Ellagic acid, a natural substance found in various fruits and nuts, was previously shown to exhibit beneficial effects towards metabolic syndrome. In this study, using a genetic rat model of metabolic syndrome, we aimed to further specify metabolic and transcriptomic responses to ellagic acid treatment. Adult male rats of the SHR-Zbtb16Lx/k.o. strain were fed a high-fat diet accompanied by daily intragastric gavage of ellagic acid (50 mg/kg body weight; high-fat diet-ellagic acid (HFD-EA) rats) or vehicle only (high-fat diet-control (HFD-CTL) rats). Morphometric and metabolic parameters, along with transcriptomic profile of liver and brown and epididymal adipose tissues, were assessed. HFD-EA rats showed higher relative weight of brown adipose tissue (BAT) and decreased weight of epididymal adipose tissue, although no change in total body weight was observed. Glucose area under the curve, serum insulin, and cholesterol levels, as well as the level of oxidative stress, were significantly lower in HFD-EA rats. The most differentially expressed transcripts reflecting the shift induced by ellagic acid were detected in BAT, showing downregulation of BAT activation markers Dio2 and Nr4a1 and upregulation of insulin-sensitizing gene Pla2g2a. Ellagic acid may provide a useful nutritional supplement to ameliorate features of metabolic syndrome, possibly by suppressing oxidative stress and its effects on brown adipose tissue.

• Klíčová slova
• brown adipose tissue, ellagic acid, insulin resistance, metabolic syndrome, oxidative stress,
• MeSH
• biologické markery analýza   MeSH
• dieta s vysokým obsahem tuků MeSH
• epididymis MeSH
• hnědá tuková tkáň chemie   MeSH
• játra chemie   MeSH
• krevní glukóza analýza   MeSH
• krysa rodu Rattus MeSH
• kyselina ellagová aplikace a dávkování   MeSH
• messenger RNA analýza   MeSH
• metabolický syndrom genetika   metabolismus   prevence a kontrola   MeSH
• oxidační stres účinky léků   MeSH
• potkani inbrední SHR MeSH
• RNA analýza   MeSH
• transkriptom účinky léků   MeSH
• tuková tkáň chemie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• krevní glukóza MeSH
• kyselina ellagová MeSH
• messenger RNA MeSH
• RNA MeSH

Saliva, as a non-invasive and easily accessible biofluid, has been shown to contain RNA biomarkers for prediction and diagnosis of several diseases. However, systematic analysis done by our group identified two problematic issues not coherently described before: (1) most of the isolated RNA originates from the oral microbiome and (2) the amount of isolated human RNA is comparatively low. The degree of bacterial contamination showed ratios up to 1:900,000, so that only about one out of 900,000 RNA copies was of human origin, but the RNA quality (average RIN 6.7 + /- 0.8) allowed for qRT-PCR. Using 12 saliva samples from healthy donors, we modified the methodology to (1) select only human RNA during cDNA synthesis by aiming at the poly(A)+-tail and (2) introduced a pre-amplification of human RNA before qRT-PCR. Further, the manufacturer's criteria for successful pre-amplification (Ct values ≤ 35 for unamplified cDNA) had to be replaced by (3) proofing linear pre-amplification for each gene, thus, increasing the number of evaluable samples up to 70.6%. When considering theses three modifications unbiased gene expression analysis on human salivary RNA can be performed.

• MeSH
• bakteriální RNA genetika   MeSH
• dospělí MeSH
• exprese genu * MeSH
• komplementární DNA genetika   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé MeSH
• RNA ribozomální 18S genetika   MeSH
• RNA analýza   MeSH
• sliny chemie   metabolismus   MeSH
• stanovení celkové genové exprese metody   MeSH
• transkriptom MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriální RNA MeSH
• komplementární DNA MeSH
• RNA ribozomální 18S MeSH
• RNA MeSH

The distinction between lipoma and atypical lipomatous tumor can be challenging in some cases. While detection of MDM2 gene amplification via fluorescence in situ hybridization (FISH) has been well established as a diagnostic tool to distinguish atypical lipomatous tumor and well-differentiated liposarcoma from benign mimics, MDM2 RNA in situ hybridization (RNA-ISH) has recently been proposed as an alternative diagnostic assay. During clinical workup for lipomatous tumors using MDM2 RNA-ISH, we noticed several dysplastic lipomas that were positive for MDM2 RNA-ISH but negative for MDM2 amplification by FISH. In this study, we examined a series of 11 dysplastic lipomas, all confirmed to be negative for MDM2 amplification by FISH. Positive MDM2 RNA-ISH was noted in 10 (91%) dysplastic lipomas. Single-nucleotide polymorphism array on one dysplastic lipoma identified the presence of homozygous deletion of 13q, including the RB1 gene locus with no evidence of MDM2 copy number gain. Our findings on the discordance between MDM2 FISH and MDM2 RNA-ISH highlight the potential utility and pitfalls of using MDM2 RNA-ISH in the distinction of atypical lipomatous tumor and related liposarcomas from dysplastic lipoma.

• Klíčová slova
• Anisometric lipoma, Atypical lipomatous tumor, Dysplastic lipoma, In situ hybridization, MDM2,
• MeSH
• amplifikace genu MeSH
• diferenciální diagnóza MeSH
• dospělí MeSH
• hybridizace in situ metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipom diagnóza   genetika   MeSH
• liposarkom diagnóza   genetika   MeSH
• mladý dospělý MeSH
• nádorové biomarkery analýza   MeSH
• protoonkogenní proteiny c-mdm2 analýza   genetika   MeSH
• RNA analýza   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• MDM2 protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• protoonkogenní proteiny c-mdm2 MeSH
• RNA MeSH

This review summarizes progress in electroanalysis of organic compounds and biomacromolecules by means of bare BDD-based electrodes for the period of 2009-2018. New trends, which have emerged in the reported decade and which have improved their performance in batch voltammetric and amperometric methods and electrochemical detection in liquid flow techniques are commented. Importance of BDD surface termination, effect of boron doping level, and utilization of adsorption of analytes on BDD surfaces enabling development of adsorptive voltammetric techniques are addressed. Further, possibilities of simultaneous determination of analytes by means of voltammetric techniques utilizing computational approaches and multiple-pulse amperometric detection are discussed. Strategies leading to enhancement of sensitivity such as nanostructuring of the BDD surface, fabrication of BDD-based composite materials or new approaches in construction of microelectrodes and microelectrode arrays for biosensing represent another area of interest. Attention is paid to possibilities in detection of amino acids, peptides and proteins, nucleobases, nucleos(t)ides and DNA/RNA.

• Klíčová slova
• Biomolecules, Boron doped diamond, Microelectrodes, Non-enzymatic electroanalysis, Organic compounds, Surface treatment,
• MeSH
• bor chemie   MeSH
• diamant chemie   MeSH
• DNA analýza   MeSH
• elektrochemické techniky přístrojové vybavení   metody   MeSH
• mikroelektrody * MeSH
• organické látky analýza   MeSH
• peptidy analýza   MeSH
• proteiny analýza   MeSH
• RNA analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• bor MeSH
• diamant MeSH
• DNA MeSH
• organické látky MeSH
• peptidy MeSH
• proteiny MeSH
• RNA MeSH

In human cells, the intergenic spacers (IGS), which separate ribosomal genes, are complex approximately 30 kb-long loci. Recent studies indicate that all, or almost all, parts of IGS may be transcribed, and that at least some of them are involved in the regulation of the ribosomal DNA (rDNA) transcription, maintenance of the nucleolar architecture, and response of the cell nucleus to stress. However, since each cell contains hundreds not quite identical copies of IGS, the structure and functions of this locus remain poorly understood, and the dynamics of its products has not been specially studied. In this work, we used quantitative PCR to measure the expression levels of various rDNA regions at different times after inhibition of the transcription by Actinomycin D applied in high doses. This approach allowed us to measure real or extrapolated half-life times of some IGS loci. Our study reveals characteristic dynamic patterns suggestive of various pathways of RNA utilization and decay.

• Klíčová slova
• Intergenic spacer, Processing, RNA decay, lncRNAs, rDNA,
• MeSH
• HeLa buňky MeSH
• lidé MeSH
• mezerníky ribozomální DNA chemie   genetika   metabolismus   MeSH
• RNA analýza   biosyntéza   genetika   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• mezerníky ribozomální DNA MeSH
• RNA MeSH

Osteoblastic differentiation is a multistep process characterized by osteogenic induction of mesenchymal stem cells, which then differentiate into proliferative pre-osteoblasts that produce copious amounts of extracellular matrix, followed by stiffening of the extracellular matrix, and matrix mineralization by hydroxylapatite deposition. Although these processes have been well characterized biologically, a detailed transcriptional analysis of murine primary calvaria osteoblast differentiation based on RNA sequencing (RNA-seq) analyses has not previously been reported. Here, we used RNA-seq to obtain expression values of 29,148 genes at four time points as murine primary calvaria osteoblasts differentiate in vitro until onset of mineralization was clearly detectable by microscopic inspection. Expression of marker genes confirmed osteogenic differentiation. We explored differential expression of 1386 protein-coding genes using unsupervised clustering and GO analyses. 100 differentially expressed lncRNAs were investigated by co-expression with protein-coding genes that are localized within the same topologically associated domain. Additionally, we monitored expression of 237 genes that are silent or active at distinct time points and compared differential exon usage. Our data represent an in-depth profiling of murine primary calvaria osteoblast differentiation by RNA-seq and contribute to our understanding of genetic regulation of this key process in osteoblast biology.

• Klíčová slova
• Alternative splicing, Bone cells, Non-coding RNA, RNAseq, Topological domains,
• MeSH
• alternativní sestřih MeSH
• buněčná diferenciace genetika   MeSH
• kultivované buňky MeSH
• lebka fyziologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• osteoblasty fyziologie   MeSH
• osteogeneze genetika   MeSH
• RNA analýza   MeSH
• stanovení celkové genové exprese MeSH
• transkriptom genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• RNA MeSH

• MeSH
• algoritmy MeSH
• databáze genetické MeSH
• lidé MeSH
• melanom chemie   genetika   mortalita   MeSH
• myši MeSH
• receptory antigenů T-buněk chemie   genetika   MeSH
• receptory antigenů * analýza   genetika   metabolismus   MeSH
• RNA analýza   genetika   MeSH
• sekvenční analýza RNA metody   MeSH
• stanovení celkové genové exprese metody   MeSH
• výpočetní biologie metody   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• receptory antigenů T-buněk MeSH
• receptory antigenů * MeSH
• RNA MeSH

• MeSH
• DNA analýza   MeSH
• falešně pozitivní reakce MeSH
• imunofenotypizace MeSH
• imunologické techniky * MeSH
• lidé MeSH
• proliferace buněk MeSH
• průtoková cytometrie MeSH
• řízení kvality MeSH
• RNA analýza   MeSH
• separace buněk MeSH
• směrnice jako téma * MeSH
• software MeSH
• T-lymfocyty cytologie   MeSH
• výzkumný projekt MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• DNA MeSH
• RNA MeSH

A hallmark of oocyte development in mammals is the dependence on the translation and utilization of stored RNA and proteins rather than the de novo transcription of genes in order to sustain meiotic progression and early embryo development. In the absence of transcription, the completion of meiosis and early embryo development in mammals relies significantly on maternally synthesized RNAs. Post-transcriptional control of gene expression at the translational level has emerged as an important cellular function in normal development. Therefore, the regulation of gene expression in oocytes is controlled almost exclusively at the level of mRNA and protein stabilization and protein synthesis. This current review is focused on the recently emerged findings on RNA distribution related to the temporal and spatial translational control of the meiotic progression of the mammalian oocyte.

• Klíčová slova
• Meiosis, Oocyte, RNA, RNP, Translation,
• MeSH
• lidé MeSH
• meióza MeSH
• oocyty cytologie   metabolismus   MeSH
• oogeneze MeSH
• proteosyntéza * MeSH
• RNA analýza   genetika   MeSH
• transkriptom MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• RNA MeSH

BACKGROUND/AIMS: Subclinical rejection diagnosed from protocol biopsies is thought to be a risk factor of long- term allograft dysfunction. The reason why in some patients subclinical rejection does not represent risk for progression is not fully understood. METHODS: The intragraft expression of 376 target genes involved in chemokine defense, apoptosis, inflammation, tolerance and TGF-β signalling pathways was measured using quantitative real-time RT-PCR (2(-)∆∆(Ct)) method in subclinical inflammation (SCI, n=10), clinical inflammation in acute T-cell mediated rejection (CI, n=10) and no rejection samples (n=9). RESULTS: Clinical inflammation group showed a increased expression of genes for chemotaxis mediating cytokines (CCL1, CCL17, CCL24, CCL25, CCL26), cytokine receptors (CCR1, CCRL2, IL1RAPL2, CXCR5), proinflammatory cytokines (IL12A, LTA), inflammatory mediator (PTAFR), complement protein C3, executioner protein of apoptosis (CASP7), growth factor (TGFA), colony stimulating factor (CSF-2), proteins involved in dendritic cells differentiation and interaction (CD209, LAMP3), regulation of immune response (LILRB2, LILBRB4). The quantitative difference in transcripts signature between SCI and CI is consistent with stronger proinflammatory setting of CI. Prostaglandin E2 receptor gene expression was independently associated with lower risk of further graft function deterioration (OR 0.11, CI 0.01-0.78, p<0.0001). CONCLUSION: Subclinical acute kidney inflammation has transcriptional profile of immune injury of lower extend compared to clinical acute inflammation.

• MeSH
• apoptóza MeSH
• biopsie MeSH
• chemokiny metabolismus   MeSH
• cytokiny metabolismus   MeSH
• dospělí MeSH
• exprese genu genetika   MeSH
• imunosupresiva terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mediátory zánětu metabolismus   MeSH
• přežívání štěpu MeSH
• progrese nemoci MeSH
• receptory cytokinové metabolismus   MeSH
• rejekce štěpu genetika   metabolismus   MeSH
• riziko MeSH
• RNA analýza   biosyntéza   MeSH
• senioři MeSH
• signální transdukce MeSH
• transformující růstový faktor beta metabolismus   MeSH
• transplantace ledvin * MeSH
• výsledek terapie MeSH
• zánět patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chemokiny MeSH
• cytokiny MeSH
• imunosupresiva MeSH
• mediátory zánětu MeSH
• receptory cytokinové MeSH
• RNA MeSH
• transformující růstový faktor beta MeSH