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This was a really good episode. Lots of performances, lots of judgment. It went mostly the way I wanted it. I’m not going to say a whole lot about the people who didn’t make it through except that I was sad for Lauren Gray, and if Adam Brock goes, I’m going to be even more sad. I was crying a little already, watching him talk about how much singing meant to him, not that anyone else wouldn’t say that. Although saying he can’t wait for his daughter to say her dad was on Idol, lets be realistic, Idol better not still be around.
And now onto more pressing, already determined things. The person I was super ecstatic to see go, Cowboy Guy! What sweet sweet vindication that the douchebag country boy didn’t make it through. Nothing against country boys because I loved Scotty early on, but I hate this guys attitude, his cockiness, his overall douche bagginess. His swan song choice was great too, because I would love to see him fall into a burning ring of fire. Even the fire lake thing around him and the dead tree thing behind him was giving me a nice visual. Something tells me he wanted to just totally kirk out on them, I’m glad he stayed strong and didn’t, but I’m totally happy to see him go. Ryan, get your character liking in order because I couldn’t be happier to see his horrible character go!
I’ll start with the girls because while I like them, the boys are my stars this season.
Jen Hirsh – I’ve really liked Jen since I first saw her on Group Night with Reed Grimm, although I’m seeing why they didn’t air her audition performance. As for her final performance, I definitely didn’t think this was her best, but I do think that of the girls I’ve seen, she totally deserves to be through. Plus she’s first of the night and the producers don’t want to go ruining the show for me this early. I do worry for future auditions, that she’s got a case of the Jacob Lusk’s. You know what I mean. The endless, don’t know when to stop the note issues.
Haley Johnson – I don’t think we saw her in the original auditions but I will say that I can’t forget her group night performance. She really stuck out. I guess they didn’t want to show her final performance. I’m fine with leaving her group performance as the last thing I remember her singing, it was good enough for me!
Elise Testone – And in the same group as Haley Johnson, there was Elise Testone, who stood out much more than Haley, to me. I thought her final audition of This is a Mans World was lovely. She looks a little like Gaga in the face. I liked when she said that when she sings, she’s trying to convey every emotion she has – that’s the kind of performance that I want to see.
Erika Van Pelt – You Erica, I remember your original audition, I remember the other performances you had, I haven’t seen you sing something poorly yet, and I love the raspy deep Adeleness of your voice. Lord knows the Idol Producers would LOVE to discover the next Adele. The only thing she needs to work on is clothing choice. Hopefully the Idol stylists don’t muck things up.
I’d decided by this point in the show dramatic pause this season has resulted in a safety. By the end of the night I totally regretted saying that though, because we don’t know the status of Adam Brock…
Chelsea Sorrell – I really thought this girl would get sent home. We haven’t really seen her in the auditions except for little snippets. Maybe it was because of that herpe thing on her lip? It was gone by her final meeting with the judges but I noticed it. I’m just not entirely sure who this chick is. One last thing, does anyone else think, looking at her face that she kind of looks like Jennifer Tilly a little bit? a young Jennifer Tilly? I drew up a little comparison for you.
Bailey Brown – She has grown so much since her first days on Idol when those Jersey idiots ruined everything for her. I was really worried after they let Chelsea through that they wouldn’t let Bailey, and more nervous when they mentioned her consistency problems. I hate when they ask “How did you feel about your performance” because in my head that means you’re in trouble. Luckily they did it to so many last night, by the third of fourth I knew she was most likely safe. I agree that her last performance wasn’t the best but her first performance still sticks out in my head and I hope that’s the person she brings to the big stage!
Jessica Sanchez – Can I just throw it out there that I hate this girl? I’d like to say through no fault of her own but I really don’t’ know if I can. When she was saying she wanted to do well because “my family has spent so much money on me” or how about “My mom is unemployed because of my music career”. No mother should choose to be unemployed (especially with that large family you seem to have) because your daughter has a dream. You need to put food on your table. Not hope your daughter makes it so she can put food on your table. I also didn’t really like her previous performances, even though I know there are others out there who’ve already fallen in love with her. I had her in my “I’m totally fine if she leaves because she isn’t that good” column. Yeah, I’ve got one of those. What of it? Then I had to eat crow because her final performance was amazing, and promptly moved her over to my “safe” column.
Brielle Von Hugel – I don’t like her attitude. I don’t like her face. I don’t like her singing. I agree with Seacrest that she’s got moxie, but she also has a really gnarly bad sense of style (your tan tank top/undershirt looked so weird) I also don’t like her Toddlers & Tiara’s mother. Maybe it’s their overly NY accents. They are an automatic turn off for me. I wanted her gone like I wanted Cowboy Guy gone. If it had to be one or the other though, I’ll take Cowboy Guy gone, any day of the week!
Ok so that was all for the girls. Of this group, I think Erica Van Pelt and Baylie Brown are my favorites, but I think Jessica Sanchez will do the best. I’d be happy to be proven wrong though 🙂 We also still have the following girls left to see (that I’m aware of):
I bolded the ladies on the list I’d like to see make it through. And with that we move onto the boys.
Creighton Fraker – I was really surpised that this guy made it through. I knew he would when we got to learn a little more about Mr. Fraker. For example, his love of hideous glasses. Or that his birth father was the lead singer of Flotsam and Jetsam. Who knew that was a band? Not me, that’s for sure. I’m only going to bring up his final performance because of a second Billy Joel song I heard later in the evening that was infinately better. I’m befuddled that he was kept after hearing both Joel performances, to be honest. Maybe it just wasn’t the best song choice for him. We shall see.
Joshua Ledet – I know this kid is a judges favorite. They swoon upon sight of him. Seeing more of his performances tonight only made me like him less. Am I missing something that others see? I guess everyone has their own opinion but even my favorite and most respected Idol reviewers, like Michael Slezak. At least even dear Mr. Slezak aknowledges he could also have a case of the aforementioned Lusk Syndrome, or as he calls it, The Lusky Stank. I guess I can just go to my go to when I don’t like people – he just doesn’t do anything for me. I know he loves Jesus and can seemingly rock a gospel song, but all the “Aaaaamens” between Randy and Jennifer, I’m already over him.
Reed Grimm – Talk about someone I’m totally NOT over! I have loved every song choice he’s ever made. One small thing that bothers me about him is that he definitely has a small case of the crazy eyes. I’m gonna pretend it’s in a good way. I just really like his whole schtick. Do what Casey James couldn’t do and crack the Top 5? It would’ve been a huge mistake to not let him through. He’s got fan favorite written all over him, and like I said, I just love his song choices. Originality oozes out of him. He just needs a stage and he can perform, no doubt. Mad talented indeed, Randy. I was sad to hear his father passed away. I don’t remember that from his audition.
Heejun Han – And then there’s my true favorite. How can he not be everyones favorite? I’m not going to say that I think he’s winning this bad boy. But with every moment of him I see not singing, I like him more, and with every moment of him singing, I am more impressed by him. Unlike Creighton’s Billy Joel song, I thought Heejuns’ rendition was fantastic. I straight up love you, dude. I loved his optimism at the possiblity of not getting through
“What are you sweating?” “Mostly water”
“If they say no I will definitely hug Jennifer Lopez and kiss her because that’s every Asian mans dream, that’s everyone’s dream”
Sometimes I think he could be on a show. He could be on Community. He could be Ken Jeongs son. I loved that Steven recognized“You’re a better star than you are a singer” and my other favorite quote of his was “We’re all posers on the bus until we find a way to impress ourselves” I just love how happy everyone is for him, too. And I hope that douche Cowboy is crying as he sees Heejun succeed!
Phillip Phillips – I want to like him like I like Reed Grimm, but I just didn’t see him getting through to the big stage after some of his other performances. He’s talented instrumentally. I just don’t know. He doesn’t give me the same warm fuzzies that Reed did. On that same note though, I know he and Heejun are pals so I feel like he like I want to like him that much more. He also seems humble. And adorable.
Colton Dixon – And finally we had Mr. Dixon. I knew he’d be through before any performance. The judges owed it to him and his family to put that boy through. I will say playing the piano again, only made me like him more. That falsetto thing he does is great. He’s got a good “look into the camera” look too. Like he’s looking right at you. I wonder if they want him more because he didn’t really want them. I think they really regretted not having him go through the last time.
The boys we have left now, that I’m aware of, are below, and again I have bolded who I’d like to stay:
David Leathers, Jr.
Jermaine “Panty Melter” Jones
Kind of bad that I couldn’t even pick 6 more to stay. Maybe there are some I still haven’t seen? Only time will tell. And while I didn’t bold DeAndre, I’m next to positive that we’ll be seeing him on the big stage.


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Cambodia considered the fighting against money laundering and terrorism financing is the important, necessary and immediate task. The Royal Government of Cambodia showed goodwill and commitment to the fight against money laundering and terrorism financing through the Rectangular Strategies phase 4, the national strategies on the fight against money laundering and terrorism financing 2019-2023; the changes of legal framework and institutional framework for the effectiveness in terms of fighting the money laundering.
Reminding that Cambodia used to receive the assessment on the work of fighting against money laundering and terrorism financing for 2 times from APG in 2007 and 2017. Cambodia Financial Intelligence Unit (CAFIU) is the focal institution of Cambodia in facilitating the work on the anti-money laundering and terrorism financing. Anti-Corruption Unit is the law enforcement agency prepared its internal legal formalities to fight against money laundering and terrorism financing by arranging to have a parallel investigation, make changes to SOP in line with the standards of FATF, established the office against money laundering and created an inter-ministry technical working team to facilitate the law enforcement for investigating the offence of money laundering that the representative of ACU is the chief of the working group.
Relevant ministries/institutions of Cambodia closely cooperated in the work on fighting money laundering and terrorism financing and capacity building is also enhanced on a continuous basis such as simulations, trainings and workshops.
UN Representatives and governmental experts from Ewatini and Thailand in the process of reviewing Second Cycle of UNCAC Implementation in Cambodia.

domain: politics


text:
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domain: design


text: I’ve been listening to the Digital Story for years, but I don’t think I’ve ever reviewed it. I started my own podcast recently and quit after three episodes because podcasting is really hard and time consuming. It basically takes up an entire day. As a professional photographer, I just can’t give up an entire day each week. That’s why Derrick is amazing! I don’t know how he finds time to record a new episode each week with all his other responsibilities. My only complaint about the Digital Story has always been the length - I wish it was longer!!! Still do! But now I understand why it’s only 30 minutes! Because podcasting is serious work. The truth is, I don’t shoot like Derrick, I don’t use the same gear he does, I don’t even agree with his nimble philosophy. But I still tune in to every episode because I just genuinely like the guy. Unlike most photographers, he’s not egotistical, he doesn’t talk about how grand he is. He’s just a genuinely nice guy who happens to be a great photographer! I look forward to new episodes for years to come!!

domain: entertainment


text:
More to explore:
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Sheldons face. -- Even better when he tried to find plot holes in pride and prejudice and failed!! Because Jane is perfect :) roflburger.com
I think I actually teared up during this episode. "Please don't hurt my friend." Sheldon feels!!!!
big bang theory, alcohol always works.... at least temporarily anyway
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the big bang theory... oh raj, always inappropriate
Funny Pictures – 38 Pics yes please
Leonard and Bernadette are babies to go on it's a small world. I'm 11 and they're like 30 and yet I outgrew it before them!
The Big Bang Theory
Big Bang Theory LOL
omg! i crack up every time is see this! Amy Farrah Fowler is so hilarious! especially in this scene! it is awesome :p


text:
And much time has passed since I wrote here something. It is a workflow thing and I needed to reinstall the OS on my laptop which I used for writing here. Excuses and all but in any case, that is it.
Last summer I did an experiment: can I design logos in a very short time
Yes I can. The first took me about 30 minutes from reading the brief to saving the appropriate file for uploading to 99design.com. The last took me a little under 7 minutes. Now do not get me wrong, the inspiration came from somewhere else I am sure, from a “dimension” where time and space do not matter. I write this half jokingly but the serious part is about how do we get the ideas and do we need to measure a good idea and execution by the time it took to accomplish?
The short time I spent on it also reflects a half-life-long set of experiences, working effectively in Illustrator, reading carefully a brief and picking up wishes that the author either consciously knows about, or not. I can of course imagine them and that is fine too. It is not about Truth but about being inspired and being able to communicate.
So summer has ended and autumn has begun. My past days have been filled with learning to program at
Then I did a really great workshop in Illuka, Estonia about which I am going to write. That is my future as well. More workshops, more teaching. All about unlocking one’s own creativity and hopefully those at the workshops can pas the torch along and help others to do the same.
On the short list to-publish
- graphic design sprints I did which are hidden at 99designs.com
- writing about the workshops
- writing about my coding adventures (and perhaps my app idea)
I stopped doing the 99design.com projects are too many people wanted NDAs to be signed but it was never clear when I and the other designers could publish the results.

domain: artistic


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Surfers Against Sewage
"We are ocean activists. We are the voice of the ocean. Campaigning for a thriving ocean, thriving people.
Water Quality - we campaign for a clean ocean, rivers and lakes for wildlife and people. We demand an end to sewage discharge into UK bathing waters by 2030.
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According to a recent survey, corporate tax evasion is particularly difficult for Germany. In total, the tax authorities lose more than 21 21 billion a year because international corporations divert profits made in this country abroad and thus save on taxes.
Economists Gabriel Suckman, Thomas Torslow and Ludwig Wire account for 29 percent of total German corporate tax revenue. “Missing Profits” Research Project Has been investigated. In doing so, they evaluate data from statistical authorities. The figures indicate the year 2018. Hungary alone loses a large share of tax revenue at 31 percent.
The data also shows where corporate profits from Germany are taxed instead – at a much lower tax rate. Especially the tax centers of the European Union. According to researchers, Luxembourg ended with more than six billion euros, the Netherlands 5.5 billion and Ireland 3.6 billion. As a result, these countries also collect taxes.
Great Britain, France and the USA are among the countries affected by the tax evasion. Beneficiaries include Belgium, Switzerland and Singapore. In total, companies transfer 40 percent of their global profits to other countries, with 70 770 billion in 2018 alone, scientists write.
Prone to fits of apathy. Unable to type with boxing gloves on. Internet advocate. Avid travel enthusiast. Entrepreneur. Music expert.


Entire US Senate to descend on White House for N. Korea briefing
The Trump administration has asked every member of the US Senate to meet at the White House on Wednesday for a briefing on North Korea, in a rare and unusual move, amid mounting tensions with Pyongyang.
All 100 senators have been asked to attend the Wednesday briefing, which will be conducted by Secretary of State Rex Tillerson, Secretary of Defense Jim Mattis, Director of National Intelligence Dan Coats, and Chairman of the Joint Chiefs of Staff General Joseph Dunford, White House spokesman Sean Spicer said on Monday.
Although administration officials often travel to Capitol Hill to address members of Congress on foreign policy matters, it is unusual for the entire Senate to go to the White House, and for all four of the aforementioned officials to be involved.
The briefing was originally scheduled for a secure room at the Capitol, but was changed after President Donald Trump suggested a shift to the White House, according to congressional aides cited by Reuters.
Aides have also suggested the briefing is being held at the White House to make clear to North Korea that Washington is serious about a shift in policy when it comes to Pyongyang.
Senators told Reuters they were happy to be hearing from the White House.
“It’s [the location] their choice,” said Senator Ben Cardin, the top Democrat on the Senate Foreign Relations Committee. “I hope that we hear their policy as to what their objectives are, and how we can accomplish that hopefully without dropping bombs.”
Republican Senator Lindsey Graham said he hopes the administration will provide reassuring words that Pyongyang will not be given a chance to further develop ballistic missiles.
“By 2020, if nothing changes inside of North Korea, they’ll have the technology, they’ll have a breakthrough, to develop an ICBM (intercontinental ballistic missile) that can reach America. I hope this president will say that’s a non-starter,” he said.
The briefing will take place at 3:00pm local time on Wednesday.
Meanwhile, congressional aides said they are also working with the White House to arrange a similar briefing for the House of Representatives.
Trump has taken a hardline stance against North Korea since taking office in January, repeatedly vowing to “solve the problem” of Pyongyang.
He told the UN Security Council on Monday that the “status quo” is not acceptable, and that the council must be ready to impose new sanctions on North Korea.
Trump also had a phone call with Chinese President Xi Jinping over the weekend. The US president has been urging Beijing to put more pressure on Pyongyang, as it is North Korea’s main economic lifeline.
North Korean state media has responded to Washington’s moves by stating that Pyongyang is ready to sink an aircraft carrier headed towards the Korean peninsula, claiming its deployment was a “foolish military provocative act.”
Two Japanese Navy destroyers joined the US carrier strike group in an apparent show of force as North Korea prepared to mark the 85th anniversary of the founding of its military.


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Yvonne Nelson, a Ghanaian actress and a mother of one, has disclosed her intentions to have more children.
The Ghallywood actress, Yvonne, in an interview with Kwaku Sintim Misa on the KSM Show, said she was close to welcoming her baby number two last year, but work got in the way of her dream.
According to the 36-year-old, she wouldn’t like to have just a child but children, especially when she now has enough money to take care of five to six children.
“I want to have more kids. Last year, I was close…I have been ready, but last year was like, go for it, but I guess I was too busy.
“I wish I had four children because I’m working so hard. I have all these, whom am I going to leave it for? All these legacies I’m building can’t be for one child, so I don’t have to think like that. I want to have more like five or six. I Know am too old now. I wish I had more,” she said during the interview.

domain: celebrity


text:
(Posted 6th March 2018) Gillingham Town Youth FC are looking to boost their current under 7,8,9 and 10's squads for next season and will also be looking for players for a new under 7's (currently under 6's) team starting in the summer.
If you are interested please contact Chris Lee (GTYFC Head of Development) on firstname.lastname@example.org
(Posted 13th Feb 2018) Longfleet Youth FC are seeking female players for this seasons under 13 age group (School year 7 and 8), to train with an existing team, ready for next season. Winter training is currently at Carter Community School (3G pitch) on Tuesday evenings from 7.00pm - 8.00pm with summer training due to commence from Tuesday 6th March 2018 at Branksome Recreation Ground from 6.45pm - 8.00pm.
The team will be entering tournaments and playing friendly matches in order to prepare for the new season.
If you are interested, please contact the Club Secretary Dee Cooper at email@example.com or call on 07536 071320.

domain: sports


text:
This marks the 28th location for the company. With offices from coast to coast and work experience in over 40 states, Rapid Recovery said it continues to expand to meet the need for EPA-compliant refrigerant recovery. The company noted that its use of AHRI-certified gas-powered recovery equipment gives it a distinct speed advantage over typical recovery units.
“Industry leading EPA documentation and superior service are just two of the things that set Rapid Recovery apart,” said Adam Dykstra, president of Rapid Recovery. The company follows a franchise model, which Dykstra said supplies “great service backed by local ownership that provides a huge plus for local contractors.”
Dykstra added, “We are a genuine service company to HVAC, mechanical, and demolition contractors as well as in-house facilities departments and government agencies. As the burdensome cost of technicians’ time, equipment, maintenance, and expensive, yet inadequate off-the-shelf recovery equipment are realized, Rapid Recovery is in prime position for continued growth. Contractors are realizing that outsourcing the refrigerant recovery process to an expert can add profitability to their jobs, free up important technician time, and meet all of their documentation needs as well.”
Rapid Recovery’s Jacksonville, Fla., operation will be owned and operated by Hank Taylor and Chan Ritchie. “I like to say that we are really in the man-hours-savings business,” said Ritchie. “We can recover as much as 500 pounds of refrigerant in an hour, 10 times faster than your average recovery machine. This saves contractors a ton of costly time that can be repurposed to more productive activities than waiting around on recoveries... Then, add on top that we assume all of their EPA liability for every pound of refrigerant we recover and you see why we are expanding so rapidly.”
For more information, visit www.raprec.com.
Publication date: 10/24/2011
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domain: economics


text:
“The draft statement for the establishment of a nation by the Provisional Government of the Republic of Korea,” which is considered the foundation of the country’s first 1948 Constitution, has been designated as a cultural heritage.
"The draft statement for the establishment of a nation by the Provisional Government of the Republic of Korea” handwritten by independence activist Cho So-ang (whose real name is Cho Yong-eun) in Korean and Chinese based on his principle of three equalities will be registered as a cultural heritage,” said the Cultural Heritage Administration Tuesday.
The principle of three equalities, advocated by the theorist and political ideologue of the provisional government, refers to the equality of political, economic, and educational rights to realize an ideal society. The draft drawn by Cho is 10 squared manuscript papers long, which are 39.6 centimeters wide and 27.1 centimeters long, and consists of three chapters including the overview, restoration, and establishment of a nation. The statement was partly modified and passed at a meeting of the provisional government on Nov. 28, 1941.
“The document is of great value as it shows which kind of a nation the provisional government wanted to build and contains direct marks Cho So-ang made while revising the draft,” the Cultural Heritage Administration said.
Won-Mo Yu firstname.lastname@example.org

domain: history


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Furious Brexiteers have vowed the UK will derail European projects if Brexit does not happen, called for the removal of Theresa May and warned that the EU “will be facing perfidious Albion on speed”.
In a series of speeches at a meeting of the anti-Europe Bruges Group in Westminster there was also a warning that Europe was turning Britain from a Michelin-starred restaurant to one relying on microwave meals and there were cries of “traitor” at the mention of the Prime Minister.
Beginning the speeches on Tuesday, Mark Francois, the vice-chairman of the European Research Group (ERG) of pro-Brexit MPs, said people in the UK could not be “held captive against their will”.
— Jennifer McKiernan (@_JennyMcKiernan) April 9, 2019
Anger boiled over, with audience members shouting “f*** government” and repeatedly yelling “traitor!” at the mention of Theresa May.
Speaking from a podium with a photo of former prime minister Margaret Thatcher on the front, Mr Francois said he wanted EU leaders to know MPs like him would never vote for the Government’s Withdrawal Agreement so there was “no point” to an extension.
He said: “If however, you attempt to hold us in the EU against the democratically expressed will of the British people then in return we will become a Trojan horse within the EU, which will utterly derail all your attempts to pursue a more federal project.
“A new Conservative government led by someone like Boris or Dominic Raab might well vote down your projects, veto your attempts at greater military integration and generally make it impossible for you to bring about the more federal project in which you so desperately believe.”
Mr Francois then made a classical reference to treacherous diplomacy, to laughter in the room.
He said: “If you now try to hold us in against our will you will be facing perfidious Albion on speed.
“It would therefore be much better for all our sakes if we were to pursue our separate destinies in a spirit of mutual respect… Let my people go.”
Tory colleague Andrew Bridgen said the UK’s EU membership had turned the country from a Michelin-starred restaurant to one reliant on microwave meals.
He said: “We used to create these fantastic dishes from scratch and over the years this has been corrupted and we have been de-skilled.
“Now we accept our laws pre-packed from Brussels, ready to go in the microwave.
“We’ve become a chicken ding Parliament with chicken ding politicians.”
Mr Bridgen called for the removal of Theresa May as Prime Minister, to loud cheers in the room.
He said: “We need to remove Theresa May as soon as possible – she has already done enough damage.
“She needs to be replaced by someone who believes in Brexit and wants to deliver Brexit… I think we will probably lose some colleagues… and our majority.
“We will never get Brexit through that Parliament with those MPs so we’re going to have to go back to the people, it will be a People’s Vote but it wil be a general election.”
Conservative MP Anne Marie Morris described Brexit as “a complicated jigsaw puzzle” that must be played “in a very complex game of chess” and the only way to solve the problem was with a new party leader.
She said: “They (EU leaders) are as stubborn as Mrs May and I think they will keep extending, extending, extending.
“So I believe the only way out is frankly going to be with a new Prime Minister and we will go out on World Trade Organisation terms if we by then do not have a Parliament who will take us out with a proper trade deal.”
Ms Morris added she did not think Labour would bring a vote of no confidence in the Government and if they did she would not support it.
She said: “The truth is actually Mr Corbyn is a Leaver… and he does not really want an election.”


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Addley is a Liverpool based artist combining alternative songwriting and instrumentation with electronic elements and dance beats. Already so early on in his career, his heartfelt lyrics and huge instrumentation have been featured on BBC Introducing, with his upcoming single "Where You Were".
Addley's combination of acoustic and electric instruments in "Where You Were" creates a unique sound that makes you want to get up and dance, whilst also maintaining an emotional connection to his lyrics. "Where You Were" takes the intense emotions of grief and loss and transforms them into something accessible, somehow allowing you to feel without stripping away the fun of the song's instrumentation.
When asked how he would describe his sound, Addley told us that he wanted to experiment with balancing these two types of instrumentation on each of his tracks, saying,
"It can be really powerful to hear how synths, electronic beats and pads work with the more traditional band set up of piano, guitars, drums and live strings. I feel that with every track I'm working on I'm always conscious of finding out ways to work out the best and most interesting interactions of these different sounds".
We have been lucky enough to hear "Where You Were" ahead of its release, and agree that the track showcases this combination beautifully. When I first listened, I was immediately reminded of Florence and the Machine, who Addley tells us is one of his main influences.
"I think the main artists who influence my song writing are the likes of Florence and The Machine, London Grammar, Daughter… all the bands I’ve listened to on a constant loop for as long as I’ve really loved music. The past few years though I’ve become obsessed with electronic music and how artists like Bicep, Faithless and Disclosure can create these massive sounds and tracks that are so immersive and energetic. I feel like the energy and danceability of these artists is definitely something that is more often finding its way into my music, and I love it! Overall, it’s big alternative/indie bands but add some synths and beats to make it a bit more on the ravey side."
With a song that features such grand instrumentation and so many layers, we always love to know how these types of tracks sound in their beginning stages. Addley has walked us through his songwriting process.
"Most often my songs seem to start with a little hook I’ll get in my head, so for ‘Where
You Were’ it was just the main chorus melody of “it’s in my chest, round my neck and
the cold takes hold”. I’ll get a little melody line and then run to the piano and work out
how to play this with whatever chords go from there really. All the good ones seem to
go like that! After I’ve got the song finished just on the piano, just melody and chords,
I’ll record it and start to add and write the different parts until I’ve got a full track! The
song will normally take a few hours to write in its most basic form but then weeks
and weeks to finish adding all the other elements, which I think is the same for most
With huge names like Florence and the Machine and Faithless listed as his musical inspirations, we were interested to know what else made Addley start creating music.
"I’ve always loved music and wrote songs but it’s honestly the past few years I’ve
started to believe in myself that I could actually do it on my own. I think since I’ve
started to find more self-confidence and belief that I’ve began to take things seriously
and put my everything into creating and sharing my music. Now I’m just so excited to
Luckily for us, Addley has found that confidence to not only release his first single, but to pave the way for lots of exciting projects. When we asked Addley what we could look forward to from him, he told us,
"There’s a lot coming up I’m so excited about, I’ve gathered a big five-piece band
together and we’re getting ready to start live performances and recording live
sessions very soon! The next few singles are also done and nearly ready so you can
expect them very very soon. This is honestly just the start of everything I’ve got
planned and I’m so excited to share it!"
You can hear "Where You Were" from 27th January. Addley is definitely an artist to watch out for in the future.
Connect with Addley:


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All The Good Things by Ruth Hammond – Reviewed by Chris Mann
singer and multi-instrumentalist Ruth Hammond gained a bachelor’s
degree in Jazz Studies at Leeds College of Music.
On leaving college her saxophone talents were recognised by two
prizes “Top Saxophonist” and “Most Outstanding Performer”.
was runner up in the “Young Jazz Musician of the Year” awards in
has been a member of the hugely popular National Youth Jazz Orchestra (NYJO)
in the UK and has had a weeklong spot at the legendary Ronnie Scott’s
work can be heard soon on Morcheeba’s 2002 album.
Her own debut album “All the Good Things” is in my CD player
has a very
measured, bluesy sound which features Ruth Hammond’s lazy keyboards
and even lazier vocals. Her
delivery is smoky and the electronic effect applied during the chorus
sections can’t take away from that.
70’s rhythm guitar and in-your-face funky bass of Itchy Feet
made me smile. That busy
backing and the nonchalant vocal delivery made me think of Swing Out
Sister. The insane electric
piano, however, is totally original.
Your Love is
more mellow and thoughtful and the vocal on this song is a joy. It’s hard not to think of classic singer/songwriters such
as Carole King, Joni Mitchell and Judie Tzuke.
Superb backing vocals too. Pop
with a jazz tinge has worked for years – and never better than here.
takes an old skool twisty bassline and Brass Construction rhythm guitar,
slows it all down and preaches a very thought-provoking sermon while her
backing singers get all “gospel”.
Listen to the lyric on Another Day Gone – strong message
– strong song.
more likely to remember the electric piano than the vocal on More
than Sport – it’s well up in the mix and the riff is very
repetitive. In the chorus
the energy goes up a notch and this song gets better with repeated
Condition is a
very soulful tune, with a slow, strutting rhythm and that smoky Hammond
organ sound. The backing vocals are very strong but the message in
Ruth’s lyrics comes through unhindered.
By now, you’ll have realised what a versatile drummer Scott
a sweet fade into St Clears.
Stop thinking about categories and just enjoy a strong original
ballad, well written, tastefully produced and performed with feeling.
The sax solo (from Ruth herself) is enough and no more – the
song’s the star though. I
think this is my favourite.
getting into this offhand vocal delivery.
Don’t know why but Steely Dan popped into my head listening to Only
So Many Ways. They are
one of the bands Ruth Hammond was inspired by.
The organ is more to the fore – and I like it.
It’s one of the trendiest instruments around.
Er, where’d that sitar come from?
title track is whispered more than sung and it’s a very dreamy piece.
The vocal and gauzy Rhodes are placed in a big, airy soundscape
and the light cymbal work adds to the trance-like quality of the song.
I liked the inspirational words “your dark moments are not how
you’re defined…”. It’s
best to listen to this song on your own, with some tissues to hand.
It’s emotional stuff.
hard to know how to sum up. I’ll
start by saying I really enjoy listening to this CD – present tense, I
didn’t just listen to it once. That’s
the message really: this CD contains more than you think when you first
hear it – more lyrical depth, more musical confidence and more
original talent, despite the fact that certain influences are apparent.
If you have a friend who owns this CD, make sure you go and see
them soon – and take a good bottle of wine…
Records THRCD1 – no producer credits
available from www.ruthhammond.com


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What is Gynecomastia?
Gynecomastia is a condition characterized by excessive male breast tissue.
Gynecomastia is triggered by a decrease in the amount of the hormone testosterone compared with estrogen. The cause of this decrease can be conditions that block the effects of or reduce testosterone or a condition that increases your estrogen level. Dr. Prendergast recognises the sensitive nature of this issue and offers patients the best way to get rid of gynecomastia.
Why consider Male Breast Reduction?
If you are a man who is overly-self conscious and limited in your life by excess breast tissue, consider VASER.
Estrogen controls female traits, including the growth of breasts.
Most people think of estrogen as an exclusively female hormone, but men also produce it though normally in small quantities. However, male estrogen levels that are too high or are out of balance with testosterone levels can cause gynecomastia. Some men in Ireland spend quite some time puzzled by this ‘chest fat’, unaware there is an endocrine cause and fast way to remove these ‘moobs’. There is no exercise or workout that can reduce ‘man boobs’ – removal of fat cells is the only permanent solution.
Hormones can be extremely difficult to manage as there are myriad factors which can disrupt the delicate balance. These range from genetics, to common medications, to even using too much lavender or tea-tree scented shower products. Men generally cannot obtain visible loss of excessive breast tissue, or Man Boobs, through holistic or medication-based treatment. Targeted liposuction with a unique capacity to breakdown fat is needed.
The emotional upset and embarrassment caused by gynecomastia warrants a fast, long-lasting treatment, and we provide that to our male patients at Venus Medical. VASER is considered the best method of male breast reduction as the ultrasound technology effectively breaks down the particularly dense adipose tissue.
How is the Male Breast Reduction performed?
As an Official VASER Instructor and first surgeon to offer it in Ireland, Dr. Peter Prendergast applies his expertise to quickly remedy gynecomastia. VASER involves no general anesthetic or IV sedation. Tumescent local anaesthetic allows for rapid treatment and extremely low risk and recovery time. Because nerves and blood vessels are largely undisturbed, patients and physicians report low to minimal pain and bruising. The VASER liposuction method allows Dr. Prendergast to contour and remove the fat with superior precision. VASER has already transformed the lives of countless men who have presented to our clinic suffering from male chest fat.
What is VASER Lipo?
VASER Lipo is a minimally invasive means of liposuction, performed without the use of sedation or general anesthetic. During the procedure you are awake and lucid, and recovery time is extremely quick. The results of VASER for male breast reduction are particularly impressive, since ultrasound efficiently and smoothly removes the dense tissue typical in the male breast area.
Why consider VASER Liposuction?
VASER Lipo is an advanced, highly efficient and specialized cosmetic procedure, of which Dr. Prendergast is an international pioneer. Compared to traditional liposuction, there is a quicker recovery, less swelling and bruising and it is associated with smoother results and superior skin tightening. The VASER method allows for more specific destruction of fat cells and limited injury to all other tissues. It also avoids many potential risks by using only local anaesthesia. This also reduces the higher costs associated with general anaesthesia.
Here are yet more reasons to consider VASER Lipo:
1.Dr Prendergast was the first doctor in Ireland to perform the VASER procedure
2.Dr Prendergast is the only official VASER instructor in Ireland
3.As President of the European College of Aesthetic Medicine and Surgery, Dr Prendergast trains surgeons in VASER internationally
4.Dr Prendergast has written several book chapters on the VASER technique
5.Dr Prendergast is the only doctor in Ireland trained in 4D Sculpt, the advanced VASER Hi-Def technique. He was invited onto the faculty of the renowned International Center for Artistic Body Sculpting in Colombia
6.Dr Prendergast uses tumescent anaesthesia for male breast reduction, the safest way to perform any liposculpture technique and the most cost efficient
7.We do not consider this merely fat removal, rather, it is an artistic form of true body sculpting
8.All consultations performed only by Dr Prendergast
9.Unsurpassed preoperative and postoperative care for best results and safety
10.We love VASER. We are sure you will, too!
How is the VASER Lipo performed?
1.Infiltration: slowly passing the diluted anaesthetic solution into the fat through tiny incisions using special cannulas or sprinklers
2.Emulsification: once the areas are completely numb, the fat is liquefied using special fine VASER probes. These probes emit ultrasound waves, efficiently breaking down the fat whilst preserving other tissues such as blood vessels and nerves. Contouring the body in this way requires an artful eye and delicate, graceful movements of the probe.
3.Aspiration: Once the fat has been emulsified, it can be easily extracted through gentle suction.
How long is the recovery period?
Most patients are up and around as soon as they get home, are able to leave home the next day, and return to work within 2-3 days. We will give you detailed instructions on when to begin activity and when you may return to work and will prescribe the necessary mild pain medication.
How much does it cost?
The costs of treatment will be outlined during a consultation with Dr Prendergast, following assessment.
How long do the results last?
VASER, like all other forms of liposuction, removes the male chest fat and breast tissue permanently. There will be a small amount of fat deliberately left behind under the skin to ensure a smooth, even result.
Call our clinic advisor on 012962747 to book a consultation.

domain: medical


text:
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Periaxin is expressed in mammalian Schwann cells and lens fiber cells, and has been identified in a screen for cytoskeleton-associated proteins. Charcot–Marie–Tooth 4F is caused by losses or mutations of the periaxin gene. The periaxin gene encodes two protein isoforms, namely, L-periaxin and S-periaxin. S-periaxin contains 147 amino acid residues and has an N-terminal PDZ domain. In this paper, S-periaxin was reported to be homodimerized through the formation of intermolecular disulfide bonds with its Cys88 and Cys139 residues under mild oxidation conditions. The covalent dimer of S-periaxin was also observed by western blot analysis and bimolecular fluorescence complementation analyses. S-periaxin dimerization formation could be regulated by cellular redox fluctuations. These results offer a possible mechanism to the formation of periaxin complexes, improvement of complex stability, and establishment of a link between the extracellular matrix and the cytoskeleton.
Periaxin was initially identified from myelinating Schwann cells in the mammal peripheral nervous system (PNS), and it plays an essential role in stabilizing the Schwann cell–axon unit through membrane–protein interactions [1,2]. Periaxin protein constitutes 16% of the protein content of the PNS . Periaxin gene deletions or mutations cause demyelinating peripheral neuropathies, such as Charcot–Marie–Tooth disease 4F, peroneal muscular atrophy, hereditary motor and sensory neuropathy [1,4], and recessive Dejerine–Sottas neuropathy . In lens fiber cells, periaxin interacts with ezrin, periplakin, and desmoyokin, which constitutes a macromolecular cytoskeletal complex of ezrin–periplakin–periaxin–desmoyokin (EPPD) . The effect of assembling is found to be involved in the maturation, packing, and membrane organization of lens fiber cells .
Periaxin has two isoforms: L-periaxin (147 kD and 1461 amino acid residues) and S-periaxin (16 kD and 147 amino acid residues), which is a truncated isoform . Both proteins have an N-terminal PDZ (PSD-95/Discs Large/ZO-1) protein-binding domain (Fig. 1A) [7,8]. Aside from the PDZ domain, L-periaxin can be further divided into a basic nuclear localization signal region, a long-repeat domain, and a C-terminal acidic domain [9,10]. It has been found that periaxin can dimerize with its PDZ domain . In the absence of the PDZ domain, ΔPDZ-Prx mice are barely functional . This phenomenon may be attributed to the distinctive clustering of the periaxin complex in the Schwann cell plasma membrane [7,11,13].
The PDZ domain consists of an ~90-amino acid protein-binding motif that interacts with the cytoplasmic tail of plasma membrane proteins or with the cortical cytoskeleton, which is involved in the assembly of macromolecular signaling complexes [8,14]. The PDZ domain of periaxin is poorly conserved, and the highest sequence identity is found in the N-terminal PDZ-like domain of giant AHNAK nucleoprotein 2 (Fig. 1A) . The PDZ domain of periaxin or AHNAK includes a unique subfamily that may link the extracellular matrix to the cytoskeleton network . The dimerization structures of periaxin–PDZ domain have been presented (PDB: 4CMV), showing that an intertwined, domain-swapped dimer exhibits a head-to-tail antiparallel orientation (Fig. 1B) .
The function of the PDZ domain includes the recognition of internal peptide motifs, hetero- and/or homodimerization, as well as the interactions with membrane phospholipids [16,17]. The PDZ domain of periaxin also guides its translocation from the nucleus to the cytoplasm . A nuclear export signal (NES) is identified specifically from amino acid 73–86 of the L-periaxin–PDZ domain, and the nuclear export activity of L-periaxin is found to be inhibited by NES mutation or by LMB treatment . L-periaxin and S-periaxin have the same PDZ domain (Fig. 1A), but S-periaxin is restricted only to the cytoplasm or nucleus of Schwann cells. This restriction can be attributed to protein domain simplification and functional uniqueness , and the function of S-periaxin is likely to regulate an mRNA splicing .
To date, most functions of S-periaxin are unknown. However, S-periaxin is likely to participate in large molecular complexes through the interactions of its PDZ domain, which is directly involved in biological functions . In the present study, we found that S-periaxin can form dimers or oligomers under non-reducing conditions, and cysteine residues 88 and 139 may be involved in the intermolecular disulfide bond formation in vivo and in vitro.
The DNA sequence encoding S-periaxin was amplified by PCR by using the following primers: forward primer (5′GAGGAATTCATGGAGGCCAGGAGCCGGAGTGCCGAGGAGCTGAGGCGGGCGGAG3′) and reverse primer (5′ CGAGTCGACTCACGGCGCAGAGACCGGATCGCTGGGGCAGTCCAGGGC3′). The template is a cDNA library from rat RSC96 Schwann cells (Type Culture Collection of the Chinese Academy of Sciences, Shanghai, China). The resultant PCR product digested with EcoRI and SalI (Thermo Scientific, Waltham, USA) was inserted into vector pET-M-3C, in which thrombin protease cleavage sites replaced by human rhinovirus 3c protease cleavage sites,generated from pET15b (Novagen, Madison, USA) to obtain the recombinant plasmid pET-M-3C-S-periaxin. The 441 bp open reading frame was verified by DNA sequence analysis (HuaDa, Beijing, China). The mutants (C88/G, C97/G, C139/G, C88,97/G, C88,139/G, C97,139/G, and C88,97,139/G) were generated by Easy Mutagenesis System (TransGen Biotech, Beijing, China). Furthermore, S-periaxin or its mutants were also cloned into mammalian pCMV-Tag-3B (Myc tag) and pEGFP (EGFP tag) vector (Invitrogen).
The production of recombinant S-periaxin or its mutants was expressed in Escherichia coli BL21 with 0.3 mM IPTG. The cells were harvested and resuspended in lysis buffer (20 mM Tris–HCl, 500 mM NaCl, 5 mM imidazole, pH 7.9). The constructs were lysed by sonication, and the debris was removed by centrifugation at 17,000 g at 4°C for 30 min. The supernatant was applied to a nickel-nitrilotriacetic acid (Ni-NTA) affinity column (GE Healthcare, Bethesda, USA). Recombinant protein was eluted with elution buffer (20 mM Tris–HCl, 500 mM NaCl, 500 mM imidazole, pH 7.9). After concentration by ultrafiltration, the protein was loaded onto Sephacryl S-200 gel filtration chromatography with a buffer containing 20 mM Tris–HCl (pH 8.0), 100 mM NaCl, 1 mM ethylenediaminetetraacetic acid (EDTA), 2 mM DTT, and 15 mM β-mercaptoethanol. The purified protein was collected, concentrated, and then stored at 4°C.
HeLa cells were maintained at 37°C with 5% CO2 in RPMI-1640 containing 10% fetal bovine serum. Cells were seeded onto six-well plates and cultured overnight prior to transfections. At 60%–70% confluence, the cells were transfected individually with Myc-tagged wild-type (WT) S-periaxin or its mutants by using TurboFect transfection reagent (Thermo Scientific) according to the manufacturer's instructions. Approximately 48 h post-transfection, HeLa cells were harvested, and the free sulfhydryl groups of proteins were blocked by incubation with 30 mM N-ethylmaleimide (NEM) on ice for 30 min . Then cells were lysed in 400 μl of lysis buffer (20 mM Tris–HCl, 150 mM NaCl, 1% Triton X-100, 1 mM PMSF, sodium pyrophosphate, β-glycerophosphate, EDTA, and leupeptin). The supernatant of cell lysates was separated by 12% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) without a reducing agent. S-periaxin was transferred onto a PVDF membrane (Millipore, Billerica, USA). The membrane was incubated with rabbit anti-Myc antibody (TransGen Biotech) as the primary antibody, followed by incubation with horseradish peroxidase (HRP)-labeled secondary antibody (TransGen Biotech). Finally, the immunoblots were visualized using an enhanced chemiluminescence reaction kit (Engreen Biosystem, Beijing, China).
HeLa cells were transiently transfected with pCMV-tag3B-S-periaxin/pEGFP- S-periaxin or pCMV-tag3B-S-periaxin/pEGFP. At 48 h post-transfection, whole cell lysates were harvested in 400 μl of lysis buffer, kept on ice for 15 min and centrifuged at 4°C. The supernatant was collected and incubated with 2 μl of anti-Myc antibodies or 2 μl of mouse IgG as the control and allowed to rotate overnight at 4°C. Protein A+G agarose (Beyotime, Shanghai, China) was added and incubated for 4 h, followed by washing with lysis buffer. The samples were separated by 12% SDS-PAGE and analyzed by western blot analysis with the corresponding antibodies. The antibodies used were rabbit anti-Myc antibody and mouse anti-EGFP antibody (TransGen Biotech).
Bimolecular fluorescence complementation (BiFC) plasmids are shown in Fig. 2. The mCherry proteins were split at positions between amino acids 159 and 160 . The N- and C-terminal coding regions of mCherry were amplified by PCR from plasmid pmCherry-C1 (Invitrogen). The gene fragment of mCherry1-159 was cut by SalI and HindIII (Thermo Scientific) and then inserted into the same pQE-30 sites to generate pCherry1-159. The gene fragment of mCherry160-237 was digested by BamHI and EcoRI (Thermo Scientific), and then inserted into the corresponding sites of pET-28a to generate pCherry160-237. The DNA sequence of S-periaxin (WT and its mutants) was inserted into pCherry1-159 or pCherry160-237 to construct pCherry1-159-S-periaxin (WT and its mutants) and pCherry160-237-S-periaxin (WT and its mutants). All sequences were verified by DNA sequencing.
Escherichia coli cultures that express various fusion proteins of pCherry1-159 or pCherry160-237 (Fig. 2) were maintained on Luria-Bertani medium at 37°C, induced with 0.3 mM IPTG, and grown for an additional 3 h. The bacterial cells were collected and resuspended in 100 µl of phosphate saline buffer . The fluorescence of the E. coli cells was visualized using the Delta Vision Microscopy Imaging Systems (GE Healthcare) with the TRITC channel.
Periaxin isoform 1 and 2 (NP_066007.1 and NP_870998.2) were identified from Homo sapiens through an NCBI database search. The homology of periaxin to other proteins is very low, and the only conserved domain is the PDZ domain of N-terminal . Sequence alignment revealed an overall sequence identity of 30% between S-periaxin (NP_066007.1) and AHNAK isoform 2 (NP_076965.2) at the amino acid level (Fig. 1A). Furthermore, the crystal structures of the PDZ homology domain of periaxin (PDB: 4CMV; 14–104 aa) were verified . We paid special attention to the structure model. As shown in Fig. 1B, the core of the PDZ dimer is formed by a 6-stranded antiparallel β sheet by chain A and B, and the cores of α2 helices (Phe75–Tyr90) from the two chains of PDZ dimer structure are placed side by side . An extended conformation in the N-terminal (Met1–Arg13) and C-terminal (Gly105–Pro147) of S-periaxin is modeled based on SAXS data , thereby suggesting that S-periaxin is elongated and the moiety points of carboxyl terminus remain exposure the most.
L-periaxin dimerization had been shown by yeast two-hybrid experiments, pull-downs from nerve lysates, and CO-IP from mammalian cells that are over-expressing L-periaxin constructs . To confirm the dimerization form of S-periaxin, HeLa cells were transiently transfected with pCMV-tag3B-S-periaxin/pEGFP-S-periaxin. Myc-tagged S-periaxin was shown to co-precipitate with EGFP-tagged S-periaxin. No specific binding was observed with the EGFP control or the irrelevant mouse IgG. These results indicate that S-periaxin forms homodimers in vivo (Fig. 1C).
The recombinant His-S-periaxin was expressed in BL21 cells and its molecular weight was estimated to be ~15 kD (Fig. 3A). Without any reducing reagent in the loading buffer, different oligomers of S-periaxin were detected, which suggests that S-periaxin may oligomerize under oxidation (Fig. 3A, lanes 2–5). Approximately 10 mM DTT was added to Laemmli loading sample buffer to reduce the tetrameric and trimeric protein bands. A few S-periaxin dimers were detected after incubation for at least 5 min (Fig. 3A, lanes 6–9).
The recombinant S-periaxin in 20 mM Tris–HCl (pH 8.0), 100 mM NaCl, 2 mM DTT, and 15 mM β-mercaptoethanol was desalted by using a P10 column and quickly transferred to a buffer that contains 20 mM Tris–HCl (pH 8.0) and 100 mM NaCl. Then it was treated with different concentrations of H2O2 (0–1 mM) for 30 min at room temperature and analyzed by non-reducing SDS-PAGE. H2O2 treatment induced the formation of an S-periaxin dimer in a dose-dependent manner (Fig. 3B). The S-periaxin dimer in 20 mM Tris–HCl (pH 8.0) and 100 mM NaCl can be fully converted back to the monomer with the addition of 5 mM DTT (Fig. 3C). These findings indicate that the intermolecular disulfide bond formed by oxidation is involved in the oligomerization of S-periaxin.
To investigate the role of the three Cys (C88, C97, and C139) in the formation of intermolecular disulfide bond, S-periaxin and its mutants (C88/G, C97/G, C139/G C88,97/G, C88,139/G, C97,139/G, and C88,97,139/G) were subject to oxidation with H2O2 in vitro. In single mutations, only Cys88 and Cys139 showed an obvious weak tendency to form an H2O2-induced dimer, unlike WT (S-periaxin-Cherry1-159/Cherry160-237-S-periaxin) and C97/G (Fig. 4A–C). Mutation of C88,139/G in S-periaxin also had a greater tendency to reduce dimer formation than C88,97/G, and C97,139/G (Fig. 4D–F). When all Cys were mutated to Gly, C88,97,139/G only showed a monomer band (Fig. 4G). These results indicate that Cys of S-periaxin are involved in dimerization through the formation of intermolecular disulfide bonds. Cys88 and Cys139 undergo oxidation in S-periaxin, while Cys97 was relatively inert to oxidation.
To determine whether S-periaxin can form a dimer in vivo, Myc-tagged S-periaxin (WT, C88,97/G, C88,139/G, C97,139/G, or C88,97,139/G) was transfected into HeLa cells. Before lysis, these cells were collected and incubated with 30 mM NEM to block reactive sulfhydryl groups . The cell lysates were loaded onto non-reducing SDS-PAGE. The dimer of S-periaxin and its mutants was detected by western blot analysis using anti-Myc antibody. Interestingly, on non-reducing SDS-PAGE, both monomer and dimer were detected in WT Myc-S-periaxin and in three mutants (C88,97/G, C88,139/G, and C97,139/G), while only monomer was detected in the C88,97,139/G mutant (Fig. 5), suggesting that Cys residues are involved in the assembling of S-periaxin. However, the intermolecular disulfide bond is not the only important factor involved in the oligomerization of the molecule.
Subsequently, BiFC analysis was performed to obtain the intermolecular disulfide bond information of S-periaxin in E. coli BL21 cells. S-periaxin and its mutant fusion with mCherry fragments were transformed into BL21, and all the fusion proteins were expressed in supernatant fluid (Fig. 6C). The emission of mCherry-RFP fluorescence by the various strains was monitored [22,23]. Co-inoculation of pCherry-1-159-S-periaxin/pCherry-160-237-S-periaxin constructs resulted in strong red fluorescent signals located in small aggregations all over the cytoplasm, thereby indicating the self-interaction of S-periaxin. S-periaxin can form dimers in living cells (Fig. 6A). Red fluorescence was found in little aggregates from Cys mutation C97/G, C88,97/G and C97,139/G. However, the oligomerization of C88/G, C139/G, and C88,97,139/G was relatively lower than that of S-periaxin (Fig. 6B). In contrast, the mutations of S-periaxin did not completely abolish BiFC fluorescence, which means that other factors may also contribute to S-periaxin dimerization, among which the spatial structure of the PDZ domain is one such factor .
L-periaxin and S-periaxin have the same N-terminal PDZ domain. However, L-periaxin predominantly localizes in the nucleus (adaxonal membrane) during axon ensheathment and early myelination, and then localizes to the abaxonal Schwann cell membrane (opposite the basal lamina) that is undergoing myelination . This process is unlikely for S-periaxin, which distributes in the cytoplasm (occasionally in the nucleus) .
The PDZ domain is a protein–protein interacting module that plays an important role in the organization of signaling complexes . For example, PICK1 (protein interacting with C-kinase 1) PDZ was reported to be redox-regulated by the formation of a reversible intermolecular disulfide bond that engages Cys44 in the β2–β3 loop under mild oxidative conditions . The most common target for cellular oxidants is Cys residues in proteins. Upon exposure to oxidants such as H2O2, free sulfhydryl groups in the Cys residues can be oxidized into sulfenic, sulfinic, or sulfonic acids. H2O2 treatment induced the formation of intermolecular disulfide bonds between S-periaxin in vitro. In this report, we demonstrated that Cys88 and Cys139 in S-periaxin play a role in oxidation-induced disulfide bond formation, while Cys97 is relatively inert to oxidation.
Furthermore, from the PDZ crystal structure of periaxin (Fig. 1B), we found that the α2 helices (Phe75–Tyr90) from the two chains of the periaxin dimer structure are side by side, and the distance between Cys88A and Cys88B is ~5.03 Å, which meets the standard of a disulfide bond. In comparison, the distance between Cys97A and Cys97B or Cys88A–Cys97B is more than 10 Å, suggesting that disulfide bonds cannot be formed between these sites. The conformation of the C-terminal (including that of Cys139) exhibits extended structures. Although the homology of S-periaxin to other proteins is low, the C-terminal extensions gradually increase the possibility of oligomer formation through the Cys139–Cys139 intermolecular disulfide bond.
Various techniques, including yeast two-hybrid screening, Co-IP, glutathione S-transferase pull-down, and the recently developed tandem affinity purification assays, have been used to detect and study protein–protein interactions [11,25,26]. BiFC, which was developed by Hu et al. , detects weak or otherwise transient protein–protein interactions in vivo. BiFC is based on the principle that the N- and C-terminal fragments of fluorescent proteins and its derivatives alone do not fluoresce. However, if they are fused to interacting proteins, the two non-functional halves can be brought into close proximity and form a functional fluorophore as a result of specific protein interactions [28,29]. In this study, we performed BiFC analysis to obtain the intermolecular disulfide bond information of S-periaxin in bacterial cells. Results showed that S-periaxin (WT and its mutants) could form inter-subunit disulfide bonds in vivo. However, inter-disulfide bond formation was not the main reason for the dimerization or oligomerization of S-periaxin.
Other factors may also be involved in the oligomerization of S-periaxin in the cytoplasm. The mutation of the GLGF motif of the L-periaxin–PDZ domain into GLAA attenuated homodimerization (Fig. 1A), suggesting that the canonical peptide binding groove of the periaxin–PDZ domain may participate in homodimerization . To date, the function of S-periaxin has not yet been revealed. The dimerization of S-periaxin may provide a clue to the hypothesis that S-periaxin may interact with L-periaxin by PDZ dimerization to regulate the function of L-periaxin in Schwann cells . These questions are critical to future studies.
S-periaxin was found to be homodimerized through the formation of intermolecular disulfide bonds between its Cys88 and Cys139 residues under mild oxidative conditions. Cys97 residue is relatively inert to oxidation. We propose that the molecular conformation of S-periaxin could be regulated by cellular redox fluctuations under physiological and pathophysiological conditions.
This work was supported by a grant from the National Natural Science Foundation of China (No. 31170748).

domain: chemistry


Gophers Hang Another Banner
Detroit, Mich. -- Seth Ambroz, Travis Boyd, Christian Isackson, Ben Marshall, Kyle Rau, and Sam Warning accomplished something that no previous Gopher class had ever done by winning a fourth consecutive regular season title last weekend. This weekend they accomplished something they had never done.
Minnesota (23-12-3 overall) defeated Michigan (22-15-0) 4-2 at Joe Louis Arena in front of 6,355 at Joe Louis Arena to secure an NCAA Tournament bid as an auto qualifier and win their class’ first tournament title and the program’s first ever Big Ten tournament title.
“That’s always the goal when ...


text:
If you have ever been in West Africa, you know "wax" fabric well.
It's in every market and adorning many women and men,
particularly at special occasions.
"Wax" refers to the manufacturing process,
(not the texture of the fabric)
But, did you know...
The *Akan of Ghana and Côte d'Ivoire use cloth
not only for personal adornment ,
but also, metaphorically,
as a powerful expressive medium of communication... .
cloth one wears are likely to say something about one's identity,
beliefs, or set of values.
that elicits the spoken proverb.
either through written or representational imagery,
Akan woman's dress designers also shape the garment itself
to convey some messages.


text:
Just to let you know.... The hands are a great! I was so proud I showed them to everyone! "look at my Little monkey Hands!!" I was just silly!
KP does mean knit and purl into the same stitch.
I used red heart acrylic medium worsted since it was for my kid. I did not swatch so he is pretty big.
for the hands (since I used acrylic instead of cotton) I had to adjust the pattern.
(use the pattern for a guide)
I cast on 18 stitches
row 1-12 :stocking stitch
first finger:(imagine first finger as the pinkie)work 12, turn. (sl1, P5, turn) (sl1, K5, turn) for 10 rows. thread yarn through all finger stitches and leave a long enough end to sew.
second finger:pick up 3 stitches, turn. P3, P3 from other needle. Work the same way as first finger
third finger: just like second finger.
for the thumb I just knit a rectangle 6 stitches 10 rows and attached it to the side of the hand.
stitch all the sides of the fingers. Now you have a cute monkey hand to show all your friends!
as soon as I can figure out picture post (I am a much better knitter than photographer!) I will try to get some pictures up! Good luck! Feel free to ask if you need help! Gotta keep the crafster good karma flowing!
Here is a closeup of the hand
and here is monkey in all his naked glory!


text:
Today's exchange rates updated 20th Oct 2018 07:01 (1 British Pound = 1.13 Euro - 1 GBP = 1.13 EUR)
In Germany the currency is Euro (abbreviated as EUR). In UK the currency is British Pound (abbreviated as GBP). In order to facilitate the currency move, or send money from UK to Germany, a British Pound to Euro currency transfer is needed.
|Broker/ bank||Transfer Fees||Transfer Funds||Transfer Options||Quote/ Apply|
Over the Phone
Pound Sterling (GBP) The Pound has had a volatile week, as the latest UK data has given investors a mixed idea of whether a potential 2018 interest rate hike from the Bank of England (BoE) is...
Pound Sterling (GBP) Despite some disappointing UK retail sales results on Thursday following Wednesday’s surprising slowdown in UK inflation, the Pound was able to hold above its worst levels...

domain: finance


text:
A Tale of Two Furry Friends: The Dog and the Cat
Once upon a time, there was a dog and a cat who lived in the same house. They were different animals, of course, but they got along very well.
Every day, the dog and cat would go for long walks together in the park. They'd chase each other around trees, and bark and meow loudly for all to hear!
One day, when they were playing in the park, they stumbled upon a magical cave. The dog and cat were both curious about what was inside so they decided to explore.
Inside the cave they found a secret passage that led them to an underground world which was filled with all sorts of strange creatures!
The dog and the cat were amazed by all the sights they saw and couldn't help but explore further. As they walked deeper into this underground world, they noticed something strange—the creatures could talk!
The creatures welcomed the dog and cat to their world and showed them around all of their strange lands and kingdoms!
After exploring this magical place for what felt like hours, the dog and cat decided it was time to go home as it had already gotten dark outside.
On their way back through the secret passage, they said goodbye to all of the wonderful creatures that had welcomed them so warmly in their underground world!
From then on out, whenever the two friends went on their daily walks in the park, they would secretly hope to stumble upon that mysterious cave again—but it never did happen again… not until one day when…
…When one special night while playing in the park, the dog barked loud enough that it actually woke up some of those mysterious creatures from within their underground world! And just like that ,the adventure began again...


text:
one4two is a sound design studio located in Amsterdam. Their housestyle therefore takes the form of a soundscape using the classic Spirograph device to create colorful and symbolic designs in space, reminiscent of the designs sound makes.
KesselsKramer invites you to
Have a similar project, problem or communication-need laying about? Would you like us to help? Or, simply like the look of our work and want to know more, get in contact below.


Information provided by LSUsports.net
BATON ROUGE, La. - The No. 13-ranked LSU softball team run-ruled the No. 16-ranked Arkansas Razorbacks, 10-2 in five innings to claim the series on Saturday night in Tiger Park. With the win, LSU improves to 39-13 and 12-10 in conference play while Arkansas falls to 37-13 and 12-11 in the SEC.
Arkansas answered the very next inning with a lead-off solo home run and a sacrifice fly to tie the game up at two apiece.
In the bottom of the fifth inning, Shelbi Sunseri hit a two-run home run, scoring Shemiah Sanchez and extending the lead to 8-2. Senior Sydney Loupe would later come in to pinch hit and picked up her first career hit as a Tiger and was eventually the game-ending run thanks to a single by Griggs, giving the Tigers the 10-2 win.


text:
How great are bliss balls when you don’t have to get out the food processor for them? The last time I shared my recipes for No Blend Bliss Balls, they were a hit because they were so simple to whip up with minimal effort and cleaning up involved! Today’s little recipe is exactly the same. You really only need a bowl, a mixing spoon and a container to store the delicious little bites in.
I call these all natural protein balls, because I don’t actually use any protein powder in them. I’ve used a mix of wholefoods that are naturally high in protein, like LSA (ground linseed, sunflower seeds and almonds), sunflower seeds, chia seeds, hemp seeds and peanut butter (I alternate between Mayver’s and Macro peanut butter – both are delicious and don’t contain any sugar or additives).
To sweeten it, I added just a couple of spoonfuls of organic honey. Bliss balls generally have a base of medjool dates which are delicious but quite high in natural sugar, so if you’re looking for something less sweet, these are a great alternative!
As well as being filled with protein, they’re also packed with good fats, making them a good source of long sustaining energy and they’ll keep you fuller for longer.
To make them, I simply threw all the ingredients together in a bowl, mixed it and scooped out spoonfuls and rolled them into bite sized balls and popped them in the freezer to store until needed.
They taste like peanut butter cookie dough and I seriously can’t get enough of them! Please remind me again why I don’t make these more often?
Hope you had a good start to the week everyone!
- 1 and half cups LSA
- 1/4 cup chia seeds
- 1/4 cup hemp seeds
- 1/4 cup sunflower seeds
- 1 cup natural peanut butter
- 2 tablespoons honey*
- Place all ingredients in a large mixing bowl and stir until well combined.
- Using a spoon, scoop out mixture and roll/shape into a ball.
- Repeat until mixture is finished.
- Store in freezer until needed.
- * Make this recipe vegan by swapping the honey for rice malt syrup or blackstrap molasses.

domain: food


text:
CHANDIGARH: A controversial bill dealing with citizen and residency rights has triggered mass protests in northeastern India.
The Citizenship (Amendment) Bill 2016 proposed by the ruling Bharatiya Janata Party (BJP) was passed in the lower house of the Indian Parliament and will now be taken to the upper house.
The amendment relaxes eligibility rules for immigrants belonging to six minority (non-Muslim) religions from Afghanistan, Bangladesh or Pakistan.
Various parties in the region have launched a strong protest in the northeastern state of Assam, questioning the bill which grants citizenship on the basis of religion.
Activists and political parties say the bill violates the “secular constitution” of the country.
– Illegal immigrants
The bill will be applicable to the entire country but the northeast is protesting as a large number of Bengali Hindus have illegally entered the region, experts say.
All Assam Students Union (AASU), one of the protesting bodies, term the bill as “damaging and unethical”.
“The bill not only will allow a section of illegal foreigners to become Indian citizens… this will completely change the demographic pattern of the region, and the indigenous people will become minority in their own motherland,” Dipanka Kumar Nath, a student leader told Anadolu Agency.
“It is our firm stand, whoever it may be, either Muslim, or Hindu, illegal foreigners are illegal. A bill to legalize illegal foreigners is really unfortunate, and that too in consideration of religious identity is really unconstitutional and unethical,” said Nath.
The move by BJP to bring the bill has also angered its allies in the northeast.
The Asom Gana Parishad (AGP), a local party, recently broke alliance with the BJP saying the bill is unconstitutional. “You cannot define the citizen on the basis of religion. We have already resigned…if the government doesn’t listen to people, the situation will be more volatile,” AGP head Atul Bora told Anadolu Agency.
– Opposition cries foul
Rahul Gandhi, head of the main opposition party, Indian National Congress, said last week he was disturbed by the growing unrest.
“I am disturbed by the growing violence & unrest in the N East where religion & ethnicity are being used as weapons by the BJP to polarise the region for political gain. This type of politics, that pitches brother against brother & fans hatred, is evil & must be stopped,” he tweeted.
Maulana Maqsood ul Hasan Qasmi, head of India’s Imam Council, told Anadolu Agency that the bill is against the country’s constitution.
“The country already has a huge population. This bill is against the constitution and there is no need to give citizenship right to those people who are coming from outside and living illegally in the country,” he said.
Urmilesh, a senior Indian journalist and political commentator, believes the bill will hamper the political interest of BJP in the northeast.
“But they don’t bother so much about northeast for now because there are not many seats from that region [for the general elections]. The only interest they had was in state elections which are already over,” he said.
The BJP however maintains its has popular support in the region.
“Those who are agitating on the ground are being misguided by the people…other political opponents are trying to misguide the people…..If people are against us, then they would never vote for us,” local BJP leader Pabitra Margherita told Anadolu Agency.
Experts also say the amended citizenship bill will affect the update of National Register of Citizens (NRC).
The NRC list is unique to Assam, which was first tabulated in 1951, four years after India’s independence from British rule, to distinguish Indian citizens from illegal migrants from bordering Bangladesh.
After updating the list in 2015, the government last year left out 4 million people.
Some 3 million people have contested the claim and the Supreme Court in monitoring the process.
“The current update of the National Register of Citizens (NRC) will also be rendered meaningless if a section of illegal migrants who are left out of the list are granted citizenship based on religion,” said Wadud. (AA)

domain: news


text:
Poll Archive: Which social media (or method) do you use to follow THF updates?
I don't follow THF on social media 35%
Disclaimer: All publicly recognizable characters, settings, etc. are the property of their respective owners. The original characters and plot are the property of the author. No money is being made from this work. No copyright infringement is intended.

domain: literature


Style: Melodic Death Metal |
Year of birth: 1995
Views: 5099. Updated: 14 May, 2005 by Project Moderators
Members of Urkraft
| Current Members:|
Thomas Stromvig Pedersen (Founder). In Band at: 1995 - . Role: Guitars, vocals
Thomas Birk (Guest). In Band at: 2000 - . Role: Guitars
Side projects: Cacophony
Jeppe Tander (Founder). In Band at: 1995 - . Role: Bass
Jeppe Eg (Guest). In Band at: 2002 - . Role: Keyboards
Side projects: Blazing Eternity, Cacophony, Manticora
Mikael Skou Jorgensen (Founder). In Band at: 1995 - . Role: Drums
R. Mork (Founder). Period: 1995 - 2000. Role: Guitars
|Demo: Verden vil Bedrages (1996) Tracklist.|
Comment: First demo-tape recorded in Svendborg, Denmark
Demo: 2000 (2000) Tracklist.
Comment: Recorded at Black Tornado, summer 2000; Produced by Lars Lundholm
Demo: 2001 (2001) Tracklist.
Comment: Recorded in the basement with Crooked Studios, spring 2001
Demo: 2002 version 1 (2002) Tracklist.
Comment: Recorded in the basement with Crooked Studios, autumn 2001
Promo: 2002 Version 2 (2002) Tracklist.
Demo: Primordial Promo 2003 (2003) Studio: SoundZone Studio. Tracklist.
Comment: Produced by Lars Schmidt, mixed and mastered by Jacob Hansen, spring 2003
nothing: Eternal Cosmic Slaughter (2004) Studio: Antfarm. Tracklist.
Comment: Produced, engineered, mixed and mastered by Tue Madsen. All music by Birk & Thomas, arranged by Urkraft; Lyrics on 1, 3, 4, 8 by Birk; Lyrics on 2, 5, 6, 7 by Mikael; Lyrics on 9 by Thomas
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Reduce CO2 emissions
To optimize the operation of individual properties, upgrading of technical facilities including heating and lighting has our greatest attention. We conduct thermal insulation as long as the properties can preserve the identity. It is our sustained goal to reduce CO2 emissions. In connection with the renovation of buildings, construction waste will been deposited, so it can be recycled later. Recently we bought an economically electric mini car for transport in the inner city.