ino Illinois' Alan Griffin gets suspension for stepping on Purdue's Sasha Stefanovic By rssfeeds.indystar.com Published On :: Thu, 23 Jan 2020 22:27:36 +0000 First-half incident resulted in an ejection for Illini player as he stepped on Boilermakers guard Full Article
ino Bargersville police officer involved in Friday crash sustains minor injury By rssfeeds.indystar.com Published On :: Sat, 04 Apr 2020 16:59:24 +0000 A Bargersville police officer and the driver of an SUV were taken to the hospital with minor injuries after a vehicle crash Friday. Full Article
ino Flash floods hit parts of Gorseinon and Carmarthen By www.bbc.co.uk Published On :: Sat, 09 May 2020 16:45:41 GMT Properties have been flooded after thunderstorms hit parts of south and west Wales on Saturday. Full Article
ino Timeline: San Marino By news.bbc.co.uk Published On :: Tue, 08 Nov 2011 12:28:21 GMT A chronology of key events Full Article Country profiles
ino Country profile: San Marino By news.bbc.co.uk Published On :: Tue, 08 Nov 2011 12:26:49 GMT An overview of San Marino including key facts, political leaders and notes on the media Full Article Country profiles
ino Kabardino-Balkaria profile By news.bbc.co.uk Published On :: Tue, 22 Nov 2011 15:08:38 GMT A profile of the southern Russian republic Full Article Country profiles
ino Domino’s will start delivering pizzas via an autonomous robot this fall By www.washingtonpost.com Published On :: Mon, 17 Jun 2019 19:53:05 +0000 Domino's plans to start delivering pizzas using an autonomous, unmanned vehicle. The announcement is the latest example of technology companies using robots to deliver food. Full Article
ino Google’s balloon project has a new test: Providing Internet access to ‘mountainous villagers’ in Kenya By www.washingtonpost.com Published On :: Fri, 05 Jul 2019 20:43:31 +0000 Loon — an Internet-providing balloon service owned by Alphabet, Google’s parent company — will give “mountain villagers” in Kenya the opportunity to purchase 4G service. Full Article
ino Soccer star Megan Rapinoe will be secretary of state if Jay Inslee has any say By www.washingtonpost.com Published On :: Sun, 14 Jul 2019 17:37:18 +0000 The presidential candidate and Washington governor wants Rapinoe in his administration — should he get elected. Full Article
ino Amino Showcases New Live Media Gateway at TelcoTV 2012 By www.tvover.net Published On :: Tue, 16 Oct 2012 21:02:39 GMT Amino Communications will showcase at TelcoTV 2012 in Las Vegas a new breakthrough version of its Live media gateway that delivers a powerful combination of pay-TV security and exciting Android-based OTT content delivery. The Live media gateway is the first hybrid/OTT media gateway to be powered by the new Intel Atom CE 5300 media processor. This combines dual core processing power with a range of new enhanced transcoding and encoding capabilities with multi-screen distribution to deliver content seamlessly to TVs, smartphones and tablets around the home. Now the product has been enhanced – with a secure Linux environment, based on Amino’s proven Aminet software stack, for the delivery of payTV video, and a controlled Android virtual machine to support a wide range of OTT applications and services. The device will utilise the unparalleled hardware virtualisation features available in the Intel CE5300 SoC to run multiple virtual machines at full speed. Full Article IP Equipment;IPTV Events
ino The more ominous part of the Trump Sharpie incident By www.washingtonpost.com Published On :: Mon, 09 Sep 2019 22:54:42 +0000 Trump’s attempted manipulations of official metrics degrade our democracy, economy and public safety. Full Article
ino Arduino Cookbook By shop.oreilly.com Published On :: Wed, 06 May 2020 05:00:34 PDT Want to create devices that interact with the physical world? This cookbook is perfect for anyone who wants to experiment with the popular Arduino microcontroller and programming environment. Updated for Arduino 1.8, the third edition includes more than 200 tips and techniques for building a variety of objects and prototypes such as IoT solutions, environmental monitors, location and position-aware systems, and products that can respond to touch, sound, heat, and light. You don’t need to have mastered Arduino or programming to get started. Full Article
ino German rights holders want to sue Kino.to viewers By feedproxy.google.com Published On :: Mon, 13 Jun 2011 22:51:23 -0700 Have you ever watched a TV show episode or Hollywood blockbuster on a website that didnt have the proper licenses? Then you could be in trouble, at least if the example of Kino.to catches on. Rights holders are threatening to sue users of the now-defunct site. Continue reading on NewTeeVee. Tags: kino.to, newteevee Full Article Lawsuits
ino AT#53 - Travel to Chicago, Illinois By traffic.libsyn.com Published On :: Sat, 12 Aug 2006 18:13:00 +0000 Chicago, Illionois Full Article
ino AT#260 - Biking Spain's Camino di Santiago By europe.amateurtraveler.com Published On :: Sat, 25 Dec 2010 15:00:00 +0000 The Amateur Traveler talks to Ben Dixon about biking the route of the Camino di Santiago in northern Spain. The Camino di Santiago is a an ancient pilgrim trail that runs across northern Spain from the border with France to Santiago di Compostella. The cathedral in Santiago di Compostella reputedly holds the remains of St. James. Modern day pilgrims are sometimes drawn by faith and sometimes just drawn by the journey itself to trek for a month across the varied landscapes of northern Spain. As an American, Ben did not have a month of vacation but rather than be one of those pilgrims who only traverses the very end of the trail he chose to take on the pilgrim route on a bicycle. Along the way he stayed in the hostels that house the many pilgrims where he met people who had been walking from far beyond the border with France. Through countryside, cities and picturesque towns he rode in both sunshine and driving rain, but the highlight of the trip might not be what you think. Full Article
ino AT#340 - Travel to the Microstates of Europe (Andorra, Liechtenstein, Monaco, San Marino) By europe.amateurtraveler.com Published On :: Sat, 08 Sep 2012 21:54:00 +0000 The Amateur Traveler talks to to Gary Arndt about the 4 micro-states in Europe: Andorra, Liechtenstein, Monaco, San Marino. "They all have very unique histories and the one thing that they are all very small" Full Article
ino AT#365 - Travel to the South Side of Chicago, Illinois By usa.amateurtraveler.com Published On :: Sat, 30 Mar 2013 17:58:33 +0000 Hear about travel to the South Side of Chicago as the Amateur Traveler talks to Amy Bugbee, author of "Suffering and Celebration of Life in America", about her hometown, the South Side of Chicago. The South Side of Chicago is more than just the White Sox and the stock yards just as Chicago pizza is more than just deep dish. Full Article
ino AT#482 - Hiking the Camino de Santiago in Spain By europe.amateurtraveler.com Published On :: Sat, 05 Sep 2015 15:14:47 +0000 Hear about hiking the Camino de Santiago (the Way of St. James) in northern Spain as the Amateur Traveler talks to Linda Martin from The Indie Travel Podcast. Linda says, "the thing about the Camino is that it is as long as you wanted to be." Full Article
ino AT#568 - Hiking the Camino Ronda in Northern Spain By europe.amateurtraveler.com Published On :: Sat, 22 Jul 2017 14:00:00 +0000 Hear about hiking the Camino Ronda in Northern Spain as the Amateur Traveler talks to Sherry Ott about her trek along the coastline of the Costa Brava region. Full Article
ino AT#597 - Travel to Chicago, Illinois By usa.amateurtraveler.com Published On :: Sat, 03 Mar 2018 15:00:00 +0000 Hear about travel to Chicago, Illinois as the Amateur Traveler talks to Francesca Mazurkiewicz about her hometown of Chicago. Full Article
ino Watch How One Freedom Caucus Member Sees the GOP’s Latino Voter Problem By feedproxy.google.com Published On :: Tue, 20 Oct 2015 11:06:32 EST "We're writing off too many people," Rep. Mick Mulvaney (R-SC) says in "Immigration Battle," a feature film presentation from FRONTLINE and Independent Lens that airs tonight on PBS. Full Article
ino How to Find Online Casino Bonuses Suited for You By feedproxy.google.com Published On :: Fri, 01 May 2020 13:55:28 +0000 Online casinos are famous for their bonuses and promotions. These offers are one of the key advantages that online casinos hold over their land-based equivalents. But there are a lot of different types of online casino bonuses. Anyone who is quite new to gambling online might be confused about how they work in practice. As […] The post How to Find Online Casino Bonuses Suited for You appeared first on Dumb Little Man. Full Article How To Money
ino 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
ino Yojiro Uchino By feedproxy.google.com Published On :: Fri, 02 Aug 2019 09:37:37 +0000 Visiting Fellow, Global Economy and Finance Programme Biography Yojiro Uchino was director of the defence budget at the Ministry of Finance in Japan from 2016 to 2019, working on budgets for the country's National Defense Program Guidelines and also its Mid-Term Defense Program.During his fellowship, Yojiro will be undertaking research on the relationship between national security and fiscal positions, as well as the balance between free trade and national security.Yojiro Uchino is based at Chatham House until July 2021, hosted by the Global Economy and Finance programme. Areas of expertise Budget structure of the Japanese governmentDefence budget of JapanJapanese defence policy Past experience 2015-16 Director, Allowance Control and Mutual Assistance Insurance Division2014-15 Director, Inter-Division Affairs of Budget2012-14 Director, Government Shareholding Office (where he planned simultaneous IPO of Japan Post, the holding company, and its subsidiaries Japan Post Bank and Japan Post Insurance)1997Admitted to the Bar in New York State Supreme Court1996LLM, University of Michigan Law School1992BA Law, University of Tokyo +44 (0)20 7314 2776 Email Full Article
ino 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
ino {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
ino Expanding Sino–Maghreb Relations: Morocco and Tunisia By feedproxy.google.com Published On :: Wed, 26 Feb 2020 12:49:01 +0000 26 February 2020 Over the past two decades, China has increased its presence in North Africa in terms of trade and investment. This paper looks at China’s policy within the context of its Africa and Middle East policies to better understand its approach to Morocco and Tunisia. Read online Download PDF Yahia H. Zoubir Senior Professor of International Studies, KEDGE Business School, France and Visiting Fellow, Brookings Doha Center 2020-02-26-Xi-Jinping-King-Mohammed.jpg Chinese President Xi Jinping and King Mohammed VI of Morocco wave during a welcoming ceremony outside the Great Hall of the People in Beijing on 11 May 2016. Photo: Getty Images. SummaryChina’s presence in the Maghreb has increased in recent years, raising concerns among Western powers. China has focused on bilateral relations with these countries while also working within the Forum on China–Africa Cooperation (FOCAC) and the China–Arab States Cooperation Forum (CASCF). However, this engagement has limited strategic value compared to relations China has with Saudi Arabia or the United Arab Emirates.Since the launch of the Belt and Road Initiative (BRI) in 2013, China has shown greater interest in the Maghreb as an entry point to European and African markets. China has pursued commercial relations over political influence in the region. Morocco and Tunisia are still dependent on France, their former colonial power, and the European Union, which exert great political, economic, security and cultural influence over the two countries.The Maghreb countries’ economic relations with China have grown exponentially, with Algeria forming the closest relationship. However, Morocco and Tunisia are keen to attract China’s investment and involvement in major construction and infrastructure projects to boost industrial and economic development. While China’s investments in Morocco and Tunisia remain at a low level, trade relations with both countries have grown steadily. Politically, China’s policy of noninterference in domestic affairs appeals to Maghreb states, which resent Western interference.China’s influence in the Maghreb remains minimal. Its soft power push has struggled to promote advantages of strong relations with China beyond economics. Furthermore, Morocco and Tunisia’s populations have generally scant knowledge about China’s politics and culture. China has tried to address this lack of familiarity through the establishment of Confucius Institutes and other cultural activities. However, language and cultural barriers still impede the development of close relations, compared to those China has with other countries in Africa. Department/project Middle East and North Africa Programme, Economic Transformation in North Africa Full Article
ino Extending the Limits of Quantitative Proteome Profiling with Data-Independent Acquisition and Application to Acetaminophen-Treated Three-Dimensional Liver Microtissues By feedproxy.google.com Published On :: 2015-05-01 Roland BrudererMay 1, 2015; 14:1400-1410Research Full Article
ino Discordant Protein and mRNA Expression in Lung Adenocarcinomas By feedproxy.google.com Published On :: 2002-04-01 Guoan ChenApr 1, 2002; 1:304-313Research Full Article
ino Stable Isotope Labeling by Amino Acids in Cell Culture, SILAC, as a Simple and Accurate Approach to Expression Proteomics By feedproxy.google.com Published On :: 2002-05-01 Shao-En OngMay 1, 2002; 1:376-386Research Full Article
ino The cytochrome P450 enzyme CYP24A1 increases proliferation of mutant KRAS-dependent lung adenocarcinoma independent of its catalytic activity [Cell Biology] By feedproxy.google.com Published On :: 2020-05-01T00:06:09-07:00 We previously reported that overexpression of cytochrome P450 family 24 subfamily A member 1 (CYP24A1) increases lung cancer cell proliferation by activating RAS signaling and that CYP24A1 knockdown inhibits tumor growth. However, the mechanism of CYP24A1-mediated cancer cell proliferation remains unclear. Here, we conducted cell synchronization and biochemical experiments in lung adenocarcinoma cells, revealing a link between CYP24A1 and anaphase-promoting complex (APC), a key cell cycle regulator. We demonstrate that CYP24A1 expression is cell cycle–dependent; it was higher in the G2-M phase and diminished upon G1 entry. CYP24A1 has a functional destruction box (D-box) motif that allows binding with two APC adaptors, CDC20-homologue 1 (CDH1) and cell division cycle 20 (CDC20). Unlike other APC substrates, however, CYP24A1 acted as a pseudo-substrate, inhibiting CDH1 activity and promoting mitotic progression. Conversely, overexpression of a CYP24A1 D-box mutant compromised CDH1 binding, allowing CDH1 hyperactivation, thereby hastening degradation of its substrates cyclin B1 and CDC20, and accumulation of the CDC20 substrate p21, prolonging mitotic exit. These activities also occurred with a CYP24A1 isoform 2 lacking the catalytic cysteine (Cys-462), suggesting that CYP24A1's oncogenic potential is independent of its catalytic activity. CYP24A1 degradation reduced clonogenic survival of mutant KRAS-driven lung cancer cells, and calcitriol treatment increased CYP24A1 levels and tumor burden in Lsl-KRASG12D mice. These results disclose a catalytic activity-independent growth-promoting role of CYP24A1 in mutant KRAS-driven lung cancer. This suggests that CYP24A1 could be therapeutically targeted in lung cancers in which its expression is high. Full Article
ino Biochemical and structural insights into how amino acids regulate pyruvate kinase muscle isoform 2 [Enzymology] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 Pyruvate kinase muscle isoform 2 (PKM2) is a key glycolytic enzyme involved in ATP generation and critical for cancer metabolism. PKM2 is expressed in many human cancers and is regulated by complex mechanisms that promote tumor growth and proliferation. Therefore, it is considered an attractive therapeutic target for modulating tumor metabolism. Various stimuli allosterically regulate PKM2 by cycling it between highly active and less active states. Several small molecules activate PKM2 by binding to its intersubunit interface. Serine and cysteine serve as an activator and inhibitor of PKM2, respectively, by binding to its amino acid (AA)-binding pocket, which therefore represents a potential druggable site. Despite binding similarly to PKM2, how cysteine and serine differentially regulate this enzyme remains elusive. Using kinetic analyses, fluorescence binding, X-ray crystallography, and gel filtration experiments with asparagine, aspartate, and valine as PKM2 ligands, we examined whether the differences in the side-chain polarity of these AAs trigger distinct allosteric responses in PKM2. We found that Asn (polar) and Asp (charged) activate PKM2 and that Val (hydrophobic) inhibits it. The results also indicate that both Asn and Asp can restore the activity of Val-inhibited PKM2. AA-bound crystal structures of PKM2 displayed distinctive interactions within the binding pocket, causing unique allosteric effects in the enzyme. These structure-function analyses of AA-mediated PKM2 regulation shed light on the chemical requirements in the development of mechanism-based small-molecule modulators targeting the AA-binding pocket of PKM2 and provide broader insights into the regulatory mechanisms of complex allosteric enzymes. Full Article
ino An enzyme-based protocol for cell-free synthesis of nature-identical capsular oligosaccharides from Actinobacillus pleuropneumoniae serotype 1 [Enzymology] By feedproxy.google.com Published On :: 2020-04-24T06:08:45-07:00 Actinobacillus pleuropneumoniae (App) is the etiological agent of acute porcine pneumonia and responsible for severe economic losses worldwide. The capsule polymer of App serotype 1 (App1) consists of [4)-GlcNAc-β(1,6)-Gal-α-1-(PO4-] repeating units that are O-acetylated at O-6 of the GlcNAc. It is a major virulence factor and was used in previous studies in the successful generation of an experimental glycoconjugate vaccine. However, the application of glycoconjugate vaccines in the animal health sector is limited, presumably because of the high costs associated with harvesting the polymer from pathogen culture. Consequently, here we exploited the capsule polymerase Cps1B of App1 as an in vitro synthesis tool and an alternative for capsule polymer provision. Cps1B consists of two catalytic domains, as well as a domain rich in tetratricopeptide repeats (TPRs). We compared the elongation mechanism of Cps1B with that of a ΔTPR truncation (Cps1B-ΔTPR). Interestingly, the product profiles displayed by Cps1B suggested processive elongation of the nascent polymer, whereas Cps1B-ΔTPR appeared to work in a more distributive manner. The dispersity of the synthesized products could be reduced by generating single-action transferases and immobilizing them on individual columns, separating the two catalytic activities. Furthermore, we identified the O-acetyltransferase Cps1D of App1 and used it to modify the polymers produced by Cps1B. Two-dimensional NMR analyses of the products revealed O-acetylation levels identical to those of polymer harvested from App1 culture supernatants. In conclusion, we have established a protocol for the pathogen-free in vitro synthesis of tailored, nature-identical App1 capsule polymers. Full Article
ino A Legionella effector kinase is activated by host inositol hexakisphosphate [Enzymology] By feedproxy.google.com Published On :: 2020-05-01T00:06:09-07:00 The transfer of a phosphate from ATP to a protein substrate, a modification known as protein phosphorylation, is catalyzed by protein kinases. Protein kinases play a crucial role in virtually every cellular activity. Recent studies of atypical protein kinases have highlighted the structural similarity of the kinase superfamily despite notable differences in primary amino acid sequence. Here, using a bioinformatics screen, we searched for putative protein kinases in the intracellular bacterial pathogen Legionella pneumophila and identified the type 4 secretion system effector Lpg2603 as a remote member of the protein kinase superfamily. Employing an array of biochemical and structural biology approaches, including in vitro kinase assays and isothermal titration calorimetry, we show that Lpg2603 is an active protein kinase with several atypical structural features. Importantly, we found that the eukaryote-specific host signaling molecule inositol hexakisphosphate (IP6) is required for Lpg2603 kinase activity. Crystal structures of Lpg2603 in the apo-form and when bound to IP6 revealed an active-site rearrangement that allows for ATP binding and catalysis. Our results on the structure and activity of Lpg2603 reveal a unique mode of regulation of a protein kinase, provide the first example of a bacterial kinase that requires IP6 for its activation, and may aid future work on the function of this effector during Legionella pathogenesis. Full Article
ino Roles of active-site residues in catalysis, substrate binding, cooperativity, and the reaction mechanism of the quinoprotein glycine oxidase [Enzymology] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 The quinoprotein glycine oxidase from the marine bacterium Pseudoalteromonas luteoviolacea (PlGoxA) uses a protein-derived cysteine tryptophylquinone (CTQ) cofactor to catalyze conversion of glycine to glyoxylate and ammonia. This homotetrameric enzyme exhibits strong cooperativity toward glycine binding. It is a good model for studying enzyme kinetics and cooperativity, specifically for being able to separate those aspects of protein function through directed mutagenesis. Variant proteins were generated with mutations in four active-site residues, Phe-316, His-583, Tyr-766, and His-767. Structures for glycine-soaked crystals were obtained for each. Different mutations had differential effects on kcat and K0.5 for catalysis, K0.5 for substrate binding, and the Hill coefficients describing the steady-state kinetics or substrate binding. Phe-316 and Tyr-766 variants retained catalytic activity, albeit with altered kinetics and cooperativity. Substitutions of His-583 revealed that it is essential for glycine binding, and the structure of H583C PlGoxA had no active-site glycine present in glycine-soaked crystals. The structure of H767A PlGoxA revealed a previously undetected reaction intermediate, a carbinolamine product-reduced CTQ adduct, and exhibited only negligible activity. The results of these experiments, as well as those with the native enzyme and previous variants, enabled construction of a detailed mechanism for the reductive half-reaction of glycine oxidation. This proposed mechanism includes three discrete reaction intermediates that are covalently bound to CTQ during the reaction, two of which have now been structurally characterized by X-ray crystallography. Full Article
ino A single amino acid substitution uncouples catalysis and allostery in an essential biosynthetic enzyme in Mycobacterium tuberculosis [Enzymology] By feedproxy.google.com Published On :: 2020-05-08T03:41:14-07:00 Allostery exploits the conformational dynamics of enzymes by triggering a shift in population ensembles toward functionally distinct conformational or dynamic states. Allostery extensively regulates the activities of key enzymes within biosynthetic pathways to meet metabolic demand for their end products. Here, we have examined a critical enzyme, 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAH7PS), at the gateway to aromatic amino acid biosynthesis in Mycobacterium tuberculosis, which shows extremely complex dynamic allostery: three distinct aromatic amino acids jointly communicate occupancy to the active site via subtle changes in dynamics, enabling exquisite fine-tuning of delivery of these essential metabolites. Furthermore, this allosteric mechanism is co-opted by pathway branchpoint enzyme chorismate mutase upon complex formation. In this study, using statistical coupling analysis, site-directed mutagenesis, isothermal calorimetry, small-angle X-ray scattering, and X-ray crystallography analyses, we have pinpointed a critical node within the complex dynamic communication network responsible for this sophisticated allosteric machinery. Through a facile Gly to Pro substitution, we have altered backbone dynamics, completely severing the allosteric signal yet remarkably, generating a nonallosteric enzyme that retains full catalytic activity. We also identified a second residue of prime importance to the inter-enzyme communication with chorismate mutase. Our results reveal that highly complex dynamic allostery is surprisingly vulnerable and provide further insights into the intimate link between catalysis and allostery. Full Article
ino Nowhere to Call Home: Ethnic Minorities in China By feedproxy.google.com Published On :: Mon, 26 Jan 2015 11:15:01 +0000 Members Event 18 February 2015 - 6:00pm to 7:15pm Chatham House, London Transcriptpdf | 147.75 KB Transcript Q&Apdf | 160.08 KB Event participants Jocelyn Ford, Journalist and Filmmaker, Nowhere to Call Home: A Tibetan in BeijingDr Reza Hasmath, Lecturer in Chinese Politics, University of OxfordChair: Rob Gifford, Correspondent, The Economist Jocelyn Ford will share her experiences and insights from documenting the struggles of a widowed Tibetan facing ethnic discrimination in Beijing and gender discrimination in her village. The panel will then have a wider discussion about national identity and the issues facing ethnic minorities in China. This discussion coincides with the UK screening of Nowhere to Call Home: A Tibetan in Beijing and will include clips from the film. Full Article
ino Gene expression regulation by retinoic acid By feedproxy.google.com Published On :: 2002-11-01 James E. BalmerNov 1, 2002; 43:1773-1808Reviews Full Article
ino {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
ino Identification of an Unconventional Subpeptidome Bound to the Behcet's Disease-associated HLA-B*51:01 that is Regulated by Endoplasmic Reticulum Aminopeptidase 1 (ERAP1) [Research] By feedproxy.google.com Published On :: 2020-05-01T00:05:26-07:00 Human leukocyte antigen (HLA) B*51:01 and endoplasmic reticulum aminopeptidase 1 (ERAP1) are strongly genetically associated with Behcet's disease (BD). Previous studies have defined two subgroups of HLA-B*51 peptidome containing proline (Pro) or alanine (Ala) at position 2 (P2). Little is known about the unconventional non-Pro/Ala2 HLA-B*51-bound peptides. We aimed to study the features of this novel subpeptidome, and investigate its regulation by ERAP1. CRISPR-Cas9 was used to generate an HLA-ABC-triple knockout HeLa cell line (HeLa.ABC-KO), which was subsequently transduced to express HLA-B*51:01 (HeLa.ABC-KO.B51). ERAP1 was silenced using lentiviral shRNA. Peptides bound to HLA-B*51:01 were eluted and analyzed by mass spectrometry. The characteristics of non-Pro/Ala2, Pro2, and Ala2 peptides and their alteration by ERAP1 silencing were investigated. Effects of ERAP1 silencing on cell surface expression of HLA-B*51:01 were studied using flow cytometry. More than 20% of peptides eluted from HLA-B*51:01 lacked Pro or Ala at P2. This unconventional group of HLA-B*51:01-bound peptides was relatively enriched for 8-mers (with relatively fewer 9-mers) compared with the Pro2 and Ala2 subpeptidomes and had similar N-terminal and C-terminal residue usages to Ala2 peptides (with the exception of the less abundant leucine at position ). Knockdown of ERAP1 increased the percentage of non-Pro/Ala2 from 20% to ~40%, increased the percentage of longer (10-mer and 11-mer) peptides eluted from HLA-B*51:01 complexes, and abrogated the predominance of leucine at P1. Interestingly knockdown of ERAP1 altered the length and N-terminal residue usage of non-Ala2&Pro2 and Ala2 but not the Pro2 peptides. Finally, ERAP1 silencing regulated the expression levels of cell surface HLA-B*51 in a cell-type-dependent manner. In conclusion, we have used a novel methodology to identify an unconventional but surprisingly abundant non-Pro/Ala2 HLA-B*51:01 subpeptidome. It is increased by knockdown of ERAP1, a gene affecting the risk of developing BD. This has implications for theories of disease pathogenesis. Full Article
ino Biochemical and structural insights into how amino acids regulate pyruvate kinase muscle isoform 2 [Enzymology] By feedproxy.google.com Published On :: 2020-04-17T00:06:05-07:00 Pyruvate kinase muscle isoform 2 (PKM2) is a key glycolytic enzyme involved in ATP generation and critical for cancer metabolism. PKM2 is expressed in many human cancers and is regulated by complex mechanisms that promote tumor growth and proliferation. Therefore, it is considered an attractive therapeutic target for modulating tumor metabolism. Various stimuli allosterically regulate PKM2 by cycling it between highly active and less active states. Several small molecules activate PKM2 by binding to its intersubunit interface. Serine and cysteine serve as an activator and inhibitor of PKM2, respectively, by binding to its amino acid (AA)-binding pocket, which therefore represents a potential druggable site. Despite binding similarly to PKM2, how cysteine and serine differentially regulate this enzyme remains elusive. Using kinetic analyses, fluorescence binding, X-ray crystallography, and gel filtration experiments with asparagine, aspartate, and valine as PKM2 ligands, we examined whether the differences in the side-chain polarity of these AAs trigger distinct allosteric responses in PKM2. We found that Asn (polar) and Asp (charged) activate PKM2 and that Val (hydrophobic) inhibits it. The results also indicate that both Asn and Asp can restore the activity of Val-inhibited PKM2. AA-bound crystal structures of PKM2 displayed distinctive interactions within the binding pocket, causing unique allosteric effects in the enzyme. These structure-function analyses of AA-mediated PKM2 regulation shed light on the chemical requirements in the development of mechanism-based small-molecule modulators targeting the AA-binding pocket of PKM2 and provide broader insights into the regulatory mechanisms of complex allosteric enzymes. Full Article
ino Who can give me a theme similar to iNove ? By forums.digitalpoint.com Published On :: Fri, 08 May 2020 18:26:23 +0000 Full Article
ino University spinouts: processes, benefits and risks of the system By www.techworld.com Published On :: Tue, 03 Sep 2019 13:08:00 GMT Spinout founders, investors and insiders give their tips on turning academic research into successful businesses Full Article
ino How Queen's Belfast became the UK's best spinout university By www.techworld.com Published On :: Mon, 11 Nov 2019 15:16:00 GMT Researchers from the venture capital firm Octopus Ventures gave much of the credit to QUBIS, the university's dedicated commercialisation arm Full Article
ino CBD News: Mensaje de Sr. Braulio Ferreira de Souza Dias, Secretario Ejecutivo del CDB, en la ocasión de la XXIX asamblea ordinaria parlamento latinoamericano, Panamá, 18 -19 Octubre de 2013 By www.cbd.int Published On :: Fri, 18 Oct 2013 00:00:00 GMT Full Article
ino 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 www.jbc.org 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
ino Biochemical and structural insights into how amino acids regulate pyruvate kinase muscle isoform 2 [Enzymology] By www.jbc.org Published On :: 2020-04-17T00:06:05-07:00 Pyruvate kinase muscle isoform 2 (PKM2) is a key glycolytic enzyme involved in ATP generation and critical for cancer metabolism. PKM2 is expressed in many human cancers and is regulated by complex mechanisms that promote tumor growth and proliferation. Therefore, it is considered an attractive therapeutic target for modulating tumor metabolism. Various stimuli allosterically regulate PKM2 by cycling it between highly active and less active states. Several small molecules activate PKM2 by binding to its intersubunit interface. Serine and cysteine serve as an activator and inhibitor of PKM2, respectively, by binding to its amino acid (AA)-binding pocket, which therefore represents a potential druggable site. Despite binding similarly to PKM2, how cysteine and serine differentially regulate this enzyme remains elusive. Using kinetic analyses, fluorescence binding, X-ray crystallography, and gel filtration experiments with asparagine, aspartate, and valine as PKM2 ligands, we examined whether the differences in the side-chain polarity of these AAs trigger distinct allosteric responses in PKM2. We found that Asn (polar) and Asp (charged) activate PKM2 and that Val (hydrophobic) inhibits it. The results also indicate that both Asn and Asp can restore the activity of Val-inhibited PKM2. AA-bound crystal structures of PKM2 displayed distinctive interactions within the binding pocket, causing unique allosteric effects in the enzyme. These structure-function analyses of AA-mediated PKM2 regulation shed light on the chemical requirements in the development of mechanism-based small-molecule modulators targeting the AA-binding pocket of PKM2 and provide broader insights into the regulatory mechanisms of complex allosteric enzymes. Full Article
ino Proline-rich 11 (PRR11) drives F-actin assembly by recruiting the actin-related protein 2/3 complex in human non-small cell lung carcinoma [DNA and Chromosomes] By www.jbc.org Published On :: 2020-04-17T00:06:05-07:00 The actin cytoskeleton is extremely dynamic and supports diverse cellular functions in many physiological and pathological processes, including tumorigenesis. However, the mechanisms that regulate the actin-related protein 2/3 (ARP2/3) complex and thereby promote actin polymerization and organization in cancer cells are not well-understood. We previously implicated the proline-rich 11 (PRR11) protein in lung cancer development. In this study, using immunofluorescence staining, actin polymerization assays, and siRNA-mediated gene silencing, we uncovered that cytoplasmic PRR11 is involved in F-actin polymerization and organization. We found that dysregulation of PRR11 expression results in F-actin rearrangement and nuclear instability in non-small cell lung cancer cells. Results from molecular mechanistic experiments indicated that PRR11 associates with and recruits the ARP2/3 complex, facilitates F-actin polymerization, and thereby disrupts the F-actin cytoskeleton, leading to abnormal nuclear lamina assembly and chromatin reorganization. Inhibition of the ARP2/3 complex activity abolished irregular F-actin polymerization, lamina assembly, and chromatin reorganization due to PRR11 overexpression. Notably, experiments with truncated PRR11 variants revealed that PRR11 regulates F-actin through different regions. We found that deletion of either the N or C terminus of PRR11 abrogates its effects on F-actin polymerization and nuclear instability and that deletion of amino acid residues 100–184 or 100–200 strongly induces an F-actin structure called the actin comet tail, not observed with WT PRR11. Our findings indicate that cytoplasmic PRR11 plays an essential role in regulating F-actin assembly and nuclear stability by recruiting the ARP2/3 complex in human non-small cell lung carcinoma cells. Full Article
ino An enzyme-based protocol for cell-free synthesis of nature-identical capsular oligosaccharides from Actinobacillus pleuropneumoniae serotype 1 [Enzymology] By www.jbc.org Published On :: 2020-04-24T06:08:45-07:00 Actinobacillus pleuropneumoniae (App) is the etiological agent of acute porcine pneumonia and responsible for severe economic losses worldwide. The capsule polymer of App serotype 1 (App1) consists of [4)-GlcNAc-β(1,6)-Gal-α-1-(PO4-] repeating units that are O-acetylated at O-6 of the GlcNAc. It is a major virulence factor and was used in previous studies in the successful generation of an experimental glycoconjugate vaccine. However, the application of glycoconjugate vaccines in the animal health sector is limited, presumably because of the high costs associated with harvesting the polymer from pathogen culture. Consequently, here we exploited the capsule polymerase Cps1B of App1 as an in vitro synthesis tool and an alternative for capsule polymer provision. Cps1B consists of two catalytic domains, as well as a domain rich in tetratricopeptide repeats (TPRs). We compared the elongation mechanism of Cps1B with that of a ΔTPR truncation (Cps1B-ΔTPR). Interestingly, the product profiles displayed by Cps1B suggested processive elongation of the nascent polymer, whereas Cps1B-ΔTPR appeared to work in a more distributive manner. The dispersity of the synthesized products could be reduced by generating single-action transferases and immobilizing them on individual columns, separating the two catalytic activities. Furthermore, we identified the O-acetyltransferase Cps1D of App1 and used it to modify the polymers produced by Cps1B. Two-dimensional NMR analyses of the products revealed O-acetylation levels identical to those of polymer harvested from App1 culture supernatants. In conclusion, we have established a protocol for the pathogen-free in vitro synthesis of tailored, nature-identical App1 capsule polymers. Full Article
ino A Legionella effector kinase is activated by host inositol hexakisphosphate [Enzymology] By www.jbc.org Published On :: 2020-05-01T00:06:09-07:00 The transfer of a phosphate from ATP to a protein substrate, a modification known as protein phosphorylation, is catalyzed by protein kinases. Protein kinases play a crucial role in virtually every cellular activity. Recent studies of atypical protein kinases have highlighted the structural similarity of the kinase superfamily despite notable differences in primary amino acid sequence. Here, using a bioinformatics screen, we searched for putative protein kinases in the intracellular bacterial pathogen Legionella pneumophila and identified the type 4 secretion system effector Lpg2603 as a remote member of the protein kinase superfamily. Employing an array of biochemical and structural biology approaches, including in vitro kinase assays and isothermal titration calorimetry, we show that Lpg2603 is an active protein kinase with several atypical structural features. Importantly, we found that the eukaryote-specific host signaling molecule inositol hexakisphosphate (IP6) is required for Lpg2603 kinase activity. Crystal structures of Lpg2603 in the apo-form and when bound to IP6 revealed an active-site rearrangement that allows for ATP binding and catalysis. Our results on the structure and activity of Lpg2603 reveal a unique mode of regulation of a protein kinase, provide the first example of a bacterial kinase that requires IP6 for its activation, and may aid future work on the function of this effector during Legionella pathogenesis. Full Article
