinal (L) as well as the circumferential (C) orientations of the samples. On the other hand, our results showed that the delamination strength differs significantly with respect to the anatomical position and orientation of the sample. The medians of the delamination strength were as follows, 4.1 in AAs-L, 3.2 in AAs-C, 3.1 in ADs-L, 2.4 in ADs-C, AAb-L in 3.6, and 2.7 in AAb-C case (all values are in 0.01·Nmm-1). This suggests that resistance to crack propagation should be an anisotropic property and that the aorta is inhomogeneous along its length from the point of view of delamination resistance. Finally, correlation analysis proved that the delamination strength of the human aorta significantly decreases with age.

• MeSH
• anizotropie MeSH
• aorta abdominalis MeSH
• aorta thoracica MeSH
• biomechanika MeSH
• disekce aorty * MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cieľ: Na dosiahnutie najlepších výsledkov liečby u pacientov s vysoko rizikovými svalovinu neinfiltrujúcimi nádormi močového mechúra (NMM) je nevyhnutná udržiavacia intravezikálna BCG liečba trvajúca najmenej 1 rok. S rastúcim počtom instilácií stúpa aj výskyt nežiaducich účinkov. Uvádzame naše skúsenosti s dlhodobou BCG liečbou, jej tolerabilitou, výskytom komplikácií a ich liečbou. Materiál a metodika: V rokoch 1997 až 2007 sme indikovali na intravezikálnu BCG liečbu (kmeň Connaught) 132 pacientov s vysoko rizikovými Ta-T1-CIS NMM. Indikáciou na BCG liečbu boli: G3 nádory, CIS, multilokulárne T1G1 - 2, ≥ 2 recidívy počas jedného roka sledovania. Liečba po 6-týždňovej indukčnej fáze pokračovala minimálne 1-ročnou udržiavacou liečbou s podávaním BCG raz týždenne počas 3 týždňov za 3, 6, 12 mesiacov a potom každých 6 mesiacov od zahájenia liečby. V tejto retrospektívnej štúdii sme vyhodnotili zdravotné záznamy 83 pacientov, ktorí absolvovali 1-ročnú udržiavaciu liečbu, alebo ju prerušili kvôli jej intolerancii alebo závažným komplikáciám. Pacienti, ktorí prerušili liečbu z iných dôvodov (napr. zlyhanie liečby), neboli do štúdie zaradení. Tolerabilitu liečby sme hodnotili z klinického pohľadu, ako výskyt a závažnosť vedľajších účinkov, nutnosť odkladu instilácií, zníženia dávky BCG (na 27 mg) alebo definitívneho prerušenia liečby. Výsledky: Z 83 pacientov absolvovalo 1-ročnú intravezikálnu liečbu presne podľa schémy (6 + 3 + 3 + 3 instilácie) a s prijateľnou tolerabilitou 52 (63 %) pacientov (nežiaduce účinky grade 0 alebo 1 trvajúce menej ako 48 hodín). Ďalší 20 pacienti (24 %) absolvovali celú schému 1-ročnej liečby s nutnosťou odkladov instilácií (9 p.), zníženia dávky na 27 mg (5 p.) alebo ich kombináciou (6 p.) kvôli nežiaducim účinkom grade 2 trvajúcim viac ako 48 hodín. Šiesti pacienti (7 %) odmietli pokračovať v liečbe kvôli jej intolerancii (urgencia, cystalgie, recidivujúce infekcie), ale bez závažných nežiaducich účinkov. Len 5 pacientov (6 %) prerušilo iniciálnu alebo udržiavaciu liečbu kvôli závažným nežiaducim účinkom grade 3 alebo 4 (1 prípad BCG hepatitídy a pneumonitídy, 1 prípad BK+ cystitídy, 1 prípad granulomatóznej epididymitídy a perianálneho abscesu, 1 prípad reaktívnej artritídy a 1 prípad granulomatóznych lézií na penise). Všetky nežiaduce účinky boli úspešne liečené podľa publikovaných odporúčaní bez trvalých následkov. Závery: V našej štúdii 72 pacientov (86 %) absolvovalo minimálne 1-ročnú udržiavaciu intravezikálnu BCG liečbu s dobrou tolerabilitou, 6 pacientov v nej odmietlo pokračovať kvôli zlej tolerabilite a len u 5 pacientov musela byť liečba prerušená kvôli závažným komplikáciám. Intravezikálna BCG liečba je bezpečná, dobre tolerovaná, s prijateľnou kvalitou života a primeraným pomerom benefitu a rizika.

Objective: The maintenance intravesical BCG therapy continuing at least 1 year is necessary to achieve the best results in patients with high risk non-muscle invasive bladder cancer. With the number of instillations the incidence of adverse events increases. Our experience with longterm intravesical BCG therapy, its tolerability, the incidence of complications and their treatment is reported. Material and methods: Between 1997 and 2007 132 patients with high risk Ta-T1-CIS bladder cancer were indicated for intravesical BCG therapy (strain Connaught) at our department. There were following indications for BCG treatment: any G3 tumour, CIS, multifocal T1G1 - 2, at least two recurrences within the most recent year. The treatment started with 6-week induction therapy and continued with minimum 1-year maintenance therapy with BCG instillations each week for 3 weeks given 3, 6, 12 months and then every 6 months after initial treatment. In this retrospective study we evaluated medical records from 83 patients who achieved 1-year maintenance therapy or interrupted therapy due to its intolerability or significant complications. Patients who interrupted treatment due to other reasons (e.g. treatment failure) were not included in this study. Treatment tolerability was evaluated from clinical view as incidence and relevance of side effects, necessity to postpone instillations, to reduce BCG dose (27 mg) or to interrupt treatment definitely. Results: 52 from 83 evaluated patients (63 %) treated with intravesical BCG completed at least 1-year lasting regimen (6 + 3 + 3 + 3 instillations) according to schedule with acceptable tolerability (according to WHO scale for side effects grade 0 or 1, lasting < 48 hours). Other 20 patients (24 %) completed 1-year treatment regimen with temporary deferred instillations (9 pts), reduced dose to 27 mg (5 pts) or combination of both (6 pts) (grade 2 side effects lasting > 48 hours). 6 pts (7 %) refused maintenance therapy during first year due to intolerability (urgency, bladder pain, recurrent infection), but without serious side effects. Only 5 pts (6 %) discontinued initial or maintenance therapy due to serious side effects grade 3 or 4 (1 for BCG hepatitis + pneumonitis, 1 for BK+ cystitis, 1 for granulomatous epididymitis and perianal abscess, 1 for reactive arthritis, 1 for granulomatous penis lesions). All side effects were successfully treated according to published recommendations without persistent medical consequences. Conclusions: In our study cohort 72 patients (86 %) were able to complete at least 1-year maintenance intravesical BCG therapy with good tolerability, 6 patients refused maintenance therapy due to poor tolerability and only 5 patients discontinued therapy due to serious complications. Intravesical BCG therapy is safe, well-tolerated treatment modality with acceptable quality of life and reasonable risk/benefit ratio.

OBJECTIVES: Real-world evidence is needed to inform treatment strategies for patients with PsA and axial SpA (axSpA) who have non-musculoskeletal manifestations (NMMs), various risk factors and comorbidities. International collaboration is required to ensure statistical power and to enhance generalizability. The first step forward is identifying which data are currently being collected. Across 17 registries participating in the European Spondyloarthritis Research Collaboration (EuroSpA), we aimed to map recording practices for NMMs, comorbidities and safety outcomes in patients with PsA and axSpA. METHODS: Through a survey with 4,420 questionnaire items, we explored the recording practices of 58 pre-defined conditions (i.e. NMMs, comorbidities and safety outcomes) covering 10 disease areas. In all registries we mapped for each condition whether it was recorded, the recording procedure and the potential to identify it through linkage to other national registries. RESULTS: Conditions were generally recorded at entry into the registry and clinical follow-up visits using a pre-specified list or a coding system. Most registries recorded conditions within the following disease areas: NMMs (number of registries, n = 15-16), cardiovascular diseases (n = 10-14), gastrointestinal diseases (n = 12-13), infections (n = 10-13) and death (n = 14). Nordic countries had the potential for data linkage and generally had limited recording of conditions in their registry, while other countries had comprehensive recording practices. CONCLUSION: A wide range of conditions were consistently recorded across the registries. The recording practices of many conditions and disease areas were comparable across the registries. Our findings support the potential for future collaborative research.

• Publikační typ
• časopisecké články MeSH