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The current standard method for amino acid signal identification in protein NMR spectra is sequential assignment using triple-resonance experiments. Good software and elaborate heuristics exist, but the process remains laboriously manual. Machine learning does help, but its training databases need millions of samples that cover all relevant physics and every kind of instrumental artifact. In this communication, we offer a solution to this problem. We propose polyadic decompositions to store millions of simulated three-dimensional NMR spectra, on-the-fly generation of artifacts during training, a probabilistic way to incorporate prior and posterior information, and integration with the industry standard CcpNmr software framework. The resulting neural nets take [ 1 H, 13 C] slices of mixed pyruvate-labeled HNCA spectra (different CA signal shapes for different residue types) and return an amino acid probability table. In combination with primary sequence information, backbones of common proteins (GB1, MBP, and INMT) are rapidly assigned from just the HNCA spectrum.

Purpose: The objective of the present work was to test the feasibility of in vivo imaging of hyperpolarized 50-nm silicon-29 ( 29 Si) nanoparticles., Methods: Commercially available, crystalline 50-nm nanoparticles were hyperpolarized using dynamic polarization transfer via the endogenous silicon oxide-silicon defects without the addition of exogenous radicals. Phantom experiments were used to quantify the effect of sample dissolution and various surface coating on T 1 and T 2 relaxation. The in vivo feasibility of detecting hyperpolarized silicon-29 was tested following intraperitoneal, intragastric, or intratumoral injection in mice and compared with the results obtained with previously reported, large, micrometer-size particles. The tissue clearance of SiNPs was quantified in various organs using inductively coupled plasma optical emission spectroscopy., Results: In vivo images obtained after intragastric, intraperitoneal, and intratumoral injection compare favorably between small and large SiNPs. Improved distribution of small SiNPs was observed after intraperitoneal and intragastric injection as compared with micrometer-size SiNPs. Sufficient clearance of nanometer-size SiNPs using ex vivo tissue sample analysis was observed after 14 days following injection, indicating their safe use., Conclusion: In vivo MRI of hyperpolarized small 50-nm SiNPs is feasible with polarization levels and room-temperature relaxation times comparable to large micrometer-size particles., (© 2024 The Author(s). Magnetic Resonance in Medicine published by Wiley Periodicals LLC on behalf of International Society for Magnetic Resonance in Medicine.)

Stable isotopes of carbon, hydrogen, nitrogen, oxygen and sulfur are widespread in nature. Nevertheless, their relative abundance is not the same everywhere. This is due to kinetic isotope effects in enzymes and other physical principles such as equilibrium thermodynamics. Variations in isotope ratios offer unique insights into environmental pollution, trophic relationships in ecology, metabolic disorders and Earth history including climate history. Although classical isotope ratio mass spectrometry (IRMS) techniques still struggle to access intramolecular information like site-specific isotope abundance, electrospray ionization-Orbitrap mass spectrometry can be used to achieve precise and accurate intramolecular quantification of isotopically substituted molecules ('isotopocules'). This protocol describes two procedures. In the first one, we provide a step-by-step beginner's guide for performing multi-elemental, intramolecular and site-specific stable isotope analysis in unlabeled polar solutes by direct infusion. Using a widely available calibration solution, isotopocules of trifluoroacetic acid and immonium ions from the model peptide MRFA are quantified. In the second approach, nitrate is used as a simple model for a flow injection routine that enables access to a diverse range of naturally occurring isotopic signatures in inorganic oxyanions. Each procedure takes 2-3 h to complete and requires expertise only in general mass spectrometry. The workflows use optimized Orbitrap IRMS data-extraction and -processing software and are transferable to various analytes amenable to soft ionization, including metabolites, peptides, drugs and environmental pollutants. Optimized mass spectrometry systems will enable intramolecular isotope research in many areas of biology., (© 2024. Springer Nature Limited.)

Recent advances in the use of stable isotopes necessitate novel synthesis techniques for isotope separation and enrichment that are scalable and offer high throughput. Stable-isotope-enriched nanostructures can offer unique advantages as nanomedicines, safe tracers, and labels and are critical for applications in various industrial processes, metabolic research, and medicine. So far, there exists no method to synthesize miniature isotope-enriched materials at the nanoscale. In this study, an ultrafast Laser-induced isotope enrichment at nanoscale (LIIEN) is put forward to synthesize isotope-enriched nanostructures, eliminating the need for large equipment and expenses, thereby demonstrating a lab-scale isotope enrichment process. A significant isotope enrichment for Carbon nanostructures is observed. The isotope enrichment can be attributed to the redistribution of isotope ions in the plasma plume explained by the plasma centrifuge model. The LIIEN synthesized structures exhibit excellent Surface-Enhanced Raman Scattering (SERS) signal enhancement and reproducibility, making them potential candidates for SERS-based biomolecule sensing. This technique is an efficient method to fabricate nanosized isotope-enriched structures of characteristic properties by carefully tuning laser parameters at ambient conditions., (© 2024 Wiley‐VCH GmbH.)

In earlier quantum chemical calculations of isotope effects, chemical species in the liquid phase were generally treated as existing in the gas phase. In recent years, however, advances in computational programs have made it easier for the self-consistent reaction field (SCRF) method to handle chemical species in the liquid phase, and as a result, it has become easier to apply the SCRF method to isotope effect calculations. This paper concerns the scope of application of the DFT-SCRF method to reversible processes for hydrogen isotope enrichment. It is found that the applicability of the method depends on the type of the intermolecular interaction in the liquid phase and the degree of hydrogen isotope effect (separation factor) on which the process is based. When the magnitude of the isotope effect of the separation system is greater than 10 -1 , the simple SCRF method is fully applicable; when the magnitude is around 10 -2 , SCRF with a dimer model, in which the monomer is replaced by a dimer, is applicable for the analysis of the liquid phase with relatively strong intermolecular interactions. Anharmonic correction to the separation factor calculated based on harmonic frequencies may be effective to systems with the liquid phase with weak intermolecular interactions.

We present a new approach to Cu isotopic measurements using a state-of-the-art Nu Sapphire multicollector inductively coupled plasma source mass spectrometer equipped with a collision/reaction cell (CRC-MC-ICPMS). We investigate the effects of Na doping and Cu concentration mismatch between bracketing standard and unknown samples and demonstrate the efficacy of introducing a He-H 2 gas mix into the CRC to efficiently eliminate the sample matrix-based 40 Ar 23 Na + isobaric interference on 63 Cu + . This capability is crucial when measuring samples with high Na/Cu ratios, such as some biological samples, which have significantly different chemical compositions compared to most geological samples. Moreover, considering the necessity of obtaining large data sets for biological samples to ensure reliable interpretations, the implementation of a CRC for mitigating the 40 Ar 23 Na + interference offers the advantage of minimizing the requirement for extensive Cu chemical separation procedure prior to Cu isotopic measurements. Our results demonstrate that the accurate determination of the δ 65 Cu values is achievable for samples with Na/Cu concentration ratios of up to ∼65, even when measuring 100 ppb Cu solutions (equivalent to a signal of ∼3.5-4 V total Cu). Furthermore, our results showcase a good short-term repeatability on δ 65 Cu for pure Cu standard solutions (NIST SRM 976 and Cu-IPGP), typically of 0.05‰ (2 SD) when measuring >50 ppb Cu solutions. Our long-term external reproducibility stands at approximately 0.07‰ (2 SD). This value accounts for the variable Cu concentrations analyzed across the different analytical sequences (from 10 to 100 ppb Cu solutions). To validate the robustness of our analytical method, we first conduct a comparison between data sets from mice brains processed twice through column chemistry using a Thermo Finnigan Neptune MC-ICPMS and a Nu Sapphire CRC-MC-ICPMS in CRC mode. This comparison serves to verify the reliability of our method for measuring Cu isotopic composition using the CRC on samples with a low Na/Cu ratio after traditional chemical processing. Then, we compare the data sets obtained for biological standards (tuna fish ERM-CE 464 (IRMM) and human serum Seronorm Trace Elements Serum L-1) processed either once, or twice, through column chemistry and demonstrate that the CRC allows accurate Cu isotopic measurements of the samples processed only once and therefore with a higher Na/Cu ratio.

Origin authentication methods are pivotal in counteracting frauds and provide evidence for certification systems. For these reasons, geographical origin authentication methods are used to ensure product origin. This study focused on the origin authentication (i.e. at the producer level) of a typical mountain cheese origin using various approaches, including shotgun metagenomics, volatilome, near infrared spectroscopy, stable isotopes, and elemental analyses. DNA-based analysis revealed that viral communities achieved a higher classification accuracy rate (97.4 ± 2.6 %) than bacterial communities (96.1 ± 4.0 %). Non-starter lactic acid bacteria and phages specific to each origin were identified. Volatile organic compounds exhibited potential clusters according to cheese origin, with a classification accuracy rate of 90.0 ± 11.1 %. Near-infrared spectroscopy showed lower discriminative power for cheese authentication, yielding only a 76.0 ± 31.6 % classification accuracy rate. Model performances were influenced by specific regions of the infrared spectrum, possibly associated with fat content, lipid profile and protein characteristics. Furthermore, we analyzed the elemental composition of mountain Caciotta cheese and identified significant differences in elements related to dairy equipment, macronutrients, and rare earth elements among different origins. The combination of elements and isotopes showed a decrease in authentication performance (97.0 ± 3.1 %) compared to the original element models, which were found to achieve the best classification accuracy rate (99.0 ± 0.01 %). Overall, our findings emphasize the potential of multi-omics techniques in cheese origin authentication and highlight the complexity of factors influencing cheese composition and hence typicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The identification and in situ cultivation of functional yet uncultivable microorganisms are important to confirm inferences regarding their ecological functions. Here, we developed a new method that couples Raman-activated cell sorting (RACS), stable-isotope probing (SIP), and genome-directed cultivation (GDC)─namely, RACS-SIP-GDC─to identify, sort, and cultivate the active toluene degraders from a complex microbial community in petroleum-contaminated soil. Using SIP, we successfully identified the active toluene degrader Pigmentiphaga , the single cells of which were subsequently sorted and isolated by RACS. We further successfully assembled the genome of Pigmentiphaga based on the metagenomic sequencing of 13 C-DNA and genomic sequencing of sorted cells, which was confirmed by gyrB gene comparison and average nucleotide identity determination. Additionally, the genotypes and phenotypes of this degrader were directly linked at the single-cell level, and its complete toluene metabolic pathways in petroleum-contaminated soil were reconstructed. Based on its unique metabolic properties uncovered by genome sequencing, we modified the traditional cultivation medium with antibiotics, amino acids, carbon sources, and growth factors (e.g., vitamins and metals), achieving the successful cultivation of RACS-sorted active degrader Pigmentiphaga sp. Our results implied that RACS-SIP-GDC is a state-of-the-art approach for the precise identification, targeted isolation, and cultivation of functional microbes from complex communities in natural habitats. RACS-SIP-GDC can be used to explore specific and targeted organic-pollution-degrading microorganisms at the single-cell level and provide new insights into their biodegradation mechanisms.

High value food products are subject to adulterations and frauds. This study aimed to combine, in our knowledge for the first time, inorganic chemical tracers (multi-elements and Sr isotopy) with volatile organic compound (VOCs) to discriminate the geographic origin, the varieties and transformation processes to authenticate 26 tea samples. By measuring Sr isotope ratio using the multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS), 6 out of 11 regions were successfully discriminated. The combination with the ICP-MS inorganic pattern allowed to discriminate 4 more regions with a significance level of 0.05. VOCs fingerprints, obtained with selected ion flow tube mass spectrometer (SIFT-MS), were not correlated with origin but with the cultivar and transformation processes. Green, oolong, and dark teas were clearly differentiated, with hexanal and hexanol contributing to the discrimination of oxidation levels. With this multi-instrumental approach, it is possible to certify the geographical origin and the tea conformity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

This study introduces a novel methodology for utilizing historic built environments as reliable long-term geochemical archives, addressing a gap in the reconstruction of past anthropogenic pollution levels in urban settings. For the first time, we employ high-resolution laser ablation mass spectrometry for lead isotope ( 206 Pb/ 207 Pb and 208 Pb/ 206 Pb) analysis on 350-year-old black crust stratigraphies found on historic built structures, providing insights into past air pollution signatures. Our findings reveal a gradual shift in the crust stratigraphy toward lower 206 Pb/ 207 Pb and higher 208 Pb/ 206 Pb isotope ratios from the older to the younger layers, indicating changes in lead sources over time. Mass balance analysis of the isotope data shows black crust layers formed since 1669 primarily contain over 90% Pb from coal burning, while other lead sources from a set of modern pollution including but not limited to leaded gasoline (introduced after 1920) become dominant (up to 60%) from 1875 onward. In contrast to global archives such as ice cores that provide integrated signals of long-distance pollution, our study contributes to a deeper understanding of localized pollution levels, specifically in urban settings. Our approach complements multiple sources of evidence, enhancing our understanding of air pollution dynamics and trends, and the impact of human activities on urban environments.

Background: This study investigated the geographical origin classification of green coffee beans from continental to country and regional levels. An innovative approach combined stable isotope and trace element analyses with non-linear machine learning data analysis to improve coffee origin classification and marker selection. Specialty green coffee beans sourced from three continents, eight countries, and 22 regions were analyzed by measuring five isotope ratios (δ 13 C, δ 15 N, δ 18 O, δ 2 H, and δ 34 S) and 41 trace elements. Partial least squares discriminant analysis (PLS-DA) was applied to the integrated dataset for origin classification., Results: Origins were predicted well at the country level and showed promise at the regional level, with discriminating marker selection at all levels. However, PLS-DA predicted origin poorly at the continental and Central American regional levels. Non-linear machine learning techniques improved predictions and enabled the identification of a higher number of origin markers, and those that were identified were more relevant. The best predictive accuracy was found using ensemble decision trees, random forest and extreme gradient boost, with accuracies of up to 0.94 and 0.89 for continental and Central American regional models, respectively., Conclusion: The potential for advanced machine learning models to improve origin classification and the identification of relevant origin markers was demonstrated. The decision-tree-based models were superior with their embedded variable identification features and visual interpretation. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Rationale: The observed isotope distribution is an important attribute for the identification of peptides and proteins in mass spectrometry-based proteomics. Sulphur atoms have a very distinctive elemental isotope definition, and therefore, the presence of sulphur atoms has a substantial effect on the isotope distribution of biomolecules. Hence, knowledge of the number of sulphur atoms can improve the identification of peptides and proteins., Methods: In this paper, we conducted a theoretical investigation on the isotope properties of sulphur-containing peptides. We proposed a gradient boosting approach to predict the number of sulphur atoms based on the aggregated isotope distribution. We compared prediction accuracy and assessed the predictive power of the features using the mass and isotope abundance information from the first three, five and eight aggregated isotope peaks., Results: Mass features alone are not sufficient to accurately predict the number of sulphur atoms. However, we reach near-perfect prediction when we include isotope abundance features. The abundance ratios of the eighth and the seventh, the fifth and the fourth, and the third and the second aggregated isotope peaks are the most important abundance features. The mass difference between the eighth, the fifth or the third aggregated isotope peaks and the monoisotopic peak are the most predictive mass features., Conclusions: Based on the validation analysis it can be concluded that the prediction of the number of sulphur atoms based on the isotope profile fails, because the isotope ratios are not measured accurately. These results indicate that it is valuable for future instrument developments to focus more on improving spectral accuracy to measure peak intensities of higher-order isotope peaks more accurately., (© 2023 John Wiley & Sons Ltd.)

Wheat ( Triticum aestivum L.) is one of the most important cereal crops and is consumed as a staple food around the globe. Wheat authentication has become a crucial issue over the last decades. Recently, many techniques have been applied in wheat authentication including the authentication of wheat geographical origin, wheat variety, organic wheat, and wheat flour from other cereals. This paper collected related literature in the last ten years, and attempted to highlight the recent studies on the discrimination and authentication of wheat using different determination techniques and chemometric methods. The stable isotope analysis and elemental profile of wheat are promising tools to obtain information regarding the origin, and variety, and to differentiate organic from conventional farming of wheat. Image analysis, genetic parameters, and omics analysis can provide solutions for wheat variety, organic wheat, and wheat adulteration. Vibrational spectroscopy analyses, such as NIR, FTIR, and HIS, in combination with multivariate data analysis methods, such as PCA, LDA, and PLS-DA, show great potential in wheat authenticity and offer many advantages such as user-friendly, cost-effective, time-saving, and environment friendly. In conclusion, analytical techniques combining with appropriate multivariate analysis are very effective to discriminate geographical origin, cultivar classification, and adulterant detection of wheat.

Bear bile powder (BBP) is one of the most famous traditional Chinese medicines derived from animals. It has a long history of medicinal use and is widely used in the treatment of hepatobiliary and ophthalmic diseases. Due to its similar morphological characterizations and chemical composition compared with other bile powders, it is difficult to accurately identify its authenticity. In addition, there are very few methods that could analyze the geographical origins of BBP. In this study, elemental analysis isotope ratio mass spectrometry (EA-IRMS) and inductively coupled plasma mass spectrometry (ICP-MS) were used to determine stable isotope ratios and elemental contents, respectively. Combined these variables with chemometrics, the discrimination models were established successfully for identifying the authenticity and geographical origins of BBP. Meanwhile, the discrimination markers were identified by calculating the variable importance for the projection (VIP) value of each variable. A total of 13 discrimination markers (δ 13 C, δ 15 N, C, Li, Mg, K, Ca, Cr, Ni, Zn, As, Se, and Sr) were used to further establish the fingerprint of BBP. According to similarity analysis, the authenticity and geographical origins of BBP could be identified without chemometrics. In conclusion, the present study established a reliable method for authenticity identification and origin traceability of BBP, which will provide references for the quality control of bile medicines., (© 2022. Springer-Verlag GmbH Germany, part of Springer Nature.)

Geographical origin discrimination of agro-products is essential to guarantee food safety and fair trade. Garlic samples cultivated in six provinces or major production regions in China were characterized for stable isotopes (δ 13 C, δ 2 H, δ 18 O, δ 15 N, and δ 34 S), bioelemental contents (% C, % N and % S), and sulfur-containing compounds (8 organosulfur components and 2 amino acids). Results showed that many of the 18 analyzed garlic variables had significant differences among production regions. Some sulfur-containing compounds found in garlic from different provinces had a strong correlation with sulfur isotopes, suggesting garlic sulfur isotopes were also affected by geographical origin. Two supervised pattern recognition models (PLS-DA and k-NN) were developed using stable isotopes, elemental contents, and sulfur-containing compounds, and had a discrimination accuracy of 93.4 % and 87.8 %, respectively. Chemometric classification models using multi-isotopes, elements and sulfur-containing compounds provides a useful method to authenticate Chinese garlic origins., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Protein structural effects on the temperature ( T ) dependence of kinetic isotope effects (KIEs) in H-tunneling reactions have recently been used to discuss about the role of enzyme thermal motions in catalysis. Frequently observed nearly T -independent KIEs in the wild-type enzymes and T -dependent KIEs in variants suggest that H-tunneling in the former is assisted by the naturally evolved protein constructive vibrations that help sample short donor-acceptor distances (DADs) needed. This explanation that correlates the T -dependence of KIEs with DAD sampling has been highly debated as simulations following other H-tunneling models sometimes gave alternative explanations. In this paper, solvent effects on the T -dependence of KIEs of two hydride tunneling reactions of NADH/NAD + analogues (represented by Δ E a = E aD - E aH ) were determined in attempts to replicate the observations in enzymes and test the protein vibration-assisted DAD sampling concept. Effects of selected aprotic solvents on the DAD PRC 's of the productive reactant complexes (PRCs) and the DAD TRS 's of the activated tunneling ready states (TRSs) were obtained through computations and analyses of the kinetic data, including 2° KIEs, respectively. A weaker T -dependence of KIEs ( i.e. , smaller Δ E a ) was found in a more polar aprotic solvent in which the system has a shorter average DAD PRC and DAD TRS . Further results show that a charge-transfer (CT) complexation made of a stronger donor/acceptor gives rise to a smaller Δ E a . Overall, the shorter and less broadly distributed DADs resulting from the stronger CT complexation vibrations give rise to a smaller Δ E a . Our results appear to support the explanation that links the T -dependence of KIEs to the donor-acceptor rigidity in enzymes.

A flow-through cell (FTC) experiment was conducted to identify mechanisms of Ni removal by calcite through study of changes in Ni speciation and Ni isotope signature during the treatment of simulated Ni-contaminated groundwater. Synthetic Ni-contaminated groundwater was pumped through a FTC packed with crushed natural calcite. Effluent samples were collected to determine concentrations of anions, cations, and for Ni isotope-ratio measurement. X-ray absorption spectroscopy (XAS) was performed on chosen spots of the solid phase along the FTC length. Isotope data indicated multiple mechanisms affected Ni removal in the FTC system. Ni adsorption to and coprecipitation with calcite dominated the early part of the experiment yielding a fractionation factor of ε = -0.5 ‰. Subsequently, Ni precipitation as a Ni-hydroxide phase became the major process controlling Ni removal, resulting in a fractionation factor ε = -0.4 ‰. XAS analysis confirmed the presence of both Ni(OH) 2 and (Ni, Ca)CO 3 types of Ni local structural environments. Results from this study highlight the potential of Ni isotopes as auxiliary tools to determine the processes involved in Ni attenuation from the environment. The characterization of mechanisms involved in Ni removal from solution is necessary to evaluate potential impacts to the environment and to develop effective remediation strategies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

We present a fast, accurate, and robust approach for determination of free energy profiles and kinetic isotope effects for RNA 2'-O-transphosphorylation reactions with inclusion of nuclear quantum effects. We apply a deep potential range correction (DPRc) for combined quantum mechanical/molecular mechanical (QM/MM) simulations of reactions in the condensed phase. The method uses the second-order density-functional tight-binding method (DFTB2) as a fast, approximate base QM model. The DPRc model modifies the DFTB2 QM interactions and applies short-range corrections to the QM/MM interactions to reproduce ab initio DFT (PBE0/6-31G*) QM/MM energies and forces. The DPRc thus enables both QM and QM/MM interactions to be tuned to high accuracy, and the QM/MM corrections are designed to smoothly vanish at a specified cutoff boundary (6 Å in the present work). The computational speed-up afforded by the QM/MM+DPRc model enables free energy profiles to be calculated that include rigorous long-range QM/MM interactions under periodic boundary conditions and nuclear quantum effects through a path integral approach using a new interface between the AMBER and i-PI software. The approach is demonstrated through the calculation of free energy profiles of a native RNA cleavage model reaction and reactions involving thio-substitutions, which are important experimental probes of the mechanism. The DFTB2+DPRc QM/MM free energy surfaces agree very closely with the PBE0/6-31G* QM/MM results, and it is vastly superior to the DFTB2 QM/MM surfaces with and without weighted thermodynamic perturbation corrections. 18 O and 34 S primary kinetic isotope effects are compared, and the influence of nuclear quantum effects on the free energy profiles is examined.

Isotope substitution, a fascinating tool of physical chemistry, has been broadly applied in the research field of heterogeneous catalysis. In general, due to the differences in the mass-related atomic vibrational frequencies and zero-point energy of isotopic molecules, the apparent isotope effect (AIE) or observed kinetic isotope effect (observed KIE) from isotope substitution examination could provide unique knowledge regarding the reaction rate and mechanism of a catalytic reaction, such as the rate-determining step, key reaction intermediate, or catalyst design and synthesis. However, the treatment of the AIE is not as straightforward as the isotopic switch experiment, and needs sufficient care and comprehensive identification to deal with the influences from the equilibrium isotope effects (EIEs) of quasi-equilibrium elementary steps, kinetic isotope effect (KIE) of the pseudo rate-determining step, transition states, intrinsic reaction barriers, etc. Fundamentally, the key factors affecting the AIE could be the partition function part and the zero-point energy part of each single elementary step. Theoretically, the classification of the KIE could be based on the quantity of KIE (including normal KIE and inverse KIE) or the molecular transformation (including primary KIE, secondary KIE, tunneling KIE, and solvent KIE) involved. This article presents a recap of the fundamental concepts and relations of KIE, EIE and AIE, and a concise review on the selected applications of isotope effects throughout heterogeneous catalysis. Lastly, the meaningful perspectives regarding the critical factors that impact the AIE and the appropriate treatment of the AIE are discussed meticulously.

Background: Flaxseed is an economically important oilseed crop whose geographic origin is of significant interest to producers and consumers because every region may exhibit particular quality characteristics. The lipid/fatty acid method of determining the geographic origin of flaxseed has not been found to be adequate., Results: To improve the discrimination rate and the geographical traceability of this crop, the chemical profiles of the flaxseed samples were characterized via lipids/fatty acids, stable isotopes, and antioxidant capacity. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were also performed. A satisfactory discrimination rate of 98.6% was obtained after combining fatty acids, stable isotopes, and antioxidant capacity to trace the origin of flaxseed from five regions in northern China., Conclusion: This study provides an effective method for distinguishing the geographic origin of flaxseed. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

The reported method involves a novel workflow that eliminates the need for authentic reference standards for the quantitation of drug metabolites in biological samples using a single multi-isotopically labeled compound bearing both radio and stable isotopes. The resulting radio and stable bifunctionalized isotopolog (RADSTIL) of the parent drug is employed as a substrate for in vitro biotransformation to targeted RADSTILs of metabolites as calibrants. Inclusion of a radio label enables both radiometric and mass spectrometric detection. The addition of stable labels ensures the subsequent isotopic interference-free quantitation of unlabeled metabolites in preclinical and clinical samples. This affords a more accurate quantitation workflow compared with the current semi-quantitation method, which utilizes isotopic interfering radio isotopologs of metabolites alone as calibrants. The proof-of-concept is illustrated with ( 14 C, 13 C 2 )-acetaminophen where in vitro biotransformation produced ( 14 C, 13 C 2 )-sulfate and ( 14 C, 13 C 2 )-glucuronide calibrants. Absolute quantitation of the acetaminophen metabolites was then achieved by liquid chromatography coupled with radiometry and mass spectrometry. Quantitative data obtained by this method fell within 82-86% of the values from conventional LC-MS/MS method., (© 2021 John Wiley & Sons, Ltd.)

Soil microorganisms determine the fate of soil organic matter (SOM), and their activities compose a major component of the global carbon (C) cycle. We employed a multisubstrate, DNA-stable isotope probing experiment to track bacterial assimilation of C derived from distinct sources that varied in bioavailability. This approach allowed us to measure microbial contributions to SOM processing by measuring the C assimilation dynamics of diverse microorganisms as they interacted within soil. We identified and tracked 1,286 bacterial taxa that assimilated 13 C in an agricultural soil over a period of 48 d. Overall 13 C-assimilation dynamics of bacterial taxa, defined by the source and timing of the 13 C they assimilated, exhibited low phylogenetic conservation. We identified bacterial guilds composed of taxa that had similar 13 C assimilation dynamics. We show that C-source bioavailability explained significant variation in both C mineralization dynamics and guild structure, and that the growth dynamics of bacterial guilds differed significantly in response to C addition. We also demonstrate that the guild structure explains significant variation in the biogeographical distribution of bacteria at continental and global scales. These results suggest that an understanding of in situ growth dynamics is essential for understanding microbial contributions to soil C cycling. We interpret these findings in the context of bacterial life history strategies and their relationship to terrestrial C cycling., Competing Interests: The authors declare no competing interest.

The Late Triassic Carnian Pluvial Episode (CPE) saw a dramatic increase in global humidity and temperature that has been linked to the large-scale volcanism of the Wrangellia large igneous province. The climatic changes coincide with a major biological turnover on land that included the ascent of the dinosaurs and the origin of modern conifers. However, linking the disparate cause and effects of the CPE has yet to be achieved because of the lack of a detailed terrestrial record of these events. Here, we present a multidisciplinary record of volcanism and environmental change from an expanded Carnian lake succession of the Jiyuan Basin, North China. New U-Pb zircon dating, high-resolution chemostratigraphy, and palynological and sedimentological data reveal that terrestrial conditions in the region were in remarkable lockstep with the large-scale volcanism. Using the sedimentary mercury record as a proxy for eruptions reveals four discrete episodes during the CPE interval (ca. 234.0 to 232.4 Ma). Each eruptive phase correlated with large, negative C isotope excursions and major climatic changes to more humid conditions (marked by increased importance of hygrophytic plants), lake expansion, and eutrophication. Our results show that large igneous province eruptions can occur in multiple, discrete pulses, rather than showing a simple acme-and-decline history, and demonstrate their powerful ability to alter the global C cycle, cause climate change, and drive macroevolution, at least in the Triassic., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Background: The use of PDO (protected designation of origin) and PGI (protected geographical indication) labels allows to protect and promote agricultural products characterized by unique features related to the place of origin and traditional know-how. However, the presence of non-authentic products in the market represents a fraud that can be tackled applying analytical techniques combined with chemometric analysis. In this study, we applied multi-element and multi-isotope analysis to characterize PDO and PGI apples cultivated in northern Italy, comparing them with Italian apples without labels of geographical indications., Results: The multi-element and multi-isotope approach allowed to characterize the apples cultivated in northern Italy. Despite a significant effect of the sampling sites on the apple composition, the comparison of the multi-chemical fingerprint of the apples significantly varied among cultivation areas. Results of this characterization were used to classify samples according to their cultivation area applying a linear discriminant analysis (LDA). Outputs of the LDA showed that correct sample classification can be successfully achieved (balanced accuracy > 96%). Moreover, using a selection of variables, it was possible to correctly classify samples also at regional level., Conclusion: The presented evidences indicate that the multi-element and multi-isotope fingerprint can be successfully applied to traceability studies. The combination of this characterization with chemometric tools allows the classification of Italian apples based on their origin both on a national and regional scale. This approach represents an interesting tool to enhance and protect PDO and PGI Italian products. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Rationale: The study of insect migration is problematic due to the small size of insects. Stable isotope analysis can be used to elucidate movement, either by geographic assignment of location of a species, or by simply distinguishing migrant from resident populations. There are few isoscapes of any kind in the UK/Ireland available for interrogation. Thus, I have measured stable isotope ratios (of H, C, N and S) of 299 individuals of the non-migratory Brimstone moth (Opisthograptis luteolata) collected from 93 locations around the UK and Ireland by citizen scientists., Methods: After removing lipids, stable isotope ratios were measured by continuous flow isotope ratio mass spectrometry, using either a conventional elemental analyser (C, N and S) or a high-temperature, thermal conversion elemental analyser in reductive mode., Results: Maps (isoscapes) were constructed that illustrate the stable isotope spatial distribution of this insect. These are the first isoscapes of H, C, N and S of biological samples covering both UK and Ireland., Conclusions: The insect isoscape patterns can be explained from what we know of moth diet, climate and geology. Sulfur isotopes may be of particular use for distinguishing individuals from areas of unique geology. Isoscape patterns may (with care) predict isotope compositions of other, herbivorous, non-aquatic, chitinous taxa. Such isoscapes, when extended beyond the UK and Ireland, would provide a useful tool to elucidate insect migration., (© 2021 The Author. Rapid Communications in Mass Spectrometry published by John Wiley & Sons Ltd.)

Boron neutron capture therapy (BNCT) is a two-step therapeutic process that utilizes Boron-10 in combination with low energy neutrons to effectively eliminate targeted cells. This therapy is primarily used for difficult to treat head and neck carcinomas; recent advances have expanded this method to cover a broader range of carcinomas. However, it still remains an unconventional therapy where one of the barriers for widespread adoption is the adequate delivery of Boron-10 to target cells. In an effort to address this issue, we examined a unique nanoparticle drug delivery system based on a highly stable and modular proteinaceous nanotube. Initially, we confirmed and structurally analyzed ortho-carborane binding into the cavities of the nanotube. The high ratio of Boron to proteinaceous mass and excellent thermal stability suggest the nanotube system as a suitable candidate for drug delivery into cancer cells. The full physicochemical characterization of the nanotube then allowed for further mechanistic molecular dynamic studies of the ortho-carborane uptake and calculations of corresponding energy profiles. Visualization of the binding event highlighted the protein dynamics and the importance of the interhelical channel formation to allow movement of the boron cluster into the nanotube. Additionally, cell assays showed that the nanotube can penetrate outer membranes of cancer cells followed by localization around the cells' nuclei. This work uses an integrative approach combining experimental data from structural, molecular dynamics simulations and biological experiments to thoroughly present an alternative drug delivery device for BNCT which offers additional benefits over current delivery methods., (© 2021. The Author(s).)

A hierarchical hollow Ni/Co-codoped MoS2 architecture was successfully prepared using a Ni/Co Prussian Blue analogue as the precursor followed by the solvothermal-assisted insertion of MoS42- and extraction of [Co(CN)6]3- at 200 °C for 32 h. The obtained Ni/Co-codoped MoS2 composite exhibited a hollow microcubic structural characteristic, and the morphology, structure, and chemical compositions were carefully characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS), respectively. The Ni/Co-codoped MoS2 composite used as an electrode material featured excellent glucose sensing activity and a high sensitivity of 2546 μA mM-1 cm-2 with a relatively low detection limit of 0.69 μM (S/N = 3). In addition, the Ni/Co-codoped MoS2 composite showed good anti-interference sensing performance in the presence of ascorbic acid (AA), lysine (Lys), cysteine (Cys), urea, H2O2, KCl, and other interferents. These experimental results revealed that the composite is a promising electrode material for enzyme-free glucose sensing, and the feasible synthetic strategy may provide an effective and controlled route to prepare other multi-metal substituted sulfide-based hierarchical structures with high electrochemical sensing performance.

The increasing number of available genomes, in combination with advanced genome mining techniques, unveiled a plethora of biosynthetic gene clusters (BGCs) coding for ribosomally synthesized and post-translationally modified peptides (RiPPs). The products of these BGCs often represent an enormous resource for new and bioactive compounds, but frequently, they cannot be readily isolated and remain cryptic. Here, we describe a tunable metabologenomic approach that recruits a synergism of bioinformatics in tandem with isotope- and NMR-guided platform to identify the product of an orphan RiPP gene cluster in the genomes of Nocardia terpenica IFM 0406 and 0706 T . The application of this tactic resulted in the discovery of nocathioamides family as a founder of a new class of chimeric lanthipeptides I., (© 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

The stable isotopic composition of diatom silica is used as a proxy for nutrient utilisation in natural waters. This approach provides essential insight into the current and historic links between biological production, carbon cycling and climate. However, estimates of isotopic fractionation during diatom silica production from both laboratory and field studies are variable, and the biochemical pathways responsible remain unknown. Here, we investigate silicon isotopic fractionation through a series of chemical precipitation experiments that are analogous to the first stages of intracellular silica formation within the diatom silicon deposition vesicle. The novelty of our experiment is the inclusion of the R5 peptide, which is closely related to a natural biomolecule known to play a role in diatom silicification. Our results suggest that the presence of R5 induces a systematic but non-significant difference in fractionation behaviour. It thus appears that silicon isotopic fractionation in vitro is largely driven by an early kinetic fractionation during rapid precipitation that correlates with the initial amount of dissolved silica in the system. Our findings raise the question of how environmental changes might impact silicon isotopic fractionation in diatoms, and whether frustule archives record information in addition to silica consumption in surface water.

In this study, stable isotopes and multi-element signatures combined with chemometrics were used to distinguish conventional and organic Chinese yams based on field trials. Four light stable isotopes δD, δ 13 C, δ 15 N, δ 18 O, and 20 elements (e.g. Li, Na, Mn) were determined, then evaluated using significance analysis and correlation analysis, and modeling of various chemometrics methods. Consequently, the RandomForest model showed the best performance with AUC value of 0.972 and predictive accuracy of 97.3%, and Mn, Cr, Se, Na, δD, As, and δ 15 N were screened as significant variables. Moreover, many chemical components and antioxidant activity of yam samples were determined spectrophotometrically. The results indicated that organic yams had advantages in secondary metabolites such as polyphenol, flavonoid and saponin; conversely, conventional samples had more primary metabolites like protein and amino acids. Above all, this work provides a beneficial case in the authentication and quality evaluation of conventional and organic yams., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A new method combining isotope dilution mass spectrometry (IDMS) and standard addition has been developed to determine the mass fractions w of different elements in complex matrices: (a) silicon in aqueous tetramethylammonium hydroxide (TMAH), (b) sulfur in biodiesel fuel, and (c) iron bound to transferrin in human serum. All measurements were carried out using inductively coupled plasma mass spectrometry (ICP-MS). The method requires the gravimetric preparation of several blends (b i )-each consisting of roughly the same masses ( m x, i ) of the sample solution (x) and m y, i of a spike solution (y) plus different masses ( m z, i ) of a reference solution (z). Only these masses and the isotope ratios ( R b, i ) in the blends and reference and spike solutions have to be measured. The derivation of the underlying equations based on linear regression is presented and compared to a related concept reported by Pagliano and Meija. The uncertainties achievable, e.g., in the case of the Si blank in extremely pure TMAH of u rel ( w (Si)) = 90% (linear regression method, this work) and u rel ( w (Si)) = 150% (the method reported by Pagliano and Meija) seem to suggest better applicability of the new method in practical use due to the higher robustness of regression analysis.

Background: The present COVID-19 pandemic has prompted worldwide repurposing of drugs. The aim of the present work was to develop and validate a two-dimensional isotope-dilution liquid chromatrography tandem mass spectrometry (ID-LC-MS/MS) method for accurate quantification of remdesivir and its active metabolite GS-441524, chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin in serum; drugs that have gained attention for repurposing in the treatment of COVID-19., Methods: Following protein precipitation, samples were separated with a two-dimensional ultra-high performance liquid chromatography (2D-UHPLC) setup, consisting of an online solid phase extraction (SPE) coupled to an analytical column. For quantification, stable isotope-labelled analogues were used as internal standards for all analytes. The method was validated on the basis of the European Medicines Agency bioanalytical method validation protocol., Results: Detuning of lopinavir and ritonavir allowed simultaneous quantification of all analytes with different concentration ranges and sensitivity with a uniform injection volume of 5 μL. The method provided robust validation results with inaccuracy and imprecision values of ≤ 9.59 % and ≤ 11.1 % for all quality controls., Conclusion: The presented method is suitable for accurate and simultaneous quantification of remdesivir, its metabolite GS-441525, chloroquine, hydroxychloroquine, lopinavir, ritonavir, favipiravir and azithromycin in human serum. The quantitative assay may be an efficient tool for the therapeutic drug monitoring of these potential drug candidates in COVID-19 patients in order to increase treatment efficacy and safety., Competing Interests: Declaration of Competing Interest The authors have no conflict of interest to declare., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A routine, selective and sensitive ultra-high performance liquid chromatography-electrospray ionisation tandem triple quadrupole mass spectrometry (UHPLC-ESI-MS/MS) method was developed and validated for the quantification of 3-monochloropropane-1,2-diol (3-MCPD) in Caco-2 cell transport buffer (FaSSIF-V2, the second version of a fasted state simulated intestinal fluid) and receiving buffer (HBSS, Hank's balanced salt solution). The method involves measuring deuterated 3-MCPD (3-MCPD- d5 ) as internal standard (IS) during the entire analytical procedure to obtain precise and accurate results. The separation was performed on a Poroshell 120 HILIC column (2.7  µ m, 3.0 × 50 mm) at a flow rate of 0.3 mL/min using water (containing 0.025% acetic acid) and acetonitrile (containing 0.025% acetic acid) as the mobile phases. Mass spectrometric detection was operated in dynamic multiple reaction monitoring (dMRM) in negative ion mode. The method exhibited high sensitivity. The limits of detection (LOD) for 3-MCPD in FaSSIF-V2 and HBSS were 0.012 and 0.014  µ mol/L, and the limits of quantification (LOQ) were 0.039 and 0.045  µ mol/L, respectively. Satisfactory results were observed for linearity ( R 2   > 0.999), intra-day precision ( RSD % <7.7% in FaSSIF-V2 and <6.6% in HBSS), inter-day precision ( RSD % <5.9% in FaSSIF-V2 and <5.6% in HBSS), accuracy (% error within ± 10%), and sample stability ( RSD % <7.7% and % error within ± 10%). The method has been successfully applied to quantify 3-MCPD in Caco-2 cell transport and receiving buffers. The results were in good agreement with those obtained with gas chromatography-tandem mass spectrometry (GC-MS).

The most important task in the design of dosage forms is to modify the pharmaceutical substances structure in order to increase solubilization, targeted delivery, controlled rate of drug administration, and its bioavailability. Screening-laboratory (in vitro) or computer (in silico)-as a procedure for selecting a prototype for the design of a drug molecule, involves several years of research and significant costs. Among a large number of solvents and diluents (alcohol, ether, oils, glycerol, Vaseline) used in the pharmaceutical industry for the manufacture of drugs water finds the greatest application. This is because all biological reactions (reactions in living systems) take place in water and distribution of the fluid in the body and the substances found within is critical for the maintenance of intracellular and extracellular functions. Modern studies in the field of the stable isotopic compositions of natural water and its structure and properties make it possible to use isotopic transformations of the water to improve the pharmacokinetic properties of medicinal substances without previous structural modification. It is known that by replacing any of the atoms in the reacting substance molecule with its isotope, it is possible to record changes in the reactivity, which are expressed as a change in the reaction rate constant, i.e., in the manifestation of the kinetic isotope effect (KIE). The article presents the results of studies on the effect of the kinetic isotope effect of a solvent-water-on increasing the solubility and dissolution rate constants of poorly soluble drugs using laser diffraction spectroscopy. The results of the studies can be successfully implemented in pharmaceutical practice to overcome the poor solubility of medicinal substances of classes II and IV, according to the biopharmaceutical classification system (BCS), in water for pharmaceutical purposes by performing its preliminary and safe isotopic modification.

Oxygen, hydrogen, carbon and nitrogen stable isotopes were measured on a comprehensive sampling of feathers from two spring Hooded Warblers (Setophaga citrina) in Texas to evaluate isotopic variability between feathers and during molt. Isotopic homogeneity within each bird was found across all four isotopic systems, supporting the hypothesis that molt in these neotropical migrants is fully completed on the breeding grounds. This homogeneity suggests that the isotopic composition of a single feather is may be representative of the whole songbird. However, each bird was found to have one or two outlier feathers, which could signify regrowth of lost feathers after prebasic molt., Competing Interests: The authors have declared that no competing interests exist.

A compound-specific chlorine/bromine isotope analysis (Cl-/Br-CSIA) method was developed using gas chromatography-quadrupole mass spectrometry for polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), which are toxic to human health and are frequently detected in various abiotic and biotic media. For PCB congeners, the molecular ion method for a concentration of 0.5-10.0 ppm, a dwell time of 20-100 ms, a relative EM voltage of 200 V, an electric current of 34 μA, and an ionization energy of 70 eV was determined as the most suitable scheme, which obtained standard deviations (SDs) of chlorine isotope ratios ranging from 0.00008 to 0.00068. As for the PBDE congeners, the lowest SDs, ranging from 0.00050 to 0.00172, were determined using the top four ion method with a concentration of 5-10 ppm and a dwell time of 20-50 ms. Both the chlorine and bromine isotope ratios showed strong concentration dependencies. Therefore, external standardization or detecting chlorine and bromine isotope ratios at a uniform concentration level is necessary to eliminate the concentration effect. In addition, 13 C-correction is critical to remove interference from carbon isotopes. This newly developed Cl-/Br-CSIA method successfully determined the chlorine/bromine isotope ratios of PCBs/PBDEs in technical mixtures and traced the chlorine/bromine isotope ratio variations of PCBs/PBDEs in photodegradation experiments, thereby suggesting that it is a promising tool for assessing the sources and transformation processes of PCBs and PDBEs in the environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Food authenticity concerning the geographical origin becomes increasingly important for consumers, food industries, and food authorities. In this study, nontargeted 1 H NMR metabolomics combined with machine learning methodologies was applied to successfully distinguish the geographical origin of 237 samples of white asparagus from Germany, Poland, The Netherlands, Spain, Greece, and Peru. Support vector classification of the geographical origin achieved an accuracy of 91.5% for the entire sample set and 87.8% after undersampling the majority class. Important regions of the spectra could be identified and assigned to potential chemical markers. A subset of samples was compared to isotope-ratio mass spectrometry (IRMS), an established method for the determination of origin of white asparagus in Germany. Here, SVM classification led to accuracies of 79.4% for NMR and 70.9% for IRMS. Finally, the classification of asparagus from different German regions was evaluated, and the influence of year and variety was analyzed.

Introduction: The most common and aggressive primary malignancy of the central nervous system is Glioblastoma that, as a wide range of malignant solid tumor, is characterized by extensive hypoxic regions. A great number of PET radiopharmaceuticals have been developed for the identification of hypoxia in solid tumors, among these, we find copper-based tracers. The aim of the current review paper was to provide an overview of radiocopper compounds applied for preclinical and clinical research in brain tumors and hypoxia imaging or therapy., Evidence Acquisition: Copper offers a wide variety of isotopes, useful for nuclear medicine applications, but only 64 Cu and 67 Cu are under the spotlight of the scientific community since being good candidates for theranostic applications. Between the two, 64 Cu availability and production cost have attracted more interest of the scientific community., Evidence Synthesis: In order to better understand the application of copper-bis thiosemicarbazones in hypoxia imaging, an overview of the role of hypoxia in cancer, existing non-imaging and imaging techniques for hypoxia identification and promising future avenues regarding hypoxia is necessary. Different proposed uptake mechanisms of [ 64 Cu][Cu(ATSM)] inside the cell will be discussed and other 64 Cu-based tracers for brain tumors described., Conclusions: Among radio copper compounds [ 64 Cu][Cu(ATSM)] is the most studied radiopharmaceutical for imaging and treatment of brain tumors. Experimental evidence suggested that [ 64 Cu][Cu(ATSM)] could be more appropriately considered as a marker of over-reduced intracellular state rather than a pure hypoxia agent. Moreover, preliminary clinical data suggested that [ 64 Cu]CuCl2 can be a potentially useful diagnostic agent for malignancies of the central nervous system (CNS).

The shortcomings in Boron neutron capture therapy (BNCT) and Hyperthermia for killing the tumor cell desired for the synthesis of a new kind of material suitable to be first used in BNCT and later on enable the conditions for Hyperthermia to destroy the tumor cell. The desire led to the synthesis of large band gap semiconductor nano-size Boron-10 enriched crystals of hexagonal boron nitride ( 10 BNNCs). The contents of 10 BNNCs are analyzed with the help of x-ray photoelectron spectroscopy (XPS) and counter checked with Raman and XRD. The 10 B-contents in 10 BNNCs produce 7 Li and 4 He nuclei. A Part of the 7 Li and 4 He particles released in the cell is allowed to kill the tumor (via BNCT) whereas the rest produce electron-hole pairs in the semiconductor layer of 10 BNNCs suggested to work in Hyperthermia with an externally applied field., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Magnesium isotope ratios (26Mg/24Mg) can provide insights into the origin of Mg pools and fluxes in catchments where Mg sources have distinct isotope compositions, and the direction and magnitude of Mg isotope fractionations are known. Variability in Mg isotope compositions was investigated in three small, spruce-forested catchments in the Czech Republic (Central Europe) situated along an industrial pollution gradient. The following combinations of catchment characteristics were selected for the study: low-Mg bedrock + low Mg deposition (site LYS, underlain by leucogranite); high-Mg bedrock + low Mg deposition (site PLB, underlain by serpentinite), and low-Mg bedrock + high Mg deposition (site UDL, underlain by orthogneiss). UDL, affected by spruce die-back due to acid rain, was the only investigated site where dolomite was applied to mitigate forest decline. The δ26Mg values of 10 catchment compartments were determined on pooled subsamples. At LYS, a wide range of δ26Mg values was observed across the compartments, from -3.38 ‰ (bedrock) to -2.88 ‰ (soil), -1.48% (open-area precipitation), -1.34 ‰ (throughfall), -1.19 ‰ (soil water), -0.99 ‰ (xylem), -0.95 ‰ (needles), -0.82 ‰ (bark), -0.76 ‰ (fine roots), and -0.76 ‰ (runoff). The δ26Mg values at UDL spanned 1.32 ‰ and were thus less variable, compared to LYS. Magnesium at PLB was isotopically relatively homogeneous. The δ26Mg systematics was consistent with geogenic control of runoff Mg at PLB. Mainly atmospheric/biological control of runoff Mg was indicated at UDL, and possibly also at LYS. Our sites did not exhibit the combination of low-δ26Mg runoff and high-δ26Mg weathering products (secondary clay minerals) reported from several previously studied sites. Six years after the end of liming at UDL, Mg derived from dolomite was isotopically undetectable in runoff., Competing Interests: The authors have declared that no competing interests exist.

N -heterocyclic carbenes (NHCs) are common ancillary ligands in organometallic compounds that are used to alter the electronic and steric properties of a metal centre. To date, various NHCs have been synthesised with different electronic properties, which can be done by modifying the backbone or changing the nitrogen substituents group. This study describes a systematic modification of NHCs by the inclusion of fluorine substituents and examines the use of selenium-NHC compounds to measure the π-accepting ability of these fluorinated NHC ligands. Evaluation of the 77 Se NMR chemical shifts of the selenium adducts reveals that fluorinated NHCs have higher chemical shifts than the non-fluorinated counterparts, IMes and IPh. Higher 77 Se NMR chemical shifts values indicate a stronger π-accepting ability of the NHC ligands. The findings of this study suggest that the presence of fluorine atoms has increased the π-accepting ability of the corresponding NHC ligands. This work supports the advantage of the 77 Se NMR chemical shifts of selenium-NHC compounds for assessing the influence of fluorine substituents on NHC ligands.

Cyclic RGD (cRGD) peptide-conjugated boronated albumin was developed to direct toward integrin α v β 3, which overexpresses on many cancer cells. A stepwise conjugation of c[RGDfK(Mal)] and maleimide-conjugated closo -dodecaborate (MID) to bovine serum albumin (BSA) afforded cRGD-MID-BSA, which was noncytotoxic toward both U87MG and A549 cells. As compared with l-BPA, selective antitumor activity of cRGD-MID-BSA toward U87MG cells overexpressing integrin α v β 3 was identified after thermal neutron irradiation. In vivo fluorescence live imaging of Cy5-conjugated cRGD-MID-BSA and MID-BSA revealed that both cRGD-MID-BSA and MID-BSA similarly reached the maximum accumulation during 8-12 h after injection. The selective accumulation and retention of Cy5-cRGD-MID-BSA was more pronounced than Cy5-MID-BSA after 24 h. An in vivo boron neutron capture therapy (BNCT) study revealed that the cRGD peptide ligand combination enhanced accumulation of MID-BSA into tumor cells in U87MG xenograft models. The significant tumor growth suppression was observed in U87MG xenograft models at a dose of 7.5 mg [ 10 B]/kg after neutron irradiation.

Vicinal diols and its derivatives can be exploited as model compounds for the investigation of radiation-induced free-radical transformations of hydroxyl-containing biomolecules such as carbohydrates, phospholipids, ribonucleotides, amino acids, and peptides. In this paper, for the first time, the prospects of isotope reinforcement approach in inhibiting free-radical transformations of hydroxyl-containing compounds in aqueous solutions are investigated on the example of radiolysis of 1,2-propanediol and 1,2-propanediol-2-d 1 aqueous solutions. At an absorbed dose rate of 0.110 ± 0.003 Gy·s -1 a profound kinetic isotope effect (KIE) is observed for the non-branched chain formation of acetone, which is a final dehydration product of predominant carbon-centred radicals CH 3 ·C(OH)CH 2 OH. In 0.1 and 1 M deaerated solutions at pH 7.00 ± 0.01, the values of KIE are 8.9 ± 1.7 and 15.3 ± 3.1, respectively. A rationale for the fact that a strong KIE takes place only in the case of chain processes, which may occur during free-radical transformations of vicinal diols, is also provided herein based on the results of 2-propanol and 2-propanol-2-d 1 indirect radiolysis. Lastly, the lack of KIE is shown in the case of 2-butanone formation from 2,3-butanediol or 2,3-butanediol-2,3-d 2 . This indicates that the type (primary, secondary) of the β-carbonyl radicals formed as a result of CH 3 ·C(OH)CH(OH)R (R = H, CH 3 ) dehydration determines the manifestation of the effect.

Vitamin B 6 comprises an important set of molecules tightly interwoven with the human amino acid, fatty acid, and carbohydrate metabolism. Analytical methods striving for the quantification of individual B 6 vitamers so far mostly rely on methods based on HPLC in combination with fluorescence detection, but their application encounters multiple difficulties due to the chemical divergence of the single vitamers. The present study describes the development of a method based on LC-MS/MS and stable isotope dilution assay (SIDA) for the simultaneous quantification of five vitamers (PN, PL, PM, PMP, and PNG) of the B 6 group in food samples. [ 13 C 3 ]-PN, [ 13 C 3 ]-PL, and [ 13 C 6 ]-PNG were applied as internal standards for the analysis of PN, PL, and PNG. PM and PMP were quantified via matrix-matched calibration referring to [ 13 C 3 ]-PN. The developed method was validated using starch matrix. The limits of detection and quantification ranged from 0.0028 to 0.02 mg/kg and from 0.0085 to 0.059 mg/kg, respectively, for all analytes. Calculated recoveries varied from 92 to 111%. Intra-injection precisions ranged from 0 to 9%, inter-day precisions from 4 to 10%, and intra-day precisions from 4 to 10%. A total of 14 plant-based food samples including fruits, vegetables, and cereals were examined for their content of vitamin B 6 using the validated method. Furthermore, the first quantitation of PNG without enzymatic steps or divergent internal standards was undertaken utilizing LC-MS/MS and SIDA.

This review deals with biological systems and with deuterium isotope effects on chemical shifts caused by the replacement of OH, NH or SH protons by deuterons. Hydrogen bonding is clearly of central importance. Isotope effects on chemical shifts seems very suitable for use in studies of structures and reactions in the interior of proteins, as exchange of the label can be expected to be slow. One-bond deuterium isotope effects on 15 N chemical shifts, and two-bond effects on 1 H chemical shifts for N(D)H x systems can be used to gauge hydrogen bond strength in proteins as well as in salt bridges. Solvent isotope effects on 19 F chemical shifts show promise in monitoring solvent access. Equilibrium isotope effects need in some cases to be taken into account. Schemes for calculation of deuterium isotope effects on chemical shifts are discussed and it is demonstrated how calculations may be used in the study of complex biological systems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The early evidence of domesticated animals and human-animal interaction in South Asia can be traced back to the seventh millennium BCE; however, our understanding of their use is incomplete and limited to the analysis of animal bones from archaeological sites. By the third millennium BCE with the emergence of the Indus Civilization, cattle and water-buffalo became the primary domesticates and outnumbered any other animals at the majority of the Indus settlements. Based on the analysis of skeletal remains and ethnographic data, a number of studies have suggested that cattle and water-buffalo were utilized for their meat, dairy, hides, and other labor-oriented jobs. While some of these claims are backed by empirical data, others are primarily discussed as hypotheses, for example, the exploitation of dairy. In this paper, by analyzing the absorbed lipid residues from fifty-nine ceramic sherds recovered from an agro-pastoral settlement that was occupied during the peak of the Indus period around mid- to late third millennium BCE, we provide the earliest direct evidence of dairy product processing, particularly from cattle and possibly from some water-buffalo. By providing direct evidence of animal product processing, we identify the use of primary domesticated animals and other resources in the diet during the Indus Civilization.

Extending oceanographic data beyond the instrumental period is highly needed to better characterize and understand multi-decadal to centennial natural ocean variability. Here, a stable isotope record at unprecedented temporal resolution (1 to 2 years) from a new marine core retrieved off western North Iceland is presented. We aim to better constrain the variability of subsurface, Atlantic-derived Subpolar Mode Water (SPMW), using near surface-dwelling planktic foraminifera and Arctic Intermediate Water (AIW) mass changes using benthic foraminifera over the last ~165 years. The reconstruction overlaps in time with instrumental observations and a direct comparison reveals that the δ18O record of Neogloboquadrina pachyderma is reliably representing temperature fluctuations in the SPMWs. Trends in the N. pachyderma δ13C record match the measured phosphate concentration in the upper 200 m on the North Icelandic Shelf well. Near surface-dwelling foraminifera trace anthropogenic CO2 in the Iceland Sea by ~ 1950 ± 8, however, a reduced amplitude shift in the Marine Suess effect is identified. We argue that this is caused by a contemporary ongoing increase in marine primary productivity in the upper ocean due to enhanced Greenland's freshwater discharge that has contributed to a nutrient-driven fertilization since the 1940s/50s (Perner et al., 2019). Multi-decadal variability is detected. We find that the 16-year periodicity evident in SPMW and AIWs based on the δ18O of N. pachyderma and M. barleeanum is a signal of SST anomalies propagated into the Nordic Seas via the Atlantic inflow branches around Iceland. Spectral analyses of the planktic foraminiferal δ13C signal indicate intermittent 30-year cycles that are likely reflecting the ocean response to atmospheric variability, presumably the East Atlantic Pattern. A long-term trend in benthic δ18O suggests that Atlantic-derived waters are expanding their core within the water column from the subsurface into deeper intermediate depths towards the present day. This is a result of increased transport by the North Icelandic Irminger Current to the North Iceland Shelf over the historical era., Competing Interests: The authors have declared that no competing interests exist.

Theoretical models aimed at describing magic-angle-spinning (MAS) dynamic nuclear polarization (DNP) NMR have great potential in facilitating the in silico design of DNP polarizing agents and formulations. These models must typically face a trade-off between the accuracy of a strict quantum mechanical description and the need for using realistically large spin systems, for instance, using phenomenological models. Here, we show that the use of aggressive state-space restrictions and an optimization strategy allows full-scale ab initio MAS-DNP simulations of spin systems containing thousands of nuclei. Our simulations are shown to reproduce experimental DNP enhancements quantitatively, including their MAS rate dependence, for both frozen solutions and solid materials. They also reveal the importance of a previously unrecognized structural feature found in some polarizing agents that helps minimize the sensitivity losses imposed by the spin diffusion barrier.

Aim: To optimize clinical efficacy and reduce the drug-exposure-related toxicity of linezolid, whose concentrations show wide inter-variabilities, a simple and reliable quantitative assay for therapeutic drug monitoring is necessary. Results: A UHPLC-MS/MS assay has been established for determination of linezolid in human plasma and fully validated according to the US FDA guidelines. After a simple, isotope-dilluted precipitation with methanol, the analytes were separated by a straightforward isocratic mode and the MS/MS was conducted under the ESI + mode fitted with SRM. The calibration curves proved acceptable linearity in the range of 0.1-30.0 µg/ml. Conclusion: The present assay is currently used in routine clinical practice, being applied to therapeutic drug monitoring and helps to optimize individual dosing regimens and manage adverse effects in ICU patients.

Dual-mode heart-cutting two-dimensional liquid chromatography (DMHC 2D-LC) was applied to isotope dilution mass spectrometry (IDMS) to reduce the bias in the quantitative analysis of a target analyte present in a limited quantity in human plasma. Based on a Waters I-Class LC system, the DMHC 2D-LC system was operated in one- and two-dimensional modes to facilitate the determination of heart-cutting time and the efficient trapping of the target LC eluate. Experiments to determine the feasibility of coupling with IDMS were performed with triple quadrupole mass spectrometry using folic acid standards and/or 13 C 5 -folic acid. To validate the performance of the DMHC 2D-LC/IDMS system on a complex sample, human plasma was analyzed for folic acid and the result was compared with that obtained using conventional single-column LC. The total run time of the DMHC 2D-LC system was 20 min, the same as that of the single-column LC system. The peak profile of the spiked 13 C 5 -folic acid obtained with single-column LC/MS was affected by matrix effects, but resolved with DMHC 2D-LC/MS, thus improving the accuracy of the analysis. The DMHC 2D-LC/IDMS system showed reliable performance in analyzing the target analyte in human plasma, eliminating matrix effects and saving analysis time., (© 2020 John Wiley & Sons, Ltd.)