bioRxiv - Bioengineering最新文献

{"title":"Supramolecular Enzyme-Peptide Gels for Localized Therapeutic Biocatalysis","authors":"Madeline Fuchs, Dillon T Seroski, Bethsymarie Soto Morales, Lucas Melgar, Giannia Scibilio, Abigail Ziegler, Renjie Liu, Benjamin G Keselowsky, Gregory A Hudalla","doi":"10.1101/2024.09.06.611628","DOIUrl":"https://doi.org/10.1101/2024.09.06.611628","url":null,"abstract":"Enzyme therapeutics are often compromised by poor accumulation within target tissues, necessitating high doses that can exacerbate off-target effects. We report an injectable supramolecular enzyme-peptide gel platform for prolonged local enzyme retention in vivo. The gel is based on CATCH(+/-) (Co-Assembling Tags based on CHarge-complementarity), cationic and anionic peptide pairs that form β-sheet fibrils upon mixing. Enzymes recombinantly fused to CATCH(-) peptide integrate into CATCH(+/-) β-sheet fibrils during assembly, resulting in an enzyme-peptide gel. Catalytically-active gels were fabricated with four disparate enzymes: CATCH(-)-NanoLuc luciferase, CATCH(-)-cutinase, CATCH(-)-uricase, and CATCH(-)-adenosine synthase A. CATCH(-)-cutinase gels demonstrated tunability of the platform, while CATCH(-)-NanoLuc gels demonstrated prolonged tissue retention relative to soluble enzyme. CATCH(-)-uricase gels resolved localized inflammation in a gout model, while CATCH(-)-adenosine synthase A gels suppressed localized lipopolysaccharide-induced inflammation. Modular and tunable enzyme content, coupled with prolonged tissue retention, establish CATCH enzyme-peptide gels as a generalizable vehicle for effective local therapeutic biocatalysis.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"39 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Surface-Based vs. Voxel-Based Finite Element Head Models: Comparative Analyses of Strain Responses","authors":"Zhou Zhou, Xiaogai Li, Svein Kleiven","doi":"10.1101/2024.09.04.611159","DOIUrl":"https://doi.org/10.1101/2024.09.04.611159","url":null,"abstract":"Finite element (FE) models of the human head are important injury assessment tools but developing a high-quality, hexahedral-meshed FE head model without compromising geometric accuracy is a challenging task. Important brain features, such as the cortical folds and ventricles, were captured only in a handful of FE head models that were primarily developed from two meshing techniques, i.e., surface-based meshing with conforming elements to capture the interfacial boundaries and voxel-based meshing by converting the segmented voxels into elements with and without meshing smoothing. Despite these advancements, little knowledge existed of how similar the strain responses were between surface- and voxel-based FE head models. To address this, a previously developed surface-based head model with conforming meshes to capture the cortical folds-subarachnoid cerebrospinal fluid and brain-ventricle interfaces was reused, and two voxel-based models with and without mesh smoothing were newly created here. These three models were employed to simulate head impacts. The results showed remarkable similarities in the strain responses between the surface- and the voxel-based models. When calculating commonly used injury metrics, including the percentile strains below the maximum (e.g., 95 percentile strain) and the volume of brain element with the strain over certain thresholds, the responses of the three models were virtually identical. When examining the strain distribution, the three models showed different patterns at the interfacial boundary (e.g., sulci and gyri in the cortex, regions adjacent to the falx and tentorium) with strain differences exceeding 0.1. The mesh smoothing procedure in the voxel-based models marginally reduced the strain discrepancies compared to the surface-based model. This study yielded new quantitative insights into the general similarity in the strain responses between the surface- and voxel-based FE head models and underscored that caution should be exercised when using the strain at the interface to predict injury.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"477 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216639","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Machine Learning Driven Optimization for High Precision Cellular Droplet Bioprinting","authors":"Jaemyung Shin, Minseok Kang, Kinam Hyun, Zhangkang Li, Hitendra Kumar, Kangsoo Kim, Simon S. Park, Keekyoung Kim","doi":"10.1101/2024.09.04.611131","DOIUrl":"https://doi.org/10.1101/2024.09.04.611131","url":null,"abstract":"Controlled volume microliter cell-laden droplet bioprinting is important for precise biologics deposition, reliably replicating 3D microtissue environments for building cell aggregates or organoids. To achieve this, we propose an innovative machine-learning approach to predict cell-laden droplet volumes according to input parameters. We developed a novel bioprinting platform capable of collecting high-throughput droplet images and generating an extensive dataset for training machine learning and deep learning algorithms. Our research compared the performance of three machine learning and two deep learning algorithms that predict droplet volume based on numerous bioprinting parameters. By adjusting bioink viscosity, nozzle size, printing time, printing pressure, and cell concentration as input parameters, we precisely could control droplet sizes, ranging from 0.1 to 50 microliter in volume. We utilized a hydrogel precursor composed of 5% gelatin methacrylate and a mixture of 0.5% and 1% alginate, respectively. Additionally, we optimized the cell bioprinting process using green fluorescent protein-tagged 3T3 fibroblast cells. These models demonstrated superior predictive accuracy and revealed the interrelationships among parameters while taking minimal time for training and testing. This method promises to advance the mass production of organoids and microtissues with precise volume control for various biomedical applications.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"53 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216618","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systemic in utero gene editing as a treatment for cystic fibrosis","authors":"Adele S. Ricciardi, Christina Barone, Rachael Putman, Elias Quijano, Anisha Gupta, Richard Nguyen, Hanna Mandl, Alexandra S. Piotrowski-Daspit, Francesc Lopez-Giraldez, Valerie Luks, Mollie R. Freedman-Weiss, James S. Farrelly, Samantha Ahle, Peter M. Glazer, W. Mark Saltzman, David H. Stitelman, Marie E. Egan","doi":"10.1101/2024.09.04.611031","DOIUrl":"https://doi.org/10.1101/2024.09.04.611031","url":null,"abstract":"In utero gene editing has the potential to modify disease causing genes in multiple developing tissues before birth, possibly allowing for normal organ development, disease improvement, and conceivably, cure. In cystic fibrosis (CF), a disease that arises from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, there are signs of multiorgan disease affecting the function of the respiratory, gastrointestinal, and reproductive systems already present at birth. Thus, treating CF patients early is crucial for preventing or delaying irreversible organ damage. Here we demonstrate proof-of-concept of multiorgan mutation correction in CF using peptide nucleic acids (PNAs) encapsulated in polymeric nanoparticles and delivered systemically in utero. In utero editing was associated with sustained postnatal CFTR activity, at a level similar to that of wild-type mice, in both respiratory and gastrointestinal tissue, without detection of off-target mutations in partially homologous loci. This work suggests that systemic in utero gene editing represents a viable strategy for treating monogenic diseases before birth that impact multiple tissue types.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"283 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Predictive Modeling of Sleep Slow Oscillation Emergence on the electrode manifold: Toward Personalized Closed-Loop Brain Stimulation","authors":"Mahmoud Alipour, Sara C. Mednick, Paola Malerba","doi":"10.1101/2024.09.03.611113","DOIUrl":"https://doi.org/10.1101/2024.09.03.611113","url":null,"abstract":"Background: Sleep slow oscillations (SOs), characteristic of NREM sleep, are causally tied to cognitive outcomes and the health-promoting homeostatic functions of sleep. Due to these known benefits, brain stimulation techniques aiming to enhance SOs are being developed, with great potential to contribute to clinical interventions, as they hold promise for improving sleep functions in populations with identified SO deficits (e.g., mild cognitive impairment). SO-targeting closed-loop stimulation protocols currently strive to identify SO occurrences in real time, a computationally intensive step that can lead to reduced precision (compared to post-hoc detection). These approaches are also often limited to focusing on only one electrode location, thus inherently precluding targeting of SOs that is informed by the overall organization of SOs in space-time. Prediction of SO emergence across the electrode manifold would establish an alternative to online detection, thus greatly advancing the development of personalized and flexible brain stimulation paradigms. This study presents a computational model that predicts SO occurrences at multiple locations across a night of sleep. In combination with our previous study on optimizing brain stimulation protocols using the spatiotemporal properties of SOs, this model contributes to increasing the accuracy of SO targeting in brain stimulation applications.\u0000Methods: SOs were detected in a dataset of nighttime sleep of 22 subjects (9 females), acquired with polysomnography including 64 EEG channels. Modeling of SO occurrence was achieved for SOs in stage N3, or in a combination of stages N2 and N3 (N2&N3). We study SO emergence at progressively more refined time scales. First, the cumulative SO occurrences in successive sleep cycles were successfully fit with exponentials. Secondly, the SO timing in each individual was modeled with a renewal point process. Using an inverse Gaussian model, we estimated the probability density function of SO timing and its parameters μ (mean) and λ (shape, representing skewness) in successive cycles.\u0000Results: We observed a declining trend in the SO count across sleep cycles, which we modeled using a power law relationship. The decay rate per cycle was 1.473 for N3 and 1.139 for N2&N3, with variances of the decay rates across participants being 1 and 0.53, respectively. This pattern mirrors the declining trend of slow wave activity (SWA) across sleep cycles, likely due to the inherent relationship between SWA and SO. Additionally, the SO timing model for N3 showed an increasing trend in the model parameters (μ, λ) across cycles. The increase rate per cycle followed a power law relationship with a rate of 0.83 and an exponential relationship with a rate of 4.59, respectively. The variances of the increase rates were 0.02 for μ and 0.44 for λ across participants.\u0000Conclusion: This study establishes a predictive model for SO occurrence during NREM sleep, providing insights into i","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modular photostable fluorescent DNA blocks dissect the effects of pathogenic mutant kinesin on collective transport","authors":"Tomoki Kita, Ryota Sugie, Yuki Suzuki, Shinsuke Niwa","doi":"10.1101/2024.09.06.611758","DOIUrl":"https://doi.org/10.1101/2024.09.06.611758","url":null,"abstract":"Intracellular transport is driven by teams of various motor proteins. Advances in DNA nanotechnology have enabled the programmable linkage of different types of motor proteins. In this study, we developed a modular, photostable, fluorescence-labeled tiny DNA origami block (FTOB) for extended observation of collective transport by selected motor combinations. The FTOB is designed as a 4-helix bundle (~8.4 nm) with densely accumulated fluorescent dyes, minimizing blinking and photobleaching. By designing a pair of connector DNAs, FTOBs are heterodimerized following motor protein attachment using the ALFA-tag/nanobody system. Our system examined the impact of a pathogenic mutant kinesin on its collective movement with wild-type kinesin, clearly observing two distinct behaviors: the team's velocity was generally governed by the slower mutant but occasionally surged to levels comparable to that of a single wild-type motor. Our photostable, robust, modular FTOB system could serve as a versatile tool for precisely dissecting cooperative cargo transport.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"19 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Digital Microfluidic Platform for the Microscale Production of Functional Immune Cell Therapies","authors":"Samuel R. Little, Niloufar Rahbari, Mehri Hajiaghayi, Fatemeh Gholizadeh, Fanny-Mei Cloarec-Ung, Joel Phillips, Hugo Sinha, Alison Hirukawa, David JHF Knapp, Peter J. Darlington, Steve CC Shih","doi":"10.1101/2024.09.03.611092","DOIUrl":"https://doi.org/10.1101/2024.09.03.611092","url":null,"abstract":"Genetically engineering human immune cells has been shown to be an effective approach for developing novel cellular therapies to treat a wide range of diseases. To expand the scope of these cellular therapies while solving persistent challenges, extensive research and development is still required. Electroporation has recently emerged as one of the most popular techniques for inserting biological payloads into human immune cells to perform genetic engineering. However, several recent studies have reported that electroporation can negatively impact cell functionality. Additionally, the requirement to use large amounts of cells and expensive payloads to achieve efficient delivery can drive up the costs of development efforts. Here we use a digital microfluidic enabled electroporation system (referred to as triDrop) and compare them against two state-of-the-art commercially available systems for the engineering of human T cells. We describe the ability to use triDrop for highly viable, highly efficient transfection while using substantially fewer cells and payload. Subsequently, we perform transcriptomic analysis on cells engineered with each of the three systems and show that electroporation with triDrop lead to less dysregulation of several functionally relevant pathways. Finally, in a direct comparison of immunotherapeutic functionality, we show that T cells engineered with triDrop have an improved ability to mount an immune response when presented with tumor cells. These results show that the triDrop platform is uniquely suited to produce functionally engineered immune cells while also reducing the costs of cell engineering compared to other commercially available systems.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216641","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Computational Framework to Optimize the Mechanical Behavior of Synthetic Vascular Grafts","authors":"David Jiang, Andrew J Robinson, Abbey Nkansah, Jonathan Leung, Leopold Guo, Steve A Maas, Jeffrey A Weiss, Elizabeth M Cosgriff-Hernandez, Lucas H Timmins","doi":"10.1101/2024.09.05.608688","DOIUrl":"https://doi.org/10.1101/2024.09.05.608688","url":null,"abstract":"The failure of synthetic small-diameter vascular grafts has been attributed to a mismatch in the compliance between the graft and native artery, driving mechanisms that promote thrombosis and neointimal hyperplasia. Additionally, the buckling of grafts results in large deformations that can lead to device failure. Although design features can be added to lessen the buckling potential, the addition is detrimental to decreasing compliance (e.g., reinforcing coil). Herein, we developed a novel finite element framework to inform vascular graft design by evaluating compliance and resistance to buckling. A batch-processing scheme iterated across the multi-dimensional design parameter space, which included three parameters: coil thickness, modulus, and spacing. Three types of finite element models were created in FEBio for each unique coil-reinforced graft parameter combination to simulate pressurization, axial buckling, and bent buckling, and results were analyzed to quantify compliance, buckling load, and kink radius, respectively, from each model. Importantly, model validation demonstrated that model predictions agree qualitatively and quantitatively with experimental observations. Subsequently, data for each design parameter combination were integrated into an optimization function for which a minimum value was sought. The optimization values identified various candidate graft designs with promising mechanical properties. Our investigation successfully demonstrated the model-directed framework identified vascular graft designs with optimal mechanical properties, which can potentially improve clinical outcomes by addressing device failure. In addition, the presented computational framework promotes model-directed device design for a broad range of biomaterial and regenerative medicine strategies.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"181 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216622","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating Joint Angle Data for Clinical Assessment Using Multidimensional Inverse Kinematics with Average Segment Morphometry.","authors":"Rachel I Taitano, Valeriya Gritsenko","doi":"10.1101/2024.09.03.611088","DOIUrl":"https://doi.org/10.1101/2024.09.03.611088","url":null,"abstract":"Movement analysis is a critical tool in understanding and addressing various disabilities associated with movement deficits. By analyzing movement patterns, healthcare professionals can identify the root causes of these alterations, which is essential for preventing, diagnosing, and rehabilitating a broad spectrum of medical conditions, disabilities, and injuries. With the advent of affordable motion capture technologies, quantitative data on patient movement is more accessible to clinicians, enhancing the quality of care. Nonetheless, it is crucial that these technologies undergo rigorous validation to ensure their accuracy in collecting and monitoring patient movements, particularly for remote healthcare services where direct patient observation is not possible. In this study, motion capture technology was used to track upper extremity movements during a reaching task presented in virtual reality. Kinematic data was then calculated for each participant using a scaled dynamic inertial model. The goal was to evaluate the accuracy of joint angle calculations using inverse kinematics from motion capture relative to the typical movement redundancy. Shoulder, elbow, radioulnar, and wrist joint angles were calculated with models scaled using either direct measurements of each individual’s arm segment lengths or those lengths were calculated from individual height using published average proportions. The errors in joint angle trajectories calculated using the two methods of model scaling were compared to the inter-trial variability of those trajectories. The variance of this error was primarily within the normal range of variability between repetitions of the same movements. This suggests that arm joint angles can be inferred with good enough accuracy from motion capture data and individual height to be useful for the clinical assessment of motor deficits.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"36 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216646","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of Acetate and Optimized Nitrate Levels on Mixotrophic Growth and Protein Dynamics in Chlorella Sorokiniana","authors":"Sunni Chen, Ruiqi Wang, Youn Joong Kim, Emily Radican, Yu Lei, Yong Ku Cho, Zhenlei Xiao, Mingyu Qiao, Yangchao Luo","doi":"10.1101/2024.09.04.611160","DOIUrl":"https://doi.org/10.1101/2024.09.04.611160","url":null,"abstract":"Microalgae are well-known for their role as sustainable bio-factories, offering a promising solution to the global food and nutrition crisis. To clarify the potential of Chlorella sorokiniana UTEX 1230 for food applications, particularly as an alternative protein source, the study employed a mixotrophic cultivation mode with sodium acetate (NaAc) as a cost-effective organic carbon (NaAc-C) source. Varying levels of NaAc-C and nitrate-sourced nitrogen were investigated, optimizing the effect of metabolic characteristics of the microalgal growth. The designed heterotrophic cultivation confirmed the ability of C. sorokiniana UTEX 1230 to grow on NaAc-C, and then the mixotrophic cultures, when supported by both NaAc-C and CO2, exhibited superior growth performance, achieving double the biomass concentration compared to the autotrophic control. The addition of nitrogen (750 mg/L NaNO₃) facilitated the thorough metabolism of NaAc-C and enhanced photosynthetic activity indicated by a 196% increase in pigment levels, which resulted in a maximum biomass concentration of 2.82 g/L in the 150 mM NaAc-C group. A detailed analysis of nitrogen and protein concentrations over time revealed that higher nitrogen availability led to greater protein accumulation which was then degraded to support essential life activities under nitrogen starvation. Therefore, it is suggested that supplementing nitrate on the 3rd day and harvesting on the 4th day could be strategically implemented to increase protein yield from 0.17 g/L/d to 0.34 g/L/d. These findings offer theoretical guidance for further refining this microalgal strain for use as an alternative protein.","PeriodicalId":501308,"journal":{"name":"bioRxiv - Bioengineering","volume":"3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142216640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}