Journal of Molecular Liquids最新文献

{"title":"Ultrasound regulation of trypsin-enrofloxacin binding and conformational changes","authors":"Jiahao Li ,&nbsp;Shiwei Chen ,&nbsp;Chennan Zheng ,&nbsp;Yijin Chang ,&nbsp;Dazhi Zhou ,&nbsp;Fengming Li ,&nbsp;Changchun Hao","doi":"10.1016/j.molliq.2026.129264","DOIUrl":"10.1016/j.molliq.2026.129264","url":null,"abstract":"<div><div>This study investigates how ultrasound influences the interaction between trypsin (TRP) and enrofloxacin (ENRO) using integrated spectroscopic analysis and molecular simulations. The results show that ENRO quenches TRP fluorescence mainly through a static mechanism, with ultrasound enhancing the quenching effect and inducing more pronounced microenvironmental changes around Trp residues compared with thermal treatment. Circular dichroism analysis indicates that ultrasound leads to slight increases in β-sheet content and decreases in random coil, suggesting local structural ordering rather than unfolding. Molecular docking and dynamics simulations further confirm the stability of the TRP–ENRO complex and support the experimentally observed binding behavior. While the study provides molecular-level insight into how ultrasound modulates protein–ligand interactions, limitations remain, including the lack of enzymatic activity assays, cavitation characterization, and quantitative binding free-energy calculations. These findings establish a basis for future mechanistic and functional investigations.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"447 ","pages":"Article 129264"},"PeriodicalIF":5.2,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146102573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dimeric triazole-chalcone conjugate as multi-target directed β-amyloid inhibitor","authors":"Shivani Daksh ,&nbsp;Shashank Deep ,&nbsp;Anupama Datta","doi":"10.1016/j.molliq.2026.129327","DOIUrl":"10.1016/j.molliq.2026.129327","url":null,"abstract":"<div><div>The aggregation of the Aβ₄₂ protein and its interaction with metal ions are key contributors to the progression of neurodegenerative diseases, presenting significant challenges in developing effective therapeutic strategies. This study evaluated the triazole-chalcone conjugate, (<em>E</em>)-3-(4-(dimethylamino)phenyl)-1-(4-((1-(2-(4-((E)-3-(4(dimethylamino)phenyl) acryloyl)phenoxy)ethyl)-1H-1,2,3-triazol-4-yl)methoxy)phenyl)prop-2-en-1-one (L1), as a multi-target-directed ligand by assessing its Aβ₄₂ aggregation inhibition and disaggregation efficiency, metal-chelating ability, and its potential to attenuate metal-induced oxidative stress. Molecular docking studies revealed that L1 has a greater binding affinity towards Aβ₄₂ protofibril than the monomeric counterpart. Here, the two chalcone moieties were conjugated via click chemistry, employing a triazole linker known for its metal chelation properties and biological activity enhancement. In vitro thioflavin-T fluorescence assays demonstrated that L1 effectively inhibited self-induced Aβ₄₂ aggregation, achieving a maximum inhibition of 72.4 ± 2.3% at a 1:2 (Aβ₄₂:L1) molar ratio, with an IC₅₀ value of 3.19 ± 0.07 μM. Furthermore, L1 exhibited strong copper-chelating ability with a 2:1 (L1:Cu²⁺) stoichiometry and significantly attenuated Cu²⁺-induced Aβ₄₂ aggregation, displaying an IC₅₀ value of 5.41 ± 0.13 μM. In a Cu-ascorbate redox system, L1 markedly suppressed hydroxyl radical generation, indicating effective mitigation of metal-induced oxidative stress. Cellular studies using SH-SY5Y neuroblastoma cells confirmed the non-toxic nature of L1 and demonstrated substantial protection against Aβ₄₂-induced cytotoxicity. Collectively, these findings establish L1 as a promising multifunctional candidate that simultaneously inhibits Aβ₄₂ aggregation, disrupts preformed fibrils, chelates metal ions, and reduces oxidative stress, highlighting the therapeutic potential of dimeric triazole–chalcone conjugates for neurodegenerative disease intervention.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"447 ","pages":"Article 129327"},"PeriodicalIF":5.2,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146102574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Developments in green nanotechnology: biogenic synthesis, mechanisms, applications","authors":"Charu Agarwal ,&nbsp;Levente Csóka","doi":"10.1016/j.molliq.2026.129307","DOIUrl":"10.1016/j.molliq.2026.129307","url":null,"abstract":"<div><div>The conventional methods for fabrication of nanomaterials have heavily relied on the use of toxic chemicals for their reduction and stabilization as well as organic solvents, which are detrimental to the ecosystem. Thus, developing sustainable synthesis methods in line with the principles of “green” chemistry has always taken foremost precedence and at the same time, been a challenge. Over the past few decades, the synthesis of nanomaterials using environment-friendly agents has captivated the research community around the globe. A variety of plant extracts and microbes have been explored for the biogenic synthesis of different nanomaterials for myriad applications. This review aims to present the synthesis of nanomaterials using various green agents, their mechanism of formation and applications. Focus is also laid on how the morphology or property of the nanomaterials is influenced by different green agents or conditions during synthesis. Finally, the challenges and scope of using green agents for the synthesis of nanomaterials are presented.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129307"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146074571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"QSPR modeling of the thermal properties of alkylammonium mesylates based on quantum-chemical descriptors: gas-phase vs. CPCM calculations","authors":"I.V. Fedorova ,&nbsp;K.A. Fedorov ,&nbsp;L.P. Safonova","doi":"10.1016/j.molliq.2026.129328","DOIUrl":"10.1016/j.molliq.2026.129328","url":null,"abstract":"<div><div>The present study demonstrates that the thermal properties of alkylammonium mesylate protic ionic liquids (PILs) can be reliably predicted using quantitative structure-property relationship (QSPR) models constructed from a relatively small dataset and quantum-chemical descriptors obtained from dispersion-corrected density functional theory (DFT-D) calculations. Multiple linear regression models were developed to estimate decomposition and melting temperatures based on two descriptor sets: one derived from gas-phase calculations and the other obtained using the Conductor-like Polarizable Continuum Model (CPCM, dielectric constant ε = 24.85) to account for solvation effects. These descriptors capture both the structural features of cations and ion pairs and reflect the ion-ion interactions characteristic of PILs. The best QSPR models, incorporating five to eight descriptors, showed excellent agreement with experimental data, achieving coefficients of determination (R²) greater than 0.9 and root mean square errors (RMSE) bellow 12 °C. Models based on CPCM-derived descriptors reproduced experimental values more accurately than those relying on gas-phase descriptors. External validation using two additional PILs confirmed the robustness and practical applicability of the proposed models for estimating thermal properties within this class of ionic liquids.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129328"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146185157","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the interaction mechanism of the adsorption of red cabbage anthocyanins on silica from fossilized marine diatom rocks: adsorption equilibria modelling and DFT study","authors":"Jarrie P. Lima ,&nbsp;Adriana E. Costa-Sacchetto ,&nbsp;Toni J. Lopes ,&nbsp;Mara G.N. Quadri ,&nbsp;Marintho B. Quadri ,&nbsp;Erwin García-Hernández ,&nbsp;Adrian Bonilla-Petriciolet","doi":"10.1016/j.molliq.2026.129298","DOIUrl":"10.1016/j.molliq.2026.129298","url":null,"abstract":"<div><div>The separation of bioactive compounds, such as anthocyanins, is of considerable interest for their incorporation into chemical and industrial formulations. In this study, the adsorption behavior of anthocyanins (ANT) extracted from red cabbage onto a silica from fossilized marine diatom rocks (SAD), derived from fossilized marine diatomaceous earth, was systematically investigated. Batch adsorption experiments were conducted under different operational conditions, and the equilibrium data were analyzed using classical isotherm models as well as advanced multilayer statistical physics modeling. In addition, density functional theory (DFT) calculations were employed to elucidate the molecular-level interactions governing the adsorption process. The results demonstrated that acidic conditions (pH 3) favor ANT adsorption onto SAD. The adsorption capacity ranged from 31.28 ± 0.5 to 16.91 ± 0.9 mg g⁻¹ within the temperature range of 298–328 K. The Freundlich model adequately described the experimental equilibrium data, while the multilayer model provided deeper insight into the adsorption mechanism. Both experimental characterization and DFT simulations indicated that hydrogen bonding interactions, with an interaction energy of approximately −2.9 eV, play a dominant role in ANT adsorption. Thermodynamic analysis confirmed that the adsorption process is spontaneous and exothermic. Furthermore, SAD exhibited good regeneration performance, maintaining its adsorption capacity over up to seven adsorption–desorption cycles. These findings highlight the potential of SAD as an efficient and reusable adsorbent for the recovery of bioactive compounds in liquid-phase systems relevant to industrial applications.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129298"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146185186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unravelling structure and dynamics of pyrrolidinium based amino acid ionic liquids for CO2 capture: A combined DFT and MD study","authors":"Khusboo Dubey,&nbsp;Raghu Nath Behera","doi":"10.1016/j.molliq.2026.129335","DOIUrl":"10.1016/j.molliq.2026.129335","url":null,"abstract":"<div><div>Amino acid based ionic liquids (AAILs), and in general ionic liquids, are versatile solvents for numerous applications, including gas capture. Here, we have investigated three AAILs ([Pyr][His], [Pyr][Lys], and [Pyr][Arg]) consisting of N-butyl, N-methyl pyrrolidinium [Pyr] cation with Lysine [Lys], Histidine [His], and Arginine [Arg] anions for CO₂ capture. The structure and interactions in neat AAILs and in the presence of CO₂ (AAIL-CO₂ complexes), and the mechanism of CO₂ absorption are investigated with DFT calculations along with Atoms-in-Molecules (AIM) analysis. The hydrogen bond interactions observed between the cation-anion pairs are predominantly electrostatic in nature, with moderate to weak strength. The HOMO-LUMO gaps of the AAILs are found to increase with CO₂ absorption. The [Pyr][His] system exhibits the lowest activation energy barrier for the CO₂ absorption reaction. Further insights into CO₂ binding, transport properties, and hydrogen-bond dynamics are obtained from classical molecular dynamics simulations. The density profile curves indicated that the CO₂ molecules are most likely accumulated at the CO₂/AAILs interfacial region, following the trend [Pyr][His] &gt; [Pyr][Arg] &gt; [Pyr][Lys]. The time-correlation functions for hydrogen bonds have been calculated to analyse the dynamic properties. The self-diffusion coefficients of ions increase, while the bulk densities of AAILs remain almost constant upon CO₂ binding. The [Pyr][Lys] system showed faster dynamics consistent with higher diffusion and ionic conductivity.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129335"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146185187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Promising natural compounds for hypoglycemic therapy: in silico identification of GKRP inhibitors from Coffea arabica via DFT, molecular docking, and aqueous-phase molecular dynamics simulations","authors":"Jonathan Jason Sugihcahyana ,&nbsp;Christian Kenneth ,&nbsp;Dwi Hudiyanti ,&nbsp;Parsaoran Siahaan ,&nbsp;Daru Seto Bagus Anugrah","doi":"10.1016/j.molliq.2026.129256","DOIUrl":"10.1016/j.molliq.2026.129256","url":null,"abstract":"<div><div>Diabetes mellitus remains a significant global health problem, and targeting glucokinase regulatory protein (GKRP) offers a promising strategy to regulate glucokinase (GK) activity more safely without increasing the risk of hypoglycemia. Coffee, which is rich in bioactive phenolic compounds, has the potential to provide a natural product-based antidiabetic candidate. This study aims to identify <em>Coffea arabica</em> compounds as potential GKRP inhibitors and to characterize the complexes' dynamics. Initially, <em>Coffea arabica</em> compounds were screened using a blind docking approach against GKRP and ADME filtering. Out of 134 compounds, 12 compounds, particularly Fontanesine B (Fon), Formononetin (For), and Hesperetin (Hes), exhibited high binding scores (−8.1 to −10.6 kcal/mol) while adhering to Lipinski's rule and showing high gastrointestinal absorption. The 12 potential compounds were further characterized using density functional theory (DFT), simulated with GKRP-F1P complex in triplicate 200 ns molecular dynamics (MD) simulations, and calculated for their binding free energy using MM/GBSA. Principal component analysis (PCA) showed that some coffee ligands restrict the collective motion of GKRP, while the dynamic cross-correlation matrix (DCCM) revealed a strengthening of the motion correlation around the F1P binding site and the GK binding interface. Most of the ligands are strong electrophiles that interact with the key nucleophilic residue Glu153, stabilizing the bound configuration of F1P and lowering its binding energy. Overall, these results highlight coffee-derived polyphenols as promising GKRP inhibitors that can potentially maintain glucokinase activity and provide a basis for the development of coffee-based antidiabetic candidates.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129256"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146035898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human serum albumin: Amyloid or amorphous aggregation? The influence of thiol status and ligand binding","authors":"Natalija Andrejević ,&nbsp;Predrag Petrović ,&nbsp;Sanja Stevanović ,&nbsp;Tomaž Urbič ,&nbsp;Natalija Polović","doi":"10.1016/j.molliq.2026.129290","DOIUrl":"10.1016/j.molliq.2026.129290","url":null,"abstract":"<div><div>Under destabilizing conditions, proteins are prone to misfold and aggregate into disordered amorphous aggregates or highly organized amyloid fibrils. Human serum albumin (HSA) is an abundant protein in human plasma and plays a key role in maintaining redox balance due to its high disulfide bond content. It also serves as a carrier for a great number of ligands, including fatty acids. Given its physiological importance, understanding HSA's aggregation behavior is crucial.</div><div>This study investigates how ligand binding and redox state influence HSA aggregation under destabilizing conditions. Prolonged thermal treatment in acidic conditions was applied to destabilize the protein and to compare aggregation type in ligand-saturated and ligand-free HSA, before and after mild reduction with dithiothreitol (DTT). Structural changes and aggregate formation were monitored using infrared spectroscopy, dynamic light scattering, atomic force microscopy, and fluorescent dye assays (Thioflavin T and 8-anilinonaphthalene-1-sulfonic acid). Our results showed that reduction did not significantly alter HSA's native structure but influenced its aggregation pathway. In reduced, ligand-free HSA, thermal treatment enhanced fibrillation potential, whereas in reduced ligand-saturated HSA, thermal treatment mainly resulted in amorphous aggregates. These findings suggest that the presence of ligands, especially under reduced conditions, shifts aggregation from amyloid fibrils to amorphous aggregates. To further confirm this, ligand-saturated HSA prepared with palmitic acid showed reduced fibrillation potential, supporting the hypothesis that ligand-presence along with free thiol content modulate aggregation behavior.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129290"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146035961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of H/D isotope substitution on the structure-packing and interaction-related properties of water as a solute in tetramethylurea at T = (278.15 to 318.15) K and p = 0.1 MPa","authors":"Evgeniy V. Ivanov","doi":"10.1016/j.molliq.2026.129318","DOIUrl":"10.1016/j.molliq.2026.129318","url":null,"abstract":"<div><div>Densities of solutions of ordinary (H₂O) and heavy (D₂O) water in tetramethylurea (TMU) were measured with an uncertainty of ≤0.05 kg∙m⁻³ at <em>T</em> = (278.15, 288.15, 298.15, 308.15, and 318.15) K under ambient pressure using a precision densimeter equipped with the oscillating U-shaped tube. The solution molality, <em>m</em>, was ranged from (0.055 to 1.16) mol·(kg TMU)⁻¹ in both H/D isotopically distinguishable water-containing systems. Standard molar volumes, <span><math><msubsup><mover><mi>V</mi><mo>¯</mo></mover><mi>w</mi><mi>o</mi></msubsup><mfenced><mrow><mo>≡</mo><msubsup><mover><mi>V</mi><mo>¯</mo></mover><mi>w</mi><mo>∞</mo></msubsup></mrow></mfenced></math></span>, and isobaric expansibilities, <span><math><msubsup><mover><mi>E</mi><mo>¯</mo></mover><mrow><mi>p</mi><mo>,</mo><mi>w</mi></mrow><mi>o</mi></msubsup><mfenced><mrow><mo>≡</mo><msubsup><mover><mi>E</mi><mo>¯</mo></mover><mrow><mi>p</mi><mo>,</mo><mi>w</mi></mrow><mo>∞</mo></msubsup></mrow></mfenced></math></span>, for H₂O and D₂O as solutes in TMU were computed. The temperature-dependent behavior of the solute D₂O–H₂O isotope effects (IEs) in the standard (partial) volume properties of water in TMU was discussed taking account of structure-packing and interaction-related peculiarities of the solvation complex formed. The factors influencing the structure state of water H/D isotopologues dissolved in TMU were considered. Attention was also paid to correlations between the currently available IEs of interest for water-containing aprotic protophilic media including TMU and some interaction-related properties of the latter.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129318"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146185185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photoactive thermotropic liquid crystal systems and asymmetric electrochemical polymerization","authors":"Aoi Tokutake,&nbsp;Hiromasa Goto","doi":"10.1016/j.molliq.2026.129295","DOIUrl":"10.1016/j.molliq.2026.129295","url":null,"abstract":"<div><div>Photoactive-chiral liquid crystal—a photoactive dopant, a chiral dopant, and a smectic C* (SmC*)–inducing dopant were developed. Upon UV irradiation, the liquid-crystal system transformed into an isotropic phase, which subsequently reverted to a liquid-crystal phase under visible (Vis) light. In other words, Vis light generated the mesophase. Electrochemical polymerization conducted in chiral nematic (cholesteric, N*), smectic A, and SmC* phases yielded aggregation-imprinted, electro-optically active conjugated polymers. Electrochemically driven changes in the optical activity of the resulting polymers were confirmed using circular dichroism optical absorption measurements.</div></div>","PeriodicalId":371,"journal":{"name":"Journal of Molecular Liquids","volume":"446 ","pages":"Article 129295"},"PeriodicalIF":5.2,"publicationDate":"2026-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146074431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}