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https://www.readbyqxmd.com/read/28738677/quantitation-of-87-proteins-by-nlc-mrm-ms-in-human-plasma-workflow-for-large-scale-analysis-of-biobank-samples
#1
Melinda Rezeli, Karin Sjödin, Henrik Lindberg, Olof Gidlöf, Bertil Lindahl, Tomas Jernberg, Jonas Spaak, David Erlinge, György Marko-Varga
A multiple reaction monitoring (MRM) assay was developed for precise quantitation of 87 plasma proteins including the three isoforms of apolipoprotein E (APOE) associated with cardiovascular diseases using nanoscale liquid chromatography separation and stable isotope dilution strategy. The analytical performance of the assay was evaluated and we found an average technical variation of 4.7 % in 4 - 5 orders of magnitude dynamic range (≈ 0.2 mg/L to 4.5 g/L) from whole plasma digest. Here, we report a complete workflow, including sample processing adapted to 96-well plate format and normalization strategy for large-scale studies...
July 25, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28735491/nanofluidic-allele-specific-digital-pcr-method-for-quantifying-idh1-and-idh2-mutation-burden-in-acute-myeloid-leukemia
#2
Daniel H Wiseman, Tim C P Somervaille
Precise quantitation of allelic burden for a pathogenic mutation has diverse clinical and research applications but can be difficult to achieve with conventional qPCR-based techniques, especially at lower mutant allele frequencies. Digital PCR overcomes many of the limitations of qPCR and can be highly quantitative even for single-nucleotide variants, with distinct advantages over next-generation sequencing approaches. Here we describe a method combining the principles of TaqMan(®)-chemistry SNP genotyping with microfluidic digital PCR to generate a highly sensitive, quantitative allele-specific digital PCR assay for the six most common IDH1 and IDH2 mutations encountered in myeloid malignancy...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28734989/hydroxyvitamin-d-assays-an-historical-perspective-from-deqas
#3
G D Carter, J Berry, R Durazo-Arvizu, E Gunter, G Jones, J Jones, H L J Makin, P Pattni, C T Sempos, P Twomey, E L Williams, S A Wise
Recent years have seen a substantial increase in demand for 25-hydroxyvitamin D (25-OHD) assays. DEQAS (the Vitamin D External Quality Assessment Scheme) has been monitoring the performance of these assays since 1989. The first DEQAS distribution was in June 1989 and results were submitted by 13 laboratories in the UK, two of which used HPLC/UV; the rest used ligand binding assays with a tritium tracer. Inter-laboratory CVs (ALTM) ranged from 29.3% (42.7 nmol/L) to 53.7% (20.0 nmol/L). Currently the scheme has participants in 56 countries using 30 methods or variants of methods...
July 19, 2017: Journal of Steroid Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/28727918/sensitive-and-quantitative-detection-of-anti-poly-ethylene-glycol-peg-antibodies-by-methoxy-peg-coated-surface-plasmon-resonance-sensors
#4
Peng Zhang, Fang Sun, Hsiang-Chieh Hung, Priyesh Jain, Kasey Joanne Leger, Shaoyi Jiang
Pre-existing and induced anti-poly(ethylene glycol) (PEG) antibodies (abs) have been shown to be related with limitation of therapeutic efficacy and reduction in tolerance of several therapeutic agents. However, the current methods to detect anti-PEG abs are tedious and usually lack quantification. A facile, rapid, sensitive, and reliable technique to detect anti-PEG abs is highly desired in both research and clinic settings. In this work, we have presented a surface plasmon resonance (SPR) biosensor technique for the detection of anti-PEG abs and compared three PEG surface chemistries...
July 24, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28724560/accurate-detection-of-methicillin-resistant-staphylococcus-aureus-in-mixtures-utilizing-single-bacterial-duplex-droplet-digital-pcr
#5
Jun Luo, Junhua Li, Hang Yang, Junping Yu, Hongping Wei
Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) is needed to screen MRSA carriers and improve treatment. The current widely used duplex PCR methods are not able to differentiate MRSA from the co-existence of methicillin-susceptible S. aureus (MSSA) and other methicillin-resistant Staphylococci In this study, we aimed to develop a direct method for accurate and rapid detection of MRSA in clinical samples from open environments, such as nasal swab. The new molecular assay is based on detecting the co-occurrence of nuc and mecA markers in a single bacterial cell by utilizing droplet digital PCR (ddPCR) with a chimeric lysin ClyH for cell lysis...
July 19, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28722437/surface-plasmon-resonance-clinical-biosensors-for-medical-diagnostics
#6
Jean-Francois Masson
The design and application of sensors for monitoring biomolecules in clinical samples is a common goal of the sensing research community. Surface plasmon resonance (SPR) and other plasmonic techniques such as localized surface plasmon resonance (LSPR) and imaging SPR are reaching a maturity level sufficient for their application in monitoring biomolecules in clinical samples. In recent years, the first examples for monitoring antibodies, proteins, enzymes, drugs, small molecules, peptides, and nucleic acids in biofluids collected from patients afflicted with a series of medical conditions (Alzheimer's, hepatitis, diabetes, leukemia, and cancers such as prostate and breast cancers, among others) demonstrate the progress of SPR sensing in clinical chemistry...
January 27, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28675028/egfr-and-cd44-dual-targeted-multifunctional-hyaluronic-acid-nanogels-boost-protein-delivery-to-ovarian-and-breast-cancers-in-vitro-and-in-vivo
#7
Jing Chen, Jia Ouyang, Qijun Chen, Chao Deng, Fenghua Meng, Jian Zhang, Ru Cheng, Qing Lan, Zhiyuan Zhong
Protein drugs with intracellular targets like Granzyme B (GrB) have demonstrated great proliferative inhibition activity in cancer cells. Their clinical translation, however, relies on the development of safe, efficient, and selective protein-delivery vehicles. Here, we report that epidermal growth factor receptor (EGFR) and CD44 dual-targeted multifunctional hyaluronic acid nanogels (EGFR/CD44-NGs) boost protein delivery to ovarian and breast cancers in vitro and in vivo. EGFR/CD44-NGs obtained via nanoprecipitation and photoclick chemistry from hyaluronic acid derivatives with tetrazole, GE11 peptide/tetrazole, and cystamine methacrylate groups had nearly quantitative loading of therapeutic proteins like cytochrome C (CC) and GrB, a small size of ca...
July 19, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28665128/discovery-of-the-irreversible-covalent-fgfr-inhibitor-8-3-4-acryloylpiperazin-1-yl-propyl-6-2-6-dichloro-3-5-dimethoxyphenyl-2-methylamino-pyrido-2-3-d-pyrimidin-7-8h-one-prn1371-for-the-treatment-of-solid-tumors
#8
Ken A Brameld, Timothy D Owens, Erik Verner, Eleni Venetsanakos, J Michael Bradshaw, Vernon T Phan, Danny Tam, Kwan Leung, Jin Shu, Jacob LaStant, David G Loughhead, Tony Ton, Dane E Karr, Mary E Gerritsen, David M Goldstein, Jens Oliver Funk
Aberrant signaling of the FGF/FGFR pathway occurs frequently in cancers and is an oncogenic driver in many solid tumors. Clinical validation of FGFR as a therapeutic target has been demonstrated in bladder, liver, lung, breast, and gastric cancers. Our goal was to develop an irreversible covalent inhibitor of FGFR1-4 for use in oncology indications. An irreversible covalent binding mechanism imparts many desirable pharmacological benefits including high potency, selectivity, and prolonged target inhibition...
July 25, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28649510/targeted-screening-of-succinic-semialdehyde-dehydrogenase-deficiency-ssadhd-employing-an-enzymatic-assay-for-%C3%AE-hydroxybutyric-acid-ghb-in-biofluids
#9
C Wernli, S Finochiaro, C Volken, H Andresen-Streichert, A Buettler, D Gygax, G S Salomons, E E Jansen, G R Ainslie, K R Vogel, K M Gibson
HYPOTHESIS: An enzymatic assay for quantification of γ-hydroxybutyric acid (GHB) in biofluids can be employed for targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) in selected populations. RATIONALE: We used a two-tiered study approach, in which the first study (proof of concept) examined 7 urine samples derived from patients with SSADHD and 5 controls, and the second study (feasibility study) examined a broader sample population of patients and controls, including plasma...
June 2017: Molecular Genetics and Metabolism Reports
https://www.readbyqxmd.com/read/28647526/national-survey-of-adult-and-pediatric-reference-intervals-in-clinical-laboratories-across-canada-a-report-of-the-cscc-working-group-on-reference-interval-harmonization
#10
Khosrow Adeli, Victoria Higgins, David Seccombe, Christine P Collier, Cynthia Balion, George Cembrowski, Allison A Venner, Julie Shaw
OBJECTIVE: Reference intervals are widely used decision-making tools in laboratory medicine, serving as health-associated standards to interpret laboratory test results. Numerous studies have shown wide variation in reference intervals, even between laboratories using assays from the same manufacturer. Lack of consistency in either sample measurement or reference intervals across laboratories challenges the expectation of standardized patient care regardless of testing location. Here, we present data from a national survey conducted by the Canadian Society of Clinical Chemistry (CSCC) Reference Interval Harmonization (hRI) Working Group that examines the variation in laboratory reference sample measurements, as well as pediatric and adult reference intervals currently used in clinical practice across Canada...
June 21, 2017: Clinical Biochemistry
https://www.readbyqxmd.com/read/28643351/quality-specifications-of-routine-clinical-chemistry-methods-based-on-sigma-metrics-in-performance-evaluation
#11
Jun Xia, Su-Feng Chen, Fei Xu, Yong-Lie Zhou
INTRODUCTION: Sigma metrics were applied to evaluate the performance of 20 routine chemistry assays, and individual quality control criteria were established based on the sigma values of different assays. METHODS: Precisions were expressed as the average coefficient variations (CVs) of long-term two-level chemistry controls. The biases of the 20 assays were obtained from the results of trueness programs organized by National Center for Clinical Laboratories (NCCL, China) in 2016...
June 23, 2017: Journal of Clinical Laboratory Analysis
https://www.readbyqxmd.com/read/28624482/nmr-quantification-of-trimethylamine-n-oxide-in-human-serum-and-plasma-in-the-clinical-laboratory-setting
#12
Erwin Garcia, Justyna Wolak-Dinsmore, Zeneng Wang, Xinmin S Li, Dennis W Bennett, Margery A Connelly, James D Otvos, Stanley L Hazen, Elias J Jeyarajah
BACKGROUND AND OBJECTIVES: Trimethylamine-N-oxide (TMAO) produced by gut microbiota metabolism of dietary choline and carnitine has been shown to be associated with increased risk of cardiovascular disease (CVD) and to provide incremental clinical prognostic utility beyond traditional risk factors for assessing a patient's CVD risk. The aim of this study was to develop an automated nuclear magnetic resonance (NMR) spectroscopy assay for quantification of TMAO concentration in serum and plasma using a high-throughput NMR clinical analyzer...
June 15, 2017: Clinical Biochemistry
https://www.readbyqxmd.com/read/28623087/highly-multiplex-real-time-pcr-based-screening-for-blood-borne-pathogens-on-an-openarray-platform
#13
Elena Grigorenko, Carolyn Fisher, Sunali Patel, Valerie Winkelman, Phillip Williamson, Caren Chancey, Germán Añez, Maria Rios, Victoria Majam, Sanjai Kumar, Robert Duncan
Molecular diagnostics are increasingly used in the blood bank industry. A device that can combine simultaneous detection of multiple targets with the flexibility of inclusion of emerging pathogens is desirable for testing blood products. A highly multiplexed blood-borne pathogen panel (BBPP) using dual-label probe chemistry (TaqMan assays) was developed for simultaneous detection and discrimination of 17 viral pathogens in human plasma samples and 13 bacterial and protozoan pathogens in human blood samples on the OpenArray platform...
June 12, 2017: Journal of Molecular Diagnostics: JMD
https://www.readbyqxmd.com/read/28603416/successful-in-vivo-hyperthermal-therapy-toward-breast-cancer-by-chinese-medicine-shikonin-loaded-thermosensitive-micelle
#14
Yonghua Su, Nian Huang, Di Chen, Li Zhang, Xia Dong, Yun Sun, Xiandi Zhu, Fulei Zhang, Jie Gao, Ying Wang, Kexing Fan, Puichi Lo, Wei Li, Changquan Ling
The Chinese traditional medicine Shikonin is an ideal drug due to its multiple targets to tumor cells. But in clinics, improving its aqueous solubility and tumor accumulation is still a challenge. Herein, a copolymer with tunable poly(N-isopropylacrymaide) and polylactic acid block lengths is designed, synthesized, and characterized in nuclear magnetic resonance. The corresponding thermosensitive nanomicelle (TN) with well-defined core-shell structure is then assembled in an aqueous solution. For promoting the therapeutic index, the physical-chemistry properties of TNs including narrow size, low critical micellar concentration, high serum stability, tunable volume phase transition temperature (VPTT), high drug-loading capacity, and temperature-controlled drug release are systematically investigated and regulated through the fine self-assembly...
2017: International Journal of Nanomedicine
https://www.readbyqxmd.com/read/28598594/tethering-growth-factors-to-collagen-surfaces-using-copper-free-click-chemistry-surface-characterization-and-in-vitro-biological-response
#15
Hyun Jong Lee, Gabriella M Fernandes-Cunha, Ilham Putra, Won-Gun Koh, David Myung
Surface modifications with tethered growth factors have mainly been applied to synthetic polymeric biomaterials in well-controlled, acellular settings, followed by seeding with cells. The known bio-orthogonality of copper-free click chemistry provides an opportunity to not only use it in vitro to create scaffolds or pro-migratory tracks in the presence of living cells, but also potentially apply it to living tissues directly as a coupling modality in situ. In this study, we studied the chemical coupling of growth factors to collagen using biocompatible copper-free click chemistry and its effect on the enhancement of growth factor activity in vitro...
July 19, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28594875/analytical-evaluation-of-three-enzymatic-assays-for-measuring-total-bile-acids-in-plasma-using-a-fully-automated-clinical-chemistry-platform
#16
Elisa Danese, Gian Luca Salvagno, Davide Negrini, Giorgio Brocco, Martina Montagnana, Giuseppe Lippi
BACKGROUND: Although the clinical significance of measuring bile acids concentration in plasma or serum has been recognized for long in patients with hepatobiliary disease and/or bile acid malabsorption, the reference separation techniques are expensive and mostly unsuitable for early diagnosis and for measuring large volumes of samples. Therefore, this study was aimed to evaluate the analytical performance of three commercial enzymatic techniques for measuring total bile acids in plasma using a fully-automated clinical chemistry platform...
2017: PloS One
https://www.readbyqxmd.com/read/28584208/establishment-of-a-novel-method-for-screening-epidermal-growth-factor-receptor-tyrosine-kinase-inhibitor-resistance-mutations-in-lung-cancer
#17
Hong-Xia Tian, Xu-Chao Zhang, Zhen Wang, Jin-Ji Yang, Wei-Bang Guo, Zhi-Hong Chen, Yi-Long Wu
BACKGROUND: Drug resistance to targeted therapies occurs in lung cancer, and resistance mechanisms related to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are continuously being discovered. We aimed to establish a novel method for highly parallel multiplexed detection of genetic mutations related to EGFR TKI-resistant lung cancer using Agena iPLEX chemistry and matrix-assisted laser desorption ionization time-of-flight analysis on the MassARRAY mass spectrometry platform...
June 20, 2017: Chinese Medical Journal
https://www.readbyqxmd.com/read/28583666/safety-evaluation-of-water-soluble-palm-fruit-bioactives
#18
Barry S Lynch, Spencer West, Ashley Roberts
Water-soluble palm fruit bioactives, derived from the aqueous stream of palm oil processing, have shown anti-diabetogenic effects in rodent models. To assess the safety of potential incorporation of this polyphenol-containing material in food, in vitro bacterial reverse mutation and in vitro chromosome aberration assays were conducted along with a 90-day subchronic toxicity study in Sprague-Dawley rats. Water-soluble palm fruit bioactives were inactive in the Ames and in vitro chromosome aberration assays up to the limit doses of 5000 μg/plate and 5000 μg/mL, respectively...
June 2, 2017: Regulatory Toxicology and Pharmacology: RTP
https://www.readbyqxmd.com/read/28558755/evaluation-of-a-species-specific-c-reactive-protein-assay-for-the-dog-on-the-abx-pentra-400-clinical-chemistry-analyzer
#19
Sarah Hindenberg, Stefanie Klenner-Gastreich, Nicole Kneier, Sabine Zielinsky, Kris Gommeren, Natali Bauer, Andreas Moritz
BACKGROUND: A canine-specific immunoturbidimetric CRP assay, Gentian Canine CRP Immunoassay) with species-specific controls and calibrators was introduced and recently evaluated on the clinical chemistry analyzer Abbott Architect c4000 as well as on the Olympus AU600. Aims of our study were 1) to independently evaluate the canine-specific CRP assay on the ABX Pentra 400 clinical chemistry analyzer in comparison to the previously validated human-based immunoturbidimetric assay (Randox Canine CRP assay) and 2) to assess the impact of different sample types (serum versus heparinized plasma) on the results...
May 30, 2017: BMC Veterinary Research
https://www.readbyqxmd.com/read/28535882/versatile-fabrication-of-paper-based-microfluidic-devices-with-high-chemical-resistance-using-scholar-glue-and-magnetic-masks
#20
Thiago M G Cardoso, Fabrício R de Souza, Paulo T Garcia, Denilson Rabelo, Charles S Henry, Wendell K T Coltro
Simple methods have been developed for fabricating microfluidic paper-based analytical devices (μPADs) but few of these devices can be used with organic solvents and/or aqueous solutions containing surfactants. This study describes a simple fabrication strategy for μPADs that uses readily available scholar glue to create the hydrophobic flow barriers that are resistant to surfactants and organic solvents. Microfluidic structures were defined by magnetic masks designed with either neodymium magnets or magnetic sheets to define the patter, and structures were created by spraying an aqueous solution of glue on the paper surface...
June 29, 2017: Analytica Chimica Acta
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