There is a need to develop novel approaches to improve the balance between efficacy and toxicity for transcription factor–targeted therapies. In this study, we exploit context-dependent differences in RNA polymerase II processivity as an approach to improve the activity and limit the toxicity of the EWS-FLI1–targeted small molecule, mithramycin, for Ewing sarcoma. The clinical activity of mithramycin for Ewing sarcoma is limited by off-target liver toxicity that restricts the serum concentration to levels insufficient to inhibit EWS-FLI1. In this study, we perform an siRNA screen of the druggable genome followed by a matrix drug screen to identify mithramycin potentiators and a synergistic "class" effect with cyclin-dependent kinase 9 (CDK9) inhibitors. These CDK9 inhibitors enhanced the mithramycin-mediated suppression of the EWS-FLI1 transcriptional program leading to a shift in the IC₅₀ and striking regressions of Ewing sarcoma xenografts. To determine whether these compounds may also be liver protective, we performed a qPCR screen of all known liver toxicity genes in HepG2 cells to identify mithramycin-driven transcriptional changes that contribute to the liver toxicity. Mithramycin induces expression of the BTG2 gene in HepG2 but not Ewing sarcoma cells, which leads to a liver-specific accumulation of reactive oxygen species (ROS). siRNA silencing of BTG2 rescues the induction of ROS and the cytotoxicity of mithramycin in these cells. Furthermore, CDK9 inhibition blocked the induction of BTG2 to limit cytotoxicity in HepG2, but not Ewing sarcoma cells. These studies provide the basis for a synergistic and less toxic EWS-FLI1–targeted combination therapy for Ewing sarcoma.

RNA viruses form a dynamic distribution of mutant swarms (termed "quasispecies") due to the accumulation of mutations in the viral genome. The genetic diversity of a viral population is affected by several factors, including a bottleneck effect. Human-to-human transmission exemplifies a bottleneck effect, in that only part of a viral population can reach the next susceptible hosts. In the present study, two lineages of the rhesus rotavirus (RRV) strain of rotavirus A were serially passaged five times at a multiplicity of infection (MOI) of 0.1 or 0.001, and three phenotypes (infectious titer, cell binding ability, and specific growth rate) were used to evaluate the impact of a bottleneck effect on the RRV population. The specific growth rate values of lineages passaged under the stronger bottleneck (MOI of 0.001) were higher after five passages. The nucleotide diversity also increased, which indicated that the mutant swarms of the lineages under the stronger bottleneck effect were expanded through the serial passages. The random distribution of synonymous and nonsynonymous substitutions on rotavirus genome segments indicated that almost all mutations were selectively neutral. Simple simulations revealed that the presence of minor mutants could influence the specific growth rate of a population in a mutant frequency-dependent manner. These results indicate a stronger bottleneck effect can create more sequence spaces for minor sequences.

IMPORTANCE In this study, we investigated a bottleneck effect on an RRV population that may drastically affect the viral population structure. RRV populations were serially passaged under two levels of a bottleneck effect, which exemplified human-to-human transmission. As a result, the genetic diversity and specific growth rate of RRV populations increased under the stronger bottleneck effect, which implied that a bottleneck created a new space in a population for minor mutants originally existing in a hidden layer, which includes minor mutations that cannot be distinguished from a sequencing error. The results of this study suggest that the genetic drift caused by a bottleneck in human-to-human transmission explains the random appearance of new genetic lineages causing viral outbreaks, which can be expected according to molecular epidemiology using next-generation sequencing in which the viral genetic diversity within a viral population is investigated.

Background:

Despite widespread recognition that adherence to clinical preventive guidelines improves patient outcomes, clinicians struggle to implement guideline changes in a timely manner. Multiple factors influence guideline adoption and effective implementation. However, few studies evaluate their collective and inter-related effects. This qualitative study provides a comprehensive picture of the interplay between multiple factors on uptake of new or changed preventive guidelines.

Ayelen M. Blanco, Juan I. Bertucci, Jose L. Soengas, and Suraj Unniappan

This research assessed the direct effects of insulin on nutrient-sensing mechanisms in the brain of rainbow trout (Oncorhynchus mykiss) using an in vitro approach. Cultured hypothalamus and hindbrain were exposed to 1 µmol l^–1 insulin for 3 h, and signals involved in appetite regulation and nutrient-sensing mechanisms were measured. Additionally, the involvement of the phosphatidylinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in the actions of insulin was studied by using the inhibitor wortmannin. Treatment with insulin alone did not elicit many changes in the appetite regulators and nutrient-sensing-related genes and enzymes tested in the hypothalamus and hindbrain. However, we found that, when insulin and nutrients were added together, insulin reversed most of the effects exerted by nutrients alone, suggesting that insulin changes responsiveness to nutrients at the central level. Effects reversed by insulin included expression levels of genes related to the sensing of both glucose (slc2a2, slc5a1, gck, pck1, pklr, g6pcb, gys1, tas1r3 and nr1h3 in the hindbrain, and slc2a2, pklr and pck1 in the hypothalamus) and fatty acid (cd36 in the hindbrain, and cd36 and acly in the hypothalamus). Nutrient-induced changes in the activity of Acly and Cpt-1 in the hindbrain and of Pepck, Acly, Fas and Hoad in the hypothalamus were also reversed by insulin. Most of the insulin effects disappeared in the presence of wortmannin, suggesting the PI3K/Akt pathway is a mediator of the effects of insulin reported here. This study adds new information to our knowledge of the mechanisms regulating nutrient sensing in fish.

Imants G. Priede, Rhoderick W. Burgass, Manolis Mandalakis, Apostolos Spyros, Petros Gikas, Finlay Burns, and Jim Drewery

Whereas upper ocean pelagic sharks are negatively buoyant and must swim continuously to generate lift from their fins, deep-sea sharks float or swim slowly buoyed up by large volumes of low-density oils in their livers. Investigation of the Pressure, Volume, Temperature (PVT) relationships for liver oils of 10 species of deep-sea Chondrichthyes shows that the density difference between oil and seawater, remains almost constant with pressure down to full ocean depth (11 km, 1100 bar); theoretically providing buoyancy far beyond the maximum depth of occurrence (3700 m) of sharks. However, , does change significantly with temperature and we show that the combined effects of pressure and temperature can decrease buoyancy of oil by up to 10% between the surface and 3500 m depth across interfaces between warm southern and cold polar waters in the Rockall Trough in the NE Atlantic. This increases drag more than 10 fold compared with neutral buoyancy during horizontal slow swimming (0.1 m s^–1) but the effect becomes negligible at high speeds. Chondrichthyes generally experience positive buoyancy change during ascent and negative buoyancy change during descent but contrary effects can occur at interfaces between waters of different densities. During normal vertical migrations buoyancy changes are small, increasing slow-speed drag by no more than 2–3 fold. Equations and tables of density, pressure and temperature are provided for squalene and liver oils of Chimaeriformes (Harriotta raleighana, Chimaera monstrosa, Chimaera monstrosa), Squaliformes (Centrophorus squamosus, Deania calcea, Centroscymnus coelolepis, Centroscyllium fabricii, Etmopterus spinax) and Carcharhiniformes (Apristurus laurussonii, Galeus murinus).

Objective

To investigate the immunologic impact of a single cycle of rituximab (RTX) in children and adolescents with immune-mediated disorders, we evaluated B cells and immunoglobulin levels of 20 patients with neuroimmunologic, nephrologic, dermatologic, and rheumatologic disorders treated under recommended guidelines.

The number of onsite clinical microbiology laboratories in hospitals is decreasing, likely related to the business model for laboratory consolidation and labor shortages, and this impacts a variety of clinical practices, including that of banking isolates for clinical or epidemiologic purposes. To determine the impact of these trends, infectious disease (ID) physicians were surveyed regarding their perceptions of offsite services. Clinical microbiology practices for retention of clinical isolates for future use were also determined. Surveys were sent to members of the Infectious Diseases Society of America’s (IDSA) Emerging Infections Network (EIN). The EIN is a sentinel network of ID physicians who care for adult and/or pediatric patients in North America and who are members of IDSA. The response rate was 763 (45%) of 1,680 potential respondents. Five hundred forty (81%) respondents reported interacting with the clinical microbiology laboratory. Eighty-six percent of respondents thought an onsite laboratory very important for timely diagnostic reporting and ongoing communication with the clinical microbiologist. Thirty-five percent practiced in institutions where the core microbiology laboratory has been moved offsite, and an additional 7% (n = 38) reported that movement of core laboratory functions offsite was being considered. The respondents reported that only 24% of laboratories banked all isolates, with the majority saving isolates for less than 30 days. Based on these results, the trend toward centralized core laboratories negatively impacts the practice of ID physicians, potentially delays effective implementation of prompt and targeted care for patients with serious infections, and similarly adversely impacts infection control epidemiologic investigations.

It is important to identify, assess, and address current barriers to implementation of post-approval changes that are intended to ensure continued (uninterrupted) operations and drive innovation and continual improvement in a maximally efficient, agile, and flexible pharmaceutical manufacturing sector. Leveraging the International Conference for Harmonisation Quality Guideline Q10 provides regulatory relief when it comes to addressing changes related to excipients, specifically excipient supplier's name and address changes, which will ensure a sustainable, reliable global supply and the availability of high quality product to patients through the entire commercial lifecycle of a product without extensive regulatory oversight.

Low-dose adjuvant epigenetic therapy (AET) reduced metastasis and promoted survival in mouse models.

Background:

Reduction in breast density may be a biomarker of endocrine therapy (ET) efficacy. Our objective was to assess the impact of race on ET-related changes in volumetric breast density (VBD).

Fast-growing Melbourne payments company PromisePay is ramping its growth plans in the global platform economy.

Coffee has been linked to changes on our DNA that affect how active certain genes are. The finding may help explain some of coffee's touted health benefits

Enemies of the U.S. Army are now deliberately hiding targets behind mountain ridges, under bridges, in rocky crevices and other locations intended to elude state-of-the art GPS-guided artillery round attacks -- complicating U.S. efforts to pinpoint and destroy targets

The first changes to the UK's coronavirus lockdown will be "small" and "very carefully monitored" when the Prime Minister reveals his "roadmap" on Sunday.

Boris Johnson has said the Government will proceed with "maximum caution" when it comes to easing the coronavirus lockdown, with his spokesman adding that any changes next week will be "very limited".

British people should not expect major new freedoms on Monday, with "limited changes" expected to the coronavirus lockdown, a minister has said.

Publishers of several of Canada’s major newspapers signed a joint letter to the federal government this month, taking aim at the advertising revenue earned by Google and Facebook.

Comedian explained on her podcast that she hadn't realised what the name sounded like until recently

Pokemon GO (Free) from Niantic has been improving with letting people play the game in one place or at home. …

Continue reading "PokeCoin System Changes Are Coming to ‘Pokemon GO’ as Niantic Begins Testing in Australia"

Barcelona have announced changes to the club's board in a swift restructuring after six directors submitted their resignations to president Josep Maria Bartomeu last week.

Proposed changes to the Champions League could 'end the Premier League top-four race' with qualification restricted, according to reports.

A short glossary of the changes we’ve made to the Guardian’s style guide, for use by our journalists and editors when writing about the environment

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In addition to providing updated guidelines on which images our editors should use to illustrate the climate emergency, we have updated our style guide to introduce terms that more accurately describe the environmental crises facing the world. Our editor-in-chief, Katharine Viner, said: “We want to ensure that we are being scientifically precise, while also communicating clearly with readers on this very important issue”. These are the guidelines provided to our journalists and editors to be used in the production of all environment coverage across the Guardian’s website and paper:

Related: The urgency of climate crisis needed robust new language to describe it | Paul Chadwick

With the NFL heading into its next round of broadcasting negotiations, it could not afford to release a schedule full of unsightly seams and untenable sacrifices.

While the 2020 NFL season faces some uncertainties because of the coronavirus, the league leaned on a familiar road map in constructing the season schedule.

Broad modifications to current standards for treating acute stroke patients during the COVID-19 pandemic may be needed to preserve health care resources, limit disease spread and ensure optimal care, according to a Loyola Medicine neurologist.

The briefing comes just hours after Scotland's First Minister has said the lockdown in Scotland should be extended.

Welsh ministers understood to have agreed only 'cautious' changes to lockdown restrictions here in Wales

Culture Secretary Oliver Dowden said the government was also prepared to row back from easing restrictions if ‘things…get out of control’.

State Premier Daniel Andrews says lockdown measures will remain in place until at least Monday.

Premier Daniel Andrews says his Government will explain changes to the state's coronavirus restrictions on Monday, after the National Cabinet agrees to a three-step process of lifting restrictions to create a "COVID-safe economy".

Lockdown changes in England from next week will be "very limited", Boris Johnson's spokesman has confirmed.

The public have been told not to expect widespread changes to the coronavirus lockdown in England when Boris Johnson addresses the nation this weekend.

A leading voice in Formula One warns there is a serious threat of losing teams if big changes are not made to the competition.

By: Daniel Logan and Justine Johnson The Coronavirus Aid, Relief, and Economic Security Act, or “CARES” Act (Pub. L. No. 116–136) makes numerous changes to the Federal Food, Drug, and Cosmetic Act (FDCA).  The CARES Act made substantial changes related to the regulation of over-the-counter (OTC) drugs (see KKB’s alert here dedicated to summarizing these

The post Changes Made to the Federal Food, Drug, and Cosmetic Act by the CARES Act appeared first on Kleinfeld Kaplan & Becker LLP.

The Department will require Ticketmaster Entertainment Inc. to license its ticketing software, divest ticketing assets and subject itself to anti-retaliation provisions in order to proceed with its proposed merger with Live Nation Inc.

Today, the Justice Department announced that, pursuant to its agreement with the Illinois State Board of Education, the state has finalized administrative rules that will ensure school districts throughout Illinois provide appropriate language support services to English language learner students until they no longer need them.

The Department of Justice has reached a settlement that will require GrafTech International Ltd., a major producer of graphite electrodes, to make significant modifications to its supply agreement with ConocoPhillips Company, along with reporting and firewall obligations, in order to proceed with its proposed acquisition of Seadrift Coke LP.

Following a public comment period, the Department of Justice and the Federal Trade Commission (FTC) have made changes to reduce the filing burden and streamline the form parties must file when seeking antitrust clearance of proposed mergers and acquisitions under the Hart-Scott-Rodino (HSR) Act and the Premerger Notification Rules.

The Department of Justice announced today that it will require Verizon and four of the nation’s largest cable companies—Comcast, Time Warner Cable, Bright House Networks and Cox Communications—to make changes to a series of agreements concerning both the sale of bundled wireless and wireline services, and the formation of a technology research joint venture. The department said that, if left unaltered, the agreements would have harmed competition by diminishing the companies’ incentive to compete, resulting in higher prices and lower quality for consumers.

Assistant Attorney General Bill Baer in charge of the Department of Justice’s Antitrust Division issued the following statement today on changes to the division’s carve-out practice regarding corporate plea agreements.

The Justice Department announced today that it intends to file a lawsuit against the State of North Carolina, the North Carolina State Board of Elections, and the Executive Director for the State Board of Elections over recent voting changes made by North Carolina House Bill 589, which was signed into law in August 2013. The United States’ complaint challenges provisions of House Bill 589 under the non-discrimination requirements of Section 2 of the Voting Rights Act.

In testimony delivered before the U.S. Sentencing Commission Thursday, Attorney General Eric Holder endorsed a proposed change to the Federal Sentencing Guidelines that would reserve the harshest penalties for the most serious drug offenders.

High rates of opioid prescribing have contributed to the current U.S. opioid epidemic and the steady increase in overdose deaths across the country. Prescribers wrote an all-time high of 255 million opioid prescriptions in 2012, and though that rate has since declined.

Rousselot, the collagen-based ingredients producer, has revealed a new study which it says provides important answers surrounding the bioavailability of collagen peptides and the modifications they undergo during digestion.

As the industry shifts gears to combat the novel coronavirus, a number of leadership changes have taken place.

April 20, 2020 – With additional changes to the regulations still in administrative review and businesses grappling with pressing COVID-19 issues, a group of nearly 100 advertising trade associations, organizations and companies asked California Attorney General Xavier Becerra (D) to delay the July 1 enforcement date for the California Consumer Protection Act (CCPA). But it […]

npj Microgravity, Published online: 23 April 2020; doi:10.1038/s41526-020-0102-3

Changes to the layout of Valencia's street circuit could be made prior to F1's return next year

Sebastian Vettel said that last minute changes to his car enabled him to take pole position in China