bluuebunny/crossref_metadata_2025 huggingface dataset:
Systematic Dissection of an Aminopyrrolic Cage Receptor for β‐Glucopyranosides Reveals the Essentials for Effective Recognition
Authors: Francesconi Oscar , Gentili Matteo , Nativi Cristina , Ardá Ana , Cañada F. Javier , Jiménez‐Barbero Jesús , Roelens Stefano
Published: April 2014

A set of structures designed for the recognition of glucosides has been obtained by systematically destructuring a tripodal aminopyrrolic cage receptor that selectively recognizes octyl‐β‐D‐glucopyranoside (OctβGlc). NMR spectroscopy and isothermal titration calorimetry binding measurements showed that cleavage of one pillar of the cage was beneficial to the binding properties of the receptor, as long as two residual amino groups of the cleaved pillar were present. Removal of these two residual amino groups produced a dramatic loss of affinity for OctβGlc of the resulting monocyclic analogue of the parent cage receptor. A significant improvement in the binding ability was achieved by replacing one pillar with two aminopyrrolic hydrogen‐bonding arms, despite the loss of a preorganized structure. In contrast to the cage receptor, recognition of OctβGlc was observed, even in a competitive medium (30 % DMF in chloroform). Structural studies in solution, carried out through NMR spectroscopy and molecular modeling calculations, led to the elucidation of the 3D binding modes of the side‐armed monocyclic receptors; this highlighted the key role of the amino groups and demonstrated the occurrence of a rotaxane‐like complex, which featured the octyl chain of the glucoside threaded through the macrocyclic ring.
ChemInform Abstract: Structural and Spectroscopic Studies of Fluoroprotactinates.
Authors: De Sio Stephanie M. , Wilson Richard E.
Published: April 2014

The compounds (NH4)2PaF7 (I), K2PaF7 (II), Rb2PaF7 (III), Cs2PaF7 (IV), Na3PaF8 (V), (Me4N)2(H3O)PaF8 (VI), and (Me4N)PaF6 (VII) are synthesized from a 5 M aq.
ChemInform Abstract: Synthesis, Structure, and Luminescent Property of the Templated Borate [C9H14N]· [B5O6(OH)4].
Authors: Pan Chun‐Yang , Zhong Li‐Juan , Lu Jing , Li Da‐Guang , Zhao Feng‐Hua , Yang Hong‐Mei
Published: April 2014

[BupyH]·[B5O6(OH)4] is prepared under solvothermal conditions from H3BO3, N‐butyl‐pyridinium (Bupy) bromide, Al(O‐iPr)3, H2O, and pyridine, (autoclave, 180 °C, 7 d).
ChemInform Abstract: Filling in the Holes: Structural and Magnetic Properties of the Chemical Pressure Stabilized LnMnxGa3 (Ln: Ho—Tm; x < 0.15).
Authors: Fulfer Bradford W. , McAlpin Jacob D. , Engelkemier Joshua , McCandless Gregory T. , Prestigiacomo Joseph , Stadler Shane , Fredrickson Daniel C. , Chan Julia Y.
Published: April 2014

High‐quality single crystals of the title compounds are prepared from the constituent elements using Ga as a self‐flux (alumina crucible, 1150 °C, 5 h).
ChemInform Abstract: REBe2B5O11 (RE: Y, Gd): Rare‐Earth Beryllium Borates as Deep‐Ultraviolet Nonlinear‐Optical Materials.
Authors: Yan Xue , Luo Siyang , Lin Zheshuai , Yao Jiyong , He Ran , Yue Yinchao , Chen Chuangtian
Published: April 2014

Colorless block single crystals of the title compounds are obtained from a melt of Y2O3 or Gd2O3, BeO, B2O3, and Li2O (Pt crucible, 1100 °C).
ChemInform Abstract: Ba8Sn4S15: A Strong Second Harmonic Generation Sulfide with Zero‐Dimensional Crystal Structure.
Authors: Luo Zhong‐Zhen , Lin Chen‐Sheng , Zhang Wei‐Long , Zhang Hao , He Zhang‐Zhen , Cheng Wen‐Dan
Published: April 2014

The new title compound is synthesized by solid state reaction of Sn, BaS, and sulfur (graphite crucible, 1073 K, 48 h, <75% yield).
ChemInform Abstract: [Bi4]6‐ — The Zintl Anion with the Highest Charge per Atom Obtained from Solution.
Authors: Benda Christina B. , Faessler Thomas F.
Published: April 2014

Dark blue crystals of K6[Bi4](NH3)8 are obtained from a solution of the Zintl phase KBi in liquid NH3 in the presence of 18‐crown‐6 (‐70 °C, 3 months; 20‐30% yield).
ChemInform Abstract: Dicobalt‐μ‐oxo Polyoxometalate Compound, [(α2‐P2W17O61Co) 2O]14‐: A Potent Species for Water Oxidation, C—H Bond Activation, and Oxygen Transfer.
Authors: Barats‐Damatov Delina , Shimon Linda J. W. , Weiner Lev , Schreiber Roy E. , Jimenez‐Lozano Pablo , Poblet Josep M. , de Graaf Coen , Neumann Ronny
Published: April 2014

The highly reactive species [(α2‐P2W17O61Co) 2O]14‐ containing a nearly linear Co—O—Co moiety is prepared by oxidation of [α2‐P2W17O61Co II(H2O)]8‐ and characterized by single crystal XRD, 31 NMR, Raman, and UV/VIS spectroscopy, and DFT calculations.
ChemInform Abstract: Very Small (3—6 Atoms) Gold Cluster Catalyzed Carbon—Carbon and Carbon—Heteroatom Bond‐Forming Reactions in Solution.
Authors: Oliver‐Meseguer Judit , Leyva‐Perez Antonio , Corma Avelino
Published: April 2014

Several gold‐catalyzed reactions are investigated regarding the role of catalytically active Au—clusters generated in situ.
ChemInform Abstract: Efficient Transesterification Resolution of 2‐(4‐Chlorophenyl)‐4‐hydroxytetrahydropyran with Magnetically Separable Immobilized Lipase.
Authors: Xue Ping , Kang Zhen‐Zhen , Lai Xiao‐Yong , Qu Guan‐Qun , Li Yuan‐Yuan
Published: April 2014

Lipase immobilized on a core—shell MnFe2O4@SiO2— (CH2)3—NH2 carrier exhibits excellent catalytic activity for resolving racemic pyranol (I) via transesterification using vinyl acetate.
ChemInform Abstract: Asymmetric Hydrogenation of β‐Keto Sulfonamides and β‐Keto Sulfones with a Chiral Cationic Ruthenium Diamine Catalyst.
Authors: Huang Xiao‐Fei , Zhang Shao‐Yun , Geng Zhi‐Cong , Kwok Chun‐Yuen , Liu Peng , Li Hai‐Yan , Wang Xing‐Wang
Published: April 2014

The highly efficient asymmetric reduction of title compounds (I), (III), and (V) using a phosphine‐free chiral Ru‐complex furnishing corresponding β‐hydroxy sulfonamides and ‐sulfones (II), (IV), and (VI), resp., is described.
ChemInform Abstract: Decatungstate Photocatalyzed Benzylation of Alkenes with Alkylaromatics.
Authors: Qrareya Hisham , Ravelli Davide , Fagnoni Maurizio , Albini Angelo
Published: April 2014

The direct benzylation of electrophilic alkenes with alkylbenzenes is achieved by decatungstate photocatalysis in a high ionic strength medium.
ChemInform Abstract: Rapid and Ecofriendly Esterification of Alcohols with 2‐Acylpyridazinones.
Authors: Kim Bo Ram , Sung Gi Hyeon , Ryu Ki Eun , Kim Jeum‐Jong , Yoon Yong‐Jin
Published: April 2014

The new method for the esterification of various aromatic and aliphatic alcohols occurs under solvent‐free conditions within short reaction times and is also applicable to mercaptoethanol (VII) as well as benzeneselenol (X).
ChemInform Abstract: The Development of N‐Aryl Trifluoroacetimidate‐Based Benzyl and Allyl Protecting Group Reagents.
Authors: Tsabedze Sinele B. , Kabotso Daniel E. K. , Pohl Nicola L. B.
Published: April 2014

p‐Nitrophenyl substituted imidates (II) are efficient benzylating and allylating reagents in the presence of catalytic amounts of TmsOTf.
ChemInform Abstract: A Two‐Step Sequence to Ethyl α‐Fluorocyclopropanecarboxylates Through MIRC Reaction of Ethyl Dichloroacetate and Highly Regioselective Fluorination.
Authors: Zhang Min , Gong Yuefa , Wang Weizhou
Published: April 2014

The two‐step sequence yields monofluorinated cyclopropane derivatives which are of pharmaceutical interest due to their biological activities.
ChemInform Abstract: Dihaloiodoarenes: α,α‐Dihalogenation of Phenylacetate Derivatives.
Authors: Tao Jason , Tran Richard , Murphy Graham K.
Published: April 2014

Diazoacetate derivatives react with Ph‐ICl2 or Tol‐IF2 to afford gem‐dichlorinated or gem‐difluorinated products in the presence of a Lewis base or Lewis acid, respectively.
ChemInform Abstract: An Alternate Preparation of 3,4,5‐Trimethoxyphenol.
Authors: Qin Qi‐xue , Yang Jian , Chen Duo‐zhi , Yang Bo , Zhang Ji
Published: April 2014

All products are purified by recrystallization.
ChemInform Abstract: Asymmetric Reduction of 2,2‐Dimethyl‐6‐(2‐oxoalkyl/oxoaryl)‐1,3‐dioxin‐4‐ones and Application to the Synthesis of (+)‐Yashabushitriol.
Authors: Fang Zhijia Amphi , Clarkson Guy J. , Wills Martin
Published: April 2014

The asymmetric transfer hydrogenation of various ketones (I) is achieved in high enantiomeric excess using a chiral ruthenium complex catalyst.
ChemInform Abstract: Amino Acid Intercalated Layered Double Hydroxide Catalyzed Chemoselective Methylation of Phenols and Thiophenols with Dimethyl Carbonate.
Authors: Subramanian Thirumeni , Dhakshinamoorthy Amarajothi , Pitchumani Kasi
Published: April 2014

Sixteen amino acids are intercalated into Mg‐Al layered double hydroxides by the reconstruction method and utilized as catalysts in the title reaction.
ChemInform Abstract: Tailoring the S‐Selectivity of 2‐Succinyl‐5‐enolpyruvyl‐6‐hydroxy‐3‐cyclohexene‐1‐carboxylate Synthase (MenD) from Escherichia coli.
Authors: Pohl Martina , et al. et al.
Published: April 2014

S‐selective EcMenD variants of the thiamine diphosphate (TDP)‐dependent enzyme 2‐succinyl‐5‐enolpyruvyl‐6‐hydroxy‐3‐cyclohexene‐1‐carboxylate synthase from Escherichia coli (E.C.
ChemInform Abstract: A One‐Pot Asymmetric Synthesis of Homoallylic Amines via Indium‐Mediated Allylation of Hydrazones.
Authors: Lee Jun Hyuk , Nam Tae Kyu , Satyender Apuri , Jang Doo Ok
Published: April 2014

Optimum reaction conditions are examined for the asymmetric addition of allyl iodide (III) to hydrazones which are prepared in situ from (I) and aldehydes (II).
ChemInform Abstract: Highly Diastereoselective Synthesis of Dispiro[1,4‐dithiane/dithiepane]bisoxindoles via Stevens Rearrangement.
Authors: Muthusamy Sengodagounder , Selvaraj Karuppu
Published: April 2014

The rhodium‐catalyzed reaction of oxindoles (I) and (IV) with diazooxindoles involves the formation of a sulfonium ylide intermediate.
ChemInform Abstract: Polyethylene Glycol (PEG‐400) as an Efficient and Recyclable Reaction Medium for the Synthesis of 2‐Aroylbenzofurans.
Authors: Zhao Sanhu , Wang Xiaoju , Zhang Liwei
Published: April 2014

The reaction is conducted under mild conditions and uses an inexpensive and recyclable reaction medium.
ChemInform Abstract: Broensted Acid Catalyzed Enantioselective Indole Aza‐Claisen Rearrangement Mediated by an Arene CH—O Interaction.
Authors: Maity Pradip , Pemberton Ryan P. , Tantillo Dean J. , Tambar Uttam K.
Published: April 2014

The reaction tolerates a wide range of aromatic and heteroaromatic substituents at the allylic double bond and affords the products with high yields and enantioselectivities [cf.
ChemInform Abstract: Batch and Flow Photochemical Benzannulations Based on the Reaction of Ynamides and Diazo Ketones. Application to the Synthesis of Polycyclic Aromatic and Heteroaromatic Compounds.
Authors: Willumstad Thomas P. , Haze Olesya , Mak Xiao Yin , Lam Tin Yiu , Wang Yu‐Pu , Danheiser Rick L.
Published: April 2014

A two‐stage tandem benzannulation/cyclization strategy for the synthesis of the title compounds is presented.
ChemInform Abstract: Molecular Iodine‐Mediated Cascade Reaction of 2‐Alkynylbenzaldehyde and Indole: An Easy Access to Tetracyclic Indoloazulene Derivatives.
Authors: Gawande Sachin D. , Kavala Veerababurao , Zanwar Manoj R. , Kuo Chun‐Wei , Huang Hsiu‐Ni , He Chiu‐Hui , Kuo Ting‐Shen , Yao Ching‐Fa
Published: April 2014

The synthesis of iodo‐substituted tetracyclic indole fused azulene derivatives is achieved by reaction of various phenylethynyl benzaldehydes and different indoles in the presence of molecular iodine.
ChemInform Abstract: A Three‐Way Switchable Process for Suzuki Cross‐Coupling, Hydrodehalogenation, or an Assisted Tandem Hydrodehalogenation and Suzuki Cross‐Coupling Sequence.
Authors: Sandtorv Alexander H. , Bjoersvik Hans‐Rene
Published: April 2014

The Suzuki reaction of 4,5‐iodoimidazole is thoroughly investigated.
ChemInform Abstract: Asymmetric Synthesis and Effect of Absolute Stereochemistry of YCZ‐2013, a Brassinosteroid Biosynthesis Inhibitor.
Authors: Oh Keimei , Yamada Kazuhiro , Yoshizawa Yuko
Published: April 2014

The four stereoisomers of YCZ‐2013 are synthesized and their effects on Arabidopses stem elongation is studied.
ChemInform Abstract: Enantioselective Synthesis of Coumarin Derivatives Catalyzed by Primary Amines.
Authors: Lee Yen‐Te , Das Utpal , Chen Yi‐Ru , Lee Chia‐Jui , Chen Chi‐Han , Yang Mei‐Chun , Lin Wenwei
Published: April 2014

Michael addition reaction of ketones with coumarin derivatives in the presence of cinchona type catalyst provides optically active coumarins with excellent enantioselectivity.
ChemInform Abstract: Novel One‐Pot Synthesis of Xanthones via Sequential Fluoride Ion‐Promoted Fries‐Type Rearrangement and Nucleophilic Aromatic Substitution.
Authors: Fujimoto Yuuki , Itakura Ryohei , Hoshi Hiroki , Yanai Hikaru , Ando Yoshio , Suzuki Keisuke , Matsumoto Takashi
Published: April 2014

The title method allows the efficient conversion of 2‐fluorobenzoate derivatives into xanthones that have significant steric congestion around the C9 carbonyl and which are not readily available by conventional methods.
ChemInform Abstract: Copper(I) Iodide/Boron Trifluoride Etherate‐Cocatalyzed Aerobic Dehydrogenative Reactions Applied in the Synthesis of Substituted Heteroaromatic Imidazo[1,2‐a]pyridines.
Authors: Cai Zhong‐Jian , Wang Shun‐Yi , Ji Shun‐Jun
Published: April 2014

The oxidative cyclocondensation of 2‐aminopyridines with aryl alkyl ketones is investigated to provide a synthesis of imidazo[1,2‐a]pyridines.
ChemInform Abstract: Synthesis and Structure of Condensed Triazolo‐ and Tetrazolopyrimidines.
Authors: Sirakanyan Samvel N. , Geronikaki Athina , Spinelli Domenico , Hovakimyan Anush A. , Noravyan Azat S.
Published: April 2014

Hydrazino derivatives (I) and (VI) undergo ring closure reactions by treatment with triethyl orthoformate (II), formic acid (IV) and nitrous acid to give the title compounds in good yields.
ChemInform Abstract: Synthesis of Conformationally Restricted C2 Symmetric Macrodiolides via Head to Tail Dimerization of Carbonyl Ylides.
Authors: Muthusamy Sengodagounder , Sivaguru Manickasamy
Published: April 2014

A simple procedure is developed for the synthesis of the title species via rhodium(II) carbenoid induced head to tail dimerization of carbonyl ylides having unactivated alkyne or alkene functionalities.
ChemInform Abstract: Synthesis and Suzuki—Miyaura Cross‐Coupling of Enantioenriched Secondary Potassium β‐Trifluoroboratoamides: Catalytic, Asymmetric Conjugate Addition of Bisboronic Acid and Tetrakis(dimethylamino)diboron to α,β‐Unsaturated Carbonyl Compounds.
Authors: Molander Gary A. , Wisniewski Steven R. , Hosseini‐Sarvari Mona
Published: April 2014

Enantioenriched title compounds are synthesized via a new asymmetric, copper‐catalyzed addition of tetrahydroxydiboron to various α,β‐unsaturated amides, ketones, and esters in high yields and enantioselectivities.
ChemInform Abstract: Gold‐Catalyzed Hydroarylation of Aryl Alkynylphosphonates for the Synthesis of Phosphacoumarins.
Authors: Kim Cheol‐Eui , Ryu Taekyu , Kim Sanghyuck , Lee Kooyeon , Lee Chang‐Hee , Lee Phil Ho
Published: April 2014

An intramolecular hydroarylation of aryl alkynylphosphonates (I) and (III) promoted by a gold/silver catalyst affords phosphacoumarins (II) and (IV), resp., in moderate to good yields via a 6‐endo‐dig cyclization.
ChemInform Abstract: Total Synthesis of (+)‐Asperolide C by Iridium‐Catalyzed Enantioselective Polyene Cyclization.
Authors: Jeker Oliver F. , Kravina Alberto G. , Carreira Erick M.
Published: April 2014

The first total synthesis of the target compound is described.
ChemInform Abstract: Three New Iridoid Glycosides from the Fruit of Gardenia jasminoides var. radicans
Authors: Qin Fang‐min , Meng Ling‐jie , Zou Hui‐liang , Zhou Guang‐xiong
Published: April 2014

isolation and structure elucidation
ChemInform Abstract: Exploring Nanoscaled Matter from Speciation to Phase Diagrams: Metal Phosphide Nanoparticles as a Case of Study
Authors: Carenco Sophie , Portehault David , Boissiere Cedric , Mezailles Nicolas , Sanchez Clement
Published: April 2014

Review: [128 refs.
ChemInform Abstract: Carbon Nanofluidics of Rapid Water Transport for Energy Applications
Authors: Park Hyung Gyu , Jung Yousung
Published: April 2014

Review: 55 refs.
ChemInform Abstract: Transition Metal Complexes Supported by Highly Basic Imidazolin‐2‐iminato and Imidazolin‐2‐imine N‐Donor Ligands
Authors: Wu Xian , Tamm Matthias
Published: April 2014

Review: 186 refs.
ChemInform Abstract: Radical‐Mediated Brominations at Ring‐Positions of Carbohydrates ‐ 35 Years Later
Authors: Somsak Laszlo , Czifrak Katalin
Published: April 2014

Review: 130 refs.
ChemInform Abstract: Asymmetric Catalytic Anhydride Openings via Carbon‐Based Nucleophiles
Authors: Zheng Chen , Chen Fen‐Er
Published: April 2014

Review: 33 refs.
ChemInform Abstract: Manipulation of Micro‐ and Nanostructure Motion with Magnetic Fields
Authors: Rikken Roger S. M. , Nolte Roeland J. M. , Maan Jan C. , van Hest Jan C. M. , Wilson Daniela A. , Christianen Peter C. M.
Published: April 2014

Review: 117 refs.
ChemInform Abstract: Recent Progress in the Identification of BRAF Inhibitors as Anticancer Agents
Authors: El‐Nassan Hala Bakr
Published: April 2014

Review: 144 refs.
ChemInform Abstract: Updated Assessment of Possibilities and Limits for Solar Cells
Authors: Nayak Pabitra K. , Cahen David
Published: April 2014

Review: 53 refs.
ChemInform Abstract: Spectroscopy of Single Nanocrystals
Authors: Fernee Mark J. , Tamarat Philippe , Lounis Brahim
Published: April 2014

Review: 208 refs.
Thymochinon – das Gelbe vom Öl
Authors: Spiller Sandro , Mettke Peter , Siehl Hans‐Ullrich , Zeller Klaus‐Peter , Sicker Dieter , Berger Stefan
Published: April 2014

Thymochinon wurde durch Wasserdampfdestillation aus gelbem Schwarzkümmelöl gewonnen und durch Säulenchromatographie gereinigt. Alle analytischen Spektren sind vollständig entweder im Hauptteil oder in der supporting information wiedergegeben. Die NMR‐ sowie die Massenspektren werden eingehend interpretiert und mit theoretischen Berechnungen der 13C‐chemischen Verschiebungen verglichen. Das Projekt stellt eine Fortsetzung des Buches “Classics in Spectroscopy” von S. Berger und D. Sicker (Wiley‐VCH 2009) dar.
Lack of Efficacy of Drug Therapy in Preventing Takotsubo Cardiomyopathy Recurrence: A Meta‐analysis
Authors: Santoro Francesco , Ieva Riccardo , Musaico Francesco , Ferraretti Armando , Triggiani Giuseppe , Tarantino Nicola , Biase Matteo Di , Brunetti Natale Daniele
Published: April 2014

Background

Efficacy of chronic drug therapy in prevention of stress‐induced cardiomyopathy recurrences is not well established. We therefore aimed to evaluate in this meta‐analysis whether pharmacological treatment can effectively prevent takotsubo cardiomyopathy (TTC) recurrences, according to available studies.

Hypothesis

There is no evidence for preventing TTC recurrence by drug therapy.

Methods

After a PubMed search, we conducted a meta‐analysis of available studies (clinical nonrandomized registries) on efficacy of drug therapy in preventing recurrence of TTC.

Results

A total of 23 (4.5%) TTC recurrences occurred in the 511 patients included in the analysis. Seven studies on the effects of β‐blockers on prevention of TTC recurrence were evaluated; the odds ratio (OR) was 0.44 and the 95% confidence interval (CI) was 0.15‐1.31. In 5 studies on the effects of angiotensin‐converting enzyme inhibitors/angiotensin receptor blockers, the OR was 0.42 and the 95% CI was 0.08‐2.36; in 3 studies on statins, the OR was 0.74 and the 95% CI was 0.07‐7.3; and in 4 studies on aspirin, the OR was 0.33 with a 95% CI of 0.05‐2.17 (P value not significant in all cases).

Conclusions

A meta‐analysis of the efficacy of different medications through the clinical TTC registries available showed no clinical evidence for a standard drug treatment in the chronic management of TTC. β‐Blockers, angiotensin‐converting enzyme inhibitors/angiotensin receptor blockers, statins, and aspirin do not seem to significantly reduce recurrences of TTC. Randomized, adequately powered studies are needed to further assess this issue.
A comprehensive review on controls in molecular imaging: lessons from MMP‐2 imaging
Authors: Lebel Réjean , Lepage Martin
Published: April 2014

Metalloproteinases (MMPs), including MMP‐2, play critical roles in tissue remodeling and are involved in a large array of pathologies, including cancer, arthritis and atherosclerosis. Their prognostic value warranted a large investment or resources in the development of noninvasive detection methods, based on probes for many current clinical and pre‐clinical imaging modalities. However, the potential of imaging techniques is only matched by the complexity of the data they generate. This complexity must be properly assessed and accounted for in the early steps of probe design and testing in order to accurately determine the efficacy and efficiency of an imaging strategy. This review proposes basic rules for the evaluation of novel probes by addressing the specific case of MMP targeted probes. Copyright © 2014 John Wiley & Sons, Ltd.
Generation and characterization of nanobodies targeting PSMA for molecular imaging of prostate cancer
Authors: Evazalipour Mehdi , D'Huyvetter Matthias , Tehrani Bahram Soltani , Abolhassani Mohsen , Omidfar Kobra , Abdoli Shahriyar , Arezumand Roghaye , Morovvati Hamid , Lahoutte Tony , Muyldermans Serge , Devoogdt Nick
Published: April 2014

Nanobodies show attractive characteristics for tumor targeting in cancer diagnosis and therapy. A radiolabeled nanobody binding the prostate‐specific membrane antigen (PSMA) could offer a noninvasive strategy to select prostate cancer patients eligible for PSMA‐targeted therapies. We here describe the generation, production and in vivo evaluation of anti‐PSMA nanobodies. Nanobodies were derived from heavy‐chain‐only antibodies, raised in immunized dromedaries. Binding characteristics were evaluated through ELISA and flow cytometry. Selected nanobodies were radiolabeled with 99mTc at their hexahistidine tail, after which cell binding capacity and internalization were evaluated on PSMApos LNCaP and PSMAneg PC3 cell lines. In vivo tumor targeting was analyzed in both LNCaP and PC3 xenografted mice through SPECT/microCT and tissue sampling. A panel of 72 generated clones scored positive on ELISA, all contributing to three nanobody groups, of which group 3 dominated with 70 clones. ELISA and FACS analysis led to the selection of two dominant nanobodies. 99mTc‐labeled PSMA6 and PSMA30 both showed specific binding on LNCAP cells, but not on PC3 cells. 99mTc‐PSMA30 internalized significantly more in LNCaP cells compared to 99mTc‐PSMA6. Higher absolute tumor uptake and tumor‐to‐normal organ ratios were observed for 99mTc‐PSMA30 compared with 99mTc‐PSMA6 and a 99mTc‐control nanobody in LNCaP but not in PC3 tumor‐bearing mice. PSMA30 nanobody has improved targeting characteristics both in vitro as well as in vivo compared with PSMA6 and the control nanobody, and was therefore selected as our in‐house‐developed lead compound for PSMA targeting. Copyright © 2014 John Wiley & Sons, Ltd.
Monitoring hypoxia and vasculature during bevacizumab treatment in a murine colorectal cancer model
Authors: Heijmen L. , ter Voert E. G. W. , Punt C. J. A. , Heerschap A. , Oyen W. J. G. , Bussink J. , Sweep C. G. J. , Laverman P. , Span P. N. , de Geus‐Oei L. F. , Boerman O. C. , van Laarhoven H. W. M.
Published: April 2014

The purpose of this study was to assess the effect of bevacizumab on vasculature and hypoxia in a colorectal tumor model. Nude mice with subcutaneous LS174T tumors were treated with bevacizumab or saline. To assess tumor properties, separate groups of mice were imaged using 18F‐Fluoromisonidazole (FMISO) and 18F‐Fluorodeoxyglucose (FDG) positron emission tomography or magnetic resonance imaging before and 2, 6 and 10 days after the start of treatment. Tumors were harvested after imaging to determine hypoxia and vascular density immunohistochemically. The T2* time increased significantly less in the bevacizumab group. FMISO uptake increased more over time in the control group. Vessel density significantly decreased in the bevacizumab‐treated group. The Carbonic anhydrase 9 (CAIX) and glucose uptake transporter 1 (GLUT1) fractions were higher in bevacizumab‐treated tumors. However, the hypoxic fraction showed no significant difference. Bevacizumab led to shorter T2* times and higher GLUT1 and CAIX expression, suggesting an increase in hypoxia and a higher glycolytic rate. This could be a mechanism of resistance to bevacizumab. The increase in hypoxia, however, could not be demonstrated by pimonidazole/FMISO, possibly because distribution of these tracers is hampered by bevacizumab‐induced effects on vascular permeability and perfusion. Copyright © 2014 John Wiley & Sons, Ltd.
The reciprocal linear QUEST analysis method facilitates the measurements of chemical exchange rates with CEST MRI
Authors: Randtke Edward A. , Chen Liu Qi , Pagel Mark D.
Published: April 2014

Magnetic resonance imaging (MRI) contrast media that are detected via chemical exchange saturation transfer (CEST) often require an accurate estimation of their chemical exchange rate, kex. A variety of analysis methods have been proposed to estimate kex, including the nonlinear QUEST analysis method that evaluates the CEST amplitude as a function of saturation time. We have derived a linear version of QUEST, termed the Reciprocal Linear QUEST (RL‐QUEST) method. Our simulations and experimental results show that RL‐QUEST performs as well as QUEST, while providing a more simplistic fitting procedure. Although CEST results should be acquired with saturation power that has a nutation rate that is faster than kex of the CEST agent, an exact determination of the saturation power is not required to accurately estimate kex with RL‐QUEST. This new analysis method requires a determination of the CEST agent's concentration, which is straightforward for the analysis of CEST agents in chemical solutions, but may be a limitation during in vivo CEST MRI studies. Based on the results of this study and previous studies, we provide recommendations for the linear analysis method that should be employed for each type of CEST MRI study. Copyright © 2014 John Wiley & Sons, Ltd.
Assessment of mutational profile of Japanese lung adenocarcinoma patients by multitarget assays: A prospective, single‐institute study
Authors: Serizawa Masakuni , Koh Yasuhiro , Kenmotsu Hirotsugu , Isaka Mitsuhiro , Murakami Haruyasu , Akamatsu Hiroaki , Mori Keita , Abe Masato , Hayashi Isamu , Taira Tetsuhiko , Maniwa Tomohiro , Takahashi Toshiaki , Endo Masahiro , Nakajima Takashi , Ohde Yasuhisa , Yamamoto Nobuyuki
Published: April 2014

BACKGROUND

Integration of mutational profiling to identify driver genetic alterations in a clinical setting is necessary to facilitate personalized lung cancer medicine. A tumor genotyping panel was developed and the Shizuoka Lung Cancer Mutation Study was initiated as a prospective tumor genotyping study. This study reports the frequency of driver genetic alterations in Japanese lung adenocarcinoma patients, and clinicopathologic correlations with each genotype.

METHODS

Between July 2011 and January 2013, 411 lung adenocarcinoma patients admitted to the Shizuoka Cancer Center were included in this study with their written informed consent. Surgically resected tissues, tumor biopsies, and/or body cavity fluids were collected and tested for 23 hotspot sites of driver mutations in 9 genes (EGFR, KRAS, BRAF, PIK3CA, NRAS, MEK1, AKT1, PTEN, and HER2), gene amplifications in 5 genes (EGFR, MET, PIK3CA, FGFR1, and FGFR2), and ALK, ROS1, and RET fusions.

RESULTS

Genetic alterations were detected in 54.3% (223 of 411) of all patients. The most common genetic alterations detected in this study were EGFR mutations (35.0%) followed by KRAS mutations (8.5%) and ALK fusions (5.0%). Concurrent genetic alterations were detected in 22 patients (5.4%), and EGFR mutations were observed in 16 patients as the most common partner for concurrent genetic alteration. Significantly more concurrent genetic alterations were observed in older patients.

CONCLUSIONS

This is one of the largest reports of a prospective tumor genotyping study on Japanese patients with adenocarcinoma. These data suggest that mutational profiling data using a multimutational testing platform would be valuable for expanding the range of molecular‐targeted therapeutics in lung cancer. Cancer 2014;120:1471–1481. © 2014 American Cancer Society.
RET gene mutations (genotype and phenotype) of multiple endocrine neoplasia type 2 and familial medullary thyroid carcinoma
Authors: Krampitz Geoffrey W. , Norton Jeffrey A.
Published: April 2014

The rapid technical advances in molecular biology and accelerating improvements in genomic and proteomic diagnostics have led to increasingly personalized strategies for cancer therapy. Such an approach integrates the genomic, proteomic, and molecular information unique to the individual to provide an accurate genetic diagnosis, molecular risk assessment, informed family counseling, therapeutic profiling, and early preventative management that best fits the particular needs of each patient. The discovery of mutations in the RET proto‐oncogene resulting in variable onset and severity of multiple endocrine neoplasia type 2 (MEN2) was the first step in developing direct genetic testing for at‐risk individuals. Patients with germline RET mutations may undergo risk assessment and appropriate intervention based on specific mutations. Moreover, family members of affected individuals receive counseling based on understanding of the genetic transmission of the disease. Increasingly, clinicians are able to make therapeutic choices guided by an informative biomarker code. Improvements in detection and management of patients with MEN2 resulting from understanding of the RET proto‐oncogene are evidence of the benefits of personalized cancer medicine. This review describes the discovery of the RET proto‐oncogene, the association between genotype and phenotype, and the role of mutation analysis on diagnosis and treatment of MEN2. Cancer 2014;120:1920–1931. © 2014 American Cancer Society.
Laterally Projecting Cerebrospinal Fluid‐Contacting Cells in the Lamprey Spinal Cord Are of Two Distinct Types
Authors: Jalalvand Elham , Robertson Brita , Wallén Peter , Hill Russell H. , Grillner Sten
Published: April 2014

Cerebrospinal fluid‐contacting (CSF‐c) cells are found in all vertebrates, but their function remains elusive. In the lamprey spinal cord, they surround the central canal and some have processes passing the gray matter to the lateral edge of the flattened spinal cord. Stimulation of CSF‐c cells at the central canal elicits GABAergic inhibitory postsynaptic potentials (IPSPs) in intraspinal stretch receptor neurons (edge cells). Here, we characterize laterally projecting CSF‐c cells according to their morphology, phenotype, and neuronal properties by using immunohistochemistry, retrograde tracing, calcium imaging, and whole‐cell recordings. We identify two types of CSF‐c cells. Type 1 cells have a bulb‐like ending that protrudes into the central canal and a lateral process that ramifies ventrolaterally and laterally with a dense plexus surrounding the mechanosensitive dendrites of the edge cells. Most type 1 cells fire spontaneous action potentials that are abolished by tetrodotoxin, and all display spontaneous excitatory postsynaptic potentials and IPSPs that remain in the presence of tetrodotoxin. GABA and somatostatin are colocalized in type 1 cells, and they express both GABA and glutamate receptors. Type 2 cells, on the other hand, have a flat ending protruding into the central canal and a laterally projecting process that ramifies only at the lateral edge. These cells show immunoreactivity to taurine, but they do not express GABA or somatostatin, nor do they have any active neuronal properties. Type 2 cells might be a form of glia. Type 1 CSF‐c cells are neurons and may play a modulatory role by influencing edge cells and thus the locomotor‐related sensory feedback. J. Comp. Neurol. 522:1753–1768, 2014. © 2014 Wiley Periodicals, Inc.
RF and microwave high‐Q floating active inductor design and implementation
Authors: Branchi P. , Pantoli L. , Stornelli V. , Leuzzi G.
Published: April 2014

In this paper, a high‐Q floating active inductor (FAI), suitable for RF and microwave applications, is presented. The proposed FAI is based on two cascaded pairs of highly linear capacitance gyrators, which provide a symmetric and reciprocal structure. The proposed FAI shows fully symmetrical two‐port characteristics, high quality factor and high linearity. As a feasibility demonstration, a prototype of the designed FAI has been fabricated, together with an LC series band‐pass filter. At the operating frequency, the real part of the impedance of the equivalent FAI is very low (Req = 0.0039 Ω), providing a very high quality factor. The filter has a central frequency of 430 MHz and a −3 dB bandwidth of about 9 MHz. Copyright © 2014 John Wiley & Sons, Ltd.
Fine needle aspiration cytology of a myoepithelioma presenting as a thyroid nodule
Authors: Narick Christina , Velosa Claudia , Pollice Philip , Silverman Jan
Published: April 2014

Myoepitheliomas are rare neoplasms that are typically found in the major and minor salivary glands and represent approximately 1.5% of all salivary gland neoplasms. We present a patient with an exophytic anterior midline neck mass, which was initially believed to be a thyroid isthmus nodule that underwent fine needle aspiration (FNA) biopsy. FNA cytologic evaluation reveals numerous plump spindle cells and a myxoid background, thus raising the possibility of rare benign mixed tumor of the thyroid. However, the resected specimen consists of predominately spindle cells with a minor component of chondromyxoid matrix, and no ductal epithelial cells, favoring a diagnosis of myoepithelioma. Although this lesion clinically and radiologically appeared to arise from the thyroid gland, at the time of resection, it was found to be adjacent to the thyroid isthmus and was ultimately diagnosed as a soft tissue myoepithelioma of the midneck. Diagn. Cytopathol. 2015;43:153–157. © 2014 Wiley Periodicals, Inc.
The MAP1B case: An old MAP that is new again
Authors: Villarroel‐Campos David , Gonzalez‐Billault Christian
Published: April 2014

The functions of microtubule‐associated protein 1B (MAP1B) have historically been linked to the development of the nervous system, based on its very early expression in neurons and glial cells. Moreover, mice in which MAP1B is genetically inactivated have been used extensively to show its role in axonal elongation, neuronal migration, and axonal guidance. In the last few years, it has become apparent that MAP1B has other cellular and molecular functions that are not related to its microtubule‐stabilizing properties in the embryonic and adult brain. In this review, we present a systematic review of the canonical and novel functions of MAP1B and propose that, in addition to regulating the polymerization of microtubule and actin microfilaments, MAP1B also acts as a signaling protein involved in normal physiology and pathological conditions in the nervous system. © 2014 Wiley Periodicals, Inc. Develop Neurobiol 74: 953–971, 2014
TGF‐β‐producing regulatory B cells induce regulatory T cells and promote transplantation tolerance
Authors: Lee Kang Mi , Stott Ryan T , Zhao Gaoping , SooHoo Julie , Xiong Wei , Lian Moh Moh , Fitzgerald Lindsey , Shi Shuai , Akrawi Elsie , Lei Ji , Deng Shaoping , Yeh Heidi , Markmann James F , Kim James I
Published: April 2014

Regulatory B (Breg) cells have been shown to play a critical role in immune homeostasis and in autoimmunity models. We have recently demonstrated that combined anti‐T cell immunoglobulin domain and mucin domain‐1 and anti‐CD45RB antibody treatment results in tolerance to full MHC‐mismatched islet allografts in mice by generating Breg cells that are necessary for tolerance. Breg cells are antigen‐specific and are capable of transferring tolerance to untreated, transplanted animals. Here, we demonstrate that adoptively transferred Breg cells require the presence of regulatory T (Treg) cells to establish tolerance, and that adoptive transfer of Breg cells increases the number of Treg cells. Interaction with Breg cells in vivo induces significantly more Foxp3 expression in CD4+CD25− T cells than with naive B cells. We also show that Breg cells express the TGF‐β associated latency‐associated peptide and that Breg‐cell mediated graft prolongation post‐adoptive transfer is abrogated by neutralization of TGF‐β activity. Breg cells, like Treg cells, demonstrate preferential expression of both C‐C chemokine receptor 6 and CXCR3. Collectively, these findings suggest that in this model of antibody‐induced transplantation tolerance, Breg cells promote graft survival by promoting Treg‐cell development, possibly via TGF‐β production.
Construction of Metal–Organic Frameworks Consisting of Dinuclear Metal Units Based on 5‐Hydroxyisophthalate and Flexible Dipyridyl Ligands
Authors: Li Xiaoju , Guan Tena , Guo Xiaofang , Li Xinxiong , Yu Zhenjiang
Published: April 2014

Four CoII and CuII complexes, namely, [Co2(ip‐OH)2(dpe)]n [1; ip‐OH = 5‐hydroxyisophthalate, dpe = 1,2‐di(4‐pyridyl)ethane], [Co2(ip‐OH)2(dpy)2]n(dpy)n(H2O)2n [2; dpy = 1,2‐di(4‐pyridyl)ethylene], [Cu2(ip‐OH)2(dpe)]n (3), and [Cu2(ip‐OH)2(dpy)]n (4), were hydrothermally synthesized and characterized. Single‐crystal X‐ray diffraction studies reveal that 1–4 consist of different dinuclear metal units, ip‐OH possesses different coordination modes, and both dpe and dpy adopt anti conformations. In 1, the ip‐OH ligands link dinuclear CoII units into a charge‐neutral [Co2(ip‐OH)2]n layer, which is further connected by pillars of dpe to generate a twofold interpenetrating 3D network. However, dinuclear CoII units in 2 are bridged by ip‐OH to form a 1D [Co2(ip‐OH)2]n chain, which is further connected by dpy into a 2D network consisting of the prismatic building blocks [Co8(ip‐OH)8(dpy)8]. Similarly to 1, 3 and 4 are twofold interpenetrating α‐Po architectures, despite the different ip‐OH bridging modes and different coordination geometries of the metal ions. The dinuclear units in 1–4 are well separated by ip‐OH and dpe/dpy ligands. The magnetic data of 1–3 are fitted according to the dimeric modes, and the results of the best fit indicate the existence of antiferromagnetic exchange between the metal centers in the dinuclear metal units.
Comparison of Electrochemical Oxidation of Flavonols and Calculated Proton Affinity and Electron Transfer Enthalpy in Water
Authors: Kummer Sina , Ruth Wolfgang , Kühn Oliver , Kragl Udo
Published: April 2014

Flavonols, a class of flavonoids, are present in flowers, fruits and vegetables. They are jointly responsible for antioxidant activity as free radical acceptors. The redox behaviour of myricetin, quercetin, isorhamnetin, fisetin, morin and kaempferol is investigated using cyclic and differential pulse voltammetry. Quantum chemical calculations of proton affinities and electron transfer enthalpies were performed to identify possible reactive sites and radical species to compare them with measured oxidation potentials of the flavonols. Regarding to their chemical structure, these flavonols showed an oxidation order: myricetin > quercetin > isorhamnetin > fisetin > morin > kaempferol.
A Novel Electrochemical Sensor for β2‐Agonists with High Sensitivity and Selectivity Based on Surface Molecularly Imprinted Sol‐gel Doped with Antimony‐Doped Tin Oxide
Authors: Lei Rulin , Guo Chunhui , Xiong Huayu , Dong Chao , Zhang Xiuhua , Wang Shengfu
Published: April 2014

A novel strategy to improve the sensitivity of molecularly imprinted polymer (MIP) sensors was proposed for the determination of β2‐agonists. The imprinted sol‐gel film was prepared by mixing silica sol with a functional monomer of antimony‐doped tin oxide (ATO) and a template of β2‐agonists. ATO, which was embedded in the surface of the molecularly imprinted sol‐gel film, not only provides the excellent conductivity for biosensor but also increases the stability and the surface area of the MIP film. The imprinted sensor was characterised by field emission scanning electron microscope, fourier transform infrared spectroscopy and electrochemical methods. Under the optimal experimental conditions, the peak current was linear with the logarithm of the concentration of clenbuterol (CLB) in the range of 5.5 nM–6.3 µM, and a detection limit of 1.7 nM was obtained. Meanwhile, the electrochemical sensor showed excellent specific recognition of the template molecule among structurally similar coexisting substances. Furthermore, the proposed sensor was satisfactorily applied to determine β2‐agonists in human serum samples. The good results indicated that highly effective molecularly imprinted sol‐gel films doped with ATO can be employed for other analytes.
Commonality in Liquidity Across International Borders: Evidence from Futures Markets
Authors: Frino Alex , Mollica Vito , Zhou Zeyang
Published: April 2014

This study examines commonality in liquidity for stock index futures markets. We report strong evidence of commonality in global liquidity for nine index futures contracts over a 10‐year time period extending October 2002 to September 2012. Our results are robust to expiry effects and tests for liquidity commonality based on a market model and principal component method. We investigate the variation in global liquidity commonality through time and document that liquidity commonality is higher in significance and more pervasive in recent years. © 2014 Wiley Periodicals, Inc. Jrl Fut Mark 34:807–818, 2014
Longitudinal reliability of tract‐based spatial statistics in diffusion tensor imaging
Authors: Madhyastha Tara , Mérillat Susan , Hirsiger Sarah , Bezzola Ladina , Liem Franziskus , Grabowski Thomas , Jäncke Lutz
Published: April 2014

Relatively little is known about reliability of longitudinal diffusion‐tensor imaging (DTI) measurements despite growing interest in using DTI to track change in white matter structure. The purpose of this study is to quantify within‐ and between session scan‐rescan reliability of DTI‐derived measures that are commonly used to describe the characteristics of neural white matter in the context of neural plasticity research. DTI data were acquired from 16 cognitively healthy older adults (mean age 68.4). We used the Tract‐Based Spatial Statistics (TBSS) approach implemented in FSL, evaluating how different DTI preprocessing choices affect reliability indices. Test‐Retest reliability, quantified as ICC averaged across the voxels of the TBSS skeleton, ranged from 0.524 to 0.798 depending on the specific DTI‐derived measure and the applied preprocessing steps. The two main preprocessing steps that we found to improve TBSS reliability were (a) the use of a common individual template and (b) smoothing DTI data using a 1‐voxel median filter. Overall our data indicate that small choices in the preprocessing pipeline have a significant effect on test‐retest reliability, therefore influencing the power to detect change within a longitudinal study. Furthermore, differences in the data processing pipeline limit the comparability of results across studies. Hum Brain Mapp 35:4544–4555, 2014. © 2014 Wiley Periodicals, Inc.
Movement‐related neuromagnetic fields in preschool age children
Authors: Cheyne Douglas , Jobst Cecilia , Tesan Graciela , Crain Stephen , Johnson Blake
Published: April 2014

We examined sensorimotor brain activity associated with voluntary movements in preschool children using a customized pediatric magnetoencephalographic system. A videogame‐like task was used to generate self‐initiated right or left index finger movements in 17 healthy right‐handed subjects (8 females, ages 3.2–4.8 years). We successfully identified spatiotemporal patterns of movement‐related brain activity in 15/17 children using beamformer source analysis and surrogate MRI spatial normalization. Readiness fields in the contralateral sensorimotor cortex began ∼0.5 s prior to movement onset (motor field, MF), followed by transient movement‐evoked fields (MEFs), similar to that observed during self‐paced movements in adults, but slightly delayed and with inverted source polarities. We also observed modulation of mu (8–12 Hz) and beta (15–30 Hz) oscillations in sensorimotor cortex with movement, but with different timing and a stronger frequency band coupling compared to that observed in adults. Adult‐like high‐frequency (70–80 Hz) gamma bursts were detected at movement onset. All children showed activation of the right superior temporal gyrus that was independent of the side of movement, a response that has not been reported in adults. These results provide new insights into the development of movement‐related brain function, for an age group in which no previous data exist. The results show that children under 5 years of age have markedly different patterns of movement‐related brain activity in comparison to older children and adults, and indicate that significant maturational changes occur in the sensorimotor system between the preschool years and later childhood. Hum Brain Mapp 35:4858–4875, 2014. © 2014 Wiley Periodicals, Inc.
Voxelwise lp‐ntPET for detecting localized, transient dopamine release of unknown timing: Sensitivity Analysis and Application to Cigarette Smoking in the PET Scanner
Authors: Kim Su Jin , Sullivan Jenna M. , Wang Shuo , Cosgrove Kelly P. , Morris Evan D.
Published: April 2014

The “linear parametric neurotransmitter PET” (lp‐ntPET) model estimates time variation in endogenous neurotransmitter levels from dynamic PET data. The pattern of dopamine (DA) change over time may be an important element of the brain's response to addictive substances such as cigarettes or alcohol. We have extended the lp‐ntPET model from the original region of interest (ROI) ‐ based implementation to be able to apply the model at the voxel level. The resulting endpoint is a dynamic image, or movie, of transient neurotransmitter changes. Simulations were performed to select threshold values to reduce the false positive rate when applied to real 11C‐raclopride PET data. We tested the new voxelwise method on simulated data, and finally, we applied it to 11C‐raclopride PET data of subjects smoking cigarettes in the PET scanner. In simulation, the temporal precision of neurotransmitter response was shown to be similar to that of ROI‐based lp‐ntPET (standard deviation ∼ 3 min). False positive rates for the voxelwise method were well controlled by combining a statistical threshold (the F‐test) with a new spatial (cluster‐size) thresholding operation. Sensitivity of detection for the new algorithm was greater than 80% for the case of short‐lived DA changes that occur in subregions of the striatum as might be the case with cigarette smoking. Finally, in 11C‐raclopride PET data, DA movies reveal for the first time that different temporal patterns of the DA response to smoking may exist in different subregions of the striatum. These spatiotemporal patterns of neurotransmitter change created by voxelwise lp‐ntPET may serve as novel biomarkers for addiction and/or treatment efficacy. Hum Brain Mapp 35:4876–4891, 2014. © 2014 The Authors. Human Brain Mapping Published by Wiley Periodicals, Inc.
Selenophene Electronics
Authors: Hollinger Jon , Gao Dong , Seferos Dwight S.
Published: April 2014

Substituting individual heavier (or lighter) atoms would appear to be an extremely straightforward method of controlling the optoelectronic properties of π‐conjugated molecules that has minimal impact on solid‐state materials properties, yet things are never as they appear. Selenophenes are a distinct class of material separate from thiophenes. Their HOMOLUMO gap is often narrow, which is attractive for photonic applications. Intermolecular SeSe interactions increase ordering on a molecular scale and lead to distinct solid‐state organization, which often leads to excellent charge‐transport properties. The crystallization of selenophene is distinct from thiophenes, and thus, composites of selenophenes and other organic materials have distinct nano‐ and microscale morphologies. Some of the best organic optoelectronic devices use selenophene‐containing materials, yet early results were less encouraging. The most recent syntheses, structure determination, and electronic device properties of π‐conjugated selenophene‐based materials are reviewed. Significantly more work is justified and this review sets the stage for those studies.