Is a flexible formwork for confining concrete lift core wall on the construction of high rise buildings that saves time, labour and cost.
Allows large area of concrete wall to be formed easily, precisely and completed within a limited time. By using HIFORM wall form system, high quality concrete walls can ve obtained by none skill labours with minimum work force.
Where complicated sections are to be constructed and the standard formwork cannot be applied, Plywood face wallform will be supplied to complete the work.
Utilies lipped channel or RHS with column waters in two parts - male and female which are connected by tension pins to form square and rectangular columns.
Is a horizontal support beam specially designed with high strength for an overall rationalization of concrete slab construction.
Is suitable for high-rise construction. Relatively quick turn-around time for striking, flying and reseting to enable it to achieve a shirter construction cycle with minimum manpower.
Is a cell system made entirely of steel and waith all its load-bearing parts possessing great rigidity.
Is a flexible formwork for confining concrete wall on yje construction of silo that saves time, labour and cost.
Different types of metal form are fabricated for the constructions of walls, columns, beams and slabs. They are accurate, durable and cut down working time and cost of construction. They are supplied in many forms, shapes and sizes according the job requirement. It minimize usage on crane and all panel can be hand held easily.
Address: Lot 1084 Jalan Budaya Off Jalan Kapar Batu 3 1/2 Rantau Panjang 42100 Klang Selangor Darul Ehsan Malaysia
Telephone: +603-3291 7353
Fax: +603-3291 7290
Copyright 2017 HIFORM (M) SDN BHD. All Rights Reserved.


text:
Over at National Review yesterday, Ramesh Ponnuru--a conservative I respect and enjoy reading--criticized Jonathan Chait's column
on Bill Kristol. In particular, Ramesh said, Jon has unfairly seized upon a particular passage in Kristol's recent editorial about liberals and the troops. It's the passage where Kristol wrote:
Having turned against a war that some of them supported, the left is now turning against the troops they claim still to support. They sense that history is progressing away from them--that these soldiers, fighting courageously in a just cause, could still win the war, that they are proud of their service, and that they will be future leaders of this country.
Ponnuru acknowledged that this is "an overheated and unfair statement," but went on to suggest that "Perhaps Jonathan Chait has made overheated and unfair statements from time to time? I don't think it would be reasonable to take one of those statements and use it to create a theory of his essential thuggishness."
Well, sure. Jon has made overheated and unfair statements from time to time. So have I. So has everybody. But this wasn't just a throw-away line. It was the premise of the whole article. That is why, I imagine, it appeared under the headline "THEY DON'T REALLY SUPPORT THE TROOPS."
Yes, the editorial spent much of its time discussing the Scott Beauchamp articles--which, as Jon freely admitted, were the subject of some legitimate questioning. But Kristol used those stories to make a broader point about TNR's motives--and those of war critics generally. That broader point is what attracted Jon's scrutiny.
And rightly so. Kristol's outburst was hardly out of character, after all. In yet another signed editorial that ran in February, Kristol asked whether Republicans would "desert the troops" by supporting a non-binding resolution against the surge--a measure he termed "anti-surge, anti-Petraeus, anti-troops, and anti-victory resolution." ("Anti-surge," "anti-troops," apparently they mean the same thing...) One week later, after that resolution passed the House, he wrote that "The national Democratic party has become the puppet of antiwar groups. These groups do not merely accept--reluctantly--American defeat in the Middle East. They seek to hasten it. Some seem to welcome it." And then, just a few weeks ago, there was the cover Jon mentioned in his column--the one featuring an American soldier, viewed through what looks like a gun-sight, and the headline "DOES WASHINGTON HAVE HIS BACK?"
In these same articles, Kristol also made more level-headed arguments about policy. He has every right to express those opinions and, lord knows, he can speak to the issue with more authority than I can. But I know enough to spot demagoguery when I see it. And that's precisely what we have here. Over and over again, Kristol has tried to disqualify his political opponents from debate by suggesting they don't support the troops, don't want a strong America, or don't have firm moral principles.
The approach reminds me a lot of Zell Miller's unhinged speech at the Republican Convention in 2004--which, as I said at the time, bore more than passing resemblance to the speeches of a certain Wisconsin senator who served during the 1950s. The only difference is that, unlike Miller, Kristol still has respectability in Washington. Kinda makes you wonder why.

domain: politics


text:
Watch your pizza being made in front of you at Brother's Pizzeria and enjoy it hot. Specialty Pizzas on offer include Brother's Supreme, Buffalo Chicken Pizza and more. Cold subs like Roast Beef, Turkey, Provolone and hot subs like Veal Parmigiana, Cheesesteak, Grilled Chicken Fillet are also worth a try. Sides like Buffalo Wings, Jalapeno Poppers and Garlic Bread are also available along with...
Sicilian & Regular Pizza
Whole Pizza Pies
Hot and Cold Heros
Calzones & Heroes
Calzones & Rolls
Honestly the best pizza around. They have so much business they don't even deliver. Pickup only. If you want the best you got to get off your butts and go get it.
» Read More:
As you drive up forest ave, you will see Brothers Pizza sitting on the corner like a food oasis, the whole scene looks straight out of a modern Italian movie!, did De niro and Pesci just walkout with a pie?
When you walk in your welcome by a glass case full of old world style pizzas, like the grandma, Sicilian,...
Love their pizza. It's a great go to pizza after coming home from bar and clubs. They sell individual slices.
These are true Italians making n selling pizza, what more authenticity do you want.
Don't miss this place or else your missing out on true authentic pizza.
Brother's Pizza can be found at Port Richmond Ave 750. The following is offered: Carry Out & Take Out. The entry is present with us since Sep 9, 2010 and was last updated on Nov 14, 2013. In Staten Island there are 79 other Carry Out & Take Out. An overview can be found here.
you tried the rest...now try the best

domain: food


text:
LaDainian Tomlinson and the San Diego Chargers were a force in the mid-2000s, as the future Hall of Famer became a superstar. But one thing apparently still stick’s in Tomlinson’s craw: the 2006 season when the Chargers went 14-2 and lost to the New England Patriots in the divisional round of the playoffs 24-21.
Tomlinson believes the Chargers would have won the Super Bowl that year if they had Drew Brees under center, not Philip Rivers (who was in his first year as a full-time starter). Chris Mortensen relayed the following quote from Tomlinson:
“If Drew had been our quarterback [in 2006], we would have won the Super Bowl. There were a lot of circumstances that were in play — obviously, Drew’s injury right at the end of the (2005) season and the front office wasn’t willing to make the (financial) commitment to keep him.
“Philip hadn’t played enough to get it done at that time.”
Tomlinson’s argument certainly has merit. While in the long run the decision to go with the younger, cheaper (at the time) Rivers was probably the best decision for the Chargers business-wise, that 2006 season might have ended differently with the experienced Brees at the helm.
That said, you can’t argue too much with a quarterback who guided his team to a 14-2 record. Rivers was good in 2006 as he completed 61.7 percent of his passes for 3,388 yards, with 22 touchdowns and nine interceptions. He had a passer rating of 92.0 and a Total QBR of 69.8, both solid marks. But maybe his lack of experience in big moments held the team back in the postseason.
Tomlinson won the MVP award following a record-breaking season in 2006 but it’s clear he feels he and the team came up short.
Following the 2006 season the Chargers’ front office made the asinine decision to fire head coach Marty Schottenheimer[autotag] and replace him with Norv Turner. While the team made the AFC title game during the 2007 season, things were never the same. That 2006 season was San Diego’s best-ever chance for a title, even better than their run to the Super Bowl in 1995

domain: sports


text:
Preheat the broiler. Place the dulse and wakame in separate bowls, and cover each with cold water. Set aside until rehydrated and softened.
Place the bell peppers on a baking sheet, cut side down. Broil 2 inches from the heat until black and charred. Transfer the peppers to a bowl, and seal with plastic wrap. Set aside for 10 minutes or until they are cool enough to handle. The skins will easily scrape free with the back of a knife. Discard the charred skins, slice the pepper halves into thin strips, and return them to the bowl.
Drain the dulse and wakame. Spread out the wakame, and cut away any tough veins. Coarsely chop the wakame and dulse, and add them to the bowl with the peppers along with the cucumber, radicchio, and endive.
In a small bowl, whisk together the vinegar, lemon juice, jalapeno, garlic, lemon zest, 1 teaspoon salt, and pepper. Add the oil in a steady stream, whisking to emulsify.
Pour the dressing over the vegetables, and toss to coat. Refrigerate salad for 30 minutes.
Season salad with additional salt and pepper to taste. Sprinkle with cilantro and serve.


In the 2014 Kentucky General Assembly session, several critical issues are being considered but two rise to the top of my list of concerns for families in our state.
Subscribe to read the full story
Current Subscribers - Activate Now
Already subscribe to the Evansville Courier & Press or The Gleaner? Unlimited access to CourierPress.com, TheGleaner.com and the Courier & Press smartphone and tablet apps is included with your subscription. All you need to do is ACTIVATE now!
New Subscribers - Subscribe Now
Want to keep reading?
We now offer Premium and Digital Subscriptions. Subscribe now and select how you want to keep up-to-date on local news, reader comments, photos, videos, blogs and more.


text:
WIN with Golf Ireland Belfast Golf Event 2022
Do you play Golf? Then why not come along to the Golf Ireland, Golf Promotion Night in association with Fáilte Ireland in The Europa Hotel, Belfast on Tuesday 22nd March from 6pm – 9pm.
It's a FREE event to attend with golf simulator, refreshments and prizes for all ages!
All this week on Q Radio Drive, we have teamed up with Golf Ireland in association with Fáilte Ireland to give lucky golf enthusiasts the chance to win one of five daily prizes. All you have to do to win is tune into Q Drive with Ibe Sesay, 4-7pm Monday to Friday and play the game ‘Who's in the Bunker?'.
Monday: 4 Ball Voucher for Sandy Hills Course located in Donegal at Rosapenna Hotel & Golf Resort
Tuesday: Overnight stay midweek in an Executive Suite for two with breakfast the following morning at Harvey's Point Hotel
Wednesday: 4 Ball Voucher for Roganstown Hotel and Country Club, Co. Dublin
Thursday: 4 Ball Voucher for Donegal Golf Club, Laghey, Co. Donegal
Friday: 4 Ball Voucher for Nuremore Hotel & Country Club
The Republic of Ireland offers world-class golf experiences wherever the signpost points you, whichever road you take. Go beyond the golf course, look deeper and you’ll find moments that last a lifetime.
If you want to play the best courses, if you enjoy club history or you’re a design aficionado – or maybe you just want to have a little fun and soak up some scenery - then you have many shades of green to choose from.
Try the revered links of the south west, or those around Dublin or Carne, a Co. Mayo links within striking distance of Enniscrone, Strandhill and Co. Sligo golf clubs.
County Donegal possesses links by Tom Doak (Rosapenna St Patrick’s Links), and another by Gil Hanse (Narin & Portnoo). Add these to names like Old Tom Morris, Harry Colt and Ireland’s Pat Ruddy, and you have something that offers all types of history and challenge. The beauty of the Wild Atlantic Way coastline is matched by Donegal’s 14 links courses. Whichever you play you’ll enjoy the sound of the ocean and the poetry of a golf swing carving through the wind.
In the Republic of Ireland you'll find courses that ramble across mountainsides, dive through valleys, wrap around lakes and embrace rural tranquillity. Take Co. Wicklow. The European Club is a bucket list links but just to the south the short, sharp links at Arklow packs a punch. To the north the neighbouring Blainroe and Wicklow golf clubs are very different seaside courses. Nearby is Druids Glen, home to four Irish Opens and the best set of par threes you’ll ever play. Inland is the charming Woodenbridge, lost in a valley of deep forest, while the Paul McGinley-designed Macreddin erupts nearby in another valley. The county is home to 20 courses.
And that’s just a taster of what’s on offer around the Republic of Ireland. Make sure you come along to the Golf Ireland, Golf Promotion Night in association with Fáilte Ireland in The Europa Hotel, Belfast on Tuesday 22nd March from 6pm – 9pm to find out about lots more!

domain: travel


text:
Holiday 3-Pack: Mini Plush Dog Toys
Zippypaws Miniz Donutz
ZippyPaws create fun, colourful toys and accessories of the highest quality. At their core, ZippyPaws always delivers the best for your dog. Each item is created is by sketching every design, hand-selecting the perfect materials, and testing all finished products with a furry testing team.
Spoil your pet with Miniz Donutz. Suitable for small to medium sized dogs, featuring three soft and stuffing free Donutz containing crescent-shaped squeakers for double the entertainment.
- 3 plush donut dog toy set
- Features stuffing-free interior with crescent-shaped squeakers in each donut
- Great gift for multi-dog homes or to spoil a single dog
- Ideal for small to medium dogs to enjoy
- Fun and interactive
- Approximately 15.2cm x 11.4cm x 5.1cm

domain: pet


text:
NEW YORK (AP) — Fresh off his Broadway run, New Jersey’s Bruce Springsteen looked West for inspiration on a new album he’s planning to release on June 14. The 13-song disc is his first new studio album in five years.
Titled “Western Stars,” Springsteen said Thursday the album draws some inspiration from southern California pop records of the late 1960s and early 1970s.READ MORE: Need A Mail-In Ballot? Deadline To Request One Is Tuesday In Pennsylvania
He’s releasing a song and video called “Hello Sunshine” after midnight on Thursday.
Springsteen wrapped up a 236-show Broadway run last December.READ MORE: Philadelphia Weather: Flood Warnings Still In Effect For New Jersey Counties As Storm Moves Through
Springsteen recorded it primarily at home in New Jersey, with additional work in California and New York. Ron Aniello produced and plays on the album, which also has contributions from Springsteen’s wife Patti Scialfa, Jon Brion, David Sancious, Charlie Giordano and Soozie Tyrell.
“This record is a return to my solo recordings featuring character-driven songs and sweeping, cinematic orchestral arrangements,” he said. “It’s a jewel box of a record.”
Among the song titles are “Hitch Hikin’,” ”Sleepy Joe’s Cafe,” ”Chasin’ Wild Horses” and “There Goes My Miracle.”MORE NEWS: Police: 18-Year-Old Zion Davis-Shelton Fatally Shot In Wilmington, Delaware
(Copyright 2019 by The Associated Press. All Rights Reserved.)

domain: artistic


text: "I will be your shield. However, if you believe in Alfred Lord Tennyson's telling of the story, Nimue is more evil, Merlin's entrapment is "voluntary," and the love flows from Merlin to Nimue — not the other way around. Nimue soll es zurück zum Zauberer Merlin bringen. She replaces Merlin in court as the King’s main source of magic. Richard Wilbur referred to Vivien as “a creature to bewitch a sorcerer”, and even though Merlin could foretell and foresee his captivity, he was unable to stop it or overcome his enchantment with Vivien. Merlin believes Nimue to be dead, and is racked by grief as Vortigan turns to him. Die Teenagerin Nimue (Katherine Langford) besitzt eine geheimnisvolle Gabe. And I vow on this day that the Fey will live free again!" At the end of the tale, she carries the wounded Arthur to Avalon. You may be looking for the episode "Nimue." In Thomas Malory’s Morte D’Arthur, Nimue is a goodie. ― Nimue to the Fey people[source] Nimue is the protagonist of Cursed. 4. She is a Fey with the power to control plant life around her. And, by the gods, I will be your sword. Vivien, sometimes called Nineve, Nimue, or Niniane is best known as the woman who sealed Merlin in a cave or a tree and put him under a spell. Eingebettet in eine klassisch-abenteuerliche Coming-of-Age-Geschichte ist der Roman mit Miller-typischen schwarz-weißen, aber auch kolorierten Illustrationen. Auf ihrer Suche nach Merlin macht sie Bekanntschaft mit einem jungen Söldner namens Artur. Nach dem Tod ihrer Mutter soll sie ein uraltes Schwert zu Merlin (Gustaf Skarsgard, 39) bringen. This article is about the character Nimue. Unexpectedly, at this point, a healthy Nimue rips out Vortigan's heart and reveals that she had drunk from the Holy Grail, desiring power and immortality like Merlin's, and thirsty for vengeance against Vortigan.

domain: literature


text:
Electric Six have launched their latest Kickstarter. Entitled ‘A Very Electric SiXmas & Chill Out!’ it features two discs: the first will be a 12-track Christmas album and the second an acoustic live album.
Backer bonuses include a second karaoke disc, t-shirt, Christmas stocking, personal drunken phone call from DV and much more.
Electric Six is proud to announce its latest Kickstarter campaign today. Entitled “A Very Electric SiXmas & Chill Out!,” this is a two disc set comprised of two very different sets of material. The first disc is E6’s first ever attempt at an old chestnut, the holiday record. Our cover of “You’re A Mean One, Mr. Grinch” for The Onion’s AV Club several years ago has been a holiday favorite with the Crazies and we’ve decided that it’s time for an entire record filled with soon to be magical songs and memories associated with the holiday seasons. We’ve indicated a few of the songs we’re going to do in the campaign, but there’s room for many, many more to come. Some of those will be selected by us and some by you, the Crazies, as part of the campaign.
The second disc, the “Chill Out!” record, is another new venture for us. Dick Valentine has been doing live acoustic performances for several years, and he often includes E6 songs in his set. We’ve decided to perform and record a live acoustic set by the entire band. We’re doing it on March 21, 2018 at Small’s in Hamtramck, Michigan (DaVe’s bar/club) and we think it’s going to offer an entirely different perspective on the songs the band opts to do. Again, as with the holiday record, we’ve indicated a few songs which will be part of the set in the campaign, but there will be many more to come via our selections and those of the Crazies. Believe it or not, this may be one of the most challenging things the band has ever done, but Electric Six is ready to step up and face the (non-electrical) music. Now, it should be understood that delivery of these records and all the associated packages will not happen as part of the current holiday seasons. It takes poor old Electric Six awhile to pull everything together, so you’ll get everything in October of 2018, in time for NEXT year’s festivities. The best things in life are worth the wait.
As you look through the campaign, you’ll see the usual wide variety of items on offer. Lots of old favorites are once again available, but we have a few new ones on offer as well, mostly those with a holiday theme. Dick Valentine-created holiday cards! A Dick Valentine stained glass night light! A customized E6 Christmas stocking! Check out all the great packages at the link below. Happy Holidays to everybody!!!
Click here to find out more and to take part.
Via a post on the band’s official Facebook page on 6th July the band have announced their upcoming album will be called ‘How Dare You’.
On 19th July they uploaded the artwork and track listing. You can also now pre-order the album online.
More on this as we get it. The album is due out this autumn.
The first of two Kickstarter videos from the recent ‘You’re Welcome’ campaign has been released online, via the band’s Facebook page.
You can view the video below which sees two Dick Valentine’s, an old fashioned feel and an, er, interesting prop.
A post on skiddle.com to announce an accompanying live date has revealed the title of Valentine’s fourth solo studio album.
come one, come all to the intimate and festive occasion that is the dick valentine solo acoustic experience! experience a seven-or-eight-times-in-a-lifetime chance to be up close and personal with an american music icon. it’s a low pressure, good time affair as the electric six frontman strips down all your favorite songs from the electric six catalog, plays original solo material and swims in a sea of requested good-time covers.
this tour will be your chance to reach directly into dick valentine’s bag and be one of the first to buy a copy of his latest solo work “robocalls.” “robocalls” is valentine’s fourth solo album and there can be no doubting it will go down as his magnum opus. imagine mac demarco being spliced with gordon lightfoot…and you’ve got two canadians being fused into one. in the background during the splicing procedure, you might hear a trace of “robocalls” emanating from the anesthesiologist’s ipod.
so you know the drill….drop everything and come see dick valentine behaving as a cornered animal as he comes face to face with you, the fans. an night of merriment and frivolity awaits! come see the american troubadour dick valentine take the stage in your town for one unforgettable evening. you will not see electric six in 2017 so this is the next best thing!
According to a release on his Patreon page the album has mostly been recorded and includes a track called ‘The Room Where You and I Will Make Out’.
Nine tracks were done the other day at Figure 8 studios in Brooklyn (three doors down from Hood Hing). The other three tracks are being done in Michigan with The White Wolf. Gonna be a good one!
Other dates on the tour include York, Hull, Newcastle and Cardiff.
The lead singer of Electric Six has joined ‘Patreon’, a crowdfunding service where creators offer subscriptions that fans can pay for in return for content. He promises on his page a selection of covers for supporters.
You can find out more here.
‘Randy’s Hot Tonight!’ has been voted as the next song to be included in the ‘You’re Welcome’ Kickstarter, beating ‘Pulling the Plug on the Party’ to the top spot.
Randy’s Hot Tonight! – 9 votes
Pulling The Plug on the Party – 7
Bite Me – 5
Countdown To The Countdown – 2
This follows ‘I Invented the Night’ and ‘Dance Pattern’ added on previous stretch goal votes. An additional stretch goal – not based around a vote – is expected today (Sunday).
Electric Six have launched their fourth Kickstarter campaign for a new live album recorded in Oxford in April 2017.
The album, entitled ‘You’re Welcome’, will be recorded at the Oxford Academy on 22nd April 2017 and released to Kickstarters alongside a range of incentives including a t-shirt, poster and karaoke disc, alongside other one-offs such as choosing a track to be performed live for the album, a special encore and a tarot reading from lead singer Dick Valentine.
The 2-disc set will include an album of covers and the live album, which will consist of songs that didn’t feature on live album ‘Absolute Pleasure’ or DVD / Blu-Ray ‘Absolute Treasure’, some not having been played live before.
Alongside backers being able to pick live songs, the following five tracks have been confirmed for inclusion on the live disc:
- Newark Airport Boogie
- I’m The Bomb
- Night Vision
- MC Sucka DJ
- Rock and Roll Evacuation
Alongside the usual offers there is a chance for backers to become part of a subscription network where they get a special track every three months, which may be a demo, remix of fully finished track preview.
The Kickstarter campaign is looking for $30,000 and had already reached $18,000 within twenty-four hours of its launch.
You can also buy tickets for the event at www.ticketweb.co.uk/event/DEH2204Z
You can now listen to two new songs from the upcoming latest Electric Six album online.
By the power of transcription, still a little incomplete, you can also read the lyrics to ‘The Number of the Beast‘ and ‘I’ll Be In Touch‘.
According to this post on Amazon the new album will be called ‘Fresh Blood For Tired Vampyres’, which has now been confirmed on Electric Six’s official Facebook page.
Due for release on October 7th here is the press statement:
Eternal life can seem like an eternity. Ask any vampire. The continuous march of sun ups, sun downs, transformations of form, seductions, cape fittings & exsanguinations. Eventually it all just becomes an endless, tired routine. It all seems so exciting & so sexy to those of us who operate knowing we have limited time. But ask any vampire about the downside of eternal life, & you won’t be surprised to hear tales of binge eating garlic bread just to feel the hurt, or of the occasional dangling of a wooden stake just over the center of the rib cage. Electric Six knows all about eternal life. Electric Six has been around forever & it can never die. That’s lovely, but it’s also very tiring. Fresh Blood For Tired Vampyres is the new release by Electric Six on Metropolis Records. One listen & you will immediately understand that the sexiest vampires are urban vampires. Where E6 has dabbled in dance, hip-hop & R&B in the past, Fresh Blood is the whole enchilada. It’s thirteen songs designed to make the listener interested in smooth & nasty fuckin’, the way they do it in the city. From the Grandmaster Flash-inspired Number Of The Beast to the super smooth tour of the NYC outer boroughs Mood Is Improving, the listener finds himself immediately deposited into an urban drop zone with hustlas & dickblockas coming from behind every corner. The radio-ready pop hits I’ll Be In Touch & Dance With Dark Forces are the tracks that get the listener off the street & into the club. & it would not be an Electric Six album without an epic closer, that being the beautiful & haunting Spacewalkin’, the ballad that assures the listener that the vampire has now fed & will live a thousand more years, albeit in outer space. Electric Six changes more frequently than change itself, but ultimately this just means they’re never gonna put out the same album twice. Fresh Blood for Tired Vampires is poppy & smooth, nasty & raw…& oh so life affirming, especially if you are undead
More as we hear it.
Below are the confirmed tracks for the upcoming new album:
1. Acid Reducer
2. Number of The Beast
3. Mood Is Improving
4. I’ll Be In Touch
5. Lottery Reptiles
6. Dance WIth Dark Forces
7. Skin Caboose
8. My Dreams
9. Greener Pastures
10. I Got The Box
11. Lee Did This To Me
12. The Lover’s Pie
13. Space Walkin’


text:
| Jun 11, 2013
| 9:48 AM
Joseph Marinaro had a dream: He was going to be a great singer, like his heroes Frank Sinatra and Mario Lanza.
But the talented young crooner from Buffalo, N.Y., got married and had 10 kids, set aside his musical ambitions and laid asphalt to support...
| Jun 6, 2013
| 12:49 PM
If jazz listeners ruled the world, saxophonist-composer Wayne Shorter's forthcoming 80th birthday – on Aug. 25 – would be deemed a national holiday.
That doesn't seem likely at this point, so Chicago will celebrate the occasion a bit early...
| May 28, 2013
| 11:40 AM
The great flutist-composer Nicole Mitchell moved to Chicago – and launched her international career here – in 1990.
More than two decades later, in August of 2011, she left the city for California, yet it's still impossible to think of...
| May 16, 2013
| 12:39 PM
Though its population hovers at just 8 million, Israel has become a powerhouse in nurturing high-caliber jazz talent.
Chicago concertgoers are well acquainted with Anat Cohen, the uncommonly versatile Israeli clarinetist who's based in New York and...
| May 14, 2013
| 9:55 AM
Let's raise a glass to Bob Koester, the single-minded Chicagoan who this weekend celebrates a remarkable anniversary: 60 years of producing jazz and blues recordings.
Koester's Delmark Records may not be the biggest indie in the country – or...
| May 9, 2013
| 9:10 AM
Chicago jazz musician Miguel de la Cerna likes to refer to himself as "your basic piano player, commercial musician," but that modest self-assessment greatly understates the case.
De la Cerna proved the point dramatically last year, when he partnered...
| May 7, 2013
| 9:28 AM
Bluesman James Cotton has lived the kind of life they make movies about.
He toiled on a Mississippi plantation from earliest childhood, found himself orphaned at age 9, practically was raised by blues master Sonny Boy Williamson II and flourished as a...
| May 1, 2013
| 6:54 AM
Drummer Jack DeJohnette, trumpeter Wadada Leo Smith, pianists Jason Moran and Robert Glasper and saxophonists Donald Harrison, Charles Lloyd, Jimmy Heath and Rudresh Mahanthappa will headline the 35th annual Chicago Jazz Festival, running Aug. 29...
| Apr 23, 2013
| 10:43 AM
Imagine what might happen if some of Chicago's most inventive jazz musicians embarked on a multi-year collaboration with their counterparts in France.
Just think of the music that might result, the insights that could be shared, the audiences that would...
| Apr 19, 2013
| 7:11 AM
For more than three decades, the Palestinian oud virtuoso Simon Shaheen has relentlessly championed his soft-spoken instrument across America and beyond.
He has penned solo works, pieces for oud with wind quintet and string quartet and compositions...
| Apr 18, 2013
| 2:44 PM
For 511 days, they hid underground in Ukrainian caves while fellow Jews were being executed by machine gun up above.
The odds that they would survive the Holocaust were slight, for they were dependent entirely upon themselves for finding scarce food...


text:
UBER Part Time Driver - Earn Money on Your Schedule
Call Center In Texas Is Hiring
$22 / HR. NO EXPERIENCE NEEDED. AIR LINE HIRING.
Owner Operators Needed. Non-forced Dispatch. More Home Time.
High-Earning Sales Career Training Provided
Now interviewing - Work from home
Food Service Line Specialist
Sales Consultant - Full Time - Venus
Carhop/Skating Carhop (Server)
Two positions, LVT and Veterinary Office Reception.
Assembly Production Worker
Assistant Manager - Restaurant
Customer Service F/T or P/T No Exp. required
Customer service home office
Store Team Member
Target Warehouse Associate
F/T - P/T Reps Wanted
Warehouse Jobs-Midlothian TX-$11.50/hr
Find the career you've been looking for.
The Maypearl, TX Job Market
Aiming for a career in Maypearl, TX? Maypearl, TX contains a population of approximately 1,000, according to 2013 BLS data. The median residential take-home pay number of Maypearl, TX is approximately $56,000, as indicated by BLS statistics citations from roughly 2013 Maypearl's cost of living, as of 2013, was something like 84 out of 100, which is lower versus the nationwide average.
If you might be seeking out work opportunities throughout the city, live in another place and searching in other towns and cities, or if you are searching for areas worthy of moving to which also have good job possibilities, it is worthy of considering the state of unemployment in the city. Since 2014, Maypearl had an unemployment rate of somewhere around 5.5%. This is higher than the United States average unemployment rate, from late 2015. Ideally this can aid you for creating a bit more educated assessment with regards to your prospects in this city.

domain: geography


text:
WESLEY KIKUCHI is the Founder, Wellness Director and Nutraceutical
Specialist for the Institute of Anti-Aging and Longevity of Carson City,
He is the President of Physicians Nutrition Network, an educational
forum for licensed health professionals to learn more about Complementary
Therapies as part of their practice. He is also President of www.ImmuneSolutions.net,
a website dedicated to the implementation, quality, reimbursement and
education of health professionals using Nutraceuticals as co-therapy
Wesley has over 10 years of professional experience in nutritional supplementation
as a consumer health advocate and 8 years in high technology medical
and diagnostic test equipment. He is a business health consultant to
Medical Groups, Hospitals, Fitness Clubs and Allied Health Professionals
interested in integrating Nutraceuticals.
He is also the Sports Nutrition Director for the Athletic Dept. of Calumet
College in Indiana and the past Sport Nutrition Specialist for the Arthritis
Foundations Marathon Team in training.
Wesley gives seminars around the U.S. on 30 different health education
topics to a variety of audiences such as AARP, Senior Friends and the
American Lung Association.
He has a local cable TV show in Douglas County, Nevada called Wellness
In Practice, which focuses on consumer awareness on Complementary
and Alternative Therapies. The show is also syndicated on cable access
in Carson City and Reno, Nevada.
He is a regular on radio shows such as Your Choice, Your Health
and Health Matters on the importance of nutrition, supplementation
and integrative medicine.
Wesley will speak on Supporting The Immune System Against Disease.

domain: health


text:
How to write the perfect crime, according to Agatha Christie
340,081 views | Jamie Bernthal • TED-Ed
With almost 100 mystery novels, each one a cleverly constructed puzzle box of clues, misdirection, and human drama, Agatha Christie is the best-selling novelist of all time. Her eccentric detectives, clever clues, and simplified suspects have stumped countless readers over the last century. So, how did she craft these perfect crimes? Jamie Bernthal dissects the writing of Agatha Christie. [Directed by Anantha Krishnan, Nikhita Prabhudesai, narrated Addison Anderson, music by Salil Bhayani, cAMP Studio].

domain: entertainment


text:
4 slices red onion
1 red bell pepper, seeded and quartered
1 (12 ounce) eggplant, cut into 1/2 inch-thick pieces
2 Tbsp olive oil, divided
1/4 cup chopped fresh flat-leaf parsley
1.8 tsp kosher salt
1 (8 ounce) container plain hummus
4 (1.9 ounce) whole-grain flatbreads
1/2 cup crumbled feta cheese
1. Heat a large grill pan over medium-high heat. Brush onion, bell pepper, and eggplant with 1 Tbsp oil. Add onion and bell pepper to pan; cook 3 minutes on each side or until grill marks appear. Remove from pan; coarsely chop vegetables. Combine vegetables, remaining 1 Tbsp oil, parsley, and salt; toss to combine.
2. Spread 1/4 cup hummus over each flatbread, leaving a 1/2 inch border around edges. Divide vegetables over each flatbread; top each serving with 2 Tbsp cheese. Roll up wraps, and cut diagonally in half.
Yield: 4 servings
Nutrition information per serving: 356 calories, 23 g fat, 3 g saturated fat, 17 g protein, 35 g carbohydrate, 15 g fiber, 788 mg sodium
Source: Cooking Light


text:
What a wonderful stay with the best hosts, Bridgette & James. Nothing is too much trouble for this fun couple. We were a group of 5 friends sharing the family apartment. Spacious and well equipped as well as serviced every day which was a joy. Large braai area for get togethers. Villa Ayr is situated close to the beach and... more
You have no recent hotel searches.


Tanzania is the 30th largest country in the world and 13th largest in Africa with a population of about 45 million. It is about the size of Texas and Colorado combined or approximately the size of France, Germany and Switzerland combined.
Tanzania is located in East Africa and is comprised of the mainland and Zanzibar archipelago. It is surrounded by Kenya, Uganda, Rwanda, Democratic Republic of Congo, Zambia, Malawi and Mozambique.
Tanzania is a country that has been endowed with some of the most beautiful natural wonders in the world. In fact, three of the seven natural wonders of Africa are in Tanzania: Mount Kilimanjaro, Ngorongoro Crater and the Serengeti Migration. Tanzania boasts the highest peak in Africa, Mount Kilimanjaro; the deepest lake in Africa, Lake Tanganyika; and the largest lake in Africa, Lake Victoria.
Tanzania has a tropical climate. The climate varies from tropical along the coast to temperate in the highlands. Due to its proximity by the equator, the cooler months occur while it is summer in the northern hemisphere. The coastal regions are warmer and more humid with temperatures ranging between 25 and 31 degrees centigrade ( 77 to 87 degrees fahrenheit) between November and February and dipping below 25 degrees in the coolest months between May and August. In the highlands, temperatures range between 20 and 23 degrees centigrade (68 and 73 degrees fahrenheit) throughout most of the year and only dipping a couple degrees below 20 during the months between May and August.
There are two rainy seasons; the short rains are generally from October to December, while the long rains last from March to June.
Tanzania Wildlife Safari can be visited year round; however, the most favorable time to travel is outside the rainy and when the climate is cooler, between June and September. Beach side locations like Zanzibar can be fine to visit during the hotter months of November to February, when ocean breezes make the high temperatures bearable although humidity can still be high.


text:
Less than a week before the midterm elections, US House Rep. Raúl Grijalva (D-Ariz.) released a report detailing how the US House Committee on Natural Resources, on which he has served for 14 years, stacked its hearings with industry interests. “Under Republican leadership,” he wrote, “hearings have disproportionally included witnesses who pad their profits by degrading public lands.”
Now that Democrats have won a majority in the House, Grijalva will have his chance, as the committee’s new chairman, to change the direction of the governing body that oversees federal lands and energy and water resources. Grijalva’s committee will also oversee and investigate the Interior Department, employing the system of checks and balances that Grijalva thinks his predecessors neglected.
Last week, High Country News spoke with Grijalva about his priorities and what his leadership could mean for climate change policies and resource management in the West. This interview has been edited for clarity and length.
High Country News: As you assume chairmanship, what do you hope to do differently than [outgoing chairman] Rep. Rob Bishop (R-Utah)?
Raúl Grijalva: We have an opportunity to take this committee and its priorities and its policies and legislative initiatives and steer it in a different direction. Under our jurisdiction, we have issues that have to be dealt with—tribal sovereignty, education, health care, historical and cultural resource preservation.
The other issue is climate change. It touches every issue that we deal with, and the fossil fuel extraction industries are making such a rush for resources in our public lands. This administration, in two years, has made every effort to suppress science and dumb down the issue of climate change. We want to elevate that again to the status it deserves in decision making.
HCN: How will you do that?
RG: We will begin to look at ways in which our jurisdiction can help mitigate the effects of climate change. We’ll do that legislatively, by holding hearings and introducing policy initiatives. My committee will revisit all of the rules that have been changed by this administration that have to do with climate change and science, the Endangered Species Act, the Wilderness Act, and responsibility over our federal waters and waterways. The list goes on and on.
HCN: How effective can you realistically be in changing the direction of the committee?
RG: The prerogatives of the current administration will limit our authority. With this administration, we have had no oversight hearings. Our request for accountability and inquiries for information have been ignored. But I’m going to re-establish a co-equal status.
I have a list of every inquiry the committee has made—many went unanswered by the Interior Department. We are going to send those inquiries again. We want answers to all of these questions. If we get the information, we proceed from there. If we are ignored again, we are in a realm where we need to use legal authority. We, as the majority, have options that we didn’t have as a minority. That includes subpoena.
HCN: How will you change the committee’s posture toward climate change and wildfire now that we’re seeing much longer fire seasons?
RG: There is no way to deny it. The planet is getting hotter, it is getting drier, and one of the things we need to do is confront the issue of these wildfires and other natural catastrophes that are happening—and confront them with the urgency that they demand. The president doesn’t think climate is the reason. That denial is embedded in Interior and this administration. We need to elevate the importance of addressing climate change.
Our response has been reactive. We have always dealt with the fire issue as a budgetary issue, and we should, but it’s more than that. The issue of climate change cannot be denied. We need to look at how we change building standards and where we decide it’s okay to build homes. In some cases, denial [of building permits] is a good thing.
HCN: How do you plan to handle the scandals and lawsuits facing Interior Secretary Ryan Zinke?
RG: There are 14 Inspector General investigations looking into Interior Secretary Ryan Zinke, and the Department of Justice is taking a look at him. (Editor’s note: So far, investigators have yet to confirm any wrongdoing.) At some point, those dogs start nipping at your heels. We’ve reached a point that Zinke needs to move on.
There’s no question that the extractive industries have had a free rein under Zinke. It’s no coincidence that Grand Staircase-Escalante National Monument was shrunk because of coal deposits. Bears Ears [National Monument] was shrunk because of nearby uranium deposits. The criteria of making decisions on how to best conserve and have a multi-use philosophy for our public lands and assets has been skewed in one direction and one direction only.
The extractive industries are protecting Zinke. He’s curried favor with them, and they are protecting him. He has a responsibility to protect the multi-use philosophy of the department. He’s failed. Resources are there not just for extraction.

domain: news


text:
Are you kidding? People should definitely not be offended when other people don’t want to read LGBTQ+ books. Why? Because everyone has their own preference. Everyone’s allowed to have their own preference.
It’s like dating. If someone has a preference in dating black guys is that racist to other races? Fuck no. Someone’s preference shouldn’t be an offence as long as that person’s preference isn’t based on bigotry and hate.
So untrue. My erotic male/male books are filled with hypersexual gay males and the readers love it. There certainly is nothing boring about it and in no way is it harmful. Gay males are proud of their sexuality and so tired of women who always have to shame them. Hypersexuality is a real aspect of gay male life, same as it is of straight male life. Those who don’t like it don’t have to read it. There are plenty of gay male books that depict them almost as nonsexual to appeal to those who don’t want to read a lot of sex.
Ignoring reality on a personal basis is fine, but suggesting that writing about realistic sex is ‘fetishizing’ is just not seeing life as it really is. Plenty of people, gay and straight, do all that stuff that those who push the ‘fetishize’ agenda don’t like, and they want to read about it too. To them it’s just reality, and good sex, and they have no need to pretend it’s anything else.
Sigh. You talk about not conflating personal feelings with reality, and yet here you are, talking past the point I was making because it is your own reality.
I am aware that sexuality is a different matter for many men in your community. It is in my posts.
That said, I was talking about cis women (often not even queer) who love to write about the gay experience and, in doing so, fetishize these men they are writing about to a point where it’s harmful to you and your peers.
You can be hypersexual all you want. Celebrate that with your fiction. Just don’t make every gay man out to be the cliché from a story and pressure people into it because it is true to your own experience. Not everyone is involved into the community the way you are, and it’s certainly none of us cis women’s business to stereotype you.
Because stereotyping is harmful, you know.
There are also plenty of men who don’t feel “gay enough” because of these images, so there’s that. Maybe consider other people sometimes.
Fun fact: One of my gay acquaintances claims that the series “Queer as Folk”, while highly criticized for its depiction of gays, is very accurate and that a first date is considered a failure when there is no sex involved. Make of that what you will.
Yes, that. Thanks for acknowledging that issue. <3
But I think in the end it comes down to this: every personal experience is valid, be it hypersexual (which is a logical way of celebrating your sexuality after being suppressed by society for a long time, or you just… like it xD) or more toned down because one is either not involved that much in the community and culture around all that or just… doesn’t care so much about sex in general.
I think it’s just important to be aware that neither experience is the standard. Maybe that’s where all of us can agree on.
I’d say asexual people have no business criticizing people who are sexual. They don’t feel it, they don’t know it, they have no idea. How can they tell sexual people how to be? It’s the same as heteros telling gays how to behave. It’s all a matter of not accepting others as they are. No one has the authority to say how many of anybody should behave this way or that for it to be right or wrong for them.
Better then this? nothing wrong with having style and topics you like to read LGBT is just a part of life…so are cars
would you get offended if some one didn’t want to read a car magazine…no
THIS. I would absolutely hate for all LGBTQ+ books to be thrown into one genre, because it would cut us off from the rest of the writing community. As a queer woman, I write a lot of queer characters, and I use tags to let people know (#lgbt#gay#lesbian#queer#bi etc. etc. etc.) It works just fine. If my book is a romance, a murder mystery, a science fiction work, a fantasy novel, whatever the case may be, I want it under that genre, not lumped under LGBTQ+. We want acceptance, right? So why would we resist being mainstreamed?
Pretty much how I feel, too. Sure, it doesn’t feel good when someone hates people that are like you, or when your story is turned down just because of LGBTQ+ characters or “themes,” but at the end of the day, it’s those people’s losses. Easier said than done, I know! But I don’t want homophobes reading my stories, anyway; I’m not writing my queer characters for them. I’m writing my stories for me and the other people who want to read them and see accurate queer representation Plus, we all know that fighting fire with fire doesn’t work. Better to leave hateful, fearful, angry people like that to their own devices, because honestly, that can’t be a fulfilling or happy life that they’re living with all that hate.
THIS. It’s like when people say that queer people or people of color shouldn’t be written into high fantasy novels. Like, what? So you’re okay with magic and dragons and giants and elves and fairies and unicorns, but not queer people or POC? Seems legit
OH MY GOD, THIS. I don’t even have anything to add because you’ve said it all. Just applause
Once again, no words, just applause Amen, sista!
Same. You don’t see a “Straight” genre, do you? And for good reason. We’re just like everyone else, don’t try to cordon us off from the rest of the stories on Wattpad.
Was wondering when people would bring up the fetishization of gay romances (cough “yaoi” “boys love” etc. etc.) by mainly straight women who think it’s “hot” As a queer woman myself, I actively avoid all stories that look like they fetishize queer people. Unfortunately, that’s so much of the queer fiction on Wattpad
Just going to chime in to say that I don’t really consider LGBT+ fic a genre, so much as a theme. Only exception might be a literal coming out story in which coming out and societal backlash is the main overarching focus of the story. Even then, it’d be a subgenre of realistic fic.
I think that most people who specify “no LGBT” are probably homophobic tbh. Someone who was actually concerned for how that makes LGBT people feel would probably say “no romance” or “no stories focused solely around LGBT themes” but that having LGBT characters was fine.
But, I don’t want to interact with whatever those people are doing anyway, tbh. Get them to read my story? And what, have their “criticism” be about how my character shouldn’t be trans? Not useful. Have them make a cover for me, and now something I’m using to identify my book was made by a homophobe? Also not really appealing to me. In a way I don’t really mind when people are upfront about not being cool with LGBT stuff because I know just to avoid them and not worry about it.
I hate when my stories get put into a reading list called yaoi. So many times I’ve wanted to be like can you change the name of the reading list or move my book into a different one. But I’ve always decided against it because it’s their life and not mine.
I personally have only been writing fanfictions or they start out as original books then I change them into that category because it’s less likely to come across homophobic people that way. Although, I do write in other genres every once in a while. But since I’ve been on here (almost five years now) I feel the fanfiction category is a little more friendly to the lgbt+ community (even though a good portion of people reading them are cis females fetishising gay males or trans people).
I agree with this paragraph, only, there are quite a few cases where no lbgt+ probably isn’t homophobic.
in my story, the MC is non-binary, so I imagined someone who specified no lgbt+ wouldn’t want to read it.
However, when looking for critiques, I found someone w/ that specification, but I gave them my story anyway just to see how they’d react (probably not the smartest idea but oh well) and they literally had no problem with it — (i was very confused)
in fact, they gave me a pretty positive review (they were probably just being nice but that’s not the point)
if I assumed no lgbt+ meant no lgbt+ characters or anything, I would’ve missed out on their feedback.
I honestly don’t know the correct way to respond to no lgbt+ comments b/c it could mean almost anything.
As I’ve said before, people need to specify what they mean by no lgbt+ because it can cause a lot of unnecessary hostility and confusion.

domain: sociology


text:
Activity of the epithelial Na+ channel (ENaC) is the limiting step for discretionary Na+reabsorption in the cortical collecting duct. Xenopus laeviskidney A6 cells were used to investigate the effects of cytosolic phospholipase A2 (cPLA2) activity on Na+ transport. Application of aristolochic acid, a cPLA2 inhibitor, to the apical membrane of monolayers produced a decrease in apical [3H]arachidonic acid (AA) release and led to an approximate twofold increase in transepithelial Na+ current. Increased current was abolished by the nonmetabolized AA analog 5,8,11,14-eicosatetraynoic acid (ETYA), suggesting that AA, rather than one of its metabolic products, affected current. In single channel studies, ETYA produced a decrease in ENaC open probability. This suggests that cPLA2 is tonically active in A6 cells and that the end effect of liberated AA at the apical membrane is to reduce Na+ transport via actions on ENaC. In contrast, aristolochic acid applied basolaterally inhibited current, and the effect was not reversed by ETYA. Basolateral application of the cyclooxygenase inhibitor ibuprofen also inhibited current. Both effects were reversed by prostaglandin E2(PGE2). This suggests that cPLA2 activity and free AA, which is metabolized to PGE2, are necessary to support transport. This study supports the fine-tuning of Na+ transport and reabsorption through the regulation of free AA and AA metabolism.
- sodium reabsorption
- arachidonic acid
- A6 cells
- cytosolic phospholipase A2
transport of sodium across principal cells in the distal collecting duct of the kidney is via an apical amiloride-sensitive epithelial Na+channel (ENaC) and a coupled basolateral Na+-K+-ATPase. This system vectorially moves Na+ from the external environment to the organism's internal environment. The rate-limiting step for this process is by regulation of ENaC. The principal means of regulating Na+transport in the cortical collecting duct is hormonal via the mineralocorticoid, aldosterone, and the peptide, antidiuretic hormone (19, 21, 40). Both hormones increase the apical membrane conductance to Na+, thereby promoting Na+reabsorption. Much effort has been focused on understanding the cellular mechanism of these hormonal regulatory events; however, the intracellular mechanism(s) responsible for their effects are not well identified. Less well described are the mechanisms of fine-tuning and local, intratubular, or cellular control of Na+ uptake.
Phospholipase A2, arachidonic acid, and ion channels.
The phospholipases A2 (PLA2) are a family of enzymes that metabolize membrane phospholipids and are generally classified into three groups, secretory (sPLA2), Ca2+ independent, and cytosolic PLA2(cPLA2) (33). Although the latter was originally termed cytosolic because it is found in the cytosol, this term is somewhat misleading because the active form of cPLA2 is membrane associated. cPLA2 has long been recognized to play an important role in a number of cellular processes. cPLA2 hydrolyzes the sn-2 position of phospholipids, resulting in the release of free fatty acids, primarily arachidonic acid (AA) in eukaryotic cells, and lysophospholipids. Both the free fatty acids and the lysophospholipids then may themselves mediate a number of responses by both intracellular and extracellular signaling. It is also well documented that a variety of plasma membrane receptors act on PLA2 (5, 33). Upon release, AA may initiate signaling or can be metabolized into a wide range of signaling factors by cyclooxygenases, lipoxygenases, and cytochromeP-450 monooxygenases, producing a cascade of prostaglandins, leukotrienes, and monooxygenase products, respectively. Determined by the predominate active metabolic pathways, the release of AA produces a prodigiously diverse range of physiological and pathological effects. In particular, many ion channels are affected by either AA itself and/or the various metabolites of AA (31). Free unsaturated fatty acids, principally AA, have varying effects on ion channels, thus a priori, the effects of AA on a given ion channel cannot be guessed. The effects of AA can either be inhibitory (23) or stimulatory (11, 13, 39). In addition, many studies involving the addition of free AA, usually at concentrations exceeding normal physiological levels, may not adequately address the physiological role of AA or its metabolites on ion transport.
In the 1970s, it was suggested that Na+reabsorption in the toad bladder was associated with an increased turnover of phospholipids (22), most likely through the involvement of PLA (42). To determine the possible effects of phospholipid turnover, and, in particular, AA on amiloride-sensitive Na+ channels and transepithelial Na+reabsorption, we chose to manipulate free AA levels by altering cPLA2 activity, using aristolochic acid as a relatively selective inhibitor of cytosolic PLA2, a strategy that had proved effective in previous work involving the maxi-K+channel in GH3 cells (13, 14).
This paper demonstrates that cPLA2 is tonically active in transporting A6 cells and that this activity is necessary for the support of transepithelial transport. Interestingly, AA is shown to reduce Na+ transport, whereas the metabolic product of AA, prostaglandin E2 (PGE2), is found to be necessary to maintain normal transport. Furthermore, the sidedness of cPLA2 inhibition suggests there is a local apical membrane-associated pool of cPLA2 at or near the apical Na+ channel in addition to the perinuclear and endoplasmic reticular localization of cPLA2.
MATERIALS AND METHODS
The 2F3 clonal line of A6 cells or A6 cells were grown on permeable supports under conditions that have been demonstrated to produce transepithelial Na+ transport, as well as a predominance of the low-conductance highly selective amiloride-sensitive Na+ channel in the apical membrane (21). Open-circuit current measurements were made using the 2F3 clonal cell line (which, in general, produces more current than the A6 cell line). Short-circuit current and single channel measurements were made using the A6 cell line. It should be noted that A6 cells were found to respond identically to the 2F3 cell line in open-circuit current measurements, albeit with a lower magnitude of current. Cells were maintained at 26°C in 4% CO2 in a mixture of 7:3 Coon's F-12 and Leibovitz's L-15 with 25 mM NaHCO3 (pH 7.4), 10% fetal calf serum, 1% streptomycin, and 0.6% penicillin. Cells for transepithelial current measurements and assays involving free AA production or cPLA2 activity were grown on 25-mm Anapore supports (Nunc). Cells for patch-clamp analysis were plated on rat tail collagen-coated Millipore-CM filters attached to the bottom of custom-made Lucite rings. Cells on both types of permeable supports were grown to confluency in the above conditions in the presence of 1.5 μM aldosterone. Under these conditions, high-resistance monolayers formed within 7–14 days. Short-circuit current measurements were performed on A6 cells grown under the conditions reported by Blazer-Yost et al. (4).
Measurement of transepithelial current.
The quality of high-resistance monolayer formation was monitored using an epithelial voltohmmeter (EVOM; World Precision Instruments). This instrument consists of a pair of Ag-AgCl electrodes mounted in “chopstick” fashion attached to a customized voltohmmeter. Both transepithelial potential (in mV) and transepithelial resistance (in KΩ) were measured with this instrument. Transepithelial current was calculated by Ohm's law, expressed as μA/cm2, and is referred to as open-circuit current in the text to clearly distinguish it from the short-circuit currents measured in Ussing chambers. Although the open-circuit current technique tends to underestimate the amount of current that would be obtained in short-circuit current measurements, EVOM measurements proved consistent and reliable for detecting changes in transepithelial voltage, resistance, and current. Because cells used in EVOM measurements were at room temperature and not aerated with 95:5 gas, current tended to slowly decline with time (see Figs. 1 A and 7 A). Experiments were performed in serum-free A6 culture media. Upon replacement of serum- containing media with serum-free media, currents typically become relatively stable within 5 min. To corroborate open-circuit current measurements, conventional short-circuit current measurements were performed. A6 cells were grown to confluency on Transwell supports, and the supporting membrane was then placed in an Ussing chamber for measurement of short-circuit current as described by Blazer-Yost et al. (3). Current from individual monolayers was allowed to stabilize (∼1–2 h) before drug addition. Data were recorded on a strip chart recorder, and current magnitudes were subsequently measured for plotting. No qualitative difference was observed between the methods of measurement or the cell line used (2F3 vs. A6).
A commonly used indirect approach to assess the release of free AA involves monitoring the release of [3H]AA from cells (1). A6 cells on permeable supports were loaded with [3H]AA for 24 h. During this time, the [3H]AA was incorporated into the cellular lipids. Cells were then washed four times in PBS, and the efflux of labeled AA was monitored by a standard sample-and-replace method using fatty acid-free BSA in the efflux media to act as a trap for liberated free fatty acid. Samples were placed in scintillation cocktail (BioSafe II; Amersham), and radioactive decay was counted using a Packard 1900A scintillation counter. At the end of the sample period, cells were lysed in 0.1 N NaOH, and the cell lysate was counted to determine the counts remaining in the cells. Data were plotted as the percent counts remaining in the cells for each time point, and the slope of the relation was used to determine the relative rate of free AA release. This technique offers an advantage over PLA2 activity measurements in cell lysates in that efflux (AA release) can be monitored on the apical side of the monolayer, thus providing a more accurate assessment of the free AA at the apical membrane.
Single channel patch-clamp recording.
Cell-attached single channel patch data of ENaC were obtained as previously described by Kemendy et al. (25) and Ma and Ling (29). Briefly, A6 cells were cultured on custom Lucite rings and viewed under Hoffman modulation optics mounted on a Nikon Diaphot inverted microscope. Patch pipettes with a tip resistance of 7–9 MΩ were fabricated from TW150 glass on a Narishige PP-83 puller and fire-polished with a Narishige MF-83 polisher. Bath and pipette solutions contained (in mM): 96 NaCl, 3.4 KCl, 0.8 CaCl2, 0.8 MgCl2, and 10 HEPES at pH 7.4 (titrated with 1 N NaOH). Single channel currents from apical membrane patches were recorded with an Axon 1D amplifier and data digitized to a computer using a TL-1 data interface. Single channel data analysis was accomplished using pCLAMP software (Axon).
[3H]AA was obtained from Amersham. Aristolochic acid, ibuprofen, 5,8,11,14-eicosatetraynoic acid (ETYA), 7,7-dimethyleicosadienoic acid (DEDA), and PGE2were obtained from Biomol. All other reagents were obtained from either GIBCO BRL or Sigma.
Data are presented as means ± SE. Significance was estimated using the Student's t-test with P < 0.05 indicating a significant difference.
Inhibition of cPLA2 by application of the cPLA2 inhibitor, aristolochic acid, produced opposite effects depending on whether application was to the apical or basolateral side of the transporting epithelial monolayer. We show (below) that inhibition of cPLA2 from the apical side produces an increase in transepithelial current, whereas inhibition from the basolateral side produces an inhibition of transepithelial current. The apical effect is due to the downmodulation of apical Na+ channels (ENaC) by free AA produced by cPLA2 activity. Current inhibition after inhibition of cPLA2 from the basolateral side results predominately from an inhibition of PGE2 production within the cells. It is important to note that both results imply that cPLA2 is present and active in the aldosterone-stimulated monolayers and that disruption of cPLA2 activity has significant effects on the magnitude of transepithelial Na+ transport.
Inhibition of cPLA2 from the apical side.
The results of cPLA2 inhibition after apical application of aristolochic acid is shown in Fig.1. Open-circuit transepithelial current shows a marked increase on application of aristolochic acid to the apical surface. Current was 6.8 ± 0.2 μA/cm2 in the control monolayers and 7.5 ± 0.3 μA/cm2 in the treated group at time zero. With the addition of an apical media containing 200 μM aristolochic acid, current increased significantly within 1 min to 13.6 ± 0.6 μA/cm2(P < 0.001; Fig. 1 A). Addition of replacement apical media to the control group resulted in no significant change in current. Currents remained elevated approximately twofold for at least 5 min, with control and aristolochic acid-treated groups having currents of 5.6 ± 0.2 and 13.6 ± 0.5 μA/cm2, respectively. Apical aristolochic acid reduced transepithelial resistance from 1.22 ± 0.09 KΩ in control to 0.95 ± 0.06 KΩ. The increase in current with apical aristolochic acid was fully reversible, as shown in Fig. 1 B. Open-circuit transepithelial current was 5.9 ± 0.2 μA/cm2 before the apical addition of aristolochic acid. Apical aristolochic acid produced an increase in current to 9.6 ± 0.3 μA/cm2. Upon washout of apical aristolochic acid, currents were not significantly different from control, 5.9 ± 0.3 μA/cm2. All current was amiloride sensitive (data not shown).
In an effort to more fully describe the nature of current stimulation seen in the open-circuit current measurements with apical application of aristolochic acid, the short-circuit current method was applied. Identical results to those obtained in open-circuit conditions were observed upon apical application of 200 μM aristolochic acid under short-circuit conditions (Fig. 2). Starting current was 22.7 ± 1.4 μA/cm2 for the control group of monolayers and 20.3 ± 2 μA/cm2 for the treatment group. Current did not vary significantly in the control group over the course of the experiment, with the ending current being 22.7 ± 1.4 μA/cm2. Significant current stimulation was seen with the apical application of aristolochic acid within 20 s of addition (30.8 ± 1.6 μA/cm2,P < 0.001 vs. current at time 0). Current remained elevated during the 3-min time course with current at 40.2 ± 2.9 μA/cm2. The addition of 1 × 10−5 M amiloride to the apical bathing media reduced current in both the control and the aristolochic acid-treated groups, with current being 3.3 ± 1.1 and 3.5 ± 1.3 μA/cm2, respectively. There was no significant difference in the amiloride-insensitive current component between each group.
Because there have been some reports that aristolochic acid can affect the activity of sPLA2 (41), a specific inhibitor of sPLA2 was applied to the apical side of transporting monolayers. Figure 3 clearly demonstrates that 10 μM DEDA does not affect open-circuit transepithelial current. Current level began at 7.1 ± 0.2 μA/cm2, and addition of DEDA to the apical side did not significantly increase current; indeed, current was slightly decreased to 6.4 ± 0.3 μA/cm2. However, subsequent application of 100 μM apical aristolochic acid produced a significant increase in transepithelial current (12.5 ± 0.3 μA/cm2, P < 0.001). Thus the apical aristolochic acid effect is not due to actions on sPLA2.
Presumably, inhibition of cPLA2 by apical aristolochic acid application should reduce the amount of free AA present at the apical membrane. One means of accessing the liberation of free AA is to measure the release (efflux) of labeled AA from the monolayers. Figure4 A shows the efflux of [3H]AA from monolayers that had [3H]AA incorporated into the cellular lipid pool. Apical application of 200 μM aristolochic acid, as presumed, resulted in a decrease in the release (efflux) of free AA from the apical surface of the transporting monolayers. Although small, the efflux rate of [3H]AA from the apical surface was significantly reduced with apical aristolochic acid application (Fig. 4 B). Efflux rates were 0.045 ± 0.013 s−1 for the untreated group and 0.019 ± 0.004 s−1 for monolayers treated with apical aristolochic acid (P < 0.05). It should be mentioned that the actual release of free AA is underestimated by monitoring the efflux of AA, because free AA will partition into the cellular membrane and the cytosol as well as the external media. Nonetheless, a decrease in AA release was observed, thus indicating that available free AA at the apical membrane is reduced by the apical application of aristolochic acid. No appreciable change in [3H]AA efflux was detected with apical application of DEDA or with the basolateral application of aristolochic acid (data not shown).
The cPLA2 liberation of free AA from the sn-2 position of membrane lipids results in the production of a lysophospholipid product. Either free AA or the lysophospholipid product may act as a signaling molecule, and, in addition, free AA is the substrate for a number of enzymes and can result in a prodigious number of different signaling molecules. In an effort to ascertain which signaling pathways are involved, an analog of AA containing four triple bonds (ETYA) vs. the four double bonds of AA can be employed. ETYA acts as an unsaturated free fatty acid as well as an inhibitor of the enzymes that normally metabolize free AA. It has thus been used to distinguish between free fatty acid effects and AA metabolic product effects. As shown in Fig. 5, a 2-min apical application of 40 μM ETYA fully reverses the effect of current stimulation with apical application of aristolochic acid, thus implying that free AA is responsible for decreasing transepithelial current. Control currents were 5.7 ± 0.4 μA/cm2, which increased to 9.3 ± 0.7 μA/cm2 with apical application of 200 μM aristolochic acid. Subsequent apical addition of ETYA reduced currents to control values (5.6 ± 0.4 μA/cm2) and increased transepithelial resistance from 0.99 ± 0.05 KΩ to 1.16 ± 0.06 KΩ. Apical application of ETYA on control monolayers resulted in an ∼9% reduction of basal current, from 4.3 ± 0.2 to 3.9 ± 0.2 μA/cm2, which did not represent a significant difference (n = 12, data not shown). The lack of significance may arise from the presence of endogenous AA masking the effect of ETYA in the basal state.
To determine whether the stimulation of transepithelial current observed with apical application of aristolochic acid in monolayers was due to a free fatty acid-induced reduction in ENaC activity, the patch-clamp technique was employed. Figure6 shows the results of cell-attached patch-clamp recordings on A6 cells, specifically ENaC activity. Typical single channel records are shown in Fig. 6 A. After addition of 40 μM ETYA, the single channel open time decreases. With extensive washout, this decrease in channel open time can be partially reversed. A slight decrease in channel amplitude is also observed; however, this is not due to a change in the single channel conductance, as shown in Fig. 6 B, but rather represents a shift in the cell membrane potential, thus affecting the patch-clamp potential (∼8 mV). The voltage shift is not sufficient to produce the profound effects on channel open time observed in Fig. 6 A. Indeed, channel open time in the presence of ETYA is also decreased from control values if the patch-clamp potential is adjusted such that the channel amplitude remains constant (e.g., a constant patch potential, data not shown). Figure 6 C shows the mean results obtained from eight patches. ENaC open probability (P o) was 0.43 ± 0.05 before ETYA and was reduced 56 ± 9% to 0.18 ± 0.04 (P < 0.002) with the application of ETYA. Subsequent washout led to a partial reversal with a mean channelP o of 0.30 ± 0.03%.
Inhibition of cPLA2 from the basolateral side.
Unlike the results of cPLA2 inhibition from the apical side of the monolayer, basolateral application of aristolochic acid produced a decrease in transepithelial current.1 Figure7 A shows the results of basolateral application of 200 μM aristolochic acid on open-circuit current. Relative current is plotted. Starting current amplitude was 5.2 ± 0.09 μA/cm2 for both the control and treatment groups. The replacement of basolateral media results in a slow rundown of transepithelial current, with current being reduced to 3.1 ± 0.1 μA/cm2 after 20 min. The inclusion of aristolochic acid in the basolateral media results in a significant decrease (over control) in transepithelial current. The decrease over control is first noted and significant (P < 0.05) at ∼3 min and continues to develop for the 20-min time course presented. The time course is thus slower than that of the apical effect shown in Fig. 1 A that was complete at 1 min. Currents at the 5-min time point were 4.7 ± 0.1 μA/cm2 for control and 3.7 ± 0.1 μA/cm2 for basolateral aristolochic acid. This difference was greater at the 20-min time point where currents were 3.1 ± 0.1 and 1.3 ± 0.08 μA/cm2 for control and basolateral aristolochic acid-treated groups, respectively. Transepithelial resistance rose from 1.29 ± 0.07 KΩ control to 1.56 ± 0.08 KΩ at the 15-min time point.
Inhibition of transepithelial open-circuit current by basolateral application of aristolochic acid was not reversed by the basolateral application of ETYA. This differs from the effect of ETYA on the increase in current observed with apical application of aristolochic acid (see Fig. 5). Figure 7 B shows the results when ETYA is added to the basolateral media subsequent to basolateral aristolochic acid application. Control open-circuit current was 3.8 ± 0.2 μA/cm2. Current was reduced to 1.8 ± 0.1 μA/cm2 with a 10-min basolateral application of aristolochic acid. Subsequently, a 2-min application of basolateral ETYA (40 μM) significantly reduced currents further to 1.0 ± 0.06 μA/cm2 (P < 0.01). This indicates that the inhibition of current seen with basolateral application of aristolochic acid is not due to free fatty acid and implies that a metabolic product of AA may be involved in maintaining transepithelial transport.2
The effect of basolateral application of aristolochic acid was also measured under short-circuit current conditions and is shown in Fig.8. The results did not differ from those observed in the open-circuit conditions (Fig. 7 A). Current began at 27 ± 6 μA/cm2 for both control and experimental groups. Unlike the open-circuit current conditions, current level under short-circuit current was maintained throughout the experiment for the control group (26 ± 6 μA/cm2 at 10 min). Addition of basolateral aristolochic acid (200 μM) resulted in a marked decrease in short-circuit current, with a significant decrease clearly observed at 2 min (P < 0.05) and a large decrease observed at 10 min, 7 ± 2 μA/cm2with aristolochic acid vs. 26 ± 6 μA/cm2 for the control group. Addition of 1 × 10−5 M amiloride reduced currents to similar levels of 4.2 ± 0.9 and 4.1 ± 0.9 μA/cm2 for control and experimental groups, respectively.
Perhaps the most probable AA metabolite to mediate support of transepithelial transport is the cyclooxygenase product of AA, PGE2. Chronic application of basolateral PGE2has been reported to increase Na+ channel number in the apical membrane of frog skin (17) and in A6 cells (27, 30). If PGE2 was necessary for transport, then inhibition of cPLA2 might produce an inhibition of transport as observed in Figs. 7 and 8, and the current inhibition should be prevented by the addition of exogenous PGE2 to the basolateral side of the monolayer. Such experiments are summarized in Fig. 9. Open-circuit current was measured under three conditions: basolateral aristolochic acid alone, basolateral PGE2 alone, and simultaneous application of basolateral aristolochic acid and PGE2. Starting currents were 3.2 ± 1.2 μA/cm2 for each group. As shown previously, basolateral application of aristolochic acid led to a significant reduction in transepithelial current (1.6 ± 0.2 μA/cm2 at 15 min). The basolateral application of 10 μM PGE2 alone resulted in the stimulation of transepithelial current (5.8 ± 0.2 μA/cm2 at 5 min). The addition of basolateral PGE2 with basolateral aristolochic acid prevented the basolateral inhibition of transepithelial current caused by basolateral aristolochic acid application alone. Current levels in the presence of both basolateral PGE2 and aristolochic acid were 4.8 ± 0.3 at 5 min and 6.7 ± 0.5 μA/cm2at 15 min.
Direct inhibition of cyclooxygenase by basolateral ibuprofen leads to an inhibition of transepithelial open-circuit current similar to that seen with cPLA2 inhibition by basolateral aristolochic acid (Fig. 9 B). Starting currents were 5.2 ± 0.2 μA/cm2 for each group. Transepithelial current was reduced to 3.5 ± 0.2 and 2.9 ± 0.2 μA/cm2with basolateral application of aristolochic acid or ibuprofen, respectively. Current reduction in both cases was reversed with basolateral addition of 10 μM PGE2. Current 5 min after basolateral PGE2 was 4.8 ± 0.3 μA/cm2in monolayers treated with basolateral aristolochic acid and 6.6 ± 0.4 μA/cm2 for monolayers treated with basolateral ibuprofen. The difference in current magnitude after PGE2application is interesting and is addressed in discussion.
Combined effect of cPLA2 inhibition from the apical side and basolateral PGE2 addition.
The increase in transepithelial current observed with the apical application of aristolochic acid is additive, with the stimulation of current observed with the basolateral application of PGE2. Figure 10 shows transepithelial open-circuit current measurements from monolayers treated simultaneously with apical aristolochic acid (200 μM) and basolateral PGE2 (10 μM). Starting current was 8.9 ± 0.3 μA/cm2. Combined application of apical aristolochic acid and basolateral PGE2 resulted in a peak increase in current of 21.2 ± 1.3 μA/cm2 at 10 min. Peak current magnitudes observed with apical application of aristolochic acid alone (Fig. 1 A) were 14.2 ± 0.6 μA/cm2, whereas those observed with basolateral PGE2 were 6.7 ± 0.5 μA/cm2 (Fig. 9, A and B). The sum of these current magnitudes, 20.9 μA/cm2, is not significantly different from the 21.2 ± 1.3 μA/cm2observed in Fig. 10.
The data presented demonstrate that cPLA2 is tonically active in transporting A6 cells and that this activity is necessary for the support of transepithelial transport. Interestingly, however, a sidedness to cPLA2 inhibition results in a divergent effect on transepithelial transport, with inhibition from the apical side resulting in an increase in current and inhibition from the basolateral side resulting in a decrease in current. The increase in transepithelial current observed with apical side inhibition of cPLA2 results from the downmodulation of ENaC by free AA. The decrease in transepithelial current observed with basolateral side inhibition of cPLA2 results in part from a decrease in the metabolic product of AA, PGE2. Furthermore, the sidedness of cPLA2 inhibition suggests there is a local apical membrane-associated pool of cPLA2 at or near the apical Na+ channel in addition to commonly found perinuclear and endoplasmic reticular localization of cPLA2. These data provide a novel and intriguing mechanism whereby transepithelial Na+ transport and thus fluid reabsorption can be modulated over a relatively rapid time course, as well as provide for differential modulation by apical or basolateral signaling mechanisms.
The apical effect.
Free AA or free unsaturated fatty acids have been shown to affect a number of ion channels (31), producing either an increase (11, 12, 39) or a decrease (23) in single channel P o. The data presented herein not only demonstrates that free fatty acid reduces the P oof the apical Na+ channel, ENaC, but also shows that under normal conditions of transport (e.g., aldosterone stimulated), free AA is produced at the apical membrane and exerts a downmodulatory effect on ENaC, thus slowing transport. Inhibition of cPLA2, by aristolochic acid from the apical side of transporting A6 cell monolayers, produced an approximate twofold increase in transepithelial Na+ transport and a 58% decrease in the rate of AA liberated at the apical membrane. Apical ETYA reversed the stimulatory effect of apical aristolochic acid on transepithelial current, indicating that free AA itself was acting to downmodulate transport at the apical membrane. Although apical AA application produced similar results (not shown), these results were more variable and could be misleading due to cellular metabolism of AA into a variety of signaling molecules. ETYA provided an advantage in that it is an analog of AA and has similar effects on ion channels as AA (12); however, ETYA is not subject to cellular metabolism. Apical ETYA might also inhibit PGE2 production, resulting in a reduction of current as with the basolateral effect of aristolochic acid or ibuprofen. However, if this were the principal means of the apical ETYA reduction of current, one would not expect current inhibition by AA or an additive effect of basolateral PGE2 and apical aristolochic acid as is observed. In single channel recordings, ETYA reduced the P o of ENaC. The reduction in ENaC activity with ETYA supports the notion that tonic production of free AA at or near the apical membrane downmodulates ENaC activity and thus transepithelial transport of Na+. The fact that an increase in transepithelial transport is observed with the inhibition of cPLA2 from the apical side supports the presence of tonically active cPLA2 at or near the apical Na+ channel in the transporting A6 cells. Alternatively, AA liberated at a locale distant to ENaC within the cell might be responsible for downmodulation of ENaC activity. This alternative hypothesis is less likely because inhibition of cPLA2 by basolateral application of aristolochic acid does not produce the same affect as apical application.
The basolateral effect.
Inhibition of cPLA2 from the basolateral side of the A6 cell monolayer produced an effect on transepithelial current that was opposite from inhibition from the apical side. Basolateral application of aristolochic acid produced an inhibition of transepithelial current with a time course somewhat slower than that which was observed for the increase in current with apical aristolochic acid. Unlike that of the apical side effect, ETYA did not relieve the inhibition of current seen with basolateral side inhibition of cPLA2 but rather led to a greater inhibition of current. This effect of ETYA is consistent with the notion that free AA is tonically produced within the cell and is being metabolized to products that act to support transepithelial transport. Basolateral addition of the cyclooxygenase product of AA metabolism, PGE2, was able to prevent as well as reverse the inhibitory effect of basolateral aristolochic acid application. In support of PGE2 involvement, basolateral application of the cyclooxygenase inhibitor, ibuprofen, produced transepithelial current inhibition similar to that of aristolochic acid. The effect of ibuprofen was also reversed by the addition of PGE2. Interestingly, PGE2 produced a more robust response subsequent to ibuprofen than subsequent to aristolochic acid. Inhibition of cPLA2 would result in a decrease in all metabolic products of AA, whereas cyclooxygenase inhibition would only decrease prostaglandin production. Thus the more robust response with PGE2 after ibuprofen may indicate the involvement of other AA metabolites involved in the support of transport. Indeed, the lipoxygenase product of AA, LTD4, has been shown to increase Na+ channel activity in A6 cells (8).
The inhibitory effect on transport by basolateral side inhibition of cPLA2 supports the notion that cPLA2 within transporting A6 cells is active, and along with cyclooxygenase activity, is necessary to support Na+ transport. Indeed, basolateral PGE2 has previously been shown to increase short-circuit current and Na+ conductance in A6 cells (27, 30) and frog skin (17).
Model for the divergent effect.
The divergent effect on transepithelial current with inhibition of cPLA2 from the apical and basolateral sides suggests discrete intracellular pools of cPLA2. Although cPLA2 has predominately been localized to the perinuclear and endoplasmic reticular membranes within cells (35), several studies have reported evidence that suggests that cPLA2 also can occur at the plasma membrane in confluent bovine endothelial cells (36), in fibroblasts (7), in glomerular epithelial cells (28), and in bradykinin-stimulated Madin-Darby canine kidney cells (26). Perhaps the most convincing evidence for cPLA2 presence in the plasma membrane derives from inside-out patch-clamp experiments in GH3 cells, where functionally, cPLA2 occurs in the same membrane patch as the Ca2+-activated K+ channel (BK channel) (14). Ion channels occur in the plasma membrane at relatively low abundance. At minimum, if one cPLA2 was associated with one BK channel, one might expect the plasma membrane abundance of cPLA2 to also be relatively low, particularly compared with the nuclear envelope.
A receptor-coupled apical membrane-situated cPLA2 could act as the transduction mechanism for intraluminal signaling molecules. The stimulation of Cl− current in colonic epithelia by 5′-(N-ethylcarboxamido)adenosine (an adenosine analog) was found to correlate with the release of [3H]AA and was abolished by the PLA2 inhibitor 4-bromophenacyl bromide, suggesting that adenosine was acting by increasing PLA2activity, and the resulting free AA was affecting transepithelial Cl− current (1). Similarly, Smallridge and Gist (38) have shown that the ATP stimulation of125I− efflux in the thyroid cell line FRTL-5 was correlated with an increased release of [3H]AA, both of which were abolished by the PLA2 inhibitor U-73122. It is interesting to think of these results in light of the connection between ATP, Cl− secretion, and Na+reabsorption, where it is speculated that external ATP, somehow dependent on active cystic fibrosis transmembrane conductance regulator (CFTR) Cl− channel, acts to inhibit Na+reabsorption and the Na+ channel (15). Endothelin-1 has also been shown to increase free AA levels (24) in smooth muscle and to affect Na+transport in epithelia (20). The emerging precedence from these data suggests that receptor-mediated signaling through cPLA2 and AA release can have an effect on ion transport processes. Receptors to adenosine, ATP, and endothelin-1 have been identified in renal collecting duct epithelia either directly (10, 16, 34, 37) or by inference from pharmacological effects (see below). Moreover, as might be expected for intratubular signaling, adenosine, ATP, and endothelin-1 are present in normal urine and have been shown to vary during certain pathological conditions (24, 32). Their occurrence, particularly in the cases of adenosine and endothelin-1, are proposed to protect against ischemic damage by inhibiting the transport of Na+(2, 6, 18). As briefly mentioned earlier, the amiloride-sensitive Na+ channel is inhibited by extracellular ATP, the level of which is partially determined by CFTR activity (15), thus allowing for cellular self-regulation of apical Na+ reabsorption. Additionally, apical application of adenosine in A6 cells, via a type A2receptor, produces an inhibition of Na+ channel activity (29). Basolateral application of adenosine, on the other hand, produces an increase in Na+ transport (9). Endothelin-1 applied basolaterally, most likely ETA receptor mediated, also produces a stimulation of apical Na+ channel activity (20), thus suggesting a precedence that compounds, which may act through cPLA2, produce opposite effects when applied to the apical membrane vs. the basolateral membrane. Studies that determine the effects and dependence of apical agonists on cPLA2activity, free AA release at the apical membrane, and Na+channel activity are currently under way.
On the basis of the data presented, we propose that 1) AA from cPLA2 activity at or near the apical membrane is primarily responsible for downmodulation of transepithelial current, and 2) AA from cPLA2 activity combined with cyclooxygenase activity within the cell, presumably perinuclear, produces PGE2, which is necessary to support transport. Together, the apical effect of AA downmodulation of ENaC and the necessity of the AA product PGE2 to support transport provide a cellular signaling mechanism to fine-tune Na+transport, based on the relative activity of cPLA2 at the apical membrane and the perinuclear cPLA2 and cyclooxygenase activity. With fixed cPLA2 activity, manipulation of cyclooxygenase activity would result in relative changes of free AA. With greater cyclooxygenase activity, more PGE2 would be produced, resulting in less free AA, which would tend to maximize transport. In contrast, less cyclooxygenase activity would yield less PGE2 and result in more free AA, which would tend to minimize transport. Increased cPLA2activity at or near the apical membrane would tend to decrease transport, whereas increased cPLA2 near an active cyclooxygenase enzyme would tend to increase transport. Indeed, as early as 1975, an increased phospholipid turnover was suggested to occur with stimulation of Na+ transport in toad bladder (22). This increase was attributed to cellular increase in phospholipase activity (42). This report provides further evidence as well as a model for the increased rate of phospholipid turnover and establishes the increased turnover as important in the regulation of Na+ transport. Specifically, AA and PGE2 act in an antagonistic manner in A6 cells to modulate Na+ transport rate.
We are grateful to B. J. Duke for plating and maintaining the 2F3 cell cultures. All short-circuit current experiments were made in the laboratory of Bonnie Blazer-Yost (Dept. of Biology, Indiana Univ. Purdue Univ. at Indianapolis). We are most appreciative of her support in providing equipment and the expertise of her laboratory personnel, Carla J. Faletti, Amy Hartman, and Diane Hoover. Without this support, the short-circuit current measurements would have been much more difficult.
This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Grant DK-09215 (to R. T. Worrell), National Science Foundation Grant IBN-9603837 (to D. D. Denson), NIDDK Grants DK-37963 and DK-50268 (to D. C. Eaton), and by the Center for Cell and Molecular Signaling at Emory.
Address for reprint requests and other correspondence: R. T. Worrell, Dept. of Physiology, Center for Cell and Molecular Signaling, Emory Univ., Atlanta, Georgia 30322 (E-mail:).
↵1 It is interesting to note that long-term (>15–20 min) application of aristolochic acid to the apical side also resulted in an inhibition of transepithelial current subsequent to current stimulation. The most likely explanation is that aristolochic acid is diffusing through or across the cell and reaching the sites that are more readily accessible from the basolateral membrane (the basolateral effect but with a lag time to the onset). All apical application experiments were performed with <10 min of aristolochic application to avoid this secondary effect. Transepithelial current increase was never observed subsequent to the decrease in current with basolateral aristolochic acid, thus suggesting a necessity for cPLA2 activity in the support of transport.
↵2 Experiments with basolateral ETYA alone also produced inhibition of current similar to basolateral aristolochic acid. This arises from ETYA inhibition of the enzymes that metabolize AA (i.e., cyclooxygenase). ETYA was used in this experiment as a means of distinguishing between a free fatty acid effect vs. an AA metabolite effect.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2001 the American Physiological Society

domain: chemistry


With the rapid advance of mobile computing technology, there is a significant demand for mobile application testing on real devices. In mobile app testing, Mobile Device Management is playing a vital role and understanding the challenges in Mobile Device Management is as important as solving them. In order to avoid Device specific problems, test automation developers must test their apps on a large number of the devices, which is costly and inefficient. In this talk, we demonstrate how to leverage machine learning algorithms in mobile test automation. Participants will learn how to use ML algorithms in mobile device selection, Supervised learning and Prediction using a couple of algorithms.


text:
FHA borrowers have to pay two types of mortgage insurance premiums: annual and upfront. The upfront mortgage insurance premium is charged when you first get your mortgage, and the annual premium is an ongoing obligation you pay every year. Paying for FHA mortgage insurance. The upfront mortgage insurance premium costs 1.75% of your loan amount.
FHA Mortgage insurance. fha mortgage insurance varies from 0.45% to 1.05% of the loan amount. It usually remains for the life of the loan.
FHA Loans 101: You Might Actually Be Able to Buy a House – If you’re shopping for a new home, you might come across something called an FHA loan. Named for the federal housing administration, the agency that insures them, these loans are designed to make.
FHA Loan Requirements Important FHA Guidelines for Borrowers. The FHA, or Federal Housing Administration, provides mortgage insurance on loans made by fha-approved lenders. fha insures these loans on single family and multi-family homes in the United States and its territories.
How FHA Loans Work You get an FHA loan from an FHA-approved mortgage lender. The loan is insured by the Federal Housing Administration. Because of that insurance, the credit and income requirements for an FHA loan are more lenient.
FHA loans are government-insured mortgages with less-rigorous criteria for borrowers. Learn how this loan could be right for you with our 2019.
fha home loan Credit Score Credit Scores For An FHA Home Loan: When Low Credit Is Not. – FHA home loan rules specify that the lender must insure a borrower as a 500 FICO score at the very minimum in order to qualify for an FHA mortgage, but your credit score numbers don’t tell the whole story.
FHA Loans – FHA Property Appraisal – FHA-Home-Loans.com FHA loan info from a fha mortgage loan site specialized in FHA loans – FHA Home Loans.com is not a Government Agency.
Apply for FHA home loan in Texas, New Mexico & Arizona with First. The U.S. Department of Housing and Urban Development (HUD) insures these loans.
Federal Housing Loan Requirements First Time Home Loans | CalHFA FHA Program – CA.gov – How do I apply for this loan program? Since CalHFA is not a direct lender, our mortgage products are offered through private loan.
The Nation’s Housing: FHA changes crack down on higher-risk mortgages – Here’s what’s happening: For several years, FHA has insured loans to buyers who previously would have been considered too risky or marginal at best. Those applicants often carried crushing monthly.
Mortgages | USAGov – Learn some of the basics about mortgages. The .gov means it’s official. Federal government websites often end in .gov or .mil.
HUD.gov / U.S. Department of Housing and Urban Development. – The Federal Housing Administration, generally known as "FHA", provides mortgage insurance on loans made by FHA-approved lenders throughout the United States and its territories. FHA insures mortgages on single family and multifamily homes including manufactured homes and hospitals.

domain: economics


Scoot Over Bodysuits
Brand: Greendigo Organic Clothing
Our super soft organic cotton Bodysuits in light grey features a bright orange scooter detail for a playful touch. The all-along side snap kimono opening makes it easier to put on and change nappies. Can be worn by itself or as a vest.
More on the Design: Soft and breathable | Full sleeved | Snap button opening | Applique detail | No itchy labels | Environmentally friendly non-toxic colors | Machine washable | Certified organic by Global Organic Textile Standard (GOTS) | Made in India
Fabric & Care: 100% GOTS certified organic cotton. Non-toxic dyes and colors, suitable for kids’ skin. Machine wash at 30 degrees (A higher temperature wash will not harm the clothes but is not eco-friendly). Do not bleach. Warm Iron. Dry in Shade.


Two Olympians, a former governor and a rapper-turned-contractor will try to dance their way into America’s hearts.
The cast of Season 23 of “Dancing With the Stars” was revealed Tuesday morning on “Good Morning America.”
Vying for the Mirror Ball trophy this time around are:
Olympic swimmer Ryan Lochte Olympic gymnast Laurie Hernandez Former “Brady Bunch” star Maureen McCormick Talk show host Amber Rose “Little Women: LA” reality star Terra Jole Former “Taxi” star Marilu Henner Former Texas Gov. Rick Perry Former child star Jake T. Austin Former pro football player Calvin Johnson Super producer and recording artist Kenny “Babyface” Edmonds Former rapper Vanilla Ice Race car driver James Hinchcliffe Country singer Jana Kramer
“I think it’s time to hang up my Speedos and put on my dancing shoes,” Lochte said Tuesday morning on “GMA.” “I’m going to treat this kind of like going into the Olympics.”
The new gig comes at a good time for Lochte, who returned from the Olympics under a dark cloud over lying about being robbed at gunpoint in Rio. Four major sponsors walked away from him.
This season will also welcome back some veterans.
Derek Hough confirmed on Instagram Monday that he would be returning to the dance floor.
Hough took some time off after winning Season 21 with Bindi Irwin. He had been set to star this fall in the Broadway revival of “Singin’ in the Rain.” But he explained that the production had been unable to secure a theater and has been postponed.
“It’s always great to be back with my DWTS family, coaching, creating and making lasting memories,” he wrote.
Two-time champion Cheryl Burke is also returning and will partner with Lochte. She last competed with partner Antonio Sabato Jr. during Season 19 in 2014.
“For me it was about performing again,” she said following the announcement. “I am ready to be back and do what I was born to do.”
“Dancing With the Stars” returns September 12 on ABC.