Biotechnology and applied biochemistry最新文献

{"title":"Acacetin Prevents Renal Damage Induced by Streptozotocin via Altering the NF-κB/ASC/NLRP3 and AMPK/SIRT1 Pathways in Mice.","authors":"Qingfei Yu, Hongyan Mao, Annamalai Vijayalakshmi, Meilan Zhou","doi":"10.1002/bab.2753","DOIUrl":"https://doi.org/10.1002/bab.2753","url":null,"abstract":"<p><p>Diabetic nephropathy (DN) is the most common cause of end-stage renal disease. Its pathogenesis includes inflammation, an excess of reactive oxygen species, and kidney damage. The present study intended to explore the nephroprotective effects of acacetin (ACN) in streptozotocin-induced diabetic animals. The following are the experimental groups: One millilitre of 0.9% saline was given to Group I (control), Streptozotocin (STZ) (diabetic animals) + 0.9% saline to Group II (DN group) (negative control), DN + ACN (15 mg/kg body weight [bw]) to Group III, and DN + Valsartan (150 mg/kg bw) to Group IV. According to the findings, ACN decreased the levels of glucose, serum creatinine (Scr), blood urea nitrogen (BUN), malondialdehyde (MDA), and proinflammatory cytokines while increasing the bw, superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in comparison to the DN animals. The histopathological analysis revealed that the animals treated with ACN showed recovery of renal damage in the tissues caused by STZ. In the STZ-induced DN mice, ACN reduced renal damage by upregulating the proteins of 5' adenosine monophosphate-activated protein kinase (AMPK), p-AMPK, and SIRT1 and downregulating the proteins of TGF-β, COL-1, COL-IV, NF-κB, ASC, NLRP3, and GSDMD, according to western blot analysis. Hence, the current study demonstrated that the regulation of the AMPK/SIRT1 and NF-κB/ASC/NLRP3 inflammasome pathways in DN mice was responsible for the protective effects of ACN. ACN may therefore be a viable treatment option for DN.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2753"},"PeriodicalIF":3.2,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143728718","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research Progress Fusion Tags for Recombinant Protein Production.","authors":"Jing-Jia Yuan, Shao-Lei Geng, Tian-Yun Wang","doi":"10.1002/bab.2749","DOIUrl":"https://doi.org/10.1002/bab.2749","url":null,"abstract":"<p><p>Recombinant proteins are obtained using genetic engineering techniques and are widely used in various fields. Some recombinant proteins are difficult to express, purify, or are unstable or insoluble due to their structural characteristics. In order to address such issues, additional tags are fused at either the N- or C-terminal end of the protein of interest during the cloning procedure. These tags range from a few residues to full-length proteins or domains not only maintaining the structure of the natural protein but can be used to improve the solubility, stability, yield, or to confer new properties of the target protein. Here, the fusion tags commonly used in recombinant protein production and their functions are reviewed, and novel fusion tags are also summarized.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2749"},"PeriodicalIF":3.2,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143728729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RETRACTION: Development of an Amperometric Biosensor That Can Determine the Amount of Glucose in the Blood Using the Glucose Oxidase Enzyme: Preparation of Polyaniline-Polypyrrole-Poly(sodium-4-styrenesulfonate) Film.","authors":"","doi":"10.1002/bab.2744","DOIUrl":"https://doi.org/10.1002/bab.2744","url":null,"abstract":"","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2744"},"PeriodicalIF":3.2,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143728793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DNA Ligase From Thermococcus radiotolerans: Recombinant Production, Characterization, Up-Scale Fermentation, and In Silico Evaluation.","authors":"Ahmed Osman, Saima Iftikhar, Bibi Nazia Murtaza, Salman Hosawi, Hisham N Tayeb, Thamir M Eid, Hayam A Alwabsi, Muhammad Shahid Nadeem","doi":"10.1002/bab.2747","DOIUrl":"https://doi.org/10.1002/bab.2747","url":null,"abstract":"<p><p>The present study describes molecular cloning, Escherichia coli expression, purification, and characterization of a DNA ligase from Thermococcus radiotolerans (TR). Optimal pH, temperature, media composition, and inducer concentrations for the production of wet cellular mass (WCM) with active enzyme have been validated. Recombinant DNA ligase-TR displayed a 44 kDa protein band on SDS-PAGE, and enzyme activity was measured at 50°C in the presence of 50 mM Tris-Cl buffer adjusted at pH 7 and supplemented with 0.4 mM ATP ± 0.1 mM NAD⁺. The enzyme was able to ligate two restriction products of EcoRI, independent of the presence of NAD⁺. The subject enzyme exhibited better activity than T4 DNA ligase. Maximum WCM was obtained at pH 7.3 and 30°C in modified M9NG medium when induced with 0.4 mM isopropyl β-d-1-thiogalactoside (IPTG). Protein sequence alignment studies have shown that DNA ligase-TR exhibits a novel and specific primary structure that is different from DNA ligases isolated from eukaryotic and other thermophilic species reported in the literature. According to in silico studies, Lys²³⁸ interacts with adenosine monophosphate (AMP) and the residues responsible to engage nicked DNA include Arg⁴¹, <math> <semantics><mrow><mi>Ar</mi> <msup><mi>g</mi> <mn>43</mn></msup> </mrow> <annotation>${mathrm{Ar}}{{mathrm{g}}^{43}}$</annotation></semantics> </math> , <math> <semantics><mrow><mi>Ar</mi> <msup><mi>g</mi> <mn>50</mn></msup> </mrow> <annotation>${mathrm{Ar}}{{mathrm{g}}^{50}}$</annotation></semantics> </math> , <math> <semantics><mrow><mi>Ar</mi> <msup><mi>g</mi> <mn>51</mn></msup> </mrow> <annotation>${mathrm{Ar}}{{mathrm{g}}^{51}}$</annotation></semantics> </math> , <math> <semantics><mrow><mi>Ar</mi> <msup><mi>g</mi> <mn>267</mn></msup> </mrow> <annotation>${mathrm{Ar}}{{mathrm{g}}^{267}}$</annotation></semantics> </math> , <math> <semantics><mrow><mi>Ar</mi> <msup><mi>g</mi> <mn>270</mn></msup> </mrow> <annotation>${mathrm{Ar}}{{mathrm{g}}^{270}}$</annotation></semantics> </math> , and Arg³⁶⁷. About 51% secondary structure consists of α-helices and β-pleated sheets. High-temperature stability, better half-life, and high activity of DNA ligase-TR advocate its suitability in the DNA manipulation procedures.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2747"},"PeriodicalIF":3.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143708692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Low-Fouling Electrochemical Immunosensor Based On Metal-Organic Framework and Ternary Metal Oxide Nanomaterials Using Dual Signal Amplification Strategy for Sensitive Detection of Prostate-Specific Antigen (PSA) in Patient Samples.","authors":"Abdulsalam Y Obaid, Zaman Abdalhussein Ibadi Alaridhee, Ahmed Hjazi, Nizomiddin Juraev, Marwah Suliman Maashi, Faeza H Zankanah, Ameer H Al-Rubaye, Albe Slabi Hussam, Mosleh Mohammad Abomughaid, Hasan Majdi, Majid Jabir","doi":"10.1002/bab.2752","DOIUrl":"https://doi.org/10.1002/bab.2752","url":null,"abstract":"<p><p>A label-free electrochemical immunosensor based on the zeolitic imidazolate framework-8 (ZIF8)/bismuth ferrite (BFO) nanocomposite was fabricated for the specific and sensitive quantification of prostate-specific antigen (PSA). The ZIF8-BFO material not only increases the surface area effectively but also enhances the catalytic capability of the electrode through a dual amplification strategy, leading to the improved sensitivity of the probe for PSA recognition. A thin layer of l-cysteine was used for two reasons: providing a scaffold for the next functionalization and reducing the fouling of plasma ingredients on the surface of the probe. The mechanical and spectroscopic properties of the produced nanomaterials were characterized using different techniques such as field emission scanning electron microscopy (FESEM), x-ray diffraction (XRD), atomic force microscopy (AFM), the Fourier transform infrared (FTIR), and dynamic light scattering (DLS)/Zeta analyzer. The electroanalytical properties of the probe were studied using square-wave voltammetry (SWV) and cyclic voltammetry (CV). The signal of the probe decreased proportionally with increasing PSA concentration in the 100.0 pg/mL-15.0 ng/mL range, with a limit of detection (LOD) of 85 pg/mL. The proposed platform has been successfully employed to measure PSA levels in human serum samples with acceptable accuracy. The capability of the probe was evaluated in detecting PSA in patient's serum samples, with results compared to those obtained from the gold standard enzyme-linked immunosorbent assay (ELISA). The results suggested that the ZIF8-BFO material-based probe could be used as a promising method for detecting PSA and tracing therapy progression in clinics.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2752"},"PeriodicalIF":3.2,"publicationDate":"2025-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143691273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Apelin-13 Inhibits the Adhesion of Monocytes to Endothelial Cells via the Gfi1/NF-κB Signaling Pathway.","authors":"Yujuan Wei, Zhihong Chen, Qi Zhao, Kaili Wang, Xiaojing Zhu, Guobing Zhou, Zhengke Yu","doi":"10.1002/bab.2746","DOIUrl":"https://doi.org/10.1002/bab.2746","url":null,"abstract":"<p><p>Oxidized low-density lipoprotein (ox-LDL)-induced endothelial dysregulation and the abnormal interaction between monocytes have been considered key risk factors for atherosclerosis. The study investigated the effects of apelin-13 on ox-LDL-induced endothelial dysfunction in human aortic endothelial cells (HAECs). Cells were stimulated with ox-LDL (100 mg/L), either alone or in combination with apelin-13 at concentrations of 3 and 6 µM. Multiple techniques, including real-time PCR, Western blot analysis, enzyme-linked immunosorbent assay (ELISA), cell attachment assays, and luciferase activity assays, were employed. Our results showed that ox-LDL reduced the expression of the G-protein-coupled apelin receptor (APJ) in HAECs. However, treatment with apelin-13 reduced the expression of lectin-like ox-LDL receptor 1 (LOX-1) against ox-LDL and inhibited the expression of pro-inflammatory cytokines in HAECs. Real-time PCR and ELISA assay demonstrated that apelin-13 also inhibited the expression of cell adhesion molecules intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin. The calcein AM staining method displayed that apelin-13 mitigated ox-LDL-induced attachment of THP-1 monocytes to HAECs. Furthermore, apelin-13 prevented the reduction of growth factor independence-1 (Gfi1) and the activation of NF-κB in HAECs, as evidenced by the luciferase activity assay. Knockdown of Gfi1 counteracted the inhibitory effects of apelin-13 on the attachment of THP-1 monocytes to HAECs, suggesting that the protective effects of apelin-13 in endothelial dysfunction are mediated by the Gfi1/ NF-κB signaling pathway. These findings suggest that apelin-13 may have therapeutic potential in preventing atherosclerosis by improving endothelial function and reducing inflammation.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2746"},"PeriodicalIF":3.2,"publicationDate":"2025-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143691276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of Invasive Weeds Lantana camara L. and Parthenium hysterophorus L. on the Secondary Metabolite Profiles of Okra (Abelmoschus esculentus L.).","authors":"Pratik Talukder, Mrinmoy Dasgupta, Aniket Hazra, Ayan Adhikari, Anushka Dutta, Snehika Sengupta, Milan Karmakar","doi":"10.1002/bab.2751","DOIUrl":"https://doi.org/10.1002/bab.2751","url":null,"abstract":"<p><p>The impact of two invasive weed species, Lantana camara and Parthenium hysterophorus, on the development and secondary metabolism of okra (Abelmoschus esculentus) is the focus of this study. We explored the potential role of secondary metabolites as antioxidants to mitigate the stress induced by these common weeds on okra plants with special emphasis on chlorogenic acid (CGA). The study has been done with six experimental setups. One setup includes okra grown alone, and another set consists of okra co-cultivation with L. camara, P. hysterophorus, and a combination of both. Each setup consisted of 30 plants of each species. After these, biochemical assays were done to measure the production of secondary metabolites, which include polyphenols, flavonoids, and antioxidants. They are all measured in response to weed-induced stress. Total polyphenols, flavonoids, and antioxidants displayed a significant increment in okra when co-cultivated with Lantana or Parthenium. Co-cultivation with both weeds presented synergistic negative effects, like that interaction is characterized by elevated markers of stress, such as reactive oxygen species (ROS) and malondialdehyde (MDA) levels, indicating the compounded adverse effect on the okra plants. Moreover, it points out the effect of the weed's presence on secondary metabolism in okra. Gene expression analysis of two genes, phenylalanine ammonia lyase (PAL) and hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (HQT), which are critical in the phenylpropanoid pathway, validated the role of secondary metabolites in weed-induced stress alleviation. High-performance liquid chromatography (HPLC) analysis shows that CGA has increased significantly in okra plants co-cultivated with weeds. It suggests a plant response to counteract weed-induced stress by enhancing CGA production. Therefore, the study shows the impact of weeds on stress indicators, such as ROS, RNS, and MDA. So, it is observed that stress levels are increased in okra in the weed's presence, as indicated by high lipid and proline content. These findings suggest practical strategies for weed management by enhancing secondary metabolism in crops to improve resilience and productivity under stress conditions caused by invasive weeds.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2751"},"PeriodicalIF":3.2,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143662604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selection of DNA Aptamers Against Parathyroid Hormone for Electrochemical Impedimetric Biosensor System Development.","authors":"Reza Didarian, Saharnaz Bargh, Almina Gülerman, Veli Cengiz Özalp, Özcan Erel, Nimet Yildirim-Tirgil","doi":"10.1002/bab.2745","DOIUrl":"https://doi.org/10.1002/bab.2745","url":null,"abstract":"<p><p>This work presents the pioneering development of an aptamer-based electrochemical biosensor for real-time monitoring of parathyroid hormone (PTH) levels, with a focus on intraoperative assessment during parathyroid surgery. It introduces, for the first time, the selection and characterization of aptamers targeting distinct segments of the PTH peptide. The study demonstrates the feasibility and efficacy of the biosensing platform through a precisely designed experimental framework, including SELEX-based aptamer selection, aptamer-peptide interaction analysis, and biosensor fabrication. The SELEX process yields aptamers with notable binding affinities to different fragments of PTH, with the PTH (53-84) aptamer showing particularly sensitive binding to the hormone's C terminus, allowing for precise PTH analysis. Electrochemical characterization reveals significant changes in electrochemical impedance spectroscopy (EIS) signals upon exposure to varying PTH concentrations, highlighting the sensor's sensitivity and selectivity. The increase in charge transfer resistance (Rct) values with rising PTH concentrations underscores the biosensor's capability to detect PTH-induced structural changes, validating its potential for accurate measurement. The biosensor shows remarkable selectivity in the presence of common interferents in serum samples, ensuring precise PTH detection. Stability assessments over a 45-day storage period demonstrate the biosensor's robustness and long-term reliability, affirming its practical suitability. In summary, the developed aptamer-based biosensor represents a promising tool for sensitive and selective PTH detection, with potential applications in biomedical research and clinical diagnostics, particularly for intraoperative PTH analysis during parathyroidectomy. Continued research and optimization efforts hold promise for enhancing its performance and expanding its utility in diverse healthcare settings.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2745"},"PeriodicalIF":3.2,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143662257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diagnosis Accuracy of Raman Spectroscopy in the Identification of Pathogenic Bacteria.","authors":"Shanshan Mao, Yu Zhang, Chaoqun Chen, Liu Cong, Zuobin Zhu, Zhiyu Xie, Ying Li","doi":"10.1002/bab.2741","DOIUrl":"https://doi.org/10.1002/bab.2741","url":null,"abstract":"<p><p>As an emerging technology, Raman spectroscopy (RS) has been used to identify pathogenic bacteria with excellent performance. The aim of this study was to verify the diagnosis accuracy of RS in identification of pathogenic bacteria. This meta-analysis systematically evaluated the accuracy of RS for identification of pathogenic bacteria. We searched the electronic databases of PubMed and Web of Science to obtain relevant articles; STATA 15.1 was used to analyze all sensitivities, specificies, and their 95% confidence interval (CI). The summary receiver operating characteristic curves (SROC) and area under the curve (AUC) were used to display more performance of RS. Nineteen articles were included according to the inclusion and exclusion criteria. The pooled sensitivity and specificity of RS for the identification of pathogenic bacteria were 0.94 (95% CI, 0.89-0.96) and 0.99 (95% CI, 0.97-0.99). The diagnostic odds ratio (DOR) was 1209 (95% CI, 367-3980), and AUC of SROC was 0.99 (95% CI, 0.98-1.00). For gram-positive bacteria, the sensitivity and specificity of different species ranged from 0.00 to 1.00 and 0.96 to 1.00, with a pooled sensitivity and specificity of 0.96 (95% CI, 0.90-0.98) and 0.99 (95% CI, 0.98-1.00). For gram-negative bacteria, the sensitivity and specificity of different species ranged from 0.30 to 1.00 and 0.92 to 1.00, with a pooled sensitivity and specificity of 0.92 (95% CI, 0.76-0.98) and 0.99 (95% CI, 0.98-1.00). For acid-fast bacteria, the sensitivity and specificity of different species ranged from 0.83 to 1.00 and 0.96 to 1.00, with a pooled sensitivity and specificity of 0.96 (95% CI, 0.84-0.99) and 1.00 (95% CI, 0.96-1.00). RS provides a new method for pathogenic bacteria identification and demonstrates high sensitivity and specificity for most included species.</p>","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":"e2741"},"PeriodicalIF":3.2,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143622987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RETRACTION: Downregulation of SDCBP Inhibits Cell Proliferation and Induces Apoptosis by Regulating PI3K/AKT/mTOR Pathway in Gastric Carcinoma.","authors":"","doi":"10.1002/bab.2733","DOIUrl":"https://doi.org/10.1002/bab.2733","url":null,"abstract":"","PeriodicalId":9274,"journal":{"name":"Biotechnology and applied biochemistry","volume":" ","pages":""},"PeriodicalIF":3.2,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}