{"title":"Genetically Engineered Liposwitch-Based Nanomaterials.","authors":"Md Shahadat Hossain, Alex Wang, Salma Anika, Zhe Zhang, Davoud Mozhdehi","doi":"10.1021/acs.biomac.4c01388","DOIUrl":"10.1021/acs.biomac.4c01388","url":null,"abstract":"<p><p>Fusion of intrinsically disordered and globular proteins is a powerful strategy to create functional nanomaterials. However, the immutable nature of genetic encoding restricts the dynamic adaptability of nanostructures postexpression. To address this, we envisioned using a myristoyl switch, a protein that combines allostery and post-translational modifications─two strategies that modify protein properties without altering their sequence─to regulate intrinsically disordered protein (IDP)-driven nanoassembly. A typical myristoyl switch, allosterically activated by a stimulus, reveals a sequestered lipid for membrane association. We hypothesize that this conditional exposure of lipids can regulate the assembly of fusion proteins, a concept we term \"liposwitching\". We tested this by fusing recoverin, a calcium-dependent myristoyl switch, with elastin-like polypeptide, a thermoresponsive model IDP. Biophysical analyses confirmed recoverin's myristoyl-switch functionality, while dynamic light scattering and cryo-transmission electron microscopy showed distinct calcium- and lipidation-dependent phase separation and assembly. This study highlights liposwitching as a viable strategy for controlling DP-driven nanoassembly, enabling applications in synthetic biology and cellular engineering.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"8058-8068"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632658/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142566661","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-08DOI: 10.1021/acs.biomac.4c00765
Andrea Malandrino, Huijun Zhang, Nico Schwarm, David Böhringer, Delf Kah, Christian Kuster, Aldo R Boccaccini, Ben Fabry
{"title":"Plasticity of 3D Hydrogels Predicts Cell Biological Behavior.","authors":"Andrea Malandrino, Huijun Zhang, Nico Schwarm, David Böhringer, Delf Kah, Christian Kuster, Aldo R Boccaccini, Ben Fabry","doi":"10.1021/acs.biomac.4c00765","DOIUrl":"10.1021/acs.biomac.4c00765","url":null,"abstract":"<p><p>Under 3D culture conditions, cells tend to spread, migrate, and proliferate better in more viscoelastic and plastic hydrogels. Here, we present evidence that the improved cell behavior is facilitated by the lower steric hindrance of a more viscoelastic and plastic matrix with weaker intermolecular bonds. To determine intermolecular bond stability, we slowly insert semispherical tipped needles (100-700 μm diameter) into alginate dialdehyde-gelatin hydrogels and measure stiffness, yield strength, plasticity, and the force at which the surface ruptures (puncture force). To tune these material properties without affecting matrix stiffness, we precross-link the hydrogels with CaCl<sub>2</sub> droplets prior to mixing in NIH/3T3 fibroblasts and final cross-linking with CaCl<sub>2</sub>. Precross-linking introduces microscopic weak spots in the hydrogel, increases plasticity, and decreases puncture force and yield strength. Fibroblasts spread and migrate better in precross-linked hydrogels, demonstrating that intermolecular bond stability is a critical determinant of cell behavior under 3D culture conditions.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7608-7618"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632650/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602394","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-25DOI: 10.1021/acs.biomac.4c01130
Yingzi Wang, Mingda Zhao, Yaping Zou, Xiaojuan Wang, Min Zhang, Yong Sun
{"title":"Hyaluronan Scaffold Decorated with Bifunctional Peptide Promotes Wound Healing via Antibacterial and Anti-Inflammatory.","authors":"Yingzi Wang, Mingda Zhao, Yaping Zou, Xiaojuan Wang, Min Zhang, Yong Sun","doi":"10.1021/acs.biomac.4c01130","DOIUrl":"10.1021/acs.biomac.4c01130","url":null,"abstract":"<p><p>The invasion of bacteria and inflammation impeded infected wounds heal. Here, a hyaluronan-based scaffold (HAG-<i>g</i>-C) was designed by cross-linking with gallic acid-modified gelatin to provide a protein microenvironment and decorated with cathelicidin-BF (CBF), a natural antimicrobial peptide, to remove bacterial infections and reverse the inflammatory environment. <i>In vitro</i>, HAG-<i>g</i>-C presented an antibacterial effect on <i>Staphylococcus aureus</i> and <i>Escherichia coli</i>. Meanwhile, it could drive the phenotypic switch of macrophage from M1 to M2 to accelerate tissue remodeling. In a mouse model of <i>S. aureus</i>-infected total skin defects, HAG-<i>g</i>-C inhibited the process of infection at the beginning of the wound and then regulated the M1 macrophage transformed to M2 phenotype on day 12. In addition, HAG-<i>g</i>-C induced collagen deposition, and reduced the expression of TNF-α, thereby significantly accelerating the reconstruction of infected wounds.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7850-7860"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142714784","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-18DOI: 10.1021/acs.biomac.4c01240
Belynn Sim, Jun Jie Chang, Qianyu Lin, Joey Hui Min Wong, Valerie Ow, Yihao Leow, Yi Jing Wong, Yi Jian Boo, Rubayn Goh, Xian Jun Loh
{"title":"Hydrogels Based on Polyelectrolyte Complexes: Underlying Principles and Biomedical Applications.","authors":"Belynn Sim, Jun Jie Chang, Qianyu Lin, Joey Hui Min Wong, Valerie Ow, Yihao Leow, Yi Jing Wong, Yi Jian Boo, Rubayn Goh, Xian Jun Loh","doi":"10.1021/acs.biomac.4c01240","DOIUrl":"10.1021/acs.biomac.4c01240","url":null,"abstract":"<p><p>Ionic complexes of electrostatically charged biomacromolecules are key players in various biological processes like nucleotide transportation, organelle formation, and protein folding. These complexes, abundant in biological systems, contribute to the function, responsiveness, and mechanical properties of organisms. Coherent with these natural phenomena, hydrogels formed through the complexation of oppositely charged polymers exhibit unique attributes, such as rapid self-assembly, hierarchical microstructures, tunable properties, and protective encapsulation. Consequently, polyelectrolyte complex (PEC) hydrogels have garnered considerable interest, emerging as an up-and-coming platform for various biomedical applications. This review outlines the underlying principles governing PEC hydrogels. The classification of polyelectrolytes and the self-assembly of PEC hydrogels are discussed, including the factors influencing their self-assembly process. Recent developments of PEC hydrogels for biomedical applications, including drug delivery, tissue engineering, wound healing and management, and wearable sensors, are summarized. This review concludes with the prospective directions for the next generation of PEC hydrogel research.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7563-7580"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142666416","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-07DOI: 10.1021/acs.biomac.4c01229
Leon Bixenmann, Taufiq Ahmad, Fabian Stephan, Lutz Nuhn
{"title":"End-Group Dye-Labeled Poly(hemiacetal ester) Block Copolymers: Enhancing Hydrolytic Stability and Loading Capacity for Micellar (Immuno-)Drug Delivery.","authors":"Leon Bixenmann, Taufiq Ahmad, Fabian Stephan, Lutz Nuhn","doi":"10.1021/acs.biomac.4c01229","DOIUrl":"10.1021/acs.biomac.4c01229","url":null,"abstract":"<p><p>Polymers with hemiacetal esters integrated in their backbone provide beneficial degradation profiles for (immuno-) drug delivery. However, their fast hydrolysis and low drug loading capacity have limited their applications so far. Therefore, this study focuses on the stability and loading capacity of hemiacetal ester polymers. The hydrophobicity of the micellar core has a tremendous effect on the hemiacetal ester stability. For that purpose, we introduce a new monomer with a phenyl moiety for stabilizing the micellar core and improving drug loading. The carrier functionality can further be expanded by post-polymerization modifications via activated ester groups at the polymer chain end. This allows for covalent dye labeling, which provides substantial insights into the polymers' <i>in vitro</i> performance. Flow cytometric analyses on RAW dual macrophages revealed intact micelles exhibiting significantly higher cellular uptake compared to degraded species, thus, highlighting the potential of end group functionalized poly(hemiacetal ester)s for (immuno)drug delivery purposes.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7958-7974"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602377","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-10-25DOI: 10.1021/acs.biomac.4c00883
Liliana B Hurtado, Mercedes Jiménez-Rosado, Maryam Nejati, Faiza Rasheed, Thomas Prade, Amparo Jiménez-Quero, Marcos A Sabino, Antonio J Capezza
{"title":"Genipap Oil as a Natural Cross-Linker for Biodegradable and Low-Ecotoxicity Porous Absorbents via Reactive Extrusion.","authors":"Liliana B Hurtado, Mercedes Jiménez-Rosado, Maryam Nejati, Faiza Rasheed, Thomas Prade, Amparo Jiménez-Quero, Marcos A Sabino, Antonio J Capezza","doi":"10.1021/acs.biomac.4c00883","DOIUrl":"10.1021/acs.biomac.4c00883","url":null,"abstract":"<p><p>Proteins derived from agroindustrial coproducts and a natural cross-linking agent (genipap oil containing genipin) were used to develop porous materials by reactive extrusion for replacing fossil-based absorbents. Incorporating genipap oil allowed the production of lightweight structures with high saline uptake (above 1000%) and competing retention capacity despite their porous nature. The mechanical response of the genipap-cross-linked materials was superior to that of the noncross-linked ones and comparable to those cross-linked using commercial genipin. The extruded products were hemocompatible and soil-biodegradable in less than 6 weeks. The compounds generated by the degradation process were not found to be toxic to the soil, showing a high bioassimilation capacity by promoting grass growth. The results demonstrate the potential of biopolymers and new green cross-linkers to produce fully renewable-based superabsorbents in hygiene products with low ecotoxicity. The study further promotes the production of these absorbents using low-cost proteins and continuous processing such as reactive extrusion.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7642-7659"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632661/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142491091","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-08DOI: 10.1021/acs.biomac.4c01011
Leila Tabrizi, Ross McGarry, Kaja Turzanska, Lazaros Varvarezos, Muireann Fallon, Ruairi Brannigan, John T Costello, Deirdre Fitzgerald-Hughes, Mary T Pryce
{"title":"Porphyrin-Polymer as a Photosensitizer Prodrug for Antimicrobial Photodynamic Therapy and Biomolecule Binding Ability.","authors":"Leila Tabrizi, Ross McGarry, Kaja Turzanska, Lazaros Varvarezos, Muireann Fallon, Ruairi Brannigan, John T Costello, Deirdre Fitzgerald-Hughes, Mary T Pryce","doi":"10.1021/acs.biomac.4c01011","DOIUrl":"10.1021/acs.biomac.4c01011","url":null,"abstract":"<p><p>This study presents the development and characterization of a novel porphyrin-Jeffamine polymer conjugate designed to function as a photosensitizer prodrug for antimicrobial photodynamic therapy (aPDT). The conjugate features a photosensitive porphyrin unit covalently attached to a biocompatible polymer backbone, with enhanced solubility, stability, and bioavailability compared to those of the free porphyrin derivatives. The photophysical properties were studied using transient absorption spectroscopy spanning the fs-μs time scales in addition to emission studies. The production of reactive oxygen species upon photoactivation enabled effective bacterial cell killing. Spectroscopic studies confirmed strong binding of the conjugate to DNA through intercalation, likely disrupting DNA replication and transcription. Interaction studies with bovine serum albumin demonstrated substantial serum protein binding, which may positively impact the pharmacokinetics and biodistribution. Overall, this porphyrin-polymer conjugate offers a multifunctional theranostic platform, combining antimicrobial action with DNA and protein binding potential, positioning it as a promising candidate for aPDT and bioimaging applications.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7736-7749"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632779/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142602434","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-11DOI: 10.1021/acs.biomac.4c01092
Antonín Edr, Dominika Wrobel, Alena Krupková, Lucie Červenková Št Astná, Evgeny Apartsin, Michaela Hympánová, Jan Marek, Jan Malý, Marek Malý, Tomáš Strašák
{"title":"Adaptive Synthesis, Supramolecular Behavior, and Biological Properties of Amphiphilic Carbosilane-Phosphonium Dendrons with Tunable Structure.","authors":"Antonín Edr, Dominika Wrobel, Alena Krupková, Lucie Červenková Št Astná, Evgeny Apartsin, Michaela Hympánová, Jan Marek, Jan Malý, Marek Malý, Tomáš Strašák","doi":"10.1021/acs.biomac.4c01092","DOIUrl":"10.1021/acs.biomac.4c01092","url":null,"abstract":"<p><p>Here, we present a modular synthesis as well as physicochemical and biological evaluation of a new series of amphiphilic dendrons carrying triphenylphosphonium groups at their periphery. Within the series, the size and mutual balance of lipophilic and hydrophilic domains are systematically varied, changing the dendron shape from cylindrical to conical. In physiological solution, the dendrons exhibit very low critical micelle concentrations (2.6-4.9 μM) and form stable and uniform micelles 6-12 nm in diameter, depending on dendron shape; the results correlate well with molecular dynamics simulations. The compounds show relatively high cytotoxicity (IC<sub>50</sub> 1.2-21.0 μM) associated with micelle formation and inversely related to the size of assembled particles. Depending on their shape, the dendrons show promising results in terms of dendriplex formation and antibacterial activity. In addition to simple amphiphilic dendrons, a fluorescently labeled analogue was also prepared and utilized as an additive visualizing the dendron's cellular uptake.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7799-7813"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11632778/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142612607","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-10-29DOI: 10.1021/acs.biomac.4c00828
Jane Yang, Madeline B Gelb, Kyle Tamshen, Neil L Forsythe, Jeong Hoon Ko, Ellie G Puente, Emma Pelegri-O'Day, Stephen M F Jamieson, Jo K Perry, Heather D Maynard
{"title":"Site-Selective Zwitterionic Poly(caprolactone-carboxybetaine)-Growth Hormone Receptor Antagonist Conjugate: Synthesis and Biological Evaluation.","authors":"Jane Yang, Madeline B Gelb, Kyle Tamshen, Neil L Forsythe, Jeong Hoon Ko, Ellie G Puente, Emma Pelegri-O'Day, Stephen M F Jamieson, Jo K Perry, Heather D Maynard","doi":"10.1021/acs.biomac.4c00828","DOIUrl":"10.1021/acs.biomac.4c00828","url":null,"abstract":"<p><p>Zwitterionic polymers have been found to be biocompatible alternatives to poly(ethylene glycol) (PEG) for conjugation to proteins. This work reports the site-selective conjugation of poly(caprolactone-carboxybetaine) (pCLZ) to human growth hormone receptor antagonist (GHA) B2036-alkyne and investigation of safety, activity, and pharmacokinetics. Azide-end-functionalized pCLZs were synthesized and conjugated to GHA B2036-alkyne via copper-catalyzed click reaction. The resulting inhibitory bioactivity concentration responses in Ba/F3-GHR cells were compared to those of PEGylated GHA B2036. IgG and IgM antibody production was tested in mice, and no measurable antibody or cytokine production was detected for the pCLZ conjugate. Using <sup>18</sup>F-labeled PET/CT imaging, the pCLZ conjugate showed an increase in circulation time compared to that of GHA B2036. Acute toxicity of the polymer was investigated in vivo and found to be nontoxic. Ex vivo degradation of the polymer on the conjugate was investigated. The results suggest that pCLZ-GHA is a potentially safe alternative to PEG-GHA.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7619-7629"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142542855","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}
BiomacromoleculesPub Date : 2024-12-09Epub Date: 2024-11-14DOI: 10.1021/acs.biomac.4c01203
Mónica C S Fernandes, Rita Branco, Patrícia Pereira, Jorge F J Coelho, Paula V Morais, Arménio C Serra
{"title":"Antimicrobial Activity of Copolymer Structures from Bio-Based Monomers.","authors":"Mónica C S Fernandes, Rita Branco, Patrícia Pereira, Jorge F J Coelho, Paula V Morais, Arménio C Serra","doi":"10.1021/acs.biomac.4c01203","DOIUrl":"10.1021/acs.biomac.4c01203","url":null,"abstract":"<p><p>The urgent need for new antimicrobial compounds has led scientists to explore antimicrobial peptides (AMPs) and antimicrobial polymers as solutions for multidrug resistance. In this study, we synthesized copolymers with cationic and hydrophobic moieties by free-radical polymerization (FRP) using a chain transfer agent to control molecular weights. The potential of natural products as part of the hydrophobic moiety was evaluated, along with variations in their monomer content (13-25%) and the molecular weight (MW) of the copolymer (5000-20,000 g·mol<sup>-1</sup>). Hydrophobicity was evaluated using the theoretical Log <i>P</i><sub>oct</sub> values and surface areas (SAs). Biological assays included antimicrobial activity against <i>Escherichia coli</i> and <i>Staphylococcus aureus</i> standard strains, hemolytic activity in red blood cells (RBC), and cytotoxicity tests against HEK293T cells. Keys findings indicate that copolymers with tropolone moieties, lower MWs, and an optimal balance between hydrophobic and cationic moieties show a promising basis for future generations of antimicrobials.</p>","PeriodicalId":30,"journal":{"name":"Biomacromolecules","volume":" ","pages":"7915-7925"},"PeriodicalIF":5.5,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142612610","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}