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Improving Identification of In-organello Protein-Protein Interactions Using an Affinity-enrichable, Isotopically Coded, and Mass Spectrometry-cleavable Chemical Crosslinker

Karl A. T. Makepeace
Apr 1, 2020; 19:624-639
Research




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XG Firewall is now available on AWS

We are extremely pleased to announce the availability of XG Firewall on Amazon Web Services (AWS) public cloud infrastructure.




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Improving Identification of In-organello Protein-Protein Interactions Using an Affinity-enrichable, Isotopically Coded, and Mass Spectrometry-cleavable Chemical Crosslinker [Research]

An experimental and computational approach for identification of protein-protein interactions by ex vivo chemical crosslinking and mass spectrometry (CLMS) has been developed that takes advantage of the specific characteristics of cyanurbiotindipropionylsuccinimide (CBDPS), an affinity-tagged isotopically coded mass spectrometry (MS)-cleavable crosslinking reagent. Utilizing this reagent in combination with a crosslinker-specific data-dependent acquisition strategy based on MS2 scans, and a software pipeline designed for integrating crosslinker-specific mass spectral information led to demonstrated improvements in the application of the CLMS technique, in terms of the detection, acquisition, and identification of crosslinker-modified peptides. This approach was evaluated on intact yeast mitochondria, and the results showed that hundreds of unique protein-protein interactions could be identified on an organelle proteome-wide scale. Both known and previously unknown protein-protein interactions were identified. These interactions were assessed based on their known sub-compartmental localizations. Additionally, the identified crosslinking distance constraints are in good agreement with existing structural models of protein complexes involved in the mitochondrial electron transport chain.




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Glucose availability but not changes in pancreatic hormones sensitizes hepatic AMPK activity during nutritional transition in rodents [Metabolism]

The cellular energy sensor AMP-activated protein kinase (AMPK) is a metabolic regulator that mediates adaptation to nutritional variations to maintain a proper energy balance in cells. We show here that suckling-weaning and fasting-refeeding transitions in rodents are associated with changes in AMPK activation and the cellular energy state in the liver. These nutritional transitions were characterized by a metabolic switch from lipid to glucose utilization, orchestrated by modifications in glucose levels and the glucagon/insulin ratio in the bloodstream. We therefore investigated the respective roles of glucose and pancreatic hormones on AMPK activation in mouse primary hepatocytes. We found that glucose starvation transiently activates AMPK, whereas changes in glucagon and insulin levels had no impact on AMPK. Challenge of hepatocytes with metformin-induced metabolic stress strengthened both AMPK activation and cellular energy depletion under limited-glucose conditions, whereas neither glucagon nor insulin altered AMPK activation. Although both insulin and glucagon induced AMPKα phosphorylation at its Ser485/491 residue, they did not affect its activity. Finally, the decrease in cellular ATP levels in response to an energy stress was additionally exacerbated under fasting conditions and by AMPK deficiency in hepatocytes, revealing metabolic inflexibility and emphasizing the importance of AMPK for maintaining hepatic energy charge. Our results suggest that nutritional changes (i.e. glucose availability), rather than the related hormonal changes (i.e. the glucagon/insulin ratio), sensitize AMPK activation to the energetic stress induced by the dietary transition during fasting. This effect is critical for preserving the cellular energy state in the liver.




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Medicare details available on dark web is just tip of data breach iceberg

The next wave of government reform will have to focus on data management.




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Webinar: European Democracy in the Last 100 Years: Economic Crises and Political Upheaval

Members Event Webinar

6 May 2020 - 1:00pm to 2:00pm

Event participants

Pepijn Bergsen, Research Fellow, Europe Programme, Chatham House

Dr Sheri Berman, Professor of Political Science, Barnard College

Chair: Hans Kundnani, Senior Research Fellow, Europe Programme, Chatham House

 

In the last 100 years, global economic crises from the Great Depression of the 1930s to the 2008 financial crash have contributed to significant political changes in Europe, often leading to a rise in popularity for extremist parties and politics. As Europe contends with a perceived crisis of democracy - now compounded by the varied responses to the coronavirus outbreak - how should we understand the relationship between externally-driven economic crises, political upheaval and democracy?

The panellists will consider the parallels between the political responses to some of the greatest economic crises Europe has experienced in the last century. Given that economic crises often transcend borders, why does political disruption vary between democracies? What can history tell us about the potential political impact of the unfolding COVID-19-related economic crisis? And will the unprecedented financial interventions by governments across Europe fundamentally change the expectations citizens have of the role government should play in their lives?

This event is based on a recent article in The World Today by Hans Kundnani and Pepijn Bergsen who are both researchers in Chatham House's Europe Programme. 'Crawling from the Wreckage' is the first in a series of articles that look at key themes in European political discourse from the last century. You can read the article here

This event is open to Chatham House Members. Not a member? Find out more.




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Coronavirus Vaccine: Available For All, or When it's Your Turn?

4 May 2020

Professor David Salisbury CB

Associate Fellow, Global Health Programme
Despite high-level commitments and pledges to cooperate to ensure equitable global access to a coronavirus vaccine, prospects for fair distribution are uncertain.

2020-05-04-Vaccine-COVID-Brazil

Researcher in Brazil working on virus replication in order to develop a vaccine against the coronavirus. Photo by DOUGLAS MAGNO/AFP via Getty Images.

When the H1N1 influenza pandemic struck in 2009, some industrialized countries were well prepared. Many countries’ preparedness plans had focused on preparing for an influenza pandemic and based on earlier alerts over the H5N1 ‘bird flu’ virus, countries had made advanced purchase or ‘sleeping’ contracts for vaccine supplies that could be activated as soon as a pandemic was declared. Countries without contracts scrambled to get supplies after those that already had contracts received their vaccine.

Following the 2009 pandemic, the European Union (EU) developed plans for joint-purchase vaccine contracts that any member state could join, guaranteeing the same price per dose for everyone. In 2009, low-income countries were unable to get the vaccine until manufacturers agreed to let 10 per cent of their production go to the World Health Organization (WHO).

The situation for COVID-19 could be even worse. No country had a sleeping contract in place for a COVID-19 vaccine since nobody had anticipated that the next pandemic would be a coronavirus, not an influenza virus. With around 80 candidate vaccines reported to be in development, choosing the right one will be like playing roulette.

These candidates will be whittled down as some will fail at an early stage of development and others will not get to scale-up for manufacturing. All of the world’s major vaccine pharmaceutical companies have said that they will divert resources to manufacture COVID-19 vaccines and, as long as they choose the right candidate for production, they have the expertise and the capacity to produce in huge quantities.

From roulette to a horse race

Our game now changes from roulette to a horse race, as the probability of winning is a matter of odds not a random chance. Countries are now able to try to make contracts alone or in purchasing consortia with other states, and with one of the major companies or with multiple companies. This would be like betting on one of the favourites.

For example, it has been reported that Oxford University has made an agreement with pharmaceutical company AstraZeneca, with a possibility of 100 million doses being available by the end of 2020. If the vaccine works and those doses materialize, and are all available for the UK, then the UK population requirements will be met in full, and the challenge becomes vaccinating everyone as quickly as possible.

Even if half of the doses were reserved for the UK, all those in high-risk or occupational groups could be vaccinated rapidly. However, as each major manufacturer accepts more contracts, the quantity that each country will get diminishes and the time to vaccinate the at-risk population gets longer.

At this point, it is not known how manufacturers will respond to requests for vaccine and how they will apportion supplies between different markets. You could bet on an outsider. You study the field and select a biotech that has potential with a good production development programme and a tie-in with a smaller-scale production facility.

If other countries do not try to get contracts, you will get your vaccine as fast as manufacturing can be scaled up; but because it is a small manufacturer, your supplies may take a long time. And outsiders do not often win races. You can of course, depending on your resources, cover several runners and try to make multiple contracts. However, you take on the risk that some will fail, and you may have compromised your eventual supply.

On April 24, the WHO co-hosted a meeting with the president of France, the president of the European Commission and the Bill & Melinda Gates Foundation. It brought together heads of state and industry leaders who committed to ‘work towards equitable global access based on an unprecedented level of partnership’. They agreed ‘to create a strong unified voice, to build on past experience and to be accountable to the world, to communities and to one another’ for vaccines, testing materials and treatments.

They did not, however, say how this will be achieved and the absence of the United States was notable. The EU and its partners are hosting an international pledging conference on May 4 that aims to raise €7.5 billion in initial funding to kick-start global cooperation on vaccines. Co-hosts will be France, Germany, Italy, the United Kingdom, Norway and Saudi Arabia and the priorities will be ‘Test, Treat and Prevent’, with the latter dedicated to vaccines.

Despite these expressions of altruism, every government will face the tension between wanting to protect their own populations as quickly as possible and knowing that this will disadvantage poorer countries, where health services are even less able to cope. It will not be a vote winner to offer a share in available vaccine to less-privileged countries.

The factories for the biggest vaccine manufacturers are in Europe, the US and India. Will European manufacturers be obliged by the EU to restrict sales first to European countries? Will the US invoke its Defense Production Act and block vaccine exports until there are stocks enough for every American? And will vaccine only be available in India for those who can afford it?

The lessons on vaccine availability from the 2009 influenza pandemic are clear: vaccine was not shared on anything like an equitable basis. It remains to be seen if we will do any better in 2020.




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Problem Notes for SAS®9 - 65903: You see a "java.lang.IllegalArgumentException" error in the log file when you use the IFRS9_Cycle workflow template in SAS Solution for IFRS 9

The problem occurs on a content release on the SAS Risk Governance Framework.




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Problem Notes for SAS®9 - 65872: You see a "java.lang.IllegalArgumentException" error in the log file when you use the CECL_Cycle workflow template in SAS Solution for CECL

The problem occurs on a content release on the SAS Risk Governance Framework.




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ADAM10 and ADAM17 proteases mediate proinflammatory cytokine-induced and constitutive cleavage of endomucin from the endothelial surface [Membrane Biology]

Contact between inflammatory cells and endothelial cells (ECs) is a crucial step in vascular inflammation. Recently, we demonstrated that the cell-surface level of endomucin (EMCN), a heavily O-glycosylated single-transmembrane sialomucin, interferes with the interactions between inflammatory cells and ECs. We have also shown that, in response to an inflammatory stimulus, EMCN is cleared from the cell surface by an unknown mechanism. In this study, using adenovirus-mediated overexpression of a tagged EMCN in human umbilical vein ECs, we found that treatment with tumor necrosis factor α (TNF-α) or the strong oxidant pervanadate leads to loss of cell-surface EMCN and increases the levels of the C-terminal fragment of EMCN 3- to 4-fold. Furthermore, treatment with the broad-spectrum matrix metalloproteinase inhibitor batimastat (BB94) or inhibition of ADAM metallopeptidase domain 10 (ADAM10) and ADAM17 with two small-molecule inhibitors, GW280264X and GI254023X, or with siRNA significantly reduced basal and TNFα-induced cell-surface EMCN cleavage. Release of the C-terminal fragment of EMCN by TNF-α treatment was blocked by chemical inhibition of ADAM10 alone or in combination with ADAM17. These results indicate that cell-surface EMCN undergoes constitutive cleavage and that TNF-α treatment dramatically increases this cleavage, which is mediated predominantly by ADAM10 and ADAM17. As endothelial cell-surface EMCN attenuates leukocyte–EC interactions during inflammation, we propose that EMCN is a potential therapeutic target to manage vascular inflammation.




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Get ready to watch: MLB.TV available for 2019

Spring Training is imminent, Opening Day is within sight and the big league season isn't complete for fans without a subscription to MLB.TV. The most comprehensive streaming service in professional sports is now available for the 2019 season.




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Talk Evidence - aggravating acronyms, a time to prescribe, and screening (again)

Talk Evidence is back, with your monthly take on the world of EBM with Duncan Jarvies and GPs Carl Heneghan (also director for the Centre of Evidence Based Medicine at the University of Oxford) and Helen Macdonald (also The BMJ's UK research Editor). This month Helen talks about the messy business of colon cancer screening - which modality is...




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Get ready to watch: MLB.TV available for 2019

Spring Training is imminent, Opening Day is within sight and the big league season isn't complete for fans without a subscription to MLB.TV. The most comprehensive streaming service in professional sports is now available for the 2019 season.




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Get ready to watch: MLB.TV available for 2019

Spring Training is imminent, Opening Day is within sight and the big league season isn't complete for fans without a subscription to MLB.TV. The most comprehensive streaming service in professional sports is now available for the 2019 season.




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Webinar: European Democracy in the Last 100 Years: Economic Crises and Political Upheaval

Members Event Webinar

6 May 2020 - 1:00pm to 2:00pm

Event participants

Pepijn Bergsen, Research Fellow, Europe Programme, Chatham House

Dr Sheri Berman, Professor of Political Science, Barnard College

Chair: Hans Kundnani, Senior Research Fellow, Europe Programme, Chatham House

 

In the last 100 years, global economic crises from the Great Depression of the 1930s to the 2008 financial crash have contributed to significant political changes in Europe, often leading to a rise in popularity for extremist parties and politics. As Europe contends with a perceived crisis of democracy - now compounded by the varied responses to the coronavirus outbreak - how should we understand the relationship between externally-driven economic crises, political upheaval and democracy?

The panellists will consider the parallels between the political responses to some of the greatest economic crises Europe has experienced in the last century. Given that economic crises often transcend borders, why does political disruption vary between democracies? What can history tell us about the potential political impact of the unfolding COVID-19-related economic crisis? And will the unprecedented financial interventions by governments across Europe fundamentally change the expectations citizens have of the role government should play in their lives?

This event is based on a recent article in The World Today by Hans Kundnani and Pepijn Bergsen who are both researchers in Chatham House's Europe Programme. 'Crawling from the Wreckage' is the first in a series of articles that look at key themes in European political discourse from the last century. You can read the article here

This event is open to Chatham House Members. Not a member? Find out more.




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Envie-nos a sua avaliação de nossos serviços

Você utilizou os nossos serviços e já tem algo a dizer sobre nós? Por favor, não hesite em nos enviar os seus comentários para que possamos continuar a aprimorar o nosso atendimento.




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Navajo Nation Reels Under Weight of Coronavirus and History of Neglect

Source:

The Navajo Nation now has the highest per-capita coronavirus infection rate after New York and New Jersey, but it has a fraction of the resources to treat and prevent the pandemic. "This has got to end," said president Jonathan Nez after federal relief funds arrived six weeks after they were promised and a week after the U.S. government missed a congressional deadline for distribution.






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Cleveland Cavaliers planning to reopen training facility Friday

The Cleveland Cavaliers are planning to reopen their training facility for limited individual workouts amid the coronavirus pandemic.




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New ADA standard, technical report available for comment by Feb. 17

The American Dental Association Standards Committee on Dental Informatics has approved two documents for circulation and comment.




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ADA standard on treating biofilm in waterlines available for comment

The American Dental Association Standards Committee on Dental Products has approved the document for circulation and comment.




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New HPI Industry Report now available

Dental spending reached $136 billion in 2018 — 3.7% of all health spending — and what the ADA Health Policy Institute is calling a “historic high” in the new HPI Industry Report released Feb. 4.




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ADA technical report on age assessment by dental analysis available for review

The ADA Standards Committee on Dental Informatics has approved the technical report for circulation and comment.




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Infection control webinar from ADA, OSAP available late March 20

On March 20, the ADA will release an on-demand webinar offering answers to frequently asked questions about infection control protocol and procedures related to COVID-19.




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ADA will share Paycheck Protection Program information as soon as it becomes available

The Association is waiting for clear guidance from the Small Business Administration on the best way to help dentists considering applying for Paycheck Protection Program 7(a) loans.




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Webinar on Small Business Administration loans available online

The ADA webinar, Small Business Administration Loans: Understanding the Options for Dentist Owners, is available online.




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SBA announces funding no longer available for Economic Injury Disaster Loans, Paycheck Protection Program

The Small Business Administration said that as of April 16, the agency is unable to accept any new applications for the Paycheck Protection Program or the Economic Injury Disaster Loans due to a lack of funding.




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Commercially Available Insulin Products Demonstrate Stability Throughout the Cold Supply Chain Across the U.S.

OBJECTIVE

A recent publication questioned the integrity of insulin purchased from U.S. retail pharmacies. We sought to independently validate the method used, isotope dilution solid-phase extraction (SPE) liquid chromatography mass spectrometry (LC-MS), and expand analysis to two U.S. Pharmacopeia (USP) methods (high-performance LC with ultraviolet detection and LC-MS).

RESEARCH DESIGN AND METHODS

Each method was used to evaluate nine insulin formulations, purchased at four pharmacies, within five geographic locations in the U.S.

RESULTS

All human and analog insulins measured by the USP methods (n = 174) contained the expected quantity of active insulin (100 ± 5 units/mL). When using isotope dilution SPE-LC-MS, units-per-milliliter values were well below product labeling due to unequal recovery of the internal standard compared with target insulin.

CONCLUSIONS

Insulin purchased from U.S. pharmacies is consistent with product labeling.




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Muscavado pavlova with chocolate and hazelnut mousse and Christmas cherries

I love Christmas my all time favourite time of year. It's also a great time for awesome summer sweet produce. Muscovado brown sugar is a treacle like flavour and makes a welcome texture to a traditional pavlova.




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Goat's Curd Bavarois with Cardamom Apples

Exotic, different, really cool flavours, pared-back dessert and it's not too sweet. Enjoy!




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CASSAVA LEAVES AND UGALI

Cassava leaves and Ugali - a traditional Congolese dish ... and much, much more. Gigi Mapatano




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2019 SJDM Conference program available online

SOCIETY FOR JUDGMENT AND DECISION MAKING CONFERENCE PROGRAM: DOWNLOAD AWAY The program for the 2019 SJDM conference (taking place this weekend (Nov 15-19, 2019) in Montreal, Canada) can be found in two places The SJDM home page (program link) The SJDM Newsletter (link) Please get it while it is hot.

The post 2019 SJDM Conference program available online appeared first on Decision Science News.




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2003 California-USC thriller available to all fans Saturday at 1:30 PT/ 2:30 MT on Pac-12 Now

Download the Pac-12 Now app to watch an epic, triple-overtime 2003 battle between USC and California this Saturday at 1:30 p.m. PT/ 2:30 p.m. MT. The game will be available to all fans.




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New edition of the Law Handbook available online

The Law handbook: your practical guide to the law in NSW is the single most important plain English guide to the law in




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Latest Law Handbook edition now available

Over 270 copies of The Law Handbook: Your Practical Guide to the Law in NSW — the leading plain English guide to the law




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To make terms of compromise a rule of Court or not? That is the question : an analysis of the options available to settle estate matters / presended by Christina Flourentzou, Supreme Court of South Australia..




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Newly available: Rare Books Collection




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[Brah atthukathapetavatthu]

19th century




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[Wellcome MS. Javanese 1]

1831




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[Wellcome MS. Javanese 2]




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[Wellcome MS. Javanese 3]




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[Wellcome MS. Javanese 4]




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[Wellcome MS. Javanese 5]




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[Wellcome MS. Javanese 6]

1820




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[Wellcome MS. Javanese 7]




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[Wellcome MS. Javanese 8]




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[Wellcome MS. Javanese 9]




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[Wellcome MS. Javanese 10]




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[Wellcome MS. Javanese 11]




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Des ruptures utérines pendant le travail de l'accouchement, considérées surtout au point de vue des symptômes et du traitement / par le Dr Jacques Jolly.

Paris : Delahaye, 1873.




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Des tumeurs synoviales de la partie infërieure de l'avant-bras, de la face palmaire du poignet, et de la main ... / par L.-M. Michon.

Londres : Paris, 1851.