ida Google’s ‘Where to Vote’ Search Result Reflects Quirk of Candidate Surname, Not Bias By www.factcheck.org Published On :: Wed, 06 Nov 2024 21:56:25 +0000 Social media users alleged bias against former President Donald Trump when a Google search on Election Day for “where to vote” returned an interactive map to find a person’s polling station when including the word “Harris” but not “Trump.” The reason is because “Harris” is a county in Texas, whereas “Trump” is not a location. The post Google’s ‘Where to Vote’ Search Result Reflects Quirk of Candidate Surname, Not Bias appeared first on FactCheck.org. Full Article Debunking Viral Claims FactCheck Posts 2024 elections Presidential Election 2024
ida Friday Girls From Russian Province By englishrussia.com Published On :: Fri, 18 Feb 2022 09:58:02 +0000 The post Friday Girls From Russian Province appeared first on English Russia. Full Article Photos Russian People girls women
ida Frida's Flowers - 1 Year Crochet Project - So Much Fun! By beadlust.blogspot.com Published On :: Mon, 21 Aug 2017 10:02:00 +0000 It All Started with Hexie-lust! Look back to early 2012, when my dear friend, Christy, started covering little hexagon-shaped bits of card stock with fabric, making stacks of 6 ready to sew together as petals for a hexie flower. While I was working on a beading project, she was making these utterly irresistible stacks of hexies, until finally I succumbed to the temptation, and joined her. Here is Christy holding a whole bag of covered hexie shapes, made for the pathway around her hexie flowers. And here I am holding my hexie quilt, showing the final seam of hand stitching needed to complete the quilt top for my hexie quilt, Grandma's Flower Garden. You'd think that hand-stitching and quilting over 4,000 hexies would be enough for any sane person, right? Guess I'm not sane, because in 2015 I started another hexie quilt, or as it turns out, a triptych of wall quilts, for which I've completed 733 hexie flowers (which requires 5,131 individual hexies)! Getting Hooked on Crochet OK.... so now, I'm finished with hexies, right? You guessed it! The answer is, "NO WAY." And once again it's Christy's "fault." With her hexie quilt on the back burner for a while, she moved into a crochet phase, making lovely afghans and shawls. She, along with Sabine, my friend in Germany, got me hooked on crochet (pun intended). This is the first shawl I made late in 2015, learning and getting comfortable with the hook. Then.... the big bang happened... the crochet-hexie connection!!! This is it. I saw this picture on Huib Petersen's Facebook page, fell bonkers in love, messaged him to find out about it, and learned that there is a pattern for the flowers online. Click on the above photo of Huib's flowers to see it enlarged... Wouldn't you be a bit inclined to go bonkers over it too??? On June 12, 2016, just one day after seeing his crocheted hexie flowers on Facebook, Christy and I were in Island Wools, our local yarn shop, buying DK-weight, cotton yarn in a dozen colors, ready to begin our own stacks of crocheted hexie flowers! On a whim, right there in the yarn shop, we decided to keep what we were doing a secret... not to show or tell anybody about our project until we finished our afghans... no blogging or posting on Facebook about it. We didn't even tell Libby or Julie at the yarn shop why we kept ordering more cotton yarn. Our secret-keeping made it all the more fun! Original Crochet-Along, Frida's Flowers According to Huib, the instructions for his flowers came from a Stylecraft, Crochet-Along, called Frida's Flowers, staring an original pattern by Jane Crowfoot. This is a photo from the instructions, showing the finished afghan, which includes several identical flowers in each of 2 simple and 5 complex designs. All of the designs are are multi-colored and textural, with raised flower parts, enough to make us drool! Bored with making 6 identical flowers - Colors calling us! Our plan was to get together at my house every Sunday afternoon to crochet hexie flowers, each of us completing enough flowers to make an afghan. In a little over a year of working 4-6 hours nearly every Sunday and some Tuesday evenings as well, we each had completed 39 flower blocks and 6 half-flower blocks, and were ready to crochet them together. Ooops... I'm getting ahead of myself with this story. We began with this block, called Rosa, which was the 3rd block in the overall design. (Blocks 1 and 2 are the more simple ones with a small central bud and plain background). This one is the easiest of the full flower designs. But, for both of us, it was difficult, as there were several stitches we didn't know. Thanks to Youtube videos, we were able to learn them. However, after making two flowers each in the pattern colors, partially out of boredom and partly because of the influence of Huib's multi-color, no-two-the-same flowers, we decided to pick our own colors, making only a pair in each colorway. This, of course, caused us both to buy a lot more colors of yarn... oh for fun! After making 6 each of Blocks 3 and 4, mine looked like this. Christy's color choices are different... enough different that our finished afghans may look like sisters, but definitely not like identical twins. I started looking at flower catalogs to find new color combinations... and both of us were buying yarn like crazy. Some brands have more that 50 color choices in DK-weight cotton. I admit to spending over two hundred dollars on yarn all-in-all, with some remainders for future projects. Never mind the cost... I adore all the colors. Designing Our Afghans Early in the process, both Christy and I decided we wanted to make something more like Huib's, with a random or nearly random placement of the blocks. Plus, we wanted it to be a bit bigger than the 31-block original design. Also, we didn't want to include any of the more simple blocks, except as modified half-blocks for the sides. A year went by, with the two of us continually challenged, thoroughly enjoying the process of making our blocks. Then it was time to lay them all out! Almost at once, it was clear that the flowers needed more space, more black around each one to set them off. So, before crocheting them together, we bought more skeins of black yarn, and added a row of double crochet around each of the blocks. This also would add a bit more to the size of the afghans, making them large enough to cover both arms and legs while watching a good movie on a winter's evening. Here is my finished arrangement, the hexie flower blocks crocheted together with a slip-stitch, awaiting a border. Although the original design included a border that would have worked OK, by then I was flying solo, wanting a border I could call my own. After some experimentation, trial-and-error, crochet and un-ravel attempts, this is my final border invention, which includes the "popcorn" stitch, central to many of the flowers. It was challenging to figure out how to crochet the increases and decreases necessary for the zig-zag edges on the sides, and still keep it flat. Again, trial-and-error was part of the process. Here is my almost-finished afghan, my own version of Frida's Flowers, showing the size! Entering at the San Juan County Fair As we neared completion, we faced a moral dilemma, a difficult decision. We both enjoy submitting entries at our local San Juan County Fair each year, especially in the Fiber and Textile Arts Divisions. These entries are judged and eligible to win ribbons and cash prizes. In previous years, there haven't been many crocheted items entered, nothing that has won any of the top awards. We figured our afghans could be "game changers," that they had a chance of winning. But we didn't like the idea of being in competition with each other for the top awards, the Best of Class and the Best of Show. If we both entered, neither quilt would win a top award, or one would win and the other wouldn't (which might be the worst outcome). So, after some heartfelt discussions, we decided I would enter mine this year, and she would delay finishing hers until later so that it would be eligible to enter next year. Here's what happened... Best of Class and Viewer's Choice for me in 2017!!!! And hopefully, the same will happen next year for Christy's version. Twelve months from now, I know for sure all the attendees will have forgotten my quilt, and will love seeing Christy's flowers, just as they did mine this year! Full Article afghan Awards crochet design Frida's Flowers grandma's flower garden hexie flowers hexies Huib Petersen Jane Crowfoot
ida University Holiday - Thanksgiving By www.princeton.edu Published On :: Thu, 28 Nov 2024 00:00:00 -0500 Full Article
ida Happy Thanksgiving and Black Friday Deal By www.rssground.com Published On :: Tue, 23 Nov 2021 18:44:18 +0000 We are taking this opportunity to say Thank You! We can’t express how much we appreciate you being a part of RSS Ground family. We hope our unique tools will continue to be useful to you for years to come. We usually never do this, but this time we would like to show our token […] The post Happy Thanksgiving and Black Friday Deal appeared first on RSSground.com. Full Article RSS Ground News Black Friday Deal discount Happy Holidays
ida The perfect holiday in Val d’Isère, Britain's favourite French ski resort By www.telegraph.co.uk Published On :: Wed, 04 Sep 2024 17:00:00 GMT Full Article structure:travel/ski topics:places/french-alps structure:travel structure:travel/ski/resort-guides topics:places/val-d-isere storytype:standard structure:eg-general structure:us-content
ida The perfect ski holiday in Canada's oldest national park By www.telegraph.co.uk Published On :: Sat, 07 Sep 2024 17:00:00 GMT Full Article structure:travel/ski structure:travel structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:better-life/better-life-evergreen structure:better-life/better-life-guide structure:travel/destination-guides-100
ida The perfect ski holiday in Courchevel, one of France’s most glamorous resorts By www.telegraph.co.uk Published On :: Tue, 10 Sep 2024 19:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/courchevel structure:travel/ski/resort-guides storytype:standard topics:places/france structure:eg-general structure:us-content structure:better-life/better-life-evergreen structure:better-life/better-life-guide
ida The perfect ski holiday in Austria's most exclusive resort By www.telegraph.co.uk Published On :: Thu, 19 Sep 2024 14:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/lech structure:travel/ski/resort-guides storytype:standard topics:places/austria
ida An expert guide to ski holidays in Italy’s most quintessential ski resort By www.telegraph.co.uk Published On :: Sat, 21 Sep 2024 12:00:00 GMT Full Article structure:travel/ski structure:travel/ski/resort-guides structure:travel topics:places/italy storytype:standard structure:eg-general structure:better-life/better-life-evergreen structure:better-life/better-life-guide
ida The perfect ski holiday in Chamonix, the most legendary resort in France By www.telegraph.co.uk Published On :: Tue, 24 Sep 2024 17:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/chamonix structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:us-content structure:better-life/better-life-evergreen structure:better-life/better-life-guide structure:travel/destination-guides-100 topics:places/europe topics:places/france
ida The perfect ski holiday in Telegraph reader's favourite Austrian ski resort By www.telegraph.co.uk Published On :: Fri, 27 Sep 2024 12:00:00 GMT Full Article topics:places/st-anton structure:travel/ski/resort-guides structure:travel/ski structure:eg-general storytype:standard topics:places/austria topics:places/europe structure:better-life/better-life-guide structure:better-life/better-life-evergreen
ida The perfect holiday in Verbier – Switzerland’s coolest ski resort By www.telegraph.co.uk Published On :: Tue, 01 Oct 2024 06:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/verbier structure:travel/ski/resort-guides storytype:standard topics:places/switzerland structure:us-content structure:better-life/better-life-evergreen structure:better-life/better-life-guide topics:places/europe
ida The perfect ski holiday in Cortina d'Ampezzo, Italy's chicest mountain resort By www.telegraph.co.uk Published On :: Fri, 04 Oct 2024 13:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/cortina topics:places/italy structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:better-life/better-life-evergreen topics:places/dolomites structure:better-life/better-life-guide structure:travel/destination-guides-100 topics:places/europe
ida The perfect ski holiday in St Moritz, the birthplace of winter sports By www.telegraph.co.uk Published On :: Mon, 14 Oct 2024 16:00:00 GMT Full Article structure:travel/ski structure:travel structure:travel/ski/resort-guides topics:places/st-moritz topics:places/switzerland storytype:standard structure:eg-general structure:better-life/better-life-evergreen structure:better-life/better-life-guide topics:places/europe structure:travel/destination-guides-100
ida The perfect ski holiday in Morzine By www.telegraph.co.uk Published On :: Sun, 20 Oct 2024 09:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/morzine structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:better-life/better-life-evergreen structure:travel/destination-guides-100 topics:places/france topics:places/europe
ida An expert guide to Avoriaz, the French resort designed for snow-sure ski holidays By www.telegraph.co.uk Published On :: Mon, 28 Oct 2024 17:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/avoriaz structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:better-life/better-life-guide structure:better-life/better-life-evergreen topics:places/france topics:places/europe
ida The perfect ski holiday in Méribel, the beating heart of France’s biggest ski area By www.telegraph.co.uk Published On :: Wed, 30 Oct 2024 13:00:00 GMT Full Article structure:travel/ski structure:travel structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:us-content structure:better-life/better-life-evergreen structure:better-life/better-life-guide structure:travel/destination-guides-100 topics:places/europe topics:places/france
ida The perfect ski holiday in Courmayeur, Italy’s stylish answer to Chamonix By www.telegraph.co.uk Published On :: Thu, 31 Oct 2024 16:00:00 GMT Full Article structure:travel/ski structure:travel structure:travel/ski/resort-guides storytype:standard structure:eg-general topics:places/italy structure:better-life/better-life-guide structure:better-life/better-life-evergreen topics:places/europe structure:travel/destination-guides-100
ida The perfect ski holiday in Whistler, Canada’s leading winter resort By www.telegraph.co.uk Published On :: Thu, 07 Nov 2024 18:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/whistler structure:travel/ski/resort-guides storytype:standard topics:places/canada structure:eg-general structure:travel/destination-guides-100 structure:better-life/better-life-evergreen structure:better-life/better-life-guide
ida The perfect holiday in underrated Lanzarote By www.telegraph.co.uk Published On :: Fri, 08 Nov 2024 17:00:00 GMT Full Article structure:travel/restaurants structure:travel/nightlife structure:travel/hotels structure:travel/destination-guides-100 structure:travel structure:travel/attractions storytype:standard default:eg_general structure:better-life/better-life-evergreen structure:better-life/better-life-guide topics:places/europe topics:places/lanzarote
ida The perfect ski holiday in Les Arcs By www.telegraph.co.uk Published On :: Sun, 10 Nov 2024 14:00:00 GMT Full Article structure:travel/ski structure:travel topics:places/les-arcs structure:travel/ski/resort-guides storytype:standard structure:eg-general structure:travel/destination-guides-100 structure:better-life/better-life-guide structure:better-life/better-life-evergreen topics:places/europe topics:places/france
ida How to spend the perfect holiday in Tenerife By www.telegraph.co.uk Published On :: Mon, 11 Nov 2024 20:00:00 GMT Full Article structure:travel/restaurants structure:travel/nightlife structure:travel/hotels structure:travel/destination-guides-100 structure:travel structure:travel/attractions storytype:standard structure:better-life/better-life-evergreen structure:better-life/better-life-guide
ida Video: Friday Fails #332 By www.pinkbike.com Published On :: Fri, 8 Nov 2024 00:00:00 PST Pile ups and loop outs...This week's Friday Fails has it all!( Comments: 46 ) Full Article
ida Video: A Summer Bike Holiday with Friends in Morzine By www.pinkbike.com Published On :: Sun, 10 Nov 2024 00:00:00 PST Ride, eat, party, sleep, repeat.( Comments: 49 ) Full Article
ida The Pinkbike Holiday Gift Guide: Top Gifts for All the MTBers in Your Life By www.pinkbike.com Published On :: Tue, 12 Nov 2024 00:00:00 PST We've pulled together an eclectic mix of mountain bike-related gift ideas for all the riders on your list.( Photos: 12, Comments: 49 ) Full Article
ida A day at a Florida manatee hospital By www.pbs.org Published On :: Tue, 29 Mar 2022 01:09:00 +0000 As Florida’s seagrass beds die off, manatees are starving. Can the seagrass–and the manatees–make a comeback? Full Article
ida Best cities to visit in Florida, from Miami to Jacksonville By www.independent.co.uk Published On :: 2024-11-12T09:57:51+00:00 The most popular US state for British tourists has plenty more to offer than Disney – experts Susan Veness and Simon Veness share eight destinations for a fun-packed city break Full Article Florida USA North America Travel
ida My American family couldn’t visit me in London, so we went to ‘London’ in Florida instead By www.independent.co.uk Published On :: 2024-11-12T12:00:46+00:00 Having lived in London for four years, American Kassondra Cloos thought a trip see her adopted city as imagined by the Wizarding World of Harry Potter in Orlando would be fairly pointless – but instead she finds a home from home that’s suited to even grown-up kids Full Article Orlando Florida USA North America Travel
ida Half-term holidays give London Stansted busiest passenger day in airport history By www.independent.co.uk Published On :: 2024-11-12T13:59:01+00:00 Over 100,000 passengers passed through the airport on 27 October Full Article News & Advice Travel
ida Bali flights cancelled after volcanic eruption near holiday spot creates dangerous ash cloud By www.independent.co.uk Published On :: 2024-11-13T07:49:43+00:00 Over a dozen flights cancelled or delayed between Bali and Australia Full Article News & Advice Travel
ida Vida em lua de Urano? As revelações de nova pesquisa sobre condições do planeta By www.bbc.com Published On :: Tue, 12 Nov 2024 11:26:12 GMT Dados enviados pela Voyager 2 em 1986 mostravam que as luas de Urano eram completamente diferente do resto do Sistema Solar Full Article
ida Florida Lawmakers Want Accessible Liquefied Natural gas By shark-tank.com Published On :: Wed, 20 Mar 2019 19:14:54 +0000 “This bill makes American liquefied natural gas accessible and cost-effective to regional allies." The post Florida Lawmakers Want Accessible Liquefied Natural gas appeared first on Shark Tank. Full Article Featured florida Florida Politics Jacksonville venezuela
ida FEMA Worker Being Scapegoated for Denying Hurricane Aid to Trump Supporters Says It was Agency Policy in Florida and The Carolinas By sonsoflibertymedia.com Published On :: Tue, 12 Nov 2024 19:15:36 +0000 The FEMA official who was just fired for telling workers to avoid homes impacted by the hurricanes in Florida if they had Trump signs says that it was not “isolated” and that FEMA workers were instructed to do it in the Carolinas too. (X) The American people must get justice. Indict and convict. So, apparently this … Full Article Commentary Videos
ida Yale psychiatrist says you should avoid your Trump-voting relatives during the holidays By libertyunyielding.com Published On :: Tue, 12 Nov 2024 03:40:58 +0000 Yale medical school’s chief psychiatry resident told a TV host that people should avoid their Trump-voting relatives on Thanksgiving and Christmas. While on MSNBC with host Joy Reid, Amanda Calhoun said if you have relatives who voted for Trump, “it’s completely fine to not be around those people and to tell them why.” Calhoun suggested […] The post Yale psychiatrist says you should avoid your Trump-voting relatives during the holidays appeared first on Liberty Unyielding. Full Article Education
ida Why Kamala Harris lost: A flawed candidate or doomed campaign? By www.bbc.com Published On :: Thu, 07 Nov 2024 01:13:37 GMT The vice-president failed to make the case that she would be different from her boss, President Joe Biden. Full Article
ida velocityconf: Free webcast from our friends at @citrix 5/29 http://t.co/IOeY4U0wUP Learn to consolidate 40 load balancers and ADCs into single platform By twitter.com Published On :: Tue, 21 May 2013 15:01:58 +0000 velocityconf: Free webcast from our friends at @citrix 5/29 http://t.co/IOeY4U0wUP Learn to consolidate 40 load balancers and ADCs into single platform Full Article
ida velocityconf: New #velocityconf CA program preview is up: http://t.co/rKjf91RXdD @ariyahidayat on End-to-End JS Quality Analysis. By twitter.com Published On :: Thu, 23 May 2013 17:01:13 +0000 velocityconf: New #velocityconf CA program preview is up: http://t.co/rKjf91RXdD @ariyahidayat on End-to-End JS Quality Analysis. Full Article
ida Ergatta Rower Black Friday sale: Save up to $850 By www.businessinsider.com Published On :: Tue, 12 Nov 2024 18:19:57 +0000 Ergatta's Black Friday deals are live now, giving shoppers a chance to save up to $850 on its smart rowers, including free shipping. Full Article Deals (Reviews) Reviews Health (Reviews) black-friday insider-reviews ergatta
ida Best early Black Friday MacBook deals: Lowest-ever prices on top models By www.businessinsider.com Published On :: Tue, 12 Nov 2024 18:50:38 +0000 The best early Black Friday MacBook deals include all-time low prices on the latest MacBook Air models. Full Article Deals (Reviews) Reviews Black Friday & Cyber Monday (Reviews) Tech (Reviews) black-friday cyber-monday insider-picks ip-deals deals insider-reviews product-card black-friday-2024
ida The best Amazon Black Friday 2024 deals By www.businessinsider.com Published On :: Tue, 12 Nov 2024 20:51:04 +0000 Black Friday is weeks away, but Amazon already has several great deals you can find now. Here are the best Amazon early Black Friday deals. Full Article Deals (Reviews) Reviews Black Friday & Cyber Monday (Reviews) amazon black-friday insider-reviews
ida A brief history of America's love affair with fluoridated water — and why it's now up for debate By www.businessinsider.com Published On :: Tue, 12 Nov 2024 21:23:05 +0000 Too much fluoride can make your teeth brown, but getting a little bit is a dentist's dream. Here's the complete history of fluoridated water. Full Article Health fluoride public-health tap-water rfk-jr donald-trump
ida News24 | Africa has other friends if Trump won't work with it - Kenya's AU candidate By www.news24.com Published On :: Saturday Nov 09 2024 18:36:48 Kenya's candidate for the top job at the African Union said on Saturday that if US president-elect Donald Trump does not want to work with Africa, the continent has "other friends". Full Article
ida Joe Rogan Officially Backs A Candidate… Could Be The Biggest Endorsement Yet By clashdaily.com Published On :: Tue, 05 Nov 2024 10:23:15 +0000 There is a reason Joe Rogan became the world's largest podcaster. He doesn't take his audience -- or his guests -- for granted. He wants MEANINGFUL conversations. The post Joe Rogan Officially Backs A Candidate… Could Be The Biggest Endorsement Yet appeared first on Clash Daily. Full Article News Clash
ida Validation is a mirage By signalvnoise.com Published On :: Tue, 22 Dec 2020 17:07:56 +0000 Spend enough time talking with entrepreneurs, product people, designers, and anyone charged with proving something, and you’ll bump into questions about validation. “How do you validate if it’s going to work?”“How do you know if people will buy it to not?”“How do you validate product market fit?”“How do you validate if a feature is worth… keep reading Full Article Uncategorized
ida Sport | Here are the 3 candidates in line to replace Bill Beaumont as World Rugby chairman By www.news24.com Published On :: Wednesday Nov 13 2024 09:31:17 The race to succeed Bill Beaumont as chairman of World Rugby comes to a conclusion in a vote in Dublin at the governing body's headquarters on Thursday. Full Article
ida Investigation: Waste of the Day – Florida Hospitals Spent $566 Million Caring for Illegal Immigrants By deneenborelli.com Published On :: Tue, 01 Oct 2024 20:25:26 +0000 Investigation by Jeremy Portnoy originally published by RealClearInvestigations and RealClearWire Topline: Florida hospitals spent $566 million providing care for illegal immigrants in the last six months of 2023, according to a recent report from the state’s Agency for Healthcare Administration. Key facts: Gov. Ron DeSantis passed a law in May 2023 instructing all hospitals that … Full Article Commentaries Government Policies News
ida High affinity binding of SARS-CoV-2 spike protein enhances ACE2 carboxypeptidase activity [Molecular Bases of Disease] By www.jbc.org Published On :: 2020-12-25T00:06:31-08:00 The novel severe acute respiratory syndrome coronavirus (SARS-CoV-2) has emerged to a pandemic and caused global public health crisis. Human angiotensin-converting enzyme 2(ACE2) was identified as the entry receptor for SARS-CoV-2. As a carboxypeptidase, ACE2 cleaves many biological substrates besides angiotensin II to control vasodilatation and vascular permeability. Given the nanomolar high affinity between ACE2 and SARS-CoV-2 spike protein, we investigated how this interaction would affect the enzymatic activity of ACE2. Surprisingly, SARS-CoV-2 trimeric spike protein increased ACE2 proteolytic activity ∼3-10 fold against model peptide substrates, such as caspase-1 substrate and Bradykinin-analog. The enhancement in ACE2 enzymatic function was mediated by the binding of SARS-CoV-2 spike RBD domain. These results highlighted the potential for SARS-CoV-2 infection to enhance ACE2 activity, which may be relevant to the cardiovascular symptoms associated with COVID-19. Full Article
ida Reply to Sysel et al.: Comment on the importance of using nitric oxide gas in the synthesis of nitrosylcobalamin and ICH-validated methods to assess purity and stability [Letters to the Editor] By www.jbc.org Published On :: 2020-10-23T00:06:00-07:00 In their comment (1) on our publication (2), the authors make two points: (i) they raise concerns about the possible effect of residual NONOate in our study, and (ii) they promote nitrosylcobalamin (NOCbl) supplied by their own company. Both points lack merit for the following reasons. The authors make the astonishing claim that the spectra of nitric oxide (NO•) and cobalamins overlap. Unlike NO•, cobalamin absorbs in the visible region, permitting unequivocal spectral assignment of NOCbl as reported (3). We demonstrated that whereas NOCbl is highly unstable in solution, it is stabilized by the B12 trafficking protein CblC. So even if present, residual NONOate (which is unstable at neutral pH and is removed during the work-up (3)) could not account for the observed difference.The authors then misrepresent our synthetic method, claiming that anaerobic conditions were used to generate nitrocobalamin (NO2Cbl), which results in the transient formation of NOCbl. We synthesized NO2Cbl aerobically using nitrite as described (4); NOCbl is not an intermediate in this ligand exchange reaction. The aerobic instability of NOCbl has been rigorously described by inorganic chemists (3, 5) and raises obvious questions about its purported biological effects as exemplified by the authors' own 2003 JBC publication, which was later withdrawn.As to promoting NOCbl from their company, the authors refer to a synthetic route from a mixture of NO• gas and aquocobalamin. The authors' method (6) has been described as “dubious” by chemists (5). Whereas DEAE NONOate used in our method is widely known as an NO• donor,... Full Article
ida Comment on the importance of using nitric oxide gas in the synthesis of nitrosylcobalamin and ICH-validated methods to assess purity and stability [Letters to the Editor] By www.jbc.org Published On :: 2020-10-23T00:06:00-07:00 After a thorough read of this paper (1), we wish to clarify that the authors' anaerobic method of synthesis for the production of nitrocobalamin results in the transient formation of nitrosylcobalamin, an unstable intermediate upon exposure to air. We concur that the authors' method results in the production of nitrocobalamin based on the UV-visible data as shown. The authors' adapted anaerobic method consists of mixing hydroxocobalamin hydrochloride with diethylamine NONOate diethylammonium salt in aqueous solution. Of concern, the UV spectrum of nitric oxide overlaps that of all cobalamin species under anaerobic conditions, making any assignments of the binding of nitric oxide to hydroxocobalamin suspect (2). Additionally, the use of acetone to precipitate the authors' product causes precipitation of diethylamine NONOate, resulting in an impure product. As a result, its utility for drawing experimental conclusions is faulty.The product from the authors' anaerobic synthetic method has not been assessed for purity, and the synthetic method itself has not been validated using a stability-indicating method as required by the International Conference on Harmonization (ICH) (ICH Q2B, Validation of Analytical Procedures) methodology, which is a hallmark for analytical characterization. Our nitrosylcobalamin synthesis involves reacting nitric oxide gas with hydroxocobalamin acetate as a heterogeneous mixture in a non-electron-donating solvent followed by rotary evaporation. Our nitrosylcobalamin product is stable in air, releases nitric oxide gas in situ (3), and meets ICH stability guidelines (4). Additionally, our nitrosylcobalamin product demonstrates biological activity, which has not been observed for nitrocobalamin (3, 5). Full Article