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Not great for teens
Sail Date: March 15, 2015
Reviewed: 2 years ago
Traveled As: Family (older children)
Room Type: Inside
Our interior cabin was way to small for four people and the ship didn't really have much for my two teens to do on the days at sea. The freestyle dining to us meant wait in line for 30 minutes and then wait with a pager for 30 minutes. Two positives...food was great once you got it and shows were great.
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|Ship Quality
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|Food and Dining
|but had to wait to get it every night but one
|Service and Staff
|Had to wait every night to get turn down service despite telling them we went to bed early (if you consider 10 pm early)
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|Children's Programs
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|Cabin / Stateroom
|Do NOT stay here with 4 people! It is way to small.
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|New Orleans, Louisiana
|Add a tip. What was your favorite thing to do here?
|Cozumel, Mexico
|Chankanaab was great
|Belize City, Belize
|Starfish island was not great, but ok. Beach is rough and water is murky.
|Roatan (Isla Roatan), Bay Islands, Honduras
|Snorkeling was incredible
|Costa Maya (Mahahual), Mexico
|Add a tip. What was your favorite thing to do here?
Pizza very hard to get and not that good, but ice cream and chili dogs were really good.
5 Comments
How strange that NCL, which started freestyle dining, can't seem to get those lines eliminated. I do not know how you manage four passengers in a inside room - especially since two are teens. Nice to read, though that you enjoyed the food and the shows. Those are the two of the most important reasons that I take cruises. The third is the itinerary.
We did an interior cabin on Carnival twice with my family and this cabin just seemed very small when comparing to the Carnival ships we have been on. We will definitely cruise again, but I am sure I we will get a much larger cabin and focus more on the things on the ship for our kids. The big thing was that this particular ship did not have a mini-golf course or water slide or any other feature that our kids were excited about, and this was spring break, so the pool and hot tubs were overly full of college kids, so my teens were bored.
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Reviewed By Kyle H (Golden, CO) Reviewed for a 2009 Pontiac Vibe —7/12/2012 8:13 AM
I love my new sun shade, it's easy to use and keeps my car much cooler.
Reviewed By PATRICIA C (Murrieta, CA) Reviewed for a 2009 Pontiac Vibe —4/17/2007 10:08 PM
Love this sunshade - complete coverage and good fit. Much better than generic shades that don't fit properly, especially on a large windshield. Protects interior and keeps vehicle cooler in hot weather.

subset: commoncrawl


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The 35-year-old soldier, from the 2nd Royal Tank Regiment, died from injuries caused by an improvised explosive device in Basra, southern Iraq, on Tuesday night.
Cpl Edwards, known as Eddy to his friends, was on a routine patrol in the Mustashfa district of Basra City when his Warrior armoured vehicle was struck shortly before midnight local time. The latest death brings the total number of British service personnel who have died in Iraq since the 2003 invasion to 164.
The MOD said Cpl Edwards was checking the safety of the soldiers travelling in his vehicle at the time of the attack. He had been serving in Iraq since May and was often involved in operations in Basra City.
A member of Badger Squadron, 2nd Royal Tank Regiment, attached to the Irish Guards Battle Group, Cpl Edwards was working under the command of 4th Battalion The Rifles.
Cpl Edwards, from Sutton Coldfield, joined the Army in 1993 and had deployed on exercises all over the world. As well as serving in Iraq, he served on operations in Northern Ireland and the Balkans. He was due to be promoted to the rank of sergeant later this year at the end of his tour in Iraq.
Cpl Edwards leaves a wife, Gabriele, and an 18-month-old son, Ryan.
Lieutenant Colonel David Catmur, commanding officer of 2nd Royal Tank Regiment, praised Cpl Edwards, with whom he had worked for a number of years.
"He was a cheerful, enthusiastic man who was very proud to be a part of our regimental family," he said. "He was a 'Tankie' to the core: proud, loyal and professional with an excellent esprit de corps.
"His years of experience and kind-hearted nature will be a great loss to us all. Eddy epitomised all that is good about our soldiers, an example to all. Our thoughts now go to his wife, Gabriele, his son, Ryan, and his parents and family."
- More about:
- Basra
- Iraq
- Military
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The Women's Namella by Fila was designed with features that allow optimal breathability and impact absorption. These sneakers feature elasticized and breathable fabric uppers, memory foam cushioned comfort insole, and lightweight EVA traction outsole. This sneaker takes comfort to the next level so you can get the most out of your day.
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Hulne Priory
History, tourist information, and nearby accommodation
One of the unusual features at Hulne is that the monastic buildings are contained within a retaining wall, a reminder that during much of the medieval period Northumberland was a battleground between Scots and english. In the late 15th century the monastery defenses were improved by adding a sturdy pele tower, a form of fortified lookout, allowing the monks to retreat inside the tower if truble threatened.
Much of the priory is in ruins, though the infirmary walls have been restored and now stand to their full height, though there is no roof. There is a well-preserved triple sedilia in the priory church, with very finely carved arcading.
After the priory was dissolved the property fell to the Percy family, Dukes of Northumberland, and the priory buildings now stand amidst Hulne Park, used by the Dukes as a hunting park. A very short walk away is Alnwick Abbey Gatehouse, the only surviving part of a 12th century Premonstratensian abbey.
Getting there
Access is only on foot, from Alnwick. Park on the left side of Rotten Row, off the B6346 road to Wooler. Access to the Park is limited, at this writing 11-sunset, but it is worth checking first.
About Hulne Priory
Address: Hulne Park, Alnwick, Northumberland, England
Attraction Type: Abbey
Location: From Alnwick take Ratten Row, off the B6346 Wooler road. Limited parking along the verge for foot access to Hulne Park. Public access to the park is permitted, but there are no visitor facilities.
Website: Hulne Priory
Location map
Photo Credit: Les Hull, licensed for reuse under the Creative Commons Licence
We've 'tagged' this attraction information to help you find related historic attractions and learn more about major time periods mentioned.
Historic Time Periods:
Find other attractions tagged with:
12th century (Time Period) - Henry VIII (Person) - Victorian (Time Period) -
Heritage Rated from 1- 5 (low-exceptional) on historic interest
Alnwick Abbey Gatehouse - 1.3 miles (Abbey)
Bailiffgate Museum - 1.8 miles (Museum)
Alnwick Castle - 2 miles (Castle)
Alnwick Garden - 3.9 miles (Garden)
Edlingham Castle - 4.9 miles (Castle)
Edlingham, St John the Baptist Church - 5.1 miles (Historic Church)
Preston Tower - 6.2 miles (Historic Building)
Dunstanburgh Castle - 6.7 miles (Castle)
Nearest Accommodation to Hulne Priory:
Nearest Self Catering Cottages
Alnwick, Northumberland
Sleeps: 6
Stay from: £277.00 - 747.00
Alnwick, Northumberland
Sleeps: 7
Stay from: £275.00 - 835.00
Eglingham, Northumberland
Sleeps: 5
Stay from: £425.00 - 1230.00
More self catering near Hulne Priory >>
Nearest Hotels
Alnwick, Northumberland
Stay from: £120.00
Alnmouth, Northumberland
Stay from: £50.00
Newton-on-the-Moor, Northumberland
Stay from: £50.00
More Hotels near Hulne Priory >>
Nearest Bed and Breakfasts
Alnwick, Northumberland
From: £50.00
Alnwick, Northumberland
From: £59.00
Alnwick, Northumberland
From: £70.35
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Fabricio Coloccini is an Argentine footballer who plays for English club Newcastle United and the Argentina national team. Coloccini has had an eventful career playing for several different clubs including Boca Juniors, AC Milan, Deportivo la Coruna and Newcastle United.
Coloccini was born on the 22nd of January 1982 and he began his club career at Boca Juniors in 1998. He made ust two appearances scoring one goal before moving to Europe with AC Milan. Coloccini found it difficult to break through to the first team and he went through a series of loan spells and San Lorenzo, Alaves, Atletico Madrid and Villarreal from 2000 to 2004.
In 2004 he signed a permanent deal with Deportivo la Coruna on a six year deal and during his three and a half year stint he helped the club to win the 2008 UEFA Intertoto Cup. In 2008 he signed for English club Newcastle United for £10.3 million. He made his club debut in a 1-1 away draw against Manchester United. Since his debut he has been one of the most consistent players in the team.
Newcastle were relegated in the 2008-09 season and helped the club to gain promotion to the Premier League in the 2009-10 season where the club won the Championship. In July 2011 Coloccini was named club captain for the 2011-12 season and he formed a unique partnership with Steven Taylor for the following few seasons. Coloccini made his debut for Argentina in 2003 and played in the 2006 FIFA World Cup and he helped the club to gain promotion to the 2014 FIFA World Cup in Brazil.


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Reviewed by 1 customer
Best Uses
Comments about Spirit ThunderCats Lion-O Adult Mens Costume:
It was a great costume. I just wish it would have been tighter. The belt and the sword are a must have for this outfit to work. Withoug the two, the younger generations just wont have a clue as to who you are.


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Prezzo - Leicester
22 Silver Street, Leicester, LE1 5ET +44 1162389764
- Genomsnittlig
- Huvudrätt: £9
- Matkostnad: £17
- Öppettider
- Sunday to Thursday: 11:30 - 22:30 Friday to Saturday: 11:30 - 23:00.
Always , inviting , food wonderful , staff very nice .
Very clean
Carol Love
06 november 2015
från Leicester
Buy a dining gift at your favourite Prezzo restaurant…
3 courses & a glass of wine, £30 for 2 people
3 courses & a bottle of prosecco, £55 for 2 people
Prezzo - Leicester serves delicious Italian cuisine in a contemporary and stylish setting. Leicester Rail Station is only an 11-minute walk from Prezzo.
The Prezzo menu presents a range of exciting dishes, crafted using the season's best ingredients. Sample meat & fish dishes like chicken Milanese or sea bass, along with classic pasta recipes, hand-stretched, stone-baked pizzas, and risotto.
All the favourite antipasti dishes are on offer, as well as speciality folded calzones, and piled-high 'la famiglia' pasta dishes, designed for sharing. Start your visit with a fruity, fizzy pre-dinner aperitivo from the Prezzo cocktail list, and finish with an indulgent Italian-style dessert.
Leicester Prezzo has a modern, relaxing ambience, and Leicester Cathedral and Curve Theatre are within easy walking distance.
Prezzo - Leicester
22 Silver Street, Leicester, LE1 5ET
Se alla Prezzo Restaurants
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Se alla restauranger i Leicester
Kundbetyg för Prezzo - Leicester
- Mat
- Värde
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- Plats
- Service
- Drycker
(**** 14 november 2015 av Simon från Leicester
Food was good but service was only ok. Despite the restaurant not being full, we were put on a table that was squashed around the back of the stairs which would have been ok except that we seemed to be forgotten after a while. First two courses were quick and good but there was a long wait following that and after dessert we were completely forgotten.
The restaurant itself is very well decorated but the toilets are terrible. Very tired and in much need of attention. Overall, it was only OK and I don't think we'll return.
((((( 6 november 2015 av Carol Love från Leicester
Always , inviting , food wonderful , staff very nice .
Very clean
((((( 27 oktober 2015 av Alison morris från Leicester
Great food, buzzing and vibrant, good choice on menu - will definitely be back
((((* 4 oktober 2015 av Sara Towers från Leicester
delicious food, waiter a little annoying and ott
((((( 16 augusti 2015 av Margaret Collick från Derbyshire
Excellent food. We are regulars. Lovely atmosphere. The manager is very funny. We will be back again.llove
((((* 18 juni 2015 av Eileen Wilford från Leicester
Meal was hot and served well we had a really relaxing lunch, which is has always been the case on my previous visits
((((* 17 juni 2015 av Sharon Simpson från Leicester
Very good service and good value
((((( 29 april 2014 av Maxine Hough från Leicester
Great meal, courteous service, a lovely ambience.
((((( 28 april 2014 av Karl Madelin från Leicester
Great service and food
((((* 8 april 2014 av Priya Pancholi från Leicester
Lovely at prezzo as always
Tyvärr så kan du inte boka Prezzo - Leicester online via vår hemsida
Befintliga erbjudanden
Erbjudanden direkt till din inkorg!
Hör om våra topperbjudanden först, och boka de bästa borden.
Du kommer snart att få tillgång till en massa bra erbjudanden
Restauranger i närheten
106m 63 recensionerASK Italian - Leicester
ASK Italian - Leicester
206m 0 recensionerCoast to Coast - Leicester
Coast to Coast - Leicester
284m 11 recensionerByron - Leicester
Byron - Leicester
288m 4 recensionerChimichanga - Leicester
Chimichanga - Leicester


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Each month, Rated People connects over 50,000 homeowners with quality tradesmen - simply enter your job details in the form above to get started.
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After the work is complete, return to the site and rate your tradesman based on their quality of work, reliability and value for money.
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Steam clean a small terraced house
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carpet clean (there is carpet every room except kitchen and batchroom), dusting, light lint...
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via DNA:
Bubba the Love Sponge has implied Hulk Hogan leaked his own sex tape.
The DJ lashed out at his former best friend – who can be seen cavorting with Bubba's estranged wife Heather Clem in the video – and thinks the "self-centred" wrestler could have released the footage for publicity.
Speaking on his radio show, Bubba – who is believed to feature at the end of the tape bragging about how he could sell the footage – said: "I'm gonna be honest with you, I have not seen the tape … including me supposedly bragging and being a horse's ass.
"At the end of the day, I can't even comment on the validity … [Hulk] could have had something to do with the leak of it."
Hulk is suing Bubba and Heather for $100 million, claiming they filmed him without his consent.
However, Bubba issued a reminder to Hulk live on air, saying: "Nobody put a gun to your head" and insisted it was consensual.
He also lashed out at his former friend, defending the wrestler' ex-wife Linda and calling Hulk's daughter Brooke talentless.
The 'TNA IMPACT Wrestling' star's sex tape was previously thought to have been leaked by a disgruntled ex-employee of Bubba.
Hulk recently called in the FBI because he is determined to get to the bottom of the violation of his privacy.
He insisted, "I have braced my family, there could be another one. I have no idea. I've never made a tape. I swear to god I wasn't in on this."


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No longer designated a youth ensemble, the flagship orchestra of Venezuela's revolutionary educational initiative El Sistema comes to Carnegie Hall as part of the Voices from Latin America festival. Conducted by the exuberant Gustavo Dudamel (the world's most notable Sistema graduate), the orchestra plays works by Carlos Chávez, Julián Orbón and Silvestre Revueltas on Mon 10, and by Heitor Villa-Lobos and Antonio Estévez on Tue 11.
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Dear customer, kindly fill in your name, contact number and email, so we can inform you when the new stock of this product arrive.
Email your friend about this product
Superbly molded in coloured plastic, with the same fantastic construction quality demonstrated in previous PG kits.
A whopping number of parts are included in the typically-large (and heavy!) box.
Besides the Strike Rouge, there are parts included for the Skygrasper and Aile Striker!
Metal-plated parts (in pink!) are included for the Strike Rouge's custom weapons.
Screws, springs, fabulous Gundam Stickers and dry-transfer decals are included; two construction manuals (one for the Strike Rouge, one for the Skygrasper and Aile Striker) are included to walk you through the building of all three models.
An "instruction manual" is also included for those who like to paint their Gundam models (of course it doesn't need to be painted), and also for indicating precise locations of the markings.
We currently charge shipping fees on 'Per Order' basis. Depending on your location and shipping mode the price will vary, please refer to chart below for details.
All our orders are packed and shipped within 24 hours upon payment confirmation. We use POSLAJU / Air Parcel / TA-Q-BIN / ABX for most of our orders . They are able to ship overnight to most cities and towns in the Peninsular Malaysia, however you should allow 1-3 business days in case any unexpected delays. East Malaysia orders may often take 4 to 5 extra days, so please allow 5 to 7 business days for delivery.
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Speaking to journalists after receiving an award at an event organized by the Petroleum Products Pricing Regulatory Agency (PPPRA) in Abuja yesterday, Kachikwu said;
"First, I am not aware that the NNPC has increased price. I need to look into that. It is a bit of surprise for me, because there are processes in doing this, if they have done that, it means they are doing it wrongly. Let me find out what the facts are".


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Spain`s Pedrosa takes MotoGP pole at Indianapolis
Indianapolis: Dani Pedrosa had the fastest motorcycle in the rain and was even quicker on a dry track.
It`s still not good enough for the 23-year-old Spaniard.
So Pedrosa has asked his Repsol Honda team to make more adjustments after outdueling his top two rivals Saturday to take the Indianapolis MotoGP pole with a fast lap of 1 minute, 39.730 seconds. Spain`s Jorge Lorenzo was second at 1:40.236, with six-time world champ Valentino Rossi third in 1:40.609.
"I`m happy," Pedrosa said. "I think we have to look for something else. I think we always should have to work and try to improve for tomorrow."
Most expected Rossi and Lorenzo, the Fiat Yamaha teammates, to continue their season-long dominance.
Pedrosa hasn`t given them a chance.
He topped the speed charts Friday afternoon, in the rain, and again Saturday morning, on a green track. Then in the 1-hour afternoon qualifying session, he rode on the edge, occasionally tapping the surface with his knee pads yet maintaining enough balance to zip through the corners and the straightaways on the 2.621-mile road course more than a second faster than Rossi did in last year`s inaugural race.
The result: Pedrosa`s speed of 94.56 mph was a 10 mph improvement from Friday.
Italy`s Rossi, the defending race champ and current points leader, and Lorenzo couldn`t keep up.
That ended a four-race streak in which Rossi or Lorenzo had won the pole, and it`s only the fourth time this year one of the teammates has not had the pole. Rossi is making his fourth start of the season outside the top two spots and acknowledges his motorcycle may be quick enough to catch Pedrosa.
"Honestly, we don`t have enough pace at the moment, and I can`t ride how I want to," Rossi said. "We don`t have quite enough grip to enable us to push at the maximum. We did a lot of work this afternoon, but we`re still not at our best."
That`s certainly not the case for Pedrosa, who claimed his second pole of the season and eighth in his MotoGP career.
Pedrosa knocked 11 seconds off his best time from Friday in the morning practice and shaved another 1.5 seconds off that pace in qualifying.
Now he can go for an American sweep after winning in California in July and perhaps even become the first motorcycle rider to hit 200 mph down Indy`s famed straightaway. Track officials said speeds topped 197 on Saturday and with the forecast calling for cooler temperatures and drafting expected under race conditions, it is possible.
"If the wind changes, maybe we can go faster than that in the trap speed," Pedrosa said.
Thirty minutes into qualifying session, Lorenzo was clocked in a record 1:40.643. Five minutes later, Pedrosa jumped to the top of the speed charts by going 1:40.440.
In the final 10 minutes, three riders — Lorenzo, Rossi, and San Marino`s Alex De Angelis — all broke 1:41.
Pedrosa outdid that, too, breaking 1:40 on two of his last three laps — a first — to win the pole.
De Angelis starts fourth after going 1:40.620.
Colin Edwards was the top American qualifier at 1:40.961. He`ll start fifth.
Nicky Hayden, the 2006 world champ who lives in Kentucky, was sixth in 1:41.067.
"I still feel I could have gone faster at the end, and I`m not completely happy," Hayden said. "I know what I`m in for tomorrow, and I`m not expecting to find a full second in the morning (practice)."
Three drivers, including Edwards, slid off their motorcycles during qualifying, but none appeared seriously injured. All three walked away from the accidents.
But Pedrosa isn`t leaving anything to chance.
"I`ve been comfortable on the machine from the start and things have just clicked here," he said. "We can`t relax at all, though, because I`m sure the other riders will improve and we know how strong our rivals have been this season. So we must make a few small adjustments to make sure we have the best possible package."
Bureau Report


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EURUSD: Maintains Recovery Tone, Further Upside Envisaged
Although the pair closed marginally higher the past week, EUR continues to hold on to its recovery tone set from the 1.3144 level. This leaves scope for further strength towards the 1.3937 level, its Sept 15'2011 low. Further out, resistance stands at the 1.4283 level, its Sept 06'2011 high. And possibly higher towards the 1.4498 level. Its weekly RSI is bullish and pointing higher suggesting further strength. Alternatively, the risk to our upside view will be a return to its Sept 28'2011 high at 1.3690 and its Oct 18'2011 low at 1.3650. We may see a reversal of roles as support at this level turning EUR higher but if taken out, risk will build up on the 1.3377 level, its Oct 10'2011 low and subsequently the 1.3144 level. All in all, EUR continues to build on its recovery strength started from the 1.3144 level.


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Tight junctions and the modulation of barrier function in disease
- First Online:
- Accepted:
DOI: 10.1007/s00418-008-0424-9
- Cite this article as:
- Förster, C. Histochem Cell Biol (2008) 130: 55. doi:10.1007/s00418-008-0424-9
- Citations
- 3.4k Downloads
Tight junctions create a paracellular barrier in epithelial and endothelial cells protecting them from the external environment. Two different classes of integral membrane proteins constitute the tight junction strands in epithelial cells and endothelial cells, occludin and members of the claudin protein family. In addition, cytoplasmic scaffolding molecules associated with these junctions regulate diverse physiological processes like proliferation, cell polarity and regulated diffusion. In many diseases, disruption of this regulated barrier occurs. This review will briefly describe the molecular composition of the tight junctions and then present evidence of the link between tight junction dysfunction and disease.
KeywordsTight junction Occludin Claudin Cancer Inflammation Toxin Hereditary Vascular
Tight junctions (TJ) (zonulae occludentes) form a continuous, circumferential, beltlike structure at the boundary between the apical and the basolateral membrane domains in epithelial and endothelial cells. By constituting a regulated diffusion barrier for the paracellular pathway, TJs establish separate compartments in multicellular organisms and are also crucial for the exchange of substances between the internal and external cellular environment by the expression of tissue-specific transport proteins and channels. The transmembrane proteins constituting the TJs are attached to the cytoskeleton, thereby linking cell–cell and cell–substratum adhesion sites. In addition, cytoplasmic plaque proteins constitute scaffolds for TJ assembly or are involved in the regulation of processes like transcription, proliferation and differentiation into a tissue-specific regulated diffusion barrier in physiology and development. Many excellent reviews have summarised data regarding the molecular composition and function of TJs (Ebnet 2008; Gonzalez-Mariscal et al. 2003; Matter and Balda 2003a; Mitic and Anderson 1998; Schneeberger and Lynch 2004; Tsukita and Furuse 1999; Zahraoui et al. 2000), so that the scope of the current review is focused on disturbances of TJ function in human diseases.
Structure of the tight junction
The claudins
Searching for the proteins forming the structural backbone of the TJ strands, the Tsukita group re-examined the junctional fraction prepared from chicken liver and discovered the first members of the claudin family: two 22-kDa proteins, claudin-1 and claudin-2 (Furuse et al. 1998a).
The claudins also have four transmembrane domains, but do not show any sequence similarity to occludin (Fig. 3b). So far, 24 members of the claudin family have been identified in different species. Claudins appear to be expressed in a tissue-specific manner; some claudins are expressed only in specific cell types, e.g. claudin-5 is expressed primarily by vascular endothelial cells or claudin-11 appears to be expressed solely in oligodendrocytes and Sertoli cells (Morita et al. 1999). Moreover, most cell types express more than two claudin isoforms conferring different size and charge selectivity qualities by the amino acid sequence of their extracellular loops, and the combination of those isoforms results in cell- or tissue-type specific barrier function by "heteropolymerisation" into the resulting TJ strands (see below). As demonstrated by immunogold electron microscopy, there is accumulating evidence now that the claudins do constitute the TJ strands as observed by freeze–fracture electron microscopy (Tsukita et al. 2001). Exogenous expression of claudins in fibroblasts devoid of endogenous TJs resulted in the formation of TJ strands.
Summarizing existing data, there is clear evidence that the claudins constitute the backbone of TJ strands, while occludin, which itself cannot reconstitute such TJ strands, seems to play a permeability regulating role by incorporating itself into the claudin-based strands, a process, the mechanism of which however is not yet understood. The function(s) of occludin thus still remain to be elucidated.
Junction-associated adhesion molecules (JAMs)
A third class of integral membrane proteins comprises members of the immunoglobulin superfamily (Fig. 3c), and can be subdivided into a group consisting of JAM proteins and a group consisting of CAR, CLMP, ESAM and JAM-4 (Ebnet 2008; Ebnet et al. 2004). To date, at least three JAM isoforms have been described, which are expressed differentially in epithelial and endothelial cells, but also in cells devoid of TJ strands like leukocytes (D'Atri and Citi 2002). Except for JAM-1, little is known about the role of these proteins in TJ assembly and function. JAM-1, formerly F11 receptor (Sobocka et al. 2000) (Fig. 3c), may be involved in immune cell transmigration or cell adhesion (Bazzoni et al. 2000) but has not been localized to the junction strands. A putative PDZ binding motif in the cytoplasmic domain of JAM-1 has been identified, which has been able to interact with PDZ domains of ZO-1, AF-6, PAR-3 and MUPP1 (D'Atri and Citi 2002; Itoh et al. 2001). Moreover, JAM-1 has been shown to interact with cingulin. Thus, even though JAM-1 is not part of the TJ strands, it may well be involved in the propagation of signal cascades resulting from homophilic and heterophilic adhesion. A role for JAM-1 in the complex process of adhesion and transmigration of monocytes through endothelial cells has been demonstrated (Martin-Padura et al. 1998). As another function for the JAM proteins, a role in the assembly of TJs and regulation of paracellular permeability could be deciphered by ectopic transfection experiments (Cohen et al. 2001). Interaction of JAM-1 with occludin, so far not determined whether direct or indirect in nature, has been shown (Liu et al. 2000).
TJ plaque proteins and the coordination of signaling at the TJ
In addition to their prime function as a regulated permeability barrier in the paracellular pathway and as fence in the plane of the membrane, TJs play a pivotal role in organizing such diverse processes as morphogenesis, cell polarity, cell proliferation, and differentiation, which require the coordination of signals impinging on and emanating from the plasma membrane. Mounting evidence suggests that the cytosolic TJ plaque is one of the sites in which such signaling is coordinated (Ebnet 2008; Gonzalez-Mariscal et al. 2003; Schneeberger and Lynch 2004). Since identification in 1986 of zonula occludens (ZO)-1 as the first TJ-associated protein (Stevenson et al. 1986), almost 30 additional proteins have been described associated with the cytoplasmic aspect of TJs (Schneeberger and Lynch 2004). They can be grouped into two major categories: The first are the peripherally associated scaffolding proteins like ZO-1 (ZO-2, ZO-3, AF6, and cingulin) that appear to organize the transmembrane proteins and couple them to other cytoplasmic proteins and to actin microfilaments. The second are numerous "signaling" proteins (ZONAB, RhoA, RalA, and Raf-1) proposed to be involved in junction assembly, barrier regulation, gene transcription, and perhaps other, presently undefined pathways.
TJs as permeability barrier
One of the key functions of tight epithelia is the creation of a diffusion barrier by tight control of the transcellular and paracellular pathways. This is acomplished by asymmetrical distribution of transporters and channels for the transcellular route, and on the other side by regulation of diffusion through the paracellular space via integral TJ proteins (Turner 2006). Hereby, certain claudins participate in the formation of ion-selective pores: available data suggest that TJs on the one hand are permeable to small noncharged solutes up to a size of 0.43–0.45 nm in radius as shown by the use of the membrane-impermeant polyethylene glycol as a tracer, but investigations also showed that pores within the TJs are capable of discriminating between ion charge and size, which both are additionally dependent on concentration and pH (Schneeberger and Lynch 2004; van Bree et al. 1988).
The two extracellular loops of occludin consist solely of uncharched amino acid side chains (Fig. 3a) represented mainly by an unusually high content of tyrosine and glycine (Furuse et al. 1993) so that a direct role for occludin in conferring charge selectivity is doubtful. However, occludin could increase electrical resistance by interaction with the extracellular domains of the differently charged residues of the different claudins (see below), since ablation of occludin is not without consequence despite the fact that the protein does not appear to be required for TJ strand formation. Occludin-null mice express a complex phenotype, which is, however, not manifested by structural or functional TJ abnormalities: in occludin −/− mice, TJs were not affected morphologically, and electrophysiological barrier function of intestinal epithelium was normal. However, histopathological abnormalities were detected in several tissues, i.e., chronic inflammation and hyperplasia of the gastric epithelium, calcification in the brain, testicular atrophy, the loss of cytoplasmic granules in striated duct cells of the salivary gland as well as the thinning of the compact bone, pointing to complex functions of the occludin protein (Saitou et al. 2000).
Role of claudin proteins in formation of a TJ permeability barrier
The lack of charged amino acid residues within the extracellular loops of occludin segregates occludin from the candidates contributing directly to the formation of ion-selective pores. The different claudins whose extracellular loops span a wide range of pKion the contrary appear to be excellent candidates to mediate ion selectivity in TJ permeability (Fig. 3b). Major insight into the distinct charge selectivity of the claudins came from the work of Furuse et al. (2001), who could relate the different levels of resistance between MDCK I cells (TER > 10,000 Ωcm2) and MDCK II cells (TER <200 Ωcm2) to a differential claudin isoform expression, while MDCK I cells express claudin-1 and -4 but not claudin-2, MDCK II cells additionally express claudin-2, which seems to mediate higher ion permeability by pore formation/inducing strand breaks. Notably, transfection of the tight MDCK I with claudin-2 cDNA resulted in 20-fold decline in TER but not in the number of TJ strands.
A wide range of further transfection and mutagenesis experiments introducing or ablating different claudins has been performed to date, and been partially helpful to elucidate the conductance conferring properties of the resulting pores (Matter and Balda 2003b). These informative observations must however be viewed with some caution because transfection and site-directed mutagenesis may lead to more than a change in net charge as for example an altered molecular conformation, or altogether the introduction of an unusual claudin protein could lead to disturbance of strand organisation within native strands. By the construction of chimera, Colegio et al. (2003) were moreover able to attribute ion selectivity of the claudins to distinct stretches within the primary amino acid sequence of the extracellular loops leading to alterations in TER and charge selectivity in transfection experiments. Taken together, available data strongly suggest a key role for the extracellular loops of the diverse claudins in the formation of ion selective pores within the TJ strands.
Role of occludin in formation of a TJ permeability barrier
The role of occludin in TJs and a putative contribution to building up the TJ's ion selectivity (the ion pores) is unclear; it has been demonstrated that overexpression of occludin in MDCK cells reduces ion permeability but in the same time was shown to increase the transepithelial flux of the uncharged compound mannitol (Balda et al. 1996). Further observations then implicated that occludin may be a target for receptor-initiated signalling involving the Rho family of GTPases, and that by an additional not-yet-understood mechanism, occludin phosphorylation may regulate TJ permeability independently of cytoskeletal activity (Hirase et al. 2001).
Some indirect evidence for a role of occludin in strengthening barrier properties arose from heterologous expression studies: when expressed in L-fibroblasts that lack endogenous cadherin-based adherens junctions and ZO-1, little adhesiveness was measured (Kubota et al. 1999). However, when overexpressed in fibroblasts, which do have well developed ZO-1-containing adherens-like junctions (NRK-fibroblasts), increased Ca2+-independent adhesion was observed, indicating that when interacting with ZO-1, occludin mediates adhesive functions (Van Itallie and Anderson 1997). Addition of neutralizing peptides of the peptide sequence of the second extracellular loop elicited competitive interference with cell–cell-adhesion in a dose dependent manner, implicating that this loop directly participates in adhesive interaction (Wong and Gumbiner 1997).
Relating epithelial malignancy to the loss of cell–cell-adhesion, it was demonstrated that overexpression of oncogenic Raf 1 in rat salivary gland epithelial cells (Pa4–Raf1) disrupts TJs and induces an oncogenic phenotype by downregulating the expression of occludin (Wang et al. 2005). Raf1-mediated transformation of Pa4–Raf1 cells was subsequently shown to induce transcriptional downregulation of occludin and forced re-expression of occludin rescued the polarized phenotype of epithelial cells (Wang et al. 2007). Interestingly, forced expression of an occludin mutant lacking the second extracellular loop did not rescue the epithelial phenotype in vitro nor did it prevent tumor growth in vivo. These results demonstrate that the TJ protein occludin has a potent inhibitory effect on the Raf1-mediated tumorigenesis, and the second loop of occludin appears to be required for suppression of Raf1-induced tumor growth, potentially by mediating cell–cell-adhesion as presented above contributing to contact inhibition of cell division.
TJs and human disease
Modulation of barrier function by cytokines
Modulation of TJ barrier function in human disease
Tight junction protein affected
Breast cancer: invasive ductal cancer
Claudin-1, ZO-1
Itoh and Bissell (2003), detailed here
Prostate cancer: prostatic adenocarcinomas
Claudins-1, -3, -4 and claudin-7
Sheehan et al. (2007)
Thyroid neoplasma, follicular adenoma
Occludin and claudin-1, -4 and -7
Tzelepi et al. (2007)
Gastroesophageal reflux disease: Barrett's esophagus (dysplasia)
Jovov et al. (2007)
Lung cancer: Basaloid squamous carcinoma
Ultrastructural junctional dedifferentiation
Gilcrease and Guzman-Paz (1998)
Inflammatory bowel disease: Morbus Crohn
Claudins-2,-3, -5, and -8, ZO-1
Zeissig et al. (2007), detailed here
Collagenous collitis
Occludin, claudin-2 and -4
Burgel et al. (2002)
Multiple sclerosis
Occludin, claudin-5
Förster et al. (2007), detailed here
Hereditary diseases
Hereditäry deafness
Ben-Yosef et al. (2003), detailed here
Familial hypomagnesemia
Simon et al. (1999), detailed here
cystic fibrosis
Occludin, claudin-1, -4, JAM, ZO-1
Coyne et al. (2002)
Vision loss
Diabetic eye disease: diabetic retinopathy
Occludin, ZO-1
Felinski and Antonetti (2005), detailed here
Viral infection
Reoviral infection (hydrocephalus, encephalitis)
Forrest et al. (2003)
Bacterial toxins
Clostridium perfringens enterotoxin
Claudin-3 and -4
McClane (2001), detailed here
Inflammatory bowel disease: Crohn's disease
Crohn's disease ranks among the chronic inflammatory bowel diseases (IBD) with diarrhoea as a leading symptom. This is chiefly attributed to epithelial barrier dysfunction, leading to an increased loss of solutes in the form of "leak-flux diarrhoea". Notably, the integrity of the intestinal barrier is not only impaired in patients with Crohn's disease but also many first-degree relatives of Crohn's disease patients present with increased intestinal permeability in the absence of clinical symptoms (Hollander 1993; Peeters et al. 1997; Yacyshyn and Meddings 1995). From this observation it was concluded that barrier dysfunction in Crohn's disease precedes the inflammatory process in contrast to many other inflammatory diseases, probably arising from genetic defects or driven by environmental factors like intestinal luminal antigens or bacterial toxins.
The epithelial barrier in the intestine is established by a single layer of epithelial cells. However, barrier dysfunction alone does not to seem be sufficient to cause Crohn's disease, since unaffected relatives of patients with Crohn's disease comparably present increased permeability, but do not show the abnormal mucosal inflammatory response reported of patients with Crohn's disease (Weber and Turner 2007). In healthy individuals, the intestinal mucosal barrier is organised through tight junction strands connecting the adjacent epithelial cells (Weber and Turner 2007). These TJs seal the paracellular space and an intact intestinal mucosa. TJ permeability is rate limiting for epithelial permeability and defects thereof are an established source for intestinal barrier defects as observed in Crohn's disease. The basic understanding of TJ function we developed is that they form an ion-selective and size-selective paracellular gate whose permeability varies throughout the gastrointestinal tract (see above). According to this, it is not surprising that the pattern of claudin isoforms expressed varies alongside the gastrointestinal tract (Nusrat et al. 2000). Recent studies now show that the expression pattern of characteristic claudins is altered in Crohn's disease (Weber and Turner 2007) with a concomitant effect on the integrity of TJ strands: Zeissig et al. (2007) showed by freeze–fracture electron microscopy that the overall number of TJ strands sealing the epithelial layer is reduced while concomitantly an increased number of strand breaks was noted in tissue from patiens with Crohn's disease (Zeissig et al. 2007). The expression of 12 claudin isoforms in the intestinal epithelium was investigated in healthy individuals and patients with Crohn's disease, leading to the insight that the expression of the claudins-3, -5, and -8 was decreased in active disease. On the contrary, expression of barrier-reducing claudin-2 was increased in histological samples from patients, which was particularly observed in the crypt epithelium. This altered pattern of claudin isoform expression was thus concluded to be partly responsible for the observed mophological and functional disruption of TJs. However, claudin isoform expression patterns were unaltered in patients with inactive Crohn's disease, rendering this observed change in expression patterns rather a consequence than a cause of active disease.
It is has been acknowledged for a long time that chronic inflammation in Crohn's disease is associated with the overproduction of proinflammatory cytokines. Searching for the nature of these pro-inflammatory mediators effecting the TJ disruption and dysfunction, investigators (Conyers et al. 1990; Heller et al. 2005; Prasad et al. 2005) had identified IL-13, IFNγ and TNFα as factors in patient's serum lowering resistance and opening TJs. However, differential effects of the different cytokines crititical to IBD pathogenis have been reported from studies in cell culture models, indicating that the mechanisms by which IFNγ and TNFα inrease paracelluar permeability differ from those used by IL-13 (Prasad et al. 2005). IFNγ and TNFα exposure of model cell lines led to a reduction in claudin-2 expression, while IL-13 exposure induced increased claudin-2 expression. Those observed discrepancies, which might be based on the source of epithelial cell lines, the experimental design or the doses of cytokines applied are the focus of current investigation. Other lines of research are concentrated on gaining more information on the source of pro-inflammatory cytokines in IBD and the mode how those cytokines may work together to modify claudin expression pattern, either in vitro or in vivo (Weber and Turner 2007).
At the cell biological level, recent investigation demonstrated that IFNγ exposure promoted the endocytosis of occludin, JAM-1 and claudin-1 into early and recycling endosomes (Bruewer et al. 2005), an effect which appears to be mediated by RhoA/Rho kinase, which further induced a myosin II-dependent formation of vacuoles in the apical cell moiety (Utech et al. 2005).
At the gene expression level, other research groups could demonstrate that TNFα induced an increase in Caco-2 TJ permeability mediated by activation of the pro-inflammatory transcription factor NFκB. NFκB action leads to a downregulation of specifically ZO-1 gene expression and a redistribution of ZO-1 protein away from the intercellular junctions (Ma et al. 2004). A comparable effect could be described in the nonintestinal model epithelial cell line, MDCK, and in the blood brain barrier model cell line cEND, where also disruption of TJs following exposure of epithelial and endothelial cells to TNFα could be demonstrated (Förster et al. 2007; Poritz et al. 2004).
Blood-brain barrier compromise in neuroinflammation–multiple sclerosis
Homeostasis of the central nervous system (CNS) microenvironment is essential for its normal function. It is maintained by the blood-brain barrier (BBB) which regulates the transport of molecules from blood into brain and backwards. The function of this highly specialised barrier is to (1) protect the brain from blood-borne substances and (2) to provide nutrient supply to the brain parenchyme by specialised transport systems (Rubin and Staddon 1999).
The main structures responsible for this barrier property are the TJ. TJ are strongly developed in endothelial cells of the BBB but only moderately formed between endothelial cells of the peripheral vasculature: leaky blood vessels in the body allow many molecules to cross through to tissue, but the tight construction of the vessels in the CNS guards against brain entry, leading to high electrical resistance in the range of 1,500—2,000 Ωcm2, depending on the species investigated. In BBB-forming brain capillary endothelial cells (BCECs) the TJ proteins occludin and claudin-5, claudin-3, claudin-1 and claudin-12 were demonstrated by immunofluorescence and western blot analysis (Förster et al. 2007; Wolburg and Lippoldt 2002).
Under physiological conditions, brain microvascular endothelial cells form a barrier limiting the extravasation of cells of the immune system like monocytes, lymphocytes and other leukocytes. It has been shown in this context that leukocytes can transmigrate to the BBB without net changes in vascular permeability and cell–cell contacts (Cramer 1992). However, increased leukocyte migration as in multiple sclerosis (MS) has been reported to lead to a re-organisation of the actin cytoskeleton and breakdown of the TJ proteins occludin and ZO-1 (Couraud 1998; Förster et al. 2007). MS is characterised as a chronic and progressive CNS disease with a remitting-relapsing profile. In the course of the disease, both demyelination and microvascular inflammation have been described as central pathophysiological insults. From an animal model for MS, experimental allergic encephalomyelitis (EAE) (Korner and Sedgwick 1996), the inflammatory cytokines TNFα and IL-1 have been delineated to be the key mediators inducing alterations in BBB permeability. Clinically matching data have been collected from MS patients, who show an elevation in TNFα and/or IL-1 in cerebrospinal fluid (CSF) (Weber and Rieckmann 1995). Cytokine-promoted opening of the BBB has been shown to occur due to degradation and decreased synthesis of TJ proteins (Chang and Werb 2001; Harkness et al. 2000; Silwedel and Förster 2006; Yang et al. 2006), resulting in compromised junctional integrity. Particularly, the TJ proteins occludin and ZO-1 have been shown to be negatively affected by the presence of these pro-inflammatory cytokines. Searching for the mechanism of TJ protein degradation, consecutive studies showed, that in many neuroinflammatory conditions, including MS, the matrix metalloproteinases (MMP) play an important role in disrupting the BBB. Amongst these, it has been demonstrated that selectively, MMP-9 (gelatinase B) is increased in CSF levels in MS patients. Tissue inhibitors of metalloproteinases (TIMPs) form complexes with either activated MMPs or with their pro-forms after their secretion, thus regulating MMP activity under physiological conditions (Brew et al. 2000; Yong et al. 2001). Levels of TIMP-1 are however reduced in MS patients relative to control patients, suggesting an imbalance in MMP-9/TIMP-1 ratios in MS (Avolio et al. 2005). MMP-9-mediated opening of the BBB then allows amplification of the inflammation, as demonstrated by radioisotopes (Kermode et al. 1990). Looking for therapeutical agents restoring the unbalaced MMP-9/TIMP-1 ratios in MS, Förster et al. (2007) showed that glucocorticoid treatment reduces the levels of MMP-9 markedly in a cell culture model of the BBB, an effect which was shown to be based on glucocorticoid-mediated transcriptional activation of the TIMP-1 gene. This study further demonstrated that the administration of the glucocorticoids dexamethasone and hydrocortisone preserved the functional integrity of the TJs and adherens junctions under pro-inflammatory conditions, chiefly by maintaining the levels of the TJ components occludin, claudin-1, claudin-12, ZO-1 and VE-cadherin, while dexamethasone effects on claudin-5 were negligible (Blecharz et al. 2008; Förster et al. 2007).
Another study demonstrated differential susceptibility of cerebral and cerebellar endothelial cell lines to BBB breakdown in response to inflammatory stimuli (Silwedel and Förster 2006): increased fragility of the cerebellar BBB had been reported, as the typical caudal to rostral lesion development of MS and EAE with a preference of the cerebellum indicates (Cross et al. 1993; Juhler et al. 1984; Namer et al. 1993). Matching those studies, the authors could demonstrate a faster and more pronounced increase in permeability in the cerebellar BBB manifested by reduced TER and reduced TJ protein expression using the cerebellar BBB model cell line cerebEND (Silwedel and Förster 2006). The stronger effects of TNFα on endothelial cells of the cerebellum as compared the the cerebrum could be explained on the cellular level, where the TJ protein levels of claudin-5 and occludin were shown to be differently affected in cerebellar and cerebral microvascular endothelial cells: effects on occludin were more pronounced in cerebellar endothelial cells, while cerebral endothelial cells of the BBB showed a stronger decrease in claudin-5. Since it was shown that the main protein buiding up the barrier appears to be occludin, while claudin-5 protein, which is also well detectable in non-BBB endothelial cells, does not seem to influence barrier properties to the same extent (Foerster et al. 2002, 2005, 2006; Wolburg and Lippoldt 2002). One may speculate that the lower overall occludin contents of endothelial cells of the cerebellar BBB compared to the cerebral BBB can partly explain the greater sensitivity of the cerebellar BBB to inflammatory stimuli.
Hereditäry diseases
Hereditäry deafness: nonsyndromic recessive deafness DFNB29
The cochlea of the inner ear is divided into two compartments of different ionic composition. Hereby, the perilymph of the scala vestibuli and scala tympani is high in Na+ but low in K+ concentration, while the endolymph of the scala media resembles the intracellular ionic composition, being high in K+ but low in Na+ (Ferrary and Sterkers 1998). This ion gradient leads to the establishment of an 80–100 mV endocochlear potential, necessary for the depolarisation of sensory hair cells, enhancing the sensitivity of mechano-activated channels at the top of stereocilia (Hudspeth 1989).
Besides, hereditary defects in the gap junction proteins connexin-26 and connexin-43 (Sabag et al. 2005) an association between tight junctions and hearing loss was established. The importance of claudin-14 expression in the cochlea was demonstrated by the identification of two different CLDN14 mutations that cause nonsyndromic recessive deafness DFNB29 in two large consanguineous Pakistani families (Wilcox et al. 2001). By in situ hybridization and immunofluorescence labeling mouse claudin-14 expression in the sensory epithelium of the organ of Corti was observed. By the generation and characterisation of claudin-14 knockout mice as a model for autosomal-recessive deafness DFNB29 it was revealed that deafness occurred due to death of outer hair cells. Searching for the factors that promoted increased death of outer hair cells, morphology of the TJs of claudin-14 knockout mice was evaluated. It was demonstrated that TJ strands between supporting cells and between outer hair cells and supporting cells in the organ of Corti are still present and appear morphologically normal in freeze fracture replicas. The analysed TJ strands which are composed of multiple claudins were thus concluded to have rather a functional than an ultrastructural defect (Ben-Yosef et al. 2003). Following the reasoning that claudins create charge-selective channels, Ben-Yosef characterised the electrophysiologic properties of claudin 14 by ectopically expressing it in MDCK cells, which revealed a strong discrimination of transfected monolayers against Na+ and K+ ions. Ben-Yosuf and coworkers thus concluded that the absence of claudin 14 from TJs in the organ of Corti might lead to elevated K+ concentration in the space of Nuel, which is normally filled with the K+-poor cortilymph and increased death of outer hair cells. Long-term exposure of the basolateral membranes of outer hair cells to high K+ concentrations had been shown to be toxic and eventually to lead to cell death before (Zenner 1986).
By the identification of two claudin-14 missense mutations that were unable to form TJs after ectopic expression of CLDN14 in L mouse fibroblastes, Wattenhofer et al. (2005) were then able to prove that the basis for the development of deafness is incorrect recruitment and TJ strand formation in cells expressing mutated CLDN14.
Familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC)
Inherited and acquired defects in epithelial membrane transport are responsible for many human diseases. In the kidney, tubular claudins play a role in extracellular or paracellular permeability. In this context, it has been shown that mutations in claudin-16 (Paracellin-1) impair the function of TJs in the paracellular resorption of Mg2+ and Ca2+ but not of monovalent cations (Simon et al. 1999), probably founding the molecular basis for the development of familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC), an autosomal recessive renal tubular disorder. In FHHNC patients, the impaired tubular reabsorption of Mg2+ and Ca2+ in the thick ascending limb of the loop of Henle with an eventual progression can lead to end-stage renal disease (Lee et al. 2006). Functional tubular reabsorbtion mechanisms are vitally important, since a normal adult absorbs only 200 mg Ca2+ from the diet each day, but loses 8,000 mg to filtration across the glomerulus. To achieve normal calcium balance, the majority of the filtered load must be reabsorbed by the tubule, to approximately 90%, or 7,000 mg/day and this occurs mainly via the paracellular pathway driven by a lumen positive potential across the tubular wall.
The structural backbone of the paracellular pathway must provide for the significant substrate selectivity and functional heterogeneity between nephron segments and thus also plays a pathophysiologic role in renal diseases. Clinically, the symptoms and gradual kidney damage caused by FHHNC cannot be alleviated by intravenous or oral supplementation with Mg2+, and kidney transplantation is often the only option at later stages of the disease.
Recent studies (Simon et al. 1999) have reported that FHHNC is caused by mutations in a gene, which encodes the TJ protein, paracellin-1. Paracellin-1 (PLCN-1) belongs to the claudin family (claudin-16) that allows the paracellular transport of Mg2+ and Ca2+. As a member of the claudin family of TJ proteins claudin-16 exhibits four transmembrane spans with two extracellular loops and both termini located in the cytoplasm. To date, more than 20 distinct mutations have been identified, which affect either the trafficking of PLCN-1/claudin-16, or its permeability properties (Kausalya et al. 2006).
Interestingly, FHHNC patients do not exhibit the salt wasting or metabolic alkalosis that is found in Bartter's syndrome (Wong and Goodenough 1999). Based on these findings, it is tempting to hypothesize that PCLN-1 might form a Mg2+- and Ca2+ -conducting pore. It is therefore likely that PCLN-1 selectively mediates the barrier to divalent but not monovalent cations. Thus, not only can the paracellular barrier discriminate between charged and uncharged solutes, but it may also exhibit an even higher level of specificity distinguishing charge and ionic radius and size of the hydrate shell.
As to molecular investigations based on the use of diverse cell culture models for kidney epithelial cells, there is some discrepancy between results obtained upon overexpression of PLCN-1 in two different kidney epithelial cell lines used. In the canine MDCK cell line, another group found PCLN-1 to decrease Na+ permeability, to decrease TER and to enhance transepithelial Mg2+ transport (Ikari et al. 2006). Another recent study performed in the porcine proximal tubule LLC-PK1 cells suggests that paracellin-1 functions to modulate paracellular conductance and not transcellular transport. They hypothesise that PLCN-1 mediates mostly paracellular Na+ permeation to build up the positive lumen potential and that this lumen-positive potential would be the driving force for the reabsorption of Mg2+ (Hou et al. 2005).
As described for other molecules relevant to Mg2+ homeostasis such as ACDP2, (ancient conserved domain protein), a divalent metal transporter, or the Mg2+ transporter MagT1 (Wong and Goodenough 1999), the human PLCN-1 promoter appears to be responsive to external Mg2+ concentrations, since the renal epithelial cell specific PLCN-1 gene expression was shown to positively correlate with the amount of Mg2+ offered in the growth medium (Efrati et al. 2005).
Further, mutations in PLCN-1 can affect claudin-16 protein targeting at the TJs (Kausalya et al. 2006). The mutation T233R for example prevents interaction of claudin-16 with ZO-1 and results in lysosomal targeting of claudin-16 instead. Protein kinase A (PKA) mediates phosphorylation of claudin-16 at Ser217, failure of which equally leads to dissociation of claudin-16 from ZO-1 and its translocation into the lysosomes (Ikari et al. 2006). In conclusion, although the mechanism of PLCN-1/claudin-16-mediated Mg2+ and Ca2+ reabsorbtion is not yet fully understood, it is clear that these claudins play a crucial role in this process.
Recently, Konrad et al. (2006) additionally unravelled a role for claudin-19 in the development of FHHNC. They had clinically characterized one Swiss and eight Spanish/Hispanic families affected with severe hypomagnesemia due to renal wasting, nephrocalcinosis, and progressive renal failure but had to recognize that the individuals did not have any mutations in CLDN16. They showed that the renal phenotype of the affected patients was virtually undistinguishable from that of patients who presented with FHHNC with proven CLDN16 mutations. However, the affected individuals in these families also have severe visual impairment, characterized by macular colobomata, significant myopia, and horizontal nystagmus (Konrad et al. 2006). Konrad et al. were able to map a new locus for recessive renal magnesium loss on chromosome 1p34.2 and identified mutations in CLDN19, yet another member of the claudin family in patients affected with FHHNC and additionally, with severe ocular abnormalities. CLDN19, which encodes the tight-junction protein claudin-19 was demonstrated to be highy expressed in renal tubules and the retina. In an effort to understand the mechanisms underlying the roles of the two claudins in mediating paracellular ion reabsorption in the kidney, Goodenough and co workers (Hou et al. 2008) showed using pig kidney epithelial cells, that CLDN19 functioned as a Cl(−) blocker, whereas CLDN16 functioned as a Na(+) channel. Mutant forms of CLDN19 were shown to be associated with FHHNC since they were unable to block Cl(−) permeation. Coexpression experiments further elucidated that CLDN16 interacts with CLDN19 and that their association confers cation selectivity, suggesting a role for mutant forms of CLDN16 and CLDN19 in the development of FHHNC.
Vascular system
Diabetic retinopathy
The barrier between the vascular lumen and neural layers in the retina and the brain parenchyme maintains a characteristic microenvironment and is essential for proper neuronal function. This blood-retinal barrier (BRB) consists of two anatomical entities: an inner BRB formed by junctions between endothelial cells of the retinal capillaries, and an outer BRB composed of TJs between retinal pigment epithelial cells. Clinical evidence from fluorescein angiography has demonstrated that the inner BRB appears to be the primary site of vascular leakage leading to macular edema.
The BRB is built up and maintained by TJ complexes that make up the blood vessels of the microvasculature (Mitic and Anderson 1998). The principal proteins found in the retinal endothelial TJs are occludin and claudin-5 (Dejana et al. 2001). Loss of the BRB integrity and vascular permeability leading to macular edema (Vitale et al. 1995) characterise diabetic retinopathy. Macular edema in turn is thought to be directly responsible for vision loss in diabetic retinopathy (Felinski and Antonetti 2005). Vascular endothelial growth factor (VEGF) is thought to be chiefly responsible for this increase in BRB permeability, albeit other permeabilizing factors like histamin or the cytokines IL-1 and TNFα have also been implicated (Felinski and Antonetti 2005). Levels of VEGF, originally identified as an angiongenic factor serving as a mitogen for vascular endothelial cells have been recently correlated with increased vascular permeability (Felinski and Antonetti 2005), compatible with increased VEGF levels detected in the vitreous of diabetic patients for decades. There is enough evidence to show that diabetes and VEGF induce retinal vascular permeability by altering TJ complexes, evidenced for instance by the decrease in occludin content in the retinas of diabetic rats (Felinski and Antonetti 2005). In cultured primary bovine retinal endothelial cells, this observation was mirrored by the observation of decreased occludin contents after VEGF administration (Antonetti et al. 1998). Follow-up investigation underlined this and showed down-regulation of occludin in retinal endothelial cells in culture after VEGF stimulation. VEGF further activates PKC, which leads to increased occludin and ZO-1 phosphorylation (Harhaj et al. 2006). Phosphorylation of occludin and ZO-1 likely contribute to dysregulated endothelial paracellular permeability (Abbott et al. 2006). Regulation of PKC activity and TJ protein modifications may thus have therapeutic implications for treatment of diabetic retinopathy.
Recent investigation has further revealed that alterations of the BRB may involve the active proteolytic breakdown of the endothelial cell TJs by MMPs as also evidenced in the case of MS (compare above) (Leppert et al. 2001). Specifically, elevated levels of MMP-9 have been seen when retinal endothelial cells were exposed to high glucose conditions as a cell culture model of diabetes, leading to proteloytic degradation of specifically the TJ protein occludin followed by disruption of the overall TJ complex (Giebel et al. 2005), rendering the regulation of MMP secretion and activity another therapeutical target in diabetic retinopathy.
Multiple sclerosis
See above.
Tight junction alterations in cancer
Decreased expression of claudin-1 correlates with malignant potential in breast cancer
Changes in TJ function have been shown to be an early and key aspect in cancer metastasis (Ehler et al. 2001). Recent studies showed a role for claudin-1 in invasion and metastasis in mammary glandular differentiation (Förster et al. 2002) and carcinogenesis (Morohashi et al. 2007).
Interestingly, there is an absence or significant loss of claudin-1 expression in several established breast cancer cell lines. Claudin-1 expression in primary human mammary epithelial cells, in contrast to low or undetectable levels of expression in a number of breast tumors and breast cancer cell lines, points to claudin-1 as a possible tumor-suppressor gene. In sections from surgically resected breast specimens, a significant loss of claudin-1 protein in breast cancer cells could be shown by immunohistochemistry (Tokes et al. 2005). This finding suggests that claudin-1 may play a role in invasion and metastasis. The expression profile of claudin-1 in non-malignant versus tumor-derived cells has made this gene an interesting candidate for involvement in tumorigenesis, namely by acting as a suppressor of mammary epithelial proliferation.
In order to evaluate the CLDN-1 gene in sporadic and hereditary breast cancer, Krämer et al. (2000) characterized its genomic organization and screened the four coding exons for somatic mutations in 96 sporadic breast carcinomas and for germline mutations in 93 breast cancer patients with a strong family history of breast cancer. In addition, they compared the 5′-upstream sequences of the human and murine CLDN1 genes to identify putative promoter sequences involved in down-regulation of the CLDN1 gene under malignant conditions and examined both the promoter and coding regions of the human gene in various breast cancer cell lines showing decreased claudin-1 expression. However, neither in sporadic or hereditary breast cancers, nor in breast cancer cell lines they found evidence for a role of aberrant claudin-1 in breast tumorigenesis and concluded that other regulatory or epigenetic factors could be involved in the down-regulation of the CLDN1 gene during breast cancer development.
A correlation between TJ abnormalities and neoplasia can be supported by several observations: alterations in the number, appearance, and permeability of TJs have been demonstrated in various cancer types (Cochand-Priollet et al. 1998; Soler et al. 1999; Swift et al. 1983) and in pre-malignant mammary epithelial cells (Förster et al. 2002). At the molecular level, the Drosophila lethal(1) discs-large-1 gene (dlg) product, which localises to TJ-related insect septate junctions has been shown to regulate epithelial cell proliferation. Genetic loss of dlg leads to a neoplastic overgrowth phenotype (Woods and Bryant 1991). The loss of expression of claudin-1 itself has been demonstrated in several mammary carcinoma cell lines (Swisshelm et al. 1999). The expression of the TJ plaque protein ZO-1 has been shown to be reduced in breast carcinomas or breast cancer cells (Hoover et al. 1998). These findings support the hypothesis that claudin-1 might be involved in the development of breast cancer and possibly in other epithelial tumors too.
The molecular pathways leading toward the loss of claudin-1 expression however, remain to be explained.
Bacterial toxins
Clostridium perfringens enterotoxin
Food poisoning by Clostridium perfringens enterotoxin (CPE) leads to diarrhea symptoms classified as "C. perfringens type A food poisoning and antibiotic-associated diarrhea". The enterotoxin protein has been elucidated to intrude the cells by interacting with epithelial TJ, including certain claudins. Toxin-induced cytolytic pore-formation is a prerequisite for this and involves residues in the N-terminal half of CPE protein, while residues near the C-terminus are required for binding to claudins. In detail, the single 35 kDa CPE polypeptide first causes cellular damage by altering plasma membrane permeability by the formation of an approximately 155 kDa CPE-containing pore complex necessary for the subsequent interaction with TJ proteins via its C-terminal receptor-binding region (McClane 2001). CPE was then perceived to affect TJ structure and function in a way that permeability properties of the epithelial layer are altered leading to diarrhea (McClane 2001).
Further investigation has determined the isoforms of claudins targeted by CPE. Sonoda et al. (1999) were able to show that the COOH-terminal half fragment of CPE (C-CPE) bound effectively to claudin-3 and claudin-4 in MDCK I cells expressing the claudins-1, -2, -3 and -4. They were further able to show that in the presence of C-CPE, reconstituted TJ strands gradually disintegrated and disappeared from their cell surface in C3L cells.
As a consequence, because diverse claudins are overexpressed on some human cancers, the toxin became interesting for targeting chemotherapy in the manner of a Trojan horse (Long et al. 2001; Michl et al. 2001).
Recently, van Itallie et al. (2007) aimed to solve the structure of the CPE claudin-binding domain to advance its therapeutic applications. The structure was shown to be a nine-strand beta sandwich with previously unappreciated similarity to the receptor-binding domains of several other toxins of spore-forming bacteria, giving strong evidence for the presence of a common ancestor for several receptor-binding domains of bacterial toxins (Van Itallie et al. 2007). The very recent elucidation of the structure of a 14-kDa fragment containing residues 194 to the native COOH terminus at position 319 by X-ray diffraction to a resolution below 2 Å showed that the structure is a nine-strand β sandwich with previously unappreciated similarity to the receptor-binding domains of several other toxins of spore-forming bacteria, i.e., the collagen-binding domain of ColG from Clostridium histolyticum and the large Cry family of toxins (including Cry4Ba) of Bacillus thuringiensis (Van Itallie et al. 2008). The authors speculate that the claudin-4 binding site is on a large surface loop between strands β8 and β9 or includes both of these strands. They further were able to clarify that the sequence crystallized binds to purified human claudin-4 with a 1:1 stoichiometry. The binding affinity was determined to be in the submicromolar range similar to that observed for binding of native toxin to cells (Van Itallie et al. 2008). These results could now provide a structural framework to advance therapeutic applications of the toxin.
Conclusion and perspectives
The discovery of the existence of occludin- and claudin-based TJs constituting a permeability barrier in epithelial and endothelial cells has paved the way for the elucidation of the molecular background of diverse human diseases the pathogenesis of which had been unclear before. It became clear that the presence of TJs is indispensable for tissue compartmentalisation and cellular homeostasis. Disturbances in TJ function are reflected in many diseases, but on the other hand, awareness of their involvement has facilitated therapy enormously. For instance, antiinflammatory or antimetastatic drugs could be developed based on their ability to repair broken barrier function. These strategies have become indispensable in the treatment of disease like MS, inflammatory bowel disease or diabetic retinopathy. Following the strategy of pathogenic agents, which use TJ proteins as a docking station to invade cells, even strategies for drug delivery techniques could be implemented. Future attempts will have to address the role of occludin and the exact contribution of the diverse claudins to the formation of ion selective pores, which are still open questions. A more thorough understanding will greatly facilitate diagnosis and the development of specific treatment regimens for diseases originating from impaired TJ function.
This work was supported by grant SFB688/TPA5 from Deutsche Forschungsgemeinschaft DFG to CF. The author is especially thankful toDetlev Drenckhahn for constructive suggestions and critical reading of the manuscript. The author is grateful to Kinga Blecharz for editing expertise and Malgorzata Burek for efficient and pleasant help in collection of material.


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Indiana candidates busy on final full day of campaign
Election Day is so close, and candidates will be out Monday talking to voters one last time.
Election Day is so close, and candidates will be out Monday talking to voters one last time.
The recall follow several reports of injuries — including a broken jaw — due to "excessive vibration."
It happened in McCordsville and Lawrence early Monday morning.
Officials said the majority of the county's snow plows were destroyed.
Reno, a former Miami prosecutor who famously told reporters "I don't do spin," served nearly eight years as attorney general under President…
The earthquake happened just before 7:45 Sunday evening.
The Hershey Company responded in a big way to the candy bar theft.
Crown Hill has become the third northwest side neighborhood to celebrate a year without a homicide.
No injuries have been reported.
The shooting happened just before 7:30 Sunday evening.
A protester involved in a scuffle at a Donald Trump rally in Reno, Nevada denied accusations Sunday that he is an operative for Democrats.
Both are battling to replace retiring Sen. Dan Coats.
The newest Marvel film captured the top spot at the box office.
The announcement comes just days before Election Day.
The shooting happened in Spiceland around 5 a.m. Sunday morning.
The shooting remains under investigation.
It is believed that the victim suffered gunshot wounds to the arm and leg.


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The Victoria and Albert, Natural History and Science museums have written to the Mayor after Transport for London scrapped a scheme to offer step-free access at South Kensington station.
The redevelopment of the station is part of a £30million plan to transform the museum district into a "shared" space for cars and pedestrians.
Initially TfL said it was "likely" a lift would be installed for wheelchair users and parents with prams but it has now said it would cost £50million and there is not enough money.
In their joint letter, the three museum directors - Mark Jones of the V&A, the Science Museum's Professor Chris Rapley, and Dr Michael Dixon at the Natural History Museum - said: "The present situation seems highly regrettable at a time when we need to show our best face to visitors in the run up to the Olympics." About 10 million people visit the museums every year, the vast majority using the Tube.
Today Mr Jones said: "It's an appalling shame that there will be no step-free access. It's inexplicable that this should not happen. We do as much as we can to make our museums accessible and we get people complaining about the Tube. This is a huge lost opportunity."
Merrick Cockell, Kensington and Chelsea council leader, said he thought step-free access was essential for the new-look station.
A TfL spokesman said 54 of all 270 Tube stations had step-free access and that this would rise to 68 stations by 2010 and 71 by 2012.
But he added: "The project to deliver step-free access at South Kensington was part of an unfunded commitment by the previous mayoral administration. London Underground has looked at a number of options including developer-led schemes which have been refused planning consent by the local borough."
The project to upgrade Exhibition Road and its surrounding area will see safety barriers, street signs and traffic lights removed so vehicles share space with pedestrians.
- More about:
- Museums
- Natural History
- Transport For London


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2hp's NSE is a crazy small noise source with VCA mode and continuously variable sample rate. With CV control over sample rate or signal amplitude, this module is an excellent choice for crazy noise sweeps or percussive sounds.
In VCA mode, the trigger input triggers a decay-only envelope that controls the noise's output volume. In this mode, the sample rate knob controls the decay length, allowing the creation of percussive sounds like hi-hats, snares, and more.
- 2hp noise source with controllable sample rate
- Built-in VCA mode for creation of percussive sounds
Questions on NSE Noise Source - 2HP
No questions asked yet
Ask Your Own Question
- Panel width: 2hp
- Max current: +35mA / -16mA
- Depth: 34mm


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Sara Holliday's Prenatal Yoga Series
Fitness guru and yoga instructor Sara Holliday shows expectant moms how to stay in shape, keep stress and fatigue at bay and bond with the baby during nine months of pregnancy. Workouts accommodate physical changes and are divided into trimesters. The first trimester routine tones muscles; the second trimester targets weakened back and legs; and the third trimester prepares women for delivery.
Rent DVDs for only $4.99 a month.
Movie Details
- Cast
- Sara Holliday
- Format
- Ratings
NRNot rated. This movie has not been rated by the MPAA.
Full Screen 1.33:1Subtitles
NoClosed captioned
NoLanguage and sound
English: Dolby Digital 2.0 StereoOther features
Color; interactive menus; scene access.
Sara Holliday's Prenatal Yoga SeriesClose
For Parents
Common Sense Note
- Age appropriate
- Not an issue
- Depends on your kid and your family
- Not appropriate for kids of the age most likely to want to see it


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a novel experienceal
Sail Date: May 10, 2011
Reviewed: 6 years ago
Traveled As: Couple
Room Type: Balcony
Pros: all the conveinces
Cons: the queen size mattress was too soft and also lumpy
Recommends Norwegian Jewel? Yes!
Category Ratings
Click table then swipe to view all columns.
Click table then swipe to view all columns.
|Cabin / Stateroom
was not satisfied with the mattress too lumpy and soft everything else was perfect good size, beautiful view,and clean
|good and had a nice variety
|Ship Quality
ship layout was good everything was accessible and clean
|Food and Dining
|food was excellant there was an abundance of choice and very tasty
|Service and Staff
|every staff member was wonderful to deal with very polite and always willing to aid you whatever you needed
Port Ratings
Click table then swipe to view all columns.
Click table then swipe to view all columns.
|Tortola, British Virgin Islands
|it rained and were not able to see much
|San Juan, Puerto Rico
the galleries are interesting the castles(el morro) are a must the view from there is spectacular the beaches in isla verde are beautiful the artisan in the paseo del prinicesa offer interesting crafts
Don't Miss: should visit el yunque
|Charlotte Amalie, St. Thomas
|st thomas was gorgeous with unbelievable vistas visit the highest point to see a quick overasll view of the city
|Philipsburg, St. Maarten
|the beaches are quite pretty and the ocean is a turquoise blue and clean the shops offer a variety of items and to haggle is welcome


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Birthdays – no matter how old you grow, you never grow out of them! My Dad will turn 66 this year, and me and Mom still buy him a birthday present and make him a nice birthday lunch, and we still go out for dinner and cut a birthday cake, complete with singing 'Happy Birthday To You'! Yes we do it, and you know what? It feels great. Boo the people who think you cannot or should not celebrate your birthday just because you are now old. It is nice to say "It's my life"; but none of us ever really live for ourselves. We don't do everything we do only for ourselves – even the smallest of selfish deeds has a hint of selflessness, if only you look deep enough. But your birthday is YOUR birthday. It is the one single day in your life when everybody wants to talk to you, everybody wants to be with you. It is indeed a special occasion, and it comes around only once a year. So why not click a few pictures and create memories on your special day?
A birthday is the perfect kind of occasion that begs you to click pictures. But it is of no use if you are going to click boring pictures where everybody is simply looking at the camera and smiling mechanically. Photography is about capturing emotions, sentiments, feelings and moments. How do you do that? I am here to tell you exactly that! Read on…
Birthday Party Photography Tips
Children love playing. They are completely devoid of inhibitions and when they are having fun, they are having FUN! They are going to jump, run, play, dance, eat, and OMG! Did that little boy just kiss your girl on her cheeks? LOL. Kids really do all the things that birthday parties are made of. So make sure you are ready with your camera to click as many pictures as you can! You can get some really cute snaps when they are playing games. Their faces are going to be livid with expressions, so make sure you capture those moments. If it is a birthday party with a theme, like say Super-heroes theme for a boy, or Princess theme for a girl, then have something like a spaceship, or a moon crescent cut-out in the garden, or against a plain wall in your house and have the kids pose in front of it to click pictures. (Don't put up a big dinosaur if the theme is Jurassic Park – the kids might just get scared!)
… for a Teenager's Birthday Party
So I am guessing your parents are not going to be around for the entire party right? Use the opportunity! I am sure you have arranged for that cute guy from your class to come over as well, and you must be dying to click pictures with him, right? Make your friends partners in crime, and make sure to get yourself clicked with that guy. If you are going to head outdoors, like to the mall, or for a movie, take your camera along with you. Most cell phones have a camera these days, and a good camera too. So you can even choose to click pictures on your cell phone. Make sure you click pictures with everybody – you and your friends, your friends without you, you and cute guy, you and best friend with your respective cute guys, group pictures, couple pictures, all sorts of combinations. You can even print these pictures and use them in the future, like on a Valentine card maybe…? (Wink Wink!)
… for a 21-Year Old's Birthday Party
Turning 21 spells out lots of things for different people – and one of them is a-l-c-o-h-o-l! You are legally, officially, and (thank God!) finally old enough to get your spirits high! Make sure you click yourself with your first drink in hand, even if it is only wine. If you want some funny or wacky pictures, you could maybe act drunk and click pictures with lots of empty alcohol bottles around! Cheesy? Cheap? So what? It's fun to do stupid things once in a while! Now, 21 could also spell g-a-m-b-l-i-n-g for many of you! So if you are going to hit the local casinos, or better still if you are going to make a trip to Vegas, go click! You first spin at the roulette table, your first hand of Blackjack, even the first time you hit the Jackpot machine is all indeed special, no matter whether your win or lose. Ensure you capture these moments.
… for a 50-Year Old's Birthday Party
Celebrate being a birthday-'boy' or birthday-'girl' again! At 50 I am guessing you are probably married have one or maybe two kids, a nice house and all that jazz. But I am also guessing you have a very hectic job, full of responsibility, a busy life, and no time to just sit down and relax. Why not take the opportunity of your birthday to do something different? You have worked hard to earn the life you are living, now it's time to do something for yourself. Get a make-over. Go on a shopping spree. Spend the afternoon with your gal-pals instead of with the laundry! And click pictures doing all these things. They are things you have not done in a long time, isn't it? Go out on a dinner date with your partner. Or take your partner out if he/she has turned 50!
… for the 60+ Birthday Person
Reminiscence… I guess that is going to be the single most over-powering emotion in your mind as you turn 60, isn't it? Take the opportunity to reconnect with old friends. Social networking websites have made it a lot easy to find almost anybody. Also take the time out to celebrate with your family. Your kids, perhaps your grandchildren as well. Celebrating turning 60 is the perfect kind of occasion to organize a big family reunion. Call everybody – even that cousin you fought with and haven't been speaking to since you were 30. Bury the hatchet, embrace each other, and smile at the camera! Click a huge family picture with everybody in it; and when I say everybody, I mean everybody, including your dog and cat!
Some General Tips
➽ Make sure you click at least two individual pictures – one portrait photograph, and one head-to-toe. Wear your best clothes, turn on your charm, and flash your best smile, for it is your day and you are the star!
➽ Click at least one picture with your Mom and Dad. You may do it after the guests have left or before they arrive, if you feel it is too geeky or out-of-fashion or whatever. But do click one. You will cherish these memories in the later years.
➽ Make sure to click at least one picture with all your friends! Let it be a real group picture, with EVERYONE who came in it. Do not miss out on anybody.
➽ Try and click these 'planned' pictures early on in the party, if it is a house party. Your guests may all leave at different times, so if you leave these picture for the end they may not all be there. Everybody is going to be tired by the end of it, so why look haggard in your own birthday party pictures?
➽ Yes, you need to click a few planned pictures, but that does not mean every picture should have people looking at the camera and smiling. capture some candid moments as well – somebody eating a big piece of cake, somebody having dozed off after the meal, somebody exchanging numbers, and OMG! Did your brother just kiss your best friend on her cheek thinking nobody's watching? Well, he can think again – for you already caught it on camera!
➽ If it is an indoor party, hire a photographer if you can. If that is going beyond your budget, have one family member dedicated to only taking pictures. There are going to be lots of moments to take pictures, you don't want the divided attention of the photographer!
Man invented the arts, so each one of us has got an artistic side to us. It is only a matter of identifying it. Even if you are not into photography, there is a little bit of photography in all of us. Don't we understand when our picture doesn't come out right? If we can understand that, we should be able to figure out how to take a good picture ourselves; it is not that hard. Hope you enjoy your birthday party and click lots of pictures!


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Open Position - Senior Research Assistant/Digital Electronics Technician - CASPER
The Research Assistant/Technician is responsible for supporting research by undergraduate and graduate students as well as faculty from physics, engineering, CASPER, the University of Stuttgart, the Joint Institute for High Temperature (National Academy of Russia), the German Aerospace Center (DLR) and the Wigner Research Institute for Physics (Hungarian National Academy), develops RF pulsed power and digital electronics research equipment, repairs, maintains and operates Class I-IV laser systems, repairs, maintains and operates complex plasma and inductive plasma generator systems, maintains instrumentation and all lab systems in support of the research agenda of the Center. Please see the related job posting for more information.
Questions concerning this position should be directed to the Director of CASPER, Dr. Truell Hyde (firstname.lastname@example.com).


text:
Books Reviewed on www.bibliodish.com
BiblioDish created this book list on IndieBound.org because buying local and independent is important to your economy and your community.
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1) War Dances
Published by: Grove Press
Edition: Paperback (ISBN #9780802144898)
2) Flight
Published by: Grove Press, Black Cat
Edition: Paperback (ISBN #9780802170378)
Published by: City Lights Books
Edition: Paperback (ISBN #9780872865211)
4) The Mirage
Published by: Harper
Edition: Hardcover (ISBN #9780061976223)
Published by: Mariner Books
Edition: Paperback (ISBN #9780156006293)
Published by: Anchor Books
Edition: Paperback (ISBN #9780385720885)
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6 items from 2016
9 August 2016 1:46 PM, PDT | The Playlist | See recent The Playlist news »
"Would you believe that heroism has become old fashioned?" So says a character in Sam Peckinpah's underseen 1975 gem "The Killer Elite." Peckinpah, a man who spent his entire career dealing with the theme of what it means to be a hero, was also known as "Bloody Sam," which gives you all the details you need […]
The post The Nihilistic Cinema Of Sam Peckinpah Is Celebrated In This Career-Spanning Video Montage appeared first on The Playlist. »
- Jordan Ruimy
29 July 2016 11:39 AM, PDT | DailyDead | See recent DailyDead news »
He helmed the 2009 remake of Wes Craven's The Last House on the Left and took viewers to a party they wouldn't soon forget in 2013's +1. Now director Dennis Iliadis is hard at work on his latest film, He's Out There, which has started filming in Canada:
Press Release: Montreal, July 29, 2016 – Principal photography has begun on Screen Gems' suspense thriller, He's Out There. Yvonne Strahovski stars, along with Justin Bruening, Anna Pniowsky, Abigail Pniowsky, Julian Bailey and Ryan McDonald. Dennis Iliadis directs, from a script by Mike Scannell. Producers are Adrienne Biddle and Bryan Bertino of Unbroken Pictures. Glenn Gainor is executive producer.
On vacation at a remote lake house, a mother and her two young daughters must fight for survival after falling into a terrifying and bizarre nightmare.
Yvonne Strahovski's film roles include I, Frankenstein, The Guilt Trip and Killer Elite, and she will next »
- Derek Anderson
14 July 2016 10:00 PM, PDT | avclub.com | See recent The AV Club news »
With A History Of Violence, Tom Breihan picks the most important action movie of every year, starting with the genre's birth and moving right up to whatever Vin Diesel's doing this very minute.
The Octagon (1980)
There's a scene in The Killer Elite, a not-great Sam Peckinpah movie from 1975, where Burt Young, the man who would go on to play Paulie in Rocky a year later, fights a ninja on the deck of a battleship. The ninja does not prove to be much of a problem for him. Young casually picks up the ninja and dumps him overboard. If the ninja even resists, we don't see it. Instead, we see him go screaming into the water. And we see Young—squat, balding, portly, not exactly a physical wonder—leaning on the guardrail and watching him plunge. Then he makes this noise: "Hmp." Like, "That was ...
- Tom Breihan
1 July 2016 1:03 PM, PDT | Dark Horizons | See recent Dark Horizons news »
There's no doubt that Robert De Niro is a great actor, but there's also little argument that his cinematic choices since the turn of the millennium have been rather questionable. As a result, like various acting legends who've reached their elderly years, these days he's been labelled a 'paycheck actor' in various reviews for films like "Dirty Grandpa," "What Just Happened" and "Killer Elite".
Why has this happened to not just him but many other famed acting legends of the 1980s and 1990s? One of De Niro's old co-stars and highly respected actress in her own right, Illeana Douglas, has a theory and says the fault lies less with De Niro and more with the current movie-making environment. Appearing on Bret Easton Ellis' podcast recently, she spoke about her own experiences working with him on films like "Goodfellas" and "Cape Fear":
"I think that, and I can only »
- Garth Franklin
12 March 2016 11:50 AM, PST | Trailers from Hell | See recent Trailers from Hell news »
(This is the first in an occasional series in which I remember some of the best double features I've been lucky enough to see projected in a theater.)
The New Beverly Cinema, the oldest surviving revival theater in Los Angeles, has this week dished up a time-capsule glimpse into America's popular obsession with Cb, or citizen's band, radio and the largely mythological outlaw trucker culture through which it crackled. If you're of a certain age (mine), and you ever cruised around town or down the highway jabbering to friends and strangers on an open channel frequency (I did—my handle was The Godfather!), given the opportunity I don't see how you could possibly resist the chance to see the ultimate trucker-cb action-comedy pairing, Hal Needham's Smokey and the Bandit and Sam Peckinpah's Convoy. (I couldn't!) As of this writing, the morning of »
- Dennis Cozzalio
28 January 2016 3:44 PM, PST | Variety - Film News | See recent Variety - Film News news »
Fresh off the success of "Spotlight," Open Road Films is teaming with Crystal City Entertainment/Boundary Stone Films in a multi-picture development financing pact for all of Open Road Films' own productions over the coming years.
The deal will cover two to four films annually with the titles getting a wide domestic release via Open Road Films and foreign sales being handled by Open Road International and partner Film Nation.
Open Road was launched in 2011 by Regal Entertainment Group and AMC Theaters, two of the largest U.S. exhibitors. Besides "Spotlight," which has topped $33 million domestically and received six Oscar nominations, Open Road has seen strong performances from mid-budget titles such as "The Grey," "Killer Elite," "A Haunted House," "The Host," "Side Effects," "The Nut Job" and "Chef."
Ari Daniel Pinchot of Crystal City Entertainment/Boundary Stone Films said, "Led by Tom Ortenberg, Open Road Films has a fantastic track »
- Dave McNary
6 items from 2016
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Three-bedroom apartment with one bathroom, separate kitchen, terrace, optional garage and storage. the apartment is at 50m de la calle campos
We can help you to get a mortgage on favourable terms. A residence permit can be obtained with the purchase of this property. Please ask our team for more information.


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Phones are not made for swimming – or dropping into toilets – but sometimes the unfortunate happens and we're left with a wet dead phone and a lot of anguish!
(And it doesn't make it any more pleasant that the most common place to drop them is in the toilet!)
Recently, a friend and colleague of mine, Jim, posted to Facebook…
"Ok, my FB friends, I need YOUR HELP! I dropped my old phone into a puddle of water. "
Sounds like game-over for the phone right? He was pretty sure he was going to need to buy a new (and expensive) replacement.
I quickly sent him a reply…
"And I assume you're ready to switch? Because if you're not, then you need to immediately pull the battery out of the wet phone. Towel it off. Let it air dry a little bit near a fan. Then get it into a dessicant, such as dry white rice, to let it pull the moisture out from inside, and let it stay there a minimum of 24 hrs (longer the better) before you try powering it on."
First for a little inappropriate laugh to help break the stress…
Now, lets get into some of the actual details of what you need to do!
This video from Pablo Muñoz was a great walk-through of the steps.
A couple tips:
(1) Pull the battery out immediately. The minute you get the phone out of the water, get the battery out. That battery is the part that can ruin the board in an instant.
(2) You must use a CLOSED container for this even if that means you need to use a large ziploc-style baggie instead. You want the rice to pull the moisture from inside the circuit board rather than from the air in the room.
(3) The hard part is being patient and not opening it two dozen times and letting it stay long enough to get dry inside in the parts you can't see. It's a MINIMUM of 24 hour dry time and better to go longer. Each time you open the container, you add time. If you get to the time, pull the phone out, assemble it, and try to power it on. If it comes up, you're probably good. If it doesn't, sometimes another 24hrs can make a difference and at this point you have nothing to lose by trying it.
The goal is to get the internal board 100% dry – rather than "just dry enough to turn on". If it's "just dry enough to turn on" then the heat of the phone will dry it out – but at the cost of serious corrosion to the board which will eventually result in a malfunction even if it's not today.
Yeah, but does it work?
It sure does!
Now, I suggest bookmarking this post so that you have the information handy if you ever (unfortunately need it).
(If you give the post a +1 on G+, and are logged into Google when you search, the post will come up for you as well.)
It's a really remarkable trick that can save a LOT of devices – but it must be done soon after the "swimming lesson"!
Good luck!
~ Kim ~
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