• MeSH
• Molecular Diagnostic Techniques MeSH
• Neoplasms * diagnosis   genetics   MeSH
• Gene Expression Profiling MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• molekulární biologie, molekulární medicína
• onkologie
• NML Publication type
• kolektivní monografie

• MeSH
• Genome-Wide Association Study MeSH
• Cholesteatoma, Middle Ear MeSH
• Gene Expression MeSH
• Prospective Studies MeSH
• Oligonucleotide Array Sequence Analysis MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• otorinolaryngologie
• genetika, lékařská genetika
• NML Publication type
• studie

Gen nm 23 má podle některých studií antimetastatickou funkci, výsledky zkoumání antimetastatického efektu jsou však podle literatury rozporuplné, a to i u stejných typů maligních nádorů. V sestavě 77 plicních karcinomů jsme neprokázali vztah exprese genu nm 23 k histologickému typu nádoru ani ke stupni diferenciace nádoru. Nezjistili jsme ani souvislost exprese nm 23 s výskytem metastáz v operačních resektátech plicních karcinomů. Ve skupině adenokarcinomů jsme však pozorovali určitou negativní vazbu exprese nm 23 na vývoj pozdějších metastáz a na snizene přežívaní pacientu. U dlaždicobuněčných, neuroendokrinních a nediferencovaných karcinomů nebyla tato vazba pozorována. Nečekaně nízké pozitivity nebo i negativity nm 23 u málo pokročilých nádorů a sporadické vysoké pozitivity u některých plicních karcinomů s metastázami mohou být vysvětleny mutacemi genu nm 23 nebo jeho inaktivací. Výsledky studie neumožňují využít nm 23 jako spolehlivý prognostický marker u plicních karcinomů.

Gene nm 23 may have an antimetastatic function but relevant data in the literature are ambiguous even in the same types of malignant tumors. In a group of 77 lung carcinomas there was not found any relation of gene nm 23 expression to the histologic type of tumor or the level of its differentiation and to the finding of metastases in resected lung materials either. Adenocarcinomas showed a negative relation of nm 23 expression to later occurence of metastases and to a decreased survival of patients which squamous, neuroendocrine and undifferentiated carcinomas did not. Unexpected low expression or negativity of nm 23 in tumors of little progression and mutations or inactivation. Results of the study do not encourage the usage of nm 23 as a reliable prognostic marker in lung carcinomas.

• MeSH
• Gene Expression MeSH
• Carcinoma MeSH
• Humans MeSH
• Neoplasm Metastasis diagnosis   MeSH
• Lung Neoplasms diagnosis   MeSH
• Oncogene Proteins MeSH
• Check Tag
• Humans MeSH

Východisko. Leukémie vznikají v důsledku defektů v regulaci proliferace, diferenciace a apoptózy, ke kterým dochází v kmenových buňkách nebo progenitorech hematopoézy. Tyto procesy mají několik styčných bodů a jedním z nich jsou inhibitory cyklin-dependentních kináz. Předmětem studie bylo sledování exprese dvou významných regulátorů buněčného cyklu – univerzálních inhibitorů cyklin-dependentních kináz p21 Cip a p27 Kip a exprese proliferačního antigenu Ki-67 v lidských leukemických buňkách a leukocytech zdravých jedinců. Metody a výsledky. Exprese inhibitorů cyklin-dependentních kináz byla sledována na úrovni mRNA především metodou komparativní reverzně-transkriptázové a u vybraných vzorků i metodou kvantitativní řetězové polymerázové reakce v reálném čase. Zatímco exprese p27 Kip byla v leukocytech leukemických pacientů i zdravých jedinců v podstatě univerzální, zjistili jsme velké rozdíly v expresi p21 Cip jak mezi jednotlivými pacienty téhož typu leukémie, tak i mezi jednotlivými typy leukémií a zdravými jedinci. Exprese p21 Cip byla signifikantně vyšší u akutních leukémií (myeloidních i lymfoblastických) než u chronických leukémií (myeloidní i lymfoidní) a zdravých jedinců. Srovnání míry exprese p21 Cip s klinickými parametry pacientů ukázalo, že skupina 14 pacientů akutních myeloidních leukémií žijících déle než 30 měsíců měla signifikantně nižší expresi p21 Cip než skupina 12 pacientů s tímto typem leukémie, kteří nepřežili tento časový interval. Výsledky získané na menšímsouboru pacientů s akutní promyelocytární leukémií (akutní myeloidní leukémie M3) naznačily, že nižší exprese p21 Cip je spojena s příznivějším klinickým průběhem (snazším navozením remise apod.). Závěry. Získané výsledky ukazují na význam inhibitoru cyklin-dependentních kináz p21 Cip u lidských leukémií a na to, že nižší exprese p21 Cip by mohla být pozitivním prognostickým faktorem u pacientů s akutní myeloidní leukémií.

Background. Leukemias develop due to defects in proliferation, differentiation and apoptosis, which take place in stem cells or progenitors of hematopoiesis. These processes have several crossing points, one of them is the role of inhibitors of cyclin-dependent kinases. The aim of this study was to study the expression of cyclin-dependent kinases inhibitors p21 Cip and p27 Kip and expression of proliferative antigen Ki-67 in leukocytes of human leukemia. Methods and Results. The expression of cyclin-dependent kinases inhibitors was detected at mRNA level mainly by comparative reverse-transcription polymerase chain reaction and in selected samples also by the real-time polymerase chain reaction. While p27 Kip expression in leukocytes of leukemic patients and healthy persons was universal, large differences in expression of p21 Cip were found both among individual patients of the same type of leukemia and between different types of leukemias and healthy persons. The p21 Cip expression was significantly higher in acute leukemias than in chronic ones and healthy persons. A comparison of p21 Cip expression with the clinical outcome of the leukemic patients showed that the group of 14 acute leukemia patients surviving more than 30 months had a significantly lower expression of p21 Cip than 12 patients of this type of leukemia who died within this time limit. Moreover, the results obtained on a smaller set of acute promyelocytic leukemia patients indicated that the lower p21 expression is connected with a better prognosis. Conclusion. Our results pointed out the importance of the cyclin-dependent kinase inhibitor p21 Cip in human leukemias and indicated that the lower p21 Cip expression might be a positive prognostic factor in acute myeloid leukemia patients.

• MeSH
• Cell Division MeSH
• Cyclin-Dependent Kinases antagonists & inhibitors   genetics   MeSH
• Gene Expression MeSH
• Research Support as Topic MeSH
• Leukemia diagnosis   genetics   physiopathology   MeSH
• Leukocytes MeSH
• Humans MeSH
• Reverse Transcriptase Polymerase Chain Reaction MeSH
• Proliferating Cell Nuclear Antigen genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Review MeSH
• Comparative Study MeSH

• MeSH
• Antigens, Surface analysis   MeSH
• Fluorescent Antibody Technique methods   MeSH
• Flow Cytometry methods   MeSH
• Blood Platelets MeSH

Cíl práce: Zhodnotit přítomnost exprese genů PAX5 a Shb ve tkáni povrchových uroteliálních karcinomů močového měchýře, posoudit její korelaci s patologií genu p53 a uvážit prognostický význam jednotlivých faktorů. Pacienti a metodika: Do studie bylo zahrnuto 61 pacientů s histologicky prokázaným povrchovým uroteliálním karcinomem močového měchýře. Exprese mRNA genů PAX5 a Shb ve tkáni nádoru byla detekována pomocí reverzní polymerázové řetězové reakce (RT-PCR) a výsledek byl vyjádřen semikvantitativně. Přítomnost mutací p53 byla zachycena pomocí SSCP (single strand conformation polymorphism) a potvrzena sekvenováním. K imunohistochemickému vyšetření p53 byla použita protilátka DO1, k vyjádření výsledku bylo použito semikvantitativní hodnocení pomocí HSCORE (HS). Kontrolní skupinu pro posouzení PAX5 a Shb exprese tvořilo 8 mužů, u kterých byl vzorek urotelu odebrán během operace pro benigní hyperplazií prostaty. Výsledky: Přítomnost exprese PAX5 a Shb byla prokázána u 50, respektive 52 pacientů s uroteliálním nádorem, ale u žádného z kontrolní skupiny. Mutace p53 byla zachycena u jediného pacienta s tumorem pTaG2 a byla lokalizována v exonu 5 (delece prolinu 128). Jaderná imunoreaktivita p53 byla přítomna u většiny pacientů, při použití prahové hodnoty HS 200 mělo 56,9 % nemocných pozitivní nález. Kvantita imunohistochemické pozitivity p53 nekorelovala s kvantitou exprese PAX5. Při použití prahových hodnot HS 200 pro p53 a 0,2 pro PAX5 mělo z 8 progredujících pacientů 7 nadprahovou hodnotu HS a 4 nadprahovou hodnotu PAX5. Závěr: Exprese genu PAX5 je častým nálezem u povrchových uroteliálních karcinomů močového měchýře.

The objective of the work: To evaluate the presence of the PAX5 and Shb genes expression in the tissue of superficial urotelial urinary bladder cancers, to judge its correlation with the p53 gene pathology and to consider the prognostic value of individual factors. Patients and Method: 61 patients with histologically proven superficial urotelial bladder cancer were included into the study. The mRNA expression of PAX5 and Shb genes in the cancer tissue was detected by the reverse polymerase chain reaction (RT- PCR) and the result was expressed semiquantitatively. The presence of p53 mutations was recorded by SSCP (single strand conformation polymorphism) and confirmed by sequening. The immunohistological examination of p53 was made by using DO1 antibody and the result was expressed using semiquantitative evaluation by HSCORE (HS). 8 men created the control group for PAX5 and Shb expression evaluation. Their urotel sample was withdrawn during the operation of benign prostate hyperplasia. Results: The presence of PAX5 and Shb expression was proven in 50, let us say in 52 patients with urotelial cancer, but it was not proven in anybody of the control group. p53 mutation was recorded in one patient with pTaG2 tumour and it was localised in 5 exon (proline 128 deletion). Nuclear immunoreactivity of p53 was present in the majority of patients, 56,9% of patients had positive finding using the threshold value HS 200. The quantity of p53 immunohistochemical positivity did not correlate with the quantity of PAX5 expression. Using the threshold values of HS 200 for p53 and 0,2 for PAX5 the 7 of 8 progressing patients had supra-threshold HS value and 4 of them had supra-threshold PAX5 value. Conclusion: The PAX5 gene expression is an often finding in superficial urotelial urinary bladder cancers.

• MeSH
• Research Support as Topic MeSH
• Genes, p53 genetics   MeSH
• Humans MeSH
• Mutation genetics   MeSH
• Urinary Bladder Neoplasms diagnosis   genetics   MeSH
• Gene Expression Profiling methods   utilization   MeSH
• Check Tag
• Humans MeSH

Twelve human embryos and fetuses aged of 7-30 weeks of intrauterinal life were examined to determine the expression of bcl-2 gene in the developing kidney. Tissue samples were routinely processed and three-step indirect immunohistochemical method was used for the detection of Bcl-2 protein. End-point cytophotometry was performed with computer-controlled microscope photometer with a scanning table and the mean relative absorbance of the final product of peroxidase reaction was determined and taken as a measure of Bcl-2 expression. The morphometric evaluation was carried out from the TV display using Weibel s universal hexagonal raster and we determined the relative volume of Bcl-2 positive structures in the various zones of the embryonal kidney. The aim of our research was mapping of the Bcl-2 occurrence in the developing kidney of human embryos and fetuses. The Bcl-2 protein is involved in the regulation of apoptosis and its effect is antiapoptotic. The highest Bcl-2 expression was proved in the cells of metanephrogenic blastema. The lower occurrence of Bcl-2 positive cells was demonstrated in proximal tubules analges+ and it was almost on the borderline of detection in branches of ureteral bud. In the fetal period the marked Bcl-2 expression was maintained in the epithelial cells of proximal tubules analges.

• MeSH
• Cytophotometry MeSH
• Kidney Medulla embryology   MeSH
• Embryo, Mammalian MeSH
• Embryonic and Fetal Development * MeSH
• Genes, bcl-2 * MeSH
• Gestational Age MeSH
• Kidney Cortex embryology   MeSH
• Kidney cytology   embryology   MeSH
• Humans MeSH
• Fetus MeSH
• Proto-Oncogene Proteins c-bcl-2 analysis   MeSH
• Gene Expression Regulation, Developmental * MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Antigens, Neoplasm MeSH
• Cell Line MeSH
• Cytological Techniques MeSH
• Fluorescent Antibody Technique MeSH
• Hematopoietic Stem Cells immunology   MeSH
• Isoantigens MeSH
• Lymphoma immunology   MeSH

The use of RT-qPCR provides a powerful tool for gene expression studies; however, the proper interpretation of the obtained data is crucially dependent on accurate normalization based on stable reference genes. Recently, strong evidence has been shown indicating that the expression of many commonly used reference genes may vary significantly due to diverse experimental conditions. The isolation of pancreatic islets is a complicated procedure which creates severe mechanical and metabolic stress leading possibly to cellular damage and alteration of gene expression. Despite of this, freshly isolated islets frequently serve as a control in various gene expression and intervention studies. The aim of our study was to determine expression of 16 candidate reference genes and one gene of interest (F3) in isolated rat pancreatic islets during short-term cultivation in order to find a suitable endogenous control for gene expression studies. We compared the expression stability of the most commonly used reference genes and evaluated the reliability of relative and absolute quantification using RT-qPCR during 0-120 hrs after isolation. In freshly isolated islets, the expression of all tested genes was markedly depressed and it increased several times throughout the first 48 hrs of cultivation. We observed significant variability among samples at 0 and 24 hrs but substantial stabilization from 48 hrs onwards. During the first 48 hrs, relative quantification failed to reflect the real changes in respective mRNA concentrations while in the interval 48-120 hrs, the relative expression generally paralleled the results determined by absolute quantification. Thus, our data call into question the suitability of relative quantification for gene expression analysis in pancreatic islets during the first 48 hrs of cultivation, as the results may be significantly affected by unstable expression of reference genes. However, this method could provide reliable information from 48 hrs onwards.

• MeSH
• Insulin secretion   MeSH
• Rats MeSH
• Islets of Langerhans metabolism   secretion   MeSH
• Gene Expression Profiling * MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

1. Nucleotide analogues comprise an important class of drugs used in treatment of viral infections but also cancer. These drugs affect the structural integrity of DNA and activate different pathways and processes in the cell and may directly or indirectly influence the drug metabolizing system. Adefovir dipivoxil (AD) and tenofovir disoproxil (TD) are nucleotide analogues approved for the treatment of chronic hepatitis B and/or HIV/AIDS infection. 2. To evaluate the risk of their drug-drug interactions on the level of drug metabolism, an effect of both compounds on cytochromes P450 expression was studied using cDNA microarrays, real-time RT-PCR and immunoblotting. Mice were given intraperitoneally 25 mg/kg of AD or TD, respectively. As a positive control, a combination of prototypic cytochromes P450 (CYP) inducers, phenobarbital and β-naphthoflavone was chosen. 3. The data obtained showed a significant CYP induction in the positive control group, but no clinically significant induction of CYP genes by AD or TD was observed. Our results support the evidence of safety of AD and TD with respect to drug-drug interactions based on enzyme induction. These findings are important as a plethora of new antivirals of different types are being tested and introduced to clinical practice, mostly to be used in combinations.

• MeSH
• Adenine analogs & derivatives   metabolism   MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Mice, Inbred C57BL MeSH
• Organophosphonates chemistry   metabolism   MeSH
• Gene Expression Regulation, Enzymologic * MeSH
• Oligonucleotide Array Sequence Analysis MeSH
• Gene Expression Profiling MeSH
• Cytochrome P-450 Enzyme System genetics   metabolism   MeSH
• Blotting, Western MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH