la Guatemalan Black Howler Monkey By feedproxy.google.com Published On :: Fri, 03 Jul 2015 17:05:00 +0200 The endangered Guatemalan Black Howler (Alouatta pigra) (sometimes called the Yucatan Howler or Yucatan Black Howler) is one of many species of howler monkey, which is what is known as a ‘New World’ monkey. Its range is throughout the Yucatan Peninsula, and includes the areas of Mexico, Belize, and of course Guatemala. The Guatemalan Black Howler prefers to live in very lush areas, mostly sticking to all types of rain forests such as the semi-deciduous, lowland and evergreen. Of its cousins and relatives, the Guatemalan Black Howler is the largest, and is also one of the largest ‘New World’ monkeys (which include marmosets, owl monkeys, sakis, spider, and woolly monkeys). It weighs in at 25 lbs on average in males (11-12 kg) and 14 lbs for the females (6-7 kg). Their fur is usually black and their tails are very long, and prehensile (meaning it can grab and be used to hang from branches with). They also have specialized teeth for their diet of mostly leaves, along with the males possessing a larger hyoid bone located near the vocal chords, which enables their loud calls. The Guatemalan Black is a diurnal howler, which means it is active during the day and it sleeps at night, as well as being arboreal, meaning it dwells in the trees most of its life. They are a social species that lives in groups up to ten members strong, which helps in alerting, foraging, and general upkeep through grooming. Some groups can be as large as sixteen, while larger groups are plausible, however at these sizes it is unlikely to function as well as a smaller group. Their diets consist of mostly leaves, and fruits, however they will snack on a flower here and there and their favorite tree of all is the breadnut, which provides most food during some seasons. Not a particularly active species, the Guatemalan Black Howler would rather lounge about during the day; eating takes up a quarter of the day while moving locations for eating consists of only about a tenth of their daily activity. The rest of the day is devoted to socializing and grooming, with some other random antics. Females are old enough to have offspring by four years of age, while males may take up to eight years to reach maturity, and their total life-spans are an average of twenty years. The Guatemalan Black Howler’s binomial name (its species and genus) is Alouatta pigra, the Alouatta’s as a genus make up most of the Howler Monkeys, which are the largest of the New World Monkeys with but a few possible exceptions. Alouatta is home to all of the howler monkeys (ten species and ten subspecies), and belongs to the subfamily Alouattinae. Alouattinae belongs to the family Atelidae which is one of the four families of New World Monkeys; this includes the howler monkeys, along with spider monkeys, woolly monkeys, wooly spider monkeys, and Yellow-tailed Woolly Monkeys. Atelidae belongs to the Parvorder Platyrrhini, which contains all New World Monkeys, and includes Marmosets and Muriquis. Platyrrhini belongs to the infraorder Simiiformes, or ‘higher primates’, and this includes all New World and Old World monkeys from South America and Africa, and includes gibbons, great apes, and the family Hominidae of which we are members. Simiiformes belongs to the Suborder Haplorrhini, otherwise called dry-nosed primates; this includes all of the higher primates as well as Tarsiers. Haplorrhini belongs to the Order Primates, which is all related apes, monkeys, lorids, galagos, lemurs and human ancestors. Primates are in the class of Mammalia of the phylum Chordata in the Kingdom of Animalia. Fact The Guatemalan Black Howler is sympatric with another species, the Mantled Howler. Sympatric means that they share the same niche and territory, and encounter each other in the wild, they are also closely related. Warning The Guatemalan Black Howler is an Endangered Species, and is close to being classified as Critically Endangered if nothing is done to curb the loss of the species. In the next 30 years the IUCN expects to see a population loss of over 60%, making this species on the more endangered alive today. Major threats are habitat loss, poaching, and capture for use as ‘pets’. Full Article default
la Airhook Looks to Reinvent the Traditional Airplane Tray Table By feedproxy.google.com Published On :: Thu, 06 Jun 2019 14:17:52 +0000 A clever, versatile, well-made alternative to traditional airplane tray tables. The post Airhook Looks to Reinvent the Traditional Airplane Tray Table appeared first on Vagabondish. Full Article Gear Modern Vagabond Review airplane Tray Table
la Brighten Up Your After-Dark Road-Tripping with Lanmodo’s Vast Automotive Night Vision System By feedproxy.google.com Published On :: Mon, 02 Sep 2019 01:40:35 +0000 With a bright, 8.2-inch HD display and the flexibility to work in almost any vehicle, the Lanmodo Automotive Vast Night Vision System is a near-perfect alternative to factory-installed systems. The post Brighten Up Your After-Dark Road-Tripping with Lanmodo’s Vast Automotive Night Vision System appeared first on Vagabondish. Full Article Modern Vagabond
la BioLite HeadLamp 200 Is the Featherweight Head Lamp You’ll (Probably) Forget You’re Wearing By feedproxy.google.com Published On :: Fri, 06 Mar 2020 21:43:29 +0000 It's not the brightest headlamp on the market, but it's more than adequate for cooking, reading, or finding a pee tree in the dark. Plus, it's comfortable, easy-to-use, and USB rechargeable. The post BioLite HeadLamp 200 Is the Featherweight Head Lamp You’ll (Probably) Forget You’re Wearing appeared first on Vagabondish. Full Article Modern Vagabond Review Camping
la Employee-surveillance software is not welcome to integrate with Basecamp By feedproxy.google.com Published On :: Thu, 07 May 2020 20:17:17 +0000 We’ve been teaching people how to do remote work well for the better part of two decades. We wrote a whole book about the topic in 2013, called REMOTE: Office Not Required. Basecamp has been a remote company since day one, and our software is sold as an all-in-one toolkit for remote work. Yeah, we’re… keep reading Full Article Uncategorized
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la At a glance: Best picture films By www.bbc.co.uk Published On :: 2008-02-22T12:30:00 Part of the Going for gold promo for the BBC UK Homepage Full Article
la News: Glaciers surge to ocean By www.bbc.co.uk Published On :: 2008-02-26T13:30:00 Part of the Antarctic diary promo for the BBC UK Homepage Full Article
la My Faith in Nerds: Stronger Than Any Gelatinous Cube By feedproxy.google.com Published On :: Fri, 10 Sep 2010 20:01:57 +0000 dConstruct 2010: Merlin Mann - "Kerning, Orgasms & Those Goddamned Japanese Toothpicks" (NSFW) ME - Kerning, Orgasms And Those Goddamned Japanese Toothpicks on Huffduffer Download the audio | Huffduff it Let's be honest. I don't go...mmmm...places very often. I sit in this chair. I go to the Safeway with my daughter. Sometimes, I take the train downtown to get a haircut. I check the mail. But, by and large, like most nerds, I'm without question, a bit of a shut-in. Which makes it more than a little ironic that my first trip off the North American continent brought me all the way to Brighton, England's wonderful dConstruct Conference. Which wonderful conference placed me inside a very royal complex, alone on a very large stage, 90 seconds after being informed I'd better be entertaining, because I'd be conducting my oration on the same spot where, a scant 36 years earlier, ABBA had become international stars by singing an up-tempo number about giving up. So, y'know. No pressure. Commanded to this location by two of my web heroes, I was told I could speak about whatever I wanted. So, wow, to quote the ladies of ABBA, how could I ever refuse? Thus, I stood on that stage for over 35 minutes, rambling to 800 talented, creative people about Dungeons & Dragons, japanese toothpicks, torrenting Photoshop, as well as what I used to find myself doing after a long evening of shooting mutants in Stargate.1 But, mostly? Yes. Mostly, I stood on a stage thousands of miles from the chair from which I barely move, and I told a lot of really smart people that they were nerds. I also told them they should get out more. I swear: it made sense at the time. Some Serious Talent My talk about the challenges and opportunities of being a giant nerd seemed well received. Honestly, I'm very happy with how it turned out. But--oh, brother--was I ever up against some heavy hitters. Serious Lou Gehrig shit. I'll leave it to other, more eloquent folks to tell you what a wonderful day this was. But I will very much suggest you learn this for yourself by listening to the audio of the fantastic talks. Because every one of them is a corker. Additionally, like I said, Tom Coates put on one of the loveliest slide decks it's ever been my pleasure to see (56MB PDF). Great speakers, great hosts, wonderful attendees (who aren't above buying a yank a pint [thanks, everybody]). And, Thanks, dConstruct I have to admit, I'm kind of over conferences as a thing, which makes it even more crazy when I go to one, and it blows me out of the water with the care and quality of the event, the speakers, and the attendees. dConstruct was absolutely one of those blown-out-of-the-water events. (photo: happy.apple) As I learned over and over again--yes, like me--these folks are nerds. But, brother are they ever talented nerds who care and care. Which I just love so much. I'll take a nerdy bunch of fontdorks and cellists over a splashy mega-conference full of VC pitches and skanks pushing free Red Bull anytime. Anytime. dConstruct was simply a top-notch operation from end-to-end, and I'm insanely grateful that I was invited to participate. Thanks, Clearleft. And, you, the reader? If you get the chance next time, go. Heck, I might even leave this chair and go there, myself. Maybe. I suppose when Dr. Who's over, I could just let these 20-sided dice decide for me. Lemme see...what's my Armor Class and Hit Points...? Listen for Yourself2 dConstruct Podcast MARTY NEUMEIER - The Designful Company The Designful Company on Huffduffer Download the audio | Huffduff it BRENDAN DAWES - Boil, Simmer, Reduce Boil, Simmer, Reduce on Huffduffer Download the audio | Huffduff it DAVID MCCANDLESS - Information Is Beautiful Information Is Beautiful on Huffduffer Download the audio | Huffduff it SAMANTHA WARREN - The Power and Beauty of Typography The Power and Beauty of Typography on Huffduffer Download the audio | Huffduff it JOHN GRUBER - The Auteur Theory Of Design The Auteur Theory Of Design on Huffduffer Download the audio | Huffduff it HANNAH DONOVAN - Jam Session: What Improvisation Can Teach Us About Design Jam Session: What Improvisation Can Teach Us About Design on Huffduffer Download the audio | Huffduff it JAMES BRIDLE - The Value Of Ruins The Value Of Ruins on Huffduffer Download the audio | Huffduff it TOM COATES - Everything The Network Touches Everything The Network Touches on Huffduffer Download the audio | Huffduff it Kerning, Orgasms And Those Goddamned Japanese Toothpicks ME - Kerning, Orgasms And Those Goddamned Japanese Toothpicks on Huffduffer Download the audio | Huffduff it Hint: Number Three. ↩ Code for these was stolen wholesale from the dConstruct site. Jeremy, et al - don't hesitate to tell me if that's a problem.Srsly. ↩ ”My Faith in Nerds: Stronger Than Any Gelatinous Cube” was written by Merlin Mann for 43Folders.com and was originally posted on September 10, 2010. Except as noted, it's ©2010 Merlin Mann and licensed for reuse under CC BY-NC-ND 3.0. "Why a footer?" Full Article dConstruct Geeks Knowledge Workers Nerds Speaking NSFW
la Resolved: Stop Blaming the Pancake By feedproxy.google.com Published On :: Fri, 07 Jan 2011 16:22:44 +0000 In a classic bit from an early Seinfeld, Jerry and Elaine are at the airport, trying to pick up the rental car that Jerry had reserved. As usual, things go poorly and get awkward fast: Seinfeld - "Reservations" JERRY: I don't understand...I made a reservation. Do you have my reservation? AGENT: Yes, we do. Unfortunately, we ran out of cars. JERRY: But, the reservation keeps the car here. That's why you have the reservation. AGENT: I know why we have reservations. JERRY: I don't think you do. If you did, I'd have a car. See, you know how to take the reservation--you just don't know how to hold the reservation. And, that's really the most important part of the reservation...the holding. Anybody can just TAKE them. [grabs chaotically at air] And, how weirdly similar is that to our conflicted relationship with New Year's resolutions? In Seinfeldspeak? See, you know how to make the resolution, you just don't know how to keep the resolution. And, that's really the most important part of the resolution...the keeping. Anybody can just MAKE them! Oversimplified? Probably. But, ask yourself. Why this? And, why now? Or, why again? Welcome to Resolvers Anonymous: I'm 'Merlin M.' A few years ago, I shared a handful of stories on the failures that have led to my own cynicism about the usefulness of life-inverting resolutions. Because, yeah, I've historically been a big resolver. Here's what I said when I first suggested favoring "Fresh Starts and Modest Changes" over reinventions: Download MP3 of "Fresh Starts & Modest Changes" Five years on, I think I probably feel even more strongly about this. Partly because I've watched and read and heard the cyclical lamentations of folks who decided to use superficial totems (like new calendars) as an ad hoc coach and prime mover. And, partly because, in my capacity as a makebelieve productivity expert, I continue to see how self-defeating it is to pretend that past can ever be less than prologue--that we can each ignore yesterday's weather if we really wish hard enough for a sun-drenched day at the beach. It simply doesn't work. Companies that think they'll be Google for buying bagels. Writers who think they'll get published if they order a new pen. Obese people who think they'll become marathon runners if they pick up some new running shoes. And, regular old people with good hearts who continue to confuse new lives with new clothes. Has this worked before? Can you look back on a proud legacy of successful New Year's resolutions that would suggest you're making serious progress by repeatedly making a list about fundamental life changes while slamming prosecco and wearing a pointy paper hat? My bet is that most people who are seeing the kind of change and growth and improvement that sticks tend to avoid these sorts of dramatic, geometric attempts to leap blindly toward the mountain of perfection. I'll go further and say that the repeated compulsion to resolve and resolve and resolve is actually a terrific marker that you're not really ready to change anything in a grownup and sustainable way. You probably just want another magic wand. Otherwise you'd already be doing the things you've resolved to do. You'd already be living those changes. And, you'd already be seeing actual improvements rather than repeatedly making lists of all the ways you hope your annual hajj to the self-improvement genie will fix you. Then, of course, we make things way worse by blaming everything on our pancakes. Regarding "The First Pancake Problem" Anyone who's ever made America's favorite round and flat breakfast food is familiar with the phenomenon of The First Pancake. No matter how good a cook you are, and no matter how hard you try, the first pancake of the batch always sucks. It comes out burnt or undercooked or weirdly shaped or just oddly inedible and aesthetically displeasing. Just ask your kids. At least compared to your normal pancake--and definitely compared to the far superior second and subsequent pancakes that make the cut and get promoted to the pile destined for the breakfast table--the first one's always a disaster. I'll leave it to the physicists and foodies in the gallery to develop a unified field theory on exactly why our pancake problem crops up with such unerring dependability. But I will share an orthogonal theory: you will be a way happier and more successful cook if you just accept that your first pancake is and always will be a universally flukey mess. But, that shouldn't mean you never make another pancake. So Loud. Then, So Quiet. I offer all of this because today is January 7th, gang. And, for the past week, all over the web, legions of well-intentioned and seemingly strong-willed humans have been declaring their resolved intention to make this a year of more and better metaphorical pancakes. And, like clockwork--usually around today or maybe tomorrow--a huge cohort of those cooks will begin to abandon their resolve and go back to thinking all their pancakes have to suck. Just because that first one failed. And, as is the case every year, online and off, there won't be nearly as many breathless updates to properly bookend how poorly our annual ritual of aspirational change has fared. Which is instructive. Not because new year's resolutions are a universally bad idea. And, not because Change is Bad. And, not because we should be embarrassed about occasionally falling short of our own (frequently unreasonable) aspirations. I suspect we tout the resolution, but whisper the failure because we blame the cook. Or, worse, fingers point toward the pancake. Instead of just admitting that the resolution itself was simply unrealistic or fundamentally foreign. And, that's a shame. Remember, there's no "I" in "unreasonable" Granted, I'm merely re-repeating a point I've struggled to make (to both others and myself) for years now. But, it will bear repeating every January in perpetuity. Resist the urge to pin the fate of things you really care about to anything that's not truly yourself. The "yourself" who has a real life with complicated demands. The "yourself" who's going to face a hard slog trying to fold a new life out of a fresh calendar. Calendars are just paper and staples. They can't make you care. And they can't help you spin around like Diana Prince, and instantly turn into Wonder Woman. Especially, if you're not already a hot and magical Amazon princess. First, be reasonable. Don't set yourself up for failure by demanding things that you've never come close to achieving before. I realize this is antithetical to most self-improvement bullshit, but that's exactly the point. If you were already a viking, you wouldn't need to build a big boat. Start with where you are right now. Not with where you wish you'd been. Also, accept that the first pancake will always suck. Hell, if you've never picked up a spatula before, be cool with the fact that your first hundred pancakes might suck. This is, as I've said, huge. Failure is the sound of beginning to suck a little less. And, finally, also be clear about the sanity of the motivations underlying your expectations--step back to observe what's truly broken, derive a picture of incremental success that seems do-able, and really resolve to do whatever you can realistically do to actually get better. Rather than "something something I suddenly become all different." At this point, you have logistical options for both execution and troubleshooting: Make a modest plan that you can envision actually doing without upending your real life; Build more sturdy scaffolding for sticking with whatever plan you've chosen; Make a practice of learning to not mind the duds--including those messed-up first pancakes; Or--seriously?--just accept that you never really cared that much about making breakfast in the first place. Care is not optional. Otherwise, really, you'd never need to resolve to do anything. You'd already just be cooking a lot. Instead of being all mad and depressed about not cooking. But, please. All I really ask of you. Don't blame the pancake. It's not really the pancake's fault. Like me, the pancake just wants you to be happy. This and every other new year. ”Resolved: Stop Blaming the Pancake” was written by Merlin Mann for 43Folders.com and was originally posted on January 07, 2011. Except as noted, it's ©2010 Merlin Mann and licensed for reuse under CC BY-NC-ND 3.0. "Why a footer?" Full Article change is hard resolutions self-improvement
la REPORT: US slams new restrictions on journalists from this country By dennismichaellynch.com Published On :: Sat, 09 May 2020 17:23:28 +0000 The DML News App offers the best in news reporting. The post REPORT: US slams new restrictions on journalists from this country appeared first on Dennis Michael Lynch. Full Article News Feed Powered by DMLNewsApp.com
la Alliston Honda plant to resume production with new safety protocols on production lines By barrie.ctvnews.ca Published On :: Fri, 8 May 2020 12:06:03 -0400 After seven weeks, Honda Canada in Alliston will gradually begin operations next week. Full Article
la Mother Nature blankets parts of Muskoka with spring snowfall By barrie.ctvnews.ca Published On :: Fri, 8 May 2020 15:03:52 -0400 The calendar may say May, but that didn't stop Mother Nature from blasting some wintry weather in Muskoka on Friday. Full Article
la Legal landscape murky for B.C. workers and employers during pandemic By bc.ctvnews.ca Published On :: Fri, 8 May 2020 19:39:00 -0700 Labour laws haven’t changed in our province, but legal experts are already urging B.C. employers to be flexible and reasonable — while warning employees they may not be legally protected if they refuse work during the pandemic. Full Article
la BREAKING: Sen Marsha Blackburn Introduces Stop COVID Act…Allowing US Citizens To Sue Communist China For Damage They’ve Inflicted On Our Nation By 100percentfedup.com Published On :: Wed, 06 May 2020 16:56:18 +0000 The following article, BREAKING: Sen Marsha Blackburn Introduces Stop COVID Act…Allowing US Citizens To Sue Communist China For Damage They’ve Inflicted On Our Nation, was first published on 100PercentFedUp.com. Yesterday, Senator Marsha Blackburn (R-TN), along with Senator Martha McSally (R-AZ) introduced the Stop COVID Act, giving Americans the ability to sue Communist China for the damage they’ve inflicted on our nation. Senator Blackburn appeared on Fox News with host Judge Jeanine where she explained the act to Jeanine Pirro. Blackburn told the Fox News […] Continue reading: BREAKING: Sen Marsha Blackburn Introduces Stop COVID Act…Allowing US Citizens To Sue Communist China For Damage They’ve Inflicted On Our Nation ... Full Article Featured Politics
la BREAKING: 43-Yr-Old Father Of 9, Dollar Store Sec Guard, Shot DEAD While Allegedly Trying To Enforce MI Gov Whitmer’s Face Covering Exec Order By 100percentfedup.com Published On :: Wed, 06 May 2020 18:27:38 +0000 The following article, BREAKING: 43-Yr-Old Father Of 9, Dollar Store Sec Guard, Shot DEAD While Allegedly Trying To Enforce MI Gov Whitmer’s Face Covering Exec Order, was first published on 100PercentFedUp.com. Will the mainstream media report about the senseless death of a 43-year-old father of nine, who would have been alive today if it were not for VP wannabe, Gretchen Whitmer's executive order that forced him to respond to a customer not wearing a face mask? Continue reading: BREAKING: 43-Yr-Old Father Of 9, Dollar Store Sec Guard, Shot DEAD While Allegedly Trying To Enforce MI Gov Whitmer’s Face Covering Exec Order ... Full Article Featured Left News Politics
la BREAKING: MI Lawmakers File Lawsuit Challenging Governor’s “Improper” and “Invalid” Emergency Orders: “We’ve attempted to partner with our governor, but she’s rejected” By 100percentfedup.com Published On :: Wed, 06 May 2020 20:54:20 +0000 The following article, BREAKING: MI Lawmakers File Lawsuit Challenging Governor’s “Improper” and “Invalid” Emergency Orders: “We’ve attempted to partner with our governor, but she’s rejected”, was first published on 100PercentFedUp.com. In addition to crushing Michigan's economy, the governor's reckless, one-size-fits-all executive orders are harming an untold number of Michigan citizens. Continue reading: BREAKING: MI Lawmakers File Lawsuit Challenging Governor’s “Improper” and “Invalid” Emergency Orders: “We’ve attempted to partner with our governor, but she’s rejected” ... Full Article Featured Politics
la Attorney for Jailed Dallas Salon Owner Shelley Luther Slams Obama Loyalist Judge as a “Tiny Tyrant” By 100percentfedup.com Published On :: Thu, 07 May 2020 02:33:27 +0000 The following article, Attorney for Jailed Dallas Salon Owner Shelley Luther Slams Obama Loyalist Judge as a “Tiny Tyrant”, was first published on 100PercentFedUp.com. Warren Norred, attorney for Shelley Luther, joined Tucker Carlson to discuss the decision by an Obama loyalist Texas judge to jail his client for reopening her salon during the COVID-19 pandemic. Norred said the judge has exposed himself for the “tiny tyrant” that he is. The judge jailed Luther and even fined her $7,000. Texas […] Continue reading: Attorney for Jailed Dallas Salon Owner Shelley Luther Slams Obama Loyalist Judge as a “Tiny Tyrant” ... Full Article Featured Government Politics
la Texas Gov Abbott Frees Salon Owner Shelley Luther: “Criminals shouldn’t be released to prevent COVID-19 just to put business owners in their place” By 100percentfedup.com Published On :: Thu, 07 May 2020 15:50:22 +0000 The following article, Texas Gov Abbott Frees Salon Owner Shelley Luther: “Criminals shouldn’t be released to prevent COVID-19 just to put business owners in their place”, was first published on 100PercentFedUp.com. Texas Governor Greg Abbott just changed the coronavirus order to free Salon A La Mode owner Shelley Luther from jail. Abbott tweeted out a comment about the poor treatment of the business owner: Throwing Texans in jail whose biz’s shut down through no fault of their own is wrong. I am eliminating jail for violating […] Continue reading: Texas Gov Abbott Frees Salon Owner Shelley Luther: “Criminals shouldn’t be released to prevent COVID-19 just to put business owners in their place” ... Full Article Breaking Featured Government Politics
la BREAKING: Michigan’s Radical Governor Extends Lockdown AGAIN…Only One Day After Church Leaders and MI Lawmakers File Suit Against Her By 100percentfedup.com Published On :: Thu, 07 May 2020 22:53:35 +0000 The following article, BREAKING: Michigan’s Radical Governor Extends Lockdown AGAIN…Only One Day After Church Leaders and MI Lawmakers File Suit Against Her, was first published on 100PercentFedUp.com. Michigan was the first state to see protests against draconian lockdown measures by an overreaching governor. On April 15, the Michigan Conservative Coalition organized an in-vehicle protest in Lansing, MI that drew an estimated 15,000 vehicles. Since that time, several protests by Michigan workers who are being devastated by the one-size-fits-all shut down of the […] Continue reading: BREAKING: Michigan’s Radical Governor Extends Lockdown AGAIN…Only One Day After Church Leaders and MI Lawmakers File Suit Against Her ... Full Article Featured Politics
la Mother of the Year, Michelle Obama Explains How Having Kids Was A “Concession”… Had To Give Up Her “Aspirations and Dreams” [VIDEO] By 100percentfedup.com Published On :: Fri, 08 May 2020 00:05:56 +0000 The following article, Mother of the Year, Michelle Obama Explains How Having Kids Was A “Concession”… Had To Give Up Her “Aspirations and Dreams” [VIDEO], was first published on 100PercentFedUp.com. While campaigning for her community organizer turned presidential candidate husband, Barack, Michelle Obama told a crowd of his supporters in Milwaukee, Wisconsin that for the first time in her life, she was proud to be an American. Four years later, Michelle Obama was a keynote speaker at the DNC convention, where she told Democrats how […] Continue reading: Mother of the Year, Michelle Obama Explains How Having Kids Was A “Concession”… Had To Give Up Her “Aspirations and Dreams” [VIDEO] ... Full Article Featured Left News Politics
la BREAKING: Ex-Cop Father and Son Arrested and Charged With Murder of Black Man Jogging In Neighborhood…President Trump Responds [VIDEO] By 100percentfedup.com Published On :: Fri, 08 May 2020 02:55:40 +0000 The following article, BREAKING: Ex-Cop Father and Son Arrested and Charged With Murder of Black Man Jogging In Neighborhood…President Trump Responds [VIDEO], was first published on 100PercentFedUp.com. The Georgia Bureau of Investigations has arrested a father and son duo, 64-year-old ex-cop, Gregory McMichael, and his son, 34-year-old Travis McMichael for the February murder of 24-year-old Ahmaud Arbery, a black man who was jogging through their neighborhood when they jumped in their truck and pursued him. Yashar Ali shared the news of the […] Continue reading: BREAKING: Ex-Cop Father and Son Arrested and Charged With Murder of Black Man Jogging In Neighborhood…President Trump Responds [VIDEO] ... Full Article Featured
la BREAKING: New Docs Prove Obama Knew Details Of Flynn Wiretapping…Newly Surfaced Video Shows Obama Explaining How He Stays Out Of FBI Investigations By 100percentfedup.com Published On :: Fri, 08 May 2020 16:25:07 +0000 The following article, BREAKING: New Docs Prove Obama Knew Details Of Flynn Wiretapping…Newly Surfaced Video Shows Obama Explaining How He Stays Out Of FBI Investigations, was first published on 100PercentFedUp.com. Barack Obama knew. Documents released yesterday that were used to exonerate President Trump’s new NSA General Flynn, prove that President Barack Obama was aware of the details of Michael Flynn’s intercepted phone calls on December 16 with then-Russian Ambassador Sergey Kislyak. On January 5, 2017, then-Deputy Attorney General, Sally Yates attended an Oval Office meeting […] Continue reading: BREAKING: New Docs Prove Obama Knew Details Of Flynn Wiretapping…Newly Surfaced Video Shows Obama Explaining How He Stays Out Of FBI Investigations ... Full Article Featured Left News Political Correctness
la Greg Gutfeld Levels Ilhan Omar With Epic Response To Her Claim That “White Privilege” Is Reason Charges Were Dropped Against General Flynn By 100percentfedup.com Published On :: Fri, 08 May 2020 19:54:46 +0000 The following article, Greg Gutfeld Levels Ilhan Omar With Epic Response To Her Claim That “White Privilege” Is Reason Charges Were Dropped Against General Flynn, was first published on 100PercentFedUp.com. Yesterday, after 3 1/2 years of having his character and integrity called into question, President Trump's first NSA, General Michael Flynn was finally... Continue reading: Greg Gutfeld Levels Ilhan Omar With Epic Response To Her Claim That “White Privilege” Is Reason Charges Were Dropped Against General Flynn ... Full Article Featured Politics
la Busted! Late-Night Hack Comedian, Jimmy Kimmel Is Forced To Apologize For Sharing Highly Edited Video Of VP Pence To Make Him Look Bad By 100percentfedup.com Published On :: Sat, 09 May 2020 01:45:58 +0000 The following article, Busted! Late-Night Hack Comedian, Jimmy Kimmel Is Forced To Apologize For Sharing Highly Edited Video Of VP Pence To Make Him Look Bad, was first published on 100PercentFedUp.com. Last night, Jimmy Kimmel, host of the low-rated, late-night Jimmy Kimmel Show, shared a deceptively edited video clip of Vice President Pence delivering PPE to a nursing home. Today, liberal activist Matt McDermott tweeted the videotaped segment on VP Pence that was edited to make the vice president look like he was faking a delivery […] Continue reading: Busted! Late-Night Hack Comedian, Jimmy Kimmel Is Forced To Apologize For Sharing Highly Edited Video Of VP Pence To Make Him Look Bad ... Full Article Featured Politics
la Video: Dallas Salon Owner Shelley Luther Gets Surprise Visit from Sen Ted Cruz for a Celebratory Haircut By 100percentfedup.com Published On :: Sat, 09 May 2020 13:19:16 +0000 The following article, Video: Dallas Salon Owner Shelley Luther Gets Surprise Visit from Sen Ted Cruz for a Celebratory Haircut, was first published on 100PercentFedUp.com. When Dallas, Texas salon owner Shelley Luther opened her salon in defiance of the lockdown order in Texas, she was visited numerous times by the local police and then sentenced to 7 days in jail with a fine of $7,000. After public outrage at her punishment, the Texas Supreme Court stepped in to demand her […] Continue reading: Video: Dallas Salon Owner Shelley Luther Gets Surprise Visit from Sen Ted Cruz for a Celebratory Haircut ... Full Article Breaking Featured Government News Politics
la Council votes against proposed cannabis store location in Lakeshore By windsor.ctvnews.ca Published On :: Fri, 8 May 2020 16:54:00 -0400 In Lakeshore, it may be a little while longer before a retail cannabis store opens. Full Article
la Windsor Assembly Plant on track for May 19 restart By windsor.ctvnews.ca Published On :: Fri, 8 May 2020 16:21:00 -0400 The Fiat-Chrysler Windsor Assembly Plant looks to be on track for a May 19 reopening according to union leadership. Full Article
la Owners of new retail cannabis store hoping to open soon in Pillette Village By windsor.ctvnews.ca Published On :: Fri, 8 May 2020 16:51:00 -0400 At a time when store front vacancies are growing thanks to COVID-19, a new retail cannabis store in Pillette Village is hoping to open soon. Full Article
la COVID-19 outbreak at Health Sciences Centre declared over By winnipeg.ctvnews.ca Published On :: Fri, 8 May 2020 13:34:00 -0600 Health officials with the Manitoba government announced on Friday that a COVID-19 outbreak at the Health Sciences Centre has officially been declared over. Full Article
la Little Richard, flamboyant rock 'n' roll pioneer, dead at 87 By www.ctvnews.ca Published On :: Sat, 9 May 2020 08:55:00 -0600 Little Richard, the self-proclaimed "architect of rock 'n' roll" whose piercing wail, pounding piano and towering pompadour irrevocably altered popular music while introducing black R&B to white America, has died Saturday. He was 87. Full Article
la Michela Gariboldi By feedproxy.google.com Published On :: Thu, 08 Aug 2019 15:35:42 +0000 Research Assistant, Global Economy and Finance Programme Full Article
la New Dimensions in Trade Law By feedproxy.google.com Published On :: Tue, 01 Oct 2019 10:25:01 +0000 Research Event 6 November 2019 - 9:15am to 4:15pm Chatham House | 10 St James's Square | London | SW1Y 4LE Event participants Speakers include:Dr Lorand Bartels, Reader in International Law; Fellow, Trinity House, University of CambridgeLaura Bannister, Senior Adviser on EU-UK Trade, Trade Justice MovementPeter Holmes, Fellow, UKTPO; Reader in Economics, University of SussexAndrew Hood, Partner, Regulatory & Trade, FieldFisher LLP At this event, which forms the second annual UK Trade Policy Observatory conference, there will be six presentations over the course of the day before concluding with a panel discussion and Q&A. This year’s conference will focus on the following legal areas of trade policy:Blockchain: Creating and Eliminating Trade in ServicesChina's Role in the International Trading SystemOfficial Export Support: Compliance and Competition ConcernsStrategic Litigation and Health Regulation: Implications for International Economic LawDevelopment, Labour Standards and Sustainability in Trade AgreementsRetaining Versus Reforming EU Food Safety Legislation: Selected Issues for a US-UK Trade NegotiationTo register for this event, please click here. Department/project Global Economy and Finance Programme, UK Trade Policy Observatory Full Article
la The EU Cannot Build a Foreign Policy on Regulatory Power Alone By feedproxy.google.com Published On :: Tue, 11 Feb 2020 16:33:26 +0000 11 February 2020 Alan Beattie Associate Fellow, Global Economy and Finance Programme and Europe Programme @alanbeattie LinkedIn Brussels will find its much-vaunted heft in setting standards cannot help it advance its geopolitical interests. 2020-02-11-Leyen.jpg EU Commission President Ursula von der Leyen speaks at the European Parliament in Strasbourg in February. Photo: Getty Images. There are two well-established ideas in trade. Individually, they are correct. Combined, they can lead to a conclusion that is unfortunately wrong.The first idea is that, across a range of economic sectors, the EU and the US have been engaged in a battle to have their model of regulation accepted as the global one, and that the EU is generally winning.The second is that governments can use their regulatory power to extend strategic and foreign policy influence.The conclusion would seem to be that the EU, which has for decades tried to develop a foreign policy, should be able to use its superpower status in regulation and trade to project its interests and its values abroad.That’s the theory. It’s a proposition much welcomed by EU policymakers, who know they are highly unlikely any time soon to acquire any of the tools usually required to run an effective foreign policy.The EU doesn’t have an army it can send into a shooting war, enough military or political aid to prop up or dispense of governments abroad, or a centralized intelligence service. Commission President Ursula von der Leyen has declared her outfit to be a ‘geopolitical commission’, and is casting about for any means of making that real.Through the ‘Brussels effect’ whereby European rules and standards are exported via both companies and governments, the EU has indeed won many regulatory battles with the US.Its cars, chemicals and product safety regulations are more widely adopted round the world than their American counterparts. In the absence of any coherent US offering, bar some varied state-level systems, the General Data Protection Regulation (GDPR) is the closest thing the world has to a single model for data privacy, and variants of it are being adopted by dozens of countries.The problem is this. Those parts of global economic governance where the US is dominant – particularly the dollar payments system – are highly conducive to projecting US power abroad. The extraterritorial reach of secondary sanctions, plus the widespread reliance of banks and companies worldwide on dollar funding – and hence the American financial system – means that the US can precisely target its influence.The EU can enforce trade sanctions, but not in such a powerful and discriminatory way, and it will always be outgunned by the US. Donald Trump could in effect force European companies to join in his sanctions on Iran when he pulled out of the nuclear deal, despite EU legislation designed to prevent their businesses being bullied. He can go after the chief financial officer of Huawei for allegedly breaching those sanctions.By contrast, the widespread adoption of GDPR or data protection regimes inspired by it may give the EU a warm glow of satisfaction, but it cannot be turned into a geopolitical tool in the same way.Nor, necessarily, does it particularly benefit the EU economy. Europe’s undersized tech sector seems unlikely to unduly benefit from the fact that data protection rules were written in the EU. Indeed, one common criticism of the regulations is that they entrench the power of incumbent tech giants like Google.There is a similar pattern at work in the adoption of new technologies such as artificial intelligence and the Internet of Things. In that field, the EU and its member states are also facing determined competition from China, which has been pushing its technologies and standards through forums such as the International Telecommunication Union.The EU has been attempting to write international rules for the use of AI which it hopes to be widely adopted. But again, these are a constraint on the use of new technologies largely developed by others, not the control of innovation.By contrast, China has created a vast domestic market in technologies like facial recognition and unleashed its own companies on it. The resulting surveillance kit can then be marketed to emerging market governments as part of China’s enduring foreign policy campaign to build up supporters in the developing world.If it genuinely wants to turn its economic power into geopolitical influence – and it’s not entirely clear what it would do with it if it did – the EU needs to recognize that not all forms of regulatory and trading dominance are the same.Providing public goods to the world economy is all very well. But unless they are so particular in nature that they project uniquely European values and interests, that makes the EU a supplier of useful plumbing but not a global architect of power.On the other hand, it could content itself with its position for the moment. It could recognize that not until enough hard power – guns, intelligence, money – is transferred from the member states to the centre, or until the member states start acting collectively, will the EU genuinely become a geopolitical force. Speaking loudly and carrying a stick of foam rubber is rarely a way to gain credibility in international relations.This article is part of a series of publications and roundtable discussions in the Chatham House Global Trade Policy Forum. Full Article
la A comprehensive evaluation of a typical plant telomeric G-quadruplex (G4) DNA reveals the dynamics of G4 formation, rearrangement, and unfolding [Plant Biology] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 Telomeres are specific nucleoprotein structures that are located at the ends of linear eukaryotic chromosomes and play crucial roles in genomic stability. Telomere DNA consists of simple repeats of a short G-rich sequence: TTAGGG in mammals and TTTAGGG in most plants. In recent years, the mammalian telomeric G-rich repeats have been shown to form G-quadruplex (G4) structures, which are crucial for modulating telomere functions. Surprisingly, even though plant telomeres are essential for plant growth, development, and environmental adaptions, only few reports exist on plant telomeric G4 DNA (pTG4). Here, using bulk and single-molecule assays, including CD spectroscopy, and single-molecule FRET approaches, we comprehensively characterized the structure and dynamics of a typical plant telomeric sequence, d[GGG(TTTAGGG)3]. We found that this sequence can fold into mixed G4s in potassium, including parallel and antiparallel structures. We also directly detected intermediate dynamic transitions, including G-hairpin, parallel G-triplex, and antiparallel G-triplex structures. Moreover, we observed that pTG4 is unfolded by the AtRecQ2 helicase but not by AtRecQ3. The results of our work shed light on our understanding about the existence, topological structures, stability, intermediates, unwinding, and functions of pTG4. Full Article
la Spectral and photochemical diversity of tandem cysteine cyanobacterial phytochromes [Plant Biology] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 The atypical trichromatic cyanobacterial phytochrome NpTP1 from Nostoc punctiforme ATCC 29133 is a linear tetrapyrrole (bilin)-binding photoreceptor protein that possesses tandem-cysteine residues responsible for shifting its light-sensing maximum to the violet spectral region. Using bioinformatics and phylogenetic analyses, here we established that tandem-cysteine cyanobacterial phytochromes (TCCPs) compose a well-supported monophyletic phytochrome lineage distinct from prototypical red/far-red cyanobacterial phytochromes. To investigate the light-sensing diversity of this family, we compared the spectroscopic properties of NpTP1 (here renamed NpTCCP) with those of three phylogenetically diverged TCCPs identified in the draft genomes of Tolypothrix sp. PCC7910, Scytonema sp. PCC10023, and Gloeocapsa sp. PCC7513. Recombinant photosensory core modules of ToTCCP, ScTCCP, and GlTCCP exhibited violet-blue–absorbing dark-states consistent with dual thioether-linked phycocyanobilin (PCB) chromophores. Photoexcitation generated singly-linked photoproduct mixtures with variable ratios of yellow-orange and red-absorbing species. The photoproduct ratio was strongly influenced by pH and by mutagenesis of TCCP- and phytochrome-specific signature residues. Our experiments support the conclusion that both photoproduct species possess protonated 15E bilin chromophores, but differ in the ionization state of the noncanonical “second” cysteine sulfhydryl group. We found that the ionization state of this and other residues influences subsequent conformational change and downstream signal transmission. We also show that tandem-cysteine phytochromes present in eukaryotes possess similar amino acid substitutions within their chromophore-binding pocket, which tune their spectral properties in an analogous fashion. Taken together, our findings provide a roadmap for tailoring the wavelength specificity of plant phytochromes to optimize plant performance in diverse natural and artificial light environments. Full Article
la Repression of sphingosine kinase (SK)-interacting protein (SKIP) in acute myeloid leukemia diminishes SK activity and its re-expression restores SK function [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 Previous studies have shown that sphingosine kinase interacting protein (SKIP) inhibits sphingosine kinase (SK) function in fibroblasts. SK phosphorylates sphingosine producing the potent signaling molecule sphingosine-1-phosphate (S1P). SKIP gene (SPHKAP) expression is silenced by hypermethylation of its promoter in acute myeloid leukemia (AML). However, why SKIP activity is silenced in primary AML cells is unclear. Here, we investigated the consequences of SKIP down-regulation in AML primary cells and the effects of SKIP re-expression in leukemic cell lines. Using targeted ultra-HPLC-tandem MS (UPLC-MS/MS), we measured sphingolipids (including S1P and ceramides) in AML and control cells. Primary AML cells had significantly lower SK activity and intracellular S1P concentrations than control cells, and SKIP-transfected leukemia cell lines exhibited increased SK activity. These findings show that SKIP re-expression enhances SK activity in leukemia cells. Furthermore, other bioactive sphingolipids such as ceramide were also down-regulated in primary AML cells. Of note, SKIP re-expression in leukemia cells increased ceramide levels 2-fold, inactivated the key signaling protein extracellular signal-regulated kinase, and increased apoptosis following serum deprivation or chemotherapy. These results indicate that SKIP down-regulation in AML reduces SK activity and ceramide levels, an effect that ultimately inhibits apoptosis in leukemia cells. The findings of our study contrast with previous results indicating that SKIP inhibits SK function in fibroblasts and therefore challenge the notion that SKIP always inhibits SK activity. Full Article
la The FKH domain in FOXP3 mRNA frequently contains mutations in hepatocellular carcinoma that influence the subcellular localization and functions of FOXP3 [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 The transcription factor forkhead box P3 (FOXP3) is a biomarker for regulatory T cells and can also be expressed in cancer cells, but its function in cancer appears to be divergent. The role of hepatocyte-expressed FOXP3 in hepatocellular carcinoma (HCC) is unknown. Here, we collected tumor samples and clinical information from 115 HCC patients and used five human cancer cell lines. We examined FOXP3 mRNA sequences for mutations, used a luciferase assay to assess promoter activities of FOXP3's target genes, and employed mouse tumor models to confirm in vitro results. We detected mutations in the FKH domain of FOXP3 mRNAs in 33% of the HCC tumor tissues, but in none of the adjacent nontumor tissues. None of the mutations occurred at high frequency, indicating that they occurred randomly. Notably, the mutations were not detected in the corresponding regions of FOXP3 genomic DNA, and many of them resulted in amino acid substitutions in the FKH region, altering FOXP3's subcellular localization. FOXP3 delocalization from the nucleus to the cytoplasm caused loss of transcriptional regulation of its target genes, inactivated its tumor-inhibitory capability, and changed cellular responses to histone deacetylase (HDAC) inhibitors. More complex FKH mutations appeared to be associated with worse prognosis in HCC patients. We conclude that mutations in the FKH domain of FOXP3 mRNA frequently occur in HCC and that these mutations are caused by errors in transcription and are not derived from genomic DNA mutations. Our results suggest that transcriptional mutagenesis of FOXP3 plays a role in HCC. Full Article
la Inhibition of the erythropoietin-producing receptor EPHB4 antagonizes androgen receptor overexpression and reduces enzalutamide resistance [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 Prostate cancer (PCa) cells heavily rely on an active androgen receptor (AR) pathway for their survival. Enzalutamide (MDV3100) is a second-generation antiandrogenic drug that was approved by the Food and Drug Administration in 2012 to treat patients with castration-resistant prostate cancer (CRPC). However, emergence of resistance against this drug is inevitable, and it has been a major challenge to develop interventions that help manage enzalutamide-resistant CRPC. Erythropoietin-producing human hepatocellular (Eph) receptors are targeted by ephrin protein ligands and have a broad range of functions. Increasing evidence indicates that this signaling pathway plays an important role in tumorigenesis. Overexpression of EPH receptor B4 (EPHB4) has been observed in multiple types of cancer, being closely associated with proliferation, invasion, and metastasis of tumors. Here, using RNA-Seq analyses of clinical and preclinical samples, along with several biochemical and molecular methods, we report that enzalutamide-resistant PCa requires an active EPHB4 pathway that supports drug resistance of this tumor type. Using a small kinase inhibitor and RNAi-based gene silencing to disrupt EPHB4 activity, we found that these disruptions re-sensitize enzalutamide-resistant PCa to the drug both in vitro and in vivo. Mechanistically, we found that EPHB4 stimulates the AR by inducing proto-oncogene c-Myc (c-Myc) expression. Taken together, these results provide critical insight into the mechanism of enzalutamide resistance in PCa, potentially offering a therapeutic avenue for enhancing the efficacy of enzalutamide to better manage this common malignancy. Full Article
la A peroxisome deficiency-induced reductive cytosol state up-regulates the brain-derived neurotrophic factor pathway [Metabolism] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 The peroxisome is a subcellular organelle that functions in essential metabolic pathways, including biosynthesis of plasmalogens, fatty acid β-oxidation of very-long-chain fatty acids, and degradation of hydrogen peroxide. Peroxisome biogenesis disorders (PBDs) manifest as severe dysfunction in multiple organs, including the central nervous system (CNS), but the pathogenic mechanisms in PBDs are largely unknown. Because CNS integrity is coordinately established and maintained by neural cell interactions, we here investigated whether cell-cell communication is impaired and responsible for the neurological defects associated with PBDs. Results from a noncontact co-culture system consisting of primary hippocampal neurons with glial cells revealed that a peroxisome-deficient astrocytic cell line secretes increased levels of brain-derived neurotrophic factor (BDNF), resulting in axonal branching of the neurons. Of note, the BDNF expression in astrocytes was not affected by defects in plasmalogen biosynthesis and peroxisomal fatty acid β-oxidation in the astrocytes. Instead, we found that cytosolic reductive states caused by a mislocalized catalase in the peroxisome-deficient cells induce the elevation in BDNF secretion. Our results suggest that peroxisome deficiency dysregulates neuronal axogenesis by causing a cytosolic reductive state in astrocytes. We conclude that astrocytic peroxisomes regulate BDNF expression and thereby support neuronal integrity and function. Full Article
la Heterotrimeric Gq proteins as therapeutic targets? [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 Heterotrimeric G proteins are the core upstream elements that transduce and amplify the cellular signals from G protein–coupled receptors (GPCRs) to intracellular effectors. GPCRs are the largest family of membrane proteins encoded in the human genome and are the targets of about one-third of prescription medicines. However, to date, no single therapeutic agent exerts its effects via perturbing heterotrimeric G protein function, despite a plethora of evidence linking G protein malfunction to human disease. Several recent studies have brought to light that the Gq family–specific inhibitor FR900359 (FR) is unexpectedly efficacious in silencing the signaling of Gq oncoproteins, mutant Gq variants that mostly exist in the active state. These data not only raise the hope that researchers working in drug discovery may be able to potentially strike Gq oncoproteins from the list of undruggable targets, but also raise questions as to how FR achieves its therapeutic effect. Here, we place emphasis on these recent studies and explain why they expand our pharmacological armamentarium for targeting Gq protein oncogenes as well as broaden our mechanistic understanding of Gq protein oncogene function. We also highlight how this novel insight impacts the significance and utility of using G(q) proteins as targets in drug discovery efforts. Full Article
la N{alpha}-Acetylation of the virulence factor EsxA is required for mycobacterial cytosolic translocation and virulence [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-04-24T06:08:45-07:00 The Mycobacterium tuberculosis virulence factor EsxA and its chaperone EsxB are secreted as a heterodimer (EsxA:B) and are crucial for mycobacterial escape from phagosomes and cytosolic translocation. Current findings support the idea that for EsxA to interact with host membranes, EsxA must dissociate from EsxB at low pH. However, the molecular mechanism by which the EsxA:B heterodimer separates is not clear. In the present study, using liposome-leakage and cytotoxicity assays, LC-MS/MS–based proteomics, and CCF-4 FRET analysis, we obtained evidence that the Nα-acetylation of the Thr-2 residue on EsxA, a post-translational modification that is present in mycobacteria but absent in Escherichia coli, is required for the EsxA:B separation. Substitutions at Thr-2 that precluded Nα-acetylation inhibited the heterodimer separation and hence prevented EsxA from interacting with the host membrane, resulting in attenuated mycobacterial cytosolic translocation and virulence. Molecular dynamics simulations revealed that at low pH, the Nα-acetylated Thr-2 makes direct and frequent “bind-and-release” contacts with EsxB, which generates a force that pulls EsxB away from EsxA. In summary, our findings provide evidence that the Nα-acetylation at Thr-2 of EsxA facilitates dissociation of the EsxA:B heterodimer required for EsxA membrane permeabilization and mycobacterial cytosolic translocation and virulence. Full Article
la ER stress increases store-operated Ca2+ entry (SOCE) and augments basal insulin secretion in pancreatic beta cells [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-04-24T06:08:45-07:00 Type 2 diabetes mellitus (T2DM) is characterized by impaired glucose-stimulated insulin secretion and increased peripheral insulin resistance. Unremitting endoplasmic reticulum (ER) stress can lead to beta-cell apoptosis and has been linked to type 2 diabetes. Although many studies have attempted to link ER stress and T2DM, the specific effects of ER stress on beta-cell function remain incompletely understood. To determine the interrelationship between ER stress and beta-cell function, here we treated insulin-secreting INS-1(832/13) cells or isolated mouse islets with the ER stress–inducer tunicamycin (TM). TM induced ER stress as expected, as evidenced by activation of the unfolded protein response. Beta cells treated with TM also exhibited concomitant alterations in their electrical activity and cytosolic free Ca2+ oscillations. As ER stress is known to reduce ER Ca2+ levels, we tested the hypothesis that the observed increase in Ca2+ oscillations occurred because of reduced ER Ca2+ levels and, in turn, increased store-operated Ca2+ entry. TM-induced cytosolic Ca2+ and membrane electrical oscillations were acutely inhibited by YM58483, which blocks store-operated Ca2+ channels. Significantly, TM-treated cells secreted increased insulin under conditions normally associated with only minimal release, e.g. 5 mm glucose, and YM58483 blocked this secretion. Taken together, these results support a critical role for ER Ca2+ depletion–activated Ca2+ current in mediating Ca2+-induced insulin secretion in response to ER stress. Full Article
la Mechanistic insights explain the transforming potential of the T507K substitution in the protein-tyrosine phosphatase SHP2 [Signal Transduction] By feedproxy.google.com Published On :: 2020-05-01T00:06:09-07:00 The protein-tyrosine phosphatase SHP2 is an allosteric enzyme critical for cellular events downstream of growth factor receptors. Mutations in the SHP2 gene have been linked to many different types of human diseases, including developmental disorders, leukemia, and solid tumors. Unlike most SHP2-activating mutations, the T507K substitution in SHP2 is unique in that it exhibits oncogenic Ras-like transforming activity. However, the biochemical basis of how the SHP2/T507K variant elicits transformation remains unclear. By combining kinetic and biophysical methods, X-ray crystallography, and molecular modeling, as well as using cell biology approaches, here we uncovered that the T507K substitution alters both SHP2 substrate specificity and its allosteric regulatory mechanism. We found that although SHP2/T507K exists in the closed, autoinhibited conformation similar to the WT enzyme, the interactions between its N-SH2 and protein-tyrosine phosphatase domains are weakened such that SHP2/T507K possesses a higher affinity for the scaffolding protein Grb2-associated binding protein 1 (Gab1). We also discovered that the T507K substitution alters the structure of the SHP2 active site, resulting in a change in SHP2 substrate preference for Sprouty1, a known negative regulator of Ras signaling and a potential tumor suppressor. Our results suggest that SHP2/T507K's shift in substrate specificity coupled with its preferential association of SHP2/T507K with Gab1 enable the mutant SHP2 to more efficiently dephosphorylate Sprouty1 at pTyr-53. This dephosphorylation hyperactivates Ras signaling, which is likely responsible for SHP2/T507K's Ras-like transforming activity. Full Article
la Modification of a PE/PPE substrate pair reroutes an Esx substrate pair from the mycobacterial ESX-1 type VII secretion system to the ESX-5 system [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-05-01T00:06:09-07:00 Bacterial type VII secretion systems secrete a wide range of extracellular proteins that play important roles in bacterial viability and in interactions of pathogenic mycobacteria with their hosts. Mycobacterial type VII secretion systems consist of five subtypes, ESX-1–5, and have four substrate classes, namely, Esx, PE, PPE, and Esp proteins. At least some of these substrates are secreted as heterodimers. Each ESX system mediates the secretion of a specific set of Esx, PE, and PPE proteins, raising the question of how these substrates are recognized in a system-specific fashion. For the PE/PPE heterodimers, it has been shown that they interact with their cognate EspG chaperone and that this chaperone determines the designated secretion pathway. However, both structural and pulldown analyses have suggested that EspG cannot interact with the Esx proteins. Therefore, the determining factor for system specificity of the Esx proteins remains unknown. Here, we investigated the secretion specificity of the ESX-1 substrate pair EsxB_1/EsxA_1 in Mycobacterium marinum. Although this substrate pair was hardly secreted when homologously expressed, it was secreted when co-expressed together with the PE35/PPE68_1 pair, indicating that this pair could stimulate secretion of the EsxB_1/EsxA_1 pair. Surprisingly, co-expression of EsxB_1/EsxA_1 with a modified PE35/PPE68_1 version that carried the EspG5 chaperone-binding domain, previously shown to redirect this substrate pair to the ESX-5 system, also resulted in redirection and co-secretion of the Esx pair via ESX-5. Our results suggest a secretion model in which PE35/PPE68_1 determines the system-specific secretion of EsxB_1/EsxA_1. Full Article
la Structure-based discovery of a small-molecule inhibitor of methicillin-resistant Staphylococcus aureus virulence [Molecular Biophysics] By feedproxy.google.com Published On :: 2020-05-01T00:06:09-07:00 The rapid emergence and dissemination of methicillin-resistant Staphylococcus aureus (MRSA) strains poses a major threat to public health. MRSA possesses an arsenal of secreted host-damaging virulence factors that mediate pathogenicity and blunt immune defenses. Panton–Valentine leukocidin (PVL) and α-toxin are exotoxins that create lytic pores in the host cell membrane. They are recognized as being important for the development of invasive MRSA infections and are thus potential targets for antivirulence therapies. Here, we report the high-resolution X-ray crystal structures of both PVL and α-toxin in their soluble, monomeric, and oligomeric membrane-inserted pore states in complex with n-tetradecylphosphocholine (C14PC). The structures revealed two evolutionarily conserved phosphatidylcholine-binding mechanisms and their roles in modulating host cell attachment, oligomer assembly, and membrane perforation. Moreover, we demonstrate that the soluble C14PC compound protects primary human immune cells in vitro against cytolysis by PVL and α-toxin and hence may serve as the basis for the development of an antivirulence agent for managing MRSA infections. Full Article
la {alpha}-Synuclein filaments from transgenic mouse and human synucleinopathy-containing brains are maȷor seed-competent species [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 Assembled α-synuclein in nerve cells and glial cells is the defining pathological feature of neurodegenerative diseases called synucleinopathies. Seeds of α-synuclein can induce the assembly of monomeric protein. Here, we used sucrose gradient centrifugation and transiently transfected HEK 293T cells to identify the species of α-synuclein from the brains of homozygous, symptomatic mice transgenic for human mutant A53T α-synuclein (line M83) that seed aggregation. The most potent fractions contained Sarkosyl-insoluble assemblies enriched in filaments. We also analyzed six cases of idiopathic Parkinson's disease (PD), one case of familial PD, and six cases of multiple system atrophy (MSA) for their ability to induce α-synuclein aggregation. The MSA samples were more potent than those of idiopathic PD in seeding aggregation. We found that following sucrose gradient centrifugation, the most seed-competent fractions from PD and MSA brains are those that contain Sarkosyl-insoluble α-synuclein. The fractions differed between PD and MSA, consistent with the presence of distinct conformers of assembled α-synuclein in these different samples. We conclude that α-synuclein filaments are the main driving force for amplification and propagation of pathology in synucleinopathies. Full Article
la Non-photopic and photopic visual cycles differentially regulate immediate, early, and late phases of cone photoreceptor-mediated vision [Molecular Bases of Disease] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 Cone photoreceptors in the retina enable vision over a wide range of light intensities. However, the processes enabling cone vision in bright light (i.e. photopic vision) are not adequately understood. Chromophore regeneration of cone photopigments may require the retinal pigment epithelium (RPE) and/or retinal Müller glia. In the RPE, isomerization of all-trans-retinyl esters to 11-cis-retinol is mediated by the retinoid isomerohydrolase Rpe65. A putative alternative retinoid isomerase, dihydroceramide desaturase-1 (DES1), is expressed in RPE and Müller cells. The retinol-isomerase activities of Rpe65 and Des1 are inhibited by emixustat and fenretinide, respectively. Here, we tested the effects of these visual cycle inhibitors on immediate, early, and late phases of cone photopic vision. In zebrafish larvae raised under cyclic light conditions, fenretinide impaired late cone photopic vision, while the emixustat-treated zebrafish unexpectedly had normal vision. In contrast, emixustat-treated larvae raised under extensive dark-adaptation displayed significantly attenuated immediate photopic vision concomitant with significantly reduced 11-cis-retinaldehyde (11cRAL). Following 30 min of light, early photopic vision was recovered, despite 11cRAL levels remaining significantly reduced. Defects in immediate cone photopic vision were rescued in emixustat- or fenretinide-treated larvae following exogenous 9-cis-retinaldehyde supplementation. Genetic knockout of Des1 (degs1) or retinaldehyde-binding protein 1b (rlbp1b) did not eliminate photopic vision in zebrafish. Our findings define molecular and temporal requirements of the nonphotopic or photopic visual cycles for mediating vision in bright light. Full Article
la Pro-515 of the dynamin-like GTPase MxB contributes to HIV-1 inhibition by regulating MxB oligomerization and binding to HIV-1 capsid [Microbiology] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 Interferon-regulated myxovirus resistance protein B (MxB) is an interferon-induced GTPase belonging to the dynamin superfamily. It inhibits infection with a wide range of different viruses, including HIV-1, by impairing viral DNA entry into the nucleus. Unlike the related antiviral GTPase MxA, MxB possesses an N-terminal region that contains a nuclear localization signal and is crucial for inhibiting HIV-1. Because MxB previously has been shown to reside in both the nuclear envelope and the cytoplasm, here we used bioinformatics and biochemical approaches to identify a nuclear export signal (NES) responsible for MxB's cytoplasmic location. Using the online computational tool LocNES (Locating Nuclear Export Signals or NESs), we identified five putative NES candidates in MxB and investigated whether their deletion caused nuclear localization of MxB. Our results revealed that none of the five deletion variants relocates to the nucleus, suggesting that these five predicted NES sequences do not confer NES activity. Interestingly, deletion of one sequence, encompassing amino acids 505–527, abrogated the anti-HIV-1 activity of MxB. Further mutation experiments disclosed that amino acids 515–519, and Pro-515 in particular, regulate MxB oligomerization and its binding to HIV-1 capsid, thereby playing an important role in MxB-mediated restriction of HIV-1 infection. In summary, our results indicate that none of the five predicted NES sequences in MxB appears to be required for its nuclear export. Our findings also reveal several residues in MxB, including Pro-515, critical for its oligomerization and anti-HIV-1 function. Full Article
la A neuroglobin-based high-affinity ligand trap reverses carbon monoxide-induced mitochondrial poisoning [Molecular Biophysics] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 Carbon monoxide (CO) remains the most common cause of human poisoning. The consequences of CO poisoning include cardiac dysfunction, brain injury, and death. CO causes toxicity by binding to hemoglobin and by inhibiting mitochondrial cytochrome c oxidase (CcO), thereby decreasing oxygen delivery and inhibiting oxidative phosphorylation. We have recently developed a CO antidote based on human neuroglobin (Ngb-H64Q-CCC). This molecule enhances clearance of CO from red blood cells in vitro and in vivo. Herein, we tested whether Ngb-H64Q-CCC can also scavenge CO from CcO and attenuate CO-induced inhibition of mitochondrial respiration. Heart tissue from mice exposed to 3% CO exhibited a 42 ± 19% reduction in tissue respiration rate and a 33 ± 38% reduction in CcO activity compared with unexposed mice. Intravenous infusion of Ngb-H64Q-CCC restored respiration rates to that of control mice correlating with higher electron transport chain CcO activity in Ngb-H64Q-CCC–treated compared with PBS-treated, CO-poisoned mice. Further, using a Clark-type oxygen electrode, we measured isolated rat liver mitochondrial respiration in the presence and absence of saturating solutions of CO (160 μm) and nitric oxide (100 μm). Both CO and NO inhibited respiration, and treatment with Ngb-H64Q-CCC (100 and 50 μm, respectively) significantly reversed this inhibition. These results suggest that Ngb-H64Q-CCC mitigates CO toxicity by scavenging CO from carboxyhemoglobin, improving systemic oxygen delivery and reversing the inhibitory effects of CO on mitochondria. We conclude that Ngb-H64Q-CCC or other CO scavengers demonstrate potential as antidotes that reverse the clinical and molecular effects of CO poisoning. Full Article
