ino Dage Minors Wins Three Brooks Parkrun In UK By bernews.com Published On :: Tue, 17 Sep 2024 14:59:27 +0000 Dage Minors ran to victory in the 2024 Three Brooks Parkrun in the UK. Minors crossed the line in a time of 17:10 to defeat a field of 345 competitors. Related Stories Watson & Beaulieu Win Swan’s Charity 5K Ryan Outerbridge Wins 8k In Massachusetts Richardson Wins Long Beach 5 Mile Road Race Ryan Outerbridge […] Full Article All Sports #BermudaRoadRunning #GoodNews
ino Review: A Night Of Jazz With Tino Martinez By bernews.com Published On :: Thu, 03 Oct 2024 14:06:42 +0000 [Written by Dale Butler] Tino Martinez is now broadening his Bermuda music experience with a variety of guest artists. On Thursday, September 26 at 7:30pm at The Deep inside the Lido Complex [Elbow Beach], Tino pulled out all stops. Regulars in the band, Raymond George [piano] and Torey Tacklin [bass] had a joyous evening with […] Full Article All Entertainment Music #DaleButler #DaleButlerColumns #Music
ino Did Oil Kill the Dinosaurs? By Published On :: Thu, 28 Jul 2016 10:51:00 GMT Sixty-six million years ago, planet Earth was forever changed when a six-mile-wide asteroid smashed into the Yucatan Peninsula, triggering a series of events that killed off the dinosaurs. Full Article
ino Your impact: September equinox 2024 By www.planetary.org Published On :: Mon, 09 Sep 2024 06:56:00 -0700 Exploring Europa and defending Earth. Full Article
ino Older people from black and minority ethnic backgrounds Accessing health and social care services in south GLasgow By lx.iriss.org.uk Published On :: Wednesday, August 12, 2015 - 11:54 The Advocacy Project works with older people and other groups across Glasgow and Lanarkshire to ensure their voice is heard, their needs met and their legal rights safeguarded. The organisation identified a low take up of their own service by older people from BME communities, which was generally held to reflect the wider picture in Glasgow in relation to health and social work services. This report was commissioned by them to examine barriers to access to services specifically for older people from BME communities in Glasgow South, where there is a high concentration of people from BME communities. Full Article
ino FAFO: Latino Believes Trump Won't Deport Family-Oriented Migrants By crooksandliars.com Published On :: Wed, 13 Nov 2024 02:59:24 +0000 In today's edition of FAFO (F*ck Around and Find Out) we have a man who is probably going to learn the hard way that being one of the "good ones" means nothing to the Trump administration. See, MAGA loyalty only goes one way. They want your vote, but won't do anything to make your life better. This Latino male Trump voter told CNN that mass deportation won't extend to law-abiding workers. He actually said, with a straight face: “That wouldn’t be fair. They need to make sure that they don’t throw away, they don’t kick out, they don’t deport people that are family oriented.” Sweetie, they don't care if you are family oriented or not. If you are working a good job or not. If you are a criminal or not. To them, being non-American is enough to get you kicked out (and probably put in a camp before). Enjoy that Trump hair visor hat, honey. The leopards are coming for your face. "Fucketh around and ye shall findeth out" - A Twitter User in 2024 read more Full Article deportation FAFO Latino Leopards Eating Faces MAGA
ino Apparently, Basic Terminology Isn't Basic For Everyone By clientsfromhell.tumblr.com Published On :: Sun, 20 Mar 2022 08:00:31 -0700 Client: “I need the number for Jack. Can you give me his number, please?” Me: “Jack? Jack who?” Client: “The manual for the modem said that I need to connect to Jack, so if you could give me his number, then I can connect with him.” Full Article
ino Winos 4.0 Malware Infects Gamers Through Malicious Game Optimization Apps By thehackernews.com Published On :: Wed, 06 Nov 2024 19:29:00 +0530 Cybersecurity researchers are warning that a command-and-control (C&C) framework called Winos is being distributed within gaming-related applications like installation tools, speed boosters, and optimization utilities. "Winos 4.0 is an advanced malicious framework that offers comprehensive functionality, a stable architecture, and efficient control over numerous online endpoints to execute Full Article
ino Our lastest digital signage case study comes from Illinois in the US. By www.repeatsoftware.com Published On :: Mon, 30 Nov 2020 00:59:46 GMT Minerallac Company, an electrical construction hardware manufacturer, find Repeat Signage software a useful tool for keeping staff updated on events and information. Staff is their satellite office, 2000 miles away, now feel more connected since digital signage was utilised. Full Article
ino O que Israel, Netanyahu e os palestinos devem esperar do governo Trump By www.bbc.com Published On :: Sun, 10 Nov 2024 19:11:48 GMT A vitória de Donald Trump foi bem recebida em Israel, mas a mudança que ele traz pode não ser do agrado do país. Full Article
ino Liberal Media: “Minority Racists” Elected Trump By theblacksphere.net Published On :: Tue, 12 Nov 2024 19:50:22 +0000 The following article, Liberal Media: “Minority Racists” Elected Trump, was first published on The Black Sphere. A Message Sent No One Heard Donald Trump is the President-elect for the second time! His will be the second split-presidency incumbent nominee since Grover Cleveland. But last Tuesday night was not just 312-to-226 Electoral College blowout, but with Trump receiving 74,708,855 (and growing) vs Vice President Kamala Harris’ 70,980,347. The former 45th President also … Continue reading Liberal Media: “Minority Racists” Elected Trump ... Full Article Democrats Election of 2024 Featured #DonaldTrump #Trump2024 #Womenvote 2024 election black vote black voters black vs white electoral college Hispanic vote Joe Scarborough Kamala Harris the black vote The View War on Women
ino Say it ain’t so, Joe! Latino voters on Telemundo give President Trump the debate win 66% to 34% By powderedwigsociety.com Published On :: Thu, 01 Oct 2020 00:37:49 +0000 Suddenly, the Democratic Party is all about stopping illegal immigration. The post Say it ain’t so, Joe! Latino voters on Telemundo give President Trump the debate win 66% to 34% appeared first on Powdered Wig Society. Full Article 2020 presidential election telemundo
ino Teacher at Chino Valley School District Goes Off the Rails over a Student With a Trump Hat By lidblog.com Published On :: Tue, 12 Nov 2024 13:56:14 +0000 A “teacher” at Chino Valley School District launched into an unhinged rant after a student entered his classroom wearing a Trump hat. His name is Clyde J Colinco, who is also the girls golf course, and unfortunately, he still appears to be employed at Chino High School. “Trump Derangement Syndrome is a real thing. If […] The post Teacher at Chino Valley School District Goes Off the Rails over a Student With a Trump Hat appeared first on The Lid. Full Article Woke
ino Paleontologist rates 14 dinosaur attacks in the "Jurassic Park" movies By www.businessinsider.com Published On :: Tue, 12 Nov 2024 19:00:02 +0000 Paleontologist Dave Hone rates the realism of the dinosaur attacks in the "Jurassic Park" and "Jurassic World" movies. Full Article Entertainment video video-format-how-real-is-it how-real-is-it movies dinosaurs paleontology jurassic-park jurassic-world jurassic-world-2 jurassic-world-3 jurassic-world-dominion jurassic-world-fallen-kingdom jurassic-park-2 jurassic-park-3 steven-spielberg laura-dern sam-neill jeff-goldblum chris-pratt bryce-dallas-howard ja-bayona science julianne-moore vince-vaughn william-h-macy new-this-week-video
ino Top 3 Airports With Casinos By www.upgradetravelbetter.com Published On :: Sat, 27 May 2017 08:03:32 +0000 Have you ever been at an airport waiting to connect your flight for hours on end? We all have, and yeah, it always blows to twiddle your thumbs for hours waiting for a flight that might delay or not. Aside from drinking mugs after mugs of coffee, there really doesn’t seem to be much one […] The post Top 3 Airports With Casinos first appeared on UPGRADE: TRAVEL BETTER. Full Article Travel
ino The Best Casinos to Visit When You Travel To New Zealand By www.upgradetravelbetter.com Published On :: Thu, 05 Apr 2018 16:00:20 +0000 Thanks to a change in the laws regulating gambling, land casinos are becoming popular in New Zealand. Today, the country has six casinos in total. Due to the relatively small number of casinos in the country, the industry focuses on offering superior quality as opposed to quantity. Of course, online gambling has grown too, providing […] The post The Best Casinos to Visit When You Travel To New Zealand first appeared on UPGRADE: TRAVEL BETTER. Full Article Uncategorized
ino Top 3 Casinos You Should Visit When Flying to India By www.upgradetravelbetter.com Published On :: Tue, 28 Feb 2023 07:14:00 +0000 As a tourist destination, India has a lot to offer. From stunning nature, over unique culture, to interesting architecture, the country is a dream for any free-spirited traveler out there. Whatever it is that you’re after, from bustling cities to relaxing natural landscapes, you’ll find it here. But what about those who also enjoy trying […] The post Top 3 Casinos You Should Visit When Flying to India first appeared on UPGRADE: TRAVEL BETTER. Full Article Uncategorized
ino The Best Online Casinos to Gamble at While Flying By Plane By www.upgradetravelbetter.com Published On :: Sun, 09 Apr 2023 09:10:30 +0000 Are you looking for the perfect way to gamble while flying on a plane? Look no further – here are our picks for the top online casinos! Introduction Flying can be a long and exhausting process. Many travelers seek solace in online gambling, offering the convenience of playing casino games while on-the-go. Thanks to Wi-Fi […] The post The Best Online Casinos to Gamble at While Flying By Plane first appeared on UPGRADE: TRAVEL BETTER. Full Article Travel Travvies
ino Teacher at Chino Valley School District Goes Off the Rails over a Student With a Trump Hat By conservativefiringline.com Published On :: Mon, 11 Nov 2024 19:14:43 +0000 The following article, Teacher at Chino Valley School District Goes Off the Rails over a Student With a Trump Hat, was first published on Conservative Firing Line. A “teacher” at Chino Valley School District launched into an unhinged rant after a student entered his classroom wearing a Trump hat. His name is Clyde J Colinco, who is also the girls golf course, and unfortunately, he still appears to be employed at Chino High School. “Trump Derangement Syndrome is a real thing. If … Continue reading Teacher at Chino Valley School District Goes Off the Rails over a Student With a Trump Hat ... Full Article Opinion Politics C J Colinco Chino Valley golf coach teacher rant Trump hat
ino My Faith in Nerds: Stronger Than Any Gelatinous Cube By www.43folders.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
ino Disease-associated mutations in inositol 1,4,5-trisphosphate receptor subunits impair channel function [Molecular Bases of Disease] By www.jbc.org Published On :: 2020-12-25T00:06:30-08:00 The inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs), which form tetrameric channels, play pivotal roles in regulating the spatiotemporal patterns of intracellular calcium signals. Mutations in IP3Rs have been increasingly associated with many debilitating human diseases such as ataxia, Gillespie syndrome, and generalized anhidrosis. However, how these mutations affect IP3R function, and how the perturbation of as-sociated calcium signals contribute to the pathogenesis and severity of these diseases remains largely uncharacterized. Moreover, many of these diseases occur as the result of autosomal dominant inheritance, suggesting that WT and mutant subunits associate in heterotetrameric channels. How the in-corporation of different numbers of mutant subunits within the tetrameric channels affects its activities and results in different disease phenotypes is also unclear. In this report, we investigated representative disease-associated missense mutations to determine their effects on IP3R channel activity. Additionally, we designed concatenated IP3R constructs to create tetrameric channels with a predefined subunit composition to explore the functionality of heteromeric channels. Using calcium imaging techniques to assess IP3R channel function, we observed that all the mutations studied resulted in severely attenuated Ca2+ release when expressed as homotetramers. However, some heterotetramers retained varied degrees of function dependent on the composition of the tetramer. Our findings suggest that the effect of mutations depends on the location of the mutation in the IP3R structure, as well as on the stoichiometry of mutant subunits assembled within the tetrameric channel. These studies provide insight into the pathogenesis and penetrance of these devastating human diseases. Full Article
ino Extending the Limits of Quantitative Proteome Profiling with Data-Independent Acquisition and Application to Acetaminophen-Treated Three-Dimensional Liver Microtissues By www.mcponline.org Published On :: 2015-05-01 Roland BrudererMay 1, 2015; 14:1400-1410Research Full Article
ino Discordant Protein and mRNA Expression in Lung Adenocarcinomas By www.mcponline.org 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 www.mcponline.org Published On :: 2002-05-01 Shao-En OngMay 1, 2002; 1:376-386Research Full Article
ino Biosynthesis of the sactipeptide Ruminococcin C by the human microbiome: Mechanistic insights into thioether bond formation by radical SAM enzymes [Microbiology] By www.jbc.org Published On :: 2020-12-04T00:06:05-08:00 Despite its major importance in human health, the metabolic potential of the human gut microbiota is still poorly understood. We have recently shown that biosynthesis of Ruminococcin C (RumC), a novel ribosomally synthesized and posttranslationally modified peptide (RiPP) produced by the commensal bacterium Ruminococcus gnavus, requires two radical SAM enzymes (RumMC1 and RumMC2) catalyzing the formation of four Cα-thioether bridges. These bridges, which are essential for RumC's antibiotic properties against human pathogens such as Clostridium perfringens, define two hairpin domains giving this sactipeptide (sulfur-to-α-carbon thioether–containing peptide) an unusual architecture among natural products. We report here the biochemical and spectroscopic characterizations of RumMC2. EPR spectroscopy and mutagenesis data support that RumMC2 is a member of the large family of SPASM domain radical SAM enzymes characterized by the presence of three [4Fe-4S] clusters. We also demonstrate that this enzyme initiates its reaction by Cα H-atom abstraction and is able to catalyze the formation of nonnatural thioether bonds in engineered peptide substrates. Unexpectedly, our data support the formation of a ketoimine rather than an α,β-dehydro-amino acid intermediate during Cα-thioether bridge LC–MS/MS fragmentation. Finally, we explored the roles of the leader peptide and of the RiPP precursor peptide recognition element, present in myriad RiPP-modifying enzymes. Collectively, our data support a more complex role for the peptide recognition element and the core peptide for the installation of posttranslational modifications in RiPPs than previously anticipated and suggest a possible reaction intermediate for thioether bond formation. Full Article
ino Gene expression regulation by retinoic acid By www.jlr.org Published On :: 2002-11-01 James E. BalmerNov 1, 2002; 43:1773-1808Reviews Full Article
ino Sino-Russian Gas Cooperation: Power of Siberia I and II and Implications for Global LNG Supplies By www.chathamhouse.org Published On :: Tue, 19 Nov 2019 10:25:01 +0000 Sino-Russian Gas Cooperation: Power of Siberia I and II and Implications for Global LNG Supplies 27 November 2019 — 8:30AM TO 9:30AM Anonymous (not verified) 19 November 2019 Chatham House | 10 St James's Square | London | SW1Y 4LE In a new event in the Sustainable Transitions series, the speaker will present an update of Sino-Russian gas cooperation.To give a comprehensive account of their impact on global liquefied natural gas (LNG) supplies, he will discuss the following points:Gas is scheduled to start flowing from the Power of Siberia I (POS) on 2 December 2019. But what is the background of development of POS 1 and what is its current status and prospects? What are the chances of exporting gas through the proposed Altai pipeline? Why is the Mongolia export route so significant? And how will it affect the Central Asian Republics and in particular Turkmenistan’s gas export to China? What are the implications of both POS I and Altai gas via Mongolia route in the context of global LNG supply?What are the prospects for multilateral pipeline gas cooperation in northeast Asia?What are the implications for other Arctic onshore LNG supply, in particular, for Novatek’s Yamal LNG and Arctic LNG 1 and 2 to China on top of POS 1 and Altai gas?Attendance at this event is by invitation only. Full Article
ino Bessel periods and anticyclotomic ????-adic spinor ????-functions By www.ams.org Published On :: Thu, 31 Oct 2024 16:22 EDT Ming-Lun Hsieh and Shunsuke Yamana Trans. Amer. Math. Soc. 377 (), 5617-5672. Abstract, references and article information Full Article
ino Unique active-site and subsite features in the arabinogalactan-degrading GH43 exo-{beta}-1,3-galactanase from Phanerochaete chrysosporium [Enzymology] By www.jbc.org Published On :: 2020-12-25T00:06:31-08:00 Arabinogalactan proteins (AGPs) are plant proteoglycans with functions in growth and development. However, these functions are largely unexplored, mainly because of the complexity of the sugar moieties. These carbohydrate sequences are generally analyzed with the aid of glycoside hydrolases. The exo-β-1,3-galactanase is a glycoside hydrolase from the basidiomycete Phanerochaete chrysosporium (Pc1,3Gal43A), which specifically cleaves AGPs. However, its structure is not known in relation to its mechanism bypassing side chains. In this study, we solved the apo and liganded structures of Pc1,3Gal43A, which reveal a glycoside hydrolase family 43 subfamily 24 (GH43_sub24) catalytic domain together with a carbohydrate-binding module family 35 (CBM35) binding domain. GH43_sub24 is known to lack the catalytic base Asp conserved among other GH43 subfamilies. Our structure in combination with kinetic analyses reveals that the tautomerized imidic acid group of Gln263 serves as the catalytic base residue instead. Pc1,3Gal43A has three subsites that continue from the bottom of the catalytic pocket to the solvent. Subsite −1 contains a space that can accommodate the C-6 methylol of Gal, enabling the enzyme to bypass the β-1,6–linked galactan side chains of AGPs. Furthermore, the galactan-binding domain in CBM35 has a different ligand interaction mechanism from other sugar-binding CBM35s, including those that bind galactomannan. Specifically, we noted a Gly → Trp substitution, which affects pyranose stacking, and an Asp → Asn substitution in the binding pocket, which recognizes β-linked rather than α-linked Gal residues. These findings should facilitate further structural analysis of AGPs and may also be helpful in engineering designer enzymes for efficient biomass utilization. Full Article
ino MMP activation-associated aminopeptidase N reveals a bivalent 14-3-3 binding motif [Protein Structure and Folding] By www.jbc.org Published On :: 2020-12-25T00:06:31-08:00 Aminopeptidase N (APN, CD13) is a transmembrane ectopeptidase involved in many crucial cellular functions. Besides its role as a peptidase, APN also mediates signal transduction and is involved in the activation of matrix metalloproteinases (MMPs). MMPs function in tissue remodeling within the extracellular space and are therefore involved in many human diseases, such as fibrosis, rheumatoid arthritis, tumor angiogenesis, and metastasis, as well as viral infections. However, the exact mechanism that leads to APN-driven MMP activation is unclear. It was previously shown that extracellular 14-3-3 adapter proteins bind to APN and thereby induce the transcription of MMPs. As a first step, we sought to identify potential 14-3-3–binding sites in the APN sequence. We constructed a set of phosphorylated peptides derived from APN to probe for interactions. We identified and characterized a canonical 14-3-3–binding site (site 1) within the flexible, structurally unresolved N-terminal APN region using direct binding fluorescence polarization assays and thermodynamic analysis. In addition, we identified a secondary, noncanonical binding site (site 2), which enhances the binding affinity in combination with site 1 by many orders of magnitude. Finally, we solved crystal structures of 14-3-3σ bound to mono- and bis-phosphorylated APN-derived peptides, which revealed atomic details of the binding mode of mono- and bivalent 14-3-3 interactions. Therefore, our findings shed some light on the first steps of APN-mediated MMP activation and open the field for further investigation of this important signaling pathway. Full Article
ino Improved Localization of Insulinomas Using 68Ga-NODAGA-Exendin-4 PET/CT By jnm.snmjournals.org Published On :: 2024-10-17T09:03:09-07:00 Visual Abstract Full Article
ino CD70-Targeted Immuno-PET/CT Imaging of Clear Cell Renal Cell Carcinoma: A Translational Study By jnm.snmjournals.org Published On :: 2024-11-07T04:28:31-08:00 Visual Abstract Full Article
ino Kinome Profiling of Primary Endometrial Tumors Using Multiplexed Inhibitor Beads and Mass Spectrometry Identifies SRPK1 as Candidate Therapeutic Target By www.mcponline.org Published On :: 2020-12-01 Alison M. KurimchakDec 1, 2020; 19:2068-2089Research Full Article
ino Proteome analysis reveals a significant host-specific response in Rhizobium leguminosarum bv viciae endosymbiotic cells By www.mcponline.org Published On :: 2020-11-19 David DuránNov 19, 2020; 0:RA120.002276v1-mcp.RA120.002276Research Full Article
ino Proteomic analyses identify differentially expressed proteins and pathways between low-risk and high-risk subtypes of early-stage lung adenocarcinoma and their prognostic impacts By www.mcponline.org Published On :: 2020-11-30 Juntuo ZhouNov 30, 2020; 0:RA120.002384v1-mcp.RA120.002384Research Full Article
ino Lipid metabolism dysregulation in diabetic retinopathy By www.jlr.org Published On :: 2020-12-23 Julia V BusikDec 23, 2020; 0:jlr.TR120000981v1-jlr.TR120000981Thematic Reviews Full Article
ino Bisretinoid phospholipid and vitamin A aldehyde: Shining a light [Thematic Reviews] By www.jlr.org Published On :: 2020-05-05T13:30:26-07:00 Vitamin A aldehyde covalently bound to opsin protein is embedded in a phospholipid-rich membrane that supports photon absorption and phototransduction in photoreceptor cell outer segments. Following absorption of a photon, the 11-cis-retinal chromophore of visual pigment in photoreceptor cells isomerizes to all-trans-retinal. To maintain photosensitivity 11-cis-retinal must be replaced. At the same time, however, all-trans-retinal has to be handled so as to prevent nonspecific aldehyde activity. Some molecules of retinaldehyde upon release from opsin are efficiently reduced to retinol. Other molecules are released into the lipid phase of the disc membrane where they form a conjugate (N-retinylidene-PE, NRPE) through a Schiff base linkage with phosphatidylethanolamine (PE). The reversible formation of NRPE serves as a transient sink for retinaldehyde that is intended to return retinaldehyde to the visual cycle. However, if instead of hydrolyzing to PE and retinaldehyde, NRPE reacts with a second molecule of retinaldehyde a synthetic pathway is initiated that leads to the formation of multiple species of unwanted bisretinoid fluorophores. We report on recently identified members of the bisretinoid family some of which differ with respect to the acyl chains associated with the glycerol backbone. We discuss processing of the lipid moieties of these fluorophores in lysosomes of retinal pigment epithelial (RPE) cells, their fluorescence characters and new findings related to light and iron-associated oxidation of bisretinoids. Full Article
ino Retinoids in the visual cycle: Role of the retinal G protein-coupled receptor [Thematic Reviews] By www.jlr.org Published On :: 2020-06-03T16:30:29-07:00 Driven by the energy of a photon, the visual pigments in rod and cone photoreceptor cells isomerize 11-cis-retinal to the all-trans configuration. This photochemical reaction initiates the signal transduction pathway that eventually leads to the transmission of a visual signal to the brain and leaves the opsins insensitive to further light stimulation. For the eye to restore light sensitivity, opsins require recharging with 11-cis-retinal. This trans–cis back conversion is achieved through a series of enzymatic reactions composing the retinoid (visual) cycle. Although it is evident that the classical retinoid cycle is critical for vision, the existence of an adjunct pathway for 11-cis-retinal regeneration has been debated for many years. Retinal pigment epithelium (RPE)–retinal G protein-coupled receptor (RGR) has been identified previously as a mammalian retinaldehyde photoisomerase homologous to retinochrome found in invertebrates. Using pharmacological, genetic, and biochemical approaches, researchers have now established the physiological relevance of the RGR in 11-cis-retinal regeneration. The photoisomerase activity of RGR in the RPE and Müller glia explains how the eye can remain responsive in daylight. In this review, we will focus on retinoid metabolism in the eye and visual chromophore regeneration mediated by RGR. Full Article
ino Signaling roles of phosphoinositides in the retina [Thematic Reviews] By www.jlr.org Published On :: 2020-06-15T14:33:05-07:00 The field of phosphoinositide signaling has expanded significantly in recent years. Phosphoinositides (PIs) are universal signaling molecules that directly interact with membrane proteins or with cytosolic proteins containing domains that directly bind phosphoinositides and are recruited to cell membranes. Through the activities of PI kinases and PI phosphatases, seven distinct phosphoinositide lipid molecules are formed from the parent molecule phosphatidylinositol. PI signals regulate a wide range of cellular functions, including cytoskeletal assembly, membrane binding and fusion, ciliogenesis, vesicular transport, and signal transduction. Given the many excellent reviews on phosphoinositide kinases, phosphoinositide phosphatases, and PIs in general, in this review, we discuss recent studies and advances in PI lipid signaling in the retina. We specifically focus on PI lipids from vertebrate (e.g. bovine, rat, mice, toad, and zebrafish) and invertebrate (e.g. drosophila, horseshoe crab, and squid) retinas. We also discuss the importance of PIs revealed from animal models and human diseases, and methods to study PI levels both in vitro and in vivo. We propose that future studies should investigate the function and mechanism of activation of PI-modifying enzymes/phosphatases and further unravel PI regulation and function in the different cell types of the retina. Full Article
ino Lipid metabolism dysregulation in diabetic retinopathy [Thematic Reviews] By www.jlr.org Published On :: 2020-12-23T12:30:44-08:00 Lipid metabolic abnormalities have emerged as potential risk factors for the development and progression of diabetic complications, including diabetic retinopathy (DR). This review article provides an overview of the results of clinical trials evaluating the potential benefits of lipid lowering drugs, such as fibrates, omega 3 fatty acids, and statins, for the prevention and treatment of DR. Although several clinical trials demonstrated that treatment with fibrates leads to improvement of DR, there is a dissociation between the protective effects of fibrates in the retina, and the intended blood lipid classes, including plasma triglycerides, total cholesterol or HDL/LDL cholesterol ratio. Guided by these findings, plasma lipid and lipoprotein-independent mechanisms are addressed based on clinical, cell culture and animal model studies. Potential retinal-specific effects of fatty acids oxidation products, cholesterol, and ceramide, as well as lipid independent effects of PPAR alpha activation are summarized based on current literature. Overall, this review highlights promising potential of lipid-based treatment strategies further enhanced by the new knowledge of intra-retinal lipids and lipoproteins in DR. Full Article
ino {alpha}-Synuclein facilitates endocytosis by elevating the steady-state levels of phosphatidylinositol 4,5-bisphosphate [Membrane Biology] By www.jbc.org Published On :: 2020-12-25T00:06:30-08:00 α-Synuclein (α-Syn) is a protein implicated in the pathogenesis of Parkinson's disease (PD). It is an intrinsically disordered protein that binds acidic phospholipids. Growing evidence supports a role for α-Syn in membrane trafficking, including, mechanisms of endocytosis and exocytosis, although the exact role of α-Syn in these mechanisms is currently unclear. Here we investigate the associations of α-Syn with the acidic phosphoinositides (PIPs), phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and phosphatidylinositol 3,4-bisphosphate (PI(3,4)P2). Our results show that α-Syn colocalizes with PIP2 and the phosphorylated active form of the clathrin adaptor protein 2 (AP2) at clathrin-coated pits. Using endocytosis of transferrin as an indicator for clathrin-mediated endocytosis (CME), we find that α-Syn involvement in endocytosis is specifically mediated through PI(4,5)P2 levels on the plasma membrane. In accord with their effects on PI(4,5)P2 levels, the PD associated A30P, E46K, and A53T mutations in α-Syn further enhance CME in neuronal and nonneuronal cells. However, lysine to glutamic acid substitutions at the KTKEGV repeat domain of α-Syn, which interfere with phospholipid binding, are ineffective in enhancing CME. We further show that the rate of synaptic vesicle (SV) endocytosis is differentially affected by the α-Syn mutations and associates with their effects on PI(4,5)P2 levels, however, with the exception of the A30P mutation. This study provides evidence for a critical involvement of PIPs in α-Syn–mediated membrane trafficking. Full Article
ino High-dimensional Cytometry (ExCYT) and Mass Spectrometry of Myeloid Infiltrate in Clinically Localized Clear Cell Renal Cell Carcinoma Identifies Novel Potential Myeloid Targets for Immunotherapy [Research] By www.mcponline.org Published On :: 2020-11-01T00:05:37-07:00 Renal Cell Carcinoma (RCC) is one of the most commonly diagnosed cancers worldwide with research efforts dramatically improving understanding of the biology of the disease. To investigate the role of the immune system in treatment-naïve clear cell Renal Cell Carcinoma (ccRCC), we interrogated the immune infiltrate in patient-matched ccRCC tumor samples, benign normal adjacent tissue (NAT) and peripheral blood mononuclear cells (PBMCs isolated from whole blood, focusing our attention on the myeloid cell infiltrate. Using flow cytometric, MS, and ExCYT analysis, we discovered unique myeloid populations in PBMCs across patient samples. Furthermore, normal adjacent tissues and ccRCC tissues contained numerous myeloid populations with a unique signature for both tissues. Enrichment of the immune cell (CD45+) fraction and subsequent gene expression analysis revealed a number of myeloid-related genes that were differentially expressed. These data provide evidence, for the first time, of an immunosuppressive and pro-tumorigenic role of myeloid cells in early, clinically localized ccRCC. The identification of a number of immune proteins for therapeutic targeting provides a rationale for investigation into the potential efficacy of earlier intervention with single-agent or combination immunotherapy for ccRCC. Full Article
ino Kinome Profiling of Primary Endometrial Tumors Using Multiplexed Inhibitor Beads and Mass Spectrometry Identifies SRPK1 as Candidate Therapeutic Target [Research] By www.mcponline.org Published On :: 2020-12-01T00:05:33-08:00 Endometrial carcinoma (EC) is the most common gynecologic malignancy in the United States, with limited effective targeted therapies. Endometrial tumors exhibit frequent alterations in protein kinases, yet only a small fraction of the kinome has been therapeutically explored. To identify kinase therapeutic avenues for EC, we profiled the kinome of endometrial tumors and normal endometrial tissues using Multiplexed Inhibitor Beads and Mass Spectrometry (MIB-MS). Our proteomics analysis identified a network of kinases overexpressed in tumors, including Serine/Arginine-Rich Splicing Factor Kinase 1 (SRPK1). Immunohistochemical (IHC) analysis of endometrial tumors confirmed MIB-MS findings and showed SRPK1 protein levels were highly expressed in endometrioid and uterine serous cancer (USC) histological subtypes. Moreover, querying large-scale genomics studies of EC tumors revealed high expression of SRPK1 correlated with poor survival. Loss-of-function studies targeting SRPK1 in an established USC cell line demonstrated SRPK1 was integral for RNA splicing, as well as cell cycle progression and survival under nutrient deficient conditions. Profiling of USC cells identified a compensatory response to SRPK1 inhibition that involved EGFR and the up-regulation of IGF1R and downstream AKT signaling. Co-targeting SRPK1 and EGFR or IGF1R synergistically enhanced growth inhibition in serous and endometrioid cell lines, representing a promising combination therapy for EC. Full Article
ino Developments in Mass Spectrometry for Glycosaminoglycan Analysis: A Review [Review] By www.mcponline.org Published On :: 2020-09-16T09:35:59-07:00 This review covers recent developments in glycosaminoglycan (GAG) analysis via mass spectrometry (MS). GAGs participate in a variety of biological functions, including cellular communication, wound healing, and anticoagulation, and are important targets for structural characterization. GAGs exhibit a diverse range of structural features due to the variety of O- and N-sulfation modifications and uronic acid C-5 epimerization that can occur, making their analysis a challenging target. Mass spectrometry approaches to the structure assignment of GAGs have been widely investigated, and new methodologies remain the subject of development. Advances in sample preparation, tandem MS techniques (MS/MS), on-line separations and automated analysis software have advanced the field of GAG analysis. These recent developments have led to remarkable improvements in the precision and time efficiency for the structural characterization of GAGs. Full Article
ino Thermal proteome profiling in zebrafish reveals effects of napabucasin on retinoic acid metabolism [Research] By www.mcponline.org Published On :: 2020-10-28T08:35:46-07:00 Thermal proteome profiling (TPP) allows for the unbiased detection of drug – target protein engagements in vivo. Traditionally, one cell type is used for TPP studies, with the risk of missing important differentially expressed target proteins. The use of whole organisms would circumvent this problem. Zebrafish embryos are amenable to such an approach. Here, we used TPP on whole zebrafish embryo lysate to identify protein targets of napabucasin, a compound that may affect Signal transducer and activator of transcription 3 (Stat3) signaling through an ill-understood mechanism. In zebrafish embryos, napabucasin induced developmental defects consistent with inhibition of Stat3 signaling. TPP profiling showed no distinct shift in Stat3 upon napabucasin treatment, but effects were detected on the oxidoreductase, Pora, which might explain effects on Stat3 signaling. Interestingly, thermal stability of several aldehyde dehydrogenases (Aldhs) was affected. Moreover, napabucasin activated ALDH enzymatic activity in vitro. Aldhs have crucial roles in retinoic acid metabolism and functionally we validated napabucasin-mediated activation of the retinoic acid pathway in zebrafish in vivo. We conclude that TPP profiling in whole zebrafish embryo lysate is feasible and facilitates direct correlation of in vivo effects of small molecule drugs with their protein targets. Full Article
ino Proteome analysis reveals a significant host-specific response in Rhizobium leguminosarum bv viciae endosymbiotic cells [Research] By www.mcponline.org Published On :: 2020-11-19T08:37:14-08:00 The Rhizobium-legume symbiosis is a beneficial interaction in which the bacterium converts atmospheric nitrogen into ammonia and delivers it to the plant in exchange for carbon compounds. This symbiosis implies the adaptation of bacteria to live inside host plant cells. In this work we apply RP-LC-MS/MS and iTRAQ techniques to study the proteomic profile of endosymbiotic cells (bacteroids) induced by Rhizobium leguminosarum bv viciae strain UPM791 in legume nodules. Nitrogenase subunits, tricarboxylic acid cycle enzymes, and stress response proteins are amongst the most abundant from over one thousand rhizobial proteins identified in pea (Pisum sativum) bacteroids. Comparative analysis of bacteroids induced in pea and in lentil (Lens culinaris)nodules revealed the existence of a significant host-specific differential response affecting dozens of bacterial proteins, including stress-related proteins, transcriptional regulators, and proteins involved in the carbon and nitrogen metabolisms. A mutant affected in one of these proteins, homologous to a GntR-like transcriptional regulator, showed a symbiotic performance significantly impaired in symbiosis with pea, but not with lentil plants. Analysis of the proteomes of bacteroids isolated from both hosts also revealed the presence of different sets of plant-derived nodule-specific cysteine rich (NCR) peptides, indicating that the endosymbiotic bacteria find a host-specific cocktail of chemical stressors inside the nodule. By studying variations of the bacterial response to different plant cell environments we will be able to identify specific limitations imposed by the host that might give us clues for the improvement of rhizobial performance. Full Article
ino Proteomic analyses identify differentially expressed proteins and pathways between low-risk and high-risk subtypes of early-stage lung adenocarcinoma and their prognostic impacts [Research] By www.mcponline.org Published On :: 2020-11-30T14:35:18-08:00 The histopathological subtype of lung adenocarcinoma (LUAD) is closely associated with prognosis. Micropapillary or solid predominant LUAD tends to relapse after surgery at an early stage, whereas lepidic pattern shows a favorable outcome. However, the molecular mechanism underlying this phenomenon remains unknown. Here, we recruited 31 lepidic predominant LUADs (LR: low-risk subtype group) and 28 micropapillary or solid predominant LUADs (HR: high-risk subtype group). Tissues of these cases were obtained and label-free quantitative proteomic and bioinformatic analyses were performed. Additionally, prognostic impact of targeted proteins was validated using The Cancer Genome Atlas databases (n=492) and tissue microarrays composed of early-stage LUADs (n=228). A total of 192 differentially expressed proteins were identified between tumor tissues of LR and HR and three clusters were identified via hierarchical clustering excluding eight proteins. Cluster 1 (65 proteins) showed a sequential decrease in expression from normal tissues to tumor tissues of LR and then to HR and was predominantly enriched in pathways such as tyrosine metabolism and ECM-receptor interaction, and increased matched mRNA expression of 18 proteins from this cluster predicted favorable prognosis. Cluster 2 (70 proteins) demonstrated a sequential increase in expression from normal tissues to tumor tissues of LR and then to HR and was mainly enriched in pathways such as extracellular organization, DNA replication and cell cycle, and high matched mRNA expression of 25 proteins indicated poor prognosis. Cluster 3 (49 proteins) showed high expression only in LR, with high matched mRNA expression of 20 proteins in this cluster indicating favorable prognosis. Furthermore, high expression of ERO1A and FEN1 at protein level predicted poor prognosis in early-stage LUAD, supporting the mRNA results. In conclusion, we discovered key differentially expressed proteins and pathways between low-risk and high-risk subtypes of early-stage LUAD. Some of these proteins could serve as potential biomarkers in prognostic evaluation. Full Article
ino Optimized incorporation of an unnatural fluorescent amino acid affords measurement of conformational dynamics governing high-fidelity DNA replication [DNA and Chromosomes] By www.jbc.org Published On :: 2020-12-11T00:06:21-08:00 DNA polymerase from bacteriophage T7 undergoes large, substrate-induced conformational changes that are thought to account for high replication fidelity, but prior studies were adversely affected by mutations required to construct a Cys-lite variant needed for site-specific fluorescence labeling. Here we have optimized the direct incorporation of a fluorescent un-natural amino acid, (7-hydroxy-4-coumarin-yl)-ethylglycine, using orthogonal amber suppression machinery in Escherichia coli. MS methods verify that the unnatural amino acid is only incorporated at one position with minimal background. We show that the single fluorophore provides a signal to detect nucleotide-induced conformational changes through equilibrium and stopped-flow kinetic measurements of correct nucleotide binding and incorporation. Pre-steady-state chemical quench methods show that the kinetics and fidelity of DNA replication catalyzed by the labeled enzyme are largely unaffected by the unnatural amino acid. These advances enable rigorous analysis to establish the kinetic and mechanistic basis for high-fidelity DNA replication. Full Article
ino Phillies, Sean Rodriguez agree to Minors deal By mlb.mlb.com Published On :: Fri, 8 Feb 2019 13:26:36 EDT The Phillies continued to bolster their infield depth on Friday by signing versatile veteran Sean Rodriguez to a Minor League contract with an invite to big league Spring Training, MLB.com has learned. Full Article
