In this work aqueous infusions from ten Mentha herbal samples (four different Mentha species and six hybrids of Mentha x piperita) and 20 different peppermint teas were screened by capillary electrophoresis with UV detection. The fingerprint separation was accomplished in a 25 mM borate background electrolyte with 10% methanol at pH 9.3. The total polyphenolic content in the extracts was determined spectrophotometrically at 765 nm by a Folin-Ciocalteu phenol assay. Total antioxidant activity was determined by scavenging of 2,2-diphenyl-1-picrylhydrazyl radical at 515 nm. The peak areas of 12 dominant peaks from CE analysis, present in all samples, and the value of total polyphenolic content and total antioxidant activity obtained by spectrophotometry was combined into a single data matrix and principal component analysis was applied. The obtained principal component analysis model resulted in distinct clusters of Mentha and peppermint tea samples distinguishing the samples according to their potential protective antioxidant effect. Principal component analysis, using a non-targeted approach with no need for compound identification, was found as a new promising tool for the screening of herbal tea products.

• MeSH
• antioxidancia analýza   MeSH
• elektroforéza kapilární MeSH
• léčivé rostliny MeSH
• máta chemie   MeSH
• spektrofotometrie MeSH
• Publikační typ
• časopisecké články MeSH

A high-throughput, medium-discrimination method for preliminary typing and selecting non-identical isolates of lactic acid bacteria in cheeses was developed. RAMP, a PCR with one microsatellite-targeted and one random primer, was used for preliminary typing of 1119 isolates of lactic acid bacteria from Slovak Bryndza cheese. A total of 59 genotypes were identified based on RAMP profiles consisting of 12-23 DNA fragments of 150-3000 bp. For example, 18, 17, 13 and 7 different RAMP-types were identified in Lactobacillus brevis, L. plantarum, L. paracasei and L. fermentum, respectively. The method facilitated well reproducible, medium-discrimination typing of Lactobacillus spp. and Pediococcus spp. at a subspecies level and proved to be suitable for preliminary typing of lactic acid bacteria isolated from cheese.

• MeSH
• acidóza laktátová metabolismus   MeSH
• DNA bakterií genetika   MeSH
• DNA fingerprinting metody   MeSH
• DNA primery genetika   MeSH
• genotyp MeSH
• Lactobacillaceae genetika   izolace a purifikace   klasifikace   MeSH
• mikrosatelitní repetice MeSH
• sýr mikrobiologie   MeSH
• technika náhodné amplifikace polymorfní DNA metody   MeSH

The capability of Fluorescent Random Amplified Microsatellites (F-RAMS) to profile hallucinogenic mushrooms to species and sub-species level was assessed. Fifteen samples of Amanita rubescens and 22 samples of other hallucinogenic and non-hallucinogenic mushrooms of the genera Amanita and Psilocybe were profiled using two fluorescently-labeled, 5'degenerate primers, 5'-6FAM-SpC3-DD (CCA)5 and 5'-6FAM-SpC3-DHB (CGA)5, which target different microsatellite repeat regions. Among the two primers, 5'-6FAM-SpC3-DHB (CGA)5 provided more reliable data for identification purposes, by grouping samples of the same species and clustering closely related species together in a dendrogram based on amplicon similarities. A high degree of intra-specific variation between the 15 A. rubescens samples was shown with both primers and the amplicons generated for all A. rubescens samples were organized into three classes of amplicons (discriminant, private, and marker) based on their individualizing potential.

• MeSH
• Amanita genetika   MeSH
• DNA fingerprinting metody   MeSH
• DNA primery * MeSH
• fluorescence MeSH
• fluorescenční barviva * MeSH
• mikrosatelitní repetice * MeSH
• polymerázová řetězová reakce MeSH
• Psilocybe genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Currently, the interest in microalgae as a source of biologically active components exploitable as supplementary ingredients to food/feed or in cosmetics continues to increase. Existing research mainly aims to focus on revealing and recovering the rare, cost competitive components of the algae metabolom. Because these components could be of very different physicochemical character, a universal approach for their isolation and characterization should be developed. This study demonstrates the systematic development of the extraction strategy that represents one of the key challenges in effective algae bioprospecting, which predefines their further industrial application. By using of Trachydiscus minutus as a model microalgae biomass, following procedures were tested and critically evaluated in order to develop the generic procedure for microalgae bioprospecting: (i) various ways of mechanical disintegration of algae cells enabling maximum extraction efficiency, (ii) the use of a wide range of extraction solvents/solvent mixtures suitable for optimal extraction yields of polar, medium-polar, and non-polar compounds, (iii) the use of consecutive extractions as a fractionation approach. Within the study, targeted screening of selected compounds representing broad range of polarities was realized by ultra-high performance liquid chromatography coupled with high resolution tandem mass spectrometric detection (UHPLC-HRMS/MS), to assess the effectiveness of undertaken isolation steps. As a result, simple and high-throughput extraction-fractionation strategy based on consecutive extraction with water-aqueous methanol-hexane/isopropanol was developed. Moreover, to demonstrate the potential of the UHPLC-HRMS/MS for the retrospective non-target screening and compounds identification, the collected mass spectra have been evaluated to characterize the pattern of extracted metabolites. Attention was focused on medium-/non-polar extracts and characterization of lipid species present in the T. minutus algae. Such detailed information on the composition of native (non-hydrolyzed) lipids of this microalga has not been published yet.

• MeSH
• lipidy analýza   chemie   MeSH
• metabolom MeSH
• metabolomika metody   MeSH
• mikrořasy chemie   metabolismus   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The purpose of this study was to apply a recently introduced proteomic based approach to identify candidate biomarkers of the response to anticancer activity of cyclin-dependent kinase inhibitor, bohemine. Mapping of the total protein expression of CEM lymphoblastic leukemia cells following bohemine treatment was performed by 2-D liquid phase separation. Proteins were fractionated by isoelectric points in pH gradient in the first dimension and each of these pI protein fractions was further separated by hydrophobicity using non-porous silica reverse phase chromatography in the second dimension. 2-D protein expression maps of control untreated and bohemine treated cells were generated and inter-sample comparison was performed. Most of the differentially expressed proteins were present at a decreased level after bohemine treatment while there were four proteins, which were up regulated. These proteins representing candidate biomarkers of cancer cell response to the treatment were selected for identification by mass spectrometry. Our results demonstrating down regulation of three histone variants, different in their pI and hydrophobicity, in response to bohemine indicated that anti-mitotic and anti-cancer activities of this compound may be associated with epigenetic regulation at the level of chromatin structure. Furthermore, crk-like adaptor scaffolding protein represents a new important protein family affected by bohemine. This strategy is valuable for comprehensive proteomic analysis of cellular protein targets and pathways that are relevant to anticancer activity of cyclin-dependent kinase inhibition.

• MeSH
• biologické markery chemie   MeSH
• chromatin chemie   MeSH
• cyklin-dependentní kinasy antagonisté a inhibitory   MeSH
• financování vládou MeSH
• fokální dermální hypoplazie patofyziologie   MeSH
• nádorové biomarkery genetika   MeSH
• nádory metabolismus   MeSH
• protinádorové látky farmakologie   MeSH
• spektrální analýza metody   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

BACKGROUND: Breast carcinomas represent a heterogeneous group of tumors diverse in behavior, outcome, and response to therapy. Identification of proteins resembling the tumor biology can improve the diagnosis, prediction, treatment selection, and targeting of therapy. Since the beginning of the post-genomic era, the focus of molecular biology gradually moved from genomes to proteins and proteomes and to their functionality. Proteomics can potentially capture dynamic changes in protein expression integrating both genetic and epigenetic influences. METHODS: We prepared primary cultures of epithelial cells from 23 breast cancer tissue samples and performed comparative proteomic analysis. Seven patients developed distant metastases within three-year follow-up. These samples were included into a metastase-positive group, the others formed a metastase-negative group. Two-dimensional electrophoretical (2-DE) gels in pH range 4-7 were prepared. Spot densities in 2-DE protein maps were subjected to statistical analyses (R/maanova package) and data-mining analysis (GUHA). For identification of proteins in selected spots, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed. RESULTS: Three protein spots were significantly altered between the metastatic and non-metastatic groups. The correlations were proven at the 0.05 significance level. Nucleophosmin was increased in the group with metastases. The levels of 2,3-trans-enoyl-CoA isomerase and glutathione peroxidase 1 were decreased. CONCLUSION: We have performed an extensive proteomic study of mammary epithelial cells from breast cancer patients. We have found differentially expressed proteins between the samples from metastase-positive and metastase-negative patient groups.

• MeSH
• 2D gelová elektroforéza MeSH
• dospělí MeSH
• epitelové buňky metabolismus   MeSH
• financování organizované MeSH
• lidé středního věku MeSH
• lidé MeSH
• metastázy nádorů patofyziologie   patologie   MeSH
• nádorové buňky kultivované MeSH
• nádorové proteiny metabolismus   MeSH
• nádory prsu metabolismus   MeSH
• peptidové mapování MeSH
• proteomika MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• srovnávací studie MeSH

• MeSH
• genom lidský MeSH
• peptidy MeSH
• proteiny analýza   MeSH
• proteom MeSH
• Publikační typ
• laboratorní příručky MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• genetika, lékařská genetika
• chemie, klinická chemie
• biologie

The translation of metallothioneins (MTs) is one of the defense strategies by which organisms protect themselves from metal-induced toxicity. MTs belong to a family of proteins comprising MT-1, MT-2, MT-3, and MT-4 classes, with multiple isoforms within each class. The main aim of this study was to determine the behavior of MT in dependence on various externally modelled environments, using electrochemistry. In our study, the mass distribution of MTs was characterized using MALDI-TOF. After that, adsorptive transfer stripping technique with differential pulse voltammetry was selected for optimization of electrochemical detection of MTs with regard to accumulation time and pH effects. Our results show that utilization of 0.5 M NaCl, pH 6.4, as the supporting electrolyte provides a highly complicated fingerprint, showing a number of non-resolved voltammograms. Hence, we further resolved the voltammograms exhibiting the broad and overlapping signals using curve fitting. The separated signals were assigned to the electrochemical responses of several MT complexes with zinc(II), cadmium(II), and copper(II), respectively. Our results show that electrochemistry could serve as a great tool for metalloproteomic applications to determine the ratio of metal ion bonds within the target protein structure, however, it provides highly complicated signals, which require further resolution using a proper statistical method, such as curve fitting.

• MeSH
• chlorid sodný chemie   MeSH
• elektrochemie MeSH
• elektrolyty MeSH
• komplexní sloučeniny chemie   metabolismus   MeSH
• kovy chemie   metabolismus   MeSH
• metalothionein chemie   metabolismus   MeSH
• protein - isoformy MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• vazba proteinů MeSH
• Publikační typ
• časopisecké články MeSH

Detection of incipient Alzheimer disease (AD) pathophysiology is critical to identify preclinical individuals and target potentially disease-modifying therapies towards them. Current neuroimaging and biomarker research is strongly focused in this direction, with the aim of establishing AD fingerprints to identify individuals at high risk of developing this disease. By contrast, cognitive fingerprints for incipient AD are virtually non-existent as diagnostics and outcomes measures are still focused on episodic memory deficits as the gold standard for AD, despite their low sensitivity and specificity for identifying at-risk individuals. This Review highlights a novel feature of cognitive evaluation for incipient AD by focusing on spatial navigation and orientation deficits, which are increasingly shown to be present in at-risk individuals. Importantly, the navigation system in the brain overlaps substantially with the regions affected by AD in both animal models and humans. Notably, spatial navigation has fewer verbal, cultural and educational biases than current cognitive tests and could enable a more uniform, global approach towards cognitive fingerprints of AD and better cognitive treatment outcome measures in future multicentre trials. The current Review appraises the available evidence for spatial navigation and/or orientation deficits in preclinical, prodromal and confirmed AD and identifies research gaps and future research priorities.

• MeSH
• Alzheimerova nemoc diagnóza   patofyziologie   MeSH
• biologické markery * MeSH
• lidé MeSH
• prodromální symptomy * MeSH
• prostorová navigace fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

• MeSH
• biologie buňky MeSH
• molekulární biologie MeSH
• Publikační typ
• monografie MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• biologie
• cytologie, klinická cytologie