Stu Heritage: Im raising my sons to be the very best feminists – Marie Claire UK

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Author and Guardian columnist Stuart Heritage knows all too well how toxic masculinity has had a detrimental effect on womens fight for gender equality, freedom from sexual harassment and womens rights. So hes laying down the ground rules for his sons as they grow up in a post #metoo world

Words by Stuart Heritage

Hello! If youre anything like me, youll be waking up in the middle of night worrying about climate change, plastic pollution and why the world is such a horrible place. Ive just written a book called Bedtime Stories for Worried Liberals, a collection of short funny stories about this. One of the stories is called The Man Who Couldnt Even Hug Anyone Any More, about a middle-aged white man (well, Im 39 now) struggling to adapt in a post #MeToo world. While writing, I realised that traditional masculinity is responsible for a huge slice of the worlds ills. So, heres my letter to my two young sons, about how Id like them to grow up

Dear sons,

Im sorry. This is all my fault. Had you been born at literally any other point in human history, you would have had the run of the place by now. As a pair of boys and relatively white boys at that the whole world has always been purpose built for your needs from the ground up.

Seriously, it was crazy what people like us used to get away with. We had all the money and all the power. If we wanted to go around groping women, we could. If we wanted to go out to work and leave our wives to stay at home and raise the kids, we could. Get this: if we wanted to own a new country, we used to just find one and tell everyone that it was ours. We did it all the time! And people actually let us! Isnt that nuts? This is the world you could have been born into.

But no. I met your mum too late, and you were born too late, and now thats all disappeared. We live in a post #MeToo world now, and Im afraid to say that theyre on to us. Youre the first generation of men in history who wont get to swan around doing whatever they like without fear of reprisal. I know. Its my fault. Im sorry.

This basically leaves you with two options. The first is to rail against your predicament, spluttering that equality is a sign of political correctness gone mad and that white men are actually now the real minority. Id advise against this, though, on the basis that itll make you look like a right tit. Your other choice, however, is to try and figure out how to be good, strong, considerate men in the world. Hopefully your mother and I have already shown you how to do this. But heres a reminder, just in case:

Being a man can suck sometimes. Our role models have always been strong and silent. Were taught to push our feelings all the way down to the pit of our stomach. Were told to man up. And this is devastating. If we dont talk about it, all our sadness and frustration at the world will have nowhere to go, and it ends up coming out in horrible ways. Sometimes it makes us hurt other people. Sometimes it makes us hurt ourselves. Youve seen me swear at strangers in the car before, so you can probably see Ive still got some work to do in this area. But I want you both to know that you can talk to me about your feelings. Im always going to be here for you.

You are both such beautiful, weird, tender little boys. Im proud of how thoughtful and sensitive you are, and I never want you stop being yourselves. But the day is bound to come when men will start bullying you to be more like them. Maybe theyll take against your all-consuming infatuation with bowhead whales. Maybe theyll knock the books out of your hands and tease you for not liking sport. This happened to me, and I ended up caving in to their demands; my entire school life was essentially spent pretending to understand football. Id love for you to be able to do better than me. You dont need to bow to the rigid demands of masculinity. You can like anything you want to like. You can wear whatever you want to wear. You can love whoever you want to love. Stick up for yourselves. Be better than me. And, if you cant be better than me, do what I do and use Facebook to see how badly all your old classmates have screwed up their lives. Its a lot of fun, I promise.

This is a big one. If I believe anything at all, its this. Instagram is full of people who think the best way to teach a three-year-old how to be decent is to photograph them holding a book about feminism that was written for 15-year-olds. Its infuriating. That isnt how children learn. They learn by watching and mimicking their role models. When you both grow up to be decent men, I hope that itll be in part because you saw me trying my hardest to be a good man. There are studies showing that children with engaged fathers have better cognitive development and more satisfactory relationships. Just by watching me cook dinner every night, for example, youre subconsciously learning not to believe in traditional gender roles. Youll grow up smart and self-reliant, and less likely to push the burden of emotional labour onto your partners. And your kids, if you have them, will see you doing it and theyll grow up to be even better than you. This is how change works. I guess what Im trying to say and I want you to read this slowly, so it really sinks in is that Im pretty bloody amazing

As I write this, youre both incredibly into superheroes. You call them brave heroes, which is actually quite adorable. But try to remember that superheroes arent brave. Batman isnt brave; hes a bored billionaire with nothing to live for. The Hulk isnt brave; hes just strong and stupid. Superman isnt brave; hes literally an invincible godhead from another planet. To truly be brave is to feel sad or scared, but find the strength to carry on anyway. Youre both already so brave: you were brave on your first day of nursery, and on your first day of school, and when you shouted down the bigger boys who pushed you at soft play, and when you saw that dead crab on the beach that time. If you can keep this spirit of bravery alive within you for your entire life then, my god, youll turn out to be great men.

I mean, youd think that this one is just common sense. But it bears repeating. Do not, under any circumstances, send a picture of your dick to a stranger on the internet. Its weird and gross. Final warning.

I love you both so much

Dad

PS. I swear to god, though, if youre still waking me up at four thirty every morning when you read this, Im cutting you out of my will.

Stus book Bedtime Stories For Worried Liberals, is out Thur 3 Oct, 7.19 (hardback) on Amazon normal RRP 9.99, published by Profile

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What Is the Root of Cronyism? – New Ideal

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If theres one issue that almost everyone agrees on whether Republican or Democrat, liberal or conservative its that cronyism is a significant problem in our political system, which is corrupted by special interest politics and rigged in favor of the well-connected and powerful.

But how well do we truly understand the nature and causes of cronyism, or how to solve the problem?

A thorough exploration of this question, and Ayn Rands distinctive perspective on it, can be found in the book Foundations of a Free Society: Reflections on Ayn Rands Political Philosophy, edited by Gregory Salmieri and Robert Mayhew. Cronyism is the focus of one essay in the collection, The Aristocracy of Pull: An Objectivist Analysis of Cronyism, by Steve Simpson, senior attorney at Pacific Legal Foundation and former director of Legal Studies at the Ayn Rand Institute.

Simpsons essay explains the origins of the corruption in our political system and points us toward the antidote.

As Simpson explains:

Ayn Rand viewed this issue radically differently from other thinkers and thus approached it in a fundamentally different way. She did not use the term cronyism and likely would not have used it, as it implies that the cause of problems such as pressure group warfare, influence peddling, and the unjust laws that result is individual favoritism. Instead, Rand looked for the cause of these problems in mistaken philosophical premises about the nature, purpose, and proper functions of government. In Rands view, the fundamental cause of these problems is not corrupt individuals but, rather, a flaw in the ends that government is held to serve. Increasingly, our laws and policies are based on and justified by altruism and collectivism, Rand argued. These premises lead to a political system that is designed to compel individuals to sacrifice their incomes, their labor, and ultimately their lives for the good of society. Rand saw any political system based on altruism and collectivism as a form of institutionalized thuggery a system in which some people possess the legal authority to impose their will by force on others. In any society, this will lead to a form of gang warfare, as different factions fight to control the government and thus the legal authority to sacrifice others.

Unlike the typical perspectives on cronyism we encounter today, Rands view is that the problem is deeply philosophical. Simpsons essay brings clarity to the issue, explains the origins of the corruption in our political system, and points us toward the antidote.

The Aristocracy of Pull is a must-read for anyone who wants to understand the root of and the solution to the deeply ingrained problem of cronyism. To learn more, buy your copy of Foundations of a Free Society today.

The author would like to acknowledge the useful editorial feedback of Keith Lockitch in improving this article.

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Marine Serre, Telfar, Draw Inspiration From the Apocalypse – Papermag

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Have you heard that the world is ending? It is! The climate strikers are running around asking for a merciful future for Leonardo DiCaprio's baby girlfriends; politicians are nightmares; it's fall and the weather in New York is set to hit 90 degrees on Wednesday. Hope is a dangerous thing for a woman like me to have and I don't have it! Rising seas, take me away.

Politics can be a dangerous thing for fashion designers to meddle with. Some do it adeptly; more make meaningless faux-woke slogan t-shirts produced by children in Bangladesh (and those are the ostensible liberals it's even funnier when conservative values creep in, like all those Ayn Rand t-shirts that they still sell at Brandy Melville). Fashion is an industry that pollutes the environment and exploits workers at all levels; enjoyment of it comes from the beauty, the fantasy, the artistry.

But fashion, as many have said before, doesn't exist in a bubble, and designers get inspired by current events, bleak though they may be. It's not universal as Sarah Kent and Zoe Suen wrote for Business of Fashion, the London shows generally seemed to take a break from "political navel-gazing," largely ignoring the incomprehensible mess that is Brexit. Milan shows were mostly about Italian glamour, minus the straitjackets and Foucault-quoting show notes at Gucci. But in New York and Paris, designers were getting ready for the end.

There were some sickos Bstroy, a "neo-native post-apocalypse streetwear brand," made headlines for making sweatshirts inspired by the worst elements of our current dystopia, i.e. school shootings (hoodies with Sandy Hook and Columbine logos were emblazoned with bullet holes). But most brands were far more thoughtful. While the Andreas Kronthaler for Vivienne Westwood Spring 2020 show in Paris seemed to mainly focus on the fantastical (the show notes described closer Bella Hadid as a "naked Fifties pin-up, photographed from below, looking like she's in the sky with puffs of cotton wool thrown on her"), the brand's recently released men's Spring 2020 lookbook stayed true to its history of protest and cries for sustainability; models were positioned underneath signs with slogans like "What's bad for the planet is bad for the economy."

For the past few seasons, Marine Serre has explicitly stated that her clothing is inspired by the impending end of the world as we know it. "It's after the apocalypse, a group of friends are underground a community coming together. It's a safe zone in which a new world is being created, a future world, a new way to see fashion," she said of her Fall 2019 collection.

For Spring 2020 (titled "Marre Noire," oil spill in English), the designer expanded on the theme, dressing survivors of the climate wars (including dogs) in her trademark "futurewear." The clothes, made from largely upcycled fabrics, were chaotic utilitarian the show began with black, militaristic garments, including a jumpsuit worn by an older, angry-looking (sexy) Frenchman who looked prepared to lead some sort of hacker revolution.

The Telfar spring 2020 show looked to migrants and immigration policies; models walked while a film played that featured TSA screenings and airport interrogations. And the opening looks were practical, clothes you could theoretically wear in the wake of global geopolitical collapse; designer Telfar Clemens showed four variations of cargo pants in a row. Cargo pants, which are both a callback to the Bush years and a garment that you could theoretically wear while on the run from fires and water wars, have been everywhere: brands ranging from Gypsy Sport to Maryam Nassir Zadeh (two downtown New York labels that have basically no aesthetic commonalities) have included them in the spring collections.

Utilitarian garments seem like an appropriate response to the climate catastrophe (they're also outdoorsy I guess we should all get outside and hike while we still can?). But they have another point of origin: lesbian style.

Sorry to generalize, but lesbians, as a group, represent some of the chicest trends of all. Lesbian aesthetics are vital: tailored suiting, Prada shoes, Vacancy Project haircuts. But ancient stereotypes of lesbian dressing could be seen in the spring collections, like the cargo pants and practical footwear (Tevas and Crocs have been trendy for a minute). You could see echoes of dyke camp.

In her popular 2018 essay "Notes on dyke camp," writer Mikaella Clements defined her theory of dyke camp, "a specific brand of humor, manners, and sensibilities guided by lesbian identity."

"Rather than drag, which parodies what is real dyke camp takes the real and magnifies it, so that it becomes absurd or funny or simply attractive in its own right," Clements wrote. "Dyke camp is a Walkman, a pair of Adidas sweatpants, the paintings of Kelly Beeman with their glazed sleepy stares, and Janelle Mone's PYNK video with its vagina pants. Big silver thumb rings are dyke camp, as are certain brands of gardening gloves; for obvious reasons, dyke camp tends to favor hands."

Designers may cite depressing forms of inspiration as opposed to queer aesthetics when it comes to their garments with zippers and pulleys and big pockets. But as Clements wrote, "the more you look for dyke camp, the more it seems to show up. Prowling, smirking, swaggering and usually already looking back at you."

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Marine Serre, Telfar, Draw Inspiration From the Apocalypse - Papermag

Joker is a film of our time, but not the film we need – Fabius Maximus website

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Summary: Joker is a film of our time. But it is an entertaining horror film feeding our fears (like drinking while depressed), not the inspirational film we need to defeat our fears. We have what we need on our shelves, waiting for us to use them.

The Batman stories evoke our fears for the future. Gotham City looms as a likely future for the America we are building, a high-tech society that abandoned its roots in Western values and so lost most of its social cohesion. All that remains are greed and power. The rich read Ayn Rand and feel superior while devoting themselves to conspicuous spending and collecting art. Our government devotes itself to gathering power over all things great and small, foreign and domestic. Inequality reaches Latin American levels that destroy the governments legitimacy. The underclass grows, becoming wilder. The shrinking middle class suffers impotently between those above and below.

In a world where science killed God and Nietzsche destroyed the Enlightenments lessons, we live in darkness above a void. Rational analysis no longer illuminates our lives. This is the story of the new film, Joker, about our existential fears given human form.

Now for the bad news: many Americans find the burdens of self-government too great to bear. Our new national motto seems to be Its not my fault. It should replace E Pluribus Unum on the dollar bill. This is the ethos of a nation in decline. It is why so many people fear for Americas future.

When a peoples conceits and delusions burn away, we fall back on our core beliefs: belief in freedom, free markets, human rights, and a republican form of government. But this aspect of America is an intellectual project. It has a strong hold on our minds but not on our hearts. That is not enough to break us out of our current decline. America is like a jet aircraft with sputtering engines, pilots bickering, and passengers panicking. Neither self-interest or love of our nation provides sufficient strength in such a crisis.

All we have left are myths. Unfortunately, our modern myths reflect the spiritual weakness that is one cause of our crisis. For example, see the stories of super-heroes that fill our theaters and TVs. Most of them tell about people who find a magic dingus and become great, or have powers bestowed on them by some Great MacGuffin. Heroes like Harry Potter, Shazzam, Spiderman, etc. James Bowman calls these Hollywoods slacker heroes.

In Xxx

These are entertainment, but not the kind that inspires or provides any guidance for our lives. It is not culture in its original meaning. Allan Boom explains this in his great work, Closing of the American Mind

{Culture is} everything that uplifts and edifies a people, as opposed to commerce. It constitutes a people, binding individuals into a group with roots, a community in which they think and become a moral unity of which the arts are an expression. It is the peak expression of mans creativity, our ability to break out of natures narrow bonds, and hence out of the degrading interpretation of man in modern natural and political science. It is profounder than the modern state, which deals only with mans bodily needs and tends to degenerate into mere economy.

American culture has myths that better match our past and can lead us to a greater future. We have myths that provide stronger food for our spirit and imagination. Here are two. You can list many more.

As a young boy, Bruce Wayne watched the murder of his parents. He resolved to prevent other children from suffering as he did, and spent years studying and training to become Batman. The story of a man voluntarily devoting his life to healing our broken society even by the most arduous and dangerous public service has great appeal. Its consistent with the admiration of Americans for the US military, who are despite their many failings the most trusted of our institutions.

James T. Kirk studied for years before entering Star Fleet Academy, preparing himself to become a great Starship Captain. As an instructor at the Academy, his students saw him as a stack of books with legs. He was familiar with both ancient philosophy (Spinoza, as mentioned in the TV episode Where No Man has Gone Before

There are other myths out there that can help, some from other lands. For example, we have a generation growing up many of whom saw the Fullmetal Alchemist TV shows and films (see Wikipedia), whose tagline (slightly paraphrased) is the kind of insight on which great nations can be built.

Humankind cannot gain anything without first giving something in return. To gain anything, something of equal value must be given. That is lifes First Law of Equivalent Exchange, and applies to things tangible and intangible matter, energy, and spirit.

These are just stories, but they represent a part of us to which we can look for inspiration in the dark times ahead.

People need stories, more than bread, itself. They teach us how to live, and why. Stories show us how to win. The Master Storyteller in HBOs wonderfulArabian Nights.

If you liked this post,like us on Facebookandfollow us on Twitter. See all postsabout heroes, aboutreforming America: steps to newpolitics, and especially these

By Joseph Campbell (1949).

This is the book that sparked serious research in to the function and significance of myths. See Wikipedia. From the publisher.

Since its release in 1949,The Hero with a Thousand Faceshas influenced millions of readers by combining the insights of modern psychology with Joseph Campbells revolutionary understanding of comparative mythology. In these pages, Campbell outlines the Heros Journey, a universal motif of adventure and transformation that runs through virtually all of the worlds mythic traditions. He also explores the Cosmogonic Cycle, the mythic pattern of world creation and destruction.

As relevant today as when it was first published,The Hero with a Thousand Facescontinues to find new audiences.

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Joker is a film of our time, but not the film we need - Fabius Maximus website

SUNY Potsdam chemistry professor awarded grant from National Institutes of Health – NNY360

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POTSDAM Dr. Fadi Bou-Abdallah, a professor in the Department of Chemistry at SUNY Potsdam, has received a grant award from the National Institutes of Health for $414,047.

The NIH Academic Research Enhancement Award grant will fund Bou-Abdallahs project, titled Effect of Ferritin Subunit Composition on Iron Core Formation, Morphology and Iron Mobilization: Physical Characterization and Physiological Relevance, for the next three years (from 2019 to 2022).

Through this research, Dr. Bou-Abdallah seeks to understand how ferritin, the major iron storage protein in mammals, plays a crucial role in iron mineralization and housekeeping.

Improper iron mineralization has been suggested to contribute to the progression of diseases, and a number of studies have found abnormal brain iron and ferritin levels in neurodegenerative disorders, such as Parkinsons, Alzheimers disease and Friedreichs ataxia, Bou-Abdallah said.

These abnormalities have been discussed in terms of a loss of ferritins ability to maintain iron homeostasis, leading to ferritin and iron deposits in brain tissues. The proposed research will help to define the molecular origins of these debilitating diseases, and provide fundamental insights into the biochemical processes responsible for iron-related disorders and neurodegenerative diseases, Bou-Abdallah said. The results would also facilitate exploitation of ferritin as a nanotemplate, for uses in nanochemistry, nanobiology and nanomedicine.

It is a great honor to be recognized by NIH for the wonderful science we do at SUNY Potsdam with our undergraduates. This grant award would not have been possible without our students dedication and hard work, and represents a culmination of years of excellence in undergraduate teaching and research, Bou-Abdallah said. Support of undergraduate research at small institutions like ours is extremely important. The majority of students who receive bachelors degrees in our department go on to earn doctorates and/or medical degrees. Undergraduate research is a fundamental part of their education at SUNY Potsdam, and one of the high-impact learning practices they encounter here.

Bou-Abdallah plans to work with SUNY Potsdam undergraduates over the next three academic years and summers to complete the project. The students will be involved in all aspects of this research and be exposed to interdisciplinary research at the interface of chemistry, biology and materials science. They will also get a chance to travel to the University of Pennsylvania and work with experts in the fields of biosciences and materials science, and use a powerful scanning transmission electron microscope that provides images down to the atomic level, the first of its kind in the United States.

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SUNY Potsdam chemistry professor awarded grant from National Institutes of Health - NNY360

Improving 131I Radioiodine Therapy By Hybrid Polymer-Grafted Gold Nano | IJN – Dove Medical Press

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Marine Le Goas,1 Marie Paquet,25 Aurlie Paquirissamy,1 Julien Guglielmi,24 Cathy Compin,24 Juliette Thariat,6 Georges Vassaux,24 Valrie Geertsen,1 Olivier Humbert,25 Jean-Philippe Renault,1 Graldine Carrot,1 Thierry Pourcher,24 Batrice Cambien24

1NIMBE, Commissariat lEnergie Atomique, Centre National Recherche Scientifique UMR 3685, Universit Paris-Saclay, Gif-sur-Yvette, France; 2Laboratory Transporter in Imaging and Radiotherapy in Oncology (TIRO), Institut de Biosciences et Biotechnologies dAix-Marseille (BIAM), Commissariat lEnergie Atomique, Nice, France; 3Laboratory Transporter in Imaging and Radiotherapy in Oncology (TIRO), University Nice Sophia Antipolis, Nice, France; 4Laboratory Transporter in Imaging and Radiotherapy in Oncology (TIRO), University Cte dAzur, Nice, France; 5Nuclear Medicine Department, Centre Antoine Lacassagne, Nice, France; 6Department of Radiation Oncology, Centre Franois Baclesse, Universit de Normandie, Caen, France

Correspondence: Batrice CambienLaboratory Transporter in Imaging and Radiotherapy in Oncology (TIRO), University Nice Sophia Antipolis, 28 Avenue Valombrose, Nice Cedex 2 06107, FranceTel +33 493 377 715Email cambien@unice.fr

Background: Human trials combining external radiotherapy (RT) and metallic nanoparticles are currently underway in cancer patients. For internal RT, in which a radioisotope such as radioiodine is systemically administered into patients, there is also a need for enhancing treatment efficacy, decreasing radiation-induced side effects and overcoming radio-resistance. However, if strategies vectorising radioiodine through nanocarriers have been documented, sensitizing the neoplasm through the use of nanotherapeutics easily translatable to the clinic in combination with the standard systemic radioiodine treatment has not been assessed yet.Method and materials: The present study explored the potential of hybrid poly(methacrylic acid)-grafted gold nanoparticles to improve the performances of systemic 131I-mediated RT on cancer cells and in tumor-bearing mice. Such nanoparticles were chosen based on their ability previously described by our group to safely withstand irradiation doses while exhibiting good biocompatibility and enhanced cellular uptake.Results: In vitro clonogenic assays performed on melanoma and colorectal cancer cells showed that poly(methacrylic acid)-grafted gold nanoparticles (PMAA-AuNPs) could efficiently lead to a marked tumor cell mortality when combined to a low activity of radioiodine, which alone appeared to be essentially ineffective on tumor cells. In vivo, tumor enrichment with PMAA-AuNPs significantly enhanced the killing potential of a systemic radioiodine treatment.Conclusion: This is the first report of a simple and reliable nanomedicine-based approach to reduce the dose of radioiodine required to reach curability. In addition, these results open up novel perspectives for using high-Z metallic NPs in additional molecular radiation therapy demonstrating heterogeneous dose distributions.

Keywords: internal radioisotope therapy, radioiodine, polymer-grafted gold nanoparticles, melanoma, colorectal cancer, radio-enhancement

This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License.By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms.

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MagForce AG to host Lunch Symposium on Local Therapies in Malignant Gliomas during the 19th European Congress of Neurosurgery (EANS2019) – BioSpace

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Berlin, Germany and Nevada, USA, September 19, 2019 - MagForce AG (Frankfurt, Scale, XETRA: MF6, ISIN: DE000A0HGQF5), a leading medical device company in the field of nanomedicine focused on oncology, is pleased to announce that it will host a scientific lunch symposium titled "Local Therapies in Malignant Gliomas - update and new perspectives" during the 19th European Congress of Neurosurgery in Dublin, Ireland.

Chaired by Prof. Dr. Walter Stummer, Director of the Department of Neurosurgery at the University Hospital of Mnster (UKM), Germany, the one-hour lunch symposium will feature two keynote speeches: After an introduction and overview on the current status of glioma treatments by Prof. Dr. Stummer, Ricardo Dez Valle, MD PHD, Head of the Department of Neurosurgery, Hospital Group Quirn Madrid, Spain, will give an update on local neurosurgical therapies. The symposium will be rounded off with a talk from Prof. Dr. Stummer on An Emerging Adjunct: NanoTherm NanoPaste Application. During the subsequent wrap-up and discussion, participants will have the opportunity to ask questions.

Symposium details:

Title:

Local Therapies in Malignant GliomasUpdate and new perspectivesNanoTherm - NanoPaste Application

Date:

Wednesday, September 25, 2019

Time:

1.15-2.15 p.m. local time/GMT

Location:

EcoCem meeting room, The Convention Centre Dublin

Speakers:

Prof. Dr. Walter Stummer (Chair)University Hospital of Mnster, GermanyIntroductionCurrent Status of Glioma Treatments

Ricardo Dez Valle, MD PHDHospital Group Quirn Madrid, SpainLocal Neurosurgical Therapies: An Update

Prof. Dr. Walter StummerUniversity Hospital of Mnster, GermanyAn Emerging Adjunct: NanoTherm NanoPaste Application

In addition to the lunch symposium, MagForce will host Meet the Expert sessions on Thursday, September 26th and Friday, September 27th, at 10 a.m. respectively, at the MagForce booth No. 35 located at Level 0 The Forum. On both days, Dr. Michael Schwake, Senior Physician in the Department of Neurosurgery at the University Hospital of Mnster and member of Prof. Dr. Stummers team, will be available to interested participants for questions as an expert in the use of the NanoTherm therapy system in clinical practice.

Prof. Dr. Stummer and his team at the University Hospital of Mnster have been treating brain tumor patients with MagForce's NanoTherm therapy since early 2015 and were the first to introduce, clinically, the new nanoparticles application technique called 'NanoPaste'. In previous clinical research, the UKM team demonstrated that a better applicability of heat-focussing nanoparticles around the resection rim after surgical removal of a brain tumor could boost the thermotherapy treatment for tumor ablation and as a radiosensitiser in glioma therapy. With the use of the new NanoPaste technique, the team was able to create sufficient nanoparticle concentrations to reach effective thermotherapy in the glioma resection cavity wall and its vicinity, allowing for more control of remaining tumor infiltration. Especially in comparison to stereotactic techniques with their associated imponderabilities in scarred and pretreated tissues, the 'NanoPaste' application was technically easy, controllable and quick to perform.

About NanoTherm TherapyMagForce's NanoTherm therapy provides a novel, nanotechnology-based approach for the treatment of solid tumors by introducing magnetic nanoparticles either directly into the tumor or into the resection cavity wall. These particles are subsequently heated by an alternating magnetic field that allows targeted treatment by irreparably destroying, or weakening, cancer cells making them more sensitive to concomitant radiotherapy or chemotherapy. Since the particles remain at the site of application due to their special coating, the surrounding healthy tissue is spared and repeated treatments as well as the integration into multimodal therapy concepts are made possible. MagForce AG was granted the European CE certificate ("European Certification") in 2011 and thus the official approval of NanoTherm therapy for the treatment of brain tumors in Germany and all member states of the European Union.

About EANS2019The European Association of Neurosurgical Societies (EANS) is both an independent federation of European national neurosurgical societies and a fast-developing association of individual neurosurgeons from around the world, aiming to enhance the quality of neurosurgical patient care through training, education and research. A key way in which the association fulfils this objective is by facilitating the exchange of scientific information at the highest level through the organisation of meetings, symposia and educational courses with the annual congress as its flagship event.

The overall scientific programme will take the familiar form that has been established over the last few years bringing together international experts to discuss and inform. To help develop best practice in all subspecialities under the headings of Technology, Techniques, Training and increasingly important Transparency. The four days of scientific discussions offer plenty of opportunities for meetings, socialising and hearing of the latest scientific developments.

For more information on the EANS2018 programme, please click here: https://eans2019.com/programme-and-abstracts/scientific-programme.html.

About MagForce AG and MagForce USA, Inc.MagForce AG, listed in the Scale segment of the Frankfurt Stock Exchange (MF6, ISIN: DE000A0HGQF5), together with its subsidiary MagForce USA, Inc. is a leading medical device company in the field of nanomedicine focused on oncology. The Group's proprietary NanoTherm therapy enables the targeted treatment of solid tumors through the intratumoral generation of heat via activation of superparamagnetic nanoparticles.

NanoTherm, NanoPlan, and NanoActivator are components of the therapy and have received EU-wide regulatory approval as medical devices for the treatment of brain tumors. MagForce, NanoTherm, NanoPlan, and NanoActivator are trademarks of MagForce AG in selected countries.

For more information, please visit: http://www.magforce.comGet to know our Technology: video (You Tube)Stay informed and subscribe to our mailing list

DisclaimerThis release may contain forward-looking statements and information which may be identified by formulations using terms such as "expects", "aims", "anticipates", "intends", "plans", "believes", "seeks", "estimates" or "will". Such forward-looking statements are based on our current expectations and certain assumptions, which may be subject to a variety of risks and uncertainties. The results actually achieved by MagForce AG may substantially differ from these forward-looking statements. MagForce AG assumes no obligation to update these forward-looking statements or to correct them in case of developments, which differ from those, anticipated.

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MagForce AG to host Lunch Symposium on Local Therapies in Malignant Gliomas during the 19th European Congress of Neurosurgery (EANS2019) - BioSpace

Synthetic networks with tunable responsiveness, biodegradation, and molecular recognition for precision medicine applications – Science Advances

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INTRODUCTION

In 2015, the Obama administration launched the precision medicine initiative (1). An emerging engineering challenge within precision medicine is the need for versatile platform technologies that can be tailored to individual patients or pathologies (1, 2). A common approach within the fields of biomaterials and nanotechnology has been to design highly tailored formulations that target specific cell and tissue characteristics of a single pathology. These formulations, which can be fabricated in a variety of supramolecular structures [i.e., linear polymer conjugates (3), gels (4, 5), and self-assembled materials (6, 7)], recognize hallmark overexpressed cellular markers for the purpose of disease targeting. Nanoparticle carriers for precision medicine applications are typically dynamic in nature, swelling and/or degrading in intracellular environments to deliver therapeutic payloads to the cytosol of target cells (8).

In cancer treatment, there is precedence that multiple therapeutics can act synergistically to target and kill tumors. Chemotherapeutic agents act through a variety of mechanisms, including, but not limited to, DNA intercalation, enzyme inhibition, and cell cycle arrest (9, 10). Targeted agents, such as monoclonal antibodies, alter cell signaling pathways and engage the immune system. Photothermal therapy leads to tumor reduction by increasing membrane fluidity (~43C) or ablating the cells (~50C) (11). All of these therapeutic modalities benefit from targeting strategies, which concentrate the therapeutic agent within the tumor.

Each therapeutic option has distinct potential to aid in an individual patients treatment regimen. But, there is also marked variability between patients, necessitating precise and tailored treatments specific for the genetic and biophysical properties of the individual pathology. Advancements in genomic and proteomic technologies have made the collection of these relevant individual data a reality. The major hurdles left to overcome include, first, establishing predictive models of patients response to treatment and, second, engineering highly tunable platform technologies that deliver multiple therapeutic modalities in a patient-specific manner. Our modular strategy addresses the latter challenge and could serve as a useful tool in future studies on the former.

Previous studies on nanoparticle development for cancer precision medicine have focused on highly specified platforms that efficiently target and kill a single tumor population. For example, Conde et al. (12) recently designed a composite platform composed of gold nanorods, gold nanoparticles, therapeutic antibodies, and small interfering RNA encapsulated within an adhesive hydrogel patch. This system targeted and killed colorectal cancer cells through multiple modalities (i.e., photothermal therapy, RNA interference, and targeted chemotherapy), increasing treatment efficacy in vitro and in vivo.

In another illustrative example, Liu et al. (13) constructed a hierarchical nanomaterial assembly that delivered a cytotoxic protein (ribonuclease A) and antibiotic (doxycycline). This platform targeted cancer stem cells within heterogeneous cancer populations. The targeted, dual therapy led to a significant reduction in tumor volume relative to both the controls and individually administered therapeutics. These are only two examples, out of many promising studies on cancer nanomedicines that have used multiple therapeutic modalities (1215). There is a need, therefore, for a readily modifiable platform that facilitates the rapid customization of cancer nanomedicines to individual patients pathologies.

We previously demonstrated the ability to tune the hydrodynamic diameter and magnitude of pH response of poly(acrylamide-co-methacrylic acid) [P(AAm-co-MAA)] nanogels by modulating the monomer feed, polymerization parameters, or purification strategy (16). In the present work, our base platform is this random P(AAm-co-MAA) copolymer cross-linked into a nanogel with either a nondegradable or a redox-labile cross-linker. We present a new, modular sequence of nanogel modifications with small molecules, peptides, or proteins; these nanogels are multifunctional and multiresponsive, exhibiting dynamic loading and release of therapeutic payloads, engaging in a bioactive manner with biological substrates, transducing external signals into therapeutic heating, and promoting cellular internalization.

To achieve modular functionalization while retaining the bioactivity of conjugated molecules, we rely on facile and biocompatible conjugation schemes. While there are numerous bioconjugation strategies documented in the literature (1719), we use carbodiimide-mediated coupling to attach diverse ligands to pendant carboxylic acid groups via a stable amide bond. In this coupling scheme, carboxylic acid groups are activated with a catalyst to form a reactive ester intermediate, which is highly reactive with primary amines in slightly acidic aqueous solutions. We can, therefore, couple any water-soluble, amine-terminated moleculeincluding, but not limited to, proteins, peptides, and small moleculesdirectly to our polymer backbone. The diversity of bifunctional linker molecules that are available commercially, such as poly(ethylene glycol) derivatives (20, 21), further diversifies the ligands that our platform can accommodate, including those with amine, carboxylic acid, hydroxyl, or sulfhydryl groups.

We hypothesized that a single platform, when modified in a modular manner with bioactive components, could respond dynamically to tumor physiological environments, partition and elute therapeutic agents in a controlled manner, transduce external signals for therapeutic heating, and target tumor populations. We believe that this platformwhich can be modified to achieve environmental responsiveness, therapeutic delivery, and molecular recognitionis an enabling technology for delivering personalized and calibrated combination therapies. A summary schematic for our platform, along with the reagents, chemical modifications, and therapeutic modalities explored, is given in Fig. 1. In this proof-of-concept study, we demonstrate how a single, biocompatible platform can be quickly and precisely modified for personalized and precision medicine applications. Furthermore, in addition to standard characterization techniques, we developed and applied two new experimental methods: a quartz crystal microbalance with dissipation (QCM-D) assay for nanogel degradation and a high-throughput cell imaging assay for determining the kinetics of nanogel uptake. These new techniques expand upon the fields repertoire of experimental methods for evaluating and comparing new nanoparticle systems for precision medicine.

Nanoscale networks of acrylamide (AAm) and methacrylic acid (MAA), cross-linked with methylenebisacrylamide (BIS) or its degradable disulfide analog [N,N-bis(acryloyl)cystamine], were synthesized by inverse emulsion polymerization and modified via carbodiimide chemistry with tyramine (Tyr), N,N-dimethylethylenediamine (DMED), proteins, or peptides. In an additional post-synthesis step, gold nanoparticles (AuNP) were precipitated within DMED-modified (DMOD) nanogels. Here, we document the synthesis and modification of this nanogel platform and demonstrate the impact of nanogels modification on their ability to respond to the pH environment, load and release a model cationic drug, target cells, act as a functional enzyme, and transduce green light for photothermal therapy. Because of its tunability and the variety of therapeutic modalities enabled, we believe that this platform is suitable for precision medicine applications. DTT, dithiothreitol; TMB, 3,3,5,5-tetramethylbenzidine.

Our base platform for small moleculemodified nanogels was an ionomer collapsepurified P(AAm-co-MAA) nanogel, synthesized as described by Zhong et al. (16). These nanogels had a swollen hydrodynamic diameter of 768 nm, were 63% acidic copolymer by mass, and exhibited pH-responsive expansion/syneresis behavior with a critical pH transition point of 4.8. Ionomer collapsepurified nanogels, as opposed to those purified by dialysis alone, were selected because the basic conditions (0.5 N sodium hydroxide) are known to induce hydrolysis of some of the nanogel acrylamide content to acrylic acid, providing additional acid groups for bioconjugation. The increased presence of carboxylic acids allowed us to couple a greater quantity of functional small molecules to each nanogel.

An additional degree of tunability was introduced to the P(AAm-co-MAA) nanogels by introducing a biodegradable cross-linker. N,N-bis(acryloyl)cystamine is a bisacrylamide analog that contains a labile disulfide linkage. It has been used previously as a component of digestible gels for drug and gene delivery applications (2224). We successfully cross-linked P(AAm-co-MAA) nanogels with N,N-bis(acryloyl)cystamine. These biodegradable nanogels were similar in hydrodynamic diameter, zeta potential, and pH-responsiveness to their nondegradable analogs (fig. S1).

The kinetics and mechanisms of biodegradation for these nanogels were quantified by optical and gravimetric analyses. Optical analysis was conducted via dynamic light scattering (DLS) with a fixed detector position and signal attenuation. Under these measurement conditions, the count rate is related to the decrease in the number of suspended nanoparticles (25). Simultaneously, the hydrodynamic diameter measurements collected provide inference to the mechanism of biodegradation (i.e., surface erosion and bulk degradation). We assessed biodegradation by DLS for degradable nanogels in the presence of 10 mM dithiothreitol (DTT) or glutathione in 1 phosphate-buffered saline (PBS) at pH 7.4. DTT is a common reducing agent used for biological applications. It has been used previously to degrade systems cross-linked with N,N-bis(acryloyl)cystamine and was selected to ensure that the nanogels degrade completely. Glutathione (10 mM) in 1 PBS was selected as a biologically relevant reducing condition, as it mimics the intracellular environment (26).

The chemical mechanism of nanogel degradation by each reducing agent, as well as a pictorial depiction of the biodegradation process, is given in Fig. 2A. DLS analysis confirmed that both DTT and glutathione were able to reduce the disulfide cross-linker and consequently degrade the nanogel network (Fig. 2B). In the presence of DTT, the nanogels degraded rapidly and were indistinguishable from a linear polymer solution of the same concentration (i.e., completely degraded) after 40 min. Nanogels degraded with reduced kinetics in a 10 mM glutathione solution. The normalized count rate declined by 72.05.8% after 50 min in glutathione solution, and the nanogels were indistinguishable (by DLS count rate) from linear polymer after 48 hours.

(A) N,N-bis(acryloyl)cystamine cross-linked nanogels degrade via reduction of the disulfide. The diagram demonstrates how, after an initial period of surface erosion, the nanogels experience bulk degradation, leading to simultaneous network swelling. (B) DLS analysis of nanogel degradation. While bisacrylamide cross-linked nanogels did not degrade under reducing conditions, those cross-linked with a disulfide cross-linker were digested by both reducing agents (n = 4, mean SD). (C) QCM analysis demonstrated the kinetic decomposition of nanogels under reducing conditions and flow. While the mass of nondegradable nanogels was relatively unaffected by reducing conditions, the mass of degradable gels declined rapidly (n = 3, mean SD). (D) Hydrodynamic diameter analysis by DLS supported the degradation mechanism of initial surface erosion followed by bulk degradation. While the normalized count rate declined steadily throughout the extended measurement, the hydrodynamic diameter decreased initially (surface erosion) and then increased for the remainder of the experiment (i.e., decrease in cross-links led to a reduction in the total number of nanoparticles but swelling of the remaining intact nanogels) (n = 3, mean SD).

In QCM-D experiments, the nanogels were covalently conjugated to a gold-coated quartz sensor, and the mass loss, under reducing conditions, was monitored by measuring the change in the quartz sensors fifth harmonic resonance frequency (27). Mass loss was normalized to the initial mass of coupled nanogels to determine a relative measure. It is noteworthy that because the nanogels are covalently conjugated to the quartz sensor, the mass loss will never reach 100%. Some linear polymer strands will remain immobilized on the sensor following complete degradation of the cross-links.

Under a steady flow of fresh 10 mM DTT (1 PBS, pH 7.4), the mass of nondegradable nanogels increased slightly. This increase was likely due to adsorption of DTT molecules. On the other hand, the relative mass of degradable nanogels decreased rapidly, reaching a degraded state in 15 min (Fig. 2C). As shown in fig. S2, in parallel with an increasing resonance frequency, the dissipation of sensors coated with degrading increased. This indicated that as the nanogels were degrading, they were simultaneously losing mass and imbibing water. This observation was consistent with our DLS measurements, which showed that the nanogels simultaneously degraded and swelled under reducing conditions (Fig. 2D).

Next, we explored the ability to modify the pendant acid groups on P(AAm-co-MAA) with amine-terminated small molecules (tyramine and N,N-dimethylethylenediamine) to add phenol or tertiary amine groups to the polymer backbone, respectively. Nanogels modified to different extents with tyramine (TMOD) or N,N-dimethylethylenediamine (DMOD) were characterized by Fourier transform infrared (FTIR spectroscopy), potentiometric titration, DLS, and zeta potential measurement (Fig. 3).

(A) FTIR spectra of TMOD and DMOD nanogels, as compared with the unmodified formulation. The peaks at 1700 and 1200 cm1 correspond to the carboxylic acid, at 1660 and 1590 cm1 correspond to the amide, and at 800 cm1 correspond to the aromatic groups, confirming the incorporation of each small-molecule ligand through covalent coupling. FTIR analysis of all formulations is presented in fig. S1. (B) Nanogel modification proceeded with approximately 60% efficiency when the ligand concentration did not exceed the carboxylic acid concentration (stoichiometric ratios less than 1). (C) Potentiometric titrations were used to quantify the carboxylic acid content of all formulations, elucidating the extent of small-molecule coupling. (D) Modified nanogels exhibited a pH-responsive zeta potential transition (anionic to cationic), whereas unmodified nanogels were anionic across all pH values tested (n = 3, mean SD). (E) Unmodified and TMOD nanogels exhibited a pH-responsive collapse with a critical transition point at pH ~ 4.8. DMOD nanogels did not undergo substantial pH-responsive swelling.

FTIR analysis showed that the small molecules were covalently bound to the nanogel network, as evidenced by the reduction in peaks corresponding with the carboxyl carbonyl (1700 cm1) and carbon-oxygen single bond (1200 cm1). A graphical depiction of select formulations is given in Fig. 3A, with the full analysis of all formulations given in fig. S3. From the FTIR analysis, it initially appeared that the DMOD reaction proceeded with greater efficiency, as the disappearance of carboxylic acid peaks was more pronounced. However, potentiometric titration revealed that the percent of modified methacrylic acid moieties trended with the stoichiometric ratio of ligand to carboxylic acid similarly for both ligands (Fig. 3, B and C). Therefore, the trends observed in the FTIR spectra are likely reflective of the location of nanogel modification (surface for DMOD and bulk for TMOD) rather than the efficiency of the reaction. The circled formulations (0.5 TMOD and 0.78 DMOD) in Fig. 3B, which achieved a high degree of efficient molecular coupling, were used in each of the following experiments.

Potentiometric titration, pH-responsive zeta potential, and size measurements for TMOD, DMOD, and unmodified nanogels are also presented in Fig. 3 (C to E). As shown in the potentiometric titration analysis, unmodified nanogels were 63% polyacid [i.e., poly(acrylic acid) and poly(methacrylic acid)] by mass, as compared with 22 and 25% for the TMOD and DMOD nanogels, respectively. The reduction in acid content, because of modification, trended linearly with ligand concentration in the modification reaction at low extents of modification and plateaued at 69.83.7% modification. Full potentiometric titration analysis of all TMOD and DMOD formulations is presented in fig. S4.

All three formulations were anionic at pH values greater than five, as the carboxylic acid groups were predominantly deprotonated [pKa (where Ka is the acid dissociation constant), ~4.8] and held a negative charge. As the pH was reduced from 8 to 3, the TMOD and DMOD nanogels zeta potential was less negative than that of unmodified nanogels and became positive at pH 4.7. In this pH environment, as well as those more acidic, the carboxylic acid groups are protonated and therefore uncharged, whereas the tertiary amines contributed by N,N-dimethylethylenediamine and adsorbed sodium ions from the solution (5 mM sodium phosphate buffer) are positively charged. This pH-responsive ionization change for the modified nanogels is especially critical for environmentally responsive drug delivery, as will be shown in a later section.

The modified and unmodified nanogels hydrodynamic diameters, as measured by DLS, also changed in response to the pH environment. TMOD and unmodified nanogels exhibited similar pH-responsive collapses, with a critical pH transition of approximately 4.8. As the pH of the solution was dropped below 4, both the TMOD and DMOD nanogels aggregated. For the purpose of visualization, hydrodynamic diameter measurements from aggregated states were omitted from Fig. 3E. The full data are presented in fig. S5.

It is noteworthy that DMOD nanogels exhibited a nearly complete loss of pH-responsive swelling. This can be attributed to the fact that because of the modified networks amphoteric nature, it bears charge across all pH values. Its state of electrical neutrality at pH 4.7 is a result of balanced negatively and positively charged species, rather than a loss of ionization. On the contrary, the pH-responsive behavior of both the unmodified and TMOD nanogels suggests aggregation caused by a hydrophobic transition and loss of ionization. Taken in combination with the observed trends in zeta potential, this suggests that the TMOD nanogels negative-to-positive charge transition is a result of the association of ionic species, both salts from the buffer and additional tyramine molecules that were neither conjugated nor extracted during purification, rather than the network components themselves bearing a positive charge.

Suspensions of DMOD, TMOD, and unmodified nanogels were incubated separately with methylene blue in distilled water, and methylene blue loading was achieved through equilibrium partitioning. Methylene blue was selected as a model therapeutic agent because of its cationic nature, use as a photosensitizer, and similarity to the chemotherapeutic 5-fluorouracil. Methylene blue is a hydrophilic compound (logP=1.1), similar to 5-fluoruracil (logP=0.89). Methylene blueloaded nanogels were dialyzed against 1 PBS (of pH 4.5 or 7.4), which was exchanged regularly with fresh buffer to both simulate drug sequestration/metabolism and establish a semi-sink condition. The buffer condition (1 PBS at pH 7.4) was intended to simulate the pH environment in circulation, whereas the pH 4.5 condition was meant to emulate the environment of the late endosome, which nanocarriers will experience during lysosomal trafficking following cellular uptake. It is noteworthy that in the case of cancer drug delivery, the nanocarriers will experience a gradient of pH, decreasing from circulation through the endosomal pathway. The drug release environment was maintained at 37C, and methylene blue elution was monitored until complete release was achieved (28 hours).

Unmodified nanogels loaded significantly more methylene blue than their TMOD and DMOD derivatives (fig. S6). Prior to modifications, nanogels loaded methylene blue with 99.50.3% efficiency (equal mass ratio nanogels: methylene blue in ultrapure water). Increasing nanogel modification with tyramine or N,N-dimethylethylenediamine decreased the equilibrium partitioning of methylene blue. Specifically, TMOD and DMOD nanogels loaded methylene blue with 59.72.1% and 34.9 9.2% efficiency, respectively. This decrease in equilibrium partitioning, relative to unmodified control nanogels, is due to the hydrophobicity or cationic character that the respective ligands contribute. As a cationic and hydrophilic payload, methylene blue enages in complementary electrostatic interactions with deprotonated methacrylic acid groups. Furthermore, as methylene blue partitions preferentially in water over organic phases, we expected loading efficiency to correlate positively with nanogel hydrophilicity. In the preceding section, we showed that the extent of nanogel functionalization correlated with the amount tyramine or N,N-dimethylethylenediamine in the reaction solution. Furthermore, as each modification reaction depleted a pendant methacrylic acid group, there is a negative relationship between extent of nanogel modification and the available methacrylic acid groups to interact with methylene blue. Following modification with N,N-dimethylethylenediamine, the amphoteric nanogels lost pH-responsive swelling behavior. Their tertiary amine moieties, which are cationic, exerted a repulsive force on methylene blue. As a result of tyramine modification, the nanogels became more hydrophobic, similarly lowering the networks ability to partition methylene blue. These physicochemical characteristics of TMOD and DMOD nanogels are useful for responsive release behavior, but as they decrease the nanogelmethylene blue affinity, they decrease methylene blue loading efficiency.

A drug release experiment probed the ability of each modified or unmodified nanogel system to act as an intelligent drug delivery vehicle. In this experiment, nanogels loaded with methylene blue [nanogels (1 mg/ml), with corresponding loading described above] were placed in dialysis tubing [regenerated cellulose, molecular weight cutoff (MWCO), 12,000 to 14,000 kDa] and dialyzed against 1 PBS (pH 4.5 or 7.4). The dialysate was exchanged for fresh buffer every 2 hours to simulate drug metabolism. At each time point, a sample was taken from both within the dialysis tubing and outside it (i.e., the dialysate) to ensure precise measurement of the kinetic methylene blue release.

Unmodified nanogels exhibited sustained-release kinetics without a noticeable burst release or pH-responsiveness. In 2 hours, unmodified nanogels eluted 41.015.5% and 46.04.0% of their loaded methylene blue at pH 7.4 and 4.5, respectively (Fig. 4A). The unmodified nanogels consistency, in their rate of methylene blue elution between the two pH environments, is consistent with their continuously anionic zeta potential. While the pH 4.5 environment is below the unmodified nanogels pKa, sufficient acid moieties remained deprotonated to engage in electrostatic interactions with methylene blue and promote payload retention in a manner similar to the pH 7.4 condition.

(A) Methylene blue experienced complementary electrostatic interactions with unmodified nanogels at both pH 4.5 and 7.4, leading to sustained release in both conditions. (B) TMOD nanogels exhibited an initial burst release of methylene blue, where the quantity of that release was greater in acidic than neutral conditions. (C) DMOD nanogels exhibited a burst release of greater than 50% the loaded payload in each pH condition, with more rapid release in acidic than neutral conditions. (D) DMOD and TMOD nanogels exhibited similar methylene blue release behavior in acidic conditions, while unmodified gels exhibited a more sustained-release profile. (E) DMOD nanogels released methylene blue rapidly in 1 PBS (pH 7.4), while unmodified nanogels exhibited sustained-release and TMOD gels displayed intermediate behavior. The results in (D) and (E) indicated that the nanogels zeta potential is largely predictive for their release profile [all panels: n = 4, mean SD; *P < 0.05, **P < 0.01, and ***P < 0.001, two-way analysis of variance (ANOVA) with Tukey posttest].

On the other hand, TMOD nanogels exhibited an initial burst release, which varied significantly with the pH environment (78.59.6% and 43.7 18.9% of the loaded payload in the first 15 min, at pH 4.5 and 7.4, respectively; P < 0.05). After the burst release, TMOD nanogels gradually released methylene blue at pH 7.4 and rapidly released it at pH 4.5 (Fig. 4B). TMOD nanogels bear a cationic zeta potential in acidic buffers and further undergo a hydrophile-to-hydrophobe transition around the critical pH point (pH 4.8). These physical and chemical alterations, which were unique to the TMOD nanogels, explain their significant and unique pH-responsive methylene blue release profile. DMOD nanogels exhibited substantial burst release, followed by rapid methylene blue elution at both pH 4.5 and 7.4 (Fig. 4C). Methylene blue release was more rapid from DMOD nanogels at pH 4.5 than pH 7.4, which can be attributed to the transition from anionic to cationic zeta potential, as was shown previously.

Figure 4 (D and E) highlights the differences in pH-responsive methylene blue elution for the three formulations. All nanogel formulations eluted the entirety of the methylene blue payload within 28 hours. Unmodified nanogels exhibited a sustained-release profile in both pH conditions, demonstrating their use for controlled release but lack of responsive release. DMOD nanogels, conversely, released methylene blue rapidly in both pH environments, acting as neither a sustained-release depot nor a responsive delivery vehicle. TMOD nanogels acted as a pH-responsive delivery vehicle, responding to the acidic environment by rapidly releasing methylene blue. In the pH 4.5 environment, there were significant differences (P<0.001) between the relative elution of methylene blue from modified and unmodified nanogels. However, there were no differences between the methylene blue elution profiles of the DMOD and TMOD nanogels. This indicated that the release profile is driven primarily by the nanogels cationic zeta potential and not a hydrophile-to-hydrophobe transition (which was unique to TMOD). In the pH 7.4 environment, there were significant differences between the methylene blue elution profile of all three formulations.

These results illustrated how modification of the acid moiety, through changing the nanogels environmentally responsive swelling and ionization, altered the systems use as a drug delivery vehicle. While unmodified nanogels were most advantageous for steadily delivering a hydrophilic, cationic payload to the surrounding environment, TMOD nanogels exhibited rapid pH-responsive delivery. This pH responsiveness could lead to triggered release in the acidic tumor or endosome microenvironments. Therefore, the identity and extent of nanogel surface modifications should be carefully tuned to yield combinations of sustained and responsive release for specific drug delivery applications.

Our original design goal was to construct a tunable nanoscale hydrogel platform that was cytocompatible and could be diversified in a modular manner with bioactive moieties. To assess cytotoxicity, we incubated nanogels with murine fibroblasts for 24 hours and measured the impact of nanomaterial exposure on the cells membrane integrity and metabolic activity.

Intact nanogels (degradable and nondegradable) exhibited limited toxicity to fibroblasts after 24 hours of incubation, while nanogels degraded by 10 mM glutathione in cell culture medium were nontoxic at concentrations up to 2 mg/ml (fig. S7A). Fibroblast membrane integrity was largely unaffected by 24-hour incubation with nondegradable, degradable, or degraded nanoparticles (fig. S7B), indicating that the reduction in metabolic activity observed in fig. S7A was not due to cell lysis. Modification of nanogels with tyramine or N,N-dimethylethylenediamine did not alter their cytotoxicity (fig. S7C), as measured by cell metabolic activity following 24-hour exposure to a dose of 2 mg/ml. Peptide incorporation (fig. S7D) at approximately 2 weight % (wt %) of the dry nanogel weight did not significantly affect nanogel cytotoxicity (fibroblasts, 24-hour exposure, 2 mg/ml dose), as peptide-modified nanogels did not alter the cells metabolic activity.

We then monitored the extent to which nanogel toxicity differed across different cell types (fibroblast, macrophage, and colon epithelial). These were selected as model cell systems for the different cell types that would experience a nanomaterial insult following injection. We recognized that each cell line would interact with the nanogels differently, altering the extent to which the material impairs the cell viability. No significant differences were observed in the cells viability, as determined by metabolic activity or membrane integrity, for degradable, nondegradable, or degraded nanogels at concentrations up to 2 mg/ml (fig. S8). It is noteworthy that we saw a nonstatistically significant trend in macrophage activity, where metabolic activity increased and membrane integrity decreased at the top concentration (2 mg/ml, 24 hours). This does indicate acute toxicity to macrophages at this dose.

We assessed the impact of the nanogels chemistry, through surface modification, on their uptake by different model cell lines. We selected fibroblasts, macrophages, and epithelial cells because they model components of the connective tissue, immune system, and tissues/organs, respectively. Furthermore, by selecting colon epithelial carcinoma (SW-48) cells as the epithelial model, we simultaneously probed the impact of surface modification on preferential uptake by human tumor cells.

Modified nanogels for uptake studies were prepared in the same manner as in previous modification efficiency, therapeutic efficacy, and cytotoxicity studies, except for the addition of a carboxylic acidreactive fluorophore in the modification solution. To make the nanogels fluorescent, we added 5-(aminoacetamido)fluorescein at 0.8 wt % of the dry polymer (for comparison, the tyramine or N,N-dimethylethylenediamine ligand was added simultaneously at 10 wt %) to the modification solution. This fluorophore was conjugated to all of the nanogel formulations, including the unmodified nanogels.

The fluorophore was successfully conjugated to unmodified, TMOD, and DMOD nanogels, although a decreased fluorophore coupling efficiency was observed for DMOD nanogels. We produced calibration curves for all modified nanogel formulations and normalized our subsequent image analyses to the relative slope for each formulation (correction factors: unmodified, 1.27; TMOD, 1; and DMOD, 5.44). We also validated that fluorophore conjugation did not significantly alter the nanogels cytotoxicity by conducting MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] and LDH (lactate dehydrogenase) assays for nanogel exposure to each of the three cell lines at concentrations up to 2 mg/ml. No significant cytotoxicity was observed by either measure at concentrations up to 1 mg/ml (24-hour exposure, all three cell lines) (fig. S9). Consequently, the maximum nanoparticle dose for all uptake studies was maintained at 1 mg/ml.

Even at low doses (less than 40 g/ml, 24-hour exposure), murine macrophages imbibed substantial quantities of all three nanogel formulations (Fig. 5, A to C). On the other hand, fibroblasts exhibited limited uptake of unmodified and TMOD but took up DMOD nanogels. Human colon epithelial cells took up all three formulations, exhibiting no preference for unmodified or TMOD nanogels, but a 13.4-fold increase in uptake when exposed to DMOD nanogels (relative to unmodified nanogels, 250 g/ml, 24-hour exposure). Representative images, visualizing nanogel uptake by each of the three cell lines, are given in fig. S10.

The relative uptake was computed by normalizing the green fluorescent protein (GFP) (nanoparticle) signal to the slope of the calibration curve and then normalizing that value to the 4,6-diamidino-2-phenylindole (DAPI) (cell nucleus) signal. Note that the y axis quantities differ between plots, as the DMOD nanogels were uptaken in significantly greater quantity than TMOD or unmodified nanogels. (A to C) Relative uptake of unmodified, TMOD, or DMOD fluorescent nanogels by each cell line, as a function of dose (24-hour exposure). (D to F) Kinetic uptake of unmodified, TMOD, and DMOD nanogels (400 g/ml dose). Representative images for each plot are given in fig. S10 (all panels, n = 4, mean SEM).

These dose-response results demonstrated that, while nanogel modification with N,N-dimethylethylenediamine generally increased uptake, the extent to which uptake was enhanced differed between cell lines. Compared with unmodified nanogels, DMOD nanogels exhibited a 4.5-fold increase in uptake by macrophages, 11.6-fold by fibroblasts, and 17.0-fold by colon carcinoma cells (250 g/ml, 24-hour exposure, all differences significant at the P < 0.05 level). This result suggests that the cell-nanomaterial interactions, which promoted uptake and were imparted by the N,N-dimethylethylenediamine moiety, triggered varying degrees of response from different cells. Furthermore, TMOD nanogels were uptaken similarly to unmodified nanogels. Tyramine modification led to a 21% decrease in uptake by macrophages, 31% decrease by fibroblasts, and 3.8% increase by colon carcinoma cells, none of which were statistically significant. This confirmed that a surface modification that imparts environmental responsiveness or alters therapeutic partitioning does not necessarily also enhance cell uptake.

Image analysis revealed that the nanogels interacted with each cell line in a different spatiotemporal manner. Nanogels did not interact substantially with fibroblasts, and when they did, they colocalized primarily with the cell membrane. Macrophages rapidly internalized the nanogels, with images demonstrating cytosolic colocalization in as little as 30 min. In the case of colon epithelial cells, nanogels first associated with the cell membrane, which preceded uptake. DMOD nanogels associated with the colon epithelial cells membranes and were internalized more rapidly than unmodified and TMOD nanogels (fig. S10).

Kinetic analyses of nanogel uptake further clarified the differences in nanogel uptake within cell lines and between formulations (Fig. 5, D to F). For precision medicine applications, we want to ensure that target cells (i.e., colon cancer cells) internalize the nanomaterial prior to complete therapeutic elution or clearance by off-target cells (i.e., fibroblasts or macrophages). It is relevant to recall that, depending on the particular surface modification and pH environment, the majority of the loaded methylene blue was eluted in less than 4 hours. Therefore, a formulation that rapidly associates with, and facilitates uptake by, target cells will enhance cytosolic delivery of the payload.

Murine macrophages took up all three nanogel formulations with near zero-order kinetics for the first 6hours. On the other hand, colon carcinoma cells exhibited a rapid cell-nanoparticle association [i.e., a spike in the green fluorescent protein (GFP)/4,6-diamidino-2-phenylindole (DAPI) signal in the first 15 min to 2hours], followed by a plateau in the signal intensity. Nanogels did not associate with the membrane or cytosol of murine fibroblasts until 24 hours of exposure. We looked specifically at nanogel uptake within the first 2 hours of dosing, as this is when majority of the methylene blue elution occurred in our drug release studies (at pH 7.4, 41% released by unmodified, 64% released by TMOD, and 91% released by DMOD). To compare the nanogel uptake at 2hours across cell lines, we computed the relative uptake as the ratio of the 2- and 24-hour uptake (400 g/ml dose). A two-way analysis of variance (ANOVA) revealed that formulation accounted for only 11.1% of the total variation in relative uptake (not significant), whereas the cell line identity explained 38.6% of the total variation in relative uptake (significant at the P < 0.001 level). Consistent with the dose-response study presented above, DMOD uptake by colon cancer cells was 12.4 times greater than that of unmodified nanogels. Tyramine modification did not significantly affect the extent of nanogel uptake by any cell line.

The rapid association and uptake of DMOD nanogels by colon cancer cells is particularly interesting, as it suggests that this particular surface modification could enhance specific drug delivery to target tumor cells. However, as these experiments were conducted in homogeneous, static cell cultures, we are unable to conclude whether this preference for colon tumor cells would translate to in vitro coculture or in vivo models.

One advantageous therapeutic quality of DMOD nanogels was their ability to act as an intrinsic reducing agent and, subsequently, act as centers for gold nanoparticle precipitation. DMOD nanogels with the three highest degrees of modification (0.78:1 DMOD or greater) were able to reduce gold chloride successfully, forming nanogel-coated gold nanoparticles. Nanogels with lesser quantities of N,N-dimethylethylenediamine did not form gold nanoparticles. Analysis of the composite nanogels absorbance spectrum (Fig. 6A) revealed that the conjugates absorb visible light strongly, with a maximum absorbance wavelength of 536 nm. In the transmission electron microscopy (TEM) images presented here (Fig. 6B), the gold nanoparticles are visible as dark circular regions within the nanogel bulk. Some, but not all, of the nanogels contained gold nanoparticles after the precipitation reaction.

Gold nanoparticles were precipitated in DMOD nanogels. DMOD gels with a 0.39:1 ratio of N,N-dimethylethylenediamine:methacrylic acid or less were unable to facilitate gold nanomaterial formation. (A) Absorbance spectra of composite nanogels containing gold nanoparticles. (B) Transmission electron micrographs of gold nanomaterials within 3.1:1 DMOD nanogels. Arrows point to gold nanoparticles. (C) Proof of concept for the composite nanogels ability to transduce visible light (=532 nm) into heat. DMOD (3.1:1) nanogels with gold nanoparticles effectively and rapidly heated a 1 PBS suspension. (D) Concentration-dependent photothermal activity of 3.1:1 DMODgold nanoparticle composites (n = 4, mean SD).

DMOD nanogelgold nanoparticle composites (3.1:1) were suspended at various concentrations in 1 PBS and were irradiated with a 532-nm laser at 200 mW. Within 30 s, the PBS suspension reached an equilibrium temperature (Fig. 6C), while the heat rapidly dissipated when the laser was turned off. Nanogels alone, in the absence of precipitated gold nanoparticles, did not heat the surrounding medium when irradiated with the same laser, indicating that the gold nanomaterials were acting as a transducing element. The heat generated by laser irradiation increased with nanoparticle concentration, with a maximum heating of 10.30.20C by a nanoparticle-in-nanogel suspension (1 mg/ml) (Fig. 6D).

Next, we demonstrated the feasibility of peptide and protein coupling to the base nanogel platform. Peptides can be used to impart specific biological behaviors, including molecular recognition, cell targeting, cell penetration, and endosomal escape. Bioactive proteins can contribute enzymatic activity to the otherwise inert network or be used as a molecular recognition unit for targeting applications.

Two independent peptide conjugation reactions were explored: one for coupling cysteine-containing peptides via a thiol-maleimide reaction and a second for coupling the N-terminal amine or pendant lysine groups to carboxylic acids in the nanogel network. Five diverse, cysteine-containing peptide sequences were selected to sample a diverse array of peptide properties (two cationic, two anionic, and one electrically neutral at physiological pH, all water soluble). These peptides were previously identified by the authors as candidates for trypsin recognition in physiological fluids. In the present study, they were used as model oligopeptides to optimize a generalized nanogel-peptide conjugation strategy and conclude relationships between a peptides formal charge and its coupling efficiency. In a two-step conjugation schema (Fig. 7A), we first coupled a maleimide-terminated linker molecule to the nanogels via carbodiimide-mediated coupling (pH 4.5). After 2 hours, we adjusted the nanogel suspension pH to 7.0 to favor the thiol-maleimide click reaction with the cysteine-containing peptides, as opposed to any aminecarboxylic acid side reaction (i.e., those between the nanogels carboxylic acid and the peptides N terminus, or peptide dimerization via the C and N termini of multiple peptides).

(A) A thiol-maleimide click reaction effectively conjugated cysteine-containing peptides to the nanogel network. (B) A carboxylic acidamine reaction linked the peptides N terminus with the carboxylic acidcontaining nanogels. (C) Differential incorporation of diverse peptides was explained by their net charge at physiological pH. (D) Nanogel conjugation at 2 wt % did not significantly alter the nanogel diameter or zeta potential. (E) Peptide content in the final conjugate product can be readily tuned by altering the peptide feed concentration via reaction scheme (B). (F) Horseradish peroxidase (HRP) and wheat germ agglutinin (WGA) were incorporated into nanogels with 56.4 and 81% efficiency, respectively. (G) HRP retained 66.533% of its activity upon conjugation to the nanogel platform, as evidenced by the ability of HRP-nanogel conjugates to convert TMB substrate. (H) WGA-NP conjugates retained native WGA activity, as they bound and stained the cell membrane of L929 murine fibroblasts (blue, DAPI stain of nucleus; red, WGA-NP conjugates) (C to F, n = 3, mean SD; G and H, n = 3, representative data).

We were successful in conjugating all five peptides to the network, demonstrating the feasibility of conjugating diverse peptide ligands to the platform. Peptide content, within each nanogel network, was quantified with a Micro BCA colorimetric assay. Cationic (FAHWWC and HAHWEC) and electrically neutral (CDHFAI) peptides were incorporated with nearly complete efficiency (theoretically complete incorporation was 2% of the dry weight). On the other hand, anionic peptides were incorporated with lesser efficiency (43.78.5% and 50.98.6% for CDNWQY and ADCFLQ, respectively) (Fig. 7C). This highlighted the effect of peptide formal charge, which influences its equilibrium partitioning in the nanogel phase during the conjugation reaction, on efficient coupling. The extent of nanogel decoration with anionic peptides was increased by elevating the concentration of the anionic peptide in the coupling reaction, but is still significantly less efficient than the coupling of neutral and cationic peptides. Nanogel decoration with peptide, at 2 wt %, did not significantly alter the nanogels size or zeta potential (Fig. 7D).

In a separate bioconjugation schema, the peptides were linked directly to the nanogel network via a reaction between the peptide N terminus and pendant carboxylic acid groups (Fig. 7B). Again, conjugation of a cationic peptide (HAHWEC) was efficient, as the quantity of the peptide within the network was readily controlled by modulating the peptide concentration in the coupling reaction (Fig. 7E). Our model anionic peptide (CDNWQY) was incorporated into the nanogels, but with a lesser efficiency (62.715.0%, depending on the peptide concentration in the modification solution).

Wheat germ agglutinin (WGA) and horseradish peroxidase (HRP) were conjugated to nanogels via carbodiimide-mediated coupling, with 2 wt % protein in the modification reaction. These model proteins were selected, as they are commonly applied for immunohistochemistry and biosensing applications, respectively. As a result of selecting these two protein targets, we had methods for verifying the retention of protein activity following conjugation to the nanogels. Furthermore, the result is applicable to other proteins that have affinity for extracellular targets (similar to WGA) or catalyze small-molecule conversion (similar to HRP). Each protein was incorporated successfully (Fig. 7F) and retained its bioactivity after conjugation. HRP activity was quantified by the colorimetric determination of enzymatic conversion of 3,3,5,5-tetramethylbenzidine (TMB) substrate. Standard curves were generated for TMB conversion as a function of HRP concentration (free HRP or HRP bound covalently to nanogels). By comparing the conjugated HRP activity to that of free HRP at the same concentration, we determined that 66.533% of the HRP activity was conserved (Fig. 7G). This provided evidence that our nanogels acted as scaffolds for retaining and presenting bioactive HRP to the surrounding environment.

WGA activity was assessed by determining the effectiveness with which WGA-labeled nanogels labeled fibroblast cell membranes. Fibroblasts were selected because unlabeled nanogels neither associate with fibroblast cell membranes nor are uptaken by fibroblasts within 2 hours, as quantified in Fig. 6 and illustrated in fig. S10. Therefore, colocalization of nanogels with the fibroblast membranes, or uptake into the cytosol, is due to the membrane-targeting activity of WGA. As shown in Fig. 7H, the WGA-labeled nanogels (red) colocalize with the cell cytosol, indicating that the conjugated WGA facilitated cell-nanogel interactions and subsequent uptake.

Here, we documented a modular, tunable nanogel platform for therapeutic applications. P(AAm-co-MAA) nanogels were decorated with numerous amine-containing ligands (i.e., small molecules, peptides, and proteins) and retained the ligand bioactivity (i.e., intrinsic reducing ability, pH sensitivity, hydrophobicity, molecular recognition characteristics, and enzymatic activity). We tuned the extent of ligand decoration by modulating the characteristics of the modification reaction and yielded a range of therapeutic capabilities, including cell targeting, enhanced nanomaterial uptake, intelligent drug delivery, and photothermal therapy.

In its present form, unmodified P(AAm-co-MAA) nanogels are suitable for loading high weight fractions of hydrophilic, cationic therapeutics. A suitable initial chemotherapeutic agent will be 5-fluorouracil, which is used to treat a number of cancers including colorectal cancer. These unmodified nanogels exhibited sustained therapeutic delivery for greater than 6 hours. Tyramine-modified nanogels were responsive to the pH environment and, as a result, delivered methylene blue more rapidly in acidic than neutral buffer. N,N-dimethylethylenediaminemodified nanogels were amphoteric in nature, eluted methylene blue the most rapidly of the tested formulations, and increased nanogel uptake by colon cancer cells.

Gold nanoparticle precipiation enabled photothermal therapy. Following DMOD nanogel accumulation in tumor sites, excitation with a focused green laser would heat the tumor tissue. Previous studies using gold nanoparticles for photothermal therapy applications have demonstrated efficacious heating using green light (28, 29). However, our platform combines photothermal therapy and chemotherapeutic delivery in a new, modular manner. In the future, platform functionalization with targeting peptides, monoclonal antibodies, or other targeting molecules could further enhance nanogel targeting and cell uptake.

As presented in Introduction, research on treating cancer with multiple therapeutic modalities is increasing in prevalence (30, 31). This platform technology, with its highly tunable nature, is amenable to delivering multiple chemotherapeutics and facilitating combination therapies, each with precise targeting or environmental responsiveness. Calibrated combinations of modified and unmodified nanogels in a single regimen could produce new physical distributions and release profiles of therapeutic agents in the future.

In addition to demonstrating the use of a new platform technology, we introduced multiple new characterization methods, which will be of utmost use to researchers developing nanoscale devices for precision medicine. In particular, while QCM methods have been used previously to study the deposition of nanomaterials on solid surfaces or the interfacial interactions governing monolayer self-assembly (3234), the use of QCM to study swelling and biodegradation of nanogel materials is novel. Similarly, while nanomaterial internalization by cells has been an active area of research, using methods such as confocal microscopy and flow cytometry (35, 36), we developed a high-throughput microplate assay for nanogel uptake. As a direct result of having this new analysis tool, we were able to screen the dose-dependent and kinetic uptake of our modified nanogels by three relevant cell lines.

In conclusion, we engineered a new nanogel platform, which is modularly tunable for precision medicine applications. We quantified the extent to which nanogel composition altered drug-material interactions for the loading and release of cargo, transduction of external signals, targeting of proteins, and uptake by cells. Our new methods, described herein, will also provide new tools to the drug delivery field to rapidly screen or precisely quantify biological interactions with engineered nanomaterials in the future.

Nanogels were synthesized by inverse emulsion polymerization, as previously optimized by Zhong et al. (16). Acrylamide [75 mole percent (mol %)], methacrylic acid (22.5 mol %), and methylene bisacrylamide (2.5 mol %) were dissolved in water at 42 wt %. This aqueous phase (2.762 ml) and N,N,N,N-tetramethylethylenediamine (50 l; catalyst) were added slowly to a stirring solution of Brij 30 (151.4 mM) and AOT (dioctyl sulfosuccinate sodium salt; 30.3 mM) in hexanes (50 ml) to form a water-in-oil emulsion. This prepolymer emulsion was purged with nitrogen for 20 min to remove dissolved oxygen, and polymerization was initiated by injecting 10 mg of nitrogen-purged ammonium persulfate [100 l of a freshly prepared stock (100 mg/ml) in ultrapure water]. After 2hours, the reaction was stopped by opening the round-bottom flask to air, and the nanogels were purified by precipitation in ethanol (three times) followed by either ionomer collapse or dialysis against a water:ethanol gradient.

In ionomer collapse, the nanogels were suspended in 0.5 N sodium hydroxide and precipitated with the addition of a threefold volume excess of acetone. Precipitated nanogels were collected by centrifugation (3200g for 5 min), and the collapse procedure was repeated an additional four times. In gradient dialysis, nanogels were suspended in a 50:50 water:ethanol mixture and dialyzed against a decreasing water:ethanol gradient for >5 days with twice-daily dialysate change. Nanogels purified by both ionomer collapse and gradient dialysis were then exchanged into ultrapure water by dialysis. All purified nanogels were lyophilized and stored at room temperature.

For studies involving degradable nanogels, synthesis was conducted in the manner described above, with N,N-bis(acryloyl)cystamine substituted for methylene bisacrylamide. N,N-bis(acryloyl)cystamine is a biodegradable cross-linker that is labile via reduction of its disulfide bond. Cross-linker comprised 2.5 mol % of the monomer feed, and the masses of acrylamide and methacrylic acid were adjusted such that total monomer concentration remained 42wt % in water.

Nanogels were suspended at 10 mg/ml in 1 PBS and adjusted to pH 7.4. Then, 0.5 ml of nanogels and 0.5 ml of DTT or glutathione (20 mM in 1 PBS, pH 7.4) were mixed in a polystyrene cuvette, immediately after which light scattering measurements were recorded. Measurements were recorded using a Zetasizer Nano ZS (Malvern) with a manual attenuation (Attn, 7), measurement position (4.65 mm), and measurement time (10s). Measurements were taken repeatedly for 50 min. In each interval, a hydrodynamic diameter and count rate were recorded. Because count rate trends with the number of particles in solution (37), the count rate at a given time, normalized to the initial count rate, provided a measure of the degree of degradation.

QCM studies were conducted using QSense E4 QCM-D (Biolin Scientific). Uncoated gold sensors were washed in a 5:1:1 volume ratio of ultrapure water, ammonia hydroxide (25 volume %), and hydrogen peroxide (30 volume %) at 75C for 5 min. The sensors were then washed with an excess of water and an excess of ethanol and were dried under nitrogen. Immediately prior to experimentation, clean sensors were treated with ultraviolet/ozone for 10 min.

All experiments were conducted in their entirety at 37C and a flow rate of 0.200 ml/min. A stable baseline for the sensors was achieved by flowing 1 PBS for at least 10 min. Then, the sensors were coated with an amine-terminated monolayer through the addition of cysteamine HCl (10 mg/ml in 1 PBS). Nanogels activated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (twofold molar excess relative to MAA) were flowed over the modified sensor at 2 mg/ml. After a stable coating was obtained (as evidenced by no further fluctuation in the frequency or dissipation), 1 PBS was flowed over the sensor to wash away unreacted polymer and remaining catalyst. The change in resonance frequency, as a result of nanogel coupling, was recorded as a measure of the bound nanogel mass.

Nanogels were swelled in a series of buffers differing in ionic strength (PBS buffers at pH 7.4 diluted to 5, 2, 1, 0.5, 0.1, and 0.01 with ultrapure water) as well as 1 PBS buffer adjusted to different pH values (3, 5, 7, 9, and 11). The purpose of these swelling steps was twofold: to quantify the responsiveness of nanogels to environmental conditions and to verify that the nanogels are behaving as expected despite their immobilization. The frequency and dissipation values were monitored to quantify the nanogels water uptake or expulsion in each buffer condition, as well as to determine the viscoelastic properties of the nanogel layer.

Nanogels were degraded by flowing a 10 mM DTT solution over the nanogel-modified sensors. The frequency and dissipation values were monitored to determine the mass loss during degradation as well as probe changes in viscoelastic properties that indicate the degradation mechanism (i.e., bulk degradation or surface erosion). In each case, the mass immobilized or adsorbed was quantified by the Sauerbrey equationm=Cfnwhere m is the mass adsorbed or immobilized, C is a constant that depends on the intrinsic properties of quartz [for a 5-MHz crystal, C = 17.7 ng/(cm2 Hz)], and n is the overtone number (i.e., 3 and 5). The relative mass was computed by normalizing the change in mass, due to swelling or degradation, to the mass of nanoparticles immobilized (32, 38). It is also equal to the ratio of the frequency changesmdegradationmimmobilized=fdegradationfimmobilized

Purified, dried nanogels were suspended in 10 mM MES buffer and adjusted to pH 4.50.05. Tyramine or N,N-dimethylethylenediamine was dissolved in water at 25 mg/ml. EDC hydrochloride was dissolved immediately prior to use in MES buffer at 56 mg/ml. Each reaction was composed of 5 ml of nanogels (50 mg), 1 ml of EDC solution (56 mg of EDC, a 2:1 molar ratio EDC:MAA by original synthesis feed, and 0.8:1 molar ratio EDC:acid subunit when confirmed by titration analysis), and a variable volume of tyramine or N,N-dimethylethylenediamine (1.6 ml for the highest degree of modification and cut by half for each subsequent reaction). The highest modification ratio (by moles) tested was 2:1 tyramine:MAA and a 3.1:1 N,N-dimethylethylenediamine:MAA. In each highest modification case, the ligand concentration was 80% that of the nanogels by mass. As a control, nanogels were subjected to the reaction conditions (MES buffer, pH 4.5, 56 mg of EDC) in the absence of ligand. Modified nanogels were purified by dialysis against ultrapure water (>72 hours, frequent water changes). Reactions were completed in duplicate.

The physicochemical properties of nanogels were quantified by attenuated total reflectanceFTIR spectroscopy (Nicolet iS10 FTIR Spectrometer; Thermo Fisher Scientific), DLS, and zeta potential measurement (Zetasizer Nano ZS; Malvern). Dried nanogels were pressed in contact with a germanium crystal, and the IR absorption spectrum was recorded from 4000 to 675 cm1. All presented spectra are the average of 64 measurements. All spectra were normalized such that the magnitude of their largest peak was 1U, and their baseline was set at zero. All DLS measurements, unless otherwise stated, were obtained at a nanogel concentration of 2 mg/ml in 1 PBS, adjusted to pH 7.4. All zeta potential measurements, unless otherwise stated, were taken at a nanogel concentration of 2 mg/ml in 5 mM sodium phosphate buffer, adjusted to pH 7.4.

The extent of modification with tyramine, N,N-dimethylethylenediamine, or 5-(aminoacetamido)fluorescein was quantified by potentiometric titration. Ten milligrams of modified or unmodified nanogels was suspended in 60 ml of 5 mM potassium chloride buffer. The suspension was adjusted to pH 10 with 1 N sodium hydroxide to completely deprotonate pendant methacrylic acid groups. The solution was titrated through the equivalence point with 0.01 N hydrochloric acid (HCl) using an autotitrator (Hanna HI901C). We titrated the nanogel suspensions from a basic-to-acidic environment to ensure that the nanogels were swollen during the entirety of adjustment to equivalence. The equivalence point for methacrylic acidcontaining nanogels was consistently at pH 4.8.

We assume that at equivalence (pH 4.8), exactly half of the acid moieties are protonated. We also assume that at a pH three points above equivalence (pH 7.8), 0.1% of the acid groups are protonated. The volume of 0.01 N HCl needed to adjust pure 5 mM KCl buffer from pH 7.8 to pH 4.8, as well as the volume needed to adjust each nanogel suspension the same increment, was recorded. Using these measurements and the stated assumptions, we calculated the mass fraction of methacrylic acid groups usingmMAAmnanogels=(10.499)(VsuspensionVbuffer)NtitrantMWMAA1mnanogelswhere mnanogels was 0.010 g, Ntitrant was 0.01 M, the molecular weight of methacrylic acid (MWMAA) is 86.06 g/mol, and both volumes were measured in liters.

The extent of nanogel functionalization with peptides or proteins was quantified using a Micro BCA colorimetric assay (Thermo Fisher Scientific), as described previously (39). Modified or unmodified nanogels, suspended at 2 mg/ml in 1 PBS (pH 7.40.05), were combined at an equal volume ratio with Micro BCA working reagent and mixed for 2 hours at 37C (constant mixing). The absorbance of the reduced supernatant (=562 nm) was used to quantify the suspensions peptide concentration, relative to standard curves generated for each pure peptide or protein. The background absorbance of unmodified nanogels under the same testing conditions was subtracted from each measurement.

L929 murine fibroblasts, RAW 264.7 murine macrophages, and SW-48 human colorectal epithelial carcinoma cells were chosen as model cells to properly assess nanomaterial interactions with model connective, immune, and epithelial tissues that would interact in vivo. All cells were cultured in T-75 tissue culturetreated flasks and were used at passages ranging from 6 to 20.

Cells were incubated in a sterile 37C, 5% CO2 environment. Culture medium for all three cell lines was phenol redcontaining high-glucose Dulbeccos modified Eagles medium (DMEM), supplemented with 10% fetal bovine serum (FBS), 2 mM l-glutamine, and 1% penicillin-streptomycin. Cells were passaged once they reached 80 to 90% confluency. Experiment medium for all three cell lines was phenol redfree, high-glucose DMEM with 2% FBS, 2 mM l-glutamine, and 1% penicillin-streptomycin. Experiments were conducted when cells reached 50 to 70% confluency.

For all cell assays, L929 and RAW 264.7 cells were seeded in tissue culturetreated 96-well plates at a density of 10,000 cells per well. SW-48 cells were seeded in similar plates at 25,000 cells per well. Cells were given a minimum of 24 hours to attach and reach 50 to 70% confluency before cytotoxicity, or nanogel uptake assays were performed.

Methylene blue was selected as a model hydrophilic, cationic therapeutic. Methylene blue is a photosensitizer and was selected because of its similarity in hydrophilicity and ionization to hydrophilic chemotherapeutics (i.e., 5-fluorouracil), as well as its compatibility with our hydrophilic, anionic nanogels. Methylene blue was loaded into modified and/or unmodified nanogels by equilibrium partitioning in ultrapure water. For loading experiments, methylene blue (2 mg/ml) and purified nanogels (2 mg/ml) were mixed for 15 min in distilled water. Drug loading was quantified by removing a sample (500 l) and separating the unbound drug by ultrafiltration (Sartorius Vivaspin 500; 300,000 MWCO). The unbound methylene blue was quantified by absorbance (=590 nm) relative to a standard curve. Loaded or partitioned methylene blue was quantified usingQ=(C0Ce)Vmwhere Q is the mass ratio of loaded methylene blue to nanogels, C0 is the methylene blue concentration in the loading solution (1 mg/ml), Ce is the unbound concentration of methylene blue (that passed through the filter), V is the volume of the loading solution, and m is the mass of nanogels in the loading solution.

Prior to drug release experiments, unloaded drug was removed by dialysis against ultrapure water (24 hours, 12,000 to 14,000 MWCO). Methylene blueloaded nanogels [10 ml, nanogels (1 mg/ml), methylene blue (1 mg/ml) in the loading solution, variable methylene blue loaded], still within dialysis tubing, were transferred to 1 PBS solution (400 ml) at pH 4.5 or 7.4 under constant stirring at T=37C. At regular time intervals (15 min, 30 min, 1 hour, 1.5 hours, 2 hours, 4 hours, 8 hours, 24 hours, and 28 hours), samples were taken both from within and outside the dialysis tubing. For samples drawn from within the dialysis tubing, loaded and released methylene blue were separated by ultrafiltration. The dialysate was exchanged for fresh buffer at the 2-hour time point and each time point thereafter to simulate drug metabolism. This dialysate exchange ensured that a concentration gradient (between the nanogel and solution phases) was maintained to facilitate complete methylene blue elution. The total released drug was quantified for the first time point asmreleased=Vwithin tubingCMB,within tubing+VdialysateCMB,dialysatewhere the volume parameters describe the total volume within and outside of the dialysis tubing, respectively, and the concentration parameters capture the released methylene blue present in each solution location.

Note that the volume within the dialysis tubing changes with each time point (as sample is depleted) and that released methylene blue within the dialysis tubing eventually dissipates into the dialysate. It is important to correct for these mathematically. For example, at the 30-min time point, the methylene blue released in the 15-min increment from 15 to 30 min was calculated usingmreleased=(Vwithin tubingCMB,within tubing)t=30(Vwithin tubingCMB,within tubing)t=15+(VdialysateCMB,dialysate)t=30(VdialysateCMB,dialysate)t=15

Please note that this equation holds for all future time points as well, changing the respective time indicators, with the one exception that the final term is omitted if the dialysate buffer was exchanged prior to the interval.

Culture medium was removed by plate inversion and replaced with experiment medium containing hydrogel microparticles (0.0005 to 2 mg/ml). In the case of degraded nanogels, the culture medium containing degradable nanogels was spiked with 10 mM glutathione and incubated at 37C for 24 hours prior to the experiment.

Plate layouts were pseudorandomized. To pseudorandomize, we distributed the samples and controls throughout each microplate to ensure that each sample/control was positioned equitably along the plate exterior or within the interior. This controlled for variation in cell proliferation explained by well location within the plate. Following 24-hour incubation, cytotoxicity was quantified via metabolic activity (MTS) and cell membrane integrity (LDH assay).

For MTS assays, the nanogel-containing experiment medium was removed by plate inversion, and cells were washed with 37C Dulbeccos PBS (DPBS) twice to remove adsorbed nanogels and cell debris. Then, 100 l of MTS assay buffer (MTS diluted 1:6 in experiment medium) was added to each well, and relative metabolic activity of each cell sample was quantified by measuring the MTS metabolism (90 min at 37C) within each well, relative to control, as specified by the manufacturer (Promega).

For LDH assays, LDH assay buffer (100 l) was added directly to the cell media containing nanogels and any cell debris. The relative membrane integrity was calculated by measuring the LDH activity (excitation, 560 nm; emission, 590 nm) according to the following relationRelative membrane integrity=100100sampleblankmax releaseblankwhere the sample measurement is the fluorescence of the treatment media with LDH assay buffer, the blank is the cell culture medium and assay buffer without cells, and the max release is the LDH buffer and treatment media after a 20 min incubation of cells with media and 2 l of lysis buffer (Promega).

A high-throughput fluorescence imaging assay was developed for rapidly screening cells uptake of modified and unmodified nanogels. Fluorescently tagged nanogels, with the addition or lack of tyramine or N,N-dimethylethylenediamine, were suspended in phenol redfree DMEM at concentrations ranging from 1000 to 6.25 g/ml. Cells were dosed with either a gradient of nanogel concentration (24-hour incubation) or a set concentration for a range of time (concentration of 400 g/ml).

For concentration-dependent nanogel uptake assays, culture medium was removed from each well by plate inversion and replaced by treatment medium containing suspended nanogels. Cells were allowed to incubate for 24 hours in the presence of nanogels (100 l per well). Following incubation, the nanogels were removed by aspiration, followed by three washes with cold DPBS. Cells were fixed with cold paraformaldehyde solution for 10 min (50 l per well).

For time-dependent nanogel uptake assays, culture medium was removed from each well and replaced with treatment media containing nanogels (400 g/ml) by aspiration in an inverse time manner (i.e., 24-hour time point first, 15-min time point last) (100 l per well). This was scheduled in such a way that all wells reached their end point simultaneously. Nanogel suspensions were removed from the cells by aspiration, and the cells were washed three times with cold DPBS (100 l per well). These cells were also fixed with cold paraformaldehyde solution (50 l per well).

Cells were stained directly in the microplates for fluorescence imaging. Following fixation, each well was washed three times with cold Hanks balanced salt solution (HBSS) (100 l per well). Then, the cell membranes were stained with a WGA Alexa Fluor 594 conjugate solution (3 g/ml) in cold HBSS (15 min) (50 l per well). After three more washes with cold HBSS (100 l per well), the cells were stained with a DAPI solution (1 g/ml) in cold HBSS for 10 min (50 l per well). Each well was washed three times with cold HBSS prior to imaging (100 l per well). Imaging was conducted with 100 l of fresh HBSS in each well.

Cell imaging was conducted at high throughput using a Cytation 3 plate reader (BioTek) with Gen5 software (version 3.04) equipped with DAPI, GFP, and Texas Red filters (DAPI: excitation, 377 nm; emission, 447 nm; Texas Red: excitation, 586 nm; emission, 647 nm; and GFP: excitation, 469 nm; emission, 525 nm) and an Olympus 20 objective. Imaging parameters were optimized to the most fluorescent samples to prevent saturation and were held constant to enable both qualitative and quantitative image analyses between cell lines and treatments [DAPI: light-emitting diode (LED) intensity, 5; integration time, 63; and gain, 0; Texas Red: LED intensity, 10; integration time, 100; and gain, 13.8; and GFP: LED intensity, 10; integration time, 158; and gain, 15]. Four images were taken for each well, and images were preprocessed with a background subtraction step prior to qualitative analysis.

For quantitative analysis, the fluorescence of the whole well was taken for each relevant channel (DAPI, Texas Red, and GFP with gain values of 60, 100, and 120, respectively). To normalize the nanoparticle signal intensity to the cell count, we normalized the fluorescence intensity of GFP to the DAPI channel. The relative nanogel uptake for each cell linecondition pair is given as this ratio.

To precipitate gold nanoparticles within the hydrogel nanogels, chloroauric acid (0.05 wt %) and nanogels (1 mg/ml) were suspended in ultrapure water and mixed (Eppendorf ThermoMixer) at 1000 rpm and 60C for 1 hour. Nanogels with precipitated nanoparticles were used in further experiments without purification. These composite nanogels were characterized by their visible absorption spectra (300 to 1000 nm in 1-nm intervals using a Cytation 3 microplate reader) as well as by TEM (FEI Tecnai Transmission Electron Microscope, operating at 80 kV, cast on carbon-coated grid, and stained with uranyl acetate).

Photothermal experiments were conducted as previously described (40, 41). For photothermal therapy experiments, a 532-nm laser diode (PN156-10.07-0447) was used. This laser wavelength was within the maximum absorbance peak of the gold nanoparticlecontaining nanogels. The nanogels were suspended in ultrapure water at 1 mg/ml, and 1 ml of each nanogel suspension was added to a 24-well microplate. The laser, operated at a power of 200 mW, was focused on a circular area with a 6-mm diameter using a convex lens (Thorlabs Inc.), which was mounted at a 30 angle. Dynamic fluctuation in temperature, within the circular area, was measured using an indium antimonide IR camera (FLIR Systems Inc.).

For peptide modification through a thiol-maleimide click reaction, the nanogels were first modified with N-(2-aminoethyl)maleimide. Purified, dried nanogels were suspended in 10 mM MES at 10 mg/ml and adjusted to pH 4.5. Carboxylic acids were first activated by the addition of a twofold molar excess EDC (relative to MAA content), after which the N-(2-aminoethyl)maleimide trifluoroacetate salt was added. The amount of this linker molecule added was calculated such that if 100% peptide coupling were achieved, then the final peptide concentration would be 2 wt % of the dry nanogel. During this modification reaction, the pH was carefully maintained at 4.5. After 30 min, the pH of the solution was raised to 7.0 with 1 N sodium hydroxide (to terminate the carboxylic acidamine reaction), and the thiol-containing hexamer peptides (FAHWWC, HAHWEC, CDNWQY, ADCFLQ, and CDHFAI) were dissolved in 0.1 PBS at 10 mg/ml, adjusted to pH 7, and added (final peptide concentration of 2 wt % relative to the nanogels). This thiol-maleimide reaction was allowed to proceed overnight at room temperature under constant mixing. The nanogels were purified by dialysis against ultrapure water (12,000 to 14,000 MWCO, >72 hours, frequent water changes).

For peptide modification through a carboxylic acidamine reaction, the nanogels were suspended in 10 mM MES, and pH was adjusted to 5.5. Carboxylic acids were activated with a twofold molar excess (relative to MAA) of EDC. Peptides were dissolved in 10 mM MES at 10 mg/ml and adjusted to pH 5.5. The proper volume of the peptide solution was added to each modification reaction to achieve the desired extent of peptide decoration (i.e., 0 to 10 wt %, relative to the dried nanogels). Nanogels were purified by dialysis against ultrapure water.

Nanogel modification with bioactive proteins was conducted in the same manner as the peptide carboxylic acidamine coupling, except for that WGA Alexa Fluor 594 (Thermo Fisher Scientific) or HRP (Worthington) was dissolved at 1 mg/ml in 10 mM MES and added to the modification reaction at a final protein concentration of 2 wt % (relative to the dried nanogels).

HRP bioactivity within modified nanogels was quantified by its ability to convert TMB substrate, relative to free HRP. Nanogels were dissolved at 2 mg/ml in 1 PBS (pH 7.4 0.05) and diluted 1:8000 for a final concentration of 0.25 g/ml. Lyophilized HRP (Worthington) was also dissolved at 2 mg/ml in 1 PBS and diluted 1:1,000,000 for a final concentration of 0.002 g/ml. A calibration curve for HRP activity was generated via serial dilution with a maximum concentration of 0.002 g/ml. In a 96-well microplate, 100 l of HRP solution or nanogel suspension was mixed with 100 l of TMB substrate solution (Pierce). After 10 min of incubation at ambient conditions, the reaction was stopped by adding 50 l of 1 N sulfuric acid. The reaction product was quantified by visible absorbance at =450 nm.

WGA bioactivity was quantified by its ability to recognize fibroblast cell membrane (via interaction with sialic acid and N-acetylglucosaminyl residues in the membrane). Fibroblasts were seeded in 96-well microplates at 10,000 cells per well and allowed to attach overnight. Cells were incubated in phenol red-free DMEM, supplemented with 2% FBS, containing WGA-conjugated nanogels at 1 mg/ml for 2 hours. As control samples, separate wells were incubated in media alone or media with unmodified nanogels (1 mg/ml) (2 hours). All wells were washed three times with cold DPBS and fixed with cold paraformaldehyde (IC Fixation Buffer; Invitrogen) for 10 min (50 l per well).

After fixation, the nuclei of all cells were stained with DAPI (1 g/ml in cold HBSS, 10 min). The plasma membranes of positive control cells were stained with WGAAlexa Fluor 594 (3 g/ml in cold HBSS, 15 min). After each staining step, all wells were washed three times with cold HBSS (100 l per well).

Fibroblasts were imaged using the fluorescence imaging capabilities of the Cytation 3 microplate reader, equipped with a 20 Olympus objective. So that images could be compared qualitatively, common imaging parameters were used for all images [DAPI (nucleus): LED intensity, 5; integration time, 50; and gain, 0; Texas Red (WGA-nanogels and membrane stain): LED intensity, 10; integration time, 130; gain, 13.6]. Images were processed using Gen5 software (version 3.04), where the background fluorescence was subtracted from each image.

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Synthetic networks with tunable responsiveness, biodegradation, and molecular recognition for precision medicine applications - Science Advances

Self-Assembly Of Retinoid Nanoparticles For Melanoma Therapy | IJN – Dove Medical Press

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Han Liao,1,2 Shan Zhao,1,2 Huihui Wang,1,2 Yang Liu,1 Ying Zhang,1 Guangwei Sun1

1Scientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, Peoples Republic of China; 2University of Chinese Academy of Sciences, Beijing 100049, Peoples Republic of China

Correspondence: Guangwei SunScientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023, Peoples Republic of ChinaTel/Fax +86-411-82463027Email sungw@dicp.ac.cn

Background: Amphiphilic fusion drugs are covalent conjugates of a lipophilic drug and a hydrophilic drug or their active fragments. These carrier-free self-assembly nanomaterials are helpful to co-deliver two synergic drugs to the same site regardless of pharmacokinetic properties of individual drugs. Retinoic hydroxamic acid (RHA) is a fusion drug of all-trans retinoic acid (ATRA) and vorinostat, a histone deacetylase (HDAC) inhibitor showing synergic effect with ATRA on cancer therapy. Although RHA was synthesized in 2005, its nanoscale self-assembly property, anticancer activity, and possible related mechanism are still unclear.Methods: RHA nanoparticles were observed under transmission electron microscope (TEM). Both in vitro cell viability, colony formation assay, and in vivo xenograft mouse tumor model were employed here to study anticancer activity of RHA nanoparticles. The putative synergic anticancer mechanism of activating retinoic acid receptor (RAR) and inhibiting HDAC were investigated via receptor inhibitor rescue assay and in vitro enzyme activity assay, respectively.Results: RHA could form nanoparticle formation by self-assembly and abrogates growth of several solid tumor cell lines even after RHA nanoparticles washout. However, opposite to our initial hypothesis, pre-treating the melanoma cells with RAR antagonists showed no impact on inhibitory effect of RHA nanoparticles, which suggested that the target of the molecule on melanoma cells is not RAR and retinoid X receptor (RXR). Importantly, RHA nanoparticles inhibited the growth of xenograft tumors without obvious impact on haematological indexes and hepatorenal function of these tumor-bearing mice.Conclusion: Our findings demonstrate the promise of RHA nanoparticles in treating malignant melanoma tumors with high efficacy and low toxicity.

Keywords: nano-drugs, self-assembly, retinoid, cancer therapy, melanoma

This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License.By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms.

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Self-Assembly Of Retinoid Nanoparticles For Melanoma Therapy | IJN - Dove Medical Press

Nanomanipulator Market Upcoming Trends, Revenue, Key Manufactures and Competitive Analysis till 2025 – One Digi Click

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Los Angeles, United State, Oct 01, 2019 QY Research provides an authentic report about the global Nanomanipulator Market. It includes market figures, both historical as well as estimates. The research report details the milestones that the global market for Nanomanipulator has achieved and discusses the potential opportunities for the players operating in the market. The publication titled Nanomanipulator includes Porters five forces analysis and SWOT analysis to give its readers a holistic outlook.

The report further explains the nature of competition and its impact on the suppliers and buyers; while the latter explains their strengths, weaknesses, opportunities, and threats. The research report has been compiled using primary and secondary research methodologies to give the readers an unbiased view of the global Nanomanipulator market.

The research report also includes comments and opinions by key market experts to affirm the potential investment opportunities. Analysts have also evaluated the research and development status of these top players and their expansion plans during the forecast period. The section of company profiles includes in-depth scrutiny of the companies and their products, including the ones in the pipeline. The section also discusses the impact of their recent mergers and acquisitions, achievements of the companies so far, and their investments towards product launches.

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Global Nanomanipulator Market: Forecast by Segments

For a better understanding of the market, analysts have segmented the global Nanomanipulator market based on application, type, and regions. Each segment provides a clear picture of the aspects that are likely to drive it and the ones expected to restrain it. The segment-wise explanation allows the reader to get access to particular updates about the global Nanomanipulator market. Evolving environmental concerns, changing political scenarios, and differing approaches by the government towards regulatory reforms have also been mentioned in the research report.

Nanomanipulator Segmentation by Product

Electron Microscope, Scanning Probe Microscope

Nanomanipulator Segmentation by Application

Nanometer Medicine, Biomedical, Machine, Other

Global Nanomanipulator Market: Competitive Landscape

Top Players operating in the global Nanomanipulator market are:

In this section of the report, the global Nanomanipulator market focuses on the major players that are operating in the market and their competitive landscape present in the market. The report includes a list of initiatives taken by the companies in the past years along with the ones, which are likely to happen in the coming years. Analysts have also made a note of their expansion plans for the near future, financial analysis of these companies, and their research and development activities. This research report includes a complete dashboard view of the global Nanomanipulator market, which helps the readers to view an in-depth knowledge about the report.

Nanomanipulator Market Leading Players

BRUKER, JEOL, THERMO FISHER SCIENTIFIC, GINKGO BIOWORKS, OXFORD INSTRUMENTS, EV GROUP, IMINA TECHNOLOGIES, TORONTO NANO INSTRUMENTATION, KLOCKE NANOTECHNIK, KLEINDIEK NANOTECHNIK

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Global Nanomanipulator Market: Research Methodology

QY Research has gathered the data from various sources such as secondary and primary research to provide an authenticate result of the global Nanomanipulator market. To validate the data, the team of analysts has gone through the discussion with the panel members by taking their interviews and conducting the market size to analyze the global structure of Nanomanipulator market and getting the feedback from the product manufacturers.

The report also considers the various factors based on secondary sources, market size, key data parameters such as the market positioning of key players in terms of the regional revenue, segmental revenue. Geographic penetration also shows the market potential, market risk, industry trends, and opportunities. Secondary sources mainly include journals, companies annual reports, website, public and paid databases, and press releases.

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QYResearch always pursuits high product quality with the belief that quality is the soul of business. Through years of effort and supports from the huge number of customer supports, QYResearch consulting group has accumulated creative design methods on many high-quality markets investigation and research team with rich experience. Today, QYResearch has become a brand of quality assurance in the consulting industry.

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Nanomanipulator Market Upcoming Trends, Revenue, Key Manufactures and Competitive Analysis till 2025 - One Digi Click

First-of-its-kind trial in ALS spurs hope for brutal disease – BioPharma Dive

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Until the day she dies, Sandy Morris is hunting for a comma and a clause.

In May 2018, the 53-year-old mother of three learned she has ALS, a diagnosis made even more devastating by a somber directive to get her affairs in order period. The finality speaks to the absence of an effective treatment, yet alone a cure, for the neurodegenerative disease also known as Lou Gehrig's disease.

Since the diagnosis, Morris has sought to give future ALS patients some measure of optimism, amending the 'get your affairs in order' instruction to add 'but, there's something worth looking into.' While medical advances in areas like gene therapy show potential to change the course of other potentially fatal diseases, a disease-modifying treatment in ALS has proved elusive.

"In cancer, you have a shot," Morris said in an interview. "Not always a great shot, but you have one. In this land, everybody is just like, 'We don't have anything.'"

But now, Morris said she has reason for hope, stemming from a first-of-its-kind study in ALS that will test five experimental drugs simultaneously in a platform trial. Five drugmakers selected for the trial were announced last week.

After years of work, the broader promise of these adaptive trials, which allow more flexibility to make changes throughout, is beginning to be put into action. This trial aims to accelerate the pace of ALS research and may prove to be a template for other rare diseases as well, experts said.

The design offers advantages for each key stakeholder in drug development: patients have a lower probability of receiving placebo due to the study's shared control arm; companies get quicker topline results at lower cost; and researchers talk of the study as an "endpoint engine" that could advance their scientific understanding of the disease.

Formidable challenges remain in the way of broader adoption, particularly among leaders of the drug industry. The ALS study uses experimental treatments from small biotechs instead of industry giants, which can afford to prioritize control and pay for their own studies instead.

Even so, it's an exciting advancement in ALS research. Morris, who is also a leader of the patient group I Am ALS, said the trial can bring hope to future patients, even if it comes too late to help her.

"It's not going to save me. I'm not in time for any of this," Morris said. "But, dammit, we have to make it better for the rest."

Merit Cudkowicz understands speed. After 25 years specializing in ALS research, Cudkowicz is fully aware of how ALS changes a patient's life, shortening their life expectancy from decades to just years. The disease has a roughly three-year median survival.

Merit Cudkowicz

Massachusetts General Hospital

That life-changing diagnosis came in May 2018 for Sean Healey, then the CEO of an asset management firm worth more than $8 billion. After resigning from that post, Healey connected with Cudkowicz as he searched for treatment options and realized there was an opportunity to accelerate the pace of research.

Just six months from his diagnosis, Healey raised $40 million to launch a research center at Massachusetts General Hospital. Now, slightly less than a year after being established, the Sean M. Healey & AMG Center for ALS Research is aiming to start the platform trial in the first few months of 2020.

And earlier this month, the Healey Center announced the first five therapies to be tested in the study, including drugs from Biohaven Pharmaceutical and Ra Pharmaceuticals.

Cudkowicz, who leads the study, as well as the Healey Center and Mass General's neurology department, said in an interview the trial aims to have 160 patients for each of five treatments. The primary outcome will be whether or not the drug boosted a functional rating score for ALS after six months. One placebo group will be shared for all treatment arms, and more drugs can be added as the study continues. (Cudkowicz cautioned the plan is nearing finalization with the Food and Drug Administration, so specifics could still change.)

Moving quickly is the trial's aim. Cudkowicz called the study "an endpoint engine" that can help create better outcome measurements, including future surrogate markets, pushing ALS research forward. She estimated topline results to come 12 to 18 months after the study begins, depending on enrollment speed.

But those goals are steps toward the actual mission: developing a cure, Healey wrote in an email to BioPharma Dive.

"Of course, we all understand that the most meaningful measure of success will be the development of effective treatments and ultimately a cure," Healey wrote. "I am convinced that the Platform Trial, along with other initiatives we are supporting, will substantially accelerate the achievement of this ultimate goal."

For the first five biotechs, the decision to participate was a no-brainer. The center and other groups are footing the majority of the cost, leaving the companies' primary expense to simply provide their drug.

Out of about 30 applicants, the Healey Center selected five drugs for the study, most notably Biohaven's verdiperstat and Ra Pharma's zilucoplan. The trial also will test therapies from Implicit Bioscience, Prilenia Therapeutics and Clene Nanomedicine.

Cudkowicz said the typical applicants were small biotechs that "have great ideas but not deep pockets."

The expense of clinical trials limits the ability for biotechs to run multiple studies simultaneously across a range of indications. Instead, companies typically focus on a lead indication, with others following in succession.

"As a small company, we live and die by being able to run efficiently and test our hypotheses," said Irfan Qureshi, Biohaven's vice president of neurology.

For instance, Biohaven's verdiperstat is in Phase 3 testing for multiple system atrophy, and Ra Pharma's zilucoplan is focused on a different neuromuscular disorder called generalized myasthenia gravis.

"We wanted to do the study anyway, but to be honest, we probably wouldn't have gotten to it for years if this had not come along," Ra Pharma CEO Doug Treco said in an interview.

While smaller biotechs have bought into the platform trial and its efficiencies, missing from the list of initial participants are industry leaders. With a market value of about $2 billion, Biohaven is the largest company involved.

Cudkowicz noted larger drugmakers have shown interest in roundtable discussions that shaped the trial's design, and there's always potential to add additional therapies after the trial starts.

"We're talking to the other ones like Biogen and Sanofi," she said. "They are interested and came to these meetings, but they have the money to do it on their own."

But the main hang-up for these companies is ceding control, said Scott Berry, a senior statistical scientist and co-founder of Berry Consultants, which worked on the ALS platform trial.

"All these companies have people that this is what they do for a living they run trials, and they know how to do it," Berry said. "To hand your drug to somebody else and you don't have control over making sure that happens is uncomfortable for companies, and it's different than what they usually do."

Additionally, with lengthy protocols and many moving parts, the trials are complex and typically require extensive consulting to get off the ground. Running a multi-arm study also brings statistical pitfalls that can make it harder to interpret results.

Beyond ALS, adaptive trials have been started in breast cancer, Alzheimer's disease and glioblastoma. Experts say they anticipate other rare diseases as logical future targets for these studies.

"As these start to get developed and people see them, in most rare diseases there will be people jumping on board," predicted Berry.

Biohaven's Qureshi added that once companies are willing to experiment beyond typical drug development, these trials could be particularly attractive for rare diseases by easing enrollment concerns where there are "not patients growing on trees," he said.

But the ultimate test to get the industry's full attention, Qureshi said, would be such a study yielding an approved drug.

In the meantime, platform trials appear here to stay. Janet Woodcock, the long-time leader of the FDA's drug review center, has been an influential supporter for these study designs. Just this month, the agency finalized guidance on ALS research that advised companies to consider adaptive trial designs.

Earlier this year, Woodcock told BioPharma Dive she believes these types of studies will gain ground as patients become more vocal about how trials are conducted.

Morris said the platform trial has struck her with its compassion. While no patient wants to be put on placebo, it's of particular importance in a disease that progresses as rapidly and severely as ALS, leaving most patients the time to try one study in their life.

She already took her shot, enrolling in a clinical trial that required a three-month observation period before receiving treatment, which then carried a 50% chance of being placebo. "We are humans," she said. "We aren't zebrafish."

Now, she wants to take the baton from "the voices in the graveyard" and pass it onto the next generation. Platform trials may allow her to cover a bit more ground before that hand-off, eventually reaching a day when people might be able to live with ALS, like HIV.

"ALS's day is coming," Morris said.

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First-of-its-kind trial in ALS spurs hope for brutal disease - BioPharma Dive

IN PICTURES: Terry is first person to complete nine-mile swim from Sealand to Dovercourt – Harwich and Manningtree Standard

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IT was third time lucky for a determined fundraiser who has become the first person to complete the nine-mile swim in the North Sea from Sealand to Dovercourt.

Terry Fay, 47, initially set out to complete the challenge of swimming from the offshore platform-turned-principality to Dovercourt Beach on August 18.

Unfortunately, due to some engine issues with the safety boat, the swim did not go ahead as planned on that date.

Terrys second attempt at the ambitious challenge was on September 1, but disaster struck five miles into the swim when the engine overheated.

However, his third attempt, last Sunday, was luckily a success.

Terry, who works for AGI Global Logistics and lives in Dovercourt, started the swim from Sealand at about 10.30am, having planned to come in with the tide to ensure there was enough water over Cork Sands for the safety boat to clear it.

He said: After about two-and-a-half hours of battling the high waves whipped up by the westerly wind, and tide pushing us towards Walton, the tide changed and we then were being pushed towards the shipping lane and Felixstowe.

We stopped a couple of times for me to take energy drinks and have a couple of bananas, and also to warm up a bit my triathlon wetsuit was clearly inappropriate for that amount of time in the water.

We finally arrived into Dovercourt West Beach at about 3.30pm, having swum the nine-mile journey in four hours 38 minutes, to a cheering crowd of about 100 people along the seafront also six kayakers, and a couple of swimmers came to join us for the last few hundred metres.

It was an emotional end to a very long campaign.

In all, Ive swum 55 open water sea miles these past three months, in readiness for - and including - the swim itself.

Terrys swim has raised more than 2,000 for the Royal Marsden Hospital, which helped a close family member of his earlier this year.

Money raised by Terry will go towards the hospitals world-leading cancer centre.

To donate, visit justgiving.com/fundraising/terry-fay6.

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IN PICTURES: Terry is first person to complete nine-mile swim from Sealand to Dovercourt - Harwich and Manningtree Standard

Great Escapes: The Pristine Shores and Year-Round Charm of Denmarks North Sealand – Barron’s

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North Sealand is also known as Nordsjaelland, the King's Coast, and the Danish Riviera. Ian Centrone

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With its iconic architecture, world-renowned culinary scene, and countless cultural lures, its easy to get swept away by Copenhagens storybook charm. But while legions of eager sightseers and well-heeled globetrotters descend upon Denmarks colorful capital city every year, few manage to make it beyond Copenhagens border. Despite the fact that its a relatively small country (especially when compared to its Nordic neighbors like Sweden, Finland, and Norway), there are still plenty of regions worth exploring just outside the city limits.

North Sealand is a prime example, encompassing the northernmost portion of the Danish island of Zealand (which is also home to Copenhagen). Boasting a slew of nicknames, the region is also known as Nordsjlland, the Kings Coast, and the Danish Riviera. But with its quaint villages, coveted beaches, and coastal vibe, this laid-back-but-luxe retreat feels more like Denmarks very own version of Cape Cod.

For generations, in-the-know Danes have flocked to North Sealands pristine shores for a quintessential summer escape (including a long list of royals and other members of Denmarks upper echelon). These days, the dreamy destination is expanding its appeal and attracting visitors all year long, not just during the sweltering high season. Considering its unique combination of historic castles, stunning properties, and stellar gastronomic gems, it isnt hard to understand why. So the next time you find yourself exploring ever-enchanting Copenhagen, be sure to extend your visit with a jaunt up north to experience this idyllic slice of Denmark.

STAY

Anyone who has ever ventured to Copenhagen may already be familiar with the award-winning Hotel Sanders, a five-star boutique property owned by renowned Danish ballet dancer Alexander Klpin. Its sister property, Helenekilde Badehotel, is the star of North Sealand, located in charming seaside town of Tisvildeleje. Built in 1896 as a private summer home by a construction magnate, it was acquired by Klpin in 2001 and underwent a complete renovation in 2008 to update the landmark property while preserving its familial spirit. Today, the winsome retreat is known for its sun-drenched guestrooms, sweeping overlooks of the Kattegat, and dynamic restaurant serving modern Nordic cuisine.

Hotel Hornbkhus is another beloved seaside sanctuary just minutes away from the windswept shores of Hornbk Beach. Originally constructed in 1904, the recently renovated property boasts 36 whimsical rooms that perfectly marry hygge ideals with retro design details. For direct beach access, book a stay at the hotels sister property, Villa Strand, which offers 15 rooms and debuted earlier this year.

DINE

Restaurant Sletten has developed a sparkling reputation that continues to seduce both tourists and locals alike, thanks to its Michelin-star prestige and privileged waterfront view. Located in the idyllic town of Sletten, the restaurant is nestled among former fishermens cottages and serves an inventive a la carte menu of small plates that showcase seasonal local ingredients.

For farm-fresh cuisine, reserve a table at Rabarbergaarden. The limited menu constantly changes based on whatever produce is being harvested from the restaurants organic farm at any given time. Meat and fish dishes are also available throughout the seasons, with all ingredients being sourced from the region. Be sure to explore their extensive wine list, or try a cocktail made with buzzworthy Botanisk Gin (made using a selection of Danish berries and botanicals).

From its bespoke design details to its impeccable menu, Tisvilde Kro is another North Sealand gem that is not to be missed. Danish entrepreneur and multimillionaire Thorvald Stigsen (known for founding travel brands Skygate and Momondo) had the idea to buy a dilapidated building in Tisvilde and save it from demolition by transforming it into a swanky restaurant. After two years of painstaking renovations, the final result proves it was well worth the wait. Just look for the pink building standing on top of the hill.

EXPERIENCE

Art aficionados wont want to miss the Louisiana Museum of Modern Art , located in Humlebaek, just 25 miles north of Copenhagen. As Denmarks most visited museum, the attraction boasts one of the worlds most impressive collections of modern art. Past and present exhibitions include works by visionaries such as Jackson Pollock, Arne Jacobsen, Henri Matisse, Pipilotti Rist, and more. Known for its hot and cold approach (mixing accomplished artists with up-and-coming talents), the museum has been mesmerizing guests since it first opened its doors back in 1958.

For literature-lovers and history buffs alike, be sure to spend some time exploring the port city of Elsinore (also known as Helsingr). Here, visitors can tour the 15th-century Kronborg Castle, which served as inspiration for the setting of William Shakespeare s Hamlet. A designated UNESCO World Heritage Site, the spectacular palace overlooks the resund strait with majesty, marked by its ornate columns, towers, and spires. To see more of North Sealands impressive collection of castles (hence its Kings Coast moniker), pay a visit to Fredriksborg Castle in Hillerd or the Baroque-style Fredensborg Palace and its regal gardens.

Elsinore also houses the M/S Maritime Museum (adjacent to the castle). The collection was established in 1915, but its current subterranean location was conceived in 2013 by the renowned architects of the Bjarke Ingels Group. Built around a dry dock and designed to resemble a ship, the 82,000-square-foot museum contains thousands of artistic works and artifacts related to Denmarks seafaring past.

This writer was a guest of Visit Denmark .

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Great Escapes: The Pristine Shores and Year-Round Charm of Denmarks North Sealand - Barron's

Hefty bill for Sealand farmer who ignored warnings to clear waste off his land – LeaderLive

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A DEESIDE farmer has been landed with a court bill of 3,230 after he ignored warnings to clear waste off his land.

David McCarrick claimed he didnt have time to shift the 450 tonnes of green vegetation and treated wood which had accumulated at Morriston Farm in Sealand. He said he was cash-strapped as he failed to qualify for EU support, so he was prioritising finding a new home for his cattle herd when he quits the tenanted land.

But Deputy District Judge Martin Jackson fined McCarrick 500 with costs of 2,680 and told him: You had notices, but you chose to ignore them.

Prosecuting for Natural Resources Wales, Dafydd Roberts, said McCarrick pleaded guilty three years ago to operating a waste facility without a licence. He received a notice in December 2016 in respect of the waste and received a further notice on May 20 this year asking him to remove it from the land.

When NRW officers visited the Green Lane farm in May last year he told them he was struggling financially and his priority was feeding his herd.

When the officers returned on June 25 this year it was clear the waste had not been removed and it appeared new waste had been piled near to a gate.

There was waste under some power cables which was a major concern in case it caught fire, said the prosecutor.

North East Wales Magistrates Court was told the cost of removing the waste to landfill would be 14,107.

But the court heard McCarrick was due to lose the farm after Flintshire County Council was granted a possession order.

He said he wanted to keep the 100-strong herd so his son could carry on farming and that had been his focus.

Victoria Hanley, defending, said McCarrick accepted the waste had not been removed.

He needs to make arrangements to remove it, but he is in dire financial straits, said the solicitor.

McCarrick, she said, was working on other farms to support his meagre income which had been slashed because of low milk prices and because he was not entitled to the single farm payment.

He has been relying on a charity which has given him pay and corn. But the welfare of his animals has been the priority, said Ms Hanley.

She said the defendant didnt accept hed added to the waste and had started to chip some of it.

McCarrick, 54, admitted failing to remove waste in contravention of a notice served on May 20 this year.

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Hefty bill for Sealand farmer who ignored warnings to clear waste off his land - LeaderLive

Surfer cleans his playground by turning pollution into functional fashion – East Coast Radio

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Mike Schlebach surfs in trash. When he takes to the swells, plastic bottles ride alongside him. As a big wave surfer, Schlebach is determined to conserve his playground. It pains me to see plastic floating through the water, he says. We can no longer turn our backs to the seemingly insurmountable levels of pollution. Schlebachs approach is to revolutionise not just ocean clean-ups, but fashion.

Together with designer Jasper Eales, Schlebach transforms used fabrics such as yacht sails, PVC truck tarps, and display banners into stylish bags and laptop covers. If the materials werent upcycled, they would probably land up in landfills, Schlebach says. Cleaning up the ocean isnt effective if the trash just sits in a dump alongside other items that dont degrade. Schlebach and his team give waste a second life. This is probably as environmentally sustainable as you can get, he says.

With his dedicated team at Sealand Gear, Schlebach has created a chic answer to our plastic problem. No two items are the same, but each one is equally functional and hardy. Every piece is stamped with the name of the person who made it. We are all capable of taking care of our planet one way or another, Schlebach says. When I go surfing, I always know that Im being part of a positive change. Theres a wave of eco-consciousness surging, and its time to join in.

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Surfer cleans his playground by turning pollution into functional fashion - East Coast Radio

A day in the life of a cosmic-ray ‘bookkeeper’ – Symmetry magazine

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When he was growing up, Jonathan LeyVa thought hed follow his passion for race cars and pick a profession in automotive engineering. Instead hes working on what will become one of the worlds most sensitive searches for dark matter, the invisible substance that accounts for more than 85%of the mass of the universe.

LeyVa works in a clean room at the Department of Energys SLAC National Accelerator Laboratory, where crews are building detectors for the latest in a series ofSuper Cryogenic Dark Matter Search, or SuperCDMS,experiments. As an early-career physicist, part of his job is keeping track of how much exposure to cosmic rayshigh-energy particles falling in from spacethe detector components are getting. Researchers want to keep that exposure to a minimum because it could harm their ability to detect dark matter later on.

Ive been interested in cosmology since my senior year in college, LeyVa says, so Im lucky enough to be able to contribute to an exciting project like this at the frontline of dark matter research.

As a freshman at Santa Clara University, LeyVa started out in mechanical engineering, following his childhood dream of doing something with cars. But he soon realized that the field wasnt for him. Inspired by his dad, who holds a physics degree, and by his physics professor at Santa Clara, he began studying physics during his sophomore year.

Having been committed to engineering at first, making this switch was quite daunting, LeyVa says. But he quickly got into the physics world and completed his undergraduate studies in 2017.

Around the time of his graduation, Santa Clara Professor Betty Young suggested that LeyVa spend some time inKent Irwins labat Stanford University, where Young is a visiting scholar. There he learned about SQUIDssuperconducting quantum interference devices used in precision sensors, including those for dark matter searches with SuperCDMS.

This experience got LeyVa hooked on dark matter, whose nature is still unknown and one of the biggest mysteries of modern physics.

He spent the following year with Blas Cabreras team at Stanford, looking for ways of making future SuperCDMS detectorsmore sensitive to lightweight dark matter particles. In 2018he became a member of the SuperCDMS group at SLACs and StanfordsKavli Institute for Particle Astrophysics and Cosmology, which is building detector towers for the current version of the experiment; its scheduled to begin its hunt for dark matter at the Canadian underground lab SNOLAB in the early 2020s.

Nowadays, LeyVa spends a lot of his time in a clean room at SLAC, supporting the SuperCDMS team in assembling the detector towers.

SuperCDMS SNOLAB will initially have four towers, each containing a stack of six silicon and germanium crystals and a bunch of sensitive electronics. Cooled down to almost absolute zero temperature, the crystals will vibrate ever so slightly if a dark matter particle rushes through them, and its these tiny vibrations that the experiment will be looking for.

A major challenge in building the experiment is that the crystals and other detector components are sensitive to particle showers produced when cosmic rays hit the atmosphere. These showers cause unwanted background signals that could make it hard to pick up potential dark matter signals. Thats why the experiment at SNOLAB will operate 6800 feet underground, where its protected from those effects.

It also means that the SuperCDMS team must limit how much detector components are exposed to cosmic rays during the construction of the experiment. Components for the detector towers, for example, are kept three stories underground in a tunnel at Stanford, where they are relatively protected. For the tower assembly and testing, they are brought to SLAC, but each tower can spend only a total of a week at the surface. LeyVa is like a cosmic-ray bookkeeper, closely tracking and logging the number of hours that crystals and hundreds of other detector components are being handled at the lab. Working closely with the teams software developers, hes maintaining and improving the database for that task.

In addition, hes involved in a number of other parts of the project, including noise studies of the system that will collect SuperCDMS data and R&D for future generations of the experiment.

Working on SuperCDMS is just the type of hands-on experience LeyVa enjoys. He loves to experimentat work and in his spare time. It seems that I have too many hobbies for my own good, he says jokingly.

In college, LeyVa volunteered on film crews, which involved him in videography, lighting and acting for several productions. As a media systems technician at his university he set up large-scale sound and video systems for important events. One of the most memorable ones, he says, was a talk by actor Martin Sheen.

Sheen talked about social justice and activism, and I remember his presence created quite a buzz at SCU, he says. I grew up seeing him in some of my favorite movies. He seemed to be a very warm, kind person.

LeyVa is also into photographyan interest that was sparked by his parents. In the early 2000s up to about 2010-ish both my parents ran an advertising agency, taking jobs from some Silicon Valley tech companies. My dad would do photography and my mom would do editing work and graphic design, he says. My parents and my activities in college were influences that may have inspired me to fiddle with instruments in more depth.

In SLACs clean room he continues to find inspiration. SuperCDMS allows him to work on many sides of the projects science and technology, which he considers to be a great learning experience.

Right now, LeyVas planning his next steps in life, such as going to grad school. Particle physics with an emphasis on cosmology and astrophysics is what interests me the most, so becoming involved in cutting-edge cosmological research has been a dream come true for me, he says. Its motivating me to take it to the next level and follow in the footsteps of the great researchers Im working with.

Editor's note: This article is adapted from anarticleoriginally published by SLAC National Accelerator Laboratory.

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A day in the life of a cosmic-ray 'bookkeeper' - Symmetry magazine

Missouri S&T joins dark energy experiment to solve accelerating cosmos mystery – Missouri S&T News and Research

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Missouri S&T has joined the Hobby-Eberly Telescope Dark Energy Experiment (HETDEX) as one of 11 international institutions that are collaborating to define the force causing the accelerated expansion of the universe.

The cosmic acceleration is one of the biggest mysteries in our fundamental physics, says Dr. Shun Saito, cosmologist, assistant professor of physics and leader of Missouri S&Ts HETDEX research group.

To explain this phenomenon we believe began 5 billion years ago, we must introduce an unknown energy component with negative pressure into the universe. That component is what we now call dark energy, Saito says.

Two independent scientific teams unexpectedly discovered cosmic acceleration in 1998. They had initially set out to prove the deceleration of the universe based on a common belief that the universe was dominated by matter, and that its expansion would be slowed down by the pull of gravity. Their discovery that the expansion was not slowing down, but actually accelerating, led to the 2011 Nobel Prize in Physics for three members of those teams.

Saito says the best prevailing model to define dark energy is the cosmological constant Einsteins concept of the energy density or vacuum energy of space he introduced in 1917 when he wantedto stop the universe from collapsing.

HETDEX is stepping into unexplored territory of the universe, says Saito. The discovery potential ofthe experiment is huge we may be able to find that dark energy isnt Einsteins cosmological constant, and this would require a newunderstanding of fundamental physics.

To pursue this quest, Missouri S&T recently joined HETDEX through a memorandum of understanding between the University of Missouri Board of Curators on the recommendation of S&Ts physics department, and the University of Texas at Austin on behalf of its McDonald Observatory, located in the Davis Mountains of West Texas.

The McDonald Observatory contains the Hobby-Eberly Telescope (HET), one of the worlds largest optical telescopes. It has a primary mirror made up of 91 hexagonal segments, and was recently upgraded to a usable aperture of 10 meters with a new wide-field instrument suite, specifically for the HETDEX project.

The immense light-gathering power of this telescope allows us to map out one million distant galaxies that are 9 to 11 billion light-years away, Saito says. Saito will contribute to HETDEX by analyzing the gigantic three-dimensional galaxy maps produced from a set of 78 spectrographs mounted on HET.

Intensity mapping is a novel technique for observing the large-scale structure of the universe its our future, says Saito. It gives us a more efficient and powerful way to extract cosmological information from the data. Even though the technique is mainly used by radio astronomers, we hope to pioneer it in the optical field with HETDEX.

HETDEX observations began in December 2017, and Saito expects the project to continue for about three years.

Over the last year, Missouri S&T has focused on further advancing its astrophysics program.

Saito joined S&T in January from the Max-Planck-Institute for Astrophysics in Germany, where as a postdoctoral researcher, he contributed his cosmological mapping expertise to HETDEX and other spectroscopic, or light-measuring, galaxy surveys. He also contributed to the Baryon Oscillation Spectroscopic Survey in the Sloan Digital Sky Survey-III where space-time measurements were used to investigate dark energy.

Dr. Siddhartha Gurung-Lopez from Centro de Estudios de Fsica del Cosmos de Aragn in Spain recently joined S&Ts HETDEX research group to simulate realistic, computer-generated galaxy populations to compare to the HETDEX observations.

Missouri S&Ts astrophysics program is off to an excellent start, says Dr. Thomas Vojta, chair of the physics department and Curators Distinguished Professor of physics. With our participation in HETDEX and in the LIGO (Laser Interferometer Gravitational-wave Observatory) Scientific Collaboration, we now have cutting-edge research groups in gravitational wave physics and in cosmology. S&T is keeping its eyes wide open to the sky.

HETDEX is a collaboration of The University of Texas at Austin,Pennsylvania State University, Texas A&M University, Ludwig MaximilianUniversity of Munich, Leibniz Institute for Astrophyics Potsdam (AIP), Max PlanckInstitute for Extraterrestrial Physics, Max Planck Institute for Astrophysics, Institutefor Astrophysics in Gttingen, The University of Oxford, The University ofTokyo and Missouri University of Science and Technology. Financial support isprovided by the State of Texas, the United States Air Force, the NationalScience Foundation, partner institutions and the generous contributions of manyprivate foundations and individuals.

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The Secret History of BU’s PRB Observatory and the Telescope It Was Built to House | BU Today – BU Today

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AstronomyHow a discovery at a Hawaii volcano sparked a project with a history unlike anything else on campus

The unassuming observatory dome on the roof of the Physics and Biology Research Building, while unused today, has a storied past. Photo by Jackie Ricciardi

Every day, hundreds of BU students walk down Cummington Mall, unaware that they are passing by a fascinating and unique campus relic, with a story full of drama, mystery, near-disaster, and earth-shattering scientific intrigue.

The unassuming observatory dome on the roof of the Physics and Biology Research Building (PRB) has a storied past: it was built to house a telescope that was eventually used at the South Pole.

The story of this observatory began in 1990 when astronomers at Mauna Kea, a mostly underwater volcano on the island of Hawaii, detected carbon in interstellar clouds that was 10 times brighter than anyone had expected. Interstellar clouds are accumulations of gas, dust, and plasma between star systems in a galaxy. Astronomers can determine the chemical compositions of these clouds by studying the radiation they emanate. The unexpected abundance of carbon detected on Mauna Kea was excitingit could point to something new in the field of astronomy.

The notion was that there may be a lot of atomic carbon that weve never properly mapped, says Thomas Bania, a College of Arts & Sciences professor of astronomy. Carbon is a very important atomic element, and if theres a whole lot of carbon in atomic form that we dont know about, it completely changes the way we study the chemistry of the interstellar medium and the chemistry of molecular clouds, things like that. Detecting that much more carbon would radically change our understanding of the composition of significant portions of the Milky Way Galaxy, transforming our understanding of galactic evolution, the formation of stars and planets, and perhaps even the origin of life in the universe.

Eager to follow this lead, Anthony Stark, an astrophysicist at the Harvard Smithsonian Center for Astrophysics, teamed up with Bania and a team of astronomers from BU and Harvard to create the Antarctic Submillimeter Telescope and Remote Observatory (AST/RO) project. Their group included BU astronomy doctoral students Maohai Huang (GRS00), Alberto Bolatto (GRS01), James Ingalls (GRS98), and undergraduate Edgar Castro (ENG95). Together, they set out to design and build a telescope from scratchone that they would eventually send all the way down to the South Pole for carbon observations.

Observational astronomers run into all sorts of issues when trying to operate a telescope in warmer environments: high wind speeds can shake the instruments, rapid temperature fluctuations cause metal and glass in the telescope to expand and contract, wet weather can damage the sensitive instrumentation, and nosy animals or errant birds can interfere with the telescope.

But these arent issues at the South Pole. It may not be a livable environment, but for over a century, astronomers have been conducting science in this inhospitable place, because with all the inconveniences of moderate climates, scientists have decided the benefits of what they might learn there make getting a bit cold worthwhile.

From 1990 to 1992, the AST/RO telescope was built from scratch at Bell Laboratories in New Jersey and BUs Scientific Instrument Facility in the PRB basement. Its design is called offset Gregorian: once light enters the telescope, it bounces off four mirrors that direct it into a room below, called a Coud room, where it is focused into an image.

As the telescope was being built, a team of BU Facilities Management & Planning workers constructed the test dome on the PRB roof. This was an ideal place to test the telescope since the rooftop had a clear view of the southern horizon (the Life Science & Engineering Building wasnt there yet). One challenge, though, would be getting the telescope up there. The elevator goes only as high as the fifth floorfrom there you have to climb another flight of stairs and then a ladder to get to the porthole that opens onto the roof. The AST/RO researchers made this climb every time they had to get to the observatory. That ladder is still the only way to get to the roof.

There was no way they could bring the telescope in pieces, one by one, up the ladder and through the two-foot-by-two-foot porthole. So the team installed a small crane inside the dome and used it to lift several of the telescope pieces from the fifth floor, through a series of hatchwaysand through a biology labto the observatory. The biggest telescope pieces had to be hoisted onto the roof from a crane outside the building.

Once everything was assembled on the roof, the team spent 18 months testing the instrumentation. Huang remembers spending many long nights at the test site, even taking naps in the control room below the telescope. But every so often, he would wander up to the rooftop dome and enjoy a few quiet moments in the Boston morning. There is nobody there, of course, he says, as the sun is just beginning to crest over the city skyline in the east, and you can see very faryou can see the Charles River, and everything.

In 1995, the AST/RO telescope was finally ready for the South Pole. The team packed it up in a green-painted wooden crate (nicknamed the Green Monster). Its journey to the South Pole took three weeks and multiple modes of transportation: Boston to Los Angeles via truck, Los Angeles to McMurdo Station in Antarctica by ship, then McMurdo to the Amundsen-Scott South Pole Station aboard a C130 military plane.

But it did not go gentle: on the first leg of the trip, the truck carrying it crashed on a highway near Little Rock, Ark. Bania recalls going down to assess the damage with James Jackson, a CAS adjunct professor of astronomy. We had the telescope, all of its computers, all of its electronics, all of the spares, all of the tools that it was going to take to put it back together, all of the documentation for an entire state-of-the-art high-frequency radio observatory in one truck, says Bania. He and Jackson had to decide if it was worth the risk to send the telescope down to the South Pole, or if they should just bring it back to Boston and take stock of the damage there, losing a year in the process.

They decided to send it down to Antarctica. Fortunately, their gamble paid offthe telescope arrived in working condition.

The AST/RO telescope operated at the South Pole from 1995 to 2001. Bania and his students went down every year during the Antarctic summer (winter in the Northern Hemisphere) to perform maintenance and keep it operational.

In the end, though, the project was deemed a flop. After 10 years of work, it turned out that, no, carbon was brighter for a different reasonand so there wasnt this vast reservoir of atomic carbon, Bania says. And proving a negative never leads to a sexy press release.

With that, AST/RO was unceremoniously boxed back up and left in a graveyard of other abandoned scientific instruments at the South Pole. Its now probably buried under ice.

For those who spent hours, days, weeks, and years of their lives on the project, however, it was hardly a flop; all four BU grad students who worked on it got their doctorates and are still doing astronomy research. Huang, for example, is a research professor at the National Astronomical Observatory of China, where he works on the science operation and data processing system for astronomical systems, including the Herschel Space Observatory. What I learned from AST/RO really directly goes into what Im working on now, Huang says. Controlling AST/RO was as difficult as controlling a telescope in space, he says: both are hard to access, so youd better make sure things work the first time. If something breaks, its going to take a very long time to fix.

And, almost 30 years later, the AST/RO test dome remains atop the southwest corner of PRB, a monument to the teams decade of work.

For those researchers like Bania, it stirs up mixed memories. On bad days, I think I wasted 10 years of my life, he says. On good days, I think, well, you know, I had the opportunity to build an observatory from the ground up and operate it. Not many people can say that.

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Astronomers Find a Place With Three Supermassive Black Holes Orbiting Each Other – ScienceAlert

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Astronomers have spotted three supermassive black holes (SMBHs) at the center of three colliding galaxies a billion light years away from Earth. That alone is unusual, but the three black holes are also glowing in x-ray emissions.

This is evidence that all three are also active galactic nuclei (AGN,) gobbling up material and flaring brightly.

This discovery may shed some light on the "final parsec problem," a long-standing issue in astrophysics and black hole mergers.

Astronomers found the three SMBHs in data from multiple telescopes, including the Sloan Digital Sky Survey (SDSS,) the Chandra X-ray Observatory, and the Wide-field Infrared Survey Explorer (WISE.)

The three black holes are wrapped up in an almost unimaginably epic event; a merger of three galaxies. These triplet mergers may play a critical role in how the most massive black holes grow over time.

The astronomers who found it were not expecting to find three black holes in the center of a triple-galaxy merger.

"We were only looking for pairs of black holes at the time, and yet, through our selection technique, we stumbled upon this amazing system," said Ryan Pfeifle of George Mason University in Fairfax, Virginia, the first author of a new paper in The Astrophysical Journal describing these results.

"This is the strongest evidence yet found for such a triple system of actively feeding supermassive black holes."

Triple black hole systems are difficult to spot because there's so much going on in their neighbourhood. They're shrouded in gas and dust that makes it challenging to see into. In this study, it took several telescopes operating in different parts of the electromagnetic spectrum to uncover the three holes. It also took the work of some citizen scientists.

They're not only difficult to spot, but rare.

"Dual and triple black holes are exceedingly rare," said co-author Shobita Satyapal, also of George Mason, "but such systems are actually a natural consequence of galaxy mergers, which we think is how galaxies grow and evolve."

(Hubble/Pfeifle et. al., arXiv, 2019)

The SDSS was the first to spot this triple-merger in visible light, but it was only through Galaxy Zoo, a citizen science project, that it was identified as a system of colliding galaxies.

Then WISE saw that the system was glowing in the infrared, indicating that it was in a phase of galaxy merger when more than one of the black holes was expected to be feeding.

The Sloan and WISE data were just tantalizing clues though, and astronomers turned to the Chandra Observatory and the Large Binocular Telescope (LBT) for more confirmation. Chandra observations showed that there were bright x-ray sources in the center of each galaxy. That's exactly where scientists expect to find SMBHs.

More evidence showing that SMBHs were there arrived from Chandra and NASA's Nuclear Spectroscopic Telescope Array(NuSTAR) satellite. They found evidence of large amounts of gas and dust near one of the black holes.

That's expected when black holes are merging. Other optical light data from the SDSS and the LBT provided spectral evidence that's characteristic of the three SMBHs feeding.

(NASA/CXC/NGST)

"Optical spectra contain a wealth of information about a galaxy," said co-author Christina Manzano-King of University of California, Riverside. "They are commonly used to identify actively accreting supermassive black holes and can reflect the impact they have on the galaxies they inhabit."

With this work, the team of astronomers have developed a way to find more of these triple black hole systems.

"Through the use of these major observatories, we have identified a new way of identifying triple supermassive black holes. Each telescope gives us a different clue about what's going on in these systems," said Pfeifle. "We hope to extend our work to find more triples using the same technique."

They may have also shed some light on the final parsec problem.

The final parsec problem is central to our understanding of binary black hole mergers. It's a theoretical problem that says when two black holes approach each other, their excessive orbital energy stops them from merging. They can get to within a few light years, then the merging process stalls.

When two black holes initially approach each other, their hyperbolic trajectories carry them right past each other. Over time, as the two holes interact with stars in their vicinity, they slingshot the stars gravitationally, transferring some of their orbital energy to a star each time they do it. The emission of gravitational waves also decreases the black holes' energy.

Eventually the two black holes shed enough orbital energy to slow down and approach each other more closely, and come to within just a few parsecs of each other.

The problem is, as they close the distance, more and more matter is ejected from their vicinity via sling-shotting. That means there's no more matter for the black holes to interact with and shed more orbital energy. At that point, the merging process stalls. Or it should.

Yet astrophysicists know that black holes merge because they've witnessed the powerful gravitational waves. In fact, LIGO (Laser Interferometry Gravitational-Wave Observatory) is discovering a black hole merger about once a week. How they merge with each other at the end is called the final parsec problem.

The team behind this study thinks that they might have an answer. They think that a third black hole, like they've observed in this system, could provide the boost needed to get two holes to merge.

As a pair of black holes in a trinary system approach each other, the third hole could influence them to close the final parsec and merge.

According to computer simulations, about 16% of pairs of supermassive black holes in colliding galaxies will have interacted with a third supermassive black hole before they merge.

Those mergers would produce gravitational waves, but the problem is that those waves would be too low-frequency for LIGO or the VIRGO observatory to detect.

(ESA/NASA/LISA)

To detect those, scientists may have to rely on future observatories like LISA, ESA/NASA's Laser Interferometer Space Antenna. LISA will observe lower frequency gravitational waves than LIGO or VIRGO and is better-equipped to find super-massive black holes merging.

The paper presenting these results is titled "A Triple AGN in a Mid-Infrared Selected Late Stage Galaxy Merger."

This article was originally published by Universe Today. Read the original article.

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Astronomers Find a Place With Three Supermassive Black Holes Orbiting Each Other - ScienceAlert

Sounds of space: Janna Levin kicks off celebration of the 20th anniversary of the Commonwealth Honors College – The Massachusetts Daily Collegian

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Renowned cosmologist speaks about major scientific discovery in her latest book

By Sara Abdelouahed, Collegian CorrespondentOctober 1, 2019

On Thursday, Sept. 26th, the Commonwealth Honors College began a year-long series of events celebrating its 20th anniversary at the University of Massachusetts. This preliminary event, the Williamson Lecture, featured world-renowned author and cosmologist Janna Levin.

The focus of Levins talk was her newest book, Black Hole Blues and Other Songs from Outer Space, which explores the discovery of gravitational waves produced by black holes. Her knowledge and connections to those involved in the discovery allowed her to bring the story of the 50-year endeavor to life.

Honors College Dean Gretchen Gerzina introduced Levin with a brief summary of her background and experience. Levin is well known in the astronomy community for her work in understanding black holes and the cosmology of extra dimensions. She previously presented the NOVA show Black Hole Apocalypse on PBS, the shows first female presenter in 35 years. Along with her newest release, Levin has now published three books.

According to Gerzina, she is the rock star of astrophysics.

Her presence in the scientific community is known even to college students, including freshman physics major Garrett Mann. Mann had previously seen Levin on a YouTube show with astrophysicist Neil deGrasse Tyson. Manns curiosity and desire to know how [blackholes] work drew him to the event to hear her speak in person.

Other students echoed his desire to learn more. Freshman biochemistry major Aurora OConnor came without prior knowledge of the subject matter.

I know nothing about black holes, OConnor said. Maybe I should learn something about them.

Levin highlighted the exciting ups and downs of the scientific breakthrough using metaphors, anecdotes, and humor, allowing listeners of all backgrounds to comprehend the significance of the feat.

When asked about her favorite aspect of talking to college students, Levin said that everyone was really engaged. Its so fun when people are that present and curious and energized.

Students were eager to ask questions after the discussion, quickly lining up for the opportunity to talk to Levin and have their copies of her book signed. Amherst Books sold Levins newest publication at the event.

Colby Pratt, a freshman astronomy major at the University, said that her personal interest drew her to the event. Not only did she want to better understand black holes, but she [wanted] to be able to share that with other people.

Levin shares a similar philosophy.

[Physics] is like a gift, it belongs to you, she explained. Its something you inherit as soon as you learn it and then its yours to teach to somebody else. She said that the ability to share learned information with others was something that [she] found just incredibly, overwhelmingly exciting.

The 20th Anniversary celebration of the Commonwealth Honors College continues on Oct. 1 with a lecture from Banu Subramaniam, titled A Scientific Renaissance: Gender, Race, and the Practice of Science.

Sara Abdelouahed can be reached at [emailprotected].

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Sounds of space: Janna Levin kicks off celebration of the 20th anniversary of the Commonwealth Honors College - The Massachusetts Daily Collegian