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News Article | May 19, 2017
Site: www.prweb.com

Beginning June 1st, 2017, The Whistler Group, Inc., a leading industry supplier of radio scanners and mobile electronics, will offer a feature upgrade program for the Pro-18, Pro-668 and PSR-800 scanners. For the cost of $59.99, customers will be able to send in their scanner to Whistler where the unit will be upgraded to receive DMR and MotoTRBO™. The cost of shipping the unit to Whistler will be covered by the customer and Whistler will cover the cost of shipping the unit back to the customer when the upgrade is complete. Upgrades are expected to take up to 2 weeks and all units must be in working condition with no visible damage to be eligible for this upgrade program. By upgrading their scanner, customers will now receive DMR making their scanner capable of monitoring the following unencrypted channels/systems which were previously not available for legacy scanners: To learn more visit the Whistler Group website at http://www.whistlergroup.com MotoTRBO™ is a registered trademark of Motorola, Inc.


News Article | May 24, 2017
Site: www.businesswire.com

CHICAGO--(BUSINESS WIRE)--Littelfuse, Inc., the global leader in circuit protection, expands its award-winning line of high-speed fuses for protection of power semiconductor devices used in industrial equipment. The POWR-SPEED® PSR Series line of square body fuses combine extreme current limitation, excellent cycling capability, and low watt-loss design in a direct bus-bar mount construction with blown fuse indication included. The square body design accommodates high current ratings with enhanced DC performance in a smaller footprint than the traditional round body style of high-speed fuses. “DC applications such as energy storage, electric vehicle charging stations, and uninterruptible power supplies (UPS) are increasing and the manufacturers of these products have a growing need for better DC protection of their system components,” explains Craig Greeson, product manager, power fuses at Littelfuse. “The POWR-SPEED® PSR Series has the best DC protection performance available in the market today.” Key benefits of the new PSR Series include: The expanded line includes a voltage range of 550VAC to 1300VAC, 500VDC to 1000VDC. Current ratings range from 125 to 2000A and interrupt ratings are 200kA AC and 150kA DC. These fuses are used in many applications including power conversion equipment, low and medium voltage drives, DC common bus protection, battery protection, power supplies, and industrial heaters. Technical data and application support are available on-line at www.littelfuse.com/powerspeed or by calling our technical support line at 1-800-832-3873. Design engineers looking for a customized fuse design are encouraged to contact the Littelfuse Application Support Team. Founded in 1927, Littelfuse is the world leader in circuit protection with growing global platforms in power controls and sensing. The company serves customers in the electronics, automotive and industrial markets with technologies including fuses, semiconductors, polymers, ceramics, relays and sensors. Littelfuse has over 10,000 employees in more than 40 locations throughout the Americas, Europe and Asia. For more information, please visit the Littelfuse website: littelfuse.com.


News Article | April 19, 2017
Site: www.rdmag.com

Researchers at UC Riverside's Akbari lab have brought a new strain of red-eyed mutant wasps into the world. The wasps were created to prove that CRISPR gene-slicing technology can be used successfully on the tiny parasitic jewel wasps, giving scientists a new way to study some of the wasp's interesting biology, such as how males can convert all their progeny into males by using selfish genetic elements. No one knows how that selfish genetic element in some male wasps "can somehow kill the female embryos and create only males," said Omar Akbari, an assistant professor of entomology who led the research team. "To understand that, we need to pursue their PSR (paternal sex ratio) chromosomes, perhaps by mutating regions of the PSR chromosome to determine which genes are essential for its functionality." Enter the relatively new CRISPR technology, which allows scientists to inject components like RNA and proteins into an organism with instructions to find, cut and mutate a specific piece of DNA. Then researchers can see how disrupting that DNA affects the organism. The end goal, in Akbari's case, is to better understand the biology of wasps and other insects, so they can find a way to control insects that destroy crops or spread diseases like malaria. But the first step is figuring out how to use the CRISPR technology in such a small organism, something no one had ever done before, in large part because the work is pretty daunting, Akbari said. This is because jewel wasps lay their tiny eggs inside a blowfly pupa, which had to be peeled back to expose the teensy eggs. How tiny? Imagine the blowfly egg sac as about the size of a small bean, Akbari said, and Jewel wasp eggs "about a quarter the size of a grain of rice....You're essentially pulling a small egg out of a larger egg, injecting it with components to mutate the DNA and then putting it back into the bigger egg to develop." In the case of Akbari's mutant wasps, the team decided to slice the genes that control the color of the wasp's normally black eyes. "We wanted to target a gene that would be obvious, and we knew from previous studies that if the gene for eye pigmentation was knocked out, they would have red eyes, so this seemed like a good target for gene disruption," Akbari said. "Big beautiful red eyes are something you won't miss." But creating that disruption took some doing--well, a lot of doing, Akbari said. "You have to use a very-very fine needle and a microscope and individually inject hundred to thousands of embryos, but in the end, we developed a protocol that can be used to cut the DNA in this organism and we showed that it works." The technique is challenging, Akbari said, "but it is learnable. You need a really steady hand and it requires a lot of patience in micro manipulation that one can learn over time. Ming Li, a postdoctoral researcher in our lab has mastered the technique." And those scarlet-orbed wasps? They won't be going away anytime soon. The cuts in the DNA created a mutant wasp with heritable traits, which means those red eyes will be passed down to all their offspring in the future - an important quality for researchers who are looking for a stable line of insects to study.


News Article | April 19, 2017
Site: www.eurekalert.org

IMAGE:  The red-eyed mutant Jewel wasp, seen on the right, was created in the lab using CRISPR technology. An unmodified Jewel wasp is seen on the left. view more RIVERSIDE, Calif. (http://www. )-- Researchers at UC Riverside's Akbari lab have brought a new strain of red-eyed mutant wasps into the world. The wasps were created to prove that CRISPR gene-slicing technology can be used successfully on the tiny parasitic jewel wasps, giving scientists a new way to study some of the wasp's interesting biology, such as how males can convert all their progeny into males by using selfish genetic elements. No one knows how that selfish genetic element in some male wasps "can somehow kill the female embryos and create only males," said Omar Akbari, an assistant professor of entomology who led the research team. "To understand that, we need to pursue their PSR (paternal sex ratio) chromosomes, perhaps by mutating regions of the PSR chromosome to determine which genes are essential for its functionality." Enter the relatively new CRISPR technology, which allows scientists to inject components like RNA and proteins into an organism with instructions to find, cut and mutate a specific piece of DNA. Then researchers can see how disrupting that DNA affects the organism. The end goal, in Akbari's case, is to better understand the biology of wasps and other insects, so they can find a way to control insects that destroy crops or spread diseases like malaria. But the first step is figuring out how to use the CRISPR technology in such a small organism, something no one had ever done before, in large part because the work is pretty daunting, Akbari said. This is because jewel wasps lay their tiny eggs inside a blowfly pupa, which had to be peeled back to expose the teensy eggs. How tiny? Imagine the blowfly egg sac as about the size of a small bean, Akbari said, and Jewel wasp eggs "about a quarter the size of a grain of rice....You're essentially pulling a small egg out of a larger egg, injecting it with components to mutate the DNA and then putting it back into the bigger egg to develop." In the case of Akbari's mutant wasps, the team decided to slice the genes that control the color of the wasp's normally black eyes. "We wanted to target a gene that would be obvious, and we knew from previous studies that if the gene for eye pigmentation was knocked out, they would have red eyes, so this seemed like a good target for gene disruption," Akbari said. "Big beautiful red eyes are something you won't miss." But creating that disruption took some doing--well, a lot of doing, Akbari said. "You have to use a very-very fine needle and a microscope and individually inject hundred to thousands of embryos, but in the end, we developed a protocol that can be used to cut the DNA in this organism and we showed that it works." The technique is challenging, Akbari said, "but it is learnable. You need a really steady hand and it requires a lot of patience in micro manipulation that one can learn over time. Ming Li, a postdoctoral researcher in our lab has mastered the technique." And those scarlet-orbed wasps? They won't be going away anytime soon. The cuts in the DNA created a mutant wasp with heritable traits, which means those red eyes will be passed down to all their offspring in the future - an important quality for researchers who are looking for a stable line of insects to study. The results were just published in Nature's Scientific Reports in an article called "Generation of heritable germline mutations in the jewel wasp Nasonia vitripennis using CRISPR/Cas9." Besides Akbari, the authors include Li, Abigail Chong and Bradley J. White of UCR and Lauren Yun Cook Au, Deema Douglah and Patrick M. Ferree from Claremont McKenna College in Claremont. The research was supported by UCR startup funds.


News Article | April 19, 2017
Site: www.rdmag.com

Researchers at UC Riverside's Akbari lab have brought a new strain of red-eyed mutant wasps into the world. The wasps were created to prove that CRISPR gene-slicing technology can be used successfully on the tiny parasitic jewel wasps, giving scientists a new way to study some of the wasp's interesting biology, such as how males can convert all their progeny into males by using selfish genetic elements. No one knows how that selfish genetic element in some male wasps "can somehow kill the female embryos and create only males," said Omar Akbari, an assistant professor of entomology who led the research team. "To understand that, we need to pursue their PSR (paternal sex ratio) chromosomes, perhaps by mutating regions of the PSR chromosome to determine which genes are essential for its functionality." Enter the relatively new CRISPR technology, which allows scientists to inject components like RNA and proteins into an organism with instructions to find, cut and mutate a specific piece of DNA. Then researchers can see how disrupting that DNA affects the organism. The end goal, in Akbari's case, is to better understand the biology of wasps and other insects, so they can find a way to control insects that destroy crops or spread diseases like malaria. But the first step is figuring out how to use the CRISPR technology in such a small organism, something no one had ever done before, in large part because the work is pretty daunting, Akbari said. This is because jewel wasps lay their tiny eggs inside a blowfly pupa, which had to be peeled back to expose the teensy eggs. How tiny? Imagine the blowfly egg sac as about the size of a small bean, Akbari said, and Jewel wasp eggs "about a quarter the size of a grain of rice....You're essentially pulling a small egg out of a larger egg, injecting it with components to mutate the DNA and then putting it back into the bigger egg to develop." In the case of Akbari's mutant wasps, the team decided to slice the genes that control the color of the wasp's normally black eyes. "We wanted to target a gene that would be obvious, and we knew from previous studies that if the gene for eye pigmentation was knocked out, they would have red eyes, so this seemed like a good target for gene disruption," Akbari said. "Big beautiful red eyes are something you won't miss." But creating that disruption took some doing--well, a lot of doing, Akbari said. "You have to use a very-very fine needle and a microscope and individually inject hundred to thousands of embryos, but in the end, we developed a protocol that can be used to cut the DNA in this organism and we showed that it works." The technique is challenging, Akbari said, "but it is learnable. You need a really steady hand and it requires a lot of patience in micro manipulation that one can learn over time. Ming Li, a postdoctoral researcher in our lab has mastered the technique." And those scarlet-orbed wasps? They won't be going away anytime soon. The cuts in the DNA created a mutant wasp with heritable traits, which means those red eyes will be passed down to all their offspring in the future - an important quality for researchers who are looking for a stable line of insects to study.


News Article | April 19, 2017
Site: www.chromatographytechniques.com

Researchers at UC Riverside's Akbari lab have brought a new strain of red-eyed mutant wasps into the world. The wasps were created to prove that CRISPR gene-slicing technology can be used successfully on the tiny parasitic jewel wasps, giving scientists a new way to study some of the wasp's interesting biology, such as how males can convert all their progeny into males by using selfish genetic elements. No one knows how that selfish genetic element in some male wasps "can somehow kill the female embryos and create only males," said Omar Akbari, an assistant professor of entomology who led the research team. "To understand that, we need to pursue their PSR (paternal sex ratio) chromosomes, perhaps by mutating regions of the PSR chromosome to determine which genes are essential for its functionality." Enter the relatively new CRISPR technology, which allows scientists to inject components like RNA and proteins into an organism with instructions to find, cut and mutate a specific piece of DNA. Then researchers can see how disrupting that DNA affects the organism. The end goal, in Akbari's case, is to better understand the biology of wasps and other insects, so they can find a way to control insects that destroy crops or spread diseases like malaria. But the first step is figuring out how to use the CRISPR technology in such a small organism, something no one had ever done before, in large part because the work is pretty daunting, Akbari said. This is because jewel wasps lay their tiny eggs inside a blowfly pupa, which had to be peeled back to expose the teensy eggs. How tiny? Imagine the blowfly egg sac as about the size of a small bean, Akbari said, and Jewel wasp eggs "about a quarter the size of a grain of rice....You're essentially pulling a small egg out of a larger egg, injecting it with components to mutate the DNA and then putting it back into the bigger egg to develop." In the case of Akbari's mutant wasps, the team decided to slice the genes that control the color of the wasp's normally black eyes. "We wanted to target a gene that would be obvious, and we knew from previous studies that if the gene for eye pigmentation was knocked out, they would have red eyes, so this seemed like a good target for gene disruption," Akbari said. "Big beautiful red eyes are something you won't miss." But creating that disruption took some doing--well, a lot of doing, Akbari said. "You have to use a very-very fine needle and a microscope and individually inject hundred to thousands of embryos, but in the end, we developed a protocol that can be used to cut the DNA in this organism and we showed that it works." The technique is challenging, Akbari said, "but it is learnable. You need a really steady hand and it requires a lot of patience in micro manipulation that one can learn over time. Ming Li, a postdoctoral researcher in our lab has mastered the technique." And those scarlet-orbed wasps? They won't be going away anytime soon. The cuts in the DNA created a mutant wasp with heritable traits, which means those red eyes will be passed down to all their offspring in the future - an important quality for researchers who are looking for a stable line of insects to study.


News Article | February 16, 2017
Site: www.businesswire.com

NEW YORK--(BUSINESS WIRE)--Mantle Ridge LP (“Mantle Ridge”), an investment firm that owns approximately 4.9% of the outstanding common shares of CSX Corp. (NASDAQ:CSX) (the “Company”), today sent the following letter to the CSX Board of Directors. I read with interest your press release announcing a special meeting to poll shareholders on their views about our proposals. Hunter and I are committed exclusively to finding ways to maximize shareholder value here. We know that the Board shares this objective. Because every day’s delay in beginning to implement Precision Scheduled Railroading (PSR) under conditions that enable its success costs the Company dearly, it is in the shareholders’ best interest that we promptly resolve this through negotiation rather than wait months. Continuing the exploration for common ground makes sense. As I have said in my earliest communications with you, and in each correspondence since, I am convinced there is ample room for agreement. We have been standing by since last week for constructive counterproposals and we are ready to discuss them as they arrive. Early on in this process, I explained that we were looking to you to help us shape a solution that the Board would embrace, and that would best advance the shareholders’ objective of a swift and certain transformation. I made that clear in my January 24th letter to the Board. The relevant passage is below: “You Are Necessarily the Leader of this Process For a variety of reasons, you are uniquely positioned to lead: you know the Company, the people, and the dynamics at the Board and in the executive ranks; you have had dealings with me, and you know Hunter by reputation; you have engineered countless deals; and you have been through a somewhat similar (but the differences are greater than the similarities) experience in the past. We seek your guidance in supporting your leadership.” We remain hopeful, Ned, that you and we can together craft a solution that works for all. If so the shareholder wins big. In the spirit of continuing to engage with you pending the special meeting, I thought it would be useful for me to give you some high-level input on what you said in the press release, to discuss process, and a proposed path forward. The press release suggests that Mantle Ridge is looking for “substantial board representation.” In fact, we are requesting only a single Mantle Ridge representative on the Board (me). Neither Hunter nor the other Board slots we discussed represents Mantle Ridge in any way. The press release goes on to say that Mantle Ridge wants to designate six directors and suggests that granting our ask would grant us effective control. In fact, you and I were engaged in a process to identify high quality independent people that the Board and Mantle Ridge could embrace. As part of that process, I gave you a list (including lengthy bios) of 11 exceptional individuals with broad and relevant experience. None of them has any relationship with me outside of my efforts to recharge the CSX Board. Each one is entirely independent of me and is committed to acting in the best interests of all of the shareholders. Each one would make a superb board member. You indicated to me that you were impressed with the list. You agreed that the Board would not consider any of the people on the list surrogates of mine, or anything but truly independent and qualified. I offered to arrange for you and the governance committee to meet any or all of the candidates. You assured me that there was no need to do that, since you were confident that together we could settle on people from that list once we settled on the other issues. Why are we asking that new directors be added? As we’ve discussed, Precision Scheduled Railroading requires dramatic operational and cultural change. Change like that starts at the top, with significant new blood on the Board not wed to the old ways or legacy decisions and with no ties to any previous strategy or any one. The messaging to all concerned constituencies that the external change agent – Hunter – is coming in with very substantial support empowers Hunter. Conversely, without enormous board support, the outcome and rate of the transformation will be at risk. As Hunter said in our own press release, “if we create the right conditions for success, we have the best chances for success.” When I mentioned “conditions for success,” you asked what that means. I explained to you and the rest of the Board that the conditions at CP were ideal (from early on during the transformation, new independent directors comprised a majority of the board). But I also said I think that something less than that would be workable at CSX and that I was open to exploring any proposals that you suggested could work. More generally, I have said repeatedly, that my view of how well a proposed governance deal will affect Hunter’s ability to succeed is based on an assessment of roll-offs, additions, and board roles/committees. No one of these dimensions can be considered on its own. They are a package. It is a balancing act. And different packages and permutations can work. The question before us is which package or permutation is acceptable to the Board, and sufficiently empowers Hunter to deliver for the shareholders. I must confess, however, I was made somewhat cautious during this process. The Board’s letter to Hunter proposed that he be hired for only two years, not the four we explained was necessary. This raised concerns relating to the Board’s commitment to the transformation. That lack of commitment could create material risk to the timely and successful implementation of PSR. Having lived this, I cannot overemphasize the risk this brings to our delivering to shareholders the swift and certain transformation they want and expect. With that in mind, I told you that the addition of three independent directors the Board offered was better than its initial offer of two. I suggested you take four back for consideration. I also made clear that I was open to different approaches concerning how quickly the Board size comes down. I note that Tuesday’s press release could be read to say we were looking for the Board to drop down to 12 after John Breaux ages out in 2018. I never made that ask, and in fact I had just assumed that the reconstituted Board would find a replacement when he chooses to step down. I said that I considered Mr. Breaux a valuable member of the Board and explicitly said more than once that a retirement age cap should not get in the way of his continuing to serve for many years if that was his preference. I continue to hope he stays for a while. Similarly, when we discussed committee composition and leadership, I gave you my thoughts but made clear that when the time comes we would sit down and agree on a list. I said that it made sense for the new directors to occupy a minority of the committee chairs. I explained that the committees that were most important for the transformation to succeed were Governance and Compensation and that it made sense that two of the independents occupy those. For avoidance of doubt, Mantle Ridge has been seeking to add to the Board only one representative, plus Hunter, plus independent people. The sole prism through which we should evaluate the number of new directors that should be added to the Board is the best interests of the Company. That the board restructuring we are discussing comes in connection with an activist whose holdings are 5% of the Company, or 1%, or 15% is irrelevant. The size and nature of the restructuring should be solely a function of the shareholders’ best interests, which I think you agree requires installing Hunter and creating the conditions for him to succeed. No other consideration – the size of the proposing shareholder or otherwise – should be a factor. Mantle Ridge is in no way seeking or gaining control, and remains open to discussions about the details. This is not a “battle for control”. The press release suggests that Hunter is asking for a $300 million compensation package. In our view, that’s a major mischaracterization. Regrettably, it has confused the shareholders. Hunter’s compensation package adds up to approximately $32 million per year for four years, of which approximately $20 million per year is explicitly performance based and should therefore be discounted in the customary and substantial way performance based grants normally get discounted for compensation and accounting purposes. In addition to Hunter’s compensation package, there is a one-time cost of extraction from CP (i.e., the value Hunter forfeited in order to free him up from his two-year non-compete and allow him to work at CSX), which should not be viewed as compensation. That cost is $84 million plus a gross up of somewhere between $0 and $23 million (depending on Hunter’s tax position). Importantly, as you know, in no way does Mantle Ridge receive any economic gain from this arrangement. As a matter of fact, Mantle Ridge’s investors assumed this liability to effect the release of Mr. Harrison from his non-compete and make him available to solve CSX’s succession problem. This just covers that cost. Aside from our different ways of characterizing the one-time cost of extraction, the main difference between our number and yours is the value of the option grant. The $160 million referenced in the Company’s press release confuses the matter. That figure does not fairly represent the economic cost of this package to the Company, or its value to Hunter. And it should not be the basis for evaluating the package. As you and I discussed, in our view the proper way to think about that value is to view it against the unaffected stock price of $36.88 rather than today’s “Hunter rally” price. The market has already baked in anticipated strong execution by Hunter worth $10 per share and, since his strike price will be based on the inflated date-of-grant price, Hunter will receive no in-the-money value for that first $10 per share of his value creation. For this reason, we believe the economically correct view is that he was granted an option that is $10 per share out of the money on the day of grant. Viewed that way, the Black-Scholes value of the grant is only $78 million, and half of that grant is performance based (based on goals that no other CEO in this industry has reached) and as such should be accounted for at a large discount. It only has meaningful value if Hunter knocks the cover off the ball, in which case the magnitude of the package would be de minimis relative to the value he uniquely could create. And it would certainly be deserved. This is the correct characterization of the economic cost of the package to shareholders and the economic value to Hunter. We had discussed this verbally a couple of times and you seemed to follow the analysis and accept the conclusion. Importantly, because under the proposal Hunter’s options are so far out of the money relative to the pre-Hunter price, it is important that we never lose sight of the fact that his package is worth very little unless he performs spectacularly. As with the governance proposal, we are looking for your guidance in shaping a deal that works for all. Ned, a lot of the points you raised in the press release could easily have been addressed through continued back and forth, but the back and forth stopped. Hunter and I (and our counsel) repeatedly asked for a counter on the compensation and governance terms, but we never received one. Instead, we have a special meeting that will distract current management and – more importantly – will delay roll out of PSR and disrupt the Company’s operations. When you compare this cost with the cost of the questions at issue, it becomes clear that the Board should redouble its effort to get a settlement. Again, you are clearly the best party to get us there, and we continue to look to you for guidance on proposals that would work. With that in mind, as you and I gear up for the special meeting, I remain open to engaging constructively with you in a way that could bring this to a more rapid and satisfactory close for the shareholders. Over the last 24 hours, I have spoken to many shareholders of the Company. Without exception, they share our eagerness to get Hunter into the seat as quickly as possible under conditions for success that would enable him replicate the CP transformation. They agree that the cost to the Company of delaying this outcome until a Special Meeting gets scheduled in April, or possibly later, is dear. None of us wants to wait. I would ask the Board to consider the clarification of the underlying economics of Hunter’s compensation package we provide above and to reconsider whether they can consider accepting it, or at least providing a counter proposal. Concerning the governance proposal, this is a soft and tricky exercise in judgment that balances additions, roll-offs, and roles/committees. If the Board can commit to a process that promptly gets us to a four-year deal with Hunter, that will in my mind be a sufficient gesture of commitment that we can go forward with the addition of me and Hunter and three of the independent directors (rather than the four in our last proposal), with continued discussion concerning roll-offs, committee composition, and roles. Ned, we have come a long way. We are close. We owe it to the shareholders to get a deal done promptly. Let’s do it. If you are willing, we are glad to meet in person and hammer this out this weekend, hopefully delivering good news to the shareholders early next week. As before, I've taken the liberty of copying the other members of the Board as well as Ms. Fitzsimmons. Ms. Fitzsimmons, please pass on copies of this letter to every other Board member. Very truly yours, Mantle Ridge is a private investment firm founded by Paul Hilal. Mantle Ridge seeks to help create enduring value, and believes that constructive and cooperative engagement between boards, management teams, and engaged shareholders is the best means to achieving that goal. Mantle Ridge’s approach is informed by extensive research and a depth of experience in value investing, activist engagements, corporate governance, and business operations, in the context of diverse sector expertise. For more information, go to www.mantleridge.com. This press release contains forward-looking statements within the meaning of Section 27A of the Securities Act of 1933, as amended, and Section 21E of the Securities Exchange Act of 1934 (as amended, the “Exchange Act”), regarding, among other things, Mantle Ridge’s plans to distribute a definitive proxy statement, as well as the benefits of Precision Scheduled Railroading (and the cost of delaying its implementation). Such forward-looking statements involve known and unknown risks, uncertainties and other factors that may cause actual results, performance or achievements to be materially different from any future results, performance or achievements expressed or implied by such forward-looking statements. Readers are cautioned not to place undue reliance on these forward-looking statements, which speak only as of the date thereof. Mantle Ridge undertakes no obligation to publicly release the result of any revisions to these forward-looking statements that may be made to reflect events or circumstances after the date hereof or to reflect the occurrence of unanticipated events. Mantle Ridge LP (“Mantle Ridge”) and certain of its affiliates intend to file with the Securities and Exchange Commission (the “SEC”) a proxy statement and accompanying proxy card to be used to solicit proxies in connection with the upcoming special meeting of shareholders (the “Special Meeting”) of CSX Corporation (the “Company”) announced on February 14, 2017 and the election of a slate of director nominees at the 2017 annual shareholders meeting of the Company (the “Annual Meeting”). The participants in the solicitation are anticipated to include MR Argent Offshore AB Ltd., MR Argent Offshore BB Ltd., MR Argent Offshore CB 01 Ltd., MR Argent Offshore CB 02 Ltd., MR Argent Offshore CB 03 Ltd., MR Argent Offshore CB 04 Ltd., MR Argent Offshore CB 05 Ltd., MR Argent Offshore CB 07 Ltd. (collectively, the “MR Funds”), MR Argent Fund CE LP, MR Argent Advisor LLC (the “Advisor”), Mantle Ridge GP LLC, Mantle Ridge LP, MR Argent GP LLC, MR S and P Index Annual Reports LLC (“Shareholder”), Mr. Hilal (collectively with the MR Funds, MR Argent Fund CE LP, the Advisor, Mantle Ridge GP LLC, Mantle Ridge LP, MR Argent GP LLC and Shareholder, the “Mantle Ridge Parties”) and E. Hunter Harrison. MANTLE RIDGE STRONGLY ADVISES ALL SHAREHOLDERS OF THE COMPANY TO READ THE DEFINITIVE PROXY STATEMENTS AND OTHER DOCUMENTATION RELATED TO THE SOLICITATION OF PROXIES BY MANTLE RIDGE AND ITS AFFILIATES FROM SHAREHOLDERS OF THE COMPANY FOR USE AT THE SPECIAL MEETING AND ANNUAL MEETING WHEN THEY BECOME AVAILABLE, BECAUSE THEY WILL CONTAIN IMPORTANT INFORMATION, INCLUDING INFORMATION RELATING TO THE PARTICIPANTS IN SUCH PROXY SOLICITATION. SUCH DEFINITIVE PROXY STATEMENTS, OTHER PROXY MATERIALS AND ANY OTHER RELEVANT DOCUMENTATION WILL BE AVAILABLE AT NO CHARGE ON THE SEC’S WEB SITE AT HTTP://WWW.SEC.GOV. IN ADDITION, MANTLE RIDGE WILL PROVIDE COPIES OF THE DEFINITIVE PROXY STATEMENT AND OTHER MATERIALS WITHOUT CHARGE, WHEN AVAILABLE, UPON REQUEST. REQUESTS FOR COPIES SHOULD BE DIRECTED TO D.F. KING & CO., INC., 48 WALL STREET, 22ND FLOOR, NEW YORK, NEW YORK 10005 (CALL COLLECT: (212) 269-5550) OR EMAIL: INFO@DFKING.COM. As of the date hereof, the Mantle Ridge Parties beneficially own (within the meaning of Rule 13d-3 under the Exchange Act) an aggregate of approximately 44,352,597 shares of common stock, $1.00 par value, of the Company (the “Shares”). Of those approximately 44,352,597 Shares, MR S AND P Index Annual Reports LLC owns approximately 6 Shares in record name. As of the date hereof, MR Argent Fund CE LP possesses economic exposure to an aggregate of 570,600 Shares due to certain cash-settled total return swap agreements. MR Argent Fund CE LP also holds American-style call options to purchase cash-settled total return swaps referencing an aggregate of 105,232 Shares. Mantle Ridge LP, as the sole member of the Advisor, which is in turn the advisor to the MR Funds, may be deemed to have the shared power to vote or direct the vote of (and the shared power to dispose or direct the disposition of) the approximately 44,352,590 Shares held for the accounts of the MR Funds and the approximately 6 Shares directly owned by Shareholder and, therefore, may be deemed to be the beneficial owner of such Shares. MR Argent Advisor LLC, as the advisor to, and holder of 100% of the noneconomic voting interests in, the MR Funds, may be deemed to have the shared power to vote or direct the vote of (and the shared power to dispose or direct the disposition of) the approximately 44,352,590 Shares held for the accounts of the MR Funds and, therefore, may be deemed to be the beneficial owner of such Shares. By virtue of his position as the managing member of Mantle Ridge GP LLC, the general partner of Mantle Ridge LP, which is in turn the sole member of both Shareholder and MR Argent Advisor LLC, the advisor to the MR Funds, Mr. Hilal may be deemed to have the shared power to vote or direct the vote of (and the shared power to dispose or direct the disposition of) of the approximately 44,352,590 Shares held for the accounts of the MR Funds and the approximately 6 Shares directly owned by Shareholder and, therefore, may be deemed to be the beneficial owner of such Shares.


News Article | February 23, 2017
Site: globenewswire.com

NEWS RELEASE   REGULATED INFORMATION  INSIDE INFORMATION    Record revenue of $30 million, up 70% from 2015 Total test volume up 57% over 2015 Conference call for analysts and investors today at 15:00 CET / 09:00 EST, details below IRVINE, CA, and HERSTAL, BELGIUM - 7:00 AM, February 23, 2017 - MDxHealth SA (Euronext: MDXH.BR), or the "Company", today announced its financial results for the financial year ended December 31, 2016. Dr. Jan Groen, Chief Executive Officer of MDxHealth commented: "The strong progress made in 2016 with the accelerated adoption of ConfirmMDx and the successful launch of our first liquid biopsy test, SelectMDx, demonstrates the significant potential of MDxHealth's world leading urological oncology franchise." "With the ongoing roll out of SelectMDx, together with the planned launch of our second liquid biopsy test AssureMDx for Bladder Cancer this year, the Company continues to make rapid and meaningful progress in the expansion of its portfolio of products that address unmet medical needs for cancer patients, while leveraging the commercial infrastructure we have built over the last several years.  We are pleased with our progress in 2016 and believe MDxHealth is well positioned to build strong, sustainable growth in 2017 and beyond." Strong growth of ConfirmMDx driven by wide market adoption and continued expansion of reimbursement coverage NCCN Guideline inclusion and increasing adoption has enabled MDxHealth to secure 19 reimbursement contracts, while 28 payors issued positive medical coverage policies. The test is now also covered under the California Medical Assistance Program (Medi-Cal), the single largest state-run public health program in the US with nearly 12 million enrollees. This positive trend will continue in 2017 with Horizon Blue Cross Blue Shield recently issuing positive policy coverage for more than 3.8 million members. The recently obtained unique ConfirmMDx CPT code, effective January 2018, is expected to further streamline the Company's reimbursement efforts and significantly reduce collection periods. Further to the period end, MDxHealth was awarded a US Government Services Administration (GSA) contract.  This contract is critical to making ConfirmMDx widely accessible to authorized government customers. The Company's continued investments in demonstrating clinical utility and improved patient care from use of ConfirmMDx has resulted in publication in 10 clinical scientific publications and presentations, including in the journal The Prostate. Scientific data were also presented at several urology conferences across the US, including the Annual Meeting of the American Urology Association (AUA) in San Diego, and the Symposium of the American Society of Clinical Oncology Genitourinary Cancers Symposium (ASCO GU) in San Francisco. Successful global commercial launch of first liquid biopsy test SelectMDx. Second liquid biopsy test AssureMDx for Bladder Cancer set for 2017 full commercial launch Following the European launch in late 2015, the Company's proprietary liquid biopsy test SelectMDx was commercially launched in the US in March 2016. This non-invasive urine-based test identifies men at risk for clinically significant prostate cancer who may benefit from an initial prostate biopsy or magnetic resonance imaging (MRI). The validation study published in European Urology confirms the superior performance compared to other commonly used biomarker tests and risk calculators. The enrollment of the prospective 4M clinical study on 600 subjects evaluating the synergy of MRI and SelectMDx is complete and results are expected to be published in the course of 2017. In the US, SelectMDx is marketed as a Laboratory Developed Test (LDT) through MDxHealth's sales force. Outside the US, the Company pursues a direct sales strategy in the Benelux, Germany and Italy, complemented by European and global distributors and commercial lab partners. The Company concluded several agreements for the distribution of SelectMDx, covering various countries in Europe, Central and Latin America, Israel and most recently Hong Kong and Macao China. Outside the US, the test is currently performed at the Company's ISO 13485:2016 certified lab in Nijmegen, The Netherlands. Volumes for SelectMDx since its US launch in 2016 were nearly 3.5 times higher compared to the first year for ConfirmMDx, representing 17% of the total test volume for 2016. The fast uptake of this test by urologists and their patients has already resulted in 11 payor contracts in the first year. In the last quarter of 2016, the Company beta launched its second liquid biopsy test, AssureMDx for Bladder Cancer. The launch followed the publication of two clinical validation studies underscoring the test's clinical validity. AssureMDx has a very high negative predictive value of 99% and may significantly reduce unnecessary invasive and costly cystoscopy procedures. With 700,000 cystoscopy procedures performed in the US every year, AssureMDx addresses a potential $300 million market in the US, and is scheduled for commercial launch in the US as an LDT by the end of the first half of 2017. MDxHealth has built a robust portfolio of proprietary biomarkers for molecular assays for areas such as colorectal, lung and brain cancers. In addition to its proprietary offering in uro-oncology, the Company leverages this non-core portfolio of biomarkers and technologies through various licensing agreements. In 2016, MDxHealth granted a non-exclusive worldwide license for its methylation-specific PSR (MSP) technology to QIAGEN for use in its diagnostic cervical cancer assay, QIAsure. Key unaudited consolidated figures for the financial year ended December 31, 2016 (thousands of US dollars, except number of shares and per share data): Total revenues for the year ended December 31, 2016 amounted to $30 million, an increase of 70% compared to total revenues of $17.6 million a year earlier and included $25 million of product and service income. The strong growth in the US contributed $24.4 million or 82% of total revenue. Non-US revenues included initial sales of SelectMDx, milestone payments and royalties from license deals, and came in at $5.5 million, up 133% or $2.4 million compared to 2015. Total net billings for 2016 amounted to $49.3 million. However, the Company only recognizes revenue when there is reasonable evidence that the test will effectively be reimbursed. As a result, a significant portion of the revenue is only recognized when the payment is collected, leaving a significant portion of invoiced amounts unrecognized in 2016. The deferral of part of the revenue gradually decreases as the Company concludes firm agreements for reimbursement with a growing number of payors. Cost of goods sold for 2016 came in at $10.1 million, compared to $6.9 million in 2015. Increased revenues resulted in a gross profit of $19.9 million, while a sustained focus on operational efficiencies yielded an improvement of the gross profit margin from 60.9% in 2015 to 66.3% in 2016. Operating expenses for the year ended December 31, 2016 amounted to $32.7 million, an increase of $7.6 million or 30.2%. The year-on-year increase is mainly attributable to the acquisition of NovioGendix (renamed to MDxHealth BV), which was only included for one quarter in 2015. Furthermore, MDxHealth invested in the build-out of the organisation to support the global commercial launch of SelectMDx. MDxHealth adopts a direct sales strategy for SelectMDx in Benelux, Germany and Italy, supported by European and global distributors and commercial lab partners. The Company appointed a global commercial team to cover business development and direct sales. Finally, the Company continued to validate the clinical utility of its expanded offering through clinical trials and publications. R&D spending remained level with last year despite the increased pace of newly developed product launches. EBITDA for the year improved by $2.5 million as the loss was reduced from $13.6 million in 2015 to $11.1 million. This improvement was partly offset by increased amortization charges, bringing the Company's net loss for 2016 to $13.2 million, $1.3 million better than in 2015. The increased amortization resulted from scheduled amortization of intangible assets associated with the acquisition of NovioGendix in 2015. Cash and cash equivalents stood at $30.8 million at the end of 2016 after having successfully raised $21.7 million (€20.4 million) in a private placement of 4,526,962 new shares at €4.50 ($4.99) per share. The number of outstanding shares at December 31 was 49,796,595. Increased private payor adoption and a sustained focus on reimbursement has helped to improve working capital throughout 2016. Cash used by operations amounted to $16.6 million, compared to $14.4 million in 2015, and included cash collections of $19.7 million, a 61% increase year-on-year. The award of a unique CPT code by the AMA, which will become effective January 1, 2018, is expected to significantly shorten collection periods from both Medicare and private payors. MDxHealth is committed to maintaining its focus on its strategic priorities during 2017. These include: MDxHealth will host a conference call today at 15:00 CET / 14:00 GMT / 09:00 EST / 06:00 PT to discuss its Financial Year 2016 results. To access the conference call, please dial one of the appropriate numbers below quoting the conference ID 69886101. The call will be conducted in English and a replay will be available for 30 days. The presentation will be made available on the investors section of the MDxHealth website shortly before the call and can be accessed at: http://mdxhealth.com/investors. The Company's statutory auditor, BDO Bedrijfsrevisoren Burg. Ven. CBVA, has confirmed that its audit procedures with respect to the Company's consolidated financial statements, prepared in accordance with the International Financial Reporting Standards as adopted in the European Union, have been substantially completed, that these procedures have not revealed any material adjustments that would have to be made to the accounting information derived from the Company's consolidated financial information that is included in this press release, and that it intends to issue an unqualified opinion. The condensed Consolidated Statement of Comprehensive Income may be found on the Company's website at www.mdxhealth.com. The full Annual Report is expected to be made available to the public via the Company's website in April 2017. MDxHealth is a multinational healthcare company that provides actionable molecular diagnostic information to personalize the diagnosis and treatment of cancer. The Company's tests are based on proprietary genetic, epigenetic (methylation) and other molecular technologies and assist physicians with the diagnosis of urologic cancers, prognosis of recurrence risk, and prediction of response to a specific therapy. MDxHealth's European headquarters are in Herstal, Belgium, with laboratory operations in Nijmegen, The Netherlands, and US headquarters and laboratory operations based in Irvine, California. For more information visit mdxhealth.com and follow us on Twitter at: twitter.com/mdxhealth. This press release contains forward-looking statements and estimates with respect to the anticipated future performance of MDxHealth and the market in which it operates. Such statements and estimates are based on assumptions and assessments of known and unknown risks, uncertainties and other factors, which were deemed reasonable but may not prove to be correct. Actual events are difficult to predict, may depend upon factors that are beyond the company's control, and may turn out to be materially different. MDxHealth expressly disclaims any obligation to update any such forward-looking statements in this release to reflect any change in its expectations with regard thereto or any change in events, conditions or circumstances on which any such statement is based unless required by law or regulation.  This press release does not constitute an offer or invitation for the sale or purchase of securities or assets of MDxHealth in any jurisdiction. No securities of MDxHealth may be offered or sold within the United States without registration under the US Securities Act of 1933, as amended, or in compliance with an exemption therefrom, and in accordance with any applicable US securities laws. NOTE: The MDxHealth logo, MDxHealth, ConfirmMDx, SelectMDx, AssureMDx and PredictMDx are trademarks or registered trademarks of MDxHealth SA. All other trademarks and service marks are the property of their respective owners.


News Article | February 15, 2017
Site: spaceref.com

Like cosmic lighthouses sweeping the universe with bursts of energy, pulsars have fascinated and baffled astronomers since they were first discovered 50 years ago. In two studies, international teams of astronomers suggest that recent images from NASA's Chandra X-ray Observatory of two pulsars -- Geminga and B0355+54 -- may help shine a light on the distinctive emission signatures of pulsars, as well as their often perplexing geometry. Pulsars are a type of neutron star that are born in supernova explosions when massive stars collapse. Discovered initially by lighthouse-like beams of radio emission, more recent research has found that energetic pulsars also produce beams of high energy gamma rays. Interestingly, the beams rarely match up, said Bettina Posselt, senior research associate in astronomy and astrophysics, Penn State. The shapes of observed radio and gamma-ray pulses are often quite different and some of the objects show only one type of pulse or the other. These differences have generated debate about the pulsar model. "It's not fully understood why there are variations between different pulsars," said Posselt. "One of the main ideas here is that pulse differences have a lot to do with geometry -- and it also depends on how the pulsar's spin and magnetic axes are oriented with respect to line of sight whether you see certain pulsars or not, as well as how you see them." Chandra's images are giving the astronomers a closer than ever look at the distinctive geometry of the charged particle winds radiating in X-ray and other wavelengths from the objects, according to Posselt. Pulsars rhythmically rotate as they rocket through space at speeds reaching hundreds of kilometers a second. Pulsar wind nebulae (PWN) are produced when the energetic particles streaming from pulsars shoot along the stars' magnetic fields, form tori -- donut-shaped rings -- around the pulsar's equatorial plane, and jet along the spin axis, often sweeping back into long tails as the pulsars' quickly cut through the interstellar medium. "This is one of the nicest results of our larger study of pulsar wind nebulae," said Roger W. Romani, professor of physics at Stanford University and principal investigator of the Chandra PWN project. "By making the 3-D structure of these winds visible, we have shown how one can trace back to the plasma injected by the pulsar at the center. Chandra's fantastic X-ray acuity was essential for this study, so we are happy that it was possible to get the deep exposures that made these faint structures visible." A spectacular PWN is seen around the Geminga pulsar. Geminga -- one of the closest pulsars at only 800 light-years away from Earth -- has three unusual tails, said Posselt. The streams of particles spewing out of the alleged poles of Geminga -- or lateral tails -- stretch out for more than half a light-year, longer than 1,000 times the distance between the Sun and Pluto. Another shorter tail also emanates from the pulsar. The astronomers said that a much different PWN picture is seen in the X-ray image of another pulsar called B0355+54, which is about 3,300 light-years away from Earth. The tail of this pulsar has a cap of emission, followed by a narrow double tail that extends almost five light-years away from the star. While Geminga shows pulses in the gamma ray spectrum, but is radio quiet, B0355+54 is one of the brightest radio pulsars, but fails to show gamma rays. "The tails seem to tell us why that is," said Posselt, adding that the pulsars' spin axis and magnetic axis orientations influence what emissions are seen on Earth. According to Posselt, Geminga may have magnetic poles quite close to the top and bottom of the object, and nearly aligned spin poles, much like Earth. One of the magnetic poles of B0355+54 could directly face the Earth. Because the radio emission occurs near the site of the magnetic poles, the radio waves may point along the direction of the jets, she said. Gamma-ray emission, on the other hand, is produced at higher altitudes in a larger region, allowing the respective pulses to sweep larger areas of the sky. "For Geminga, we view the bright gamma ray pulses and the edge of the pulsar wind nebula torus, but the radio beams near the jets point off to the sides and remain unseen," Posselt said. The strongly bent lateral tails offer the astronomers clues to the geometry of the pulsar, which could be compared to either jet contrails soaring into space, or to a bow shock similar to the shockwave created by a bullet as it is shot through the air. Oleg Kargaltsev, assistant professor of physics, George Washington University, who worked on the study on B0355+54, said that the orientation of B0355+54 plays a role in how astronomers see the pulsar, as well. The study is available online in arXiv. "For B0355+54, a jet points nearly at us so we detect the bright radio pulses while most of the gamma-ray emission is directed in the plane of the sky and misses the Earth," said Kargaltsev. "This implies that the pulsar's spin axis direction is close to our line-of-sight direction and that the pulsar is moving nearly perpendicularly to its spin axis." Noel Klingler, a graduate research assistant in physics, George Washington University, and lead author of the B0355+54 paper, added that the angles between the three vectors -- the spin axis, the line-of-sight, and the velocity -- are different for different pulsars, thus affecting the appearances of their nebulae. "In particular, it may be tricky to detect a PWN from a pulsar moving close to the line-of-sight and having a small angle between the spin axis and our line-of-sight," said Klingler. In the bow-shock interpretation of the Geminga X-ray data, Geminga's two long tails and their unusual spectrum may suggest that the particles are accelerated to nearly the speed of light through a process called Fermi acceleration. The Fermi acceleration takes place at the intersection of a pulsar wind and the interstellar material, according to the researchers, who report their findings on Geminga online and in the current issue of Astrophysical Journal. Although different interpretations remain on the table for Geminga's geometry, Posselt said that Chandra's images of the pulsar are helping astrophysicists use pulsars as particle physics laboratories. Studying the objects gives astrophysicists a chance to investigate particle physics in conditions that would be impossible to replicate in a particle accelerator on earth. "In both scenarios, Geminga provides exciting new constraints on the acceleration physics in pulsar wind nebulae and their interaction with the surrounding interstellar matter," she said. * "Deep Chandra Observations of the Pulsar Wind Nebula Created by PSR B0355+54," Noel Klingler et al., 2016 Dec. 20, Astrophysical Journal [http://iopscience.iop.org/article/10.3847/1538-4357/833/2/253, preprint: https://arxiv.org/abs/1610.06167]. * "Geminga's Puzzling Pulsar Wind Nebula," B. Posselt et al., 2017, to appear in Astrophysical Journal [http://apj.aas.org, preprint: https://arxiv.org/abs/1611.03496]. Other team members include George C. Pavlov, senior scientist in astronomy and astrophysics, Penn State; Pat O. Slane, lecturer and senior astrophysicist, Harvard Smithsonian Center for Astrophysics; Roger Romani, professor of physics, Stanford University; Niccolo Bucciantini, permanent researcher, INAF Osservatorio Astorfisico di Arcetri; Andrei M. Bykov, head of the Laboratory for High Energy Astrophysics, Ioffe Physical-Technical Institute; Martin C. Weisskopf, project scientist, NASA/Marshall Space Flight Center; Stephen Chi-Yung Ng, assistant professor of physics, University of Hong Kong. Additional team members for the study on B0355+54 include Blagoy Rangelov, postdoctoral researcher, George Washington University; Tea Temim, JWST Support Scientist, Space Telescope Science Institute; Douglas A. Swartz, research scientist, Marshall Space Flight Center and Rolf Buehler, staff scientist, DESY Zeuthen. NASA and the Russian Science Foundation supported this work. Please follow SpaceRef on Twitter and Like us on Facebook.


While studying age-associated dendritic restructuring in C. elegans neurons7, we noticed that fluorescent signals originating from neurons sometimes appeared situated outside of the cell in defined vesicle-like structures that we call exophers (Fig. 1a–c, Extended Data Figs 1a–c, 2g). We first characterized exophers associated with the six gentle touch receptor neurons, for which cell bodies and dendrites are easily visualized. We found that exophers are comparable in size (average diameter 3.8 μM) to neuronal somas (Extended Data Fig. 1d). The size of the vesicles, the morphological stages in their biogenesis (Fig. 1a–c), and the genetic requirements for their production (Extended Data Table 1a) distinguish them from much smaller exosomes (around 30–100 nm; Extended Data Table 2 compares exophers to characterized extracellular vesicles). Neuronal exophers do not seem to result from classical cell division: (1) exophers did not stain with the nuclear DNA indicator DAPI (Fig. 1b); (2) cell division-inhibiting hydroxyurea8 did not change exopher levels (n > 30 per trial, three trials); and (3) RNA interference (RNAi)-mediated disruption of cell cycle genes did not change exopher detection (Extended Data Table 1b). We found that exopher production is not restricted to a specific transgene reporter or line (examples in Fig. 1, Extended Data Fig. 1). Amphid neurons that are dye-filled via openings to the outside environment9 (Extended Data Fig. 1e, f) can produce exophers, confirming that exophers can form under native physiological cellular conditions. Exopher production differs markedly among the six touch receptor neurons, with ALMR neurons producing exophers most frequently (Fig. 1d). Many neuronal types can produce exophers, including dopaminergic PDE and CEP neurons (Extended Data Fig. 1g, h), FLP neurons (not shown) and sensory ASER neurons (Extended Data Fig. 1i). Time-lapse analyses (Supplementary Videos 1, 2) revealed that exophers typically arise from the soma by asymmetrically amassing labelled protein to create a balloon-like extrusion via a pinching off event; the exopher compartment then moves outward from the neuronal cell body (extrusion approximately 15–100 min; Fig. 1a, Extended Data Fig. 1a). The plasma membrane reporter P PH(plcDelta)::GFP (Extended Data Fig. 2a) and electron microscopy data (Extended Data Fig. 2) confirm that exophers are membrane-bound. Exophers can initially remain connected to the soma by a thin thread-like tube (Fig. 1c) that allows the transfer of tagged proteins and calcium into the attached exopher compartment (Extended Data Figs 1a, 3, Supplementary Video 2). Exophers ultimately disconnect from the originating neuronal soma (Extended Data Fig. 3). Time-lapse studies indicated that aggregating mCherry often appeared preferentially concentrated into exophers, and neurons expressing the huntingtin (Htt) protein with a neurotoxic polyglutamine tract of 128 repeats (Htt-Q128) could also concentrate and extrude this aggregating protein in exophers (Fig. 2a, b). We therefore further queried the relationship of aggregating or toxic protein expression to exopher production. Strains expressing Q128 (toxic, with high levels of apparent aggregation10, 11) produced significantly more exophers than strains that did not express polyQ or that expressed Htt-Q19 (non-toxic and low aggregation) (Fig. 2c). Likewise, aggregating mCherry lines exhibited higher average exopher numbers over adult life than lines expressing soluble green fluorescent protein (GFP) (see Fig. 2d). High aggregate load in individual neurons was predictive of increased exopher production on the following day (Fig. 2e). Conversely, mCherry RNAi reduced the number of exophers by approximately one-half in a line producing aggregating mCherry (Fig. 2f). Although our studies cannot determine the relative contribution of aggregate load from protein expression levels, they suggest that proteostatic challenges increase exopher production. Consistent with a potential role for exophers in the elimination of potentially harmful neuronal contents, the expression of amyloid-forming human Alzheimer’s disease fragment amyloid-β in ASER neurons increases exopher numbers (Fig. 2g). Our combined observations on exopher formation, contents and frequency of detection suggest that exophers preferentially include aggregated, excess, or otherwise neurotoxic proteins for removal. To address the hypothesis that aggregation-prone proteins might be selectively extruded in exophers, we constructed a line that expressed both an aggregation-prone mCherry (Is[P mCh1]) and a non-aggregating GFP (Is[P GFP]) and compared the red and green fluorescence distribution between exophers and somas (example in Fig. 2h, data in Fig. 2i). In 22 out of 23 exophers, we found higher relative levels of mCherry in the exopher, and higher relative levels of GFP in the soma. Neurons appear to extrude aggregation-prone mCherry preferentially compared with soluble GFP, suggesting that deleterious materials are identified and sorted for export during exopher-genesis. To investigate whether proteostatic challenges enhance the exopher production response, we manipulated the in vivo protein-folding milieu. We found a roughly sixfold increase in exopher production in an hsf-1(sy441) mutant deficient in the core proteostasis transcription factor HSF-1 (and therefore deficient in chaperone expression) (Fig. 3a). We impaired autophagy by treating animals with the pharmacological inhibitor spautin-1 and by RNAi knockdown (lgg-1, atg-7, bec-1, lgg-1/2) in a strain expressing aggregation-prone mCherry, and measured a significant increase in exopher incidence (Fig. 3b, c). Impairment of proteasome activity with the inhibitor MG132 on strain Is[P mCh1] also increased exopher production (Fig. 3d). Given that inhibiting several facets of proteostasis increases exopher extrusion, we suggest that exophers may constitute a previously undescribed component of the proteostasis network, which may function as a backup or alternative response to rid cells of neurotoxic aggregates/proteins when proteostasis becomes overwhelmed by mounting intracellular proteotoxicity. Exopher production occurs with a notable bimodal distribution throughout adult life: exophers are most commonly observed at adult days A2–A3, diminish in abundance at A4–A8, and then reappear again later in life at approximately A10–A11 (Fig. 2d; similar young adult pattern with dye-filled amphid neurons, Extended Data Fig. 1f; and with a 1-day earlier onset in an hsf-1 mutant, Extended Data Fig. 1j). The distinctive temporal production profile suggests that conditions permissive for exopher production exist in young adulthood but can then be limited or remain below a threshold until late adulthood. The coincidence of the early peak with a transition in C. elegans young adult proteostasis management12, 13, 14 suggests that the first wave of exopher-genesis may serve as a normal component of an orchestrated proteostasis reset in young adulthood that involves the removal of neuronal debris generated during development; the later adult increase in exopher production may be the consequence of age-associated decline in proteostatic robustness. Rather than inducing neuronal death or dysfunction, exopher-genesis seems to be beneficial. First, in hundreds of longitudinal observations, we did not observe neuronal loss after exopher production: exophers are distinct from apoptotic bodies in their biogenesis (Fig. 1a, Extended Data Fig. 1a), and the soma of an exopher-producing neuron retains normal ultrastructural features (Extended Data Fig. 2e). Second, the relative functionality of proteotoxically stressed neurons that have generated exophers is increased compared with neurons that did not extrude exophers. In blinded studies of a line expressing cyan fluorescent protein (CFP)-tagged Q128, which progressively impairs touch sensation10, we found that midlife touch sensitivity is greater when ALMR had definitely produced an exopher at A2, as compared to age-matched siblings in which ALMR had not produced an exopher (Fig. 3e). Third, we identified pod-1 and emb-8 as polarity genes required in adults for exopher-genesis (Fig. 3f), and found that adult RNAi knockdown impaired midlife touch sensitivity (Fig. 3g). Although we cannot rule out that pod-1 and emb-8 RNAi interventions might generally disrupt adult neuronal function, taken together our data are consistent with a model in which adult neurons that do not make exophers become functionally compromised compared to those neurons that extruded offending contents. Overall, adult neurons seem to be healthier after a considerable expulsion of potentially toxic contents. Considering the large apparent volume of exophers, we proposed that they might include organelles. Indeed, both lysosomes (Extended Data Fig. 4) and mitochondria (Fig. 4a, b, Extended Data Fig. 5) can be extruded in exophers. Mitochondrially localized GFP reporters revealed mitochondrial inclusion in budding and dissociated exophers, with punctate or filamentous morphology typical of adult mitochondrial networks (Fig. 4a, Extended Data Fig. 5a–c). To address whether impairing mitochondrial quality enhances the production of exophers, we genetically manipulated the mitophagy mediator dct-1 (homologue of mammalian BNIP3), the human Parkinson’s disease homologues pink-1 (PINK)15 and pdr-1 (PARK2)16 implicated in mitochondrial maintenance, and the mitochondrial unfolded protein response gene ubl-5 (ref. 17) (Fig. 4c, d). We conclude that several genetic approaches that impair mitochondria can increase exopher-genesis. To address the hypothesis that stressed or damaged mitochondria might be preferentially segregated to exophers, we used the mitochondrial reporter mitoROGFP, which changes its peak excitation wavelength from around 405 nm (oxidized) to 476 nm (reduced) according to the local oxidative environment18, 19. We find a significant increase in the 405 nm (oxidized)/476 nm (reduced) excitation ratio of mitochondria in exophers compared to those in somas (Fig. 4e), roughly equivalent to the redox excitation ratio observed in C. elegans neurons subjected to H O -induced oxidative stress19. We confirmed higher oxidation scores using MitoTimer, an alternative reporter of mitochondrial matrix oxidation20 (Extended Data Fig. 5d). In addition, touch neurons of juglone-treated21 bzIs166[P mCherry]; zhsEx17[P mitoLS::ROGFP] animals had significantly higher numbers of mitochondria-including exophers than matched controls (Extended Data Fig. 5e). Although compromised mitochondrial health may impair neuronal proteostasis, thus increasing exopher production, our data establish that touch neurons can eject mitochondria via exophers, which raises the intriguing possibility that exopher-genesis may constitute a previously unappreciated removal-based mechanism of mitochondrial homeostasis. We next sought to determine the fate of the extruded exopher and its contents. With time, exopher fluorescence intensity diminishes or disappears (persistence times 1–12 h), possibly as exopher contents are degraded internally or digested by the neighbouring hypodermis that fully surrounds the touch neuron and has degradative capabilities. Disruption of the C. elegans apoptotic engulfment genes ced-1 (homologue of mammalian CD91, LRP1 and MEGF10, and fly Draper), ced-6 (GULP) and ced-7 (ABC1) increases the detection of ALMR neurons that have extruded several exophers (Fig. 5a, Extended Data Fig. 6a); however, the genetic manipulation of a parallel engulfment pathway comprising ced-2 (Crk-II), ced-5 (DOCK180), ced-10 (RAC1), ced-12 (ELMO) and psr-1 (PSR) did not change the frequency of exopher generation or the detection of multiple exophers. Moreover, we did not detect the apoptotic ‘eat-me’ signal phosphatidylserine on the exopher surface using a widely expressed phosphatidylserine-binding annexinV::GFP (0 out of 43 exophers; Extended Data Fig. 6b). Our data suggest that hypodermal recognition/degradation of exophers and their contents occurs by mechanisms that are at least in part distinct from the classical removal of apoptotic corpses, but involve the CED-1, CED-6 and CED-7 proteins. Electron microscopy studies also show that the hypodermis may mediate the degradation of at least some exopher contents (Extended Data Fig. 2d–f, h). The lack of a detectable phosphatidylserine signal on exophers raised the question as to whether at least some exopher contents might be destined to elude hypodermal degradation. Indeed, fluorescent mCherry protein that was originally expressed specifically in touch neurons, or fluorescent DiI loaded into dye-filling neurons, appeared later in distant scavenger coelomocytes (Fig. 5b–d, Extended Data Fig. 6c). Blocking coelomocyte uptake capacity by cup-4 mutation22 caused fluorescent particles to accumulate outside neurons, possibly within the pseudocoelom (body cavity; Extended Data Fig. 6d, e). We conclude that some exopher contents transit the hypodermal tissue to be released into the pseudocoelomic fluid, from which materials can later be taken up by distant coelomocytes. Exophers can therefore mediate transfer of neuronal materials to remote cells. Considerable excitement in the neurodegenerative disease field has been generated by the findings that mammalian neurons can extrude conformational disease proteins, including in Alzheimer’s, Parkinson’s and prion disease23. The production of exophers in C. elegans constitutes a newly identified mechanism by which neurons can transfer cellular material (preferentially neurotoxic species) to other cells. Notably, in a C. elegans muscle model of prion toxicity, offending prion proteins were transferred among muscle cells and ultimately localized to coelomocytes24. We speculate that the basic mechanism we document here may correspond to a conserved pathway for the transfer of toxic contents out of many cell types. In this regard, it may be noteworthy that mammalian aggregated poly-Q-expanded huntingtin can transfer between neurons via tunnelling nanotubes25, 26, 27 that resemble thin connections between C. elegans somas and exophers, and that neuronal polyQ in Drosophila is transferred to glia via a process that requires the CED-1 homologue, Draper28. Recent reports show that mitochondria can transfer out of specific cells to contribute positive roles (mesenchymal stem cells via tunnelling nanotubes29; astrocytes to neurons in a stroke model30), but our study underscores a generally underappreciated option for mitochondrial quality control: mitochondrial expulsion. The mitochondrial expulsion we report in C. elegans touch neurons has a notable mammalian counterpart: mouse mitochondria originating in retinal ganglion cells can be extruded into neighbouring astrocytes for degradation6 (with some similar morphology to C. elegans exophers; see fig. 1e of ref. 6). Although further study will be required to establish definitively the health status and fates of transferred mitochondria in the C. elegans model, it is tempting to speculate that transcellular degradation of mitochondria may be a more broadly used mechanism of mitochondrial quality control than currently appreciated, with associated potential importance in neuronal health. Overall, although further experiments are needed to determine the detailed mechanisms at play and validate the proposed functions of exophers in proteostasis and the removal of damaged organelles, we suggest that exopher production is a previously unrecognized mechanism for clearing out accumulating protein aggregates and dysfunctional organelles that threaten neuronal homeostasis (Extended Data Fig. 7). The analogous process in mammals could enable the transfer of misfolded protein and/or dysfunctional mitochondria to neighbouring cells, promoting human pathology in neurodegenerative disease if compromised. Mechanistic dissection of this new aspect of proteostasis and mitochondrial homeostasis should thus inform on fundamental mechanisms of neuronal maintenance and suggest targets for intervention in neurodegenerative disease.

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