Dr. Thibaud Aumond, Dr. Martin Esteves, Dr. Cristian Mocuta, Dr. Isabelle Batonneau-Gener, Dr. Julien Haines, Dr. Ricardo Faccio, Dr. Alexander Sachse
{"title":"In situ X-Ray Powder Diffraction Investigation on the Development of Zeolite-Templated Carbons in FAU Zeolite","authors":"Dr. Thibaud Aumond, Dr. Martin Esteves, Dr. Cristian Mocuta, Dr. Isabelle Batonneau-Gener, Dr. Julien Haines, Dr. Ricardo Faccio, Dr. Alexander Sachse","doi":"10.1002/cmtd.202400018","DOIUrl":"https://doi.org/10.1002/cmtd.202400018","url":null,"abstract":"<p>A time resolved <i>in situ</i> X-ray powder diffraction study using synchrotron radiation allowed for describing the evolution of the zeolite <b>FAU</b> structure during the development of a zeolite-templated carbon (ZTC) in its porous voids. During the ZTC formation the intensity decrease of most zeolite reflections and the simultaneous rise in intensity of the 222 reflection (of null intensity in the pristine zeolite) was observed. Full pattern profile fitting by Rietveld refinement allowed for achieving a detailed description of the underlying chemistry, with coincident pore filling with carbon atoms in specific positions and framework distortion. Monitoring the intensity profiles of the 222 reflection allowed assessment of the energetics of the ZTC formation. Our results contribute to a better understanding of the phenomena involved on the atomic scale in ZTC synthesis.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 12","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142861565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
MSc. Kristoffer Mega Herdlevær, MSc. Kasper Strandengen, Assoc. Prof. Dr. Camilla Løhre, Prof. Dr. Tanja Barth
{"title":"Determination of Sugar Concentrations in Aqueous Solution Using Multivariate Predictions Based on 1H-NMR Spectroscopy","authors":"MSc. Kristoffer Mega Herdlevær, MSc. Kasper Strandengen, Assoc. Prof. Dr. Camilla Løhre, Prof. Dr. Tanja Barth","doi":"10.1002/cmtd.202400004","DOIUrl":"https://doi.org/10.1002/cmtd.202400004","url":null,"abstract":"<p>Renewable chemicals from carbohydrate-rich wastes, like furfural and 5-hydroxymethylfurfural (HMF), are gaining prominence as alternatives to petroleum-based resources. Assessing the suitability of biomass as feedstock for furfural and HMF production requires knowledge of its composition. This study focuses on developing and validating predictive models for individual sugar concentrations in hydrolysates using quantitative 1H NMR data. Utilizing partial least square (PLS) regression, the dataset includes 137 NMR spectra of multi-component sugar standards (arabinose, fructose, galactose, glucose, mannose, maltose, sucrose, and xylose). The best-performing model achieved an R<sup>2</sup> of 0.987–0.999 and RMSECV of 0.37–1.56 mM and is based on the non-overlapping area of the NMR spectrum. Real-world samples were used for validation, resulting in predicted sugar concentrations with a mean standard deviation of 0.5 mM. This high accuracy and streamlined analysis process make these models practical for quantifying large sample sets, showcasing the reliability and accessibility of extracting statistical information from H-NMR data.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143115170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Felix Ott, Dr. Gudrun Gygli, Dr. Kersten S. Rabe, Prof. Dr. Christof M. Niemeyer
{"title":"Towards Precision Biocatalysis – Leveraging Inline NMR for Autonomous Experimentation in Flow Reactors","authors":"Felix Ott, Dr. Gudrun Gygli, Dr. Kersten S. Rabe, Prof. Dr. Christof M. Niemeyer","doi":"10.1002/cmtd.202400049","DOIUrl":"https://doi.org/10.1002/cmtd.202400049","url":null,"abstract":"<p>Reactor automation is a transformative force for chemical processes, but the potential of reaction monitoring for machine-assisted autonomous biocatalytic reaction optimization is still largely unexplored. To address this gap, we report on automated reactor optimization for biocatalytic flow-through microreactors. For this purpose, the inline NMR analysis of an enzymatically catalyzed stereoselective reduction of a prochiral diketone was combined with a self-developed open-source analysis and control software. The algorithm is continuously fed with spectra from a benchtop NMR instrument acquired from a reaction solution from a microreactor filled with biocatalytically active materials and adjusts the flow rate of the pumps to achieve predetermined target concentrations of the product. We show that through this automated coupling of data analysis and process parameterization, for example, maximum conversion efficiency can be achieved for a given bioreactor. This work illustrates the potential of inline NMR reaction monitoring for biocatalytic processes and provides a starting point for innovation to develop automated processes for precision biocatalysis through integrated data analysis.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 12","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142851338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Analysis of Macromolecular Size Distributions in Concentrated Solutions","authors":"Dr. Sumit K. Chaturvedi, Dr. Peter Schuck","doi":"10.1002/cmtd.202400035","DOIUrl":"https://doi.org/10.1002/cmtd.202400035","url":null,"abstract":"<p>The solution state of macromolecules in concentrated solutions impacts fields ranging from cell biology, to colloid chemistry and engineering of protein pharmaceuticals. Dependent on the interplay between repulsive and weakly attractive forces, proteins may exhibit oligomerization, aggregation, crystallization, liquid-liquid phase separation, or the formation of multiprotein complexes. The particle size-distribution is a key characteristic, but difficult to determine when interparticle distances are on the order of their size and macromolecular motion is coupled through hydrodynamic interactions. Here we extend sedimentation velocity analytical ultracentrifugation to measure macromolecular size distributions under these conditions: We apply results from statistical fluid mechanics for the concentration-dependence of hindered settling and diffusion, embedded in a mean-field approximation that can resolve coupled sedimentation and diffusion processes of different sized species given experimental sedimentation data. This is combined with a description of transient optical aberrations from lensing in the refractive index gradients associated with sedimentation boundaries (Wiener skewing). We demonstrate this approach in the application to protein solutions with macromolecular volume fractions up to ≈10 %, for example, resolving monomers and dimers of albumin at 140 mg/ml. This enables size-distribution analysis of proteins at concentrations of therapeutic antibody formations and close to physiological concentration in serum and cells.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 12","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400035","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142851468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jan Kulenkampff, Christian Armbruster, Johanna Drolshagen, Celine Regnat, Tina Wienold, Luisa Spari, Jana Fix, Tabea Sterbak, Dr. Harald Scherer, Prof. Dr. Ingo Krossing
{"title":"Cover Picture: Video Documented Upscaled Synthesis of Salts of the Parent Carbaborate Ion [CB11H12]−, its Undecafluorinated Form [CHB11F11]− and Useful Starting Materials for its Introduction (Chem. Methods 11/2024)","authors":"Jan Kulenkampff, Christian Armbruster, Johanna Drolshagen, Celine Regnat, Tina Wienold, Luisa Spari, Jana Fix, Tabea Sterbak, Dr. Harald Scherer, Prof. Dr. Ingo Krossing","doi":"10.1002/cmtd.202481101","DOIUrl":"https://doi.org/10.1002/cmtd.202481101","url":null,"abstract":"<p><b>The Front Cover</b> shows the molecular structure of the undecafluorinated carbaborate anion [CHB<sub>11</sub>F<sub>11</sub>]<sup>−</sup>, as well as its precursor [CB<sub>11</sub>H<sub>12</sub>]<sup>−</sup> and the starting material [BH<sub>4</sub>]<sup>−</sup>. Detailed synthetic protocols for the synthesis of [CB<sub>11</sub>H<sub>12</sub>]<sup>−</sup> and [CHB<sub>11</sub>F<sub>11</sub>]<sup>−</sup> in large scales are provided by Ingo Krossing and co-workers in their Research Article. To facilitate the reproduction of the preparation of these compounds, the synthesis has been filmed and instructive videos are provided with the publication. Starting from the [CHB<sub>11</sub>F<sub>11</sub>]<sup>−</sup> anion, the authors tested its chemical properties and synthesized its trityl ([Ph<sub>3</sub>C]<sup>+</sup>) as well as its silver salt ([Ag(<i>o</i>dfb)<sub>2</sub>]<sup>+</sup>) and used them for a representative hydrosilylation reaction, as well as the oxidation of ferrocene and the ‘magic blue’ amine N(4-C<sub>6</sub>H<sub>4</sub>Br)<sub>3</sub>. More information can be found in the Research Article by I. Krossing and co-workers (DOI: 10.1002/cmtd.202400011).\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 11","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202481101","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142665965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dr. Xuefeng Shen, Dr. Jiajia Ding, Dr. Yingchun Ye, Prof. Dr. Hongxing Liu, Prof. Dr. Zaiku Xie
{"title":"Ingrown Leading to Hierarchical SAPO-34 with High Catalytic Activity","authors":"Dr. Xuefeng Shen, Dr. Jiajia Ding, Dr. Yingchun Ye, Prof. Dr. Hongxing Liu, Prof. Dr. Zaiku Xie","doi":"10.1002/cmtd.202400027","DOIUrl":"https://doi.org/10.1002/cmtd.202400027","url":null,"abstract":"<p>Introduce meso/macro-pore to the crystal of SAPO-34 is an efficient way to overcome its inherent diffusion resistance. Porogen, formation hierarchical structure by inhibiting the growth of the crystals, would increase the costs of SAPO-34 and is usually not friendly to environment. In this work, an ingrowth route to synthesize hierarchical SAPO-34 is developed by secondary silica addition via hydrothermal method. Hierarchical porous SAPO-34 was synthesized without adding any pore-generating agent. Secondary silica addition put off the Si and P transferring from the mother liquid to gel phase, leading to the chemical composition of surface first meeting to the crystallization conditions. Then the crystallization started from the surface to the inside of the amorphous particles. The hierarchical SAPO-34 showed numerous meso/macro pore structure and weaker acid strength. So the hierarchical SAPO-34 showed better catalytic activity than the conventional SAPO-34 in MTO reaction.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 2","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400027","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143380485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dr. Konstantin S. Rodygin, Dr. Alexander S. Bogachenkov, Dr. Yulia V. Gyrdymova, Anton N. Potorochenko
{"title":"13C-Labeling as a Method in Organic Synthesis, Catalysis and Biochemical Applications","authors":"Dr. Konstantin S. Rodygin, Dr. Alexander S. Bogachenkov, Dr. Yulia V. Gyrdymova, Anton N. Potorochenko","doi":"10.1002/cmtd.202400045","DOIUrl":"https://doi.org/10.1002/cmtd.202400045","url":null,"abstract":"<p>The incorporation of a labeled <sup>13</sup>C atom provides a plethora of opportunities in organic synthesis and catalysis. Structure of complicated compounds can be successfully confirmed using the precise location of a labeled carbon atom; a reaction mechanism can be supported by the presence or absence of a label in reaction products; metabolic pathways of biological compounds can be found out depending on labeled products; the concentration of analyzed labeled compounds can be significantly lower for recording and successful interpretation of NMR spectra, <i>etc</i>. This review aims to provide a researcher with common and promising strategies for the synthesis of labeled compounds so that a researcher can select an appropriate route to save valuable labeled material. All the labeling approaches were considered based on starting labeling source. Ideological aim of the review is to demonstrate the opportunities for label incorporation so that a researcher can find the improvements in his own topic using labeled compounds. Utilization of labeled compounds in catalysis and studying reaction mechanisms were also considered to demonstrate the capabilities of labeled compounds, which may be useful in specific tasks and applications. The potential of labeled compounds in bioactive compound metabolite analysis was demonstrated on selected examples.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"5 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400045","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143114278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mayara da Silva Santos, Dr. Robert Medel, Simon Kruse, Max Flach, Olesya S. Ablyasova, Martin Timm, Prof. Dr. Bernd von Issendorff, Dr. Konstantin Hirsch, Dr. Vicente Zamudio-Bayer, Prof. Dr. Sebastian Riedel, Prof. Dr. J. Tobias Lau
{"title":"Direct Spectroscopic Confirmation of the Oxygen-Centered Diradical Character of the Tetraoxidorhenium(VII) Cation [Re(O)4]+","authors":"Mayara da Silva Santos, Dr. Robert Medel, Simon Kruse, Max Flach, Olesya S. Ablyasova, Martin Timm, Prof. Dr. Bernd von Issendorff, Dr. Konstantin Hirsch, Dr. Vicente Zamudio-Bayer, Prof. Dr. Sebastian Riedel, Prof. Dr. J. Tobias Lau","doi":"10.1002/cmtd.202400023","DOIUrl":"https://doi.org/10.1002/cmtd.202400023","url":null,"abstract":"<p>Mononuclear inorganic diradical species are scarce. Here, we confirm, via X-ray absorption spectroscopy in the gas phase combined with computational studies, the oxygen-centered diradical character of the tetraoxidorhenium(VII) cation. A dioxido-superoxido isomer, close in energy to the diradical, is also found, where rhenium appears in its rare oxidation state of +6. Addition of one or two hydrogen atoms to [Re,O<sub>4</sub>]<sup>+</sup> forms hydroxido ligands, and strongly disfavors isomers with any oxygen-oxygen bond. This adds spectroscopic characterization of the rhenium oxidation state and the nature of ligands to the known ability of [Re,O<sub>4</sub>]<sup>+</sup> to perform two consecutive hydrogen-atom abstraction reactions from methane, and demonstrates that pentaatomic [Re,O<sub>4</sub>]<sup>+</sup> combines a metal center in its highest oxidation state with two oxygen-centered radical ligands in a highly reactive species.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 12","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400023","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142851578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jan Kulenkampff, Christian Armbruster, Johanna Drolshagen, Celine Regnat, Tina Wienold, Luisa Spari, Jana Fix, Tabea Sterbak, Dr. Harald Scherer, Prof. Dr. Ingo Krossing
{"title":"Video Documented Upscaled Synthesis of Salts of the Parent Carbaborate Ion [CB11H12]−, its Undecafluorinated Form [CHB11F11]− and Useful Starting Materials for its Introduction","authors":"Jan Kulenkampff, Christian Armbruster, Johanna Drolshagen, Celine Regnat, Tina Wienold, Luisa Spari, Jana Fix, Tabea Sterbak, Dr. Harald Scherer, Prof. Dr. Ingo Krossing","doi":"10.1002/cmtd.202400011","DOIUrl":"https://doi.org/10.1002/cmtd.202400011","url":null,"abstract":"<p>In this work, we present our improved protocols for the single batch syntheses of approximately 32 g of [NHMe<sub>3</sub>][CB<sub>11</sub>H<sub>12</sub>] and 10 g of Na[CHB<sub>11</sub>F<sub>11</sub>] as well as salt metathesis reactions, granting access to useful starting materials to introduce the [CHB<sub>11</sub>F<sub>11</sub>]<sup>−</sup> anion. This includes the trityl cation [Ph<sub>3</sub>C]<sup>+</sup> and the bis-1,2-difluorobenzene-silver(I)-complex [Ag(<i>o</i>dfb)<sub>2</sub>]<sup>+</sup>, as well as some applications of the shown compounds. The described methodology allows the synthesis of large amounts of both the [CB<sub>11</sub>H<sub>12</sub>]<sup>−</sup> and the [CHB<sub>11</sub>F<sub>11</sub>]<sup>−</sup> anion and therefore making them accessible for further reactions. To facilitate the reproducibility, we present video tutorials of the synthetic steps towards Na[CHB<sub>11</sub>F<sub>11</sub>].</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 11","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400011","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142666104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rebecca M. Neeser, Prof. Bruno Correia, Prof. Philippe Schwaller
{"title":"FSscore: A Personalized Machine Learning-Based Synthetic Feasibility Score","authors":"Rebecca M. Neeser, Prof. Bruno Correia, Prof. Philippe Schwaller","doi":"10.1002/cmtd.202400024","DOIUrl":"https://doi.org/10.1002/cmtd.202400024","url":null,"abstract":"<p>Determining whether a molecule can be synthesized is crucial in chemistry and drug discovery, as it guides experimental prioritization and molecule ranking in <i>de novo</i> design tasks. Existing scoring approaches to assess synthetic feasibility struggle to extrapolate to new chemical spaces or fail to discriminate based on subtle differences such as chirality. This work addresses these limitations by introducing the Focused Synthesizability score (FSscore), which uses machine learning to rank structures based on their relative ease of synthesis. First, a baseline trained on an extensive set of reactant-product pairs is established, which is then refined with expert human feedback tailored to specific chemical spaces. This targeted fine-tuning improves performance on these chemical scopes, enabling more accurate differentiation between molecules that are hard and easy to synthesize. The FSscore showcases how a human-in-the-loop framework can be utilized to optimize the assessment of synthetic feasibility for various chemical applications.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"4 11","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202400024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142666060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}