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Miya Kanazawa, Ikiru Atsuta, Yasunori Ayukawa, Takayoshi Yamaza, Ryosuke Kondo, Yuri Matsuura, Kiyoshi Koyano
BACKGROUND: Multipotent mesenchymal stem cells (MSCs) are used clinically in regenerative medicine. Our previous report showed systemically injected MSCs improved peri-implant sealing and accelerated tissue healing. However, the risks of systemic MSC administration, including lung embolism, must be considered; therefore, their local application must be assessed for clinical safety and efficacy. We investigated differences in treatment effect between local and systemic MSC application using a rat oral implantation model...
January 13, 2018: International Journal of Implant Dentistry
Susanita Carvajal, Samantha N Fera, Abby L Jones, Thaisa A Baldo, Islam M Mosa, James F Rusling, Colleen E Krause
Rapidly fabricated, disposable sensor platforms hold tremendous promise for point-of-care detection. Here, we present an inexpensive (< $0.25) fully inkjet printed electrochemical sensor with integrated counter, reference, and working electrodes that is easily scalable for commercial fabrication. The electrochemical sensor platform featured an inkjet printed gold working 8-electrode array (WEA) and counter electrode (CE), along with an inkjet -printed silver electrode that was chlorinated with bleach to produce a Ag/AgCl quasi-reference electrode (RE)...
January 4, 2018: Biosensors & Bioelectronics
Caihong Wang, Jie Gao, Yili Cao, Hongliang Tan
Nanozymes have been extensively exploited in the construction of colorimetric sensors in the past decade. However, as lack of specific recognition element, these sensors are mostly limited in the detection of hydrogen peroxide and its related agents. In this work, a colorimetric assay for the detection of alkaline phosphatase (ALP) activity was developed by employing copper (II)-based metal-organic frameworks (Cu-MOFs) as peroxidase mimic and pyrophosphate (PPi) as recognition element. Benefiting from its intrinsic porosity, Cu-MOFs built from one-step self-assembly showed a superior catalytic activity over horseradish peroxidase...
April 3, 2018: Analytica Chimica Acta
Peter C E Moody, Emma L Raven
Aerobic organisms have evolved to activate oxygen from the atmosphere, which allows them to catalyze the oxidation of different kinds of substrates. This activation of oxygen is achieved by a metal center (usually iron or copper) buried within a metalloprotein. In the case of iron-containing heme enzymes, the activation of oxygen is achieved by formation of transient iron-oxo (ferryl) intermediates; these intermediates are called Compound I and Compound II. The Compound I and II intermediates were first discovered in the 1930s in horseradish peroxidase, and it is now known that these same species are used across the family of heme enzymes, which include all of the peroxidases, the heme catalases, the P450s, cytochrome c oxidase, and NO synthase...
January 12, 2018: Accounts of Chemical Research
Ahamed Khan, Ankita Shrestha, Kashyap Bhuyan, Indu B Maiti, Nrisingha Dey
The promoter fragment described in this study can be employed for strong transgene expression under both biotic and abiotic stress conditions. Plant-infecting Caulimoviruses have evolved multiple regulatory mechanisms to address various environmental stimuli during the course of evolution. One such mechanism involves the retention of discrete stress responsive cis-elements which are required for their survival and host-specificity. Here we describe the characterization of a novel Caulimoviral promoter isolated from Horseradish Latent Virus (HRLV) and its regulation by multiple stress responsive Transcription factors (TFs) namely DREB1, AREB1 and TGA1a...
January 11, 2018: Plant Molecular Biology
Alexander Karich, Sebastian B Kleeberg, René Ullrich, Martin Hofrichter
Enzymatic oxidation of 5-hydroxymethylfurfural (HMF) and its oxidized derivatives was studied using three fungal enzymes: wild-type aryl alcohol oxidase (AAO) from three fungal species, wild-type peroxygenase from Agrocybe aegerita (AaeUPO), and recombinant galactose oxidase (GAO). The effect of pH on different reaction steps was evaluated and apparent kinetic data (Michaelis-Menten constants, turnover numbers, specific constants) were calculated for different enzyme-substrate ratios and enzyme combinations...
January 9, 2018: Microorganisms
Shinichi Sato, Michihiko Tsushima, Kosuke Nakamura, Hiroyuki Nakamura
 The chemical labeling of proteins with synthetic probes is a key technique used in chemical biology, protein-based therapy, and material science. Much of the chemical labeling of native proteins, however, depends on the labeling of lysine and cysteine residues. While those methods have significantly contributed to native protein labeling, alternative methods that can modify different amino acid residues are still required. Herein we report the development of a novel methodology of tyrosine labeling, inspired by the luminol chemiluminescence reaction...
2018: Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan
Xiaoxiao Su, Mengyao Wang, Yue Wu, Yong He, Zhifeng Fu
A novel dual-site recognition protocol was developed for chemiluminescent (CL) detection of Streptococcus mutans (S. mutans) based on a designed antibiotic-affinity strategy. Teicoplanin, a broad-spectrum antibiotic against Gram-positive bacteria, was adopted to functionalize magnetic particles and recognize S. mutans utilizing the strong affinity between this agent and D-Alanyl-D-Alanine peptide moieties in the bacterial cell wall. To achieve ideal specificity for S. mutans detection, rat immunoglobulin G2a (rat IgG2a) tagged with horseradish peroxidase (HRP) was used as the second recognition agent and signal tracer since Fab region of rat IgG2a could bind with streptococcal protein G highly expressed in the cell wall of S...
March 1, 2018: Talanta
Linlin Sun, Qiang Zhao
Aflatoxin B1 (AFB1) is one of highly toxic mycotoxins and a known human carcinogen. The frequent contamination of AFB1 in food products and large health risk of AFB1 have raised global concerns. Sensitive detection of AFB1 is of vital importance and highly demanded. Herein, we reported a competitive horseradish peroxidase (HRP)-linked aptamer assay for AFB1, combining the advantages of aptamer for affinity binding and enzyme label for signal amplification. In this assay, free AFB1 in solution competed with a covalent conjugate of bovine serum albumin-AFB1 (BSA-AFB1) coated on the wells of microplate in binding to the HRP-labeled aptamer probe...
March 1, 2018: Talanta
Se-Ho Kim
Several studies have reported a good correlation between levels of serum hepatitis B virus surface antigen (HBsAg) and covalently closed circular DNA (cccDNA) before and after antiviral therapy. As a result, the quantification of HBsAg levels has attracted much attention in recent years as an important approach to evaluate viral activity. In this study, mAbs against HBsAg were generated and 9 mAbs (H17, H30, H31, H67, H73, H97, H101, H118, and H128) were investigated for optimization of HBsAg quantitation ELISA...
December 2017: Immune Network
Suzanne M Huddy, Inga I Hitzeroth, Ann E Meyers, Brandon Weber, Edward P Rybicki
Horseradish peroxidase (HRP) is a commercially important reagent enzyme used in molecular biology and in the diagnostic product industry. It is typically purified from the roots of the horseradish (Armoracia rusticana); however, this crop is only available seasonally, yields are variable and often low, and the product is a mixture of isoenzymes. Engineering high-level expression in transiently transformed tobacco may offer a solution to these problems. In this study, a synthetic Nicotiana benthamiana codon-adapted full-length HRP isoenzyme gene as well as C-terminally truncated and both N- and C-terminally truncated versions of the HRP C gene were synthesized, and their expression in N...
January 1, 2018: International Journal of Molecular Sciences
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
Jia-Yu Huang, Hong-Ting Lin, Tzu-Heng Chen, Chung-An Chen, Huan-Tsung Chang, Chien-Fu Chen
In this work, we report a highly sensitive colorimetric sensing strategy for cancer biomarker diagnosis using gold nanoparticles (AuNPs) labelled with biotinylated poly-adenine ssDNA sequences and streptavidin-horseradish peroxidase for enzymatic signal enhancement. By adopting this DNA-AuNP nanoconjugate sensing strategy, we were able to eliminate the complicated and costly thiol-binding process typically used to modify AuNP surfaces with ssDNA. In addition, different antibodies can be introduced to the AuNP surfaced via electrostatic interactions to provide highly specific recognition sites for biomolecular sensing...
December 28, 2017: ACS Sensors
Wan-Joong Kim, Hyo Young Cho, Bongjin Jeong, Sangwon Byun, JaeDoo Huh, Young Jun Kim
Using gold nanoparticles (AuNPs) on "capillary enzyme-linked immunosorbent assay (ELISA)", we produced highly sensitive and rapid assays, which are the major attributes for point-of-care applications. First, in order to understand the size effect of AuNPs, AuNPs of varying diameters (5 nm, 10 nm, 15 nm, 20 nm, 30 nm, and 50 nm) conjugated with Horseradish Peroxidase (HRP)-labeled anti-C reactive protein (antiCRP) (AuNP•antiCRP-HRP) were used for well-plate ELISA. AuNP of 10 nm produced the largest optical density, enabling detection of 0...
December 26, 2017: Sensors
Subhash Chand, Sriparna Ray, Eranda Wanigasekara, Poonam Yadav, Joshua A Crawford, Daniel W Armstrong, Krishnan Rajeshwar, Brad S Pierce
This study showcases the potential of unnatural amino acids to enable non-natural functions when incorporated in the protein scaffold of heme metalloproteins. For this purpose, a genetically-engineered myoglobin (Mb) mutant was created by incorporating redox-active 3-amino-l-tyrosine (NH2Tyr) into its active site, replacing the distal histidine (H64) with NH2Tyr. In peroxide-shunt assays, this variant exhibits an increased rate of turnover for thioanisole and benzaldehyde oxidation as compared to the wild-type (WT) Mb...
December 22, 2017: Archives of Biochemistry and Biophysics
Xu Wu, Huanhuan Zhang, Kejing Huang, Yan Zeng, Zhihong Zhu
In this work, highly ordered macro-mesoporous TiO2 has been successfully synthesized using fresh rose petals and P123 (EO20PO70EO20) as dual templates through a simple soaking and calcining process. Characterization of the as-prepared TiO2 indicated that the mesoporous structure of the TiO2 was highly ordered, with a pore diameter of approximately 3nm. After electrodeposition of Pt nanoparticles onto the TiO2 as an electron transfer enhancer and the immobilization of horseradish peroxidase (HRP) onto the TiO2-modified electrode, a biosensor for detecting hydrogen peroxide (H2O2) was realized...
December 19, 2017: Bioelectrochemistry
Jian Sun, Jiahui Zhao, Lei Wang, Hongwei Li, Fan Yang, Xiurong Yang
Being an important model peroxidase, horseradish peroxidase (HRP) has been thoroughly understood, and the detection of HRP is not only directly related to peroxidase-triggered catalytic process, but also linked to the development of HRP-based enzyme-linked immunosorbent assay (ELISA). Herein, we have reported an unconventional fluorescent sensor for convenient assay of HRP activity based on the HRP-catalyzed specific conversion of p-phenylenediamine (PPD) into chromogenic PPDox with H2O2 as the oxidizing agent, accompanied with the fluorescence quenching effect on fluorescein...
December 20, 2017: ACS Sensors
Ming-Ke Tang, Yue-Ming Sun, Li-Juan Qin, Yong-Sen Jia, Tian Zhang, Wei Zhang
OBJECTIVE: To determine the effect of prostaglandin E2 (PGE2) on brain metastasis of lung cancer,and to explore the possible mechanism of aspirin (PGE2 inhibitor) reducing brain metastasis of lung cancer. METHODS: Radioimmunoassay was performed to measure the expression level of PGE2 in cell supernatant collected from cells treated with or without aspirin (8 mmol/L) at different time points. After establishing in vitro blood-brain barrier (BBB) model using Transwell, lung cancer cells was added to upper chamber of transwell and was then treated with aspirin (8 mmol/L)...
November 2017: Sichuan da Xue Xue Bao. Yi Xue Ban, Journal of Sichuan University. Medical Science Edition
Wei Cai, Hui-Hui Li, Zhe-Xue Lu, Maryanne M Collinson
A hierarchical imprinting strategy was used to create protein imprints in a silicate film with a high binding capacity as well as selectivity toward the imprint protein and little specificity towards other proteins. In the first part of this work, rod-shaped bacteria were used as templates to create imprints in silica films of various thicknesses to open up the silica framework and increase the surface area exposed to solution. In the second part, the protein (e.g., cytochrome c (CYC) or green fluorescent protein (GFP)) was covalently attached to the surface of Bacillus subtilis and this protein-bacteria complex served as the imprint moiety...
December 20, 2017: Analyst
Mircea Cătălin ForŢofoiu, Dragoş Marian Popescu, Vlad Pădureanu, Aurelian Constantin Dobrinescu, Adrian Gabriel Dobrinescu, Adrian Mită, Maria Camelia Foarfă, Violeta Serenada Bălă, Anca Emanuela Muşetescu, Nina Ionovici, Maria ForŢofoiu
Ascites is the most frequent complication of cirrhosis and occurs only when the portal hypertension has already installed but ascites is caused by neoplasms, heart failure, tuberculosis, pancreatic illnesses, as well as other kind of affections. We describe the case of a 67-year-old patient, a retired person, without significant personal or familial history, nonsmoker, infrequent alcohol and coffee consumer with following chief complaints at onset: loss of appetite, weight loss, serious physical asthenia, delayed intestinal transit, diffuse abdominal pain and increase of abdominal circumference...
2017: Romanian Journal of Morphology and Embryology, Revue Roumaine de Morphologie et Embryologie
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