Cleaning the Nano-Texture Glass on Your New iMac – The Mac Observer

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With Apples refresh of the 27-inch iMac, a new option is available that previously was only offered on the Pro Display XDR. You can now ordet your iMac with a nano-texture display. This reduces glare to a bare minimum, but cleaning the nano-texture glass on your new iMac is quite different from other display options.

When you order your 27-inch iMac with nano-texture glass, Apple actually etches the coating into the display at the nanometer level. Apple claims that it reduces glare while preserving contrast, for jaw-dropping image quality. The texturing scatters light as it hits the display, minimizing glare and reducing the undesirable haze and sparkle of the normal matte coatings.

Apple provides a special cleaning cloth for the nano-texture glass. The company says you should never use any other material on the display or you risk damaging the glass. So, how can you clean the nano-texture glass on your new 27-inch iMac if it gets really dirty?

As it turns out, the key to removing difficult smudges is something you probably already have around the house. Take that 70-percent isopropyl alcohol (IPA) solution out of your medicine cabinet. Moisten the cleaning cloth with it, and you can easily wipe away those hard-to-remove smudges.

Once done, you should clean the polishing cloth to remove excess isopropyl alcohol. Just follow these steps:

Should you lose the polishing cloth, or want a spare, you can order a replacement directly from Apple Support.

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Cleaning the Nano-Texture Glass on Your New iMac - The Mac Observer

THE FUTURE JUST ARRIVED: THE ROLE OF BANKS IN A POST-COVID WORLD – Forbes Africa

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THE COVID-19 GLOBAL pandemic has brought forward the future. It has brought about humanitys biggest challenge in a century, to choose between life and livelihoods. In the immediate aftermath of the pandemic, banks have played a supporting role to clients and communities. Standard Chartered announced a commitment of US$1 billion globally to support companies in the health delivery supply chain to combat the COVID-19 pandemic, and an additional US$50 million to assist communities in presence markets, especially Africa.

Generally, banks have entered the COVID crisis much stronger and resilient than they did before the global nancial crisis of 2008. This resilience is owed in part to the raft of regulatory reforms and stronger supervision since 2008, aimed at ensuring adequate capital and liquidity buffers against market-wide stress and also to avoid future taxpayer bailouts. Bank resilience notwithstanding, there is a catch! Well, several!

Many jurisdictions have placed ghting this pandemic at par with war by employing emergency measures and massive scal stimulus packages to provide relief to businesses and households and hopefully to bring the public health crisis under control. Relative to Q1 2020, total lockdown is now perceived as a blunt instrument in responding to the pandemic due to its potential for economic destruction and jeopardizing livelihoods. There is some convergence of views towards recalibrating easing of lockdown alongside placing high emphasis on both corporate and individual responsibility to obey public health recommendations and to observe good hygiene.

As the world emerges from lockdown, the nature and shape of the recovery is an important variable. Whether the recovery is a V-shape, U-shape or a W- shape makes a world of a difference to how banks will respond. We are only beginning to understand and measure the true economic cost of the pandemic, transmitted through lockdown and the deleterious effects on growth and the viability of some sectors. Many companies around the world face the grim prospect of possible collapse, necessitating tough survival decisions on rationalizations, write-downs of asset values and other corporate actions.

National governments need to determine the right balance between restarting economic activity and growth without compromising the capacity of healthcare infrastructure. The ability to do so would in turn inuence the nature and shape of the recovery as well as which companies or sectors can survive the new reality, and which would fail. Then there is digitization. Digitization and the new ways of working will dene who stays competitive, productive and can survive. Almost every sector or industry and even governments will need to invest in digital solutions to future-proof their survival and relevance.

While banks certainly have a critical role to play during and post crisis, the reality is there is no certainty or clarity on how events might unfold. In the meantime, a better understanding of the nature, virulence and measures to conquer the invisible enemy remains elusive, thereby drawing parallels with the expression fog of war. The unfolding challenges and after-effects of the pandemic are not sequential or in any particular order. Neither will the responses be. Responses need to be diligent, intelligent and wise in order to safeguard the future. The pre-COVID bank resilience is a necessary and comforting condition but insufficient for post-COVID recovery unless banks themselves can successfully navigate the fog of this new war to avoid joining the casualty list that is beginning to grow around the world. They will certainly be dealing with elevated credit risk, strains to capital and liquidity and heightened operational and cyber security risk, among others. Livelihoods and economic security look fragile in the short run as the pandemic exposes cracks, ssures and chasms in the existing socio-economic order globally. This systemic stress could precipitate a repurposing of legacy political, economic, social and security arrangements by national governments which could be consequential to businesses recovery and viability, household incomes and social cohesion. For the banks, it becomes a waiting game!

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THE FUTURE JUST ARRIVED: THE ROLE OF BANKS IN A POST-COVID WORLD - Forbes Africa

Emerging T-cell Therapy Treatment for Mesothelioma Shows Significant Tumor Reduction – MesoWatch

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TCR2 Therapeutics announced on July 26 the first positive results for a new T-cell therapy to treat cancer patients, according to the press release on July 26th.

The first five patients treated in phase I of the clinical trial showed mesothelin-expressing tumor regression within six months of treatment. All patients received the same dose of TC-210 TRuC-T cell products. Four patients received lymphodepletion as well and one received TC-210 T cell products without lymphodepletion. Lymphodepletion is when lymphocytes (the cells that fight bacterial and viral infections) are destroyed. Only one patient experienced toxicity as a result of t-cell therapy.

The median reduction range for the sum of the diameters of target lesions was 42 percent.

Garry Menzel, President and Chief Executive Officer of TCR2 Therapeutics expressed gratitude for the results but mentioned that this is just the start of getting T-cell therapy approved for cancer patients.

We are delighted that our very first dose of TC-210 induced consistent tumor regression and clinical benefit in heavily pre-treated cancer patients, Menzel said. There are very few options for patients with solid tumors and those expressing mesothelin represent a significant frontier of unmet medical need. While these are early data requiring further study, we are encouraged by the potential of our TRuC-T cells as we continue to enroll and treat patients with the goal of quickly finding a recommended Phase 2 dose for TC-210.

Phase I of most clinical trials is mainly to assess the safety and efficacy of treatment before increasing dosage in patients. Phase two will use the dosage data from one and test the efficacy in tumor control and regression in about 50 patients, according to TCR2. Phase three is the stage where a treatment seeks approval from the Food and Drug Administration.

Patients in phase two will have a current diagnosis of non-small cell lung cancer, ovarian cancer, malignant pleural/peritoneal mesothelioma, or cholangiocarcinoma and all dosages will reflect the specific cancer type.

While more tests need to be run, Alfonso Quints-Cardama, M.D., Chief Medical Officer of TCR2 Therapeutics, said that this will bring some hope to cancer patients who have failed multiple lines of therapy.

Based on my prior experience working with both TCR-T and CAR-T cells, including the FDA approval of Kymriah, observing consistent clinical benefit in patients at presumably suboptimal T cell doses is quite meaningful, saidQuints-Cardama. These early TC-210 data suggest our approach may overcome the challenges faced by many T cell therapies in the hostile solid tumor microenvironment. Our enrolled patients have failed multiple lines of therapy, including standard chemotherapy, checkpoint inhibitors, and in some cases other mesothelin-directed approaches, in indications where survival has been historically shorter than six months.

According to clinicaltrials.gov, recruiting is still happening for this trial and people can still sign up for phase one.

T-cell therapy is still an emerging treatment for cancers such as mesothelioma. Chimeric antigen receptor T-cell therapy or CAR-T-cell therapy is the only other T-cell treatment that shows promising results. TRuC-T cells would add another form of treatment for mesothelioma, which normally has a poor prognosis.

TCR2 Therapeutics hosted a live webcast and conference call on July 27. The webcast can still be replayed on this webpage until approximately August 26.

For more information on mesothelioma treatment options, visit our webpage on treatments. If you have been diagnosed with mesothelioma or asbestos-related cancer, visit the contact page to talk to an advocate for compensation.

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Emerging T-cell Therapy Treatment for Mesothelioma Shows Significant Tumor Reduction - MesoWatch

Targeted phototherapy study Shows low-risk and effective treatment for Pleural Mesothelioma – MesoWatch

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A new study shows that near-infrared phototherapy could be a promising treatment for individuals with pleural mesothelioma, according to a press release from Nagoya University.

The study, which tested mice and 12 human cell lines with malignant tumors, concluded that near-infrared photoimmunotherapy was a low-risk and effective method of killing cancer cells without harming surrounding cells due to the nature of near-infrared light, according to Kazuhide Sato, a researcher for the Institute for Advanced Research. There are several studies that suggest that near-infrared photoimmunotherapy is an effective cancer treatment, but little or no studies that focus on using this treatment for mesothelioma.

Malignant mesothelioma can be detected with high levels of podoplanin, a protein typically found in cancer cells. This type of treatment targets the expression of podoplanin. First, an anti-podoplanin antibody called NZ-1-IR700 was injected into the mice. This compound binds to podoplanin to both illuminate tumor cells and allow them to absorb light energy. This type of antibody is non-toxic to the body. When Satos team focused a near-infrared light into the chest cavity of mice, they saw the tumor cells rupture.

Sato suggested that further studies look into using a drainage device similar to the one used in patients to drain excess fluid in the lungs. The divide would help diffuse light directly into the chest cavity and cause more effective results.

One possible benefit of this treatment is that healthy cells are less likely to be destroyed. Sato acknowledged that NZ-1 would also bind to podoplanin on lymphatic epithelial cells, but he also suggested using CasMab, an antibody that detects the differences between cancer cells and healthy cells. With or without CasMab, the immunotherapy is regional and can help preserve the lungs. The anti-podoplanin antibodies are also not toxic to cells unless a near-infrared light is on them.

That being said, Sato said that there need to be more studies to ensure that the treatment will not unnecessarily harm the body. This study was partially performed on human cell lines with malignant tumors, but not on human beings.

Near-infrared photoimmunotherapy is a treatment that is perfect for patients with mesothelioma since it is usually localized to the lungs. Sato explained why this therapy works in the chest cavity.

"The lungs and chest cavity contain a large amount of air and are thus very good at effectively transmitting near-infrared light," says Sato. "NIR-PIT is a safe phototherapy option that can target a region of interest. The antibody-IR700 conjugate is also non-toxic to the body in the absence of near-infrared light irradiation. We thus thought that NIR-PIT could be an effective strategy for controlling localized MPM."

Currently, near-infrared photoimmunotherapy is in phase III clinical trials for head and neck cancer treatments for individuals who have failed at least two lines of therapy. The Food and Drug Administration has fast-tracked approval and this could be a possible treatment in the next few years, according to Sato.

Near-infrared light is not visible to the naked eye, but the wavelengths can penetrate the skin up to four inches. The body detects near-infrared light as heat. This type of light does not damage the skin as ultraviolet light would.

Mesothelioma is a rare type of cancer that affects the epithelial lining in the lungs, heart, and other organs. Typically mesothelioma is diagnosed in the late stage with very limited treatment options and a poor prognosis. Near-infrared photoimmunotherapy could bring hope to people with mesothelioma or their families.

For a full copy of the study, go to PubMed.

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Targeted phototherapy study Shows low-risk and effective treatment for Pleural Mesothelioma - MesoWatch

New Study Shows that Surgery Can Improve Prognosis in Patients with Peritoneal Mesothelioma – MesoWatch

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An analysis of mesothelioma patients who underwent cytoreductive surgery concludes that surgical treatment can improve the survival rate of a patient and should be the first choice for early treatment.

A study from the Annals of Surgical Oncology examined the survival rates of over 2000 peritoneal mesothelioma patients from 2003 to 2014. About 51% of patients did not have any kind of surgery to treat their cancer, while 34% received radical surgery according to the study. The patients who received surgery had an average survival rate of about 38.4 months compared to the non-surgery group which was 7.1 months.

Another group was made up of patients who received both radical surgery and chemotherapy. Those patients had a survival rate of about 41.8 months.

The results indicate a major improvement in prognosis. Before the year 2000, the average prognosis of patients with peritoneal mesothelioma was less than a year, according to the study.

The most recent study recommends that more doctors suggest surgery to a newly diagnosed patient to improve life-expectancy should he or she be fit enough to receive it.

Cytoreductive surgery is a lengthy process that involves surgeons inspecting each organ and removing any tumors. Generally, it is a curative treatment in the early stages of mesothelioma and palliative treatment in late stages to improve the quality of life in the terminal stage.

During some surgeries, treatment will also include hyperthermic intraperitoneal chemotherapy, a process where a doctor usually administers chemotherapy drugs directly into the peritoneum to kill cancer cells more. Using cytotoxic drugs post-operatively can prevent more tumors from forming afterward. Furthermore, the direct application of chemotherapy in high concentrations keeps systemic drug levels low, according to one study from the World Journal for Gastrointestinal Oncology.

Hyperthermic intraperitoneal chemotherapy cannot penetrate tumors deeply so surgery is the best preliminary treatment, according to the study.

Cytoreductive surgery can be beneficial for patients who are fit and are in the early stages of peritoneal mesothelioma due to the intensity of the surgery. The procedure lasts about ten hours.

This study referenced another analysis from 2015 where more than 1000 mesothelioma patients were analyzed for their survival rate after surgery and chemotherapy. The results predicted an improved survival rate of about five years and an additional five years if the patient received both surgery and chemotherapy.

According to the report, peritoneal mesothelioma makes up one-fourth of mesothelioma cases worldwide and difficult to detect early. Therefore, cytoreductive surgery is usually a form of palliative treatment. However, the 2015 study did mention that the increase in cytoreductive surgeries with chemotherapy have led to a more-improved prognosis.

If you think you may have been exposed to asbestos at any point in your lifetime, then it is important to get frequent checkups for mesothelioma. Catching peritoneal mesothelioma early can make a major difference in prognosis. You can also talk to someone about receiving compensation for the medical bills.

Peritoneal Mesothelioma is still a difficult form of cancer to treat, but cytoreductive surgery can improve the quality of life for many patients and result in a substantially longer prognosis.

Read the full study here.

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Evolutionary theory of economic decisions | Stanford News – Stanford University News

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Making decisions in the face of uncertainty has never been easy. But the global pandemic has raised the stakes for many previously mundane choices: how to travel, where to get food, when to send kids back to school.

A decision to keep a mixed herd of highly productive goats and drought tolerant camels may help a household survive rare but severe water shortages, even if it doesnt maximize long-term accumulation of wealth. (Image credit: iStock)

Understanding how humans have made high-stakes decisions over evolutionary time may help to explain our choices in the present day including our tendency to veer from the preferences predicted by economic models, according to a new study from scholars at Stanford University and the Santa Fe Institute.

Rather than starting with utility the happiness or value I get out of making my decision now lets think about how the brain was constructed over evolutionary history, said study co-author James Holland Jones, a biological anthropologist at Stanfords School of Earth, Energy & Environmental Sciences (Stanford Earth). The research was published in the journal Evolutionary Human Sciences.

The pairs proposal adds a new perspective to long-running scholarly debates over why practices designed to improve the standard of living among subsistence populations dont take hold, such as the seemingly slow adoption of new farming technologies among poor, small-scale farmers, and more recently, the unwillingness of the poorest poor to adopt microfinance and other development schemes.

There is an inclination to think of the poorest people as being natural entrepreneurs because they have nothing to lose economically, Jones explained. However, the evolutionary logic we employ suggests that the poorest poor have everything to lose and are, in fact, closer to losing it than better-off people. Our model predicts that very poor people would be especially risk-averse.

It also points to the weakness of lean systems in the face of uncommon but severe threats, such as the coronavirus. One of the things were seeing right now is a world that has been optimized for efficiency and is extremely vulnerable to risk, he said. If you scale back organizations to keep them running at a mean level thats high, and you dont have a lot of slack, when a crisis hits youre in trouble.

According to the theory of expected utility, a staple of modern economics, people should always carefully weigh the likelihood of an event along with the prizes or consequences that would accrue from our decision and then choose the option with the highest average payoff. Of course, we rarely calculate these averages in practice, as behavioral economists have long recognized. Yet an assumption that our brains will behave as if we made decisions this way maximizing personal gain at every turn is still baked into many public and economic policies.

We might expect evolutionary systems to mirror markets, with organisms that act rationally out-competing those not behaving rationally, said Jones, an associate professor of Earth system science at Stanford Earth and a senior fellow at the Stanford Woods Institute for the Environment. The catch is that you cant outcompete something if youre extinct.

In addition to influencing policy, business and financial markets, theories of how we make decisions have filtered into popular culture through books like Nudge and Thinking Fast and Slow. However, they tend to deal poorly with choices that humans have faced for the vast majority of their history on Earth namely, those shaped not by market forces, but by environmental variables like temperature or rainfall. In this context, boom times cant compensate for a single lethal bust. Just one bad heat wave, drought, cold snap or flood can leave a household hungry or worse. Variance is what drives you to extinction, Jones said.

As a result, when it comes to preferences that evolve by natural selection, he said, we should expect to see people undervalue long shots that could be profitable, play it safe when things look risky and generally overestimate the likelihood of rare bad outcomes.

On the timescale of evolution, the salient outcome of a decision is how it contributes to fitness, meaning the proportion of the population through time that carries your DNA. Unlike utility, fitness is a measure that multiplies over time. If any generation in your lineage has zero offspring in it, its game over, Jones said. It is a general aversion to zeros that leads to pessimism.

At the same time, fitness plays out over such long timescales that it cant directly influence our behavior. The things that do shape our choices day to day are more like utility in that they can rise and fall without bringing catastrophe. Psychological mechanisms, like satiety or sexual gratification, or something like love of your children, can motivate you in the immediate. They promote fitness in the long run, but they are not the thing actually being maximized over time, he said.

Maximizing fitness leads us to be more pessimistic in our economic decisions than utility models predict. The optimal level of pessimism to promote survival depends on the exact universe the organism occupies, the authors write. For example, hunters targeting rare, big game may stand to bring home more calories if they succeed, but their household could go hungry if they fail. Herders have to weigh not only the productivity of their animals, but also their susceptibility to drought and disease.

Any time where you have to avoid zero, pessimism will pay off, because youd rather leave money on the table than run the risk of going extinct, Jones said.

When social distancing restrictions loosen enough to conduct group experiments, Jones and co-author Michael Price, PhD 15, who studies complex systems as a fellow at the Santa Fe Institute, plan to test their theory with games challenging participants to maximize payoffs that multiply over time or that are hidden but associated with some tangible proxy. By formalizing and eventually testing the theory, the researchers write, they hope to stimulate more work on the possible evolutionary foundations of key results from behavioral economics.

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US Teachers Evolving on Science of Evolution – T.H.E. Journal

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Science Education

Asurvey done in 2007 found that only a third of public high schoolbiology teachers were able to present the subject of evolution in away that satisfied national science experts. And 13 percent ofteachers offered creationism as a "valid scientific alternative"to evolution.

Twelveyears later, teaching practices related to evolution are much better,according to arecently published report.The researchers found "substantial reductions" increationist instruction and a "substantial increase" in thetime that high school teachers devote to human evolution and theevolutionary process in general. The findings were published in thejournal Evolution:Education and Outreach.The researchers came from PennState Universityand the NationalCenter for Science Education.

Whythe uptake in more scientifically astute instruction? The researcherspointed to three changes:

Broad adoption of the Next Generation Science Standards or state standards that borrow heavily from NGSS;

Improvements in pre-service teacher education; and

Changing practices of in-service teachers through improved professional development.

Theproject analyzed data from the "Survey of American ScienceTeachers" which was done between February and May 2019. Whilethe survey included both high school and middle school samples, thisreport relied on high school responses, in particular.

Inboth surveys more than 95 percent of high school biology teachersreported covering evolution to some degree. But there was a 60percent increase in the average number of class hours devoted tohuman evolution in the 2019 survey, an increase from 4.1 hours to 7.7hours.

Also,there was "considerable movement" in how teachers respondedto a question asking them whether they "emphasize the broadconsensus that evolution is a fact, even as scientists disagree aboutthe specific mechanisms through which evolution occurred." Theshare of teachers who disagreed with that statement declined from 22percent to 13 percent, and the percentage who agreed grew from 74percent to 79 percent. Those who "strongly agreed" rosefrom 30 percent to 47 percent.

Theproportion of teachers who reported discussing creationism andintelligent design in classes dropped from 23 percent to 14 percent.And where it was discussed, the researchers pointed out, "someteachers may raise the topic of creationism in the context ofexplaining why it is not scientific." To understand responsesbetter, the survey included two additional questions:

"I emphasize that intelligent design is a valid, scientific alternative to Darwinian explanations for the origin of species"; and

"I emphasize that many reputable scientists view creationism or intelligent design as valid alternatives to Darwinian theory."

Thenumber of teachers disagreeing with the first statement rose from 32percent to 58 percent, a change "largely driven by a sharp dropin the number of teachers who declined to answer this question,"from 53 percent to 29 percent. And there was a big increase "inthe percentage of teachers strongly disagreeing with each statement."The researchers interpreted that to mean that "more teachers areconfident in their acceptance of evolution and rejection ofcreationism."

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US Teachers Evolving on Science of Evolution - T.H.E. Journal

Minnesota Football: The evolution of the slot cornerback – The Daily Gopher

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What is a slot cornerback?

First of all, it is important to establish that under defensive coordinator Joe Rossi the Gopher defense has operated out of a base 4-2-5 formation. That means four defensive linemen, two linebackers, and five defensive backs. Here is a typical look:

Fresno State is in a four wide receiver set, so the Gophers have Coney Durr and Terell Smith as their outside corners, with Chris Williamson in at slot corner on the wide side of the field and safety Jordan Howden providing help over the top. Antoine Winfield Jr. is playing center field as the free safety. Thomas Barber and Kamal Martin are your linebackers, with the more athletic Martin split out to cover the inside receiver on the short side of the hash. The four defensive linemen are Winston DeLattiboudere, Micah Dew-Treadway, Sam Renner, and Carter Coughlin.

Why the 4-2-5? The formation, which is also referred to as the nickel defense, was championed in the 21st century as the answer to the spread offense. TCU head coach Gary Patterson is perhaps the schemes most renowned innovator, having originally utilized it to counter power running teams before evolving it to defend the prolific up-tempo offenses of the Big 12.

I wont get too much into the weeds on the nuances of the 4-2-5 Google Gary Patterson, 4-2-5 for ample supplemental reading but the foundational tenet of the scheme is to flood the field with speed. Athleticism is emphasized over size, even at defensive end, where fleet-footed outside linebackers have been converted into undersized rush ends who can cause problems for slower offensive tackles. Its an aggressive style of defense, one in which the goal is to use mismatches up front to generate pressure, attack gaps, and spill the ball to the perimeter.

The proliferation of the 4-2-5 has seen the role of the nickelback evolve from backup third cornerback to one of the most versatile defenders in modern football. Today, slot corners need be able to blitz the quarterback as pass rushers, step up in the run game as linebackers, and hold their own in coverage as defensive backs. Thats a lot to ask of one player.

Chris Williamson, a transfer from Florida, made the slot corner position his home in his two seasons at Minnesota. He became such a critical part of the Gophers defense that I am more concerned about how Joe Rossi will replace his production than I am about filling the shoes of All-American safety Antoine Winfield Jr.

Seriously.

One of the challenges of playing slot corner is that the slot receiver position is no longer relegated to lesser pass catchers. Teams top targets in the passing game are lining up inside more than ever before. That puts a lot of pressure on slot corners, who are already in a precarious position due to their alignment. They dont have the benefit of using the sideline as leverage or squeezing receivers into the boundary. Slot corners have to play both an inside and outside release. With more area to cover, one false step could spell doom.

Which brings me to a fourth down play against Penn State. On 4th and Goal at the 5, the Nittany Lions lined up in trips formation with three wide receivers on the wide side of the field, and KJ Hamler was in the slot with tight end Pat Freiermuth. Freiermuth ran a slant to try and disrupt the defenders in coverage as Hamler ran a fade to the corner of the end zone. But Williamson reads the route, breaks on Hamlers hip, turns his head, and bats the ball out of bounds.

But as I mentioned previously, pass coverage is only one part of the position.

The slot cornerback also needs to be able to play close to the line of scrimmage, almost functioning as a third linebacker at times. Not every nickel corner who can hold their own in pass coverage is also a reliable tackler. You have to be physical and able to make tackles in space.

Williamson and Justus Harris were relied upon heavily against Georgia Southern last season, both operating out of the slot corner position but playing near the line of scrimmage to help stifle the Eagles option offense in run support. The pair helped limit Georgia Southern to 123 rushing yards (well below their season average of 253.2 yards per game).

Here, Williamson comes in from across the formation to drop the ball carrier for a minimal gain on third down after Thomas Barber whiffs on the tackle.

Last but not least, rushing the passer. On this second down play against Fresno State, Joe Rossi gets creative. DeLattiboudere takes the right tackle upfield and Thomas Barber blitzes, occupying both the right guard and the running back in pass protection. Williamson, sprinting in from his slot corner position, runs through an opening so big that he never even breaks stride en route to the sack. Coughlin is following close behind, ready to take a crack at the quarterback if Williamson cant finish the play, but he requires no such assistance.

Senior Justus Harris is the most experienced slot corner on the roster, but his game action has been limited mostly to special teams. Redshirt freshman Solomon Brown and incoming freshman Jalen Glaze are both expected to compete with Harris at slot corner, but the lack of a clear successor to Williamson is a concern. Williamson was an unsung hero of the defense last season, and replacing his production will be no small task.

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Rapid and robust evolution of collateral sensitivity in Pseudomonas aeruginosa antibiotic-resistant mutants – Science Advances

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INTRODUCTION

Antibiotic effectiveness, currently compromised by the spread of antibiotic resistance (AR), requires not only innovation but also conservation, which may allow for improved use of current antibiotics (1). For this conservation, understanding of the trade-offs associated with AR acquisitionsuch as increased susceptibility to a second drug after use of the first, a phenomenon first described in the 1950s as collateral sensitivity (2)might be particularly relevant. Various studies have been undertaken trying to exploit the evolutionary constraint imposed by collateral sensitivity patterns (3, 4), such as combinatory therapy (5, 6) or alternating collaterally susceptible drug pairs (79).

Despite progress in study of the collateral sensitivity phenomenon, some questions remain to be answered. In particular, there is still only limited information on the evolutionary conservation of collateral sensitivity patterns, not only between different species (10, 11) but also within different members from the same species (1215). In the case of Pseudomonas aeruginosa, some studies have revealed that isolates of this pathogen obtained from patients with chronic infections and treated with distinct antibiotic classes present convergence in their collateral sensitivity phenotypes (9), while others have described major differences in the collateral sensitivity patterns associated with the acquisition of resistance to one antibiotic between replicates of the same strain of P. aeruginosa evolving in parallel (13). These discrepancies may lie in the degree of reproducibility of the evolutionary pathways leading to AR. In this respect, it is known that the type of resistance mutations present in a given genetic background may be restricted because of epistatic interactions (10, 1621) and that the cumulative acquisition of AR mutations in different loci reduces the variety of pathways, leading to AR (22, 23). Since contingency may be relevant not only for AR evolution but also for the acquired collateral sensitivity phenotype (24), knowing the degree of conservation of collateral sensitivity patterns associated with the use of a specific drug in different genetic backgrounds, particularly in the case of mutants already presenting a phenotype of AR, is of special interest.

In a previous study, we observed that P. aeruginosa PA14 populations experimentally evolved in the presence of ceftazidime displayed a robust collateral sensitivity to amikacin (25), as a consequence of selection of large chromosomal deletions upon 1 day of adaptive laboratory evolution (ALE) (25). The chromosomal deletions included hmgA, which encodes an enzyme whose lack of activity leads to the hyperproduction of the brown pigment pyomelanin (26); galU, whose inactivation reduces ceftazidime susceptibility (27); and mexXY, which encodes a multidrug resistance (MDR) efflux pump that contributes to P. aeruginosa intrinsic aminoglycosides resistance (28). Up to 13% of patients with cystic fibrosis (CF) are infected by P. aeruginosa pyomelanin-producing mutants (29) that, in agreement with the phenotype of the mutants selected after ALEs, are also hypersusceptible to aminoglycosides (30). Although the fact that pyomelanin increases resistance to oxidative stress and favors bacterial persistence in chronic lung infections (26) has been considered the most likely explanation for in vivo selection of those mutants, this genetic event has also been reported in different strains of P. aeruginosa subjected to ALE in the presence of other -lactams (3134). Therefore, it remains to be established whether these deletions are selected by the antibiotic treatment or merely represent an adaptation to the lung environment of patients with CF (35), as was previously proposed (34, 35). Even further, it remains to be answered whether these deletions are selected upon ceftazidime treatment in different genetic backgrounds of P. aeruginosa, in particular, in mutants resistant to other antibiotics; or on the contrary, whether the evolutionary robustness of this genetic event, and of its phenotypic effects, is limited. In this study, by constructing a set of resistant mutants previously identified in different ALE experiments of P. aeruginosa PA14 (16, 36) in the presence of antibiotics, and by submitting them, as well as the wild-type PA14 strain, to ALE in the presence of ceftazidime, we have determined the robustness of an early event of ceftazidime resistance evolution that is associated with collateral sensitivity to tobramycin. Further, by the recreation of heterogeneous pyomelanogenic populations belonging to each genetic background and the alternation of tobramycin with ceftazidime, we found that driving evolution toward hypersusceptibility to the first drug is generally feasible, at least in the genetic backgrounds analyzed. This finding supports the possibility of rationally designing treatments based on collateral sensitivity convergence in P. aeruginosa.

As mentioned above, the conservation of collateral sensitivity to a given drug within different genetic backgrounds of a species may be contingent on the degree of conservation of the evolutionary routes toward resistance to the first drug used for selection. We and others have reported the early selection of large chromosomal deletions that contain genes encoding the intrinsic aminoglycosides resistance efflux pump MexXY (28), when ALE experiments in the presence of different -lactams, including ceftazidime (31), piperacillin (32), or meropenem (33, 34), were performed. These data suggest that this genetic event could be one of the first evolutionary steps in the evolution of P. aeruginosa toward -lactam resistance. However, since these studies were limited to a single wild-type genetic background, it remained to be analyzed whether a similar evolutionary pattern could be followed by P. aeruginosa PA14 strains presenting different genetic backgrounds, in particular, different antibiotic-resistant mutants. That being so, collateral sensitivity to aminoglycosides, such as tobramycin, a drug that forms part of usual therapy regimens against P. aeruginosa (37), would also be conserved.

Our first objective was to analyze the evolutionary conservation of ceftazidime resistance evolution in different genetic backgrounds, consisting of mutants derived from different P. aeruginosa PA14 ALEs in the presence of antibiotics (see table S1). It has been recently suggested that AR mutations may associate with either robust or variable collateral sensitivity patterns in different genetic backgrounds, depending on whether they lead to target or regulatory alterations, respectively (10). Taking this hypothesis into account and resorting to previous different in-house ALE experiments (16, 36), we constructed a broad spectrum of strains containing single mutations that affect regulatory proteins (NfxB, ParR, or MexZ), nonregulatory proteins (NuoD or OrfN), or simultaneously containing both types of mutations. The mutant containing mutations in nfxB, phoQ, frr, and pmrB was dubbed MDR6, and the mutant containing mutations in fusA, orfN, pmrB, mexZ, gabP, ptsP, and nuoD was dubbed MDR12 (see table S1 for detailed information). The susceptibility of each mutant to different antibiotics, including those of interest in this studytobramycin and ceftazidimeis shown in Table 1.

MICs 2-fold of the MICs for the wild-type PA14 strain are highlighted in bold. TOB, tobramycin; TGC, tigecycline; CAZ, ceftazidime; CIP, ciprofloxacin; IPM, imipenem.

To determine whether chromosomal deletions containing mexXY would be early selected during P. aeruginosa evolution in presence of ceftazidime in the set of mutants mentioned above, as it was the case in the wild-type strain PA14 (25), four biological replicates of each single (nfxB177, parR87, mexZ43, orfN50, and nuoD184) and multiple mutants (MDR6 and MDR12), and the wild-type PA14 strain, were subjected to ALE in presence or absence of ceftazidime (a total of 64 populations) for 3 days. Upon 1 day of experimental evolution, almost every P. aeruginosa population challenged with antibiotic hyperproduced pyomelanin (27 of 32 populations; Fig. 1A). This result is consistent with the presence of deletions that include hmgA, as those described in our previous study, because pyomelanin accumulation is due to the lack of homogentisate 1,2-dioxygenase activity provided by HmgA (26). Since we had previously determined a cause-effect relationship between the presence of chromosomal deletions containing hmgA and mexXY and the hyperproduction of pyomelanin and hypersusceptibility to aminoglycosides, respectively (25), the susceptibility of each final population to tobramycin was analyzed. All the pyomelanogenic populations obtained after the short-term evolution in presence of ceftazidime were resistant to ceftazidime and hypersusceptible to tobramycin, when compared to the parental strain from which they evolved (Fig. 1B and table S2), even when the mutants were originally less susceptible to tobramycin than the wild-type P. aeruginosa PA14 strain. In particular, tobramycin minimal inhibitory concentration (MIC) was reduced by up to 4-fold in PA14, 2.6-fold in nfxB177, 3-fold in parR87, 7.9-fold in orfN50, 6-fold in mexZ43, 5.3-fold in MDR6, and 10.7-fold in MDR12. Consistent with the linkage between pyomelanin production and deletion of a chromosomal region containing mexXY, nonpyomelanogenic populations (parR87, replicate 1; and nuoD184, all replicates) were not tobramycin hypersusceptible (Fig. 1B and table S2). Further analysis of the results from the experimental evolution study has revealed that six of eight genetic backgrounds presented a significantly (P < 0.01 in all cases) reduced MIC to tobramycin, compared to their parental strains, after the ceftazidime short-term evolution. To further analyze whether chromosomal deletions containing mexXY could be associated with the phenotype of hypersusceptibility, every evolved population, as well as their original parental strain, was genotyped (fig. S1A). A 163-bp polymerase chain reaction (PCR) fragment corresponding to mexXY was detected in every parental strain, as well as in nonpyomelanogenic populations (parR87, replicate 1; and nuoD184, all replicates) and in a pyomelanogenic mixed population (PA14, replicate 2). Consistent with the observed increase in tobramycin susceptibility of most evolved populations (table S2), every pyomelanogenic tobramycin-hypersusceptible population lacked mexXY (fig. S1A). Overall, these results suggest that chromosomal deletions containing mexXY are consistently selected at first steps of ceftazidime resistance evolution in P. aeruginosa, at least in the genetic backgrounds analyzed. To further test whether this observation could be conditioned by the number of replicates used during the assay for each genetic background, 32 replicates of one of the mutants (mexZ43) were subjected to evolution under the same conditions used before, in presence or absence of ceftazidime. We observed that upon 1 day of experimental evolution, a high number of mexZ43 populations challenged with antibiotic hyperproduced pyomelanin (28 of 32 populations; Fig. 1D), just 4 of 32 populations did not become pyomelanogenic (Fig. 1D). Although we are aware that there is a space for unpredictability of ceftazidime resistance evolution in the mutants analyzed, the results here described suggest that alternative genetic evolutionary trajectories in ceftazidime resistance evolution may be limited, and that phenotypic convergence toward collateral sensitivity to tobramycin is robust in P. aeruginosa, even in the case of antibiotic-resistant mutants, as those here analyzed (table S1), after ceftazidime treatment (table S2). However, we are fully aware that all mutants described here derive from P. aeruginosa PA14, and the generalization of our results to other strains will require the analysis of the effect of short-term ceftazidime evolution on a broad and diverse set of clinical strains of P. aeruginosa.

(A) Scheme of the resultant phenotype after the evolution of PA14, single (nfxB177, parR87, mexZ43, orfN50, and nuoD184) and multiple (MDR6 and MDR12) mutants, in the presence of CAZ [left part of (A)] or in the absence of antibiotic [LB; right part of (A)] for 3 days (see Materials and Methods). Pyomelanin hyperproduction was observed in 27 of 32 populations evolved in the presence of CAZ (red-colored cells). (B) Diagram showing convergence toward hypersusceptibility to TOB in the different genetic backgrounds and replicates, analyzed after short-term evolution on CAZ. In all cases, acquisition of a pyomelanogenic phenotype is associated with collateral sensitivity to TOB, irrespective of the genetic background of the evolving strain. MIC values of TOB and CAZ are included in table S2. (C) Isolation of pyomelanogenic clones from each 27 pyomelanogenic population [left part of (A); red-colored cells] obtained in the presence of CAZ. As shown in the figure, the early steps of evolution in presence of CAZ of P. aeruginosa, which lead to collateral sensitivity to TOB and pyomelanin production, are conserved among the different antibiotic-resistant mutants analyzed. (D) Pyomelanogenic phenotype of mexZ43 mutant after 1-day evolution in the presence of CAZ or in the absence of antibiotic (LB). Pyomelanin hyperproduction was observed in 28 of 32 populations evolved on CAZ. Photo credits for (C) and (D): Ins Poveda, Centro Nacional de Biotecnologa. Permission for using these images is not required.

We have shown that ceftazidime selects, in the short term, pyomelanogenic mutants presenting collateral sensitivity to tobramycin in P. aeruginosa, at least in the set of resistant mutants analyzed here (table S1). Nevertheless, there is an extensive heterogeneity within populations of P. aeruginosa in the CF lungs (38), frequently including mutants already presenting a pyomelanogenic phenotype (29, 30), which is usually associated with a reduced susceptibility to ceftazidime. Since this heterogeneity might impair the chances of exploiting the observed tobramycin collateral sensitivity associated with the use of ceftazidime in these heterogeneous populations, we tested the possibility of alternating the antibiotics, first, by using tobramycin and then second, ceftazidime, for reducing the chances of pyomelanogenic heterogeneous populations to escape from the antibiotic challenge.

The strategy would consist of three stages: A first step on tobramycin may force extinction of the ceftazidime-resistant (tobramycin-hypersusceptible) part of the populations, a second step on ceftazidime may drive evolution toward tobramycin hypersusceptibility, and then the extinction of the tobramycin-hypersusceptible cells after switching back to tobramycin (Fig. 2) would be expected. Although this strategy would be also potentially applicable to populations not resistant to ceftazidime, being reduced to a first step on ceftazidime followed by a second one on tobramycin, we decided to analyze its effectiveness in a more complex situation of clinical relevance, as it is the case of heterogeneous pyomelanogenic populations. In particular, diverse populations derived from the set of mutants were analyzed in this work (see table S1). To that end, we isolated a pyomelanogenic clone from each of the 27 pyomelanogenic populations obtained after ceftazidime short-term evolution (Fig. 1A). To note here that although the original clones from which these populations were derived belonged to a diverse set of genetic backgrounds, consisting of antibiotic-resistant mutants that contain mutations in regulatory, nonregulatory, or both types of proteins (Table 1 and table S1), all the isolated clones that hyperproduced pyomelanin (Fig. 1C) were significantly (P < 0.001 in all cases) more susceptible to tobramycin than their respective parental strain (table S3), and lacked mexXY (fig. S1B). Then, we recreated a total of 27 heterogeneous pyomelanogenic populations by mixing each of the 27 pyomelanogenic clones with its respective parental strain in a 1:1 ratio. The heterogeneous populations were dubbed PA14 +1 to +4, nfxB177 +1 to +4, parR87 +2 to +4, mexZ43 +1 to +4, orfN50 +1 to +4, MDR6 +1 to +4, and MDR12 +1 to +4. These populations were first subjected to tobramycin short-term evolution for 3 days (see Materials and Methods), and the capacity of these populations to either evolve toward tobramycin resistance or to go extinct was analyzed (Fig. 3). Since pyomelanogenic clones are less susceptible to ceftazidime (i.e., >256 g/ml in all the parR87 clones; see b data in table S4) than the parental strain from which they evolved (1 g/ml in parR87; see a data in table S4), ceftazidime MIC values were used to verify the extinction of the pyomelanogenic part of each population. After 3 days of evolution in the presence of tobramycin, the ceftazidime MICs for every population (i.e., 1 g/ml in parR87 heterogeneous populations; see the First step (TOB) data in table S4) were close to the ceftazidime MIC for the parental strain [compare First Step (TOB) to the a and b MIC data in table S4]. These data support that the pyomelanogenic part of every population is extinct after the first step of sequential evolution. To further confirm the extinction of the pyomelanogenic clones, the phenotype (green/yellow versus brown color) of 20 clones from each of the 27 populations, a total of 540 clones, were isolated and grown in liquid medium (brown color is poorly appreciated in colonies) to detect any escape of ceftazidime-resistant cells from tobramycin treatment. As shown in Fig. 3B, none of the clones produced pyomelanin, confirming the extinction of the pyomelanogenic part of every population, a fact that was previously hinted by the ceftazidime MIC values of the resultant populations (see table S4). Besides that, each resultant population presented an increased tobramycin MIC, up to 48-fold, depending on the genetic background and replicate (table S4).

Evolution of a heterogeneous population containing a pyomelanogenic (CAZ resistant) subpopulation starts when TOB is added at time zero (t0). In the presence of TOB, there is an extinction of TOB-hypersusceptible pyomelanogenic mutants (red-colored cells) and TOB becomes ineffective (t1). Then, treatment is switched to CAZ, and TOB-resistant cells (contoured gray-colored cells) become TOB-hypersusceptible (t2). Treatment would be switched back to TOB, resulting in the elimination of TOB-hypersusceptible cells (t3). This strategy would also be potentially applicable to initial populations not resistant to CAZ (t1), being reduced to a first step on CAZ (leading to t2), followed by a second step on TOB (resulting in t3).

(A) Diagram showing the evolution of heterogeneous populations (dubbed +1, +2, +3, and +4) containing each parental strain: PA14, nfxB177, parR87, mexZ43, orfN50, MDR6, or MDR12 and four individual pyomelanogenic clones belonging to the same genetic background, during first step of sequential evolution in the presence of TOB (left) and second step in the presence of CAZ (right), for 6 days (see Materials and Methods). In the case of parR87, only three pyomelanogenic clones from independent CAZ-evolved populations (see Fig. 1, A and C) could be isolated. Hypersusceptibility to TOB (contoured and noncontoured red-colored cells) is observed in 23 of 27 populations (see table S4), and pyomelanin production (noncontoured red-colored cells) is observed in 17 of 27 populations. (B) Analysis of extinction of the pyomelanogenic part of the heterogeneous pyomelanogenic populations after a first step of sequential evolution in the presence of TOB [(A) section, left]. The phenotype (color) of 20 clones isolated from each heterogeneous population, after 3 days of TOB evolution, was observed in liquid medium and compared with the color (brown) of each pyomelanogenic parental strain (upper left corner of each plate). In agreement with data shown in table S4, which points to the extinction of pyomelanogenic populations by comparison of CAZ MIC value of each heterogeneous population with the ones of their parental strains and pyomelanogenic clones, the color of the 540 clones analyzed indicated that pyomelanogenic clones were extinct after first step of sequential evolution on TOB. Photo credits for (B): Fernando Sanz-Garca, Centro Nacional de Biotecnologa.

At this point, we specifically focused on the switch from tobramycin to ceftazidime, the second step of the sequential evolution (Fig. 2). Although we had observed conservation of tobramycin collateral sensitivity after evolution in presence of ceftazidime within the analyzed set of mutants of P. aeruginosa (Fig. 1 and table S2), a critical point would be to determine whether this genetic event would also be selected by ceftazidime in the tobramycin-resistant mutants obtained after the first step of evolution of the populations in the presence of tobramycin. Hence, we switched the selective pressure from tobramycin to ceftazidime (see Materials and Methods). As shown in Fig. 3, 17 of 27 populations hyperproduced pyomelanin. A second critical point emerged here: The degree of sensitization to tobramycin of highly resistant mutants to the said antibiotic was uncertain. To determine whether these populations presented an increased susceptibility to tobramycin, as it was observed in the wild-type PA14 background and in most of mutants analyzed in this work (Fig. 1 and table S2), the tobramycin MIC was determined in all the evolved populations. Twenty-three of 27 populations presented an important increase in their sensitivity to tobramycin after switching the selective pressure from tobramycin to ceftazidime, reducing the MIC by up to 128-fold in PA14, 48-fold in nfxB177, 21-fold in parR87, 96-fold in orfN50, 43-fold in mexZ43, 4-fold in MDR6, and 11-fold in MDR12 (Fig. 4 and table S4). This analysis revealed that five of seven heterogeneous pyomelanogenic populations presented a substantially reduced MIC to tobramycin after the second step of sequential evolution on ceftazidime (table S4). These results suggest that it could be possible to exploit the tobramycin collateral sensitivity associated with the use of ceftazidime by switching back selective pressure to tobramycin, although there may be some limitations depending on the genetic background. This was the case of the multiple resistant mutants MDR6 and MDR12, which did not present a relevant reduction in tobramycin MIC after the switch to ceftazidime.

Evolution of TOB MICs (g/ml) of heterogeneous pyomelanogenic populations after sequential evolution on TOB/CAZ (see Fig. 3). Each plot shows the TOB MIC values for a parental strain (PA14, parR87, orfN50, nfxB177, mexZ43, MDR6, or MDR12), indicated as t0 in the x axis, and for four heterogeneous pyomelanogenic populations (represented as black circles) after first evolution on TOB (indicated as TOB in the x axis), followed by second evolution on CAZ (indicated as CAZ in the x axis). Only three heterogeneous pyomelanogenic populations of parR87 were analyzed. TOB MICs decreased after switching from TOB to CAZ by up to 128-fold in PA14, 48-fold in nfxB177, 21-fold in parR87, 96-fold in orfN50, 43-fold in mexZ43, 4-fold in MDR6, and 11-fold in MDR12. MIC values are shown in table S4.

The fact that some populations were hypersusceptible to tobramycin, even without having suffered chromosomal deletions containing mexXY (6 of 27 populations; Fig. 3 and fig. S1C) or being mixed populations (4 of 27 populations; Fig. 3 and fig. S1C), indicates that reciprocal collateral sensitivity between ceftazidime and tobramycin may occur even in the absence of these deletions, a feature that remains to be explored in detail. Overall, our results indicate that this strategy could potentially be applicable from complex situations similar to the ones explored in the current work (heterogeneous pyomelanogenic ceftazidime-resistant populations) to other ones (populations not resistant to ceftazidime).

We have recently described that P. aeruginosa replicate populations subjected to ribosome-targeting antibiotics may present common changes in the susceptibility to other antibiotics, besides those used along selection (16, 36). However, important differences have been described in the collateral sensitivity phenotype among replicate populations of the same P. aeruginosa strain adapted to one antibiotic (13). We have recently reported that a loss-of-function mutant of P. aeruginosa PA14, differing from its parental strain in the activity of just one regulator, not directly linked to AR, presents different patterns of collateral sensitivity and cross-resistance phenotype when it acquires resistance to ribosome-targeting antibiotics (16). Since historical contingency may restrict the evolution of collateral sensitivity and cross-resistance phenotypic outcomes, we wondered whether the populations obtained after tobramycin and ceftazidime sequential evolution, besides presenting a convergent hypersusceptibility to tobramycin, may converge toward the phenotypes of cross-resistance or collateral sensitivity to antibiotics found in other structural families. To address this question, the MICs of a set of antibiotics were determined for the 27 populations. A general pattern of cross-resistance to aztreonam, imipenem, and chloramphenicol, as well as significant collateral sensitivity to fosfomycin, tobramycin, and tetracycline, was observed (P < 0.0001 in all cases) (Fig. 5 and table S5). Since the combination fosfomycin-tobramycin has been found to be synergistic against biofilms of CF P. aeruginosa strains (39) and against P. aeruginosa PAO1 in anaerobic environments (40), we propose that the switch back to tobramycin (Fig. 2) could also be replaced, if necessary, by the combination fosfomycin-tobramycin. To analyze the relative efficacy of the two possible options, we subjected the resultant populations obtained after tobramycin/ceftazidime sequential evolution to ALE in either tobramycin or the combination fosfomycin-tobramycin, applying twice the MIC of each parental strain. We observed that 8 of 27 populations were able to escape from the switch back to tobramycin (orfN50 +3, mexZ43 +2 and +3, MDR6 +1 and +3, and MDR12 +2 to +4). This result agrees with the fact that although most of the populations (23 of 27) presented an important reduction in the tobramycin MIC after sequential evolution on tobramycin and ceftazidime, by up to 128-fold in PA14, 48-fold in nfxB177, 21-fold in parR87, 96-fold in orfN50, 43-fold in mexZ43, 4-fold in MDR6, and 11-fold in MDR12 (Fig. 4), tobramycin MIC values were close to those of parental strains. None of the populations survived the combination fosfomycin-tobramycin, possibly due to the synergistic effect of these antibiotics (39). We therefore propose that use of the fosfomycin-tobramycin combination is more effective than switching back to tobramycin after that treatment.

Collateral sensitivity and cross-resistance to antibiotics from different structural families were analyzed in the 27 populations obtained after sequential evolution. A population is classified as susceptible or resistant when there was an MIC change with respect to the parental strain value. Triangles indicate antibiotics where a predominant change toward resistance (red) or susceptibility (green) with respect to the parental strain was observed. Thickness of the triangle depends on the percentage of conservation of said phenotype. MIC values (g/ml) are included in table S5. AMK, amikacin; ATM, aztreonam; FOF, fosfomycin; ERY, erythromycin; CHL, chloramphenicol; LEV, levofloxacin; TET, tetracycline.

Bacterial evolution is known to be one of the main causes of the current AR problem; but in-depth analysis of this evolution could also help to tackle this issue through the exploitation of the evolutionary trade-offs (as collateral sensitivity) associated with AR acquisition (2, 4). However, the feasibility of this approach requires the collateral sensitivity phenotypes of different resistant mutants to be robust and reproducible (41). In this study, we describe the robustness of collateral sensitivity to tobramycin associated with the short-term use of ceftazidime in an array of P. aeruginosa antibiotic-resistant mutants, chosen on the basis of their differences both in resistance phenotype and in the functions affected by the mutations that they harbor. We propose that the observed evolutionary trade-offs could be exploited for treating both clonal and heterogeneous pyomelanogenic infections. Patients with CF are usually infected by heterogeneous P. aeruginosa populations (38) that include pyomelanogenic mutants (29, 30), which frequently present resistance to -lactams, a feature that could compromise the use of ceftazidime. However, we have found that it is possible to drive the extinction of the pyomelanogenic mutants, first, by using tobramycin and then second, by driving the evolution of the remaining population toward tobramycin hypersusceptibility, using ceftazidime. A bottleneck for the application of this strategy would be the durability over time of tobramycin hypersusceptibility. However, it is important to highlight that the populations obtained after tobramycin/ceftazidime alternation present also collateral sensitivity to fosfomycin. We observed that it is possible to replace the switch back to tobramycin by a fosfomycin-tobramycin combination, which results in higher efficacy. These results point to the possibility of exploiting specific evolutionary trade-offs for tackling the problem of AR. However, we are aware that a detailed analysis determining the degree of conservation of the short-term ceftazidime resistance evolution in a broad and diverse set of clinical strains of P. aeruginosa would be required. Overall, our results and those of others, previously described (41) suggest that the analysis of phenotypic convergence and, in particular, the aspects that deal with the collateral sensitivity of P. aeruginosa AR mutants, is an important step forward in the rational design of therapeutic approaches capable of reducing the AR burden.

Bacteria were grown in LB at 37C, with shaking at 250 rpm in glass tubes. MICs of ceftazidime, aztreonam, imipenem, tobramycin, amikacin, tigecycline, tetracycline, ciprofloxacin, levofloxacin, chloramphenicol, fosfomycin, and erythromycin were determined at 37C in Mueller-Hinton (MH) agar, using E-test strips (MIC Test Strip, Liofilchem).

Four single mutants of P. aeruginosa (nfxB177, parR87, mexZ43, and nuoD184) were constructed by inserting each mutant allele (table S1) by homologous recombination into the wild-type PA14, while the orfN50 single mutant was previously obtained (16). Mutant alleles were obtained by PCR from previous in-house evolved populations (16, 36), leaving approximately 500 bp upstream and downstream the corresponding single-nucleotide polymorphism, using the oligonucleotides described in table S6. PCR products containing Hind III restriction sites were cloned into the Hind IIIdigested and dephosphorylated pEX18Ap vector (42) and then introduced by transformation into the conjugative Escherichia coli S17-1 strain. Subsequently, conjugation and mutant selection were performed, as described elsewhere (42), using carbenicillin (350 g/ml) and 10% sucrose. In all cases, the presence of the mutations was confirmed by Sanger sequencing. To obtain mutants containing multiple mutations, 10 independent resistant clones from end point evolved populations (16) on tobramycin or tigecycline were selected and mutations confirmed by Sanger sequencing. From them, two multiple mutants (table S1), tobramycin or tigecycline resistant, respectively, were chosen. The mutant allele of lasR was replaced by homologous recombination with that of the wild type in the two selected clones, using the above-described strategy and oligonucleotides encompassed in table S6.

Five single mutants, two multiple mutants, and PA14, four replicates of each, were subjected to short-term ALE in presence or absence of ceftazidime, resulting in a total of 64 independent bacterial populations (32 populations grown in presence of ceftazidime and 32 control populations grown without antibiotic). Cultures were grown at 37C and 250 rpm for 3 days. Every day, the cultures were diluted (1/125), adding 8 l of bacteria in 1 ml of fresh LB, either containing or lacking ceftazidime at the concentration that hinders the growth of each P. aeruginosa genetic background under these culture conditions (4 g/ml for PA14, mexZ43, and MDR12; 5 g/ml for nfxB177, orfN50, and MDR6; 3 g/ml for parR87; and 2 g/ml for nuoD184). During the 3 days, the concentration of ceftazidime was maintained. Every replicate population was preserved at 80C at the end of the experimental evolution. In addition, the MIC of the antibiotic used for selection in populations (ceftazidime) and of the one to which ceftazidime evolution gives rise to collateral sensitivity (tobramycin), was determined at 37C in MH agar using E-test strips.

Pyomelanogenic clones were isolated from every individual pyomelanogenic replicate population of each genetic background previously submitted to short-term evolution in the presence of ceftazidime, resulting in a total of 27 pyomelanogenic clones (see above). Overnight bacterial cultures from each pyomelanogenic clone and its parental strain were normalized to an optical density at 600 nm of 4.0 and then mixed in a 1:1 (pyomelanogenic clone:parental strain) ratio, obtaining 27 heterogeneous populations. Cultures were grown at 37C and 250 rpm for 6 days. Every day, during the first 3 days, the cultures were diluted (1/125) in fresh LB containing the tobramycin concentration that hinders the growth of each P. aeruginosa genetic background under these culture conditions (1 g/ml for PA14; 1.5 g/ml for nfxB177, parR87, mexZ43, and MDR6; 4 g/ml for orfN50; and 12 g/ml for MDR12). During the 3 days, the concentration of tobramycin was maintained. At the end of the first step of sequential experimental evolution, every replicate population was preserved at 80C, and the MIC of ceftazidime and tobramycin was determined at 37C in MH agar using E-test strips. The 27 populations were grown, from glycerol stocks, and every day, during the last 3 days, the cultures were diluted (1/125) in fresh LB containing ceftazidime, as described in the above-mentioned section of Material and Methods (see the Short-term ALE in presence of ceftazidime section). Every final population was preserved at 80C at the end of the second step of sequential experimental evolution, and the MIC of tobramycin was determined at 37C in MH agar using E-test strips.

The presence of chromosomal deletions including mexXY in the different genetic backgrounds and their respective evolved populations was analyzed by determining the absence of a 163-bp PCR fragment belonging to mexXY in 2% agarose gel. Primers used for mexXY genotyping are included in table S6.

Data were subjected to pre hoc and post hoc analyses to identify relevant differences, using either analysis of variance (ANOVA), Friedmans, or 2 tests and Dunnetts or Fishers exact test with Hochberg correction, as implemented in R.

Acknowledgments: We thank J. Ramn Valverde, from the Servicio de Computacin Cientfica del CNB, for the helpful discussion and the assistance with the manuscript, and I. Poveda, from the Servicio de Fotografa del CNB, for the photograph support. Funding: This work was supported by the Instituto de Salud Carlos III (grant number RD16/0016/0011), cofinanced by the European Development Regional Fund A Way to Achieve Europe, by grant S2017/BMD-3691 InGEMICS-CM, funded by Comunidad de Madrid (Spain) and European Structural and Investment Funds and by the Spanish Ministry of Economy and Competitivity (BIO2017-83128-R). F.S.-G. is the recipient of a FPU fellowship from MINECO. Author contributions: S.H.-A. participated in the design of the study and performed experimental work, and F.S.-G. performed experimental work. J.L.M. participated in the design of the study. All authors participated in writing the manuscript and approved the submitted version. Competing interests: All authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.

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The evolution of Extinction Rebellion | Environment – The Guardian

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In November 2017, Roger Hallam looked up from his cup of tea in a central London cafe and made a bold prediction. He had been walking me through the principles behind a new air pollution campaign he was organising, which involved small groups of activists blocking some of Londons busiest junctions, when he paused, mid-sentence. Of course, this is just small-scale stuff compared to what is coming, Hallam said. The scale of the ecological crisis is a different thing. It is going to change everything.

The air pollution campaign, Stop Killing Londoners, had yet to gain traction with politicians or the media, but Hallam didnt seem too concerned. He explained that it was partly being used to road-test civil disobedience tactics. Within a year or so we will have thousands of people on the streets, blocking large parts of central London for days on end, he said. Hundreds will be arrested and the government will be forced to sit down and tell the truth about the climate emergency.

It seemed fanciful at the time. Hallam, a charismatic, committed and sometimes divisive figure, was splitting his time between his organic farm in Wales and Kings College London, where he was researching a PhD about political activism. Earlier that year, he had staged a hunger strike, which led Kings College to divest from fossil fuels. Despite this recent victory, and an air of certainty that would become familiar to those who dealt with him in the months to come, there was still little to suggest Hallams predictions would come to pass almost word for word within 18 months. Nothing he had been involved in before had achieved the kind of impact that he was now prophesying. The movement that would carry out these protests, Extinction Rebellion (XR), didnt yet exist.

Extinction Rebellion, which would be founded a few months later, was the brainchild of a small group of activists, academics and friends, including Hallam, and Gail Bradbrook, a similarly unorthodox environmentalist. The daughter of a coal miner from South Elmsall in Yorkshire, Bradbrook studied for a PhD in molecular biophysics before working on various social justice campaigns, such as tax justice and the Occupy movement. But by 2016, disillusioned by how little political progress these groups had made, Bradbrook came to the conclusion that she was missing vital information about how to bring about meaningful change. There was something else that nagged at her, too an awareness of something deeper inside her that needed addressing. (In conversation, Bradbrook moves easily between detailed analysis of the latest climate science and organisational theory to discussions of spirituality. She often holds prayer sessions before meetings.)

In March 2016, Bradbrook travelled to Costa Rica in search of enlightenment. There, in a story that has become part of XR lore, she took a series of powerful psychedelic drugs. The most fucking terrifying thing I have done in my entire life, she told me later. But I was willing to do it because I needed help. Before the hallucinogenic trip began, she offered up a prayer to be shown the codes for social change. Those words would have a key role to play in the emergence of XR.

A few months after she returned to the UK, Bradbrook met Hallam for the first time. Their initial conversation lasted four hours Hallam complained that he had a headache by the end as they discussed the unfolding environmental crisis, the inability of the current system to even recognise the oncoming chaos, and theories about how to transform politics. As Hallam stood up and prepared to leave, he half-jokingly told Bradbrook that the ideas he had laid out about how to transform society were the codes she needed. The hairs on the back of my neck stood up, Bradbrook said later. It was exactly the phrase I had used during my prayer in Costa Rica.

Over the coming months, Bradbrook and Hallam continued their conversations, along with a loose grouping of like-minded people, who eventually formed Rising Up, a network of activists committed to peaceful civil disobedience. It wasnt until April 2018 that, in Bradbrooks home on a hillside overlooking the Cotswold town of Stroud, the idea of Extinction Rebellion was born. At the Stroud meeting, as it has become known, a core group of about 15 long-time campaigners, activists and academics decided that after years of small-scale political campaigns, about everything from fracking to migrant rights, they were, in Bradbrooks words, ready to go for the big one. As another of those present, Simon Bramwell, who was Bradbrooks partner at the time, recalled later: It was [then] that we decided to go for broke. We decided to throw all of our energy and intelligence at something that could change the planet.

But before such grand aims could be achieved, there were more mundane tasks. Agreeing on a name for the new group turned into a 25-step process that went on for weeks. It was an absolute mission, said Clare Farrell, another XR co-founder, who has a background in sustainable fashion and design. When Extinction Rebellion was first suggested, Farrell recalled that there was quite a bit of disquiet, because some people thought it was too harsh. But eventually it won.

According to Bradbrook, the group quickly realised it needed to appeal beyond what she called the leftist echo chamber. The campaign would aim not only to tap into progressive concerns about social justice and equality, but also traditionally conservative themes such as national security and protecting family. XRs goals were boiled down to three demands of the government: to tell the truth about the climate and ecological emergency; to halt biodiversity loss and commit to net zero emissions by 2025; and to follow the lead of a citizens assembly. A presentation titled Heading for Extinction and What to Do About It was developed, and quickly became known as the talk. It starkly outlined the scale of the crisis and the dire consequences of inaction, as well as XRs belief that if enough ordinary people took to the streets for peaceful civil disobedience, then radical change was possible.

Then came the hard work of building a movement from scratch in just a few months. Farrell remembers that those early meetings had a different atmosphere to other campaigns she had been involved in: It felt to me straight away like a group of people who were serious, who were very interested in how the system works and how to change it. She recalled the excitement of working with experts in so many different fields, from design to science, philosophy and politics. It was the breadth of the collaboration that was really fundamental.

Hallam told me at the time that they did not want it to simply be an online phenomenon, which he feared could flare up and then disappear again just as quickly. Instead, XR needed people to go out into their own communities, explain, recruit and then rebel. From the summer of 2018 onwards, volunteers began touring the country, giving the talk to people gathered in libraries and meeting halls, cafes and universities, pubs and churches. These meetings were sometimes tense affairs, as the scale of the climate crisis was laid out. But there was also a sense of relief among many of those present that their fears about the crisis were being acknowledged, and that they were being told there was something they could do.

On 31 October 2018, XR formally launched its campaign with a Declaration of Rebellion outside the Houses of Parliament. It had been billed as a small, symbolic act that would probably attract a couple of hundred people. In a taste of what was to come, more than 1,000 turned up and took a spontaneous decision to occupy the road outside parliament. The groups first big test came the following month, when it held a day-long demonstration aiming to block five bridges across the Thames in central London. On a beautiful winters day, thousands turned out, blocking the bridges and bringing chaos to central London in the biggest act of peaceful civil disobedience seen in London for decades.

Five months later, in April 2019, came the protest that the group had been working towards since its foundation. Thousands of people occupied key sites across London for almost two weeks. More than 1,000 were arrested for what were peaceful, respectful, sometimes joyful, protests. The arrests were a key part of XRs strategy, which drew particular inspiration from the civil rights movements in the US and Gandhis independence struggle in India. By encouraging as many protesters as possible to get themselves arrested, XRs plan devised chiefly by Hallam was to overwhelm the court system, with the aim of winning support and forcing change.

The protests turned XR into a movement of global significance, with scores of XR groups springing up in cities around the world, as well as in towns and cities across the UK. By the end, XRs representatives were sitting down for talks with senior politicians and ministers in the UK. Supporters and funders many of whom had been sceptical before April showered praise and money on the new movement, and in the weeks that followed, the UK parliament and scores of councils around the country declared a climate emergency. XR had changed the conversation around the crisis. Now it had a big question to answer: what next?

Within XR there was elation about the success of Aprils rebellion, as well as exhaustion and uncertainty about the future. A lot of these people had been involved in lots of would-be uprisings that they had hoped would take off but hadnt, so they were wary of pinning too much hope on this, said Daze Aghaji, a 20-year-old student and an early member of XR Youth, the influential youth wing of the movement. And then, when it did all come together, it was a bit like: Oh, OK what do we do now?

Hundreds of new people had come forward to join the movement in the weeks after the April protests, many of them giving up their jobs to dedicate themselves to XR full-time. This influx boosted XRs organisational capacity, but also brought new ideas about what the group should be, and competing theories about how to achieve the change needed to tackle the climate crisis.

Among the key figures to emerge during the April rebellion was Farhana Yamin. Her experience as a former UN lawyer who had helped draft the Paris agreement and worked on many of the key climate treaties of the past three decades enhanced XRs credibility, and she became one of the groups most convincing public advocates. Over lunch at a north London cafe a few months later, she told me she had joined XR after becoming disillusioned with her world of diplomacy, thinktanks and policymakers. I had been working with people who were not telling the truth about where we are, and are in a state of euphoric, egocentric denial, she told me. When she was asked to join XR, she agreed immediately. I saw [coverage of] the October rebellion, when people laid down and got arrested and Greta was there, and I thought: yes, that is what we need, that is what I want to do. It looked beautiful.

Amid all the praise for XRs achievements up to that point, there had also been some significant criticism. From the left, some argued that XRs beyond politics framing fatally overlooked the role that neoliberal capitalism, driven by the right, played in the ecological crisis exploiting people and natural resources, particularly in the global south, for the benefit of a wealthy minority. There was also growing criticism from black and ethnic minority groups, who said XRs tactic of encouraging mass arrests ignored the reality of police racism, and effectively made the protests the preserve of privileged white people.

Meanwhile, from the right, critics branded those involved in Extinction Rebellion variously as middle-class layabouts or drug-addled hippies, dangerous killjoys or extremist anarchists. During the April 2019 protests, Daily Mail headlines described the group as a radical far-Left eco-rabble and detailed how the Extinction Rebellion eco-mob plotted chaos in London from vegetarian caf in leafy market town. (XRs critics have left no cliche unmolested, wrote James Butler in the London Review of Books.)

After April, new recruits also had to be fitted into XRs decentralised structure, which includes an organising hub of circles, each with around a dozen or more individuals who focus on particular themes, from actions to finance, legal support to regenerative culture. On top of this, there are around 485 XR groups spread across more than 70 countries, with about 130 in the UK. There are also scores of affiliated groups based around shared identities, such as XR doctors, XR farmers or XR Muslims. People do not formally join XR, and there is no central membership list. Local groups can plan and carry out their own actions as long as they follow XRs 10 core principles and values, including a commitment to non-violence and focusing on systemic problems rather than blaming and shaming individuals.

Although XRs structure aims, as its website says, to build a participatory, decentralised and inclusive movement, some complain that it allows those with the loudest voices often white, middle-class men to dominate. Others complain of endless meetings, labyrinthine decision-making processes, and the sprawling network of WhatsApp groups the organisation has spawned.

According to Ronan McNern, a veteran of the Occupy movement and a key figure in XRs communications operations, the months after the April rebellion were dominated by internal wrangling. On one side were Hallam and his backers, who were pushing for an escalation in provocative direct action to keep the momentum going. They believed that a relatively small group of people, prepared to keep escalating their disruptive, peaceful, direct action could bring about systemic change quickly especially if the elite in the country are in terminal decline, as Hallam regularly insisted. On the other side were people who argued the good will and moral high ground achieved in April should be used to build a broader movement, within the UK and internationally.

An increasingly important voice in those discussions was XR Youth, a semi-autonomous group that had been created at the start of 2019. I felt pretty early on that as a young person I didnt really fit into main XR, said Aghaji. There was so much love of young people, but not in the right way. I would go to some things and everyone would be like, Do social media! Kids love social media! I was like, this is so sweet, but it is not where we need to be. Another founding member of XR Youth, Nils Agger, 26, who was also one of XRs original co-founders, agreed. I had been feeling for a while that there was something missing, there were just not that many people my own age.

From the start, XR Youth wanted to approach the climate crisis from a more international perspective, to highlight the role of activists in the global south, and to change the perception that this was a struggle for the future, rather than for the present. The message, said Aghaji, was save ourselves rather than save our grandchildren.

As the summer went on and internal disputes about XRs future grew more intense, the conflict came to a head over a proposed action to shut down Heathrow airport. Those in favour, including Hallam, wanted to fly drones within the Heathrow exclusion zone in order to bring the airport to a standstill for days. For others in XR, this represented a counterproductive step-change in the level of civil disobedience. They warned that there could be serious safety concerns, and a real risk that the public goodwill built up in April would be lost.

Towards the end of July, after a series of gruelling discussions, events took an unexpected turn. During a key strategy meeting in Stroud attended by Hallam, Bradbrook and other leading figures, there was a sudden commotion at the door. Aghaji, Agger and half a dozen members of XR youth had turned up, unannounced. Bearing cinnamon rolls with We Love You written in icing on top, they occupied the meeting and read out their three demands, one of which was that the Heathrow plans should not go ahead.

At first they were like: What are you doing coming in here demanding things? recalled Aghaji. But we were like: This is literally what you have taught us to do. Agger said it was a strange feeling, as one of XRs founders, to be occupying an XR meeting: It was us trying to make some kind of intervention in the way XR was working. Heathrow was just the tip of the iceberg. Another member of XR Youth made an emotional speech outlining their concerns. It was very emotionally charged, and very teary, Aghaji said. But, she added, it was a turning point.

A few days later, XR abandoned plans for the Heathrow protest and Bradbrook released a statement on behalf of XR, apologising for failing to address adult privilege within the group, and promising to heal the division and hurts between us. In early September, Hallam and a few others went on to attempt to close down Heathrow anyway, under the banner of a new organisation, Heathrow Pause. The protest, which had already drained so much of XRs energy, fell flat. No planes were stopped or delayed. Several participants were arrested, including Hallam who, after breaching bail conditions by returning to the airport after his initial detention, was subsequently remanded in custody.

In the weeks following the Heathrow protests, hundreds of people who had been arrested during the April rebellion began to appear before the courts. Zo Blackler, a journalist turned activist who oversaw the courts process for XR, said it was a reminder of what the movement was really about: We were presented in the media throughout April as being a young persons movement, or a crusties movement, or a white middle-class movement, or whatever it was that that particular newspaper hated about us. But to her, the hearings showed what those attacks had failed to acknowledge. It was an extraordinary experience to be there having that many people come together and read out their statements about why they had done what they had done. It was very moving. People were crying, and it felt very much as if the judges were listening and responding to what they were hearing. (Earlier this year, one judge said: This is going to be my last Extinction Rebellion trial for a little while. I think they only allow us to do so many of these before our sympathies start to overwhelm us.)

As XR headed towards its October Rebellion labelled internally as the difficult second album the euphoria of April had dissipated, but despite the splits and disputes, the movement had continued to grow, boasting hundreds of groups around the world. Less than 12 months after XRs launch, many were confident that the October rebellion would repeat, or even outdo, its previous success.

One crucial reason XR had risen so far so fast was timing. In the decade before it was founded, the climate movement in the UK had been relatively quiet. Before 2008, a vibrant activist network from Climate Camp, which saw protesters come together to carry out civil disobedience against major carbon emitters, to campaigns against the third runway at Heathrow and Kingsnorth power station had forced the climate crisis up the agenda. And these social movements were at least partially responsible for progress at a political level. In 2008, the Labour government brought in the Climate Change Act, the worlds first long-term, legally binding framework for radically reducing emissions, which committed the UK to an 80% emission reduction by 2050. Among many activists, there was at least some hope that the political class was beginning to grasp the enormity of the challenge ahead.

But when the financial crisis hit in 2008, the focus of politics, and protest, changed, as opposition to austerity became the most urgent concern for campaigning groups from Occupy to UK Uncut, the 2011 student protests to the Peoples Assembly. Paolo Gerbaudo, an academic at Kings College London who studies social movements and had been involved in environmental protests at the Copenhagen climate summit in 2009, said that before the financial crash, the climate emergency felt like the challenge of our time, but afterwards, it had largely slipped off the social and political agenda.

The unfolding ecological crisis, however, had not been put on hold. During the 2010s, evidence of climate breakdown rising temperatures, melting ice, failing harvests continued to mount. As the scientific warnings grew ever more dire, there was a deepening feeling among those who were paying attention that traditional environmental campaigning was failing, and that politicians were not delivering with potentially catastrophic consequences. It was into this context that XR emerged.

In the Autumn of 2018, just as XR launched, UN scientists at the Intergovernmental Panel on Climate Change published an unusually stark report, which warned world leaders that we had only 12 years to make radical changes and avoid the most devastating effects of the climate crisis. According to Blackler, the new movement managed to tap into this feeling of frustration and fear by acknowledging the scale of the crisis, and persuading people that collectively they had the power to change things. We gave people something they could come along to and plug into really easily, she said. XRs message was simple: civil disobedience could actually get results. People suddenly understood that although a few million on a march would achieve nothing, if you could get a comparatively small number to do civil disobedience, you can effect change much, much faster.

As XRs October 2019 rebellion approached, the scientific warnings were becoming, if anything, more dire. The school strike movement, led by Greta Thunberg, had mobilised hundreds of thousands of young people around the world, and public figures such as David Attenborough and Pope Francis were lining up to sound the alarm on the climate and ecological emergency. Everything seemed primed for a repeat of Aprils success.

In reality, it proved impossible to recreate the surprise and novelty that defined the earlier protests. Although XR held more sites in October and said it had more people arrested 1,837, compared with 1,138 in April the protests failed to catch the publics imagination in the same way. Actions that would have been headline news just six months earlier such as hundreds of breastfeeding mothers closing down Google HQ were now seen as par for the course by the media. Other factors also counted against XR. The weather was colder, and the days shorter. Police tactics were harsher and the UK was in the midst of Brexit convulsions.

XR was also criticised for tactical missteps. Early in the morning of 17 October, a number of protesters climbed on top of a tube train at Canning Town station in order to disrupt rush-hour services. Commuters reacted with anger, and scuffles broke out, with one protester being dragged from the roof of the train. The incident only lasted a few minutes, but the footage, which was widely shared, was damaging for XR, and highlighted some of the challenges facing the movement. How do you control what actions go ahead in a decentralised structure? Did the decision to carry out the protest highlight the lack of class and racial diversity within XR? Those who had decided to go ahead with the action did not seem to realise that by targeting the tube at that time of day, the people affected would not be bankers or financiers, but working-class Londoners, many on zero-hours contracts. Many observers also expressed bewilderment at the fact that climate protesters had chosen to target public transport at all.

The UK general election, which took place a couple of months later, was another flashpoint within XR. The Labour party offered a raft of policies to rapidly decarbonise the economy and invest in sustainable, well-paid, unionised jobs: its so-called green industrial revolution. For many people concerned about the climate crisis, this was a cause worth rallying behind. But during the election campaign, XR to the dismay of both Labour party activists and some inside the movement did not mobilise behind Labours climate offer, preferring instead to target all three main parties with hunger strikes and people dressed in bee costumes under the slogan Bee-yond politics. One irate XR figure called me to express her dismay at the group targeting Labour in the run-up to the election. There are some of us who have been in meetings all day really trying to stop this, she said. It is crazy, depressing, but we lost. It is going ahead.

The clash came down to two competing ideas about XRs ultimate purpose. For some, it was a vehicle to transform the political landscape, making it possible for existing institutions to put forward radical environmental policies. For others, the existing structures were incapable of overseeing the transition needed, and had to be overhauled and replaced. Reflecting on these tensions earlier in the year, one insider told me that XR had reached a point where the founders needed to take a step back: It is the classic startup or founder syndrome, where an organisation reaches a point where the founders need to let go and let something else emerge.

At the start of 2020, following Labours general election defeat and the relative letdown of its most recent rebellion, XR found itself at a crossroads. There was concern about the direction of travel, about how well we have managed to decentralise, Bradbrook told me. People [were] getting cross with each other and finger-pointing.

In February, XR released a new strategy document that appeared to move away from occupying and holding sites for days on end, instead emphasising targeted civil disobedience in cities around the country. Plans were announced for a rolling rebellion that would take place in London in May, targeting the underlying causes of the climate and ecological emergency government, media and finance.

Just a few weeks later, as the coronavirus pandemic began sweeping across the UK, the prospect of mass demonstrations receded and XRs plans had to be put on hold. The groups finances had been in a parlous state since October, and they had been hoping for an influx of donations during the May demonstrations. When they were cancelled, XR was forced to scale back its headquarters in London and stop all living allowance payments that had previously supported volunteers in need.

The crisis also had a profound impact on the movement in other ways, according to Blackler. There was a certain point when I remember thinking, Everyone in the country is thinking about their own mortality all the time, and it was such a strange moment, she said. But that is what we need people to be doing to be thinking about these big, existential issues and the mortality of our world.

For many environmentalists, the political response to the devastating pandemic had redrawn the boundaries of the possible. Things that they had repeatedly been told were naive or impossible such as shutting down swathes of the global economy overnight, and trillion-dollar investment packages were suddenly happening. Last year, when XR unveiled its demand for zero carbon by 2025, it was widely criticised as unrealistic. Now, such demands seemed less outlandish. The Overton window has been smashed wide open, Bradbrook told me.

Over the summer, this sense of political possibility grew, as Black Lives Matter protests exploded around the world and millions of people took to the streets to demand fundamental changes to policing and the end of structural racism. According to Blackler, the crises of 2020 have underscored the fact that climate change, structural inequality and racism all have the same root, and that is our global financial system. The protests also refocused attention on the existing criticism of XR from black and ethnic minority groups. On 1 July, XR put out a statement apologising for its previous mistakes: We recognise now that our tactic of arrest has made it easier for people of privilege to participate and that our behaviours and attitudes fed into the system of white supremacy. Were sorry this recognition comes so late.

The BLM protests had created space for XR to have really hard but crucial conversations, said Aghaji. It is a time of maturing. XR fucks up so often because we became way too big too quickly, and none of us foresaw that happening. Now, especially with Covid-19, we have had time to look at ourselves.

In July, XR announced plans for its next rebellion, which will begin on 1 September, when activists plan to peacefully disrupt parliament until the government agrees to debate the groups three demands. In its statement, XR identified an intersection of global crises including climate breakdown, Covid-19, racial injustice and economic inequality as symptoms of a toxic economic system propped up by corrupt politicians that is driving us to extinction. Aghaji saw the statement as further proof that XR was learning to play team sports. She said there were ongoing conversations between XR and multiple movements, and was optimistic about what those new relationships would create during Septembers rebellion.

A few weeks later, in a sign that perhaps a more consensual approach was gaining the upper hand in XR, the group announced that Hallam whom Bradbrook had previously described as XRs biggest asset and worst liability no longer had a formal role with the group. In a statement posted to Facebook on 28 July, Hallam stated that he would be devoting himself to a new direct action organisation/anti-political party, Beyond Politics. I believe immediate high level direct action is a right and duty of all citizens to bring down this genocidal government and those liberal institutions which passively stand by as the greatest crime in human history is taking place, he wrote. Earlier that month, Beyond Politics had targeted the headquarters of several NGOs including Greenpeace and Amnesty, pouring paint over walls and floors and handing over letters demanding they write to their members, urging them to rise up against the government. Meanwhile, XR groups have continued to stage smaller protests, including opposition to HS2 and a demonstration at the British Grand Prix on 2 August during which four protesters were arrested.

Speaking at the beginning of the year, back at her home in Stroud where XR was launched, Bradbrook was reluctant to speculate about the future. There is just arrogance in even having an opinion, she said. But after a pause, she began to imagine a couple of scenarios: one dystopian, one utopian. In the first, humanity destroys itself, and perhaps the universe learns something from that. In the second, a truly global citizens assembly is established, humanity reunites, and we use the sort of energy that has previously been used in a war to go into a rapid healing situation. It is all possible.

She paused again, before adding: I cant think of anything else I would rather do with my life. It is such an honour to try. What else can we all do?

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The evolution of Extinction Rebellion | Environment - The Guardian

The Evolution Of Shea Theodore Is Complete – SinBin.vegas

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Alex Ovechkins office is in the circle on the power play. Patrice Bergerons is at the faceoff dot. Tom Wilsons is in the penalty box. Now Shea Theodore officially staked claim to his new office after an incredible display of passing during the round-robin.

In the 3rd period of last nights round-robin game against the Avalanche, Theodore found himself in a familiar place with the puck. He surveyed the ice, picked out Marchessault, and fired a 100+ foot pass, tape-to-tape to #81 which set up the go-ahead penalty shot.

It was kind of a broken play because Nate was coming out of the box. He was in the zone and he decided to change. Shea had perfect position and it was an unreal pass. -Jonathan Marchessault

Youll have to use your imagination to fill in just how incredible this pass is, but here it is.

That was just one of Theodores many brilliant passes from that very spot. Here are two more from the same game. (There are a few others from the rest of the round-robin, but three gifs in the same article is already a little overwhelming.)

At this point, thats pretty much what we expect from Shea. -Marchessault

But Theodores brilliance throughout the entire round-robin goes well beyond just stretch passes from his newly minted office. Posting four points on two goals and two assists plus an assist on the penalty shot goal, he was Vegas most dangerous offensive weapon behind the $9.5 million man, Mark Stone. Shea averaged 24 minutes a game playing against three of the best lines in the league, he recorded 11 shots on goal during the three-game stretch and was on the ice for seven Vegas goals, tied with William Karlsson for the most of any Golden Knight.

He has the ability to raise his level depending on the moment and how important the game is and I thought you saw that tonight. -Pete DeBoer

Theodores growth as a player with the Golden Knights has been tremendous since he was first acquired at the Expansion Draft. After coming from Anaheim where he competed in 20 playoff games as a rookie, Theodore didnt even make the Golden Knights roster out of the initial training camp. (Theres a strong argument to be made that this was expansion team roster management, but it happened nonetheless.)

He put up 29 points in his first season in Vegas while playing just over 20 minutes a night in a slightly sheltered role. In the playoffs, he was able to tally 10 points in 20 games but had a tough series against Washington which left a sour taste heading into year two. Prior to that season, he held out of camp looking for a long-term deal, which he eventually got, but is starting to look like an incredible bargain for the Golden Knights.

The evolution of his game continued earning a bit more ice time and mustering up 37 points in 79 games in 2018-19. Then in the playoff series against the Sharks, he posted seven assists and eight points and laid the groundwork for what weve seen this year. 46 points total, 18 in 22 games under DeBoer, and an increased defensive role has seen Theodore unseat Nate Schmidt as the Golden Knights clear #1 defenseman.

Tonight, he was elite. I think this guy is going to be in the Norris conversation for years to come here. -DeBoer

Shea was good the first year with us but his game where its at right now, when you talk about top 10 defensemen in the league he has to be up there. Hes improved so much since the beginning of the Golden Knights and his game this year was unbelievable. Hes definitely a big part of our offense. -Marchessault

Theodore has become a household name to any and all Golden Knights fans, but the hockey world as a whole still hasnt quite caught on.

Thats about to change though as the Golden Knights prepare for what they hope to be a deep playoff run. If they go as far as everyone in Vegas hopes, not only is he probably going to become a star in the eyes of the many more hockey fans, but hes got a real shot finding his name inscribed on the Conn Smythe trophy along the way.

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The Evolution Of Shea Theodore Is Complete - SinBin.vegas

Pizza ovens, inflatable tents and USB charging: the evolution of camping – The Guardian

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It used to be considered a low-tech holiday, but while campers who want to get off-grid can still find quiet spots to pitch up and spark up the Trangia, others are unfolding their portable pizza ovens and plugging in their projectors.

Advances in technology, and the rise of glamping and festivals, mean that some holidaymakers are packing a whole range of home comforts alongside their traditional camping gear.

People are making things a bit more bespoke, said Mike Attwooll, product buyer at the camp shop Attwoolls. You go out for the day and back to your base at night and its like home.

Ten years ago electrical hook-ups were unusual, but now they are the norm, he said, as campers look to plug in phones and run equipment they would previously have left behind. He had recently seen photos of someone holding a film night in their tent using a mini-projector, he said.

James Warner Smith of the website Cool Camping said technology had brought modern campers a whole range of choices. Some guys have developed a folding pizza oven that you can take with you. It does rotisserie chicken too, he said. Theres a no-electricity coffee maker, and there are quite few stoves that have USB ports in them so you can charge your phone while you are cooking. One of these, the 149.99 Tegology Tegstove, won investment on Dragons Den.

Attwooll said he had recently been looking at a brochure from the 1960s and many products were the same, but that recent innovations meant better versions were available.

The most important things are what youre sleeping under and what youre sleeping on, he said. Eighty-five per cent of family tents are now inflatable, and campers pump up sturdy beams instead of metal poles.

Iain Geddes, senior technical adviser the Camping and Caravanning Club, said affordable inflatable tents were one of the few big sudden changes in the market. Suddenly I was getting calls from people, like one from a single mum saying can I put one of these up on my own, he said. The fact that the answer was yes has made camping accessible for some who would struggle with traditional tents.

Deflating airbeds have ruined many camping trips, and are now being replaced with a new generation of self-inflating mats which offer comfort and reliability. And there have been changes in sleeping bag technology, which mean those who want a bit more comfort under canvas can get it. Attwooll cites a bag aimed at women which has thicker material at the bottom, and another launched this year by Vango - which comes with a thermal heatpad that can be plugged into a power pack.

Visit any campsite though and you will probably still see at least one bell tent. These have enjoyed a renaissance in recent years, which Warner Smith puts down, in part, to social media. In the Instagram age people have an idea of what things should look like, so they will pitch their bell tent and put up some bunting.

He said changes were being inspired by trips to festivals, and by glamping sites that offer fixed tents, pods and shepherds huts beautifully decorated and kitted out with comfortable beds.

You have a lot of sites where this is a mixture of fixed tents and traditional pitches, and people are seeing the glamping and bringing some of it to their own experience, he said. At the same time there has been the growth of music festivals. Campsite owners have seen the food trucks and some sites will now have them at the weekend.

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Pizza ovens, inflatable tents and USB charging: the evolution of camping - The Guardian

Paper Mario: The Evolution of the RPG Series | CBR – CBR – Comic Book Resources

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Paper Mario is drastically different from the RPG it used to be. Here's how it got from The Thousand-Year Door to The Origami King.

ThePaper Marioseries has come a long way in its 20-year journey and managed to keep fans interested while welcoming new players into the fold. Its latest installment is worlds away from the original game, but Nintendo has managed to transition the series from a beloved RPG into an adventure game, with very few blips along the way.

Here's how Paper Mario became what it is today.

RELATED:Pokmon Twilight Wings' Post-Credits Scene Ties the Miniseries to the Games

Paper Mario was meant to be a sequel toSuper Mario RPG, but things changed when developerSquare Soft left Nintendo for Sony over the lack of storage space on the N64's cartridges. Square maintained certain rights to Super Mario RPG, which forced Nintendo to start over.

Intelligent Systems became the developer and at first,it wasn't sure how to tackle the project.Then Junior Designer Naohiko Aoyama came up with a rough design of 2D characters in a 3D world and submitted it to the team. This led to Paper Mario's ultimate design and name.

Paper Mario, or Paper Mario 64, was designed to be a RPG for kids. The writing was easy to understand and the style was cute, colorful and unique. Combat was turn-based, like many RPGs at the time, but it was simplified. Rather than managing several party members, it focused on Mario to fight, use equipment and level. Partner characters were easy to use and equipment was basic, with progression-based upgrades. It was an all-ages RPG, but it also had secrets, optional bosses and extra systems that made gameplay rewarding even for veteran gamers.

Paper Mario: The Thousand-Year Doorintroduced new gameplay in the form of Mario's paper abilities. He could turn into a paper airplane, roll up or turn sideways to become paper thin. There were new partners and abilities for puzzles. Combat remained the same, but took place on a stage with an audience to help or hinder Mario. The stage visually represented leveling progress, as it got bigger and better at certain levels. This game is frequently named the best in the series.

Related:Waluigi Turns 20: How Luigi's Tennis Rival Became a Cult Favorite

Intelligent Systems wanted the next game to have completely different gameplay. The idea to makeSuper Paper Marioa platformer with the ability to rotate the world came from Ryota Kawade, Paper Mario's Chief Director at the time. This was the first and only time the series completely left the RPG genre.

Thegame took place in the sewers of theMushroom Kingdom as an homage to Warp Pipes. Action-platforming replaced turn-based combat and the game introduced new villains. Like Super Mario Bros. 2, players could choose different characters from the series as they progressed. Partners had abilities, but these were focused more on finding secrets and progressing through worlds.

AlthoughSuper Paper Mario received praise, many fans were disappointed. It was a huge departure from the previous games and there was concern the series wouldn't be a RPG anymore. They were partially correct.

Paper Mario:Sticker Star was originally designed like the RPG games but was scrapped when Miyamoto said it felt too much like TTYD. He told Intelligent Systems that story wasnt important and to not focus on it. This advice was based on a Super Paper Mario survey from Club Nintendo, a discontinued service, where only 1 percent of players said story mattered -- which makes sense, because Super Paper Mariowas a platformer, not a RPG. Because of Miyamoto's advice, Sticker Star was hollow. It remains the least-liked game in the series.

Meanwhile, Nintendo instructed Intelligent Systems to stick with basic Mario characters and avoid unique versions like Koops or the Gourmet Shy-Guy, unless entirely new characters were being created. This is why, in newer Paper Mario games, there are only villains like King Ollie or standard Toads and Koopas, rather than someone like Goombaria.

When Paper Mario: Color Splash was revealed, fans were skeptical because it looked like Sticker Star. However,the world was more engaging with a good story and added comedy. Puzzles were fewer, but better and the Paint Hammer presented a fun progression and challenge for completionists. Unfortunately, Color Splash retained the terrible, irrelevant combat of Sticker Star.Paper Mario was slowly finding its new footing as an adventure game.

RELATED: Nintendo Bans 'Modifying' Mario Villains - Each Game Must Create Its Own

The Origami Kingmanaged to find a good balance for Paper Mario'snew adventure style. Intelligent Systems created new uses for existing characters and crafted a new villain and companion. It also fixed combat by tying it into puzzle-solving and made it relevant to the world. For example, an enemy may be blocking a hidden Toad or access to a puzzle, giving players a reason to battle again.

The Origami King alsotranslated Color Splash's Hammer into confetti to fit the story and built on the completion aspect by adding hidden Toads, blocks and even collectibles. Most importantly, rich storytelling returned.

Paper Mario started out as an RPG because it was an upcoming genre and Nintendo wanted to make a title that anyone could pick up and play. As gaming evolved, adventure games became the simpler genre, so Intelligent Systems reinvented the series. It changed everything about Paper Mario and while its unlikelythere will be a return to the RPG format,Paper Mario is finding its new footing.

KEEP READING: Paper Mario: Tips, Tricks & Strategies for New Origami King Players

Dungeons & Dragons 5e: How to Run a Dark Souls-Themed Campaign

An avid gamer with a poochy sidekick living just outside of Ottawa, Ontario. When not gaming she dons capes for writing, creating digital art, and sometimes game development. Games she's worked on can even be found on the app stores. Also loves metal music, horror movies, and a good book.

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New Marvel Video Shows Iron Mans Suit Evolution Over The Years – Screen Rant

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A newly-released promotional video for the Marvel Cinematic Universe tracks Iron Man's suit evolution from Iron Man's Mark I to Endgame's Mark 85.

Iron Man's (Robert Downey Jr.) suit evolution in the Marvel Cinematic Universe over the years is shown in a brand new video. The franchise's inaugural hero used his self-made armor to carry out his responsibilities as an Avenger. Thanks to his penchant for improving his tech, he continued to tinker on his suitsthroughout his stint as a superhero with his final one his most futuristic creation.

Tony Stark's first suit, however, wasn't madewith the intention for it to be used in combat. It's simply a means for him to escape his captivity in Afghanistan in the first Iron Man film. He created it with limited resources and poor conditions, but with the help of brilliant surgeon Ho Yinsen (Shaun Toub), who saved his life. Upon his return home, the armor becamea reminder that he could use his technology himself to ensure that it's being utilized for the better instead of handing it off to other people who might do more harm than good with it. Since then, the genius billionaire had regularly put on his Iron Man suit for various missions until his death in Avengers: Endgame.

Related:Every Iron Man Replacement Marvel Has Already Introduced

A new promotional clip released by Disney+ tracks the evolution of Iron Man suits starting from the bulky and make-shift Mark I from 2008's Iron Man to the sleek and high-tech Mark 85 from Endgame. The video was rolled out as part of marketing all the Iron Man films,other MCU films starring Tony Stark that are now available in the streaming platform. Check out the clip below:

The video also provides pertinent details for each suit, including its name and where fans have seen them. While each MCU hero always debuts a slightly different suit in each film to help with the sales of toys and other pieces of tie-in merchandise, Iron Man is one of the few characterswhose countless armor upgradesalso made sense in terms of story. Since Stark created his suits without the need for any help from other people, he's able to consistently improve them. More often than not, the upgrades stemmed from previously-exposed weaknesses in his other suits. In some instances, Marvel Studios even used him to explain how his fellow Avengers got better costumes or weaponry.

Aside from primarily wearing the Iron Man suit, Stark's genius also led him to create armors for the people closest to him such as his best friend Rhodey (Don Cheadle) and wife Pepper Potts (Gwyneth Paltrow), transforming them to become War Machine and Rescue respectively. Now that he's gone in the Marvel Cinematic Universe, it's curious how those armors can get upgrades moving forward - especially War Machine's who's still expected to carry on his superhero duties moving forward.

More:Why Avengers: Age of Ultron Doesn't Fit In The MCU Phase 2

Source: Disney+

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John Wick 5: Keanu Reeves Already Revealed The Franchise's Best Ending Idea

Accidental geek who is perpetually curious, Ana rekindled her love for writing several years back and married it with everything pop culture. The result is a passionate young writer who could ramble (and of course, pen) about films and series multiple hours a day. She has a soft spot for The Lion King, old songs, and home design; is currently obsessed with old sitcoms (The Golden Girls!); and won't dare watch any horror films although shes (ironically) dying to see one. Though a bit late to the party and was an actual Force non-believer, she now finds the Star Wars franchise quite fascinating (fun fact: it was a crazy Jar Jar Binks/Sith theory that drew her in).

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New Marvel Video Shows Iron Mans Suit Evolution Over The Years - Screen Rant

Pokmon Go Enigma Week: How to get Elgyem and evolve it into Beheeyem – HITC – Football, Gaming, Movies, TV, Music

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Pokmon

Pokmon Go just had its August community day which involved a shiny Magikarp, but Enigma Week continues for a further couple of days. While Enigma Week continues to last, you have better odds at capturing an Elgyem and evolving into Beheeyem. Here youll discover everything you must know about how to get Elgyem and complete its evolution into Beheeyem.

Although July was host to a bunch of tasks and rewards, as well as Pokmon Go Fest itself, August still remains an exciting month for the community as there is a global makeup event for Go Fest right around the corner. Not to mention theres also Unova Week which will be commencing very soon.

But, before Unova Week commences, here youll discover how to get Elgyem and evolve it into Beheeyem before Enigma Week finishes.

You have the chance to catch Elgyem in Pokmon Go while Enigma Week lasts.

Enigma Week finishes on August 14th at 13:00 PT, 16:00 ET, and 21:00 BST, and its your best chance to get and catch an Elgyem in Pokmon Go.

This is because its spawn rate in the wild is increased along with the following creatures:

In addition to being able to encounter Elgyem in the wild, Enigma Week also provides the chance for it to hatch from 7km eggs.

You need 50 candy to evolve Elgyem into Beheeyem in Pokmon Go.

That is how you complete the evolution of Elgyem into Beheeyem, and Pokmon Gos Enigma Week is again your best opportunity.

Aside from catching and evolving the aforementioned creature, theres still plenty more to do during Enigma Week such as catching shiny Staryu and battling Deoxys Normal Forme.

In other news, Fall Guys: How do you change your name on PC for Steam?

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Pokmon Go Enigma Week: How to get Elgyem and evolve it into Beheeyem - HITC - Football, Gaming, Movies, TV, Music

Evolution of Singapore’s ILS infrastructure to attract new sponsors – Artemis.bm

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As Singapore evolves as an insurance-linked securities (ILS) domicile and enables regulatory, legal and tax changes, the ability of the region to facilitate ILS in all forms will attract a growing number of new sponsors, according to industry experts.Singapore has ambitions to become a leading hub for ILS business in Asia, a goal underpinned by the extension of the Monetary Authority of Singapores (MAS) ILS grant scheme to the end of 2022.

The extension of the scheme, which was revealed at the virtually held Artemis ILS Asia 2020 conference, means that 100% of certain upfront issuance costs of catastrophe bonds in Singapore are now funded by the scheme until December 31st, 2022.

The scheme was launched in 2018 to encourage ILS issuance in Singapore and has been well received by the industry, with take-up levels promising.

But, two years after its launch, is the region now ready to expand its remit and embrace ILS in all its forms?

According to Simon Goh, Partner, Head of Insurance & Reinsurance Practice, Rajah & Tann Singapore LLP, Singapore is ready and, this is a critical step in the development of the region as an ILS domicile in Asia.

I think the first step now is to extend the grant, the next step is ILS instruments, said Goh. The next evolution really is to expand into those other areas where we have seen quite a lot of interest.

Gohs comments came during the first panel session of ILS Asia 2020, which also featured Elean Chin, Deputy Director & Head of Insurance, Funds & Infrastructure Finance Division at the MAS.

In agreement with Goh, she told the audience that as part of its next phase of growth, we certainly want to diversify the ILS use cases that Singapore can support.

However, in order to facilitate a diverse range of instruments, including collateralised reinsurance, sidecars, and industry loss warranties (ILWs) alongside different types of risks beyond nat cat, such as cyber, terrorism, and operational risks, Chin highlighted the need for underlying infrastructure and frameworks that enable and support this evolution.

I think Simon touched on one aspect, which is the regulatory framework, but, equally we are looking at the corporate and legal structures as well, including whether or not Singapore should be putting in place a protected cell company structure which we have mentioned previously, to support multiple issuances. Because it actually supports the efficient segregation of assets and liabilities in a very robust way, said Chin.

Alongside both the regulatory and legal structure, the tax structure is another aspect that needs to be looked at in Singapore because currently, the regions tax incentives are more geared towards debt based securities like cat bonds, explained Chin.

Adding, I think this is all part of the ILS 2.0 that my team is doing.

In response to Chins comments, panellist Cory Anger, Global Head of ILS Origination and Structuring at GC Securities, a division of reinsurance broker Guy Carpenter, welcomed the fact that the domicile is exploring changes to its underlying infrastructure in order to be more supportive.

Im encouraged to hear about the regulatory and the tax changes that are being discussed because some of those clients (potential sponsors) very much want to be able to execute, not necessarily in cat bond form, but to be able to do it in different formats, whether its sidecars etc.

So, I definitely think that youll see folks that have never been present in the ILS market in any fashion, come to the market once some of these changes are enabled from them, said Anger.

Adding, And, by virtue of the regulators focusing on becoming comfortable themselves with ILS, is a signal to the market that ILS is an acceptable tool. So, that does change the nature of conversations.

Again, insurance penetration and growing coverage needs are also catalysts along with pricing, but I think all of these together we will definitely see a number of opportunities come through. But, they do take time.

The full video of the panel discussion is embedded below and can also be viewed, along with previous Artemis Live video interviews, over on our YouTube channel.

You can also listen in audio to these interviews by subscribing to the Artemis Live podcast here.

All of our Artemis Live video interviews have a focus on reinsurance, ILS and the efficiency of risk transfer and can be accessed directly from our YouTube Channel.

You can also listen in audio to these interviews by subscribing to the Artemis Live podcast here.

Were offering sponsorship opportunities for our broadcasts, including pre-roll adverts or simply sponsor the entire series, as well as facilitating one-off sponsored webinars and virtual events including roundtables, to help companies access our market-leading audience. Please get in touch to discuss.

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The Evolution Of The Nissan/Datsun GT-R In Pictures | HotCars – HotCars

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The Nissan Skyline became one of the most popular cars in the world, all thanks to race wins and video games. Here's the GTR's evolution.

Today there are few cars as highly touted, as well know, or as lauded as the Nissan GT-R. It's an icon for so many reasons and it's been around since before Ford graced us with the mighty Mustang or Chevrolet shouted back with their killer Camaro. In fact, it's been branded under three different parent company names. It's featured four-doors as well as coupes, and today it stands alone as the only Japanese car to have the ravenous following that it does today.

Often nicknamed Godzilla, it's fascinating to review how this giant killer was born, how it grew up, and why it's earned that nomenclature. When you consider the amazing sports cars from the island of Japan, none come close to garnering the same attention. The Supra has long since lost much of the luster it gained after The Fast & Furious brought it back into the limelight. Old rivals like the 3000GT VR-4 have long since died. The legacy of the Skyline and its bloodline is one of potent sticking power. Let's dig in.

In 1957 a company called Prince debuted the Skyline as an economy car of sorts. In many ways, you can see the design influence of the same years Chevrolet Bel Air.

It was powered by a 60 horsepower 1.5-liter engine and only really turned into a performance car in 1964 with the introduction of the GT Skyline.

In 1969 the car was finally a Nissan branded product and models like this GT-R were highly touted as performance vehicles in its home country.

It housed a six-cylinder engine that produced 160 horsepower. That power was good enough to help it win 52 races in just three years of racing.

RELATED:15 Of The Sickest Skylines Were Obsessed With

In 1972 the Skyline became known as the Datsun K-Series. In fact, the K-Series was the car that after a short departure brought back the iconic round taillights found on every version of the car that followed it.

While some GT-Rs were created in the K-Series, the oil crisis saw that version killed off not long after it debuted.

The R30 was the first really modernized Skyline that started to shape the future of the vehicle. From the moment it arrived in 1981 Nissan worked at improving it each successive year and it showed.

Each year it became more powerful, more capable, and with models like the 2000RS lighter. All of this progress resulted in a return of the GT-R model to the lineup and serious racing capabilities that are still lauded to this very day in the region.

When the R31 Skyline arrived in 1986 with a slightly modernized shape and more features it started to garner the reputation of a truly all-around sports car. Comfortable, fast, and attractive.

It was also the first Skyline to be imbued with the power that came from the RB Redtop Nissan engine family. The continued to build on the advancements of the R30, adding a bigger turbocharger and four-wheel-steering to some models.

Then the R32 showed up and threw down the gauntlet. After years of slowly crafting an excellent sports car, in 1989, this version destroyed the competition. It's quite simply one of the most dominant racing platforms ever seen in motorsports.

In four years of racing during one series the Skyline entered 29 races and won EVERY SINGLE ONE OF THEM. The road car took many queues from the race car and the legend became unstoppable.

RELATED:These Sporty Japanese Cars Left The Competition In The Dust

The R33 is by many accounts, the ugliest of the modern Skylines. It's a fine car though. You'll notice by this point how, much like Porsche, Nissan wasn't making wild changes and dramatically different cars as much as they were taking all the lessons learned from the previous generation and making small tweaks to add performance.

The R33 was no change in that successful formula. To this point, it was the safest Skyline ever produced as well and the 6-cylinder engine became standard on all models.

Here's the one everyone wants,including me. The R34 Skyline is the car that so many of us used in Gran Turismo to decimate all as I've heard it put. It was available with a twin-turbo six-cylinder that could handle more power than Jeff Bezos has and looked like it was going fast even when it wasn't. The internal technology was incredible too.

Nissan employed a screen that told drivers far more vital info about their cars than any had before. Quite simply, this is the one everyone remembers even if they've never seen one in person.

As a person living in the USA, I can say from experience that this was an answer to many people's demands. After decades in countries all across the globe save for the United States, Nissan finally built one that could be sold here. It's been a staple of the track day, autocross, street racing scene ever since.

This most modern GT-R wasn't branded as a Skyline, a marketing misstep perhaps but it hasn't hampered sales. Well known as the cheapest ticket to ludicrous speeds here, the R35 is so bespoke and well-built that now, more than a decade later it's barely changed and is still highly regarded. That can't be said for it's aging baby brother the 370Z.

The GT-R 50 is a wild collaboration between Nissan and Italdesign. Intended to be a totally bespoke and super future version of the GT-R it's pricey at more than $1,000,000. That price comes with ample power to the tune of 710bhp. The trouble seems to be that Nissan can't find buyers.

For such a rare and bespoke vehicle normally these things are snatched up before they're even revealed but after more than a year now, there are still some available should you be in the market.

NEXT:Nissan Finally Reveals GT-R 50 By Italdesign In Track Photos

Next 10 Real Supercars That Are Actually Dirt Cheap Now

Stephen Rivers is a 6'6 racing junkie who's owned 2 BMWs, 3 MINIs and still owns a Rallycross tuned WRX. His dream car is a TVR Cerbera, couple that with his history and he's clearly a masochist.

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The Evolution Of The Nissan/Datsun GT-R In Pictures | HotCars - HotCars

Mixed Martial Arts Equipment Market Overview, Experiments, Evolution, Manufacturers and Forecast until 2026 – Kentucky Journal 24

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The term mixed martial art scan be defined as a combat sport which involves both striking and grappling, using techniques from martial arts and other combat sports. The sport has rivalled professional wrestling and boxing and has witnessed an increased popularity with a pay per view business model. Various styles have been developed across regions with their uniqueness. Various MMA equipment such as mouthpiece, gloves, hand wraps, head gear, shin guards, etc., are used during combat. Global Mixed martial arts Market is expected to grow in the forecasted period, in 2018 market size of theMixed martial artswas XX million and in 2026 is expected to reach at XX million with growing CAGR of XX%.

Get Access to sample pages @https://www.trendsmarketresearch.com/report/sample/3334

Market Dynamics:

Factors such as the increased popularity of the mixed martial arts organizations such as the ultimate fighting championship (UFC) on social media channels and existence of strict regulations and classifications that offers non-biased rules in the execution of mixed martial arts inspire many people to take up mixed martial arts. This will lead to the rise in the demand for mixed martial arts equipment, driving the markets growth prospects.The technological innovations are one of the key trends that will contribute to the markets growth in the forthcoming years. Also, many vendors in the global mixed martial arts equipment market have introduced products that are fitted with tracking sensors.

Market Players:

Adidas AG, Century LLC, Everlast Worldwide, Inc., Hayabusa Fightwear Inc., Combat Sports Inc., Twins Special Co. LTD., Fairtex, King Professional, Title Boxing, LLC and Ringside, Inc.are some of the prominent players in the Mixed martial arts Market.

Market Segmentation:

The mixed martial arts market is classified by product type, sales channel and region. On the basis of product type the market is segmented into gloves, ankle/ knee/ elbow guard, punching bags, hand wraps, shin guard, mouth guard, head gear and others.Further, based on sales channel the market is classified into independent sports outlet, franchised sports outlet, modern trade channels, direct to customer institutional channel, direct to customer online channel and third party online channel.

North America region shows higher market attractiveness index and is considered to be the most lucrative region for the global mixed martial arts equipment market. The rise of the sport in this region along with technological advancements and higher disposable incomes of people have contributed to the growth of the market in this region.

Market segmented on the basis ofsales channel:

Independent Sports Outlet Franchised Sports Outlet Modern Trade Channels Direct to Customer Institutional Channel Direct to Customer Online Channel Third Party Online Channel

Market segmented on the basis of product type:

Gloves Ankle/ Knee/ Elbow Guard Punching Bags Hand Wraps Shin Guard Mouth Guard Head Gear Others

Market segmented on the basis of region:

North America US Canada Mexico Europe UK Germany France Rest of Europe Asia-Pacific China Japan India Australia Rest of Asia-Pacific Latin America Brazil Rest of Latin America Middle East and Africa (MEA) South Africa Saudi Arabia Rest of MEA

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Mixed Martial Arts Equipment Market Overview, Experiments, Evolution, Manufacturers and Forecast until 2026 - Kentucky Journal 24

Equity, Inclusion And the Evolution Of Venture Finance – Entrepreneur

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Is the global private capital landscape ready to finally begin shifting from its historical pale and male dominance?

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August7, 20206 min read

Opinions expressed by Entrepreneur contributors are their own.

Commitments to gender and racial representation in financial markets have been escalating over the past several years, fromGoldman Sachsto theInternational Finance Corporation. And it's no wonder, research from McKinsey & Co. has indicated that companies in the top quartiles for ethnic diversityare 35 per cent more likely to generate higher than average profits. Similarly, companies with the highest gender and ethnic diversity in managementhad innovation revenue that was on average 19 per cent higherthan counterparts with below-average diversity in leadership. Meanwhile, Fortune 100 companies with the highest womens representation in leadershipgenerated 53 per cent higher returns on equity.

In line with this shift in the global financial landscape, gender lens investing has leapt fromfunds managing $2.2 billion in 2018, to 138 funds managing nearly $5 billion by 2020. More recently, in the aftermath of the George Floyd lynching and a global Black Lives Matter uprising, many funds have likewise pledged to begin backing greater shares of Black founders and Black and minority ethnic GP's. In fact, racial justice index funds and ETFs are gaining increased momentum and investor attention.

Nonetheless, shifting to a more gender and racially inclusive investment ecosystem may mean shifting the modus operandi, especially when it comes to funding underrepresented entrepreneurs.

Data fromvarious acceleratorsand government research suggest that women-led ventures in emerging markets get stuck at earlier stages than their male counterparts. This is in spite of regions such as sub-Saharan Africa recording more women than men starting up businesses. This discrepancy stems from various factors. For example, the gender pay gap limits womens availability to bootstrap for as long and as far. Women are also shoulderingan average of three times greater childcare and household labor burdens, which further diminishes availability of time and capital for their businesses. In a post-COVID world, this is exacerbated by the fact thatwomen are facing greater income lossesdue to factors that include womens sector representation in the hardest-hit sectors, as well as workplace unconscious bias. And when women go on to accelerators and incubators, they reap far lesser gains in investment that would propel them beyond seed and early-stage. Initial IFC and Village Capital research shows male-led companies raise nearlythree times more equitythan female-led companies graduating from such programmes.

Initial research is showing that while women-led companies are less likely to raise capital,they perform better and deliver more than male-led counterparts when they do receive it.

As each investors strategy will be heavily influenced by key factors, including but not limited to sector, stage, market, and LP requirements, there is no one-size-fits-all tactic that can be applied across the board to make all fund managers and their portfolios gender-equitable and racially-representative. However, there are few core action areas.

One piece of the puzzle requiring attention is the lack of diversity in investors and investment committees where the majority of venture capitalists are male and white. In a world where75 per cent of white Americans admit to having no social relationships with Black Americans, there are obvious pipeline limitations for the majority of venture capitalists seeking more diverse entrepreneurs. Perhaps the most important long-term step to building a demographically representative portfolio is building a more diverse team and a gender and racially representative investment committee.

Another more immediate strategy is to leverage partnerships that build pipelines and can also provide market, cultural and/or consumer insight. For some VC firms this may simply mean partnering with local fund managers in emerging markets to better identify and evaluate opportunities that can unlock high returns. This is what US-based VC fund Enygma Ventures did through partnering with Africa Trust Group to drive investment in women-led businesses in Southern Africa. Such partnerships are particularly valuable with cross-border and cross-cultural investment into diverse founders, as local partners tend to be better able to spot and understand the non-obvious opportunities available within certain contexts and to surmount typical language and cultural barriers.

There is an approximated$3+ trillion global funding gapfor businesses that dont meet venture capital requirements nor have enough assets to meet collateral requirements of traditional lending. For some companies this may simply mean short-term debt. This is particularly interesting in light of the unique obligations of women entrepreneurs. For example, in some emerging markets, women are less eager to take equity investment, because of the social expectations regarding their role in the family and household. Equity investments require entrepreneurs to forfeit more control over their business strategy and operations to their investors - the consequences of potential loss of control and ensuring compliance with equity investors is likely even more intimidating to the women entrepreneurs already struggling with balancing venture growth and family /household obligations.

In attempting to create a more equitable ecosystem, there's an opportunity for investors to adapt and/or innovate financial products to better serve entrepreneurs, particularly women entrepreneurs. In order to better serve women-led companies, Africa Trust Group is exploring drawdown accounts, which still give investors visibility yet are completely fit-for-purpose. At the same time, revenue sharing, non-dilutive revenue-based financing and quasi-equity models can be an attractive option for early-stage B2B companies. More than serve as benchmarks, these examples serve as beacons calling attention to the large gaps in early stage capital ecosystems.

All of the action areas mentioned must be driven, informed by data and analysis, and as such, will be iterated over time. Acknowledging gender and racial inequities in private capital markets has been an overdue awakening, as theres such significant evidence of the handsome upsides to creating a more representative financial system. More responsive and diversified private capital markets will end up being more profitable and sustainable for investors and more impactful in generating inclusive growth.

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Equity, Inclusion And the Evolution Of Venture Finance - Entrepreneur

FDA Publishes Report on Nanotechnology Over a Decade of Progress and Innovation, Will Hold Webinar on August 13 – JD Supra

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The U.S. Food and Drug Administration (FDA) published a report entitled Nanotechnology Over a Decade of Progress and Innovation that highlights FDAs advancements in the field of nanotechnology since it released its last report in 2007. The report also reviews FDAs role in advancing the public health through its regulation of products within its jurisdiction that involve the application of nanotechnology. According to the report, FDA will rely on a combination of horizon-scanning activities to stay abreast of new developments and product applications, including by:

In addition, FDA states that its Emerging Sciences Working Group, a cross-agency, science-based forum established in 2016, continues to identify science and technology trends of relevance to FDAs regulatory responsibilities, including those for nanotechnology products. FDA notes that its science-based, product-focused regulatory framework is sufficiently flexible and robust to help ensure product safety (and effectiveness, as applicable) while supporting innovation for the development of beneficial nanotechnology products.

On August 13, 2020, FDA will hold a webinar to present the report. The webinar will include the basics of nanotechnology and will highlight the facilities, regulatory science research, guidance documents, standards, domestic and international collaborations, and emerging challenges in regulatory science. The speaker will be Anil K. Patri, Ph.D., Chair, Nanotechnology Task Force, Director, Nanocore, National Center for Toxicological Research (NCTR)/FDA.

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FDA Publishes Report on Nanotechnology Over a Decade of Progress and Innovation, Will Hold Webinar on August 13 - JD Supra