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https://www.readbyqxmd.com/read/28097171/structural-insights-into-the-effects-of-charge-reversal-substitutions-at-the-surface-of-horseradish-peroxidase
#1
Leila Navapour, Navid Mogharrab
Horseradish peroxidase (HRP), has gained significant interests in biotechnology, especially in biosensor field and diagnostic test kits. Hence, its solvent-exposed lysine residues 174, 232, and 241 have been frequently modified with the aim of improving its stability and catalytic efficiency. In this computational study, we investigated the effects of Lys-to-Glu substitutions on HRP structure to model charge-reversal manipulations at the enzyme surface. Simulation results implied that upon these substitutions, the number of stable hydrogen bonds and α-helical content of HRP are increased and the proximal Ca2+ binding pocket becomes more integrated...
September 2016: Molecular Biology Research Communications
https://www.readbyqxmd.com/read/28093122/sequential-enzymatic-derivatization-coupled-with-online-microdialysis-sampling-for-simultaneous-profiling-of-mouse-tumor-extracellular-hydrogen-peroxide-lactate-and-glucose
#2
Cheng-Kuan Su, Po-Jen Tseng, Hsien-Ting Chiu, Andrea Del Vall, Yu-Fen Huang, Yuh-Chang Sun
Probing tumor extracellular metabolites is a vitally important issue in current cancer biology. In this study an analytical system was constructed for the in vivo monitoring of mouse tumor extracellular hydrogen peroxide (H2O2), lactate, and glucose by means of microdialysis (MD) sampling and fluorescence determination in conjunction with a smart sequential enzymatic derivatization scheme-involving a loading sequence of fluorogenic reagent/horseradish peroxidase, microdialysate, lactate oxidase, pyruvate, and glucose oxidase-for step-by-step determination of sampled H2O2, lactate, and glucose in mouse tumor microdialysate...
March 1, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28089929/impact-of-immobilizing-of-low-molecular-weight-hyaluronic-acid-within-gelatin-based-hydrogel-through-enzymatic-reaction-on-behavior-of-enclosed-endothelial-cells
#3
Mehdi Khanmohammadi, Shinji Sakai, Masahito Taya
The hydrogels having the ability to promote migration and morphogenesis of endothelial cells (ECs) are useful for fabricating vascularized dense tissues in vitro. The present study explores the immobilization of low molecular weight hyaluronic acid (LMWHA) derivative within gelatin-based hydrogel to stimulate migration of ECs. The LMWHA derivative possessing phenolic hydroxyl moieties (LMWHA-Ph) was bound to gelatin-based derivative hydrogel through the horseradish peroxidase-catalyzed reaction. The motility of ECs was analyzed by scratch migration assay and microparticle-based cell migration assay...
January 12, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28088684/changes-in-electroencephalographic-characteristics-and-blood-brain-barrier-permeability-in-wag-rij-rats-with-cortical-dysplasia
#4
Deniz Sahin, Canan Ugur Yilmaz, Nurcan Orhan, Nadir Arican, Mehmet Kaya, Candan Gürses, Nurbay Ates, Bulent Ahishali
PURPOSE: This study investigated the effects of cortical dysplasia (CD) on electrophysiology and blood-brain barrier (BBB) permeability in WAG/Rij rats with genetic absence epilepsy. METHODS: Pregnant WAG/Rij rats were exposed to 145cGy of gamma-irradiation on embryonic day 17 to induce CD. An electroencephalogram was recorded from cortices subdurally in the offspring of the pregnant animals. Horseradish peroxidase (HRP) was used as determinant of BBB permeability...
January 12, 2017: Epilepsy & Behavior: E&B
https://www.readbyqxmd.com/read/28088341/evaluation-of-an-in-house-indirect-elisa-for-detection-of-antibody-against-haemorrhagic-septicemia-in-asian-elephants
#5
Pallop Tankaew, Thawatchai Singh-La, Chatchote Titaram, Veerasak Punyapornwittaya, Preeyanat Vongchan, Takuo Sawada, Nattawooti Sthitmatee
Pasteurella multocida causes haemorrhagic septicemia in livestock and wild animals, including elephants. The disease has been reported in Asian elephants in India and Sri Lanka, but to date there have been no reported cases in Thailand. ELISA or indirect hemagglutination assays (IHA) have been demonstrated to be able to detect the antibody against the disease in cattle, but no data are available for elephants. The present study reports a novel in-house indirect ELISA for antibody detection of haemorrhagic septicemia in Asian elephants, and evaluates the sensitivity and specificity of the method using a Bayesian approach...
January 12, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/28078652/free-and-ca-alginate-beads-immobilized-horseradish-peroxidase-for-the-removal-of-reactive-dyes-an-experimental-and-modeling-study
#6
Simone Farias, Diego A Mayer, Débora de Oliveira, Selene M A Guelli U de Souza, Antônio Augusto Ulson de Souza
The aim of this work was to remove the dyes Reactive Blue 221 (RB 221) and Reactive Blue 198 (RB 198) of synthetic effluent using the immobilized enzyme horseradish peroxidase (HRP) in Ca-alginate beads. Experimental parameters affecting the dye removal process such as the effect of pH, temperature, hydrogen peroxide concentration, mass capsules, and reuse were evaluated, and a numerical model of mass transfer was developed. A maximum removal of 93 and 75%, respectively, for the dyes RB 221 and RB 198, at pH 5...
January 11, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28072547/mild-photochemical-biofunctionalization-of-glass-microchannels
#7
Rui Rijo Carvalho, Sidharam P Pujari, Digvijay Gahtory, Elwin X Vrouwe, Bauke Albada, Han Zuilhof
The ability to locally modify the inside of microfluidic channels with bioactive molecules is of ever-rising relevance. In this article, we show the direct photochemical coupling of a N-hydroxysuccinimide-terminated ω-alkene onto hydrogen-terminated silicon oxide, and its subsequent functionalization with a catalytically active DNAzyme. To achieve this local attachment of a DNAzyme, we prepared hydrogen-phenyl-terminated glass (H-Φ-glass) by the reaction of glass with H-SiPhCl2. The presence of a radical-stabilizing substituent on the Si atom (i...
January 10, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28070623/synaptic-contact-between-median-preoptic-neurons-and-subfornical-organ-neurons-projecting-to-the-paraventricular-hypothalamic-nucleus
#8
Hitoshi Kawano
It is known that the median preoptic nucleus (POMe) sends dense projections to the subfornical organ (SFO). However, the functional significance of these projections have not been well discussed. In this electron microscopic study, we investigated the presence of synapses between POMe-derived axon terminals and SFO neurons that project to the paraventricular hypothalamic nucleus (PVN). After injection of a retrograde tracer, wheat germ agglutinin-conjugated horseradish peroxidase-colloidal gold complex, into the PVN, many labeled neurons were found in the SFO...
January 9, 2017: Experimental Brain Research. Experimentelle Hirnforschung. Expérimentation Cérébrale
https://www.readbyqxmd.com/read/28068756/hydrophilic-mesoporous-silica-materials-for-highly-specific-enrichment-of-n-linked-glycopeptide
#9
Nianrong Sun, Jiawen Wang, Jizong Yao, Chun-Hui Deng
Hydrophilic interaction liquid chromatography (HILIC) is a significant enrichment strategy in glycoproteomics profiling. In this report, hydrophilic magnetic mesoporous silica materials (Fe3O4@mSiO2-IDA) were designed and synthesized as an outstanding enrichment platform for glycopeptide analysis. By taking advantage of their merits, such as large surface area, excellent hydrophilicity and unbiased affinity toward all types of glycopeptides, the Fe3O4@mSiO2-IDA nanomaterials were successfully applied to capture glycopeptides from complex samples...
January 4, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28068469/porphyrin-based-porous-organic-frameworks-as-a-biomimetic-catalyst-for-highly-efficient-colorimetric-immunoassay
#10
Xiao Deng, Yishan Fang, Sha Lin, Qian Cheng, Qingyun Liu, Xiaomei Zhang
Herein, we synthesized a cost-effective iron porphyrin (FePor)-based covalent organic polymer (COP), FePor-TFPA-COP, through an easy aromatic substitution reaction between pyrrole and tris(4-formylphenyl)amine (TFPA). The triangular pyramid-shaped, N-centric structure of TFPA facilitated the formation of FePor-TFPA-COP with three-dimensional porous structure, larger surface area, and abundant surface catalytically active sites. FePor-TFPA-COP exhibited strong intrinsic peroxidase activity toward a classical peroxidase substrate, 3,3',5,5'-tetramethylbenzidine (TMB), in the presence of H2O2...
January 18, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28067526/extraction-of-thermodynamic-parameters-of-protein-unfolding-using-parallelized-differential-scanning-fluorimetry
#11
Thaiesha A Wright, Jamie M Stewart, Richard Christopher Page, Dominik Konkolewicz
Thermodynamic properties of protein unfolding have been extensively studied; however, the methods used have typically required significant preparation time and high protein concentrations. Here, we present a facile, simple, and parallelized differential scanning fluorimetry (DSF) method that enables thermodynamic parameters of protein unfolding to be extracted. This method assumes a two-state, reversible protein unfolding mechanism and provides the capacity to quickly analyze the biophysical mechanisms of changes in protein stability and to more thoroughly characterize the effect of mutations, additives, inhibitors, or pH...
January 9, 2017: Journal of Physical Chemistry Letters
https://www.readbyqxmd.com/read/28067370/automated-3d-printed-unibody-immunoarray-for-chemiluminescence-detection-of-cancer-biomarker-proteins
#12
C K Tang, A Vaze, J F Rusling
A low cost three-dimensional (3D) printed clear plastic microfluidic device was fabricated for fast, low cost automated protein detection. The unibody device features three reagent reservoirs, an efficient 3D network for passive mixing, and an optically transparent detection chamber housing a glass capture antibody array for measuring chemiluminescence output with a CCD camera. Sandwich type assays were built onto the glass arrays using a multi-labeled detection antibody-polyHRP (HRP = horseradish peroxidase)...
January 9, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28058577/development-of-an-indirect-dot-ppa-elisa-using-glutamate-dehydrogenase-as-a-diagnostic-antigen-for-the-rapid-and-specific-detection-of-streptococcus-suis-and-its-application-to-clinical-specimens
#13
Xiao-Jing Xia, Lei Wang, Zhi-Qiang Shen, Wanhai Qin, Jianhe Hu, Shi-Jin Jiang, Shu-Guang Li
Streptococcus suis is an important zoonotic pathogen causing infections in pigs and humans. Bacterial surface-related proteins are often explored as potential vaccine candidates and diagnostic antigens. In the present study, glutamate dehydrogenase, a highly conserved immunogenic extracellular protein, was used to establish a dot horseradish peroxidase enzyme-linked staphylococcal protein A immunosorbent assay (Dot-PPA-ELISA) for diagnosis of S. suis infection. The antigen-antibody reaction was optimised through checkerboard titration involving serial dilutions, followed by selective blocking tests and evaluations of cross-reaction, repeatability, and stability...
January 5, 2017: Antonie Van Leeuwenhoek
https://www.readbyqxmd.com/read/28057474/hiv-1-tat-disrupts-blood-brain-barrier-integrity-and-increases-phagocytic-perivascular-macrophages-and-microglia-in-the-dorsal-striatum-of-transgenic-mice
#14
Crystal R Leibrand, Jason J Paris, M Said Ghandour, Pamela E Knapp, Woong-Ki Kim, Kurt F Hauser, MaryPeace McRae
HIV-1 infection results in blood brain barrier (BBB) disruption, which acts as a rate-limiting step for HIV-1 entry into the CNS and for subsequent neuroinflammatory/neurotoxic actions. One mechanism by which HIV may destabilize the BBB involves actions of the HIV-1 regulatory protein, trans-activator of transcription (Tat). We utilized a conditional, Tat-expressing transgenic murine model to examine the influence of Tat1-86 expression on BBB integrity and to assess the relative numbers of phagocytic perivascular macrophages and microglia within the CNS in vivo...
January 2, 2017: Neuroscience Letters
https://www.readbyqxmd.com/read/28049107/electrochemical-sensor-based-on-magnetic-molecularly-imprinted-nanoparticles-modified-magnetic-electrode-for-determination-of-hb
#15
Binghua Sun, Xinjiong Ni, Yuhua Cao, Guangqun Cao
A fast and selective electrochemical sensor for determination of hemoglobin (Hb) was developed based on magnetic molecularly imprinted nanoparticles modified on the magnetic glassy carbon electrode. The nanoparticles Fe3O4@SiO2 with a magnetic core and a molecularly imprinted shell had regular structures and good monodispersity. Hb could be determined directly by electrochemical oxidization with the modified electrode. A magnetic field increased electrochemical response to Hb by two times. Imprinting Hb on the surface of Fe3O4@SiO2 shortened the response time within 7min...
December 28, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28043862/rapid-and-facile-quantitation-of-polyplex-endocytic-trafficking
#16
Mihael Lazebnik, Daniel W Pack
Design of safe and effective synthetic nucleic acid delivery vectors such as polycation/DNA or polycation/siRNA complexes (polyplexes) will be facilitated by quantitative understanding of the mechanisms by which such materials escort cargo from the cell surface to the nucleus. In particular, the mechanisms of cellular internalization by various endocytosis pathways and subsequent endocytic vesicle trafficking have been shown to strongly affect nucleic acid delivery efficiency. Fluorescence microscopy and subcellular fractionation methods are commonly employed to follow intracellular trafficking of biomolecules and nanoparticulate delivery systems such as polyplexes...
December 31, 2016: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28035806/microfluidic-reactors-based-on-rechargeable-catalytic-porous-supports-heterogeneous-enzymatic-catalysis-via-reversible-host-guest-interactions
#17
Alberto Sanz de León, Nelson Vargas-Alfredo, Alberto Gallardo, Alfonso Fernández-Mayoralas, Agatha Bastida, Alexandra Muñoz-Bonilla, Juan Rodríguez-Hernández
We report on the fabrication of a microfluidic device in which the reservoir contains a porous surface with enzymatic catalytic activity provided by the reversible immobilization of horseradish peroxidase onto micrometer size pores. The porous functional reservoir was obtained by the Breath Figures approach by casting in a moist environment a solution containing a mixture of high molecular weight polystyrene (HPS) and a poly(styrene-co-cyclodextrin based styrene) (P(S-co-SCD)) statistical copolymer. The pores enriched in CD were employed to immobilize horseradish peroxidase (previously modified with adamantane) by host-guest interactions (HRP-Ada)...
January 17, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28030766/boosted-sensor-performance-by-surface-modification-of-bifunctional-rht-type-metal-organic-framework-with-nanosized-electrochemically-reduced-graphene-oxide
#18
Cong Li, Tingting Zhang, Jingyu Zhao, He Liu, Bo Zheng, Yue Gu, Xiaoyi Yan, Yaru Li, Nannan Lu, Zhiquan Zhang, Guodong Feng
The surface and interface could be designed to enhance properties of electrocatalysts, and they are regarded as the key characteristics. This report describes surface modification of a bifunctional rht-type metal-organic framework (MOF, Cu-TDPAT) with nanosized electrochemically reduced graphene oxide (n-ERGO). The hybrid strategy results in a Cu-TDPAT-n-ERGO sensor with sensitive and selective response toward hydrogen peroxide (H2O2). Compared with Cu-TDPAT, Cu-TDPAT-n-ERGO exhibits significantly enhanced electrocatalytic activities, highlighting the importance of n-ERGO in boosting their electrocatalytic activity...
January 10, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28025846/prolactin-and-growth-hormone-immunoactivity-in-canine-mammary-adenomas-and-adenocarcinomas
#19
E R Bohrer, C V Löhr, M A Kutzler
It is now widely accepted in human medicine that prolactin (PRL) and growth hormone (GH) function in the mammary gland in an autocrine and paracrine manner in tumour formation. The aim of this study was to compare PRL and GH immunoactivity in canine mammary tumours submitted for histopathologic evaluation. Formalin-fixed specimens from spontaneously occurring mammary adenomas and adenocarcinomas from 24 female client-owned dogs were used. Information pertaining to the reproductive status of the patient at the time of mammary tumour diagnosis was obtained from each of the submitting veterinarians...
December 27, 2016: Reproduction in Domestic Animals, Zuchthygiene
https://www.readbyqxmd.com/read/28024284/chemiluminescence-cloth-based-glucose-test-sensors-ccgtss-a-new-class-of-chemiluminescence-glucose-sensors
#20
Huijie Li, Cuiling Liu, Dan Wang, Chunsun Zhang
Chemiluminescence (CL) has been widely applied in many fields, but it is rarely used in a very simple, economical but effective way. In this work, for the first time, the CL cloth-based glucose test sensors (CCGTSs) are developed as a new class of CL glucose sensors, with no need for complicated, expensive device fabrication and peripheral equipment. When integrated with desirable hydrophobic barrier in the flow channel and gravity/capillary flow induced by a difference in height between the loading zone and the detection zone, a single cloth-based device can perform the whole CL process involving two steps of enzyme reactions...
December 2, 2016: Biosensors & Bioelectronics
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