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https://www.readbyqxmd.com/read/28549322/high-throughput-continuous-production-of-liposomes-using-hydrodynamic-flow-focusing-microfluidic-devices
#1
Mariano Michelon, Davi Rocha Bernardes Oliveira, Guilherme de Figueiredo Furtado, Lucimara Gaziola de la Torre, Rosiane Lopes Cunha
The microfluidic hydrodynamic flow-focusing is a simple technique for nanoscale liposome formation that provides several advantages compared to the traditional manufacturing techniques. This work aimed to perform a systematic study of the liposome formation using planar microfluidic devices with different channel aspect-ratios, as an alternative to enhance the throughput of liposome synthesis. In general, liposomes with a low polydispersity and a precise control of the size were successfully produced from alteration of the flow rate ratio and channel aspect-ratio...
May 11, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28549288/ecological-effects-of-soil-properties-and-metal-concentrations-on-the-composition-and-diversity-of-microbial-communities-associated-with-land-use-patterns-in-an-electronic-waste-recycling-region
#2
Wencheng Wu, Changxun Dong, Jiahui Wu, Xiaowen Liu, Yingxin Wu, Xianbin Chen, Shixiao Yu
Soil microbes play vital roles in ecosystem functions, and soil microbial communities may be strongly structured by land use patterns associated with electronic waste (e-waste) recycling activities, which can increase the heavy metal concentration in soils. In this study, a suite of soils from five land use types (paddy field, vegetable field, dry field, forest field, and e-waste recycling site) were collected in Longtang Town, Guangdong Province, South China. Soil physicochemical properties and heavy metal concentrations were measured, and the indigenous microbial assemblages were profiled using 16S rRNA high-throughput sequencing and clone library analyses...
May 23, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28549267/the-construction-of-a-novel-nucleic-acids-detection-microplatform-based-on-the-nset-for-one-step-detecting-tk1-dna-and-microrna-21
#3
Jian Zhang, Qian Zhao, Yudong Wu, Bo Zhang, Weipan Peng, Jiafang Piao, Yurui Zhou, Weichen Gao, Xiaoqun Gong, Jin Chang
Microbeads-based microchip technology has become the potential for a new generation of nucleic acids detection in a high-throughput and sensitive manner. However the specificity and operational complexity limit the microchip applied in nucleic acids detection. Herein, in this work, we designed a kind of gold-nanoparticles coated polystyrene microbeads as microplatform conjugating with the molecular beacons as probes. Due to the nanoparticle surface energy transfer of gold-nanoparticles, the fluorescence of dye on one end of molecular beacons was effectively quenched...
May 23, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28549193/label-free-direct-localization-and-relative-quantitation-of-the-rna-nucleobase-methylations-m6a-m5c-m3u-and-m5u-by-top-down-mass-spectrometry
#4
Heidelinde Glasner, Christian Riml, Ronald Micura, Kathrin Breuker
Nucleobase methylations are ubiquitous posttranscriptional modifications of ribonucleic acids (RNA) that can substantially increase the structural diversity of RNA in a highly dynamic fashion with implications for gene expression and human disease. However, high throughput, deep sequencing does not generally provide information on posttranscriptional modifications (PTMs). A promising alternative approach for the characterization of PTMs, i.e. their identification, localization, and relative quantitation, is top-down mass spectrometry (MS)...
May 26, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28549128/atad3-gene-cluster-deletions-cause-cerebellar-dysfunction-associated-with-altered-mitochondrial-dna-and-cholesterol-metabolism
#5
Radha Desai, Ann E Frazier, Romina Durigon, Harshil Patel, Aleck W Jones, Ilaria Dalla Rosa, Nicole J Lake, Alison G Compton, Hayley S Mountford, Elena J Tucker, Alice L R Mitchell, Deborah Jackson, Abdul Sesay, Miriam Di Re, Lambert P van den Heuvel, Derek Burke, David Francis, Sebastian Lunke, George McGillivray, Simone Mandelstam, Fanny Mochel, Boris Keren, Claude Jardel, Anne M Turner, P Ian Andrews, Jan Smeitink, Johannes N Spelbrink, Simon J Heales, Masakazu Kohda, Akira Ohtake, Kei Murayama, Yasushi Okazaki, Anne Lombès, Ian J Holt, David R Thorburn, Antonella Spinazzola
Although mitochondrial disorders are clinically heterogeneous, they frequently involve the central nervous system and are among the most common neurogenetic disorders. Identifying the causal genes has benefited enormously from advances in high-throughput sequencing technologies; however, once the defect is known, researchers face the challenge of deciphering the underlying disease mechanism. Here we characterize large biallelic deletions in the region encoding the ATAD3C, ATAD3B and ATAD3A genes. Although high homology complicates genomic analysis of the ATAD3 defects, they can be identified by targeted analysis of standard single nucleotide polymorphism array and whole exome sequencing data...
June 1, 2017: Brain: a Journal of Neurology
https://www.readbyqxmd.com/read/28548903/microenvironment-of-a-tumor-organoid-system-enhances-hepatocellular-carcinoma-malignancy-related-hallmarks
#6
Yang Wang, Kazuki Takeishi, Zhao Li, Eduardo Cervantes-Alvarez, Alexandra Collin de l'Hortet, Jorge Guzman-Lepe, Xiao Cui, Jiye Zhu
Organ-like microenviroment and 3-dimensional (3D) cell culture conformations have been suggested as promising approaches to mimic in a micro-scale a whole organ cellular functions and interactions present in vivo. We have used this approach to examine biologic features of hepatocellular carcinoma (HCC) cells. In this study, we demonstrate that hepatocellular carcinoma (HCC) cells, fibroblasts, endothelial cells and extracellular matrix can generate organoid-like spheroids that enhanced numerous features of human HCC observed in vivo...
May 26, 2017: Organogenesis
https://www.readbyqxmd.com/read/28548572/analytical-validation-of-a-reference-laboratory-elisa-for-the-detection-of-feline-leukemia-virus-p27-antigen
#7
Jesse S Buch, Genevieve H Clark, Roberta Cahill, Brendon Thatcher, Peter Smith, Ramaswamy Chandrashekar, Christian M Leutenegger, Thomas P O'Connor, Melissa J Beall
Feline leukemia virus (FeLV) is an oncogenic retrovirus of cats. Immunoassays for the p27 core protein of FeLV aid in the detection of FeLV infections. Commercial microtiter-plate ELISAs have rapid protocols and visual result interpretation, limiting their usefulness in high-throughput situations. The purpose of our study was to validate the PetChek FeLV 15 ELISA, which is designed for the reference laboratory, and incorporates sequential, orthogonal screening and confirmatory protocols. A cutoff for the screening assay was established with 100% accuracy using 309 feline samples (244 negative, 65 positive) defined by the combined results of FeLV PCR and an independent reference p27 antigen ELISA...
May 1, 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28548482/simultaneous-separation-and-washing-of-non-magnetic-particles-in-an-inertial-ferrofluid-water-co-flow
#8
Qi Chen, Di Li, Jianhan Lin, Mao-Hua Wang, Xiangchun Xuan
Magnetic fluids (e.g., paramagnetic solutions and ferrofluids) have been increasingly used for label-free separation of non-magnetic particles in microfluidic devices. Their biocompatibility, however, becomes a concern in high-throughput or large-volume applications. One way to potentially resolve this issue is re-suspending the particles that are separated in a magnetic fluid immediately into a biocompatible buffer. We demonstrate herein the proof-of-principle of the first integration of negative magnetophoresis and inertial focusing for a simultaneous separation and washing of non-magnetic particles in co-flowing ferrofluid and water streams...
May 26, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28548238/a-quantitative-property-property-relationship-for-the-internal-diffusion-coefficients-of-organic-compounds-in-solid-materials
#9
Lei Huang, Peter Fantke, Alexi Ernstoff, Olivier Jolliet
Indoor releases of organic chemicals encapsulated in solid materials are major contributors to human exposures and are directly related to the internal diffusion coefficient in solid materials. Existing correlations to estimate the diffusion coefficient are only valid for a limited number of chemical-material combinations. This paper develops and evaluates a quantitative property-property relationship (QPPR) to predict diffusion coefficients for a wide range of organic chemicals and materials. We first compiled a training dataset of 1103 measured diffusion coefficients for 158 chemicals in 32 consolidated material types...
May 26, 2017: Indoor Air
https://www.readbyqxmd.com/read/28548214/the-performance-of-malbac-and-mda-methods-in-the-identification-of-concurrent-mutations-and-aneuploidy-screening-to-diagnose-beta-thalassaemia-disorders-at-the-single-and-multiple-cell-levels
#10
WeiQiang Liu, HuiMin Zhang, Dan Hu, SiJia Lu, XiaoFang Sun
AIM: To select an optimal whole-genome amplification (WGA) method to improve the efficiency of the preimplantation genetic diagnosis and screening (PGD/PGS) of beta-thalassaemia disorders. METHODS: Fifty-seven fibroblast samples with defined beta-thalassaemia variations and forty-eight single-blastomere samples were amplified from single-, two-, and five-cell samples by multiple annealing and looping-based amplification cycles (MALBAC) and the multiple displacement amplification (MDA) method...
May 26, 2017: Journal of Clinical Laboratory Analysis
https://www.readbyqxmd.com/read/28547825/dual-redundant-sequencing-strategy-full-length-gene-characterisation-of-1056-novel-and-confirmatory-hla-alleles
#11
V Albrecht, C Zweiniger, V Surendranath, K Lang, G Schöfl, A Dahl, S Winkler, V Lange, I Böhme, A H Schmidt
The high-throughput department of DKMS Life Science Lab encounters novel human leukocyte antigen (HLA) alleles on a daily basis. To characterise these alleles, we have developed a system to sequence the whole gene from 5'- to 3'-UTR for the HLA loci A, B, C, DQB1 and DPB1 for submission to the European Molecular Biology Laboratory - European Nucleotide Archive (EMBL-ENA) and the IPD-IMGT/HLA Database. Our workflow is based on a dual redundant sequencing strategy. Using shotgun sequencing on an Illumina MiSeq instrument and single molecule real-time (SMRT) sequencing on a PacBio RS II instrument, we are able to achieve highly accurate HLA full-length consensus sequences...
May 25, 2017: HLA
https://www.readbyqxmd.com/read/28547520/small-scale-intraspecific-patterns-of-adaptive-immunogenetic-polymorphisms-and-neutral-variation-in-lake-superior-lake-trout
#12
Shauna M Baillie, Riley R Hemstock, Andrew M Muir, Charles C Krueger, Paul Bentzen
Many fishes express high levels of intraspecific variability, often linked to resource partitioning. Several studies show that a species' evolutionary trajectory of adaptive divergence can undergo reversals caused by changes in its environment. Such a reversal in neutral genetic and morphological variation among lake trout Salvelinus namaycush ecomorphs appears to be underway in Lake Superior. However, a water depth gradient in neutral genetic divergence was found to be associated with intraspecific diversity in the lake...
May 25, 2017: Immunogenetics
https://www.readbyqxmd.com/read/28546810/a-downmodulated-microrna-profiling-in-patients-with-gastric-cancer
#13
Tao Zhang, Chang Liu, Shi Huang, Yuanping Ma, Jiansong Fang, Yuanneng Chen
Objective. Here, we aim to investigate the microRNA (miR) profiling in human gastric cancer (GC). Methods. Tumoral and matched peritumoral gastric specimens were collected from 12 GC patients who underwent routine surgery. A high-throughput miR sequencing method was applied to detect the aberrantly expressed miRs in a subset of 6 paired samples. The stem-loop quantitative real-time polymerase chain reaction (qRT-PCR) assay was subsequently performed to confirm the sequencing results in the remaining 6 paired samples...
2017: Gastroenterology Research and Practice
https://www.readbyqxmd.com/read/28546554/high-throughput-resistance-profiling-of-plasmodium-falciparum-infections-based-on-custom-dual-indexing-and-illumina-next-generation-sequencing-technology
#14
Sidsel Nag, Marlene D Dalgaard, Poul-Erik Kofoed, Johan Ursing, Marina Crespo, Lee O'Brien Andersen, Frank Møller Aarestrup, Ole Lund, Michael Alifrangis
Genetic polymorphisms in P. falciparum can be used to indicate the parasite's susceptibility to antimalarial drugs as well as its geographical origin. Both of these factors are key to monitoring development and spread of antimalarial drug resistance. In this study, we combine multiplex PCR, custom designed dual indexing and Miseq sequencing for high throughput SNP-profiling of 457 malaria infections from Guinea-Bissau, at the cost of 10 USD per sample. By amplifying and sequencing 15 genetic fragments, we cover 20 resistance-conferring SNPs occurring in pfcrt, pfmdr1, pfdhfr, pfdhps, as well as the entire length of pfK13, and the mitochondrial barcode for parasite origin...
May 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28546429/structural-studies-unravel-the-active-conformation-of-apo-ror%C3%AE-t-nuclear-receptor-and-a-common-inverse-agonism-of-two-diverse-classes-of-ror%C3%AE-t-inhibitors
#15
Xiang Li, Marie Anderson, Delphine Collin, Ingo Muegge, John Wan, Debra Brennan, Stanley Kugler, Donna Terenzio, Charles Kennedy, Siqi Lin, Mark E Labadia, Brian Cook, Robert Hughes, Neil A Farrow
The nuclear receptor retinoid acid-related orphan receptor γt (RORγt) is a master regulator of the Th17/IL-17 pathway that plays crucial roles in the pathogenesis of autoimmunity. RORγt has recently emerged as a highly promising target for treatment of a number of autoimmune diseases. Through high-throughput screening we previously identified several classes of inverse agonists for RORγt. Here, we report the crystal structures for the ligand binding domain of RORγt in both apo and ligand-bound states. We show that apo RORγt adopts an active conformation capable of recruiting coactivator peptides and present a detailed analysis of the structural determinants that stabilize helix 12 (H12) of RORγt in the active state in the absence of a ligand...
May 25, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28546383/drosophila-simulans-a-species-with-improved-resolution-in-evolve-and-resequence-studies
#16
Neda Barghi, Raymond Tobler, Viola Nolte, Christian Schlötterer
The combination of experimental evolution with high-throughput sequencing of pooled individuals - i.e. Evolve and Resequence; E&R - is a powerful approach to study adaptation from standing genetic variation under controlled, replicated conditions. Nevertheless, E&R studies in Drosophila melanogaster have frequently resulted in inordinate numbers of candidate SNPs, particularly for complex traits. Here, we contrast the genomic signature of adaptation following ~60 generations in a novel hot environment for D...
May 25, 2017: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/28545524/an-unsupervised-learning-approach-for-tracking-mice-in-an-enclosed-area
#17
Jakob Unger, Mike Mansour, Marcin Kopaczka, Nina Gronloh, Marc Spehr, Dorit Merhof
BACKGROUND: In neuroscience research, mouse models are valuable tools to understand the genetic mechanisms that advance evidence-based discovery. In this context, large-scale studies emphasize the need for automated high-throughput systems providing a reproducible behavioral assessment of mutant mice with only a minimum level of manual intervention. Basic element of such systems is a robust tracking algorithm. However, common tracking algorithms are either limited by too specific model assumptions or have to be trained in an elaborate preprocessing step, which drastically limits their applicability for behavioral analysis...
May 25, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28545467/expression-of-deleted-atoxic-atypical-recombinant-beta2-toxin-in-a-baculovirus-system-and-production-of-polyclonal-and-monoclonal-antibodies
#18
Anna Serroni, Chiara Francesca Magistrali, Giovanni Pezzotti, Luca Bano, Martina Pellegrini, Giulio Severi, Chiara Di Pancrazio, Mirella Luciani, Manuela Tittarelli, Silvia Tofani, Antonio De Giuseppe
BACKGROUND: Clostridium perfringens is an important animal and human pathogen that can produce more than 16 different major and minor toxins. The beta-2 minor toxin (CPB2), comprising atypical and consensus variants, appears to be involved in both human and animal enterotoxaemia syndrome. The exact role of CPB2 in pathogenesis is poorly investigated, and its mechanism of action at the molecular level is still unknown because of the lack of specific reagents such as monoclonal antibodies against the CPB2 protein and/or the availability of a highly purified antigen...
May 25, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28545462/sequence-based-prediction-of-protein-protein-interaction-using-a-deep-learning-algorithm
#19
Tanlin Sun, Bo Zhou, Luhua Lai, Jianfeng Pei
BACKGROUND: Protein-protein interactions (PPIs) are critical for many biological processes. It is therefore important to develop accurate high-throughput methods for identifying PPI to better understand protein function, disease occurrence, and therapy design. Though various computational methods for predicting PPI have been developed, their robustness for prediction with external datasets is unknown. Deep-learning algorithms have achieved successful results in diverse areas, but their effectiveness for PPI prediction has not been tested...
May 25, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28545393/quantgenius-implementation-of-a-decision-support-system-for-qpcr-based-gene-quantification
#20
Špela Baebler, Miha Svalina, Marko Petek, Katja Stare, Ana Rotter, Maruša Pompe-Novak, Kristina Gruden
BACKGROUND: Quantitative molecular biology remains a challenge for researchers due to inconsistent approaches for control of errors in the final results. Due to several factors that can influence the final result, quantitative analysis and interpretation of qPCR data are still not trivial. Together with the development of high-throughput qPCR platforms, there is a need for a tool allowing for robust, reliable and fast nucleic acid quantification. RESULTS: We have developed "quantGenius" ( http://quantgenius...
May 25, 2017: BMC Bioinformatics
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