Zi-qing Wang , Li-ping Hao , Zi-xuan Meng , Hao-ran Zhang , Wei-jun Kang , Lian-feng Ai
{"title":"Simultaneous determination of 25(OH)D2, 25(OH)D3 and 1α,25(OH)2D3 in human serum by derivatization-liquid chromatography-tandem mass spectrometry","authors":"Zi-qing Wang , Li-ping Hao , Zi-xuan Meng , Hao-ran Zhang , Wei-jun Kang , Lian-feng Ai","doi":"10.1016/j.ab.2025.115821","DOIUrl":"10.1016/j.ab.2025.115821","url":null,"abstract":"<div><div>An LC-MS/MS method was developed and validated for simultaneously quantifying 25-hydroxyvitamin D2, D3, and 1α,25-dihydroxyvitamin D3 in human serum. Protein in 200 μL serum was precipitated with acetonitrile. After centrifugation, the metabolites were derivatized using (diacetoxyiodo)benzene (PIA) and 4-(4-dimethylaminophenyl)-1,2,4-triazolidine-3,5-dione (DMAT) and then quantified by the LC-MS/MS system. The limits of detection (LODs) for the three substances were 10, 10, and 5 pg/mL, and the limits of quantification (LOQs) were 20, 20, and 10 pg/mL. The standard curves for these compounds showed linear regression coefficients (R2)>0.998 over specific concentration ranges. Recoveries were 94.36 %–102.34 % for 25(OH)D2, 92.43 %–103.41 % for 25(OH)D3, and 88.98 %–94.36 % for 1α,25(OH)2D3. The mean serum levels in 109 subjects (consisting of 61 healthy adult males and 59 healthy adult females) were 2.0 ± 1.5 ng/ml (25(OH)D2), 16.4 ± 6.1 ng/ml (25(OH)D3) and 36.6 ± 15.1 pg/ml (1α, 25(OH)2D3).Derivatization of vitamin D metabolites using PIA and DMAT is useful for rapidly determining the serum 25(OH) D2, 25(OH) D3 and 1α,25(OH)2D3 concentrations simultaneously in human serum.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115821"},"PeriodicalIF":2.6,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143514352","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}
Gabriele Frost , Christian Lutz , Andreas Schwiertz , Hermann Huber , Albert Missbichler
{"title":"Total histamine degrading capacity: Correlation to stool histamine and its usefulness in supporting the diagnosis of histamine intolerance","authors":"Gabriele Frost , Christian Lutz , Andreas Schwiertz , Hermann Huber , Albert Missbichler","doi":"10.1016/j.ab.2025.115819","DOIUrl":"10.1016/j.ab.2025.115819","url":null,"abstract":"<div><div>The relatively new test (European Patent granted 2022) for determining the total histamine degradation capacity [Totale Histamin Abbau Kapazität/Total Histamine Degrading Capacity] THAK®/THDC of a serum sample has been compared in various settings with the individual clinical history and the levels of histamine in stool. The measurement of THAK can be carried out regardless of the current status of the patient (independent of any acute symptoms), therefore THAK test is a useful and very reliable instrument for supporting the diagnosing histamine intolerance. As soon as the THAK falls below a threshold value the patient is most likely to suffer from HIT, even if the intensity of the symptoms is low. Low THAK values correspond with a significant increase of histamine in stool. Results were evaluated according to both age and gender, no difference between the sexes were found. However, there is a noticeable dependency on age, although this is not diagnostically relevant.</div><div>THAK® is a registered trademark of Frost Diagnostika.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"702 ","pages":"Article 115819"},"PeriodicalIF":2.6,"publicationDate":"2025-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143490444","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}
Peng Zhang , Yi Liu , Xinyun Zhou , Yinyu Chen , Liudi Pan , Feiyi Xu , Xiaolong Zhang , Lisong Shen , Hui Chen
{"title":"Evaluation of a multiplexed assay for six urinary proteins based on liquid microarray analysis technology","authors":"Peng Zhang , Yi Liu , Xinyun Zhou , Yinyu Chen , Liudi Pan , Feiyi Xu , Xiaolong Zhang , Lisong Shen , Hui Chen","doi":"10.1016/j.ab.2025.115820","DOIUrl":"10.1016/j.ab.2025.115820","url":null,"abstract":"<div><div>Proteinuria serves as a critical indicator in the progression of chronic kidney disease. The quantification of urinary protein is instrumental in diagnosing kidney disease and monitoring therapeutic efficacy. Nephelometry and turbidimetric immunoassay are predominantly employed for the quantitative detection of various urinary proteins. We developed a quantitative detection method for six proteins in urine utilizing the flow fluorescence luminescence method on the Luminex®200™ detection platform. This study aims to evaluate the clinical performance and accuracy of the method. The sensitivity, precision, recovery, linearity, and interference metrics for the method meet the needs of clinical applications. The correlation coefficient (r) between the results of the method and nephelometry immunoassay on SIEMENS BNII platform ranged from 0.9375 to 0.9847, indicating a strong linear relationship. The results of the Bland-Altman analysis indicated that the systematic bias between the two methods was minimal. Additionally, Passing-Bablok regression analysis, both including and excluding outliers, was conducted on the clinical trial data. The findings demonstrated consistency between the two methods, with the expected confidence interval of bias remaining within the allowable error limits, indicating no significant discrepancy. This urine six protein quantification method demonstrates satisfactory performance, meeting the standards necessary for medical diagnosis and clinical application.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115820"},"PeriodicalIF":2.6,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143476206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Huda Abdulsahib Sabbar, Sarah Kadhim Al-Hayali, Abdulhadi Al-Janabi
{"title":"A drop-like optical fiber salivary pH sensor: A rapid and real-time detection of periodontal disease","authors":"Huda Abdulsahib Sabbar, Sarah Kadhim Al-Hayali, Abdulhadi Al-Janabi","doi":"10.1016/j.ab.2025.115818","DOIUrl":"10.1016/j.ab.2025.115818","url":null,"abstract":"<div><div>A real-time and rapid detection of <strong>s</strong>alivary pH for periodontal diagnosis is an emerging point-of-care (POC) technology trend. The precise and non-invasive sensitive POC devices, which could be used as chair-side tools to improve clinical dental inspections, are essential for an excellent periodontal diagnosis. In this paper, we report on developing a POC technology based on optical fiber sensors as a promising marker, to the best of our knowledge, to detect the role of salivary pH in periodontitis diagnosis and support dentists’ inspection. The optical fiber sensor based on a drop-like structure was fabricated by bending single-mode fiber (SMF) into a section of a thin capillary tube. In this work, firstly, the performance of the proposed sensor towards different pH levels ranging from 1 to 7 was experimentally investigated. Experimentations showed that the pH sensor responded to various pH levels with a calibration curve sensitivity of −2.075 nm/pH and a high linearity of 0.985. Then, 66 patients were enrolled in this study and divided into two groups according to clinical outcomes: Group A with clinically healthy periodontium and Group B with chronic periodontitis. The real saliva from each patient was collected, and the salivary pH was evaluated immediately using the aforementioned proposed sensor. The experimental results show that Group A and Group B salivary pH mapping ranged from 6.5 to 7.3 and 5.5 to 6.4, respectively. The proposed sensor offers substantial practical benefits, providing a rapid response time of less than 2 s, instantaneous, noninvasive, and easily monitored tool for the early detection and diagnosis of periodontal health status.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115818"},"PeriodicalIF":2.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143465095","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}
Ali Alqahtani , Taha Alqahtani , Adel Al Fatease , Ahmed A. Almrasy
{"title":"Development and validation of a graphene quantum dot-based sensor for abacavir quantification via fluorescence quenching","authors":"Ali Alqahtani , Taha Alqahtani , Adel Al Fatease , Ahmed A. Almrasy","doi":"10.1016/j.ab.2025.115817","DOIUrl":"10.1016/j.ab.2025.115817","url":null,"abstract":"<div><div>Human immunodeficiency virus (HIV) is a global health concern, and the development of effective methods for the detection and quantification of antiretroviral drugs is crucial for monitoring therapeutic efficacy and patient compliance. Herein, we present a novel approach utilizing graphene quantum dots (GQDs) for the determination of abacavir, a widely used nucleoside reverse transcriptase inhibitor in the treatment of HIV. The sensing mechanism was investigated through Stern-Volmer analysis, thermodynamics studies, and density functional theory calculations, which revealed the strong binding interactions between GQDs and abacavir <em>via</em> a static quenching process. Factors affecting the analytical performance, such as pH, GQDs concentration, and incubation time, were systematically optimized to achieve a linear detection range of 100–1000 ng/mL with a low detection limit of 17.49 ng/mL. The method was validated in accordance with ICH guidelines demonstrating excellent linearity, accuracy, precision, robustness and selectivity making it suitable for the quantification of abacavir in pharmaceutical formulations and biological samples. The method also demonstrated higher sensitivity and more environmentally friendly characteristics when compared to previously reported chromatographic techniques, showcasing the potential of GQDs as a superior alternative for the traditional detection approaches of pharmaceutical compounds.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115817"},"PeriodicalIF":2.6,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143429419","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}
Jiani Li , Min Ju , Danni Zheng , Huihui Wan , Yuming Sun , Jinfeng Zhao , Lina Zhou , Qingxin Yin , Wei Kang , Yuming Song , Song Xue
{"title":"Simultaneous analysis of 7 key mevalonate pathway intermediates using liquid chromatography-orbitrap mass spectrometry","authors":"Jiani Li , Min Ju , Danni Zheng , Huihui Wan , Yuming Sun , Jinfeng Zhao , Lina Zhou , Qingxin Yin , Wei Kang , Yuming Song , Song Xue","doi":"10.1016/j.ab.2025.115816","DOIUrl":"10.1016/j.ab.2025.115816","url":null,"abstract":"<div><div>The mevalonate (MVA) pathway is a central metabolic route that converts mevalonate into isoprenoids, which are of crucial significance in various cellular processes. As no systematic approach had been developed and validated for the simultaneous estimation of these intermediate metabolites within the MVA pathway. In this research, a method was developed for the concurrent determination of 7 key intermediate metabolites in the MVA pathway based on ultra-high performance liquid chromatography (UPLC) coupled with a quadrupole/electrostatic field orbitrap high-resolution mass spectrometry (HRMS). The developed UPLC-HRMS method based on the hydrophilic interaction liquid chromatography (HILIC) separation mode was successfully applied to the quantitative analysis of relevant key intermediate metabolites in wild type <em>Escherichia coli</em> (<em>E. coli</em>) BL21 (DE3) and the recombinant <em>E. coli</em> BL21 (DE3) system constructed by introducing all the genes of the MVA pathway into the strain. The above results can offer a reference for further studies on the MVA pathway.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115816"},"PeriodicalIF":2.6,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143432215","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}
Yiyi Hu , Yunchuan Li , Haodan Zhu , Dandan Wang , Junming Zhou , Bin Li
{"title":"Screening and comparative study of four anti-PEDV candidate drugs in vitro","authors":"Yiyi Hu , Yunchuan Li , Haodan Zhu , Dandan Wang , Junming Zhou , Bin Li","doi":"10.1016/j.ab.2025.115804","DOIUrl":"10.1016/j.ab.2025.115804","url":null,"abstract":"<div><div>Piglets afflicted with Porcine Epidemic Diarrhea Virus (PEDV) experience severe diarrhea and elevated death rates, leading to substantial financial losses in the pig farming sector. The objective of this study is to investigate the impact of polyphenols on PEDV within Vero cells by utilizing different methodologies to evaluate their anti-PEDV effect. By producing 30 polyphenols, we have discovered that No.2, No.5, No.7 and No.17 exhibit properties that make them effective against PEDV, serving as potential drugs. The findings showed that in a clear dose-dependent manner, the mRNA levels of PEDV were significantly inhibited in the high, middle, and low-dose groups of No.2, No.5, No.7 and No.17, when compared to the PEDV control. The four tested polyphenols significantly inhibited the levels of PEDV N contents and viral titers. Furthermore, concentration of cytotoxicity 50 % (CC<sub>50</sub>) values for No.2, No.5, No.7 and No.17 polyphenols were 42.84 μM, 16.78 μM, 39.01 μM and 28.86 μM, respectively, demonstrating the safety of these medications in clinical environments. While they exert their anti-PEDV activity all specifically at the viral replication stage. Collectively, these findings indicate that the four examined polyphenols could efficiently modulate the immune response against PEDV and hold promise for utilization in antiviral treatments.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115804"},"PeriodicalIF":2.6,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143420257","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":"Simulation of absorption spectra of native and unfolded proteins","authors":"Tyson J. Pilkington, Stefan Siemann","doi":"10.1016/j.ab.2025.115803","DOIUrl":"10.1016/j.ab.2025.115803","url":null,"abstract":"<div><div>Protein concentrations are routinely determined using the absorbance measured at 280 nm and Beer-Lambert's law. However, traditional single-wavelength approaches may be inferior to a multi-wavelength analysis of complete spectra given the larger amount of data that can be processed. Hence, the current study was aimed at simulating protein UV spectra (from 250 to 350 nm) with a view to more accurately estimate protein concentrations. We demonstrate that the spectra of unfolded proteins are well-simulated using primary sequence data and the wavelength-dependent molar absorption coefficients of <span>l</span>-cystine, <span>l</span>-phenylalanine, <em>N</em>-acetyl-<span>l</span>-tyrosinamide and <em>N</em>-acetyl-<span>l</span>-tryptophanamide (the latter two serving as L-Tyr and L-Trp model compounds). Alternatively, simulations can be performed with the coefficients of the Tyr and Trp mimics replaced by a pseudo-Voigt (pV) function, which mathematically fully describes the spectra of these model compounds. Furthermore, a pV function, generated from the analysis of the spectra of 14 proteins, can be utilized to simulate the spectral contributions of Tyr and Trp in native proteins with a reasonable degree of accuracy. A Microsoft Excel-based multi-wavelength fitting routine can be employed to simulate the spectra of proteins and compare them with those experimentally recorded, thereby facilitating the determination of protein concentrations.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115803"},"PeriodicalIF":2.6,"publicationDate":"2025-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143424442","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}
Sarah Alrubia , Amsha S. Alsegiani , Nourah Z. Alzoman, Ibrahim A. Darwish
{"title":"A sensitive HPLC method with fluorescence detection for quantification of pemigatinib in human plasma samples and its in-vivo application to pharmacokinetic study in rats","authors":"Sarah Alrubia , Amsha S. Alsegiani , Nourah Z. Alzoman, Ibrahim A. Darwish","doi":"10.1016/j.ab.2025.115805","DOIUrl":"10.1016/j.ab.2025.115805","url":null,"abstract":"<div><div>Cholangiocarcinoma is a lethal tumour of the bile ducts. Cholangiocarcinoma fibroblast growth factor receptor gene fusions forms can be targeted by Pemigatinib (PGT). PGT a recently approved kinase inhibitor by the US-FDA, has its approval accelerated due to the disease viciousness. Development of a sensitive yet available and economical analytical platform to quantify PGT in human plasma is genuinely needed. Enabling monitoring of the therapeutic plan, hence, ensuring the drug efficacy and safety through pharmacokinetic studies. High-performance liquid chromatography with fluorescence detector (HPLC-FD) method is proposed using the native fluorescence of PGT. PGT and seliciclib (internal standard) chromatographic conditions optimisation, revealed favourable use of isocratic mobile phase consisting of methanol:ammonium acetate buffer (70:30v/v, pH5.0) pumped into C18-column (150 mm length × 4.6 mm internal diameter, 5 μm particle size), at 1 mL min<sup>−1</sup> flow rate. PGT and seliciclib fluorescence excitation and emission were measured at 280 and 360 nm, respectively. Validation of the HPLC-FD method was processed based on the International Council for Harmonization guidelines. The method linearity range was 5–300 ng mL<sup>−1</sup>. The limits of detection and quantification were 2.8 and 8.5 ng mL<sup>−1</sup>, respectively. High precision and accuracy indicated by relative standard deviation ≤ 5.2, and recovery values of 95.4–102.2 %. Evident of the method greenness was verified by green analytical chemistry metric tools. HPLC-FD method was successfully applied to study PGT pharmacokinetics in rats. In conclusion, this study introduces a reliable analytical method of PGT in plasma for routine use in therapeutic drug monitoring for quality assurance and clinical follow-up.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115805"},"PeriodicalIF":2.6,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143412918","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}
Vadim S. Pokrovsky , Louay Abo Qoura , Aleksei A. Tikhonov , Alla Yu Rubina , Nikolai E. Kushlinskii
{"title":"Multiplex analysis of ovarian cancer patients using glycan microarray","authors":"Vadim S. Pokrovsky , Louay Abo Qoura , Aleksei A. Tikhonov , Alla Yu Rubina , Nikolai E. Kushlinskii","doi":"10.1016/j.ab.2025.115806","DOIUrl":"10.1016/j.ab.2025.115806","url":null,"abstract":"<div><div>Investigation of tumor-associated glycan antigens (TAGs) could be helpful for the development of sensitive cancer diagnostics and novel therapies. Glycan microarrays are effective methods for analyzing glycans and anti-glycan antibodies, which are immobilized arrays of oligo- or poly-saccharides on different substrates, making them a promising class of oncological biomarkers. Blood serum samples from patients (n = 203) with ovarian cancer (OvaCan) and healthy volunteers were analyzed using a glycan microarray containing 63 immobilized glycans to determine changes in anti-glycan IgG and IgM antibody profiles in OvaCan. Levels of anti-glycan IgG and IgM antibodies in OvaCan statistically differed from levels in healthy donors: the most prominent statistically significant difference for anti-glycan IgG antibodies was found for 6-<em>O</em>-su-Le<sup>c</sup> (AUC = 0.657, Se = 48.0 %, and Sp = 73.3 %). The AUC values for certain glycans investigated in diagnosing OvaCan indicated a fingerprint consisting of IgM antibodies to specific glycans, and the most specific anti-glycan IgM antibodies were Le<sup>y</sup> (AUC = 0.625, Se = 98.0 % and Sp = 45.0 %). The potential of these serological biomarkers to distinguish between OvaCan and other malignancies is still an unresolved issue that requires more large-scale studies to confirm and validate the use of these biomarkers in the diagnosis of different types of cancer.</div></div>","PeriodicalId":7830,"journal":{"name":"Analytical biochemistry","volume":"701 ","pages":"Article 115806"},"PeriodicalIF":2.6,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143412919","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}