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https://www.readbyqxmd.com/read/29423800/using-fluidigm-c1-to-generate-single-cell-full-length-cdna-libraries-for-mrna-sequencing
#1
Robert Durruthy-Durruthy, Manisha Ray
Single-cell RNA sequencing has evolved into a benchmark application to study cellular heterogeneity, advancing our understanding of cellular differentiation, disease progression, and gene regulation in a multitude of research areas. The generation of high-quality cDNA, an important step in the experimental workflow when generating sequence-ready libraries, is critical to maximizing data quality. Here we describe a strategy that uses a microfluidic device (i.e., the C1™ IFC) to synthesize full-length cDNA from single cells in a fully automated, nanoliter-scale format...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29397309/potentials-and-pitfalls-of-inverse-fluorescence-correlation-spectroscopy
#2
REVIEW
Stefan Wennmalm
Inverse Fluorescence Correlation Spectroscopy (iFCS) is a variant of FCS where unlabeled particles in solution, or domains in membranes, displace their surrounding, signal-generating molecules and thereby generate fluctuations. iFCS has to date been applied to unlabeled as well as labeled particles and protein molecules, using fluorescence as well as Raman scattering as a signal source, in diffraction-limited detection volumes as well as in nano-wells, and on fixed surfaces as well as in lipid bilayers. This review describes these applications and discusses the potentials and pitfalls when using iFCS...
January 31, 2018: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29395345/diagnostic-potential-of-imaging-flow-cytometry
#3
Minh Doan, Ivan Vorobjev, Paul Rees, Andrew Filby, Olaf Wolkenhauer, Anne E Goldfeld, Judy Lieberman, Natasha Barteneva, Anne E Carpenter, Holger Hennig
Imaging flow cytometry (IFC) captures multichannel images of hundreds of thousands of single cells within minutes. IFC is seeing a paradigm shift from low- to high-information-content analysis, driven partly by deep learning algorithms. We predict a wealth of applications with potential translation into clinical practice.
January 30, 2018: Trends in Biotechnology
https://www.readbyqxmd.com/read/29392698/single-cell-rna-sequencing-of-glioblastoma-cells
#4
Rajeev Sen, Igor Dolgalev, N Sumru Bayin, Adriana Heguy, Aris Tsirigos, Dimitris G Placantonakis
Single-cell RNA sequencing (sc-RNASeq) is a recently developed technique used to evaluate the transcriptome of individual cells. As opposed to conventional RNASeq in which entire populations are sequenced in bulk, sc-RNASeq can be beneficial when trying to better understand gene expression patterns in markedly heterogeneous populations of cells or when trying to identify transcriptional signatures of rare cells that may be underrepresented when using conventional bulk RNASeq. In this method, we describe the generation and analysis of cDNA libraries from single patient-derived glioblastoma cells using the C1 Fluidigm system...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29337047/imaging-flow-cytometry-a-method-for-examining-dynamic-native-foxo1-localization-in-human-lymphocytes
#5
Molly K Hritzo, Jean-Paul Courneya, Amit Golding
While flow cytometry can reliably assess surface and intracellular marker expression within small cell populations, it does not provide any information on protein localization. Several key transcription factors (TF) downstream of lymphocyte surface receptors are regulated by nuclear versus cytoplasmic localization, and one such TF is Forkhead box O1 (FOXO1). FOXO1 integrates antigen-binding, co-receptor activation and metabolic signals in lymphocytes, leading to proliferation and differentiation. Importantly, the nuclear or cytoplasmic localization of FOXO1 is key for gene expression leading to different lymphocyte phenotypes...
January 11, 2018: Journal of Immunological Methods
https://www.readbyqxmd.com/read/29330078/intraoperative-flow-cytometry-enables-the-differentiation-of-primary-central-nervous-system-lymphoma-from-glioblastoma
#6
Shunichi Koriyama, Masayuki Nitta, Takahiro Shioyama, Takashi Komori, Takashi Maruyama, Takakazu Kawamata, Yoshihiro Muragaki
OBJECTIVE: Accurate preoperative and intraoperative differentiation between primary central nervous system lymphoma (PCNSL) and glioblastoma (GBM) is sometimes difficult. Distinguishing between these tumors during surgery is important because surgical treatment is different between the two tumors. In this study, we established a new method of intraoperative differentiation between PCNSL and GBM using intraoperative flow cytometry (iFC), and we retrospectively tested whether iFC was useful for the intraoperative diagnosis of PCNSL and GBM...
January 9, 2018: World Neurosurgery
https://www.readbyqxmd.com/read/29305851/quantitative-assessment-of-inflow-malposition-in-two-continuous-flow-left-ventricular-assist-devices
#7
Erik N Sorensen, Zachary N Kon, Erika D Feller, Si M Pham, Bartley P Griffith
BACKGROUND: We previously investigated preoperative variables associated with qualitative inflow cannula (IFC) malposition in the Heartmate II continuous-flow left ventricular assist device. In this report, we assess IFC malposition quantitatively in recipients of both the Heartmate II and the Heartware and examine its association with device thrombosis. METHODS: Malposition was quantified based on angular deviation from a hypothetical ideal IFC position in two orthogonal computed tomography (CT) imaging planes...
January 3, 2018: Annals of Thoracic Surgery
https://www.readbyqxmd.com/read/29282846/meta-analysis-of-transcutaneous-electrical-nerve-stimulation-for-relief-of-spinal-pain
#8
REVIEW
L Resende, E Merriwether, É P Rampazo, D Dailey, J Embree, J Deberg, R E Liebano, K A Sluka
We conducted a systematic review and meta-analysis analysing the existing data on transcutaneous electrical nerve stimulation (TENS) or interferential current (IFC) for chronic low back pain (CLBP) and/or neck pain (CNP) taking into account intensity and timing of stimulation, examining pain, function and disability. Seven electronic databases were searched for TENS or IFC treatment in non-specific CLBP or CNP. Four reviewers independently selected randomized controlled trials (RCTs) of TENS or IFC intervention in adult individuals with non-specific CLBP or CNP...
December 27, 2017: European Journal of Pain: EJP
https://www.readbyqxmd.com/read/29278632/influence-of-the-type-of-plateletpheresis-on-the-value-of-corpuscular-elements-in-the-blood-donors
#9
Elvedin Landžo, Josip Petrović, Maja Karin, Ivan Tomić, Danijel Pravdić
INTRODUCTION: During the plateletpheresis procedure the number of trombocites in the donor's blood significantly decreases, and the levels of the other components of blood as hematocrit, hemoglobin, and leukocyte diminish as well. Influence of the type of procedure DN-CFCS and SN-ICFS it is one of the factors that affects the decrease of the levels of HCT, Hgb and WBC. In this study, our goal was to see the difference in the value of HCT, Hgb, WBC, and platelets after the plateletfphresis process between DN-CFCS and SN-IFCS on the same cell separator - Fenval AMICUS...
December 2017: Psychiatria Danubina
https://www.readbyqxmd.com/read/29235506/differential-contributions-of-the-middle-frontal-gyrus-functional-connectivity-to-literacy-and-numeracy
#10
Maki S Koyama, David O'Connor, Zarrar Shehzad, Michael P Milham
Literacy and numeracy equally affect an individual's success in and beyond schools, but these two competencies tend to be separately examined, particularly in neuroimaging studies. The current resting-state fMRI study examined the neural correlates of literacy and numeracy in the same sample of healthy adults. We first used an exploratory "Multivariate Distance Matrix Regression" (MDMR) approach to examine intrinsic functional connectivity (iFC), highlighting the middle frontal gyrus (MFG) for both competencies...
December 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29232069/medicare-program-cy-2018-updates-to-the-quality-payment-program-and-quality-payment-program-extreme-and-uncontrollable-circumstance-policy-for-the-transition-year-final-rule-with-comment-period-and-interim-final-rule-with-comment-period
#11
(no author information available yet)
The Medicare Access and CHIP Reauthorization Act of 2015 (MACRA) established the Quality Payment Program for eligible clinicians. Under the Quality Payment Program, eligible clinicians can participate via one of two tracks: Advanced Alternative Payment Models (APMs); or the Merit-based Incentive Payment System (MIPS). We began implementing the Quality Payment Program through rulemaking for calendar year (CY) 2017. This final rule with comment period provides updates for the second and future years of the Quality Payment Program...
November 16, 2017: Federal Register
https://www.readbyqxmd.com/read/29230329/regionally-specific-regulation-of-sensorimotor-network-connectivity-following-tactile-improvement
#12
Stefanie Heba, Melanie Lenz, Tobias Kalisch, Oliver Höffken, Lauren M Schweizer, Benjamin Glaubitz, Nicolaas A J Puts, Martin Tegenthoff, Hubert R Dinse, Tobias Schmidt-Wilcke
Correlations between inherent, task-free low-frequency fluctuations in the blood oxygenation level-dependent (BOLD) signals of the brain provide a potent tool to delineate its functional architecture in terms of intrinsic functional connectivity (iFC). Still, it remains unclear how iFC is modulated during learning. We employed whole-brain resting-state magnetic resonance imaging prior to and after training-independent repetitive sensory stimulation (rSS), which is known to induce somatosensory cortical reorganization...
2017: Neural Plasticity
https://www.readbyqxmd.com/read/29225336/psychiatric-polygenic-risk-associates-with-cortical-morphology-and-functional-organization-in-aging
#13
Annie Lee, Mojun Shen, Anqi Qiu
Common brain abnormalities in cortical morphology and functional organization are observed in psychiatric disorders and aging, reflecting shared genetic influences. This preliminary study aimed to examine the contribution of a polygenetic risk for psychiatric disorders (PRScross) to aging brain and to identify molecular mechanisms through the use of multimodal brain images, genotypes, and transcriptome data. We showed age-related cortical thinning in bilateral inferior frontal cortex (IFC) and superior temporal gyrus and alterations in the functional connectivity between bilateral IFC and between right IFC and right inferior parietal lobe as a function of PRScross...
December 11, 2017: Translational Psychiatry
https://www.readbyqxmd.com/read/29208929/the-choice-of-cryopreservation-method-affects-immune-compatibility-of-human-cardiovascular-matrices
#14
Maria Schneider, Christof Stamm, Kelvin G M Brockbank, Ulrich A Stock, Martina Seifert
Conventional frozen cryopreservation (CFC) is currently the gold standard for cardiovascular allograft preservation. However, inflammation and structural deterioration limit transplant durability. Ice-free cryopreservation (IFC) already demonstrated matrix structure preservation combined with attenuated immune responses. In this study, we aim to explore the mechanisms of this diminished immunogenicity in vitro. First, we characterized factors released by human aortic tissue after CFC and IFC. Secondly, we analyzed co-cultures with human peripheral blood mononuclear cells, purified monocytes, T cells and monocyte-derived macrophages to examine functional immune effects triggered by the tissue or released cues...
December 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29198814/a-high-content-imaging-flow-cytometry-approach-to-study-mitochondria-in-t-cells-mitotracker-green-fm-dye-concentration-optimization
#15
Namrata Gautam, Shvetha Sankaran, John A Yason, Kevin S W Tan, Nicholas R J Gascoigne
Mitochondria, the powerhouse of the cell, are known to remodel their membrane structures through the process of fusion or fission. Studies have indicated that T cells adopt different energy metabolic phenotypes, namely oxidative phosphorylation and glycolysis depending on whether they are naïve, effector and memory T cells. It has recently been shown that changes in mitochondrial morphology dictate T cell fate via regulation of their metabolism. Our keen interest in T cell function and metabolism led us to explore and establish a method to study mitochondria in live T cells through a novel high content approach called Imaging Flow Cytometry (IFC)...
November 30, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29186380/impact-of-t-cell-mediated-immune-response-on-xenogeneic-heart-valve-transplantation-short-term-success-and-mid-term-failure
#16
Anna C Biermann, Julia Marzi, Eva Brauchle, Maria Schneider, Angela Kornberger, Sherif Abdelaziz, Julian L Wichmann, Christophe T Arendt, Eike Nagel, Kelvin G M Brockbank, Martina Seifert, Katja Schenke-Layland, Ulrich A Stock
OBJECTIVES: Allogeneic frozen cryopreserved heart valves (allografts or homografts) are commonly used in clinical practice. A major obstacle for their application is the limited availability in particular for paediatrics. Allogeneic large animal studies revealed that alternative ice-free cryopreservation (IFC) results in better matrix preservation and reduced immunogenicity. The objective of this study was to evaluate xenogeneic (porcine) compared with allogeneic (ovine) IFC heart valves in a large animal study...
November 27, 2017: European Journal of Cardio-thoracic Surgery
https://www.readbyqxmd.com/read/29177509/repetitive-behaviors-in-autism-are-linked-to-imbalance-of-corticostriatal-connectivity-a-functional-connectivity-mri-study
#17
Angela E Abbott, Annika Linke, Aarti Nair, Afrooz Jahedi, Laura A Alba, Christopher L Keown, Inna Fishman, Ralph-Axel Müller
The neural underpinnings of repetitive behaviors (RBs) in autism spectrum disorders (ASDs), ranging from cognitive to motor characteristics, remain unknown. We assessed RB symptomatology in 50 ASD and 52 typically developing (TD) children and adolescents (ages 8-17 years), examining intrinsic functional connectivity (iFC) of corticostriatal circuitry, which is important for reward-based learning and integration of emotional, cognitive, and motor processing, and considered impaired in ASDs. Connectivity analyses were performed for three functionally distinct striatal seeds (limbic, frontoparietal, motor)...
November 21, 2017: Social Cognitive and Affective Neuroscience
https://www.readbyqxmd.com/read/29153950/highly-sensitive-electrochemical-nuclear-factor-kappa-b-aptasensor-based-on-target-induced-dual-signal-ratiometric-and-polymerase-assisted-protein-recycling-amplification-strategy
#18
Kanfu Peng, Pan Xie, Zhe-Han Yang, Ruo Yuan, Keqin Zhang
In this work, an amplified electrochemical ratiometric aptasensor for nuclear factor kappa B (NF-κB) assay based on target binding-triggered ratiometric signal readout and polymerase-assisted protein recycling amplification strategy is described. To demonstrate the effect of "signal-off" and "signal-on" change for the dual-signal electrochemical ratiometric readout, the thiol-hairpin DNA (SH-HD) hybridizes with methylene blue (MB)-modified protection DNA (MB-PD) to form capture probes, which is rationally introduced for the construction of the assay platform...
October 26, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29145074/ratiometric-electrochemical-assay-for-sensitive-detecting-microrna-based-on-dual-amplification-mechanism-of-duplex-specific-nuclease-and-hybridization-chain-reaction
#19
Yan-Hong Yuan, Bao-Zhu Chi, Shao-Hua Wen, Ru-Ping Liang, Zhi-Mei Li, Jian-Ding Qiu
We propose a ratiometric electrochemical assay for detecting microRNA (miRNA) on the basis of dual-amplification mechanism by using distinguishable electrochemical signals from thionine (Thi) and ferrocene (Fc). The thiol-modified and ferrocene-labeled hairpin capture probes (CP) are first immobilized on an Au electrode via Au-S reaction. The target miRNA hybridizes with CP and unfolding the hairpin structure of CP to form miRNA-DNA duplexes. Then, kamchatka crab duplex specific nuclease (DSN) specifically cleaves the DNA in miRNA-DNA duplexes, leading to the release of miRNA and another cleaves cycle, meanwhile, numerous Fc leaves away from the electrode surface and leads to the signal-off of Fc...
November 8, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29136539/an-examination-of-lower-limb-asymmetry-in-ankle-isometric-force-control
#20
Sheng-Che Yen, Laura C Olsavsky, Caitlin M Cloonan, Amelia R Llanos, Kara J Dwyer, Mohsen Nabian, Amir Bahador Farjadian
While asymmetries have been observed between the dominant and non-dominant legs, it is unclear whether they have different abilities in isometric force control (IFC). The purpose of this study was to compare ankle IFC between the legs. IFC is important for stabilization rather than object manipulation, and people typically use their non-dominant leg for stabilization tasks. Additionally, studies suggested that a limb can better acquire a motor task when the control mechanism of the task is related to what the limb is specialized for...
November 11, 2017: Human Movement Science
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