Journal of Inorganic Biochemistry最新文献

{"title":"Structure, properties, and decomposition in biological systems of a new nitrosyl iron complex with 2-methoxythiophenolyls, promising for the treatment of cardiovascular diseases","authors":"Оlesya V. Pokidova ,&nbsp;Veronika O. Novikova ,&nbsp;Nina S. Emel’yanova ,&nbsp;Ludmila M. Mazina ,&nbsp;Alina S. Konyukhova ,&nbsp;Alexander V. Kulikov ,&nbsp;Gennadii V. Shilov ,&nbsp;Nikolai S. Ovanesyan ,&nbsp;Tatyana S. Stupina ,&nbsp;Natalia A. Sanina","doi":"10.1016/j.jinorgbio.2024.112747","DOIUrl":"10.1016/j.jinorgbio.2024.112747","url":null,"abstract":"<div><div>A new promising binuclear tetranitrosyl iron complex with 2-methoxythiophenolyl of the composition [Fe₂(C₇H₇OS)₂(NO)₄] (complex <strong>1</strong>), which acts on the therapeutic targets of cardiovascular diseases, guanylate and adenylate cyclase, has been synthesized. X-ray diffraction data show the presence of two isoforms of complex <strong>1</strong>; according to quantum chemical calculations, the structure of only the <em>trans</em> isomer is stable in solutions.</div><div>The processes of transformation of complex <strong>1</strong> in DMSO, in aqueous solutions, as well as in the presence of bovine serum albumin, reduced glutathione, and mucin were studied. DMSO promotes the decomposition of the original complex <strong>1</strong> into mononuclear products. In biological systems, the mechanisms of decomposition of the complex <strong>1</strong> differ from aqueous solutions. In albumin solution, a gradual formation of a high-molecular-weight dinitrosyl complex is observed, obtained by coordinating the [Fe(NO)₂]⁺ fragment with the amino acid groups of the protein. In the presence of mucin, an EPR signal from stable mononitrosyl products is observed. The introduction of glutathione into the system of the complex <strong>1</strong> leads to the replacement of one initial thioligand with glutathione. In the model systems under study, a more efficient and prolonged generation of NO groups is observed compared to a buffer solution.</div><div>The obtained data on the influence of the environment on the properties of the complex <strong>1</strong> in combination with a study of their effect on enzymes allow us to recommend it for further study as a potential drug with vasodilator, antianginal, and hypotensive properties.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142374877","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":"Uridine triphosphate hybrid catalyst for carbon‑carbon bond formation reactions with enhanced enantioselectivity by mercury(II) ions","authors":"Li Liu,&nbsp;Xingchen Dong,&nbsp;Weijun Qin,&nbsp;Yashao Chen,&nbsp;Changhao Wang","doi":"10.1016/j.jinorgbio.2024.112748","DOIUrl":"10.1016/j.jinorgbio.2024.112748","url":null,"abstract":"<div><div>DNA hybrid catalysts are constructed by embedding active metal species into the chiral scaffolds of DNA, which have been successfully applied to some important aqueous-phase enantioselective transformations. Owing to simple components and inherent chirality, nucleotide hybrid catalysts are emerging in response to soving the unclear locations of catalytic centers and the plausible catalytic mechanisms in DNA-based asymmetric catalysis. However, the tertiary structure of nucleotides lacks tunability, severely impeding further design of nucleotide hybrid catalysts for potential applications. To this end, a design strategy for tunable nucleotide hybrid catalysts is put forward by introducing metal-mediated base pairs. Herein, we found that the formation of uracil‑mercury(II)-uracil (U-Hg²⁺-U) base pairs could enhance the enantioselectivity in uracil-containing nucleotide-based asymmetric reactions. Compared with uracil triphosphate (UTP) complexing with Cu²⁺ ions (UTP∙Cu²⁺), the presence of Hg²⁺ ions gave rise to an increased enantiomeric excess (ee) of 38 % in Diels-Alder reactions and 22 % ee in Michael reactions. The Hg²⁺-tuning behaviors of UTP hybrid catalyst have been demonstrated to largely depend on nucleotides, Hg²⁺ concentrations, metal cofactors, additives and reaction types. Based on ultraviolet-visible, circular dichroism and nuclear magnetic resonance spectroscopic techniques, the chiral enhancement of Hg²⁺-containing UTP hybrid catalyst is proved to largely depend on the formation of U-Hg²⁺-U base pairs and the plausible cross-linked structure of UTP-Hg²⁺-UTP/Cu²⁺ assembly. This work provides a tunable strategy based on the concept of metal-mediated base pairs, allowing further design of potent oligonucleotide-based catalysts for other enantioselective reactions.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142370597","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":"Pd(II)/1,10-phenanthroline complexes bearing arene ligands: On the role of N- vs O-coordination to tune their cellular activity and binding ability towards DNA and RNA","authors":"Francesca Binacchi ,&nbsp;Damiano Cirri ,&nbsp;Eleonora Bimbi ,&nbsp;Natalia Busto ,&nbsp;Alessandro Pratesi ,&nbsp;Tarita Biver","doi":"10.1016/j.jinorgbio.2024.112749","DOIUrl":"10.1016/j.jinorgbio.2024.112749","url":null,"abstract":"<div><div>Three Pd(II)-based complexes of 1,10-phenanthroline and <em>N</em>- or <em>O</em>-coordinating ligands have been synthesised and tested with different relevant biosubstrates like double-stranded DNA, double and triple helix of RNA, DNA G-quadruplexes of different conformations and bovine serum albumin. Here a correlation between N- vs O-coordinating elements and binding mechanism emerged, where the N-coordinating ligands proved to be the most promising. These outcomes were confirmed also in the cellular experiments. The Pd(II) complex with naphthalene-1,8-diamine is the one that is able to be carried by BSA, to strongly bind nucleic acids, to produce reactive oxygen species (ROS) and to show the best cellular performances (poorly toxic towards healthy cells and highly toxic against the cisplatin-resistant cancer cell line). On the opposite, the complex with benzene-1,2-diolate may be sequestered by BSA, weakly binds nucleic acids, does not produce ROS and shows poor cellular activity. The complex with benzene-1,2-diamine stays in between. Other mechanistic details are discussed which show that the biophysical behaviour is the sum of the contribution of aromaticity, charge and <em>N-</em> or <em>O-</em>coordination.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142374876","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to \"New insights into the O₂-sensing mechanism of FixL and other gas sensing heme proteins\" [Journal of Inorganic Biochemistry 259 (2024) 112642].","authors":"Mark F Reynolds","doi":"10.1016/j.jinorgbio.2024.112746","DOIUrl":"https://doi.org/10.1016/j.jinorgbio.2024.112746","url":null,"abstract":"","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338548","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":"Generating globin-like reactivities in [human serum albumin-FeII(heme)] complex through N-donor ligand addition","authors":"Mary Grace I. Galinato ,&nbsp;Christopher Wyant ,&nbsp;Ashley L. Lombardo ,&nbsp;Ethan K. MacIsaac ,&nbsp;Daniella A. Rios-Martinez ,&nbsp;Christopher D. Kimrey ,&nbsp;Alexandra Alfonso Castro","doi":"10.1016/j.jinorgbio.2024.112743","DOIUrl":"10.1016/j.jinorgbio.2024.112743","url":null,"abstract":"<div><div>Human serum albumin (HSA) has a strong binding affinity for heme <em>b</em>, forming a complex in a 1:1 ratio with the co-factor ([HSA-Fe^IIIheme]). This system displays spectroscopic and functional properties comparable to globins when chemical derivatives mimicking them are incorporated into the protein matrix. The aim of this study is to generate globin-like systems using [HSA-Fe^IIIheme] as a protein template and binding N-donor ligands (imidazole, Im; and 1-methylimidazole, 1-MeIm) to construct artificial [HSA-Fe(heme)-(N-donor)] complexes. Their electronic structure and binding thermodynamics are investigated using UV–vis and (synchronous) fluorescence spectroscopies, while ligand-protein interactions are visualized using docking simulations. The imidazole derivatives have a strong affinity for [HSA-Fe^IIIheme] (K ∼ 10⁴–10⁶), where the spontaneous binding of Im and 1-MeIm are dominated by entropic and enthalpic effects, respectively. The reduced form of the [HSA-Fe(heme)-(N-donor)] complexes demonstrate nitrite reductase (NiR) activity similar to that observed in globins, but with significant differences in their rates. [HSA-Fe^IIheme-(1-MeIm)] reduces nitrite ∼4× faster than the Im analogue, and ∼ 30× faster than myoglobin (Mb). The enhanced NiR activity of [HSA-Fe^IIheme-(1-MeIm)] is a cumulative effect of several factors including a slightly expanded and more optimal heme binding pocket, nearby residues as possible proton sources, and a H-bonding interaction between 1-MeIm and residues Arg160 and Lys181 that may have a long-distance influence on the heme π electron density.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142359426","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":"The application of X-ray fluorescence microscopy and micro-XANES spectroscopy to study neuro-metallomics.","authors":"Meg Willans, Ashley Hollings, Rhiannon E Boseley, Thomas Munyard, Gaewyn C Ellison, Mark J Hackett","doi":"10.1016/j.jinorgbio.2024.112744","DOIUrl":"https://doi.org/10.1016/j.jinorgbio.2024.112744","url":null,"abstract":"<p><p>This early career research highlight provides a review of my own research program over the last decade, a time frame that encompasses my transition from postdoctoral fellowships to independent researcher. As an analytical chemist and applied spectroscopist, the central theme of my research program over this time has been protocol development at synchrotron facilities, with the main objective to investigate brain metal homeostasis during both brain health and brain disease. I will begin my review with an overview of brain metal homeostasis, before introducing analytical challenges associated with its study. I will then provide a brief summary of the two main X-ray techniques I have used to study brain metal homeostasis, X-ray fluorescence microscopy (XFM) and X-ray absorption near edge structure spectroscopy (XANES). The review then finishes with a summary of my main research contributions using these two techniques, put in the context of the results from others in the field.</p>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142338549","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":"Expanding the scope of resonance Raman spectroscopy in hydrogenase research: New observable states and reporter vibrations","authors":"Cornelius C.M. Bernitzky ,&nbsp;Giorgio Caserta ,&nbsp;Stefan Frielingsdorf ,&nbsp;Janna Schoknecht ,&nbsp;Andrea Schmidt ,&nbsp;Patrick Scheerer ,&nbsp;Oliver Lenz ,&nbsp;Peter Hildebrandt ,&nbsp;Christian Lorent ,&nbsp;Ingo Zebger ,&nbsp;Marius Horch","doi":"10.1016/j.jinorgbio.2024.112741","DOIUrl":"10.1016/j.jinorgbio.2024.112741","url":null,"abstract":"<div><div>Oxygen-tolerant [NiFe] hydrogenases are valuable blueprints for the activation and evolution of molecular hydrogen under application-relevant conditions. Vibrational spectroscopic techniques play a key role in the investigation of these metalloenzymes. For instance, resonance Raman spectroscopy has been introduced as a site-selective approach for probing metal-ligand coordinates of the [NiFe] active site and FeS clusters. Despite its success, this approach is still challenged by a limited number of detectable active-site states – due to missing resonance enhancement or intrinsic light sensitivity – and difficulties in their assignment. Utilizing two oxygen-tolerant [NiFe] hydrogenases as model systems, we illustrate how these challenges can be met by extending excitation and detection wavelength regimes in resonance Raman spectroscopic studies. Specifically, we observe that this technique does not only probe low-frequency metal-ligand vibrations but also high-frequency intra-ligand modes of the diatomic CO/CN⁻ ligands at the active site of [NiFe] hydrogenases. These reporter vibrations are routinely probed by infrared absorption spectroscopy, so that direct comparison of spectra from both techniques allows an unambiguous assignment of states detected by resonance Raman spectroscopy. Moreover, we find that a previously undetected state featuring a bridging hydroxo ligand between Ni and Fe can be probed using higher excitation wavelengths, as photoconversion occurring at lower wavelengths is avoided. In summary, this study expands the applicability of resonance Raman spectroscopy to hydrogenases and other complex metalloenzymes by introducing new strategies for probing and assigning redox-structural states of the active site.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142320357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"EcNikA, a versatile tool in the field of artificial metalloenzymes","authors":"Caroline Marchi-Delapierre,&nbsp;Christine Cavazza,&nbsp;Stéphane Ménage","doi":"10.1016/j.jinorgbio.2024.112740","DOIUrl":"10.1016/j.jinorgbio.2024.112740","url":null,"abstract":"<div><div>This review describes the multiple advantages of using of <em>Ec</em>NikA, a nickel transport protein, in the design of artificial metalloenzymes as alternative catalysts for synthetic biology. The rationale behind the strategy of artificial enzyme design is discussed, with particular emphasis on <em>de novo</em> active site reconstitution. The impact of the protein scaffold on the artificial active site and thus the final catalytic properties is detailed, highlighting the considerable aptitude of hybrid systems to catalyze selective reactions, from alkene to thioether transformations (epoxidation, hydroxychlorination, sulfoxidation). The different catalytic approaches – from <em>in vitro</em> to <em>in cristallo</em> – are compared, revealing the considerable advantages of protein crystals in terms of stabilization and acceleration of reaction kinetics. The versatility of proteins, based on metal and ligand diversity and medium/physical conditions, are thus illustrated for oxidation catalysis.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142454989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis, characterization and comparative biological activity of a novel set of Cu(II) complexes containing azole-based ligand frames","authors":"Courtney E. Elwell ,&nbsp;Emily Stein ,&nbsp;Adam Lewis ,&nbsp;Stefan Hamaway ,&nbsp;Kennedy A. Alexis ,&nbsp;Joseph M. Tanski ,&nbsp;Timothy J. Barnum ,&nbsp;Colleen M. Connelly ,&nbsp;Laurie A. Tyler","doi":"10.1016/j.jinorgbio.2024.112736","DOIUrl":"10.1016/j.jinorgbio.2024.112736","url":null,"abstract":"<div><div>The synthesis and spectroscopic characterization of three complexes containing a substituted 2-(2-pyridyl)benzothiazole (PyBTh) group in the ligand frame are reported along with the comparative biological activity. The ligands have been substituted at the 6-position with either a methoxy (Py(<em>OMe</em>)BTh) or a methyl group (Py(<em>Me</em>)BTh). Reaction of Py(<em>OMe</em>)BTh with either CuCl₂ or Cu(NO₃)₂·2.5 H₂O yielded the monomeric [Cu(Py(<em>OMe</em>)BTh))₂(NO₃)]NO₃·1.5 MeOH, (<strong>1</strong>·1.5 MeOH) complex or the dimeric [Cu(Py(<em>OMe</em>)BTh)Cl₂]₂ (<strong>2</strong>), respectively, with the nuclearity of the complex dependent on the starting Cu(II) salt. Reaction between the methyl substituted ligand and Cu(NO₃)₂·2.5 H₂O resulted in the isolation of Cu(Py(<em>Me</em>)BTh)(NO₃)₂·0.5 THF (<strong>3</strong>·0.5 THF). Complexes <strong>1</strong>–<strong>3</strong> were fully characterized. Cyclic voltammetry measurements were performed on all three complexes as well as on [Cu(PyBTh)₂(H₂O)](BF₄)₂ (<strong>4</strong>), a compound previously reported by us which contains the unsubstituted 2-(2-pyridyl)benzothiazole ligand. The biological activity was studied and included concentration dependent DNA binding and cleavage, antibacterial activity, and cancer cell toxicity. All complexes exhibited DNA cleavage activity, however <strong>2</strong> and <strong>4</strong> were found to be the most potent. Mechanistic studies revealed that the nuclease activity is dependent on an oxidative mechanism reliant principally on O₂⁻. Antibacterial studies revealed complex <strong>4</strong> was more potent compared to <strong>1</strong>–<strong>3</strong>. Cancer cell toxicity studies were carried out on HeLa, PC-3, and MCF7 cells with <strong>1</strong>–<strong>4</strong>, Cu(QBTh)(NO₃)₂(H₂O) and Cu(PyBIm)₃(BF₄)₂. The differences in the observed toxicities suggests the importance of the ligand and its substituents in modulating cell death.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142322293","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":"Mononuclear high-spin iron(III) phthalocyanines","authors":"Yusuke Okada,&nbsp;Nagao Kobayashi","doi":"10.1016/j.jinorgbio.2024.112737","DOIUrl":"10.1016/j.jinorgbio.2024.112737","url":null,"abstract":"<div><div>2,9(or 10),16(or 17), 23(or 24)-Tetradecyloxycarbonylphthalocyaninatoiron, FeTDPc, and 2,3,9,10,16,17,23,24-octadecyloxycarbonylphthalocyaninatoiron, FeODPc, were synthesized and characterized. These compounds seem to be in trivalent iron high-spin state in solvents such as chloroform, dichloromethane, benzene, and chlorobenzene, although their counter anion could not be detected by elemental analyses. They react with strong bases such as pyridine and imidazoles to form their mono- and subsequently their di-base complexes with formation constant of &gt;10⁶ and &lt; 200 dm³ mol⁻¹, respectively, in dichloromethane at 20 °C. The resultant mono-adducts appear to be trivalent iron low-spin while the di-base adducts are bivalent iron low-spin state complexes. The addition of ca. 10–30 equivalent of tetrabutylammonium-chloride or -bromide (electrolyte) to the solution containing FeTDPc or FeODPc, causes their spin-state change from iron(III) high to low-spin state. In a solid power state, however, both FeTDPc and FeODPc exist as a mixture of high-spin iron(III)- and intermediate-spin iron(II) species. Strangely, when these compounds are dissolved in polystyrene, i.e. each molecules are isolated from each other, the signals originated from the iron(II) component disappear.</div></div>","PeriodicalId":364,"journal":{"name":"Journal of Inorganic Biochemistry","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142314594","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}