bluuebunny/crossref_metadata_2025_split huggingface dataset:
Statistical vs. Stochastic experimental design: An experimental comparison on the example of protein refolding
Authors: Anselment Bernd , Schoemig Veronika , Kesten Christopher , Weuster‐Botz Dirk 
Published: September 2012

AbstractOptimization of experimental problems is a challenging task in both engineering and science. In principle, two different design of experiments (DOE) strategies exist: statistical and stochastic methods. Both aim to efficiently and precisely identify optimal solutions inside the problem‐specific search space. Here, we evaluate and compare both strategies on the same experimental problem, the optimization of the refolding conditions of the lipase fromThermomyces lanuginosuswith 26 variables under study. Protein refolding is one of the main bottlenecks in the process development for recombinant proteins. Despite intensive effort, the prediction of refolding from sequence information alone is still not applicable today. Instead, suitable refolding conditions are typically derived empirically in large screening experiments. Thus, protein refolding should constitute a good performance test for DOE strategies. We compared an iterative stochastic optimization applying a genetic algorithm and a standard statistical design consisting of a D‐optimal screening step followed by an optimization via response surface methodology. Our results revealed that only the stochastic optimization was able to identify optimal refolding conditions (∼1.400 U g−1refolded activity), which were 3.4‐fold higher than the standard. Additionally, the stochastic optimization proved quite robust, as three independent optimizations performed similar. In contrast, the statistical DOE resulted in a suboptimal solution and failed to identify comparable activities. Interactions between process variables proved to be pivotal for this optimization. Hence, the linear screening model was not able to identify the most important process variables correctly. Thereby, this study highlighted the limits of the classic two‐step statistical DOE. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012
Live‐Cell Studies of p300/CBP Histone Acetyltransferase Activity and Inhibition
Authors: Dancy Beverley M. , Crump Nicholas T. , Peterson Daniel J. , Mukherjee Chandrani , Bowers Erin M. , Ahn Young‐Hoon , Yoshida Minoru , Zhang Jin , Mahadevan Louis C. , Meyers David J. , Boeke Jef D. , Cole Philip A. 
Published: September 2012

AbstractHistone acetyltransferase enzymes (HATs) are important therapeutic targets, but there are few cell‐based assays available for evaluating the pharmacodynamics of HAT inhibitors. Here we present the application of a FRET‐based reporter, Histac, in live‐cell studies of p300/CBP HAT inhibition, by both genetic and pharmacologic disruption. shRNA knockdown of p300/CBP led to increased Histac FRET, thus suggesting a role for p300/CBP in the acetylation of the histone H4 tail. Additionally, we describe a new p300/CBP HAT inhibitor, C107, and show that it can also increase cellular Histac FRET. Taken together, these studies provide a live‐cell strategy for identifying and evaluating p300/CBP inhibitors.
Crystal Structures of BapA Complexes with β‐Lactam‐Derived Inhibitors Illustrate Substrate Specificity and Enantioselectivity of β‐Aminopeptidases
Authors: Heck Tobias , Merz Tobias , Reimer Artur , Seebach Dieter , Rentsch Daniel , Briand Christophe , Grütter Markus G. , Kohler Hans‐Peter E. , Geueke Birgit 
Published: September 2012

Abstractβ‐Aminopeptidases have exclusive biocatalytic potential because they react with peptides composed of β‐amino acids, which serve as building blocks for the design of non‐natural peptidomimetics. We have identified the β‐lactam antibiotic ampicillin and the ampicillin‐derived penicilloic acid as novel inhibitors of the β‐aminopeptidase BapA from Sphingosinicella xenopeptidilytica (Ki values of 0.69 and 0.74 mM, respectively). We report high‐resolution crystal structures of BapA in noncovalent complexes with these inhibitors and with the serine protease inhibitor 4‐(2‐aminoethyl)benzenesulfonyl fluoride. All three inhibitors showed similar binding characteristics; the aromatic moiety extended into a hydrophobic binding pocket of the active site, and the free amino group formed a salt bridge with Glu133 of BapA. The exact position of the inhibitors and structural details of the ligand binding pocket illustrate the specificity and the enantioselectivity of BapA‐catalyzed reactions with β‐peptide substrates.
CO2‐Fixation on Aliphatic α,ω‐Diamines to Form Cyclic Ureas, Catalyzed by Ceria Nanoparticles that were Obtained by Templating with Alginate
Authors: Primo Ana , Aguado Eric , Garcia Hermenegildo 
Published: September 2012

AbstractCeria nanoparticles (average particle size: 8 nm) have been obtained by the calcination of alginate aerogel beads that were precipitated from aqueous solutions of (NH4)2Ce(NO3)6. These nanoparticles were considerably more active as a catalyst for CO2‐insertion into aliphatic α,ω‐diamines than the analogous commercial CeO2 with larger particle size (40 nm). CeO2 that was obtained by templating with the natural alginate biopolymer afforded the cyclic urea of ethylenediamine in EtOH solvent at 160 °C in 37 % yield. This yield is remarkable for a process that involves CO2 as a feedstock. Other α,ω‐diamines, such as diethylenetriamine, N,N′‐dimethylethylenediamine, N‐(2‐aminoethyl)acetamide, and 1,4‐diaminobutane, also formed their corresponding cyclic ureas in 4–36 % yield. The catalyst lost activity upon reuse, thereby leading to severe deactivation that was only partially recovered by washing with aqueous acidic solutions.
Highly Selective Linear Dimerization of Styrenes by Ceria‐Supported Ruthenium Catalysts
Authors: Shimura Shun , Miura Hiroki , Tsukada Shinji , Wada Kenji , Hosokawa Saburo , Inoue Masashi 
Published: September 2012

AbstractCeria‐supported ruthenium (Ru/CeO2) was found to be an effective catalyst for linear‐selective dimerization of styrenes. The dimerization of styrene in the presence of Ru/CeO2 together with ethanol and formalin in mesitylene at 150 °C gave (E)‐1,4‐diaryl‐1‐butene in 63 % yield with high linear selectivity (95 %). The presence of ethanol and/or formaldehyde is indispensable. Whereas a zirconia‐supported ruthenium showed moderate activity, catalysts supported on other metal oxides were not effective at all. The hot filtration tests indicated that the Ru/CeO2 catalyst acts heterogeneously. The leaching of ruthenium species into the solution was negligible according to analysis by using inductively coupled plasma atomic emission spectroscopy. The regioselectivity can be switched through the choice of a suitable solvent: the reactions in N,N‐dimethylacetamide or N‐methyl‐2‐pyrrolidone gave the corresponding branched dimers in good yields with complete regioselectivity.
Synthesis and Temperature‐Induced Morphological Control in a Hybrid Porous Iron–Phosphonate Nanomaterial and Its Excellent Catalytic Activity in the Synthesis of Benzimidazoles
Authors: Dutta Arghya , Mondal John , Patra Astam K. , Bhaumik Asim 
Published: September 2012

AbstractA new organic–inorganic hybrid porous iron–phosphonate material, HPFP‐1, has been synthesized under hydrothermal conditions by using hexamethylenediamine‐N,N,N′,N′‐tetrakis‐(methylphosphonic acid) (HDTMP) as the organophosphorus precursor. The morphology of this material was found to be different at three different temperatures. The material that was synthesized at 453 K showed a flake‐like particle morphology and the material was highly crystalline. Whereas, the materials that were synthesized at 443 K and 423 K were semi‐crystalline and showed rod‐like‐ and spherical morphological features, respectively. SEM and TEM were employed to understand this change in particle morphology depending on the reaction temperature. Powder XRD analysis suggested the formation of a new tetragonal phase in HPFP‐1 (a=11.313, c=15.825 Å; V=2025.659 Å3). N2‐sorption analysis suggested the existence of supermicropores and interparticle mesopores in these materials. Elemental‐ and thermal analyses, as well as FTIR spectroscopy, were employed to verify the composition and framework bonding of the material. The HPFP‐1 material showed excellent catalytic activity for the synthesis of benzimidazole derivatives under mild liquid‐phase reaction conditions.
Controlling the Assembly of Chalcogenide Anions in Ionic Liquids: From Binary Ge/Se through Ternary Ge/Sn/Se to Binary Sn/Se Frameworks
Authors: Lin Yumei , Massa Werner , Dehnen Stefanie 
Published: September 2012

AbstractSeven compounds with binary or ternary Ge/Se, Ge/Sn/Se, or Sn/Se anionic substructures crystallized upon the ionothermal reactions of [K4(H2O)3][Ge4Se10] with SnCl4⋅5 H2O or SnCl2 in [BMMIm][BF4] or [BMIm][BF4] (BMMIm=1‐butyl‐2,3‐dimethyl‐imidazolium, BMIm=1‐butyl‐3‐methyl‐imidazolium). The products were obtained by subtly varying the reaction conditions; the nature and amount of an additional amine was the most important parameter in the product selection and in determining the Sn/Ge ratio in the isolated products. The crystal structures of these chalcogenides were based on complex anions with unprecedented topologies that varied from discrete clusters (0D) through 1D chain structures or 2D layers to 3D frameworks. The architecture and composition of the title compounds were well reflected by their optical absorption behavior. Herein, we report a convenient approach for the generation of chalcogenidometallate phases with fine‐tunable electronic properties in ionic liquids, which have been inaccessible by traditional methods.
ChemInform Abstract: Well‐Defined Crystallites Autoclaved from the Nitrate/NH4OH Reaction System as the Precursor for (Y,Eu)2O3 Red Phosphor: Crystallization Mechanism, Phase and Morphology Control, and Luminescent Property.
Authors: Zhu Qi , Li Ji‐Guang , Ma Renzhi , Sasaki Takayoshi , Yang Xiaojing , Li Xiaodong , Sun Xudong , Sakka Yoshio 
Published: September 2012

AbstractHexagonal platelets of Ln2(OH)5NO3·nH2O (Ln: Y, Eu), hexagonal prisms and microwires of Ln4O(OH)9NO3, square nanoplatelets of Ln2.94(NO3)0.06·nH2O, and spindle‐like microrods of Ln(OH)3 are hydrothermally synthesized from aqueous solutions of Ln(NO3)3 (sometimes together with NH4NO3) and NH4OH (autoclave, pH 6—13, 120—200 °C, 24 h).
ChemInform Abstract: Syntheses, Structures, and Optical Properties of Ba4MInSe6 (M: Cu, Ag).
Authors: Yin Wenlong , Feng Kai , Hao Wenyu , Yao Jiyong , Wu Yicheng 
Published: September 2012

AbstractThe new title compounds are synthesized by solid state reactions of M2Se (M: Cu, Ag), BaSe, and In2Se3 (973—1073 K, 48 h).
ChemInform Abstract: Mononuclear Palladium and Heterodinuclear Palladium—Ruthenium Complexes of Semicarbazone Ligands. Synthesis, Characterization, and Application in C—C Cross‐Coupling Reactions.
Authors: Datta Sayanti , Seth Dipravath Kumar , Halder Sarmistha , Sheldrick William S. , Mayer‐Figge Heike , Drew Michael G. B. , Bhattacharya Samaresh 
Published: September 2012

AbstractA variety of mononuclear palladium and heterobinuclear Pd—Ru N,N,O‐pincer complexes is synthesized and studied in Suzuki, Heck and Sonogashira cross‐coupling.
ChemInform Abstract: Recyclable Hypervalent Iodine(III) Reagent Iodosodilactone as an Efficient Coupling Reagent for Direct Esterification, Amidation, and Peptide Coupling.
Authors: Tian Jun , Gao Wen‐Chao , Zhou Dong‐Mei , Zhang Chi 
Published: September 2012

AbstractUnder these conditions a wide range of esters, amides, and peptides as well as macrocyclic lactones can be prepared in good to high yields.
ChemInform Abstract: Synthesis of 1,2‐Disubstituted Cyclopentenes by Palladium‐Catalyzed Reaction of Homopropargyl‐Substituted Dicarbonyl Compounds with Organic Halides via 5‐Endo‐Dig Cyclization.
Authors: Fujino Daishi , Yorimitsu Hideki , Osuka Atsuhiro 
Published: September 2012

AbstractAromatic, alkenyl‐ and alkynyl‐ bromides and chlorides are used affording cyclopentene products with good yields.
ChemInform Abstract: Asymmetric Organocatalytic Cascade Michael/Michael/Henry Reaction Sequence: Control of All Stereocenters in One Cyclohexane Skeleton.
Authors: Mao Zhifeng , Jia Yaomei , Xu Zhaoqing , Wang Rui 
Published: September 2012

AbstractHexasubstituted chiral cyclohexanols are constructed in one step.
ChemInform Abstract: Control of Six Contiguous Stereocenters in an Asymmetric Organocatalytic One‐Pot Michael/Michael/Aldol Addition Sequence.
Authors: Enders Dieter , Urbanietz Gregor , Cassens‐Sasse Elisa , Keess Sebastian , Raabe Gerhard 
Published: September 2012

AbstractThe reaction of β‐ketoesters, aromatic nitroalkenes and aromatic α,β‐unsaturated aldehydes provides highly substituted cyclohexanes with 6 contiguous stereocenters including a quaternary one.
ChemInform Abstract: Rhodium‐Catalyzed Allylic Substitution Reactions with Indium(III) Organometallics.
Authors: Riveiros Ricardo , Tato Ruben , Perez Sestelo Jose , Sarandeses Luis A. 
Published: September 2012

AbstractOnly in the case of (IVc) a slight amount of the γ‐adduct is obtained due to steric effects.
ChemInform Abstract: One‐Pot Primary Aminomethylation of Aryl and Heteroaryl Halides with Sodium Phthalimidomethyltrifluoroborate.
Authors: Murai Norio , Miyano Masayuki , Yonaga Masahiro , Tanaka Keigo 
Published: September 2012

AbstractA one‐pot primary aminomethylation with substrate (I) leads to the corresponding amines and is followed by deamidination to afford the desired products.
ChemInform Abstract: Knoelker′s Iron Complex: An Efficient in situ Generated Catalyst for Reductive Amination of Alkyl Aldehydes an Amines.
Authors: Pagnoux‐Ozherelyeva Anastassiya , Pannetier Nicolas , Mbaye Mbaye Diagne , Gaillard Sylvain , Renaud Jean‐Luc 
Published: September 2012

AbstractThe first homogeneous well‐defined iron catalyst for the title reaction under low pressure of hydrogen is reported.
ChemInform Abstract: A Novel Multicomponent Tandem Phosphine‐Catalyzed Umpolung Reaction: Facile Access to Highly Functionalized α‐Aminonitriles.
Authors: Chen Jian‐Ming , Fang Yin‐Zhi , Wei Zhong‐lin , Liao Wei‐Wei 
Published: September 2012

AbstractBenzylamine and morpholine do not react and substituted allenes are less suitable than (IV).
ChemInform Abstract: The Vinylogous Aldol Reaction of Unsaturated Esters and Enolizable Aldehydes Using the Novel Lewis Acid Aluminum Tris(2,6‐di‐2‐naphthylphenoxide).
Authors: Gazaille Jeffrey A. , Sammakia Tarek 
Published: September 2012

AbstractAcid (I) generated in situ, promotes the reaction with various aldehydes including hindered, α‐branched and oxygenated ones.
ChemInform Abstract: Efficient Synthesis of γ‐Keto Sulfones by NHC‐Catalyzed Intermolecular Stetter Reaction.
Authors: Bhunia Anup , Yetra Santhivardhana Reddy , Bhojgude Sachin Suresh , Biju Akkattu T. 
Published: September 2012

AbstractThe reaction is applied to a wide range of aromatic, heteroaromatic and aliphatic aldehydes.
ChemInform Abstract: Three‐Component Synthesis of Substituted β‐(Trifluoromethyl)pyrroles via Grob Cyclization of 1,1,1‐Trifluoro‐3‐nitrobut‐2‐ene with 1,3‐Dicarbonylic Compounds and Ammonia or Primary Amines.
Authors: Korotaev Vladislav Yu. , Barkov Alexey Yu. , Kotovich Ivan V. , Sosnovskikh Vyacheslav Ya. 
Published: September 2012

AbstractA variety of substituted β‐(trifluoromethyl)pyrroles of type (IV) is prepared via a one‐pot three‐component Grob cyclization of 1,1,1‐trifluoro‐3‐nitrobut‐2‐ene (I) with 1,3‐dicarbonylic compounds and ammonia or primary amines.
ChemInform Abstract: A New Tandem Cross Metathesis—Intramolecular Aza‐Michael Reaction for the Synthesis of α,α‐Difluorinated Lactams.
Authors: Fustero Santos , Baez Claribel , Sanchez‐Rosello Maria , Asensio Amparo , Miro Javier , del Pozo Carlos 
Published: September 2012

AbstractDifluorinated amides are treated with methyl vinyl ketone or analogous esters to produce the desired γ‐ and δ‐lactam units including optically active versions.
ChemInform Abstract: Asymmetric Pericyclic Cascade Approach to Spirocyclic Oxindoles.
Authors: Richmond Edward , Duguet Nicolas , Slawin Alexandra M. Z. , Lebl Tomas , Smith Andrew D. 
Published: September 2012

AbstractThe reaction of racemic nitrones with cycloalkylketenes affords the spirocyclic oxindoles in good yields.
ChemInform Abstract: A Highly Efficient Cu‐Catalyst System for N‐Arylation of Azoles in Water.
Authors: Wang Deping , Zhang Fuxing , Kuang Daizhi , Yu Jiangxi , Li Junhua 
Published: September 2012

AbstractThe combination of CuI, an oxime, and Bu4NI is successfully applied as a catalytic system for the N‐arylation of imidazoles (I) and (VI), benzimidazole (IV), pyrazole (IX), and triazole (XII).
ChemInform Abstract: Iron‐Catalyzed 2‐Arylbenzoxazole Formation from o‐Nitrophenols and Benzylic Alcohols.
Authors: Wu Mingyue , Hu Xiong , Liu Juan , Liao Yunfeng , Deng Guo‐Jun 
Published: September 2012

AbstractThe reaction of nitrophenols with benzylic and heterobenzylic alcohols tolerates a wide range of functional groups.
ChemInform Abstract: Synthesis and anti‐Norovirus Activity of Pyranobenzopyrone Compounds.
Authors: Pokhrel Laxman , Kim Yunjeong , Nguyen Thi D. T. , Prior Allan M. , Lu Jianyu , Chang Kyeong‐Ok , Hua Duy H. 
Published: September 2012

AbstractA series of 14 title compounds of type (IV) is tested for anti‐norovirus effects.
ChemInform Abstract: Synthesis of Indolizidinone Analogues of Cytotoxic Alkaloids: Monocyclic Precursors Are also Active.
Authors: Boto Alicia , Miguelez Javier , Marin Raquel , Diaz Mario 
Published: September 2012

AbstractA two‐step route is reported for the transformation of readily available hydroxyproline N‐amides into indolizidinone derivatives of type (V) and (VI).
ChemInform Abstract: Ultrasound‐Assisted Synthesis and Preliminary Bioactivity of Novel 2H‐1,2,4‐Thiadiazolo[2,3‐a]pyrimidine Derivatives Containing Fluorine.
Authors: Wang Mao Rong , Jiang Lin , Zhou Shao Fang , Zhang Ze Yuan , Ji Zeng Chen 
Published: September 2012

AbstractThe four‐step synthesis of the title compounds (III) is based on the ultrasound‐ promoted oxidative cyclization of precursors (II).
ChemInform Abstract: Synthesis of Alkaloid‐Like Compounds via the Bridging Ritter Reaction.
Authors: Hemtasin Chulida , Ung Alison T. , Kanokmedhakul Somdej , Kanokmedhakul Kwanjai , Bishop Roger , Satraruji Thanaphat , Bishop David 
Published: September 2012

Abstract10 Alkaloid‐like compounds are prepared using alcohol (I) as starting product.
ChemInform Abstract: Palladium‐Catalyzed Desymmetrization of Silacyclobutanes with Alkynes: Enantioselective Synthesis of Silicon‐Stereogenic 1‐Sila‐2‐cyclohexenes and Mechanistic Considerations.
Authors: Shintani Ryo , Moriya Kohei , Hayashi Tamio 
Published: September 2012

AbstractEnantioenriched products are obtained from the title reactions.
ChemInform Abstract: Efficient Synthesis of α‐Tertiary α‐Silylamines from Aryl Sulfonylimidates via One‐Pot, Sequential C—Si/C—C Bond Formations.
Authors: Han Xiao‐Jun , Yao Ming , Lu Chong‐Dao 
Published: September 2012

AbstractBulky Grignard reagents possessing a β‐hydrogen atom react poorly with formation of an α‐branched α‐silylamine (XI) as side product, generated via hydride transfer.
ChemInform Abstract: Polycyclic Polyprenylated Acylphloroglucinols and Cytotoxic Constituents of Hypericum androsaemum.
Authors: Wang Kou , Wang Yuan‐Yuan , Gao Xiu , Chen Xuan‐Qin , Peng Li‐Yan , Li Yan , Xu Gang , Zhao Qin‐Shi 
Published: September 2012

AbstractAndroforin A (I) and hyperandrone A (II) are isolated from the aerial parts together with a series of known constituents.
ChemInform Abstract: 1,3‐Dipolar Cycloadditions of Nitrones to Hetero‐Substituted Alkenes. Part 2. Sila‐, Thia‐, Phospha‐ and Halo‐Substituted Alkenes
Authors: Nguyen Thanh Binh , Martel Arnaud , Gaulon‐Nourry Catherine , Dhal Robert , Dujardin Gilles 
Published: September 2012

AbstractReview: 139 refs.
Mortality Rate Increases Steeply With Nonadherence to Statin Therapy in Patients With Acute Coronary Syndrome
Authors: Allonen Jaakko , Nieminen Markku S. , Lokki Maisa , Parkkonen Olavi , Vaara Satu , Perola Markus , Hiekkalinna Tero , Strandberg Timo E. , Sinisalo Juha 
Published: September 2012

AbstractBackground:In a prospective cohort of consecutive acute coronary syndrome (ACS) patients, we compared the adherence rate of statin usage and mortality rate during a median follow‐up of 23 months.Hypothesis:Adherence to statin therapy after acute coronary syndrome affects mortality rate.Methods:We analyzed ACS patients (N = 1969; age, 65.9 ± 11.8 years; female 30.4%) who underwent angiography between March 2006 and March 2008. The postdischarge usage of statins was based on the purchase register of the Social Insurance Institution of Finland. The death rate was verified from Statistics Finland.Results:At discharge, the rate of statin prescription to patients was 95.4% (n = 1878). When comparing adherent patients (n = 1200; 61.7%), who purchased the medication systematically until the end of the median 23‐month follow‐up, with nonadherent patients (n = 94; 4.8%), who did not use the medication at all, there was a vast difference in absolute death rate between the groups: 4.9% vs 14.9%, respectively (P < 0.001). We conducted Cox proportional hazards model with ACS type, cerebrovascular attack, diabetes, age, 3‐artery disease, and cancer as adjusted confounders. Compared with regular statin users, nonusers were associated with a >2× increased hazard ratio of mortality (hazard ratio: 2.70, 95% confidence interval: 1.49‐4.90, P = 0.001).Conclusions:Statin medication is essential for discharged ACS patients. They should be strongly encouraged to purchase and use it.The authors have no funding, financial relationships, or conflicts of interest to disclose.
Litter Decomposition of Six Macrophytes in a Eutrophic Shallow Lake (Baiyangdian Lake, China)
Authors: Lan Yan , Cui Baoshan , You Zheyuan , Li Xia , Han Zhen , Zhang Yongtao , Zhang Yu 
Published: September 2012

AbstractThe decomposition of aquatic macrophytes has important consequences for wetlands because it is closely related to organic matter accumulation and nutrient cycling. A field litter bag experiment was undertaken to investigate the decomposition rates of six dominant macrophytes. And the level of nutrient transfer from plant residues to lake water was also researched in Baiyangdian Lake, a typical macrophyte‐dominated lake in the Northern China Plain. Intact standing plants of Phragmites australis, Typha angustifolia, Nelumbo nucifera, Potamogeton pectinatus, Ceratophyllum demersum, and Chara sp. were collected and different plant tissues incubated near sediment traps. Litter bags were collected at days 6, 26, 60, 104, and 140 to follow biomass loss and nutrient release. Decomposition rates of different macrophytes calculated from an exponential model ranged from 0.0028 to 0.0110 day−1. For the same external environmental conditions, decomposition rates were significantly affected by initial litter chemistry. The remaining biomass was positively related to the C/N and C/P ratios and negatively related to the N and P concentrations and the N/P ratio. Both stems and roots showed a lower decomposition rate than the leaves of the same species due to the higher C/N ratio. The nutrient release from plant litter to the lake water was substantial. Over the incubation period, the release of N from the litter represented 18.10–88.79% of the initial content, and the release of P from the litter represented 24.26–92.83% of the initial content.