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https://www.readbyqxmd.com/read/28918538/genetic-manipulation-of-borrelia-spp
#1
Dan Drecktrah, D Scott Samuels
The spirochetes Borrelia (Borreliella) burgdorferi and Borrelia hermsii, the etiologic agents of Lyme disease and relapsing fever, respectively, cycle in nature between an arthropod vector and a vertebrate host. They have extraordinarily unusual genomes that are highly segmented and predominantly linear. The genetic analyses of Lyme disease spirochetes have become increasingly more sophisticated, while the age of genetic investigation in the relapsing fever spirochetes is just dawning. Molecular tools available for B...
September 17, 2017: Current Topics in Microbiology and Immunology
https://www.readbyqxmd.com/read/28918473/complete-genome-sequences-of-two-insect-specific-flaviviruses
#2
Jermilia Charles, Chandra S Tangudu, Andrew E Firth, Bradley J Blitvich
We determined the complete genomic sequences of two previously discovered insect-specific flaviviruses, Marisma mosquito virus (MMV) and Nanay virus (NANV), using a combination of high-throughput sequencing, reverse transcription-polymerase chain reaction, 5' and 3' rapid amplification of cDNA ends and Sanger sequencing. Complete polyprotein amino acid sequence alignments revealed that the closest known relatives of MMV and NANV are Donggang virus (89% identity, 95% similarity) and Nounané virus (53% identity, 70% similarity), respectively...
September 16, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28918158/recombinant-expression-and-characterization-of-lucilia-cuprina-cyp6g3-activity-and-binding-properties-toward-multiple-pesticides
#3
Matthew J Traylor, Jong-Min Baek, Katelyn E Richards, Roberto Fusetto, W Huang, Peter Josh, Zhengzhong Chen, Padma Bollapragada, Richard A J O'Hair, Philip Batterham, Elizabeth M J Gillam
The Australian sheep blowfly, Lucilia cuprina, is a primary cause of sheep flystrike and a major agricultural pest. Cytochrome P450 enzymes have been implicated in the resistance of L. cuprina to several classes of insecticides. In particular, CYP6G3 is a L. cuprina homologue of Drosophila melanogaster CYP6G1, a P450 known to confer multi-pesticide resistance. To investigate the basis of resistance, a bicistronic Escherichia coli expression system was developed to co-express active L. cuprina CYP6G3 and house fly (Musca domestica) P450 reductase...
September 13, 2017: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/28918098/identifying-novel-factor-xiia-inhibitors-with-pca-ga-svm-developed-vhts-models
#4
Jonathan Jun Feng Chen, Donald P Visco
There currently is renewed interest in blood clotting Factor XII as a potential target for thrombosis inhibition. Historically untargeted, there is little drug information with which to start drug candidate searches. Typical high-throughput screening can identify potential drug candidates, but is inefficient. Virtual high-throughput screening can be used to raise efficiency by focusing experimental efforts on compounds predicted to be active and is applied here to identify new Factor XIIa inhibitors. We combine principal component analysis, genetic algorithm and support vector machine to create the models used in the virtual high-throughput screening...
September 1, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28918093/topological-structure-determination-of-rna-using-small-angle-x-ray-scattering
#5
Yuba R Bhandari, Linxin Fan, Xianyang Fang, George Zaki, Eric A Stahlberg, Wei Jiang, Charles D Schwieters, Jason R Stagno, Yun-Xing Wang
Knowledge of RNA three-dimensional topological structures provides important insight into the relationship between RNA structural components and function. It is often likely that near complete sets of biochemical and biophysical data containing structural restraints are not available but one still wants to obtain knowledge about approximate topological folding of RNA. In this regard, general methods for determining such topological structures with minimum readily available restraints are lacking. Naked RNAs are difficult to crystallize and NMR spectroscopy is generally limited to small RNA fragments...
September 13, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28918052/a-ctla-4-antagonizing-dna-aptamer-with-antitumor-effect
#6
Bo-Tsang Huang, Wei-Yun Lai, Yi-Chung Chang, Jen-Wei Wang, Shauh-Der Yeh, Emily Pei-Ying Lin, Pan-Chyr Yang
The successful translation of cytotoxic T lymphocyte antigen-4 (CTLA-4) blockade has revolutionized the concept of cancer immunotherapy. Although monoclonal antibody therapeutics remain the mainstream in clinical practice, aptamers are synthetic oligonucleotides that encompass antibody-mimicking functions. Here, we report a novel high-affinity CTLA-4-antagonizing DNA aptamer (dissociation constant, 11.84 nM), aptCTLA-4, which was identified by cell-based SELEX and high-throughput sequencing. aptCTLA-4 is relatively stable in serum, promotes lymphocyte proliferation, and inhibits tumor growth in cell and animal models...
September 15, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28917797/a-rapid-and-specific-colorimetric-method-for-free-tryptophan-quantification
#7
Yinan Wu, Tianmin Wang, Chong Zhang, Xin-Hui Xing
Tryptophan is one of the eight essential amino acids and plays an important role in many biological processes. For its interaction with human health, environment and relevant commercial interest in biotechnology-based production, rapid and specific quantification method for this molecule accessible to common laboratories is badly needed. We herein reported a simple colorimetric method for free tryptophan quantification with 96-well-plate-level throughput. Our protocol firstly converted tryptophan to indole enzymatically by purified tryptophanases and then used reactivity of indole with hydroxylamine to form pink product with absorption peak at 530nm, enabling the quantification of tryptophan with simple spectrometry in just two hours...
January 1, 2018: Talanta
https://www.readbyqxmd.com/read/28917760/rapid-and-sensitive-serum-glucose-determination-using-chemical-labeling-coupled-with-black-phosphorus-assisted-laser-desorption-ionization-time-of-flight-mass-spectrometry
#8
Qing Wang, Lei Yu, Chu-Bo Qi, Jun Ding, Xiao-Mei He, Ren-Qi Wang, Yu-Qi Feng
Monitoring the concentration of blood glucose in patients is a key component of good medical diagnoses. Therefore, developing an accurate, rapid and sensitive strategy for monitoring blood glucose is of vital importance. We proposed a strategy for serum glucose determination combining 2-(4-boronobenzyl) isoquinolin-2-ium bromide chemical labeling with black phosphorus assisted laser desorption ionization-time of flight mass spectrometry (CL-BP/ALDI-TOF MS). The entire analytical process consisted of 1min of protein precipitation and 3min of chemical labeling in a microwave oven prior to the BP/ALDI-TOF MS analysis...
January 1, 2018: Talanta
https://www.readbyqxmd.com/read/28917730/direct-and-comprehensive-analysis-of-dyes-based-on-integrated-molecular-and-structural-information-via-laser-desorption-laser-postionization-mass-spectrometry
#9
Rong Liu, Zhibin Yin, Yixin Leng, Wei Hang, Benli Huang
Laser desorption laser postionization time-of-flight mass spectrometry (LDPI-TOFMS) was employed for direct analysis and determination of typical basic dyes. It was also used for the analysis and comprehensive understanding of complex materials such as blue ballpoint pen inks. Simultaneous emergences of fragmental and molecular information largely simplify and facilitate unambiguous identification of dyes via variable energy of 266nm postionization laser. More specifically, by optimizing postionization laser energy with the same energy of desorption laser, the structurally significant results show definite differences in the fragmentation patterns, which offer opportunities for discrimination of isomeric species with identical molecular weight...
January 1, 2018: Talanta
https://www.readbyqxmd.com/read/28917199/enhanced-phosphorus-and-ciprofloxacin-removal-in-a-modified-baf-system-by-configuring-fe-c-micro-electrolysis-investigation-on-pollutants-removal-and-degradation-mechanisms
#10
Longlong Zhang, Qinyan Yue, Kunlun Yang, Pin Zhao, Baoyu Gao
A modified biological aerated filter (BAF) system configured Fe-C micro electrolysis was applied to enhance phosphorus and ciprofloxacin (CIP) removal. A novel sludge ceramic and sintering ferric-carbon ceramic (SFC) were separately packed into a lab-scale BAF and Fe-C micro electrolysis reactor. The BAF and Fe-C micro electrolysis coupled system was operated about 230days. The enhancement of phosphorus and ciprofloxacin removals by Fe-C micro electrolysis, the degradation mechanisms of CIP and the variations of microbial population were investigated...
September 8, 2017: Journal of Hazardous Materials
https://www.readbyqxmd.com/read/28916864/sparticle-an-algorithm-for-the-analysis-of-filamentous-microorganisms-in-submerged-cultures
#11
Joost Willemse, Ferhat Büke, Dino van Dissel, Sanne Grevink, Dennis Claessen, Gilles P van Wezel
Streptomycetes are filamentous bacteria that produce a plethora of bioactive natural products and industrial enzymes. Their mycelial lifestyle typically results in high heterogeneity in bioreactors, with morphologies ranging from fragments and open mycelial mats to dense pellets. There is a strong correlation between morphology and production in submerged cultures, with small and open mycelia favouring enzyme production, while most antibiotics are produced mainly in pellets. Here we describe SParticle, a Streptomyces Particle analysis method that combines whole slide imaging with automated image analysis to characterize the morphology of submerged grown Streptomyces cultures...
September 15, 2017: Antonie Van Leeuwenhoek
https://www.readbyqxmd.com/read/28916818/identification-of-4-phenylquinolin-2-1h-one-as-a-specific-allosteric-inhibitor-of-akt
#12
Bill X Huang, Kenny Newcomer, Karl Kevala, Elena Barnaeva, Wei Zheng, Xin Hu, Samarjit Patnaik, Noel Southall, Juan Marugan, Marc Ferrer, Hee-Yong Kim
Akt plays a major role in tumorigenesis and the development of specific Akt inhibitors as effective cancer therapeutics has been challenging. Here, we report the identification of a highly specific allosteric inhibitor of Akt through a FRET-based high-throughput screening, and characterization of its inhibitory mechanism. Out of 373,868 compounds screened, 4-phenylquinolin-2(1H)-one specifically decreased Akt phosphorylation at both T308 and S473, and inhibited Akt kinase activity (IC50 = 6 µM) and downstream signaling...
September 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28916763/maternal-diabetes-causes-developmental-delay-and-death-in-early-somite-mouse-embryos
#13
Jing Zhao, Theodorus B M Hakvoort, Jan M Ruijter, Aldo Jongejan, Jan Koster, Sigrid M A Swagemakers, Aleksandar Sokolovic, Wouter H Lamers
Maternal diabetes causes congenital malformations and delays embryonic growth in the offspring. We investigated effects of maternal diabetes on mouse embryos during gastrulation and early organogenesis (ED7.5-11.5). Female mice were made diabetic with streptozotocin, treated with controlled-release insulin implants, and mated. Maternal blood glucose concentrations increased up to embryonic day (ED) 8.5. Maternal hyperglycemia induced severe growth retardation (approx.1 day) in 53% of the embryos on ED8.5, death in most of these embryos on ED9...
September 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28916716/deregulation-of-microrna-expression-in-purified-t-and-b-lymphocytes-from-patients-with-primary-sj%C3%A3-gren-s-syndrome
#14
Shu-Fang Wang-Renault, Saida Boudaoud, Gaétane Nocturne, Elodie Roche, Nelly Sigrist, Christian Daviaud, Andreas Bugge Tinggaard, Victor Renault, Jean-François Deleuze, Xavier Mariette, Jörg Tost
OBJECTIVE: MicroRNAs (miRNAs) play an important role in the pathogenesis of autoimmune diseases such as primary Sjögren's syndrome (pSS). This study is the first to investigate miRNA expression patterns in purified T and B lymphocytes from patients with pSS using a high-throughput quantitative PCR (qPCR) approach. METHODS: Two independent cohorts of both patients with pSS and controls, one for discovery and one for replication, were included in this study. CD4+ T cells and CD19+ B cells were isolated from peripheral blood mononuclear cells by magnetic microbeads and expression of miRNAs was profiled using the Exiqon Human miRNome panel I analysing 372 miRNAs...
September 15, 2017: Annals of the Rheumatic Diseases
https://www.readbyqxmd.com/read/28916650/differential-expression-of-mirnas-in-the-respiratory-tree-of-the-sea-cucumber-apostichopus-japonicus-under-hypoxia-stress
#15
Da Huo, Lina Sun, Xiaoni Li, Xiaoshang Ru, Shilin Liu, Libin Zhang, Lili Xing, Hongsheng Yang
Sea cucumber, an important economic species, has encountered high mortality since 2013 in northern China because of seasonal environmental stress such as hypoxia, high temperature, and low salinity. MicroRNAs (miRNAs) are important in regulating gene expression in marine organisms in response to environmental change. In the present study, high-throughput sequencing was used to investigate alterations in miRNA expression in the sea cucumber under different levels of dissolved oxygen (DO). Nine small RNA libraries were constructed from the sea cucumber respiratory trees...
September 15, 2017: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/28916508/development-of-a-whole-slide-imaging-system-on-smartphones-and-evaluation-with-frozen-section-samples
#16
Hong Yu, Feng Gao, Liren Jiang, Shuoxin Ma
BACKGROUND: The aim was to develop scalable Whole Slide Imaging (sWSI), a WSI system based on mainstream smartphones coupled with regular optical microscopes. This ultra-low-cost solution should offer diagnostic-ready imaging quality on par with standalone scanners, supporting both oil and dry objective lenses of different magnifications, and reasonably high throughput. These performance metrics should be evaluated by expert pathologists and match those of high-end scanners. OBJECTIVE: The aim was to develop scalable Whole Slide Imaging (sWSI), a whole slide imaging system based on smartphones coupled with optical microscopes...
September 15, 2017: JMIR MHealth and UHealth
https://www.readbyqxmd.com/read/28916394/a-useful-strategy-based-on-chromatographic-data-combined-with-quality-by-design-approach-for-food-analysis-applications-the-case-study-of-furanic-derivatives-in-sugarcane-honey
#17
Pedro Silva, Catarina L Silva, Rosa Perestrelo, Fernando M Nunes, José S Câmara
Sugarcane honey (SCH) is one of the Madeira Island products par excellence and it is now popular worldwide. Its sui generis and peculiar sensory properties, explained by a variety of volatile compounds including furanic derivatives (FDs), arise mainly from manufacturing and storage conditions. A simple high-throughput approach based on semi-automatic microextraction by packed sorbent (MEPS) combined with ultra-high performance liquid chromatography (UHPLC) was developed and validated for identification and quantification of target FDs in sugarcane honey...
September 8, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28916303/high-throughput-flow-cytometry-for-drug-discovery-principles-applications-and-case-studies
#18
REVIEW
Mei Ding, Karin Kaspersson, David Murray, Catherine Bardelle
Flow cytometry is a technology providing multiparametric analysis of single cells or other suspension particles. High-throughput (HT) flow cytometry has become an attractive screening platform for drug discovery. In this review, we highlight the recent HT flow cytometry applications, and then focus on HT flow cytometry deployment at AstraZeneca (AZ). Practical considerations for successful HT flow cytometry assay development and screening are provided based on experience from four project case studies at AZ...
September 12, 2017: Drug Discovery Today
https://www.readbyqxmd.com/read/28916278/microbial-reduction-of-vanadium-v-in-groundwater-interactions-with-coexisting-common-electron-acceptors-and-analysis-of-microbial-community
#19
Hui Liu, Baogang Zhang, Heyang Yuan, Yutong Cheng, Song Wang, Zhen He
Vanadium (V) pollution in groundwater has posed serious risks to the environment and public health. Anaerobic microbial reduction can achieve efficient and cost-effective remediation of V(V) pollution, but its interactions with coexisting common electron acceptors such as NO3(-), Fe(3+), SO4(2-) and CO2 in groundwater remain unknown. In this study, the interactions between V(V) reduction and reduction of common electron acceptors were examined with revealing relevant microbial community and identifying dominant species...
September 12, 2017: Environmental Pollution
https://www.readbyqxmd.com/read/28916103/controlled-synchronized-actuation-of-microdroplets-by-gravity-in-a-superhydrophobic-3d-printed-device
#20
P E Oomen, J P S H Mulder, E Verpoorte, R D Oleschuk
Droplet manipulation over open surfaces allows one to perform assays with a large degree of control and high throughput, making them appealing for applications in drug screening or (bio)analysis. However, the design, manufacturing and operation of these systems comes with high technical requirements. In this study we employ a commercial, low-friction, superhydrophobic coating, Ultra-Ever Dry(®), on a 3D-printed microfluidic device. The device features individual droplet compartments, which allow the manipulation of discrete droplets (10-50 μL) actuated by gravity alone...
October 2, 2017: Analytica Chimica Acta
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