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https://www.readbyqxmd.com/read/28727490/comparison-of-the-release-of-micrornas-and-extracellular-vesicles-from-platelets-in-response-to-different-agonists
#1
Ashley R Ambrose, Mohammed A Alsahli, Sameer A Kurmani, Alison H Goodall
On activation platelets release microRNAs and extracellular vesicles (EV) into circulation. The release of EV from platelets has been shown to be dependent on the agonist; in this study, we investigated whether the microRNA profile or EV released from platelets was also agonist specific. Washed platelets from healthy subjects were maximally stimulated with agonists specific for the receptors for collagen (Glycoprotein VI (GPVI)), thrombin (PAR1/PAR4), or ADP (P2Y1/P2Y12) with/without inhibiting secondary mediators, using aspirin to block cyclooxygenase-1 and apyrase to remove ADP...
July 20, 2017: Platelets
https://www.readbyqxmd.com/read/28724621/mildew-resistance-locus-function-in-pollen-ovary-reception-is-linked-to-its-oligomerization-and-subcellular
#2
Daniel S Jones, Jing Yuan, Benjamin Smith, Andrew Willoughby, Emily L Kumimoto, Sharon A Kessler
Sexual reproduction in flowering plants requires communication between synergid cells and a tip-elongating pollen tube (PT) for the successful delivery of sperm cells to the embryo sac. The reception of the PT relies on signaling within the synergid cell that ultimately leads to the degeneration of the receptive synergid and PT rupture, releasing the sperm cells for double fertilization. In Arabidopsis thaliana, NORTIA (NTA), a member of the Mildew resistance locus O (MLO) family of proteins, plays a critical role in the communication processes regulating PT reception...
July 19, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28717425/high-throughput-nanoparticle-tracking-analysis-for-monitoring-outer-membrane-vesicle-production
#3
Matthias J H Gerritzen, Dirk E Martens, René H Wijffels, Michiel Stork
Outer membrane vesicles (OMVs) are spherical membrane nanoparticles released by Gram-negative bacteria. OMVs can be quantified in complex matrices by nanoparticle tracking analysis (NTA). NTA can be performed in static mode or with continuous sample flow that results in analysis of more particles in a smaller time-frame. Flow measurements must be performed manually despite the availability of a sample changer on the NanoSight system. Here we present a method for automated measurements in flow mode. OMV quantification in flow mode results in lower variance in particle quantification (coefficient of variation (CV) of 6%, CV static measurements of 14%)...
2017: Journal of Extracellular Vesicles
https://www.readbyqxmd.com/read/28717417/analysis-of-extracellular-rna-in-cerebrospinal-fluid
#4
Julie A Saugstad, Theresa A Lusardi, Kendall R Van Keuren-Jensen, Jay I Phillips, Babett Lind, Christina A Harrington, Trevor J McFarland, Amanda L Courtright, Rebecca A Reiman, Ashish S Yeri, M Yashar S Kalani, P David Adelson, Jorge Arango, John P Nolan, Erika Duggan, Karen Messer, Johnny C Akers, Douglas R Galasko, Joseph F Quinn, Bob S Carter, Fred H Hochberg
We examined the extracellular vesicle (EV) and RNA composition of pooled normal cerebrospinal fluid (CSF) samples and CSF from five major neurological disorders: Alzheimer's disease (AD), Parkinson's disease (PD), low-grade glioma (LGG), glioblastoma multiforme (GBM), and subarachnoid haemorrhage (SAH), representing neurodegenerative disease, cancer, and severe acute brain injury. We evaluated: (I) size and quantity of EVs by nanoparticle tracking analysis (NTA) and vesicle flow cytometry (VFC), (II) RNA yield and purity using four RNA isolation kits, (III) replication of RNA yields within and between laboratories, and (IV) composition of total and EV RNAs by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and RNA sequencing (RNASeq)...
2017: Journal of Extracellular Vesicles
https://www.readbyqxmd.com/read/28712956/refolding-of-a-novel-cholesterol-oxidase-from-pimelobacter-simplex-reveals-dehydrogenation-activity
#5
Hui-Min Qin, Jian-Wen Wang, Qianqian Guo, Songtao Li, Panpan Xu, Zhangliang Zhu, Dengyue Sun, Fuping Lu
Cholesterol oxidases, which catalyze the degradation of cholesterol to cholest-4-en-3-one, are widely used in the pharmaceutical and food processing industries. The cholesterol oxidase from Pimelobacter simplex (PsChO3) was transformed into E. coli BL21(DE3), but it was expressed mainly as inclusion bodies, and any soluble PsChO3 failed to bind to Ni-NTA resin. To overcome this obstacle, we devised a simple yet efficient purification and refolding process using 8 M urea for the solubilization of PsChO3 and achieved a high yield of the enzyme in its active form...
July 13, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28706572/a-microwell-printing-fabrication-strategy-for-the-on-chip-templated-biosynthesis-of-protein-microarrays-for-surface-plasmon-resonance-imaging
#6
Gerald Manuel, Andrej Lupták, Robert M Corn
A two-step templated, ribosomal biosynthesis/printing method for the fabrication of protein microarrays for surface plasmon resonance imaging (SPRI) measurements is demonstrated. In the first step, a sixteen component microarray of proteins is created in microwells by cell free on chip protein synthesis; each microwell contains both an in vitro transcription and translation (IVTT) solution and 350 femtomoles of a specific DNA template sequence that together are used to create approximately 40 picomoles of a specific hexahistidine-tagged protein...
September 22, 2016: Journal of Physical Chemistry. C, Nanomaterials and Interfaces
https://www.readbyqxmd.com/read/28706308/introduction-to-nematode-genome-and-transcriptome-announcements-in-the-journal-of-nematology
#7
Dee R Denver, Erik J Ragsdale, W Kelley Thomas, Inga A Zasada
The Journal of Nematology now offers publication of Nematode Genome Announcements (NGA) and Nematode Transcriptome Announcements (NTA). These brief reports announce the sequencing and assembly of a nematode genome or transcriptome resource, along with basic technical information on DNA sequencing and bioinformatic methods used. This publishing initiative offers a new avenue to openly and concisely communicate the availability and relevance of genome and transcriptome sequence resources to the broader scientific community...
June 2017: Journal of Nematology
https://www.readbyqxmd.com/read/28706267/a-simple-add-and-display-method-for-immobilisation-of-cancer-drug-on-his-tagged-virus-like-nanoparticles-for-controlled-drug-delivery
#8
Roya Biabanikhankahdani, Saadi Bayat, Kok Lian Ho, Noorjahan Banu Mohamed Alitheen, Wen Siang Tan
pH-responsive virus-like nanoparticles (VLNPs) hold promising potential as drug delivery systems for cancer therapy. In the present study, hepatitis B virus (HBV) VLNPs harbouring His-tags were used to display doxorubicin (DOX) via nitrilotriacetic acid (NTA) conjugation. The His-tags served as pH-responsive nanojoints which released DOX from VLNPs in a controlled manner. The His-tagged VLNPs conjugated non-covalently with NTA-DOX, and cross-linked with folic acid (FA) were able to specifically target and deliver the DOX into ovarian cancer cells via folate receptor (FR)-mediated endocytosis...
July 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28698826/expression-refolding-and-spectroscopic-characterization-of-fibronectin-type-iii-fniii-homology-domains-derived-from-human-fibronectin-leucine-rich-transmembrane-protein-flrt-1-2-and-3
#9
Lila Yang, Maria Hansen Falkesgaard, Peter Waaben Thulstrup, Peter Schledermann Walmod, Leila Lo Leggio, Kim Krighaar Rasmussen
The fibronectin leucine rich transmembrane (FLRT) protein family consists in humans of 3 proteins, FLRT1, -2, and -3. The FLRT proteins contain two extracellular domains separated by an unstructured linker. The most membrane distal part is a leucine rich repeat (LRR) domain responsible for both cis- and trans-interactions, whereas the membrane proximal part is a fibronectin type III (FnIII) domain responsible for a cis-interaction with members of the fibroblast growth factor receptor 1 (FGFR1) family, which results in FGFR tyrosine kinase activation...
2017: PeerJ
https://www.readbyqxmd.com/read/28694142/increased-psa-expression-on-prostate-cancer-exosomes-in-in%C3%A2-vitro-condition-and-in-cancer-patients
#10
Mariantonia Logozzi, Daniela F Angelini, Elisabetta Iessi, Davide Mizzoni, Rossella Di Raimo, Cristina Federici, Luana Lugini, Giovanna Borsellino, Alessandro Gentilucci, Federico Pierella, Vittorio Marzio, Alessandro Sciarra, Luca Battistini, Stefano Fais
Prostate specific antigen (PSA) test is the most common, clinically validated test for the diagnosis of prostate cancer (PCa). While neoplastic lesions of the prostate may cause aberrant levels of PSA in the blood, the quantitation of free or complexed PSA poorly discriminates cancer patients from those developing benign lesions, often leading to invasive and unnecessary surgical procedures. Microenvironmental acidity increases exosome release by cancer cells. In this study we evaluated whether acidity, a critical phenotype of malignancy, could influence exosome release and increase the PSA expression in nanovesicles released by PCa cells...
July 8, 2017: Cancer Letters
https://www.readbyqxmd.com/read/28684315/efficient-renaturation-of-inclusion-body-proteins-denatured-by-sds
#11
Chuan He, Kouhei Ohnishi
Inclusion bodies are often formed when the foreign protein is over expressed in Escherichia coli. Since proteins in inclusion bodies are inactive, denaturing and refolding of inclusion body proteins are necessary to obtain the active form. Instead of the conventional denaturants, urea and guanidine hydrochloride, a strong anionic detergent SDS was used to solubilize C-terminal His-tag form of ulvan lyase in the inclusion bodies. Solution containing SDS-solubilized enzyme were kept on ice to precipitate SDS, followed by SDS-KCl insoluble crystal formation to remove SDS completely...
July 3, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28684274/construction-of-high-level-prokaryotic-expression-and-purification-system-of-pd-l1-extracellular-domain-by-using-escherichia-coli-host-cell-machinery
#12
Muhammad Kalim, Jie Chen, Shenghao Wang, Caiyao Lin, Saif Ullah, Keying Liang, Qian Ding, Shuqing Chen, Jinbiao Zhan
Programmed cell death 1 ligand 1 (PD-L1) is a trans-membrane protein highly expressed on the membrane of cancer cell, which binds inhibitory receptor of PD-1 on the T cells and attenuates anti-tumor immune response.The strategy of blocking PD1 and PD-L1 interaction has been widely used for anti-cancer drug development. The DNA encoding extracellular domain of PD-L1wascloned and expressed with the pET30(+) and Escherichia coli BL21(DE3) system. Cloning of PD-L1 extracellular domain was confirmed by PCR and enzymatic digestion...
July 3, 2017: Immunology Letters
https://www.readbyqxmd.com/read/28675620/site-specific-sers-assay-for-survivin-protein-dimer-from-ensemble-experiments-to-correlative-single-particle-imaging
#13
Jörg Wissler, Sandra Bäcker, Alessandro Feis, Shirley K Knauer, Sebastian Schlücker
An assay for Survivin, a small dimeric protein which functions as modulator of apoptosis and cell division and serves as a promising diagnostic biomarker for different types of cancer, is presented. The assay is based on switching on surface-enhanced Raman scattering (SERS) upon incubation of the Survivin protein dimer with Raman reporter-labeled gold nanoparticles (AuNP). Site-specificity is achieved by complexation of nickel-chelated N-nitrilo-triacetic acid (Ni-NTA) anchors on the particle surface by multiple histidines (His6 -tag) attached to each C-terminus of the centrosymmetric protein dimer...
July 4, 2017: Small
https://www.readbyqxmd.com/read/28668496/generation-of-nanobodies-against-slyd-and-development-of-tools-to-eliminate-this-bacterial-contaminant-from-recombinant-proteins
#14
Yaozhong Hu, Ema Romão, Didier Vertommen, Cécile Vincke, Francisco Morales-Yánez, Carlos Gutiérrez, Changxiao Liu, Serge Muyldermans
The gene for a protein domain, derived from a tumor marker, fused to His tag codons and under control of a T7 promotor was expressed in E. coli strain BL21 (DE3). The recombinant protein was purified from cell lysates through immobilized metal affinity chromatography and size-exclusion chromatography. A contaminating bacterial protein was consistently co-purified, even using stringent washing solutions containing 50 or 100 mM imidazole. Immunization of a dromedary with this contaminated protein preparation, and the subsequent generation and panning of the immune Nanobody library yielded several Nanobodies of which 2/3 were directed against the bacterial contaminant, reflecting the immunodominance of this protein to steer the dromedary immune response...
June 28, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28667645/recombinant-production-of-aspergillus-flavus-uricase-and-investigation-of-its-thermal-stability-in-the-presence-of-raffinose-and-lactose
#15
Mehdi Imani, Serveh Shahmohamadnejad
Aspergillus flavus uricase (Rasburicase) with a molecular mass of 135 kDa is currently used for the treatment of gout and hyperuricemia occurring in tumor lysis syndrome. To characterize the effects of raffinose and lactose osmolytes on the uricase structure, its coding sequence was cloned, expressed in E. coli BL21, and purified by Ni-NTA agarose affinity chromatography. Thermal inactivation studies at 40 °C showed that nearly 15% of UOX activity was preserved, while the presence of raffinose and lactose reduced its activity to 35 and 45% of its original activity, respectively...
July 2017: 3 Biotech
https://www.readbyqxmd.com/read/28664493/assessment-of-pb-and-pyrene-accumulation-in-scirpus-triqueter-assisted-by-combined-alkyl-polyglucoside-and-nitrilotriacetic-acid-application
#16
Tingru Chen, Xiaoyan Liu, Xinying Zhang, Xiaoxin Hu, Liya Cao
To understand the accumulation and uptake of polycyclic aromatic hydrocarbons (PAHs) and heavy metals by plants is an important part of the assessment of phytoremediation for PAHs and heavy metals co-contaminated soil. This study was an investigation of the accumulation and uptake of pyrene and lead (Pb) by Scirpus triqueter under the condition of alkyl polyglucoside (APG) and nitrilotriacetic acid (NTA) combined application. The results indicated that the accumulation of Pb by S. triqueter was significantly improved by NTA and APG addition into the soil...
June 29, 2017: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/28646396/enhanced-phytoextraction-of-lead-from-artificially-contaminated-soil-by-mirabilis-jalapa-with-chelating-agents
#17
Li Yan, Cuilan Li, Jinjing Zhang, Odeshnee Moodley, Shuoran Liu, Chang Lan, Qiang Gao, Wenjing Zhang
A microcosm experiment was conducted to compare how different chelating agents enhance the phytoextraction of lead (Pb) by Mirabilis jalapa from an artificially Pb-contaminated soil. Chelating agents used included ethylene diamine tetraacetic acid (EDTA), nitrilotriacetic acid (NTA), ethylene diamine disuccinic acid (EDDS), oxalic acid, humic acid, citric acid and tartaric acid, with concentrations ranging from 0 to 2000 mg/kg. Results show that with increasing concentrations of chelating agents, the concentrations, bioaccumulation coefficients, and translocation factors of Pb in M...
June 23, 2017: Bulletin of Environmental Contamination and Toxicology
https://www.readbyqxmd.com/read/28644041/identifying-the-functional-groups-and-the-influence-of-synthetic-chelators-on-cd-availability-and-microbial-biomass-carbon-in-cd-contaminated-soil
#18
Muhammad Afzal Chhajro, Qingling Fu, Muhammad Shaaban, Muhammad Shahid Rizwan, Jun Zhu, Abdus Salam, Kashif Ali Kubar, Saqib Bashir, Hongqing Hu, Ghulam Murtaza Jamro
Synthetic chelators play an important role in boosting the microbial biomass carbon (MBC), dissolved organic carbon (DOC) and heavy metal solubility in a contaminated soil towards a sustainability of environment for agricultural crops. Castor was selected as a potential cadmium (Cd) accumulator in different levels of Cd (-Cd and +Cd) contamination with using three chelating agents, ethylenediamine tetraacetic acid (H4EDTA), nitriloacetic acid (H3 NTA) and NH4 citrate (ammonium citrate) were added to soil at rates of 10, 15 and 25 mmol in 5 kg of soil per pot...
June 23, 2017: International Journal of Phytoremediation
https://www.readbyqxmd.com/read/28624685/bacillus-subtilis-affects-mirnas-and-flavanoids-production-in-agrobacterium-tobacco-interaction
#19
Fahimeh Nazari, Naser Safaie, Bahram Mohammad Soltani, Masoud Shams-Bakhsh, Mohsen Sharifi
Agrobacterium tumefaciens is a very destructive plant pathogen. Selection of effective biological agents against this pathogen depends on more insight into molecular plant defence responses during the biocontrol agent-pathogen interaction. Auxin as a phytohormone is a key contributor in pathogenesis and plant defence and accumulation of auxin transport carriers are accompanied by increasing in flavonoid and miRNAs concentrations during plant interactions with bacteria. The aim of this research was molecular analysis of Bacillus subtilis (ATCC21332) biocontrol effect against A...
June 10, 2017: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/28623492/recombinant-20-8-kda-protein-of-mycobacterium-avium-subsp-paratuberculosis-based-sero-diagnosis-of-paratuberculosis
#20
P P Goswami, Gokul Chand, N S Prasad, R Deb, S H Basagoudanavar
Johne's disease or paratuberculosis is a chronic infectious enteric disease of ruminants caused by the intracellular pathogen. The control of the Johne's disease is hampered by lack of specific diagnostic tests. In this study, we have cloned and expressed the N-terminal region of the locus tag Map 1637c encoding 20.8-kDa (r20.8) protein of Mycobacterium avium subsp. paratuberculosis. The recombinant protein r20.8 was expressed in high levels in Escherichia coli. The protein r20.8 was purified by single-step chromatography using Ni-NTA agarose...
July 2017: 3 Biotech
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