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https://www.readbyqxmd.com/read/29339216/in-vivo-biotinylated-calpastatin-improves-the-affinity-purification-of-human-m-calpain
#1
Hung Huy Nguyen, Alexander N Volkov, Guy Vandenbussche, Peter Tompa, Kris Pauwels
Recently we established a novel affinity purification method for calpain by exploiting the specific and reversible binding properties of its intrinsically disordered protein inhibitor, calpastatin. The immobilization strategy relied on the strength and specificity of the biotin - streptavidin interaction. Here, we report an improved and optimized method that even enables the general applicability of in vivo biotinylated (intrinsically disordered) proteins in any affinity capture strategy. Since in vitro chemical biotinylation is only accomplished with reagents that lack exact site specificity, it can not only cause sample heterogeneity but it can also hamper the functionality of the biotinylated molecules...
January 12, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29338842/real-time-nanopore-based-recognition-of-protein-translocation-success
#2
David P Hoogerheide, Philip A Gurnev, Tatiana K Rostovtseva, Sergey M Bezrukov
A growing number of new technologies are supported by a single- or multi-nanopore architecture for capture, sensing, and delivery of polymeric biomolecules. Nanopore-based single-molecule DNA sequencing is the premier example. This method relies on the uniform linear charge density of DNA, so that each DNA strand is overwhelmingly likely to pass through the nanopore and across the separating membrane. For disordered peptides, folded proteins, or block copolymers with heterogeneous charge densities, by contrast, translocation is not assured, and additional strategies to monitor the progress of the polymer molecule through a nanopore are required...
January 12, 2018: Biophysical Journal
https://www.readbyqxmd.com/read/29325035/amplicon-based-next-generation-sequencing-of-plasma-cell-free-dna-for-detection-of-driver-and-resistance-mutations-in-advanced-non-small-cell-lung-cancer
#3
N Guibert, Y Hu, N Feeney, Y Kuang, V Plagnol, G Jones, K Howarth, J F Beeler, C P Paweletz, G R Oxnard
Background: Genomic analysis of plasma cell-free DNA is transforming lung cancer care, however available assays are limited by cost, turnaround time, and imperfect accuracy. Here we study amplicon-based plasma next-generation sequencing (NGS), rather than hybrid-capture-based plasma NGS, hypothesizing this would allow sensitive detection and monitoring of driver and resistance mutations in advanced non-small cell lung cancer (NSCLC). Methods: Plasma samples from patients with NSCLC and a known targetable genotype (EGFR, ALK/ROS1 and other rare genotypes) were collected while on therapy and analyzed, blinded to tumor genotype...
January 9, 2018: Annals of Oncology: Official Journal of the European Society for Medical Oncology
https://www.readbyqxmd.com/read/29321673/genomic-analysis-of-family-data-reveals-additional-genetic-effects-on-intelligence-and-personality
#4
W David Hill, Ruben C Arslan, Charley Xia, Michelle Luciano, Carmen Amador, Pau Navarro, Caroline Hayward, Reka Nagy, David J Porteous, Andrew M McIntosh, Ian J Deary, Chris S Haley, Lars Penke
Pedigree-based analyses of intelligence have reported that genetic differences account for 50-80% of the phenotypic variation. For personality traits these effects are smaller, with 34-48% of the variance being explained by genetic differences. However, molecular genetic studies using unrelated individuals typically report a heritability estimate of around 30% for intelligence and between 0 and 15% for personality variables. Pedigree-based estimates and molecular genetic estimates may differ because current genotyping platforms are poor at tagging causal variants, variants with low minor allele frequency, copy number variants, and structural variants...
January 10, 2018: Molecular Psychiatry
https://www.readbyqxmd.com/read/29313657/spatial-resolved-photoelectrochemical-biosensing-array-based-on-cds-g-c3n4-heterojunction-a-universal-immunosensing-platform-for-accurate-detection
#5
Yu-Xiang Dong, Juntao Cao, Bing Wang, Shu-Hui Ma, Yan Ming Liu
The detection of biomarkers with high sensitivity and accuracy in real biosamples remains challenging. Herein, a universal spatial-resolved photoelectrochemical (PEC) ratiometry for biodetection of prostate specific antigen (PSA) as model biomarker was designed for the first time based on dual-electrodes array modified by CdS@g-C3N4 heterojunction coupled with CuS quantum dots (QDs) as signal amplification tags. Specifically, a new kind of photoactive material, CdS@g-C3N4 p-n heterojunction with highly photoelectric conversion efficiency and good chemical stability, was synthesized and immobilized on two spatial-resolved electrodes (WE1 and WE2)...
January 9, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29311362/novel-transcriptional-activity-and-extensive-allelic-imbalance-in-the-human-mhc-region
#6
Elizabeth Gensterblum-Miller, Weisheng Wu, Amr H Sawalha
The MHC region encodes HLA genes and is the most complex region in the human genome. The extensively polymorphic nature of the HLA hinders accurate localization and functional assessment of disease risk loci within this region. Using targeted capture sequencing and constructing individualized genomes for transcriptome alignment, we identified 908 novel transcripts within the human MHC region. These include 593 novel isoforms of known genes, 137 antisense strand RNAs, 119 novel long intergenic noncoding RNAs, and 5 transcripts of 3 novel putative protein-coding human endogenous retrovirus genes...
January 8, 2018: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/29310232/electrochemical-aptasensor-for-multi-antibiotics-detection-based-on-endonuclease-and-exonuclease-assisted-dual-recycling-amplification-strategy
#7
Shengfeng Huang, Ning Gan, Tianhua Li, You Zhou, Yuting Cao, Youren Dong
An ultrasensitive electrochemical aptasensor for multiplex antibiotics detection based on endonuclease and exonuclease assisted dual recycling amplification strategy was proposed. Kanamycin and chloramphenicol were selected as candidates. Firstly, aptamers of the antibiotics were immobilized on bar A and then binding with their endonuclease labeled complementary DNA strands to construct enzyme-cleavage probes. Secondly, The nano zirconium-metal organic framework (NMOF) particles with 1,4-benzene-dicarboxylate (BDC) as linker was defined as UiO-66...
March 1, 2018: Talanta
https://www.readbyqxmd.com/read/29303555/an-optical-biosensor-based-quantification-of-microcystin-synthetase-a-gene-early-warning-of-toxic-cyanobacterial-blooming
#8
Jinchuan Liu, Xiaohong Zhou, Hanchang Shi
Monitoring and controlling of toxic cyanobacterial strains, which can produce microcystins, is critical to protect human and ecological health. We herein reported an optical biosensor-based quantification of microcystin synthetase A (mcyA) gene so as to discriminate microcystins-producing from non-producing strains. In this assay, the mcyA-specific ssDNA probes were designed in silico with an online tool and then synthesized to be covalently immobilized on an optical fiber surface. Production of fluorescently modified target DNA fragment amplicons was accomplished through the use of Cy5-tagged deoxycytidine triphosphates (dCTPs) in the polymerase chain reaction (PCR) method, which resulted in copies with internally labelled multiple sites per DNA molecule and delivered the great sensitivity...
January 5, 2018: Analytical Chemistry
https://www.readbyqxmd.com/read/29303554/a-serological-point-of-care-test-for-the-detection-of-igg-antibodies-against-ebola-virus-in-human-survivors
#9
Polina Brangel, Ariel Sobarzo, Claudio Parolo, Benjamin S Miller, Philip D Howes, Sigal Gelkop, Julius J Lutwama, John M Dye, Rachel A McKendry, Leslie Lobel, Molly M Stevens
Ebola virus disease causes widespread and highly fatal epidemics in human populations. Today, there is still great need for point-of-care tests for diagnosis, patient management and surveillance, both during and post outbreaks. We present a point-of-care test comprising an immunochromatographic strip and a smartphone reader, which detects and semiquantifies Ebola-specific antibodies in human survivors. We developed a Sudan virus glycoprotein monoplex platform and validated it using sera from 90 human survivors and 31 local noninfected controls...
January 5, 2018: ACS Nano
https://www.readbyqxmd.com/read/29289826/a-sandwich-type-electrochemical-immunoassay-for-ultrasensitive-detection-of-non-small-cell-lung-cancer-biomarker-cyfra21-1
#10
Yan Zeng, Jing Bao, Yanan Zhao, Danqun Huo, Mei Chen, Yanli Qi, Mei Yang, Huanbao Fa, Changjun Hou
Many studies confirm that the aberrant expression of Cytokeratin 19 fragment 21-1 (CYFRA21-1) is highly correlated with non-small cell lung cancer (NSCLC), especially for squamous cell carcinoma. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to test CYFRA21-1 selectively and sensitively. The proposed immunosensor fabricated by three-dimensional graphene (3D-G), chitosan (CS) and glutaraldehyde (GA) composite on the glass carbon electrode (GCE) with a large surface area is prepared to immobilize primary antibodies (Ab1) and provide excellent conductivity...
November 13, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/29287247/aptamer-facilitated-purification-of-functional-proteins
#11
Stanislav S Beloborodov, Jiayin Bao, Svetlana M Krylova, Agnesa Shala-Lawrence, Philip E Johnson, Sergey N Krylov
DNA aptamers are attractive capture probes for affinity chromatography since, in contrast to antibodies, they can be chemically synthesized and, in contrast to tag-specific capture probes (such as Nickel-NTA or Glutathione), they can be used for purification of proteins free of genetic modifications (such as His or GST tags). Despite these attractive features of aptamers as capture probes, there are only a few reports on aptamer-based protein purification and none of them includes a test of the purified protein's activity, thus, leaving discouraging doubts about method's ability to purify proteins in their active state...
December 16, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29277679/intermittent-fasting-promotes-prolonged-associative-interactions-during-synaptic-tagging-capture-by-altering-the-metaplastic-properties-of-the-ca1-hippocampal-neurons
#12
Ananya Dasgupta, Joonki Kim, Anoop Manakkadan, Thiruma V Arumugam, Sreedharan Sajikumar
Metaplasticity is the inherent property of a neuron or neuronal population to undergo activity-dependent changes in neural function that modulate subsequent synaptic plasticity. Here we studied the effect of intermittent fasting (IF) in governing the interactions of associative plasticity mechanisms in the pyramidal neurons of rat hippocampal area CA1. Late long-term potentiation and its associative mechanisms such as synaptic tagging and capture at an interval of 120 min were evaluated in four groups of animals, AL (Ad libitum), IF12 (daily IF for 12 h), IF16 (daily IF for 16 h) and EOD (every other day IF for 24 h)...
December 19, 2017: Neurobiology of Learning and Memory
https://www.readbyqxmd.com/read/29277582/behavioral-tagging-synaptic-event-or-cellular-alteration
#13
REVIEW
Volker Korz
Recently a number of studies were published addressing the translation of the synaptic tagging and capture hypothesis (STCH), developed and tested in hippocampal slices, into learning and long-term memory mechanisms in intact animals. Based on behavioral observations, these studies show similar results as in the original STCH experiments. A weak memory, induced by weak training in one hippocampus dependent behavioral task is converted into a long-term memory, by timely related induction of a long-term memory, induced by strong training in a second hippocampus dependent task...
December 23, 2017: Neurobiology of Learning and Memory
https://www.readbyqxmd.com/read/29241110/design-synthesis-and-initial-evaluation-of-affinity-based-small-molecular-probe-for-detection-of-wdr5
#14
Wei-Lin Chen, Dong-Dong Li, Zhi-Hui Wang, Xiao-Li Xu, Xiao-Jin Zhang, Zheng-Yu Jiang, Xiao-Ke Guo, Qi-Dong You
WDR5, a subunit of the SET/MLL complex, plays critical roles in various biological progresses and are abnormally expressed in many cancers. Here we report the design, synthesis, and biochemical characterization of a new chemical tool to capture WDR5 protein. The probe is a biotinylated version of compound 30 that is a potent WDR5 inhibitor we previously reported. Importantly, the probe displayed high affinity to WDR5 protein in vitro binding potency and showed the ability in specifically and real time monitoring WDR5 protein...
November 28, 2017: Bioorganic Chemistry
https://www.readbyqxmd.com/read/29232830/evidence-of-altered-glycosylation-of-serum-proteins-prior-to-pancreatic-cancer-diagnosis
#15
Shibu Krishnan, Harry J Whitwell, Joy Cuenco, Aleksandra Gentry-Maharaj, Usha Menon, Stephen P Pereira, Marco Gaspari, John F Timms
Biomarkers for the early detection of pancreatic cancer are urgently needed. The aim of this pilot study was to evaluate changes in serum N-glycoproteins and their glycosylation status prior to clinical presentation of pancreatic cancer that may be potential biomarkers. Prediagnosis serum samples pooled according to five time-to-diagnosis groups and a non-cancer control pool were digested with trypsin, labelled with mass tags, and subjected to titanium dioxide capture, deglycosylation, and 2D-LC-MS/MS profiling...
December 9, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29222430/fisheries-independent-surveys-identify-critical-habitats-for-young-scalloped-hammerhead-sharks-sphyrna-lewini-in-the-rewa-delta-fiji
#16
Amandine D Marie, Cara Miller, Celso Cawich, Susanna Piovano, Ciro Rico
Sharp declines in numerous shark populations around the world have generated considerable interest in better understanding and characterising their biology, ecology and critical habitats. The scalloped hammerhead shark (SHS, Sphyrna lewini) is subject to a multitude of natural and anthropogenic threats that are often exacerbated within the coastal embayments and estuaries used during SHS early life stages. In this study, we describe the temporal and spatial distribution, age class composition, and reproductive biology of SHS in the Rewa Delta (RD), Fiji...
December 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29220830/specific-capture-recovery-and-culture-of-cancer-cells-using-oriented-antibody-modified-polystyrene-chips-coated-with-agarose-film
#17
Jiyun Jeong, Yeolin Lee, Yeongeun Yoo, Myung Kyu Lee
Agarose gel can be used for three dimensional (3D) cell culture because it prevents cell attachment. The dried agarose film coated on a culture plate also protected cell attachment and allowed 3D growth of cancer cells. We developed an efficient method for agarose film coating on an oxygen-plasma treated micropost polystyrene chip prepared by an injection molding process. The agarose film was modified to maleimide or Ni-NTA groups for covalent or cleavable attachment of photoactivatable Fc-specific antibody binding proteins (PFcBPs) via their N-terminal cysteine residues or 6xHis tag, respectively...
December 5, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29203204/component-deficits-of-visual-neglect-magnetic-attraction-of-attention-vs-impaired-spatial-working-memory
#18
Monica N Toba, Marco Rabuffetti, Christophe Duret, Pascale Pradat-Diehl, Guido Gainotti, Paolo Bartolomeo
Visual neglect is a disabling consequence of right hemisphere damage, whereby patients fail to detect left-sided objects. Its precise mechanisms are debated, but there is some consensus that distinct component deficits may variously associate and interact in different patients. Here we used a touch-screen based procedure to study two putative component deficits of neglect, rightward "magnetic" attraction of attention and impaired spatial working memory, in a group of 47 right brain-damaged patients, of whom 33 had signs of left neglect...
December 1, 2017: Neuropsychologia
https://www.readbyqxmd.com/read/29192390/epcam-based-assays-for-epithelial-tumor-cell-detection-in-cerebrospinal-fluid
#19
REVIEW
Mark T J van Bussel, Dick Pluim, Mijke Bol, Jos H Beijnen, Jan H M Schellens, Dieta Brandsma
The diagnosis of leptomeningeal metastases (LM) of solid tumors is complicated due to low sensitivities of both magnetic resonance imaging (MRI) and cytology. MRI has a sensitivity of 76% for the diagnosis of LM and cerebrospinal fluid (CSF) cytology has a sensitivity of 44-67% at first lumbar puncture which increases to 84-91% upon second CSF sampling. Epithelial cell adhesion molecule (EpCAM) is expressed by solid tumors of epithelial origin like non-small-cell lung cancer, breast cancer or ovarium cancer...
November 30, 2017: Journal of Neuro-oncology
https://www.readbyqxmd.com/read/29186550/an-objective-neural-signature-of-rapid-perspective-taking
#20
Alexy-Assaf Beck, Bruno Rossion, Dana Samson
The frequency-tagging approach has generally been confined to study low-level sensory processes and always found related activation over the occipital region. Here for the first time, we investigated with it, high-level socio-cognitive functions i.e. the processing of what other people are looking at which is referred to as level 1 visual perspective taking (VPT). Sixteen participants were presented with visual scenes alternating at 2.5 Hz which were depicting a person and an object in a room, while recording electrophysiological brain activity...
November 27, 2017: Social Cognitive and Affective Neuroscience
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