The complexity of Neolithic population movements and their interpretation through material culture have been the subject of archaeological research for decades. One of the dominant narratives proposes that groups from the Starčevo-Körös-Criş complex spread from the central towards the northern Balkans in the Early Neolithic and eventually brought the Neolithic lifestyle into present-day Hungary. Broad geographical migrations were considered to shape the continuous expansion of Neolithic groups and individuals. However, recent archaeological research, aDNA, and isotope analyses challenged the synchronous appearance of specific material culture distributions and human movement dynamics through emphasizing communication networks and socio-cultural transformation processes. This paper seeks to retrace the complexity of Neolithic mobility patterns across Hungary by means of strontium and oxygen stable isotope analyses, which were performed on a total of 718 human dental enamel samples from 55 Neolithic sites spanning the period from the Starčevo to the Balaton-Lasinja culture in Transdanubia and from the Körös to the Tiszapolgár cultural groups on the Great Hungarian Plain (Alföld). This study presents the largest strontium and oxygen isotope sample size for the Neolithic Carpathian Basin and discusses human mobility patterns on various geographical scales and throughout archaeological cultures, chronological periods, and sex and gender categories in a multiproxy analysis. Based on our results, we discuss the main stages of the Neolithisation processes and particularly trace individual movement behaviour such as exogamy patterns within extensive social networks. Furthermore, this paper presents an innovative differentiation between mobility patterns on small, micro-regional, and supra-regional scales, which provides new insights into the complex organisation of Neolithic communities.

• MeSH
• časové faktory MeSH
• izotopové značení * MeSH
• izotopy kyslíku analýza   MeSH
• lidé MeSH
• stroncium analýza   MeSH
• velikost vzorku MeSH
• zeměpis MeSH
• zubní sklovina chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Maďarsko MeSH

Stable isotope probing (SIP) approaches are a suitable tool to identify active organisms in bacterial communities, but adding isotopically labeled substrate can alter both the structure and the functionality of the community. Here, we validated and demonstrated a substrate-independent protein-SIP protocol using isotopically labeled water that captures the entire microbial activity of a community. We found that 18O yielded a higher incorporation rate into peptides and thus comprised a higher sensitivity. We then applied the method to an in vitro model of a human distal gut microbial ecosystem grown in two medium formulations, to evaluate changes in microbial activity between a high-fiber and high-protein diet. We showed that only little changes are seen in the community structure but the functionality varied between the diets. In conclusion, our approach can detect species-specific metabolic activity in complex bacterial communities and more specifically to quantify the amount of amino acid synthesis. Heavy water makes possible to analyze the activity of bacterial communities for which adding an isotopically labeled energy and nutrient sources is not easily feasible. SIGNIFICANCE: Heavy stable isotopes allow for the detection of active key players in complex ecosystems where many organisms are thought to be dormant. Opposed to the labelling with energy or nutrient sources, heavy water could be a suitable replacement to trace activity, which has been shown for DNA and RNA. Here we validate, quantify and compare the incorporation of heavy water either labeled with deuterium or 18‑oxygen into proteins of Escherichia coli K12 and of an in vitro model of a human gut microbial ecosystem. The significance of our research is in providing a freely available pipeline to analyze the incorporation of deuterium and 18‑oxygen into proteins together with the validation of the applicability of tracing heavy water as a proxy for activity. Our approach unveils the relative functional contribution of microbiota in complex ecosystems, which will improve our understanding of both animal- and environment-associated microbiomes and in vitro models.

• MeSH
• izotopové značení MeSH
• izotopy uhlíku analýza   MeSH
• lidé MeSH
• mikrobiota * MeSH
• oxid deuteria MeSH
• proteiny * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Herein, we demonstrate an efficient method for multi-deuterium labelling of pirtobrutinib-a Bruton's tyrosine kinase inhibitor recently approved by the FDA-using a straightforward hydrogen isotope exchange (HIE) reaction. A remarkably high level of deuterium incorporation was achieved using an excess of a Kerr-type iridium catalyst. The key factor in the significant deuterium labelling was the decision to employ a deuterium uniformly labelled solvent, chlorobenzene-d5, at an elevated temperature. Virtually, no d0-d3 species were detected, with only traces of d4-d5 isotopomers (< 5%) observable in the mass spectrum of pirtobrutinib-d8, fulfilling requirements for stable isotope-labelled internal standard. The labelled compound-mainly consisting of isotopomers d6-d9 at 82.4% of the total abundance-was isolated in a high yield (73%) and purity (99%). Noteworthy, fluorine group acting as a directing group was observed for the first time. Significant incorporation of deuterium in ortho-positions, exceeding 87%, was observed. Interestingly, chlorinated solvent used in the HIE reactions was non-specifically deuterated yielding up to 0.42 deuterium per chlorobenzene molecule even at an exceptionally low iridium catalyst loading of 4.17 × 10-2 mol%.

• MeSH
• deuterium * chemie   MeSH
• izotopové značení * MeSH
• piperidiny chemie   MeSH
• pyrimidiny chemie   MeSH
• Publikační typ
• časopisecké články MeSH

Deuterium- and tritium-labeled compounds play a principal role in tracing of biologically active molecules in complicated biochemical systems. The state-of-the-art techniques using noble metal catalysts or strong reducing agents often suffers from low functional group tolerances, poor selectivity, tricky or multistep synthesis of reagents, and low specific activity of the labeled product. Herein, we demonstrate a mild and nonmetallic technique of deuteration and tritiation of polarized double bonds, such as carbonyl compounds, yielding labeled alcohols of high specific activities. This one-pot synthesis uses carrier-free hydrogen gas in situ activated by a freshly prepared frustrated Lewis pair, generating reducing reagents. This labeling strategy shows better selectivity and functional group tolerances compared with current reductive methods. Reported is an example of the selective reduction of the aldehyde moiety of 3-acetylbenzaldehyde. What makes this technology groundbreaking is its mildness, selectivity, and generation of limited amount of radioactive waste as almost no byproducts were generated after use of (B(C6 F5 )33 H)(3 HTMP) reducing reagent. Radiochemical purity of desired 3 H-labeled product in a crude reaction mixture was determined of over 94%. This work provides, to the community of radiochemists, a practical protocol for frustrated Lewis pairs (FLP)-assisted deuterium/tritium labeling technology.

• MeSH
• deuterium chemie   MeSH
• izotopové značení metody   MeSH
• katalýza MeSH
• Lewisovy kyseliny chemie   MeSH
• tritium chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Microcystins are cyclic peptide toxins with hepatotoxic and tumor-promoting properties, which are produced in significant quantities (up to tens of μg/L) in freshwater cyanobacterial water blooms. Several studies reported microcystin accumulation in fish with possible food transfer to humans. These compounds are further metabolized to cysteine and glutathione conjugates which can be present in tissues in significant concentrations. In this study, we focused on the development and evaluation of robust and highly sensitive SPE-LC-MS/MS method for the analysis of microcystin conjugates in fish tissue samples. For the first time, we demonstrate the use of isotopically labeled internal standards which are essential for accurate and precise determination of analytes in complex biotic matrices. LLOQs of respective microcystin conjugates (signal-to-noise ratio; S/N > 10, peak-to-peak method) ranged from 3.3 to 5.0 ng/g of tissue fresh weight (FW). The calibration was linear within a range of concentrations from 1 to 70 ng/mL for all analyzed conjugates. The precision and repeatability of the method were very good with recoveries in the range of 88.5-107.6% and relative standard deviations between 8.8 and 13.2% for all analytes. In the follow-up study, fully validated method was used for the determination of microcystin conjugate levels in common carp exposed to microcystin-containing cyanobacterial biomass under controlled conditions. Significant amounts of microcystin conjugates (up to 55 ng/g) were found in the tissues of fish after 7 weeks of exposure. Our method was shown to be robust, sensitive, selective, and suitable for the determination of trace levels of microcystin conjugates in fish tissues.

• MeSH
• biomasa MeSH
• chromatografie kapalinová metody   MeSH
• cystein analýza   MeSH
• glutathion analýza   MeSH
• limita detekce MeSH
• mikrocystiny analýza   chemie   MeSH
• radioisotopová diluční technika MeSH
• reprodukovatelnost výsledků MeSH
• sinice chemie   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH

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

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