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https://www.readbyqxmd.com/read/29783093/a-direct-plasma-mirna-assay-for-early-detection-and-histological-classification-of-lung-cancer
#1
Qixin Leng, Yue Wang, Feng Jiang
Cell-free microRNAs in plasma provide circulating biomarkers for lung cancer. Most techniques for analysis of miRNAs require a large plasma volume to purify a sufficient RNA yield followed by complicated downstream processing. Small differences in the multiple procedures often cause large analytical variations and poor diagnostic values of the plasma biomarkers. Here we investigate whether directly quantifying plasma miRNAs without RNA purification could diagnose lung cancer. FirePlex assay was directly applied to 20 μl plasma of 56 lung cancer patients and 28 cancer free controls for quantifying 11 lung tumor-associated miRNAs...
May 17, 2018: Translational Oncology
https://www.readbyqxmd.com/read/29782982/process-optimization-purification-and-characterization-of-a-novel-acidic-thermostable-chitinase-from-humicola-grisea
#2
Manish Kumar, Amandeep Brar, V Vivekanand, Nidhi Pareek
An extracellular acidic and thermostable chitinase (HgChi) from thermophilic Humicola grisea was purified and characterized. Enhancement in chitinase production (Qp  = 2.9662 Ul-1  h-1 ) was achieved through derivation of optimum fermentation conditions via central composite design. H. grisea observed to produce various isoforms of chitinase, among which the major expressed form has molecular mass of about 50 kDa. Purified chitinases exhibited optimal activity at pH 3.0 and 70 °C. Chitinase showed notable stability at increasing temperatures...
May 18, 2018: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29782014/identification-of-cyclin-dependent-kinase-1-specific-phosphorylation-sites-by-an-in-vitro-kinase-assay
#3
Heying Cui, Kyle M Loftus, Crystal R Noell, Sozanne R Solmaz
Cyclin-dependent kinase 1 (Cdk1) is a master controller for the cell cycle in all eukaryotes and phosphorylates an estimated 8 - 13% of the proteome; however, the number of identified targets for Cdk1, particularly in human cells is still low. The identification of Cdk1-specific phosphorylation sites is important, as they provide mechanistic insights into how Cdk1 controls the cell cycle. Cell cycle regulation is critical for faithful chromosome segregation, and defects in this complicated process lead to chromosomal aberrations and cancer...
May 3, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29782011/a-preclinical-mouse-model-of-osteosarcoma-to-define-the-extracellular-vesicle-mediated-communication-between-tumor-and-mesenchymal-stem-cells
#4
Tonny Lagerweij, Maria Pérez-Lanzón, S Rubina Baglio
Within the tumor microenvironment, resident or recruited mesenchymal stem cells (MSCs) contribute to malignant progression in multiple cancer types. Under the influence of specific environmental signals, these adult stem cells can release paracrine mediators leading to accelerated tumor growth and metastasis. Defining the crosstalk between tumor and MSCs is of primary importance to understand the mechanisms underlying cancer progression and identify novel targets for therapeutic intervention. Cancer cells produce high amounts of extracellular vesicles (EVs), which can profoundly affect the behavior of target cells in the tumor microenvironment or at distant sites...
May 6, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29781987/retrospective-microrna-sequencing-complementary-dna-library-preparation-protocol-using-formalin-fixed-paraffin-embedded-rna-specimens
#5
Olivier Loudig, Christina Liu, Thomas Rohan, Iddo Z Ben-Dov
-Archived, clinically classified formalin-fixed paraffin-embedded (FFPE) tissues can provide nucleic acids for retrospective molecular studies of cancer development. By using non-invasive or pre-malignant lesions from patients who later develop invasive disease, gene expression analyses may help identify early molecular alterations that predispose to cancer risk. It has been well described that nucleic acids recovered from FFPE tissues have undergone severe physical damage and chemical modifications, which make their analysis difficult and generally requires adapted assays...
May 5, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29781870/pregnancy-and-the-apoptotic-pathway-in-experimental-melanoma
#6
Anamaria S Facina, Gil Facina, Ismael D C Guerreiro da Silva, Silvana A A Corrêa, Sandra M Alexandre, Ângela F Logullo, Jorge K Hosomi, Mary U Nakamura
Pregnancy-associated melanoma is defined as melanoma diagnosed during pregnancy or within 1 year of delivery. The association of pregnancy with melanoma is well known, but its underlying molecular mechanisms of association are poorly understood. The aim was to assess the expression of apoptosis-related genes in melanoma tumors during pregnancy in an attempt to elucidate the molecular mechanisms underlying apoptosis-driven activation of melanoma cells in this period. Mice were allocated across two experimental groups (nonpregnant and pregnant) and implanted with the melanoma cell line BF16-F10...
May 17, 2018: Melanoma Research
https://www.readbyqxmd.com/read/29781060/effects-of-physical-and-chemical-aspects-on-membrane-fouling-and-cleaning-using-interfacial-free-energy-analysis-in-forward-osmosis
#7
Wanzhu Zhang, Bingzhi Dong
Natural organic matter (NOM) in micro-polluted water purification using membranes is a critical issue to handle. Understanding the fouling mechanism in the forward osmosis (FO) process, particularly identifying the predominant factor that controls membrane fouling, could have significant effects on exerting the advantages of FO technique. Cellulose triacetate no-woven (CTA-NW) membrane is applied to experiments with a high removal efficiency (> 99%) for the model foulant. Tannic acid (TA) is used as a surrogate foulant for NOM in the membrane fouling process, thus enabling the analysis of the effects of physical and chemical aspects of water flux, retention, and adsorption...
May 20, 2018: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/29781012/microfluidic-curved-channel-centrifuge-for-solution-exchange-of-target-microparticles-and-their-simultaneous-separation-from-bacteria
#8
Pouriya Bayat, Pouya Rezai
One of the common operations in sample preparation is to separate specific particles (e.g. target cells, embryos or microparticles) from non-target substances (e.g. bacteria) in a fluid and to wash them into clean buffers for further processing like detection (called solution exchange in this paper). For instance, solution exchange is widely needed in preparing fluidic samples for biosensing at the point-of-care and point-of-use, but still conducted via the use of cumbersome and time-consuming off-chip analyte washing and purification techniques...
May 21, 2018: Soft Matter
https://www.readbyqxmd.com/read/29780475/development-of-microfluidic-platforms-for-the-synthesis-of-metal-complexes-and-evaluation-of-their-dna-affinity-using-online-fret-melting-assays
#9
Viktoria Rakers, Paolo Cadinu, Joshua B Edel, Ramon Vilar
Guanine-rich DNA sequences can fold into quadruple-stranded structures known as G-quadruplexes. These structures have been proposed to play important biological roles and have been identified as potential drug targets. As a result, there is increasing interest in developing small molecules that can bind to G-quadruplexes. So far, these efforts have been mostly limited to conventional batch synthesis. Furthermore, no quick on-line method to assess new G-quadruplex binders has been developed. Herein, we report on two new microfluidic platforms to: (a) readily prepare G-quadruplex binders (based on metal complexes) in flow, quantitatively and without the need for purification before testing; (b) a microfluidic platform (based on FRET melting assays of DNA) that enables the real-time and on-line assessment of G-quadruplex binders in continuous flow...
April 14, 2018: Chemical Science
https://www.readbyqxmd.com/read/29780258/influence-of-different-ex-vivo-cell-culture-methods-on-the-proliferation-and-anti-tumor-activity-of-cytokine-induced-killer-cells-from-gastric-cancer-patients
#10
Bin Shi, Aixia Sun, Xiaorui Zhang
Purpose: In cytokine-induced killer (CIK) cell therapy, the phenotypes and the numbers of CIK cells have a great influence on the therapeutic effects. This study aimed to investigate the effects of different ex vivo cell culture methods on the proliferation and cytotoxicity of CIK cells that were obtained from gastric cancer patients. Patients and methods: CIK precursor (Pre-CIK) cells were collected by either hydroxyethyl starch (HES) sedimentation (HES method, unpurified group) or Ficoll-Hypaque density gradient centrifugation (Ficoll method, purified group)...
2018: OncoTargets and Therapy
https://www.readbyqxmd.com/read/29780184/pore-scale-hydrodynamics-in-a-progressively-bioclogged-three-dimensional-porous-medium-3-d-particle-tracking-experiments-and-stochastic-transport-modeling
#11
M Carrel, V L Morales, M Dentz, N Derlon, E Morgenroth, M Holzner
Biofilms are ubiquitous bacterial communities that grow in various porous media including soils, trickling, and sand filters. In these environments, they play a central role in services ranging from degradation of pollutants to water purification. Biofilms dynamically change the pore structure of the medium through selective clogging of pores, a process known as bioclogging. This affects how solutes are transported and spread through the porous matrix, but the temporal changes to transport behavior during bioclogging are not well understood...
March 2018: Water Resources Research
https://www.readbyqxmd.com/read/29779667/terminal-alkyne-biosynthesis-in-marine-microbes
#12
Xuejun Zhu, Wenjun Zhang
The terminal alkyne is a readily derivatized functionality valued for its diverse applications in material synthesis, pharmaceutical science, and chemical biology. The synthetic biology routes to terminal alkynes are highly desired and yet underexplored. Some marine natural products contain a terminal alkyne functionality, and the discovery of the biosynthetic gene clusters for jamaicamide B and carmabin A marked the beginning of a new era in the understanding and engineering of terminal alkyne biosynthesis...
2018: Methods in Enzymology
https://www.readbyqxmd.com/read/29779664/pks-nrps-enzymology-and-structural-biology-considerations-in-protein-production
#13
Meredith A Skiba, Finn P Maloney, Qingyun Dan, Amy E Fraley, Courtney C Aldrich, Janet L Smith, W Clay Brown
The structural diversity and complexity of marine natural products have made them a rich and productive source of new bioactive molecules for drug development. The identification of these new compounds has led to extensive study of the protein constituents of the biosynthetic pathways from the producing microbes. Essential processes in the dissection of biosynthesis have been the elucidation of catalytic functions and the determination of 3D structures for enzymes of the polyketide synthases and nonribosomal peptide synthetases that carry out individual reactions...
2018: Methods in Enzymology
https://www.readbyqxmd.com/read/29779661/characterization-and-biochemical-assays-of-streptomyces-vanadium-dependent-chloroperoxidases
#14
Shaun M K McKinnie, Zachary D Miles, Bradley S Moore
Vanadium-dependent haloperoxidases (VHPOs) are fascinating enzymes that facilitate electrophilic halogen incorporation into electron-rich substrates, simply requiring vanadate, a halide source, and cosubstrate hydrogen peroxide for activity. Initially characterized in fungi and red algae, VHPOs were long believed to have limited regio-, chemo-, and enantioselectivity in the production of halogenated metabolites. However, the recent discovery of homologues in the biosynthetic gene clusters of the stereoselectively halogenated meroterpenoids from marine-derived Streptomyces bacteria has revised this paradigm...
2018: Methods in Enzymology
https://www.readbyqxmd.com/read/29779489/rap1-downregulation-by-mir-320c-reduces-platelet-activation-in-ex-vivo-storage
#15
Chintamani Atreya, Neetu Dahiya
BACKGROUND: A small GTPase protein, the Ras-related protein 1 (RAP1), abundant in platelets is known to be activated following agonist induced platelet activation, suggesting that RAP1 downregulation could in turn reduce platelet activation in storage. Our objective of this study is to identify RAP1 regulating miRNAs and their role in platelet activation during storage. METHODS: We applied MS2-TRAP (tagged RNA affinity purification) methodology to enrich miRNAs that target the 3' untranslated region (3'UTR) of RAP1 mRNA in two mammalian cell lines followed by miRNA identification by microarray of total RNA samples enriched for miRNAs...
May 20, 2018: MicroRNA
https://www.readbyqxmd.com/read/29779487/expression-purification-and-characterization-of-a-gii-4-norovirus-protease
#16
Benjamin D Kuiper, Bradley J Keusch, Joshua Holcomb, Kendall M Muzzarelli, Franck Amblard, Peng Liu, Shaoman Zhou, Iulia A Kovari, Zhe Yang, Raymond F Schinazi, Ladislau C Kovari
BACKGROUND: Noroviruses are the leading cause of acute gastroenteritis worldwide. Norovirus proteases, which are responsible for cleavage of the viral polyprotein, have become an attractive drug target to treat norovirus infections. Genogroup II (GII) noroviruses are responsible for a majority of outbreaks; however, limited data exist regarding GII norovirus proteases. METHOD: The Minerva virus protease (MVpro), a genogroup II genotype 4 (GII.4) norovirus protease was expressed in E...
May 20, 2018: Infectious Disorders Drug Targets
https://www.readbyqxmd.com/read/29779478/exosomes-in-therapy-engineering-pharmacokinetic-and-future-applications
#17
Claudia Arenaccio, Chiara Chiozzini, Flavia Ferrantelli, Patrizia Leone, Eleonora Olivetta, Maurizio Federico
BACKGROUND: Eukaryotic cells release vesicles of different sizes under both physiological and pathological conditions. On the basis of the respective biogenesis, extracellular vesicles are classified as apoptotic bodies, microvesicles, and exosomes. Among these, exosomes are considered tools for innovative therapeutic interventions, especially when engineered with effector molecules. The delivery functions of exosomes are favored by a number of typical features. These include their small size (i...
May 20, 2018: Current Drug Targets
https://www.readbyqxmd.com/read/29778875/purification-of-recombinant-trichodysplasia-spinulosa-associated-polyomavirus-vp1-derived-virus-like-particles-using-chromatographic-techniques
#18
Mindaugas Zaveckas, Karolis Goda, Danguole Ziogiene, Alma Gedvilaite
Trichodysplasia spinulosa-associated polyomavirus (TSPyV) has been linked to a rare and recently characterized skin disease occurring in immunocompromised patients. In analogy with other polyomaviruses, the major capsid protein VP1 of TSPyV can self-assemble into virus-like particles (VLPs). VLPs are increasingly applied for the vaccination and diagnostics. Mostly, non-scalable and labor intensive ultracentrifugation-based techniques are used for the purification of VLPs. In this work, we developed a purification procedure for TSPyV VP1 VLPs based on two chromatographic steps, ion-exchange monolith and core bead chromatography...
May 9, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29778873/bioanalysis-of-ibrutinib-and-its-dihydrodiol-and-glutathione-cycle-metabolites-by-liquid-chromatography-tandem-mass-spectrometry
#19
J J M Rood, P J A Dormans, M J van Haren, J H M Schellens, J H Beijnen, R W Sparidans
Ibrutinib is a targeted covalent inhibitor frequently used for the treatment of various lymphomas. In addition to oxidative metabolism, it is metabolized through glutathione coupling. The quantitative insight into this kind of metabolism is scarce, and tools for quantitation are lacking. The non-oxidative metabolism could prove a more prominent role when oxidative metabolism is impaired. Also, in-vitro studies could over-estimate the effect of CYP450-inhibition. To gain quantitative insight into this relatively unknown biotransformation pathway of the drug we have developed a validated simple, fast and sensitive bio-analytical assay for ibrutinib, dihydrodiol-ibrutinib, and the glutathione, cysteinylglycine and cysteine conjugates of ibrutinib in human plasma...
May 11, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/29778683/biochar-enhances-plant-growth-and-nutrient-removal-in-horizontal-subsurface-flow-constructed-wetlands
#20
Kuno Kasak, Jaak Truu, Ivika Ostonen, Jürgen Sarjas, Kristjan Oopkaup, Päärn Paiste, Margit Kõiv-Vainik, Ülo Mander, Marika Truu
Biochar has shown great potential as an amendment to improve soil quality and promote plant growth, as well as to adsorb pollutants from water. However, information about the effect of biochar on the wastewater treatment efficiency in horizontal subsurface flow (HSSF) constructed wetlands (CWs) is still scarce. In this study, we assessed the effect of biochar amendment on the purification efficiency of pretreated municipal wastewater in planted (Typha latifolia) experimental horizontal subsurface flow filters filled with lightweight expanded clay aggregates (LECA)...
May 17, 2018: Science of the Total Environment
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