Mass spectrometry (MS)-based proteomics has great potential for overcoming the limitations of antibody-based immunoassays for antibody-independent, comprehensive, and quantitative proteomic analysis of single cells. Indeed, recent advances in nanoscale sample preparation have enabled effective processing of single cells. In particular, the concept of using boosting/carrier channels in isobaric labeling to increase the sensitivity in MS detection has also been increasingly used for quantitative proteomic analysis of small-sized samples including single cells. However, the full potential of such boosting/carrier approaches has not been significantly explored, nor has the resulting quantitation quality been carefully evaluated. Herein, we have further evaluated and optimized our recent boosting to amplify signal with isobaric labeling (BASIL) approach, originally developed for quantifying phosphorylation in small number of cells, for highly effective analysis of proteins in single cells. This improved BASIL (iBASIL) approach enables reliable quantitative single-cell proteomics analysis with greater proteome coverage by carefully controlling the boosting-to-sample ratio (e.g. in general <100x) and optimizing MS automatic gain control (AGC) and ion injection time settings in MS/MS analysis (e.g. 5E5 and 300 ms, respectively, which is significantly higher than that used in typical bulk analysis). By coupling with a nanodroplet-based single cell preparation (nanoPOTS) platform, iBASIL enabled identification of ~2500 proteins and precise quantification of ~1500 proteins in the analysis of 104 FACS-isolated single cells, with the resulting protein profiles robustly clustering the cells from three different acute myeloid leukemia cell lines. This study highlights the importance of carefully evaluating and optimizing the boosting ratios and MS data acquisition conditions for achieving robust, comprehensive proteomic analysis of single cells.

6 November 2019 , Volume 95, Number 6

The gang muck in to discuss the hottest tech topics of the week (as is tradition). Deputy Editor of Macworld UK David Price asks Jim Martin what was hot at MWC, where phones are birthed or something. Then Techworld Online Editor Thomas MacAuley chats about the government's digital strategy. First question: does it have one? Finally Tamlin Magee, Computerworld UK's Online Editor explains how to hack a robot and why people will almost certainly use this for EVIL. Thanks for listening.  

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In quantitative proteomics work, the differences in expression of many separate proteins are routinely examined to test for significant differences between treatments. This leads to the multiple hypothesis testing problem: when many separate tests are performed many will be significant by chance and be false positive results. Statistical methods such as the false discovery rate (FDR) method that deal with this problem have been disseminated for more than one decade. However a survey of proteomics journals shows that such tests are not widely implemented in one commonly used technique, quantitative proteomics using two-dimensional electrophoresis (2-DE). We outline a selection of multiple hypothesis testing methods, including some that are well known and some lesser known, and present a simple strategy for their use by the experimental scientist in quantitative proteomics work generally. The strategy focuses on the desirability of simultaneous use of several different methods, the choice and emphasis dependent on research priorities and the results in hand. This approach is demonstrated using case scenarios with experimental and simulated model data.

Invitation Only Research Event

Event participants

Oscar Jonsson, Director, Stockholm Free World Forum (Frivärld)
Mathieu Boulègue, Research Fellow, Russia and Eurasia Programme, Chatham House
Chair: ​Alice Billon-Galland, Research Associate, Europe Programme, Chatham House

14 February 2020

Madison Bumgarner has made it known he's not a fan of the "opener," a strategy of utilizing a pitcher -- usually a reliever -- to get the first few outs of a game before bringing in a pitcher who would usually start or pitch long relief.

Invitation Only Research Event

Event participants

Dr Kori Schake, Resident Scholar and Director of Foreign and Defense Policy Studies, American Enterprise Institute 
Chair: Dr Leslie Vinjamuri, Director, US and Americas Programme

Invitation Only Research Event

Event participants

Oscar Jonsson, Director, Stockholm Free World Forum (Frivärld)
Mathieu Boulègue, Research Fellow, Russia and Eurasia Programme, Chatham House
Chair: ​Alice Billon-Galland, Research Associate, Europe Programme, Chatham House

Research Event

Event participants

Lord Tu'ivakano, Prime Minister of the Kingdom of Tonga

Since the Rays debuted "the opener" last May, no fewer than eight teams have helped transform the concept from a fad into a mini-movement. Consider the Orioles a candidate to dive in next.

The UK government published its report Childhood Obesity: a Plan for Action, in August 2016. A new analysis article takes them to task for the inadequacy of that response to a growing problem. Neena Modi is a professor of neonatal medicine, at Imperial College London, and president of the Royal College of Paediatrics and Child Health, and joins...

Friction made strategy+business's 2019 list of Best Business Books, one of just three in the management category.

The post Best Business Books of 2019 – Strategy+Business appeared first on Neuromarketing.

A century ago, Edward Noble sold billions of Life Savers in a few years with a different approach to marketing mints.

The post How One Simple Strategy Changed the Candy Industry appeared first on Neuromarketing.

'"Safe and well" outlines the actions the Government of South Australia is taking to support families at risk of entering the child protection system, protect children from harm when they come into care, and invest in young people to leave care with opportunities for a bright future.

A $59 million investment in software developer Social Solutions aims to ease the flow of data among schools and social service providers.

Speech by Mr Agustín Carstens, General Manager of the BIS, at the Fourteenth ASBA-BCBS-FSI High-level Meeting on Global and Regional Supervisory Priorities, Lima, 1 October 2019.

Once the search has begun, something will be found

We can adapt flexibly to environment changes and search for the most appropriate rule to a context. The orbital prefrontal cortex (PFo) has been associated with decision making, rule generation and maintenance, and more generally has been considered important for behavioral flexibility. To better understand the neural mechanisms underlying the flexible behavior, we studied the ability to generate a switching signal in monkey PFo when a strategy is changed. In the strategy task, we used a visual cue to instruct two male rhesus monkeys either to repeat their most recent choice (i.e., stay strategy) or to change it (i.e., shift strategy). To identify the strategy switching-related signal, we compared nonswitch and switch trials, which cued the same or a different strategy from the previous trial, respectively. We found that the switching-related signal emerged during the cue presentation and it was combined with the strategy signal in a subpopulation of cells. Moreover, the error analysis showed that the activity of the switch-related cells reflected whether the monkeys erroneously switched or not the strategy, rather than what was required for that trial. The function of the switching signal could be to prompt the use of different strategies when older strategies are no longer appropriate, conferring the ability to adapt flexibly to environmental changes. In our task, the switching signal might contribute to the implementation of the strategy cued, overcoming potential interference effects from the strategy previously cued. Our results support the idea that ascribes to PFo an important role for behavioral flexibility.

SIGNIFICANCE STATEMENT We can flexibly adapt our behavior to a changing environment. One of the prefrontal areas traditionally associated with the ability to adapt to new contingencies is the orbital prefrontal cortex (PFo). We analyzed the switching related activity using a strategy task in which two rhesus monkeys were instructed by a visual cue either to repeat or change their most recent choice, respectively using a stay or a shift strategy. We found that PFo neurons were modulated by the strategy switching signal, pointing to the importance of PFo in behavioral flexibility by generating control over the switching of strategies.

"It is unfortunate that it has come to pass that we have to collectively and overtly publicly declare that we are unified," says Alberta Medical Association president Christine Molnar.

A PUBLIC health professor has warned that the Scottish Government’s strategy to escape the lockdown will encounter “a big problem” unless test results can be provided quicker.

THE SCOTTISH Government is reviewing its Covid-19 testing strategy after the Deputy First Minster has been left “frustrated” by reports home care workers have been told to travel to the other side of Scotland for tests.

A $59 million investment in software developer Social Solutions aims to ease the flow of data among schools and social service providers.

NEIL Mackay (“Johnson? Sturgeon? When it comes to coronavirus they are both the same”, The Herald, May 5) lambasts Nicola Sturgeon and Boris Johnson for both taking an almost identical approach in their fight against Covid-19, somehow implying that this is in itself a fault.

William Leou, a 6th grade science teacher at the Brooklyn School for Collaborative Studies, uses an organizational worksheet to help students draw arguments from evidence.

Chris Knutson, an 8th grade history teacher at Horning Middle School in Waukesha, Wis., shares how he incorporates learning conferences into his lessons.

Preterm infants can be successfully resuscitated with <100% oxygen (O₂); however, initiation with room air remains controversial. Current Neonatal Resuscitation Program (NRP) guidelines suggest using air or blended O₂ to titrate O₂ to meet target preductal saturation goals.

This is the first trial to compare a limited O₂ strategy to target NRP–recommended transitional goal saturations versus a high O₂ strategy in preterm infants. The limited O₂ strategy decreased integrated excess oxygen and oxidative stress and improved respiratory outcomes. (Read the full article)

A South African working abroad, able to save R36 000 per month, hopeful that he will continue these earnings, seeks the best investment strategy. Investment expert Elian Wiener responds.

Customer relationship management spent the last couple of years going social. Make sure your CRM strategy incorporates this important new capability in 2020.

There's no one-stop shop to get everything you need from education policy, politics, and practice, writes academic Deven E. Carlson.

Advanced analytics programs can incorporate geospatial data. Learn how such data can be used to augment local marketing plans

Commission must safeguard standards in the wake of Covid-19

In March 2014, the government approved two projects for setting up semiconductor units and also created an empowered committee (EC) for implementation of the projects.

Efforts are underway to develop a vaccine for Coronavirus on an urgent basis, but it’s a long-drawn process

MicroStrategy Intelligence Server and Web version 10.4 suffers from remote code execution, cross site scripting, server-side request forgery, and information disclosure vulnerabilities.

A more detailed look at fDi's judges’ top five American Cities of the Future 2019/20 for FDI strategy. Naomi Davies reports.