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Clean water can save a newborn baby's life; let's commit to giving them a #HealthyStart
Today is World Water Day which is an important annual milestone for international development. Providing universal access to clean drinking…
We are still fighting against racial discrimination in the EU
On this day in 1960, South African police opened fire against 69 people demonstrating peacefully against apartheid 'pass laws' in…
30 years on from the first .com domain, the internet has transformed our lives
Today marks 30 years since the first .com domain name was registered. Back in 1985, American computer manufacturer Symbolics acquired…
Bringing back high-quality apprenticeships is vital for our young people
This week marks National Apprenticeship Week which is a fantastic opportunity to bang the drum for apprentices and the huge…
Labour MEPs vote for credit card fees cap to help small businesses
Labour MEPs have voted in favour of capping charges for credit and debit card transactions which will save British businesses…
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Local MEP calls for urgent action on tuberculosis on World TB Day
A Local Euro MP is calling for concerted action to deal with the threat of multi-drug resistant tuberculosis (MDR-TB). Glenis Willmott is urging EU Member States to make tackling MDR-TB a priority and to work together to ensure that everyone has access to affordable and effective treatments for this infectious disease.
24th March marks World TB Day and new figures released this week suggest 75 million people could lose their lives to TB over the next 35 years if action is not taken to deal with MDR-TB.
Glenis Willmott MEP, Labour's European Health Spokesperson, said:
"Many people think of TB as a disease of the past but sadly this is far from the case. Globally TB still kills 1.5 million people every year and 9 million news cases occur annually; one in twelve new TB diagnoses in Europe are MDR-TB.
"I am deeply shocked by the figures released this week showing that the number of deaths as a result of TB could increase significantly if we do not take urgent action to counter the growing threat of drug-resistance."
It is estimated that by 2050, TB could cost the global economy $16.7 trillion, yet development of new treatments has been slow, with only two new anti-TB drugs approved by the European Medicines Agency in the last 20 years. Resistance often develops because patients do not have access to an adequate supply of drugs or are given poor quality medication.
Latvia, which currently holds the Presidency of the EU, will host a ministerial meeting to address TB and MDR-TB in Riga next week and earlier this year the Global TB Caucus launched the Barcelona Declaration, to show the political commitment of parliamentarians from across the world to make fighting TB a priority.
Glenis Willmott said:
"As chair of the European Parliament's Access to Medicines Working Group, I know that one of the major hurdles to dealing with TB is the fact that many patients simply do not have access to timely diagnosis and affordable and effective treatments.
"We urgently need coordinated action from governments, the EU and international organisations such as the World Health Organisation. I have signed the Barcelona Declaration and hope that when EU Health Ministers meet in Riga next week, they will agree concrete action to make dealing with TB and MDR-TB a priority.
"We all need to work together to ensure that one day TB really will be a disease of the past."
Clean water can save a newborn baby's life; let's commit to giving them a #HealthyStart
Today is World Water Day which is an important annual milestone for international development. Providing universal access to clean drinking water has long been something I have campaigned on and a key goal of the United Nations and countless brilliant charities including WaterAid, charity: water, Just a Drop and many more. But with the UN Millennium Development Goals coming to an end this year, we need to redouble our efforts to help the millions of people still in desperate need of this basic yet vital resource that we take for granted.
Although we have made fantastic progress since many of those charities were started, today's figures are still shocking: a child dies of a waterborne disease every 20 seconds, that is more than 1.5 million a year, and 750 million people lack access to safe, clean drinking water. It costs just £15 to provide one person with access to clean water; not a high price to save a life.
But the issues surrounding clean water don't just affect drinking; more than 2.5 billion people, over a third of the world's population, do not have access to adequate sanitation. This has enormous consequences for hygiene in some of the world's poorest areas, but it can also put women and young girls in danger when they are forced to be outside alone.
Perhaps the most terrible statistic of all is the one in five babies in developing countries who do not make it past their first month; all for the want of clean water to wash them and their carers' hands. This year, WaterAid is launching the #HealthyStart campaign which is calling for universal access to water and sanitation by 2030 to be one of the UN's Sustainable Development Goals currently being drawn up. The human cost of failing to invest in quality, robust healthcare in the developing world is clear to see which is why we must take the opportunities that this year offers us to push for real progress.
Many of the current Millennium Development Goals are significantly affected by access to clean water: reducing child mortality, improving maternal health, promoting gender equality, to name a few. This is why it is so important that water, sanitation and hygiene are at the forefront of our minds as the post-2015 UN development framework is planned and the 2015 European Year for Development is taking place in the EU.
The EU has played a significant role in delivering safe water and sanitation with around £300 million being committed every year since 2004 through projects such as the European Water Initiative and the EU-Africa Partnership on Infrastructure. EU projects recognise that water has the potential to not only unlock better hygiene and health, but also better nutrition, security and energy generation.
I look forward to continuing my work with colleagues in European Parliament to ensure that 2015 is the year we begin the end of these shocking statistics and give today's babies a healthy start in life.
We are still fighting against racial discrimination in the EU
On this day in 1960, South African police opened fire against 69 people demonstrating peacefully against apartheid 'pass laws' in Sharpeville, South Africa. The United Nations International Day for the Elimination of Racial Discrimination marks this tragedy every year, and it should serve as a reminder of the evils that prejudice can cause.
It was a Labour government that passed both the Race Relations Act in 1976 and the Equality Act in 2010, and we should be glad to live in a society where it is illegal to discriminate on the basis of race in employment and access to public or private services.
While it is undeniable that we have made great progress in the last few decades, Nigel Farage's suggestion that we no longer need anti-discrimination laws in employment is foolish.
New figures this month have shown us that the number of young people from black, asian and minority ethnic (BAME) backgrounds out of work for more than a year has risen by almost 50% under the Tory – Lib Dem coalition. This isn't just a problem for those young people, but for our society as a whole. By stopping a generation of BAME young people from fulfilling their potential, we are limiting the number of talented individuals who might enter the judiciary, the boardroom or even politics in the future.
A Labour government would not stand back while talented young people are struggling to find employment, and we certainly would not scrap anti-discrimination laws. Labour will guarantee a real, paid, starter job to every 18 to 24 year old who has been claiming Jobseeker's Allowance for more than a year.
We should remember that racial discrimination and disparities in employment aren't just issues in the UK, but EU wide, and more should be done at EU level to address the problem.
We have recently seen the worrying rise of far right groups in the European Parliament, giving their MEPs a platform to spread bigotry. The comments of some of the far right MEPs are shocking and we need to make it clear to these groups that the European Union is based on values of tolerance and respect.
It is incredibly important for us to stand up and defend these European values – failure to do so risks normalising the sort of discrimination that we should have left behind many years ago. Labour MEPs have encountered appalling views in the European Parliament, and we will always challenge those who hold them.
Labour MEP supports campaign for World Down Syndrome Day
A local Euro MP is encouraging everyone to help raise awareness of Down syndrome.
Glenis Willmott, Labour MEP for the East Midlands, is supporting the UN's World Down Syndrome Day this Saturday, 21st March. Labour MEPs are encouraging everyone to help raise awareness, and prompt people to learn more about the vital roles that people with the condition play in our communities.
Glenis Willmott MEP said:
"People with Down syndrome should be able to enjoy equal opportunities as children, and have equal choices about how they live their lives as adults.
"Tomorrow is about celebrating the achievements of people with Down syndrome, and demonstrating how, with support, they can play an active role in our communities."
Approximately 60,000 people in the UK have Down syndrome, which occurs in almost 1 in every 1000 births. Improved understanding by health workers and the general public has opened up new opportunities for people with the condition, enabling them to lead active, healthy lives.
Glenis Willmott MEP added:
"A lot of people don't really know what it means to have Down syndrome, so Labour MEPs want to encourage people to take this opportunity to find out more."
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By Michael S. Fischer |
June 24, 2009
Kevin Ellman, chief executive of Wealth Preservation Solutions in Ridgewood, N.J., began using exchange-traded funds in its portfolios some 10 years ago, after he had become disenchanted with active money managers.
By Stephen Phillip Brown |
June 1, 2007
Consider the mind-boggling possibilities of compound interest: If the Lenape tribe of Native Americans had invested the 60-guilder ($24) proceeds from their sale of Manhattan...
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Introducing an Advanced Investment Product Advisor Course at CFFP & a presentation focusing on financial planning, expectations and strategies for different generations.
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Seventy percent of consumers over the age of 65 will need long term care (LTC) at some point in their lifetime. In this 60-minute...
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Comment: Easier said than done (Score 1) 330
by mac1235 (#45447139) Attached to: Can the US Be Weaned Off Ethanol?
Good idea. Go do that.
We've improved Slashdot's video section; now you can view our video interviews, product close-ups and site visits with all the usual Slashdot options to comment, share, etc. No more walled garden! It's a work in progress -- we hope you'll check it out (Learn more about the recent updates).
I thought C++, (as a well established Programming language) would have more libraries than XNA, which I had not previously heard of?
CCI Power 6/40: one board, a megabyte of cache, and an attitude...


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Google Asks Government For More Transparency, Other Groups Push Back Against NSA 323
from the do-not-go-gentle dept.
We've improved Slashdot's video section; now you can view our video interviews, product close-ups and site visits with all the usual Slashdot options to comment, share, etc. No more walled garden! It's a work in progress -- we hope you'll check it out (Learn more about the recent updates).
"Anonymous Coward" has already signed that petition. Maybe you should get a real name if you want to show support?
If the FISA courts have never denied a request, then that's proof nothing improper's going on!
Nobody's even asking for anything out of line. The system is working.
"Mr. Watson, come here, I want you." -- Alexander Graham Bell


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Rebecca Lieb | October 28, 2008 | Comments
Recession got you down? Been laid off or downsized? One man, Joel Moss Levinson, has earned over $200,000 in cash and prizes, becoming the king of CGM in the process
Recession got you down? Been laid off or downsized?
One man, Joel Moss Levinson, has earned over $200,000 in cash and prizes, becoming the king of CGM in the process. Today's NY Times profile describes, and links to, campaigns Levinson has created videos for brands as diverse as Best Western, Klondike, Little Penguin wine, Delta, and the American National CattleWomen, to name just a few.
His contest-winning is due in no small part to a social media marketing strategy, asking his Facebook and Twitter friends and followers to vote for his entries.
Hey, we all know CGM's big. But who knew it can be a living? Maybe you don't need that new agency gig after all.
Want to learn more?
Attend ClickZ Live New York March 30 - April 1. With over 15 years' experience delivering industry-leading events, ClickZ Live brings together over 60 expert speakers to offer an action-packed, educationally-focused agenda covering all aspects of digital marketing. Register today!
Rebecca was previously VP, U.S. operations of Econsultancy, an independent source of advice and insight on digital marketing and e-commerce. Earlier, she held executive marketing and communications positions at strategic e-services companies, including Siegel & Gale, and has worked in the same capacity for global entertainment and media companies, including Universal Television & Networks Group (formerly USA Networks International) and Bertelsmann's RTL Television. As a journalist, she's written on media for numerous publications, including "The New York Times" and "The Wall Street Journal." Rebecca spent five years as Variety's Berlin-based German/Eastern European bureau chief. Rebecca also taught at New York University's Center for Publishing, where she also served on the Electronic Publishing Advisory Group. Rebecca, author of "The Truth About Search Engine Optimization," was ClickZ's editor-in-chief for over seven years.
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Mo. officers fatally shoot knife-wielding man
The shooting is currently under investigation and may have been a 'suicide by cop.'
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- 3 Wis. trooper, suspect, motorist killed in shootings
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- 5 Suspect dead, 8 cops injured in Ariz. melee


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Events for March 25, 2013
The Path of Nature: French Paintings from the Wheelock Whitney Collection
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Guitar World presents five headphone songs guaranteed to blow your mind.
These rockers weren't called "hair" bands for nothing!
Enter to win a gorgeous new acoustic guitar from D'Angelico and some inspiring new music played on a D'Angelico guitar from Will Dailey!
The Eagles' 1976 mega-hit single "Hotel California" turns up — time and time again — on lists of "best guitar solos of all time." In fact, it earned the No. 8 spot on Guitar World's often-quoted list of the 100 greatest guitar solos.
Maybe The Rolling Stones should kick off their two November shows at London's O2 Arena with "All Sold Out," a track from Between the Buttons.
Check out this newly posted (and very funny) video of Gary Rossington and the rest of Lynyrd Skynyrd bemoaning their hillbilly associations. The video is a teaser for an episode of E!'s The Soup, which will air 10:30 p.m. EST (9:30 Central) this Sunday. The episode is called The Goodest Moments Of Redneck Shows.
Led Zeppelin's new live film, Celebration Day, hit theaters earlier this week. If you haven't been enticed into checking it out just yet, the band have posted yet another teaser clip from the movie, this one featuring several crowd shots and an all-too-brief listen at a live version of "Whole Lotta Love." Check it out below.
Cradle of Filth won't release their new album, The Manticore and Other Horrors, until October 30, but we're teaming up with those masters of darkness today to bring you the exclusive North American premiere of their new music video, "Frost on Her Pillow." Check it out below.
Check out this exclusive premiere of "Here Next To Murderous," a new track by Norwegian quintet Blood Command. The song is from their upcoming album, Funeral Beach, which will be released stateside on November 13 via Fysisk Format. It's the followup to 2010's Ghostclocks.
Aerosmith have just released their second official video from their new album, Music From Another Dimension!. Check out the clip for "What Could Have Been Love" below.
Guitar Legends: Classic Rock takes you back in time to late Sixties, the Seventies and the Eighties. In this one jam-packed issue, you get a look at the greatest bands that shaped rock and roll, including Led Zeppelin, Pink Floyd, Van Halen, AC/DC and many others.
BOSS is hosting the 3rd Annual BOSS Loop Station U.S. Finals, which will be broadcast live from Musicians Institute in Hollywood, California, on GuitarTV.com. A roster of musicians and music industry leaders will crown the best looper in America as contestants compete for their spot on the world stage of looping.
Cort Guitars has announced the second signature guitar from Scorpions guitarist Matthias Jabs — the Arena 1. The guitar features Scorpu pickups, which were designed by Jabs and made to his exact humbucker/single coil/humbucker specs.
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DEFINITION of 'Composite Index Of Lagging Indicators'
An index published monthly by the Conference Board that is used to confirm the direction of the economy's movements in past months. The index is made up of the following seven economic components, whose changes tend to come after changes in the overall economy:
1. The value of outstanding commercial and industrial loans
2. The change in the Consumer Price Index for services from the previous month
3. The change in labor cost per unit of labor output
4. The ratio of manufacturing and trade inventories to sales made
5. The ratio of consumer credit outstanding to personal income
6. The average prime rate charged by banks
7. The inverted average length of employment
INVESTOPEDIA EXPLAINS 'Composite Index Of Lagging Indicators'
As it measures the economic activities of previous months, the Composite Index of Lagging Indicators is used as an after-the-fact way to help confirm economists' assessments of current economic conditions. For this purpose, the Composite Index of Lagging Indicators is best used in conjunction with the Composite Index of Coincident Indicators and Composite Index of Leading Indicators.
Business CycleThe fluctuations in economic activity that an economy experiences ...
Consumer Price Index - CPIA measure that examines the weighted average of prices of a basket ...
Business Cycle Indicators - BCIComposite of leading, lagging and coincident indexes created ...
Lagging Indicator1. A measurable economic factor that changes after the economy ...
Central BankThe entity responsible for overseeing the monetary system for ...
Consumer Confidence Index - CCIA survey by the Conference Board that measures how optimistic ...
What are leading, lagging and coincident indicators? What are they for?An indicator is anything that can be used to predict future financial or economic trends. For example, the social and economic ... Read Full Answer >>
A Guide To Conference Board IndicatorsLearn to put the CB data sets to trading use. Each chapter takes you through one of the board's benchmark indicators or surveys, their significance and their applications.
Economic Indicators To KnowThe economy has a large impact on the market. Learn how to interpret the most important reports.
Credit & Loans
Top US Housing Market IndicatorsA quick overview of the top economic indicators to track the housing market in the US.
What's Driving U.S. Stocks? Irony.A seesaw week for U.S. stocks ended on the upside last week, though the rally was more a function of slow growth rather than a booming economy.
Stock Analysis
A Trader's Look at the S&P 500Moving averages are sending an important signal, and it's probably not what you think.
Credit & Loans
The Tangled Web of Interest Rates, Mortgage Rates, And The EconomyThe article looks into the interwoven relationship between the economy, interest rates, and mortgage rates.
Mutual Funds & ETFs
Top ETFs Tracking the U.S. EconomyBullish about the U.S. Economy? Consider these ETFs.
What's Missing For A Turnaround In Brazil?Today, many investors have written off Brazil following several years of poor performance, decelerating growth and chronically high inflation.
Worried About The Fed Hike?With the Nasdaq crossing the threshold and the broader equity bull market entering its 7th year, many investors are anxious that stocks are in a bubble.
Could Ukraine's Debt Crisis Destabilize The EU?A financial crisis in Ukraine could be a bigger threat to European security than the current financial crisis in Greece.


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Archived Story
Niles woman waits 15 years for first pecanPublished 5:04pm Thursday, August 23, 2012
When growing pecans in Michigan, the biggest key is patience.
Just ask 83-year-old Irene Leamon, who waited 15 years to see one of the six pecan trees on her Dick Street property bear fruit.
The Niles native saw the first cluster of pecans, still in their outer husks, last week during a rainstorm.
"I thought, 'oh, I've got pecans,'" Leamon said. "When it quit raining I went out and looked and sure enough there were nuts there."
Leamon started growing her first pecan tree from seed some 15 years ago.
She and her husband, Robert, got the seed in Oklahoma while making one of their yearly visits to Irene's lifelong friend Doris Monks. Both graduated from Niles High School in 1946 and both shared a deep love for the pecan.
"It's our favorite nut," Irene said.
When pecans are ripe, Irene said, the outer shell cracks open and the pecan falls to the ground. While in Oklahoma, Irene and Monks spent most mornings picking up pecans from the ground.
"We'd bring home enough pecans to eat for a year," Irene said.
When Irene decided to grow her own pecan tree in Michigan, she said most people told her it couldn't be done in this climate.
Irene tried anyway, and with the aid of Monks' growing advice, the tree finally produced fruit this year.
All that remains is waiting for the nut to ripen.
"I'm just glad the good Lord let me live long enough to see a pecan on my own tree," Irene said.
Unfortunately, Irene couldn't share the moment with Monks, who passed away around three years ago.
"She'd have been tickled to see it," Irene said. "Every time I see a tree I can think of Doris and how we used to go up and down the street picking pecans."


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|Table of Contents
|Vol. 115, No. 2, 2010
Issue release date: June 2010
|Section title: Original Paper
Albumin Uptake in OK Cells Exposed to Rotenone: A Model for Studying the Effects of Mitochondrial Dysfunction on Endocytosis in the Proximal Tubule?Hall A.M.a, b · Campanella M.b, d · Loesch A.c · Duchen M.R.b · Unwin R.J.a
aCentre for Nephrology, bDepartment of Cell and Developmental Biology, and cCentre for Inflammation, UCL, and dRoyal Veterinary College, University of London,London, UK Corresponding Author
Background: The renal proximal tubule (PT) is clinically vulnerable to mitochondrial dysfunction; sub-lethal injury can lead to the Fanconi syndrome, with elevated urinary excretion of low-molecular-weight proteins. As the mechanism that couples mitochondrial dysfunction to impaired PT low-molecular weight protein uptake is unknown, we investigated the effect of respiratory chain (RC) inhibitors on endocytosis of FITC-albumin in PT-derived OK cells. Methods: Uptake of FITC-albumin was quantified using confocal microscopy. Cytosolic ATP levels were measured in real time using both luciferin/luciferase assays and measurements of free [Mg2+]. Reactive oxygen species production was measured using mitosox. Results: RC blockade produced only a small decrease in cytosolic ATP levels and had minimal effect on FITC-albumin uptake. Inhibition of glycolysis caused a much bigger decrease in both cytosolic ATP levels and FITC-albumin endocytosis. Rotenone led to higher rates of reactive oxygen species production than other RC inhibitors. Rotenone also caused widespread structural damage on electron microscopy, which was mimicked by colchicine and prevented by taxol; consistent with inhibition of microtubule polymerisation as the underlying mechanism. Conclusions: Endocytosis of FITC-albumin is ATP-dependent in OK cells, but the cells are very glycolytic and therefore represent a poor metabolic model of the PT. Rotenone has toxic extra-mitochondrial structural effects.
© 2010 S. Karger AG, Basel
The renal proximal tubule (PT) is responsible for the uptake of filtered low-molecular-weight proteins (LMWPs) and peptides [1] such as retinol-binding protein, parathyroid hormone and insulin, which have to be reclaimed to prevent their loss in urine. These LMWPs are thought to be scavenged from the luminal fluid by a low-affinity, high-capacity, uptake system, consisting of 2 large receptors situated in the apical membrane, megalin and cubilin [2]. On ligand binding, the receptor-ligand complexes are internalised, via receptor-mediated endocytosis, in endosomes, which are then acidified to promote dissociation of the ligand from the receptors [3], and the receptors recycled to the luminal membrane.
The large amount of solute transport that occurs in the PT requires energy in the form of ATP that is generated largely by mitochondrial oxidative phosphorylation, which produces more ATP per substrate molecule than anaerobic glycolysis [4]. Clinically the PT is vulnerable to mitochondrial dysfunction from insults such as genetic mitochondrial cytopathy [5], toxic xenobiotics (e.g. anti-retroviral therapy [6,7]) and ischaemia-reperfusion injury [8]. The reasons for this are unclear, but it has been demonstrated previously in isolated tubules that the PT has very little capacity for anaerobic metabolism [9], and we have shown recently – using multi-photon imaging of rat kidney slices – that mitochondrial function and the response to respiratory chain (RC) inhibition varies along the nephron [10]. Sub-lethal injury in the PT can cause a breakdown in normal transport processes leading to urinary losses of solutes, including LMWPs (the renal Fanconi syndrome [11]). The pathogenesis of the Fanconi syndrome is poorly understood, but a defect in cellular metabolism could be a unifying mechanism underlying its various aetiologies [12].
The exact mechanism(s) by which mitochondrial dysfunction can cause Fanconi syndrome and impaired LMWP uptake is unknown, although a number of steps in the endocytic pathway are thought to be ATP dependent. However, given the varied and important cellular roles of mitochondria apart from ATP synthesis [13], it is quite possible that abnormalities in mitochondrial function could affect endocytosis by non-ATP-mediated mechanisms. For example, mitochondria are significant generators of intracellular reactive oxygen species (ROS; mainly at RC complexes I and III [14]), which have been shown to be involved in endocytosis of membrane Na/K-ATPase in alveolar epithelial cells [15], and shear stress-induced fluid-phase endocytosis in vascular endothelial cells [16].
Opossum kidney (OK) cells are an immortalised PT cell line – derived from the American opossum (Didelphys virginiana) – that form a polarised monolayer with tight junctions and a brush border [17]. OK cells have been used extensively as an in vitro model of the PT epithelium, and they take up LMWPs via receptor-mediated endocytosis [18]; indeed, the rate of LMWP uptake is thought to be greater than in other PT-derived cell lines. The expression of megalin and cubilin in OK cells has been shown previously [19]. Although not technically a LMWP, albumin is typically used for protein uptake experiments in OK cells, as it is endocytosed avidly, and it is an established ligand for the megalin-cubilin system [19]; uptake of albumin is thought to occur in the PT in vivo [20]. OK cells have also been used to study the uptake of dextrans by fluid-phase endocytosis [3], which may also contribute to LMWP uptake in the PT. Rotenone is a well-established inhibitor of complex I of the mitochondrial RC, and it is widely used in metabolic research.
By studying the effects of RC inhibitors (including rotenone) on albumin endocytosis, on ATP levels and on ROS production in OK cells in vitro, we wanted to investigate how mitochondrial dysfunction might impair LMWP uptake in the PT.
Magnesium green, cytosolically targeted luciferase and mitosox were all acquired from Molecular Probes (Invitrogen, Paisley, UK). All other reagents were obtained from Sigma-Aldrich (Poole, Dorset, UK), unless otherwise stated.
OK cells were kindly provided by Dr. Alex Pearson (South West Thames Institute for Renal Research, Carshalton, UK). They were grown in DMEM:F12 supplemented with 5% fetal bovine serum, but without antibiotics, and passaged every 6 days. For experiments, 13-mm-diameter sterile coverslips were coated with poly-L-lysine for 10 min (to assist cell adhesion); this was then washed off with sterile water and the coverslips were allowed to dry. Cells were seeded onto the coverslips in 24-well plates and grown until confluent (5–6 days).
To establish whether or not OK cells were alive after exposure to RC inhibitors for 2 h, cells were loaded with Hoechst (5 µM – excited at 720 nm) and propidium iodide (10 µM – excited at 514 nm). The former stains all cell nuclei, whereas the latter is normally excluded by an intact cell membrane, and stains the nuclei of permeablised and dead cells [21]. When assessing the specific toxicity of the glycolytic inhibitor 2-deoxyglucose (2-DG), pyruvate (5 mM) was added to the solution as a substrate for oxidative phosphorylation.
FITC-labelled albumin (FA) was used as a ligand for receptor-mediated endocytosis by OK cells, as they are known to endocytose it avidly [3]. FITC-labelled dextran (FD) was used to investigate fluid-phase endocytosis (pinocytosis). For endocytosis experiments, OK cells were washed 4× in a HEPES-buffered solution (pH 7.4) containing (in millimol/litre): NaCl 138, KCl 5.6, NaHCO3 4.2, NaH2PO4 1.2, MgCl2 1.2, CaCl2 2.6, glucose 10 and HEPES 10. They were then incubated in HEPES-buffered solution (pH 7.4) containing FA (50 µg/ml) or FD (5 mg/ml) for 1 h at 37°C. The cells were then washed with 0.2 M acetic acid/0.5 M NaCl to remove any FA bound to the external cell membrane, and then subsequently 6× with ice-cold HEPES-buffered solution to remove any FA or FD in solution and to prevent further endocytosis (which is temperature-dependent). The cells were fixed with 4% para-formaldehyde for 15 min and mounted on slides with Citifluor AF1 glycerol/phosphate-buffered saline mountant (Citifluor Ltd., London, UK), and imaged with a Zeiss LSM 510 inverted confocal microscope (Carl Zeiss Ltd., Welwyn Garden City, Hertfordshire, UK). Random fields were selected using the bright field setting and a ×40 oil immersion objective; a 10-µm-deep image z-stack (with 1-µm intervals) was imaged for each field at 488 nm. The mean fluorescence signal collected for each stack was calculated with Zeiss LSM software with identical threshold values throughout.
The polarity of endocytosis in OK cells (i.e. apical versus basolateral uptake) was established by growing cells to confluence on Transwell plates (Appleton Woods, Birmingham, UK). FA was then applied to either the apical or basal side of the insert using the protocol above. Inserts were removed and mounted with Mowiol (a mountant that sets solid – a kind gift from Dr N. Carvou, UCL, UK) and imaged according to the same protocol as above.
To investigate the effects of mitochondrial RC inhibitors on the uptake of FA or FD in OK cells, rotenone (10 µM), piericidin (10 µM) or cyanide (1 mM) (all at supra-maximal concentrations) were added to the HEPES-buffered solution at the start of the 1-hour incubation to inhibit complex I (rotenone or piericidin) or IV (cyanide) of the RC; 5 mM of 2-DG was used (in the presence of 5 mM of pyruvate) to investigate the effect of inhibiting glycolysis.
Two different methods were used to measure the effects of RC inhibition on cytosolic ATP levels in real time. The first involved the use of the fluorescent indicators magnesium green and tetramethyl rhodamine methyl ester (TMRM) to provide a dual readout of changes in ATP levels and mitochondrial membrane potential (Δψm) [22]. The principle of this measurement is that cytosolic free [Mg2+] rises as a consequence of ATP depletion. This occurs because ATP has a higher affinity for Mg2+ than ADP, and most Mg2+ in the cell is bound to ATP. Magnesium green fluorescence signal intensity changes in proportion to increases or decreases in [Mg2+] [22]; therefore, the fluorescence signal changes inversely to changes in [ATP]. Meanwhile, alterations in the mitochondrial retention of TMRM represent changes in Δψm. Magnesium green was loaded at a concentration of 5 µM with 0.01% pluronic (and 2.5 mM of probenecid to prevent dye extrusion by organic anion transporters [23]) at 20°C for 20 min. It was then washed off and the cells were incubated for a further 20 min with TMRM 50 nM (and verapamil 100 µM to prevent efflux of the drug by multidrug resistance transporters [24]). At this low concentration TMRM operates in the 'linear range' (i.e. fluorescence signal is proportional to dye concentration), and dye concentration reflects the Nernstian distribution between compartments according to local potential differences. Therefore, a reduction in Δψm results in a decrease in mitochondrial specific TMRM fluorescence signal and a corresponding increase in cytosolic TMRM signal [25]. Magnesium green was excited at 488 nm and TMRM at 543 nm; light was collected using 505- to 530-nm band-pass and 560-nm long-pass filters, respectively. Much of the extra-nuclear volume of OK cells is filled with mitochondria; to obtain a 'cytosolic' signal for recording simultaneous changes in magnesium green and TMRM, regions of interest (ROIs) within the nuclei of cells were used for analysis.
The second method applied for measuring changes in ATP levels was a luciferin-luciferase assay; alterations in luminescence were recorded using a custom-built luminometer (Cairn Research, Faversham, Kent, UK). OK cells were tansfected with luciferase targeted to the cytosol. Cells were perfused with HEPES-buffered solution and supplemented with 50 µM of luciferin (Promega Ltd., Southampton, UK). For both ATP assay methods the RC inhibitor rotenone (10 µM) and the glycolysis inhibitor iodoacetic acid (IAA – 1 mM) were added sequentially to investigate their effects.
To measure mitochondrial ROS production, OK cells were loaded with the superoxide-sensitive dye mitosox (5 µM), which localizes to mitochondria. The rate of increase in fluorescence signal is proportional to the rate of ROS production. The cells were incubated with the dye for 10 min at room temperature before imaging. Mitosox was excited at 543 nm and light was collected using a 560- to 615-nm band-pass filter. Images were acquired every 2 min to minimize laser-induced photo-toxicity. After a 10-min control period, the RC inhibitors rotenone (10 µM), piericidin (10 µM) or cyanide (1 mM) were added. ROIs were drawn around areas of cells with high mitochondrial density.
To perform electron microscopy, OK cells were grown on melinex polyester film until a confluent monolayer had been formed (5–7 days). The cells were then exposed to rotenone (10 µM) in a HEPES-buffered solution (see 'Endocytosis Assay') for 1 h at 37°C. At the end of the experiment, the drug was washed off and primary fixation was performed in 1% paraformaldehyde/1.5% glutaraldehyde in phosphate-buffered solution (PBS). Following a wash with PBS, secondary fixation was performed using 1% osmium tetroxide/1.5% potassium ferricyanide in PBS. The cells were then thoroughly washed with distilled water, followed by dehydration through graded ethanol (30, 50, 70, 90 and 100%); they were infiltrated with a 50:50 mixture of 100% ethanol/epoxy resin (Lemix, TAAB Laboratories Equipment Ltd., Aldermaston, UK) and finally an overnight infiltration was performed with 100% resin.
Pieces of the melinex were then flat embedded with the cells face up in liquid resin. These were polymerized at 70°C overnight, and the melinex was then stripped away from the cells leaving them embedded in the resin. The resin blocks were trimmed down and a secondary embedding was carried out with the cells face up. Using a Reichert Ultracut E ultramicrotome, semi-thin sections were cut with glass knives and stained with toluidine blue. Ultra-thin sections were cut with a diamond knife picked up on copper grids and stained with uranyl acetate and Reynold's lead citrate. The grids were viewed with a Philips CM120 transmission electron microscope and digital images collected with an AMT digital camera.
The results are presented as means ± SEM. One-way analysis of variance was used to investigate statistical differences among the study groups. Where differences were found, individual groups were compared by an unpaired t test. A p value of <0.05 was taken as significant.
OK cells form a confluent monolayer when grown in normal culture conditions: they endocytose FA when incubated at 37°C, which can be measured as a progressive increase in fluorescence signal over 1 h (fig. 1a). This uptake was, at least in part, mediated by the megalin receptor, as it was inhibited by receptor-associated protein, an established inhibitor of megalin-mediated endocytosis [19] (fig. 1b). To assess the polarity of FA uptake, OK cells were grown on Transwell inserts until confluent. Significantly greater uptake of FA was observed after 1 hour when FA was applied to the apical membrane than when applied to the basolateral membrane (fig. 1c). This is consistent with the situation in the PT in vivo, where LMWP endocytosis is thought to occur mainly at the apical membrane. From here on, endocytosis experiments were performed with OK cells grown as monolayers on coverslips, where the cell membrane exposed to ligands is predominantly apical.
|Fig. 1.a Mechanism and polarity of FITC-albumin uptake in OK cells. OK cells endocytose FITC-albumin progressively over 1 h. b Uptake is partly megalin-mediated, as demonstrated by a reduced fluorescence signal after 1 h in the presence of the megalin receptor inhibitor receptor-associated protein (RAP). c Experiments performed on OK cells grown on transwell inserts revealed that apical uptake was greater than basolateral uptake. Values given are mean fluorescence (± SE) per ×40 objective field (a and b: n = 15, c: n = 10). * p < 0.005.
To ensure that exposure of OK cells to inhibitors of the mitochondrial RC does not result in rapid necrotic cell death within the time period of the endocytosis assay, a standard Hoechst/propidium iodide assay was performed. After 2 h (twice the length of the endocytosis assay period) the proportion of dead cells observed compared with the control (0.17% ± 0.17 per ×40 objective field, n = 5) was not significantly increased following incubation with rotenone (10 µM) (2.39% ± 0.73, n = 5, p > 0.9), cyanide (1 mM) (0.20% ± 0.20, n = 5, p > 0.9) or 2-DG (5 mM) (1.62% ± 0.50, n = 5, p > 0.9) (fig. 2a).
|Fig. 2.a The effect of RC inhibition on cell death and total FITC-albumin uptake in OK cells. Necrotic cell death in OK cells was measured using a Hoechst/propidium iodide assay. There was no significant increase compared with control in the percentage of dead cells after 2 h of incubation at 37°C with rotenone (10 µM), cyanide (1 mM) or 2-DG (5 mM). Values given are mean percentage (±SE) of propidium-iodide-positive cells per ×40 objective field (n = 5). b Uptake of FITC-albumin in OK cells over 1 h at 37°C was inhibited by a small, but significant, amount by the RC inhibitor piericidin (10 µM). No significant decrease was observed with rotenone (10 µM) or cyanide (1 mM). 2-DG (5 mM) caused a much larger impairment of FITC-albumin uptake. Values given are mean fluorescence (± SE) per ×40 objective field [n = 20 for all experiments, except 2-DG (n = 15)]. * p < 0.05, ** p < 0.001.
The control fluorescence signal emitted by OK cells per ×40 objective field (assigned the value of 100%) after incubation for 1 h at 37°C with FA was reduced by a small amount by incubation with the RC inhibitors rotenone (10 µM) (92.8% of control signal ± 2.4, n = 20), piericidin (10 µM) (89.5% ± 2.0, n = 20) or cyanide (1 mM) (93.7% ± 1.8, n = 20) (fig. 2b), although only the results obtained with piericidin achieved statistical significance (p < 0.05). This shows that inhibition of the RC in OK cells has little or no effect on receptor-mediated endocytosis at the apical membrane in OK cells. However, incubation with 2-DG (5 mM) produced a large and significant decrease in fluorescence signal (51.6% of control signal ± 1.7, n = 15, p < 0.001), demonstrating that blockade of glycolysis in this cell model causes a far more potent inhibition of endocytosis than RC inhibition.
To investigate why blockade of glycolysis had a much greater inhibitory effect on FA uptake than RC inhibition, we compared the relative dependence of ATP generation on aerobic versus anaerobic metabolism in OK cells. OK cells were loaded with the fluorescent dye magnesium green; intracellular free magnesium concentration (and so the magnesium green fluorescence signal) changes inversely in response to changes in cytosolic ATP levels. The cells were simultaneously loaded with TMRM, to provide a measure of Δψm, and changes in the cytosolic signal were measured to monitor the redistribution of TMRM from mitochondria to cytosol in response to falling Δψm.
Inhibition of the RC with rotenone (10 µM) did not change the magnesium green signal, suggesting a minimal effect on the cytosolic ATP level (fig. 3a). However, rotenone did cause a rise in the cytosolic TMRM signal (fig. 3a), consistent with depolarisation of Δψm, and redistribution of the dye from the mitochondria to the cytosol. This change provided a positive control and indicated that the drug was having its expected effect (i.e. inhibiting RC activity). In response to subsequent inhibition of glycolysis with IAA (1 mM), the magnesium green signal increased, indicating a fall in cytosolic ATP levels. IAA caused an additional rise in the TMRM signal, consistent with further depolarization of Δψm. This reflects the dependence of Δψm on the 'reverse' activity of the F1FO-ATPase, which is maintained by the hydrolysis of glycolytic ATP when the RC is blocked [26].
|Fig. 3. The effect of RC inhibition on cytosolic ATP levels in OK cells. a Rotenone (10 µM) caused an increase in cytosolic TMRM signal, consistent with inhibition of the RC and depolarisation of Δψm. However, IAA (1 mM) had a much greater effect on magnesium green (Mg green) signal than rotenone, implying that cytosolic ATP is predominantly derived from anaerobic metabolism in OK cells. Values given are mean nuclear fluorescence signal (± SE) from a total of 154 (TMRM) and 126 (Mg green) cells. To corroborate these findings using a different technique for measuring cytosolic ATP levels, OK cells were transfected with luciferase targeted to the cytosol (Cyt-Luc). Cells emitted a luminescence signal following the addition of luciferin (50 µM), the intensity of which is ATP dependent. b A representative trace is depicted, showing that RC inhibition with rotenone (10 µM) produced only a small decrease in signal intensity, while a much larger decrease occurred following blockade of glycolysis with IAA (1 mM). c Mean normalized values (± SE) (1 = maximum signal after luciferin and 0 = zero counts) from 11 experiments are given. * p < 0.01.
To corroborate the data obtained with magnesium green/TMRM, the effect of RC inhibition on cytosolic ATP levels was also investigated using a luciferin-luciferase assay. Addition of luciferin to OK cells transfected with luciferase targeted to the cytosol produced a large rise in luminescence, reflecting high transfection efficiency (fig. 3bi). Addition of rotenone (10 µM) caused only a small decrease in luminescence signal to a normalised luminescence value of 0.735 ± 0.009 (where 1 = maximal signal after luciferin and 0 = zero counts, p < 0.01) (fig. 3bii). Following addition of IAA (1 mM), the normalised luminescence fell to just 0.185 ± 0.003 (n = 11 experiments, p < 0.01), suggesting almost complete depletion of ATP.
Taken together these results suggest that the OK cells are predominantly dependent on anaerobic glycolysis for ATP production and that RC inhibition has little effect on cytosolic ATP levels, which might explain the small effect of RC inhibition on FA uptake in OK cells.
When imaging the uptake of FA in OK cells it was noted that while there was very little difference between the various RC inhibitors in their effect on total uptake, there was a marked difference in the pattern of uptake, particularly for rotenone. Neither piericidin (10 µM), another inhibitor of complex I, nor cyanide (1 mM) changed the control pattern of FA uptake; however, a striking difference was observed in the presence of rotenone (10 µM) (fig. 4a). The effects of rotenone on the pattern of FA uptake were unlikely to be due to an impairment of substrate utilization at complex I because piericidin did not have the same effect. Furthermore, bypassing the inhibition of electron flow at complex I with the addition of the complex II substrate methyl-succinate (5 mM) (which should restore normal RC function and ATP production) did not affect the uptake pattern observed with rotenone.
|Fig. 4.a The effects of rotenone on the pattern of uptake of FITC-albumin/FITC-dextran, ROS production and cell structure. The control pattern of uptake of FITC-albumin in OK cells was markedly altered by exposure to theRC inhibitor rotenone (10 µM) but was not affected by exposure to another complex I inhibitor, piericidin (10 µM), nor by the complex IV inhibitor cyanide (1 mM). The pattern of FITC-albumin uptake observed in the presence of rotenone was not altered by incubation with either the RC complex II substrate methyl-succinate (5 mM) (which will bypass inhibition at complex I) or the anti-oxidant MnTBAP (50 µM). Images shown are representative ×63 objective fields of OK cells incubated with FITC-albumin or FITC-dextran at 37°C for 1 h. b Mitochondrial ROS production in OK cells was measured using the dye mitosox; the rate of change of fluorescence signal was taken to be proportional to the rate of ROS production. Rotenone (10 µM) caused a significant increase in the rate of ROS production compared with the control. However, this increase was obliterated by the anti-oxidant MnTBAP (50 µM). No significant difference from the control rate of production was observed after addition of piericidin (10 µM), while cyanide (1 mM) significantly decreased this rate. Values given are mean rates of change in fluorescence signal per ROI, expressed as a percentage of the control rate (± SE) from 163 cells (rotenone), 225 cells (rotenone + MnTBAP), 218 cells (piericidin) and 140 cells (cyanide). * p < 0.001 compared to control. c An uptake pattern of FITC-albumin similar to that caused by rotenone was observed in OK cells following exposure to colchicine (10 µM), an inhibitor of microtubule assembly. In cells exposed to rotenone (10 µM), the control pattern of FITC-albumin uptake was restored by the presence of taxol (10 µM), a microtubule-stabilising agent. Representative images are depicted of single OK cells (scale bar = 2 µm) and ×63 objective fields (scale bar = 20 µm) following incubation with FITC-albumin at 37°C for 1 h. Rotenone also caused an abnormal pattern of FITC-dextran uptake, demonstrating that the effects applied to fluid phase as well as receptor-mediated endocytosis. d Electron micrographs of OK cells confirm that the cells form a confluent monolayer in culture, with an apical brush border (BB). Following exposure to rotenone (10 µM) for 1 h, extensive structural damage was observed in the cells with loss of the brush border and the appearance of numerous vacuoles (V) and abnormally shaped mitochondria (M).
For fluid-phase endocytosis the control pattern of uptake of FD was similar to FA; the majority of FD appearing in the vicinity of the apical brush border (fig. 4a). Rotenone (10 µM) also affected the pattern of uptake of FD, similar to its effects on FA uptake. This suggests that the striking effects of rotenone on FA uptake are probably due to widespread structural changes within the cell that disrupt the uptake machinery, rather than a specific or direct effect on megalin/cubulin receptor-mediated endocytosis.
We postulated that the effects of rotenone may have been due to excess ROS production, since complex I is a potent site of ROS generation in mitochondria. However, the anti-oxidant Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP – 50 µM) had no effect on the pattern of FA uptake seen with rotenone (fig. 4a).
As a positive control to ensure that 50 µM of MnTBAP was sufficient to prevent any rise in ROS production caused by rotenone, we measured mitochondrial ROS production in OK cells by loading them with the ROS-sensitive dye mitosox, a mitochondrially targeted derivative of hydroethidium. The mitochondrial localisation of the mitosox signal was evident, and the fluorescence signal increased progressively with time; the rate of increase was taken to be proportional to the rate of ROS production. Rotenone (10 µM) caused a significant increase in the rate of change of mitosox signal (rate after rotenone = 282 ± 21% of the control rate, n = 163 cells, p < 0.001) (fig. 4b). This was prevented by 50 µM MnTBAP (rate post rotenone and MnTBAP 103 ± 16% of the control rate, n = 225 cells). Thus, 50 µM of MnTBAP completely prevents the increase in ROS production seen in response to rotenone, but does not alter the effect of rotenone on intracellular FA distribution, suggesting that the effects of rotenone on the pattern of FA uptake in OK cells were not mediated by ROS. The rate of change of mitosox signal was not significantly increased by piericidin (10 µM) (99 ± 4% of the control rate, n = 218 cells, p > 0.9), another RC complex I inhibitor, implying that rotenone-induced ROS production depends on the site of action of this drug at complex I. The rate of change in mitosox signal was significantly decreased by cyanide (1 mM) (57 ± 5% of control rate, n = 140 cells, p < 0.001), indicating that blockade of RC at complex IV lowers the rate of ROS production in OK cells.
Since neither methyl-succinate nor MnTBAP prevented the effects of rotenone on the pattern of FA endocytosis, and the RC inhibitors piericidin and cyanide did not produce the same changes as rotenone, we wondered if the effect of rotenone on endocytosis was extra-mitochondrial. There are published data suggesting that rotenone can depolymerise microtubules [27,28,29], in addition to its effects on the RC. Colchicine is an established inhibitor of microtubule polymerisation. A pattern of FA uptake similar to that observed with rotenone was seen in response to colchicine (10 µM); furthermore, co-incubation with the microtubule-stabilising agent taxol (10 µM) with rotenone (10 µM) produced a pattern of FA uptake similar to control (fig. 4c). These results suggest that the effects of rotenone on the pattern of FA uptake in OK cells are due to inhibition of microtubule polymerisation.
The structural effects of rotenone on OK cells were investigated further by electron microscopy. Control images showed that OK cells form a confluent polarised monolayer in culture with an apical brush border (fig. 4d), which is clearly visible, but not as well developed as the native PT brush border in vivo. Addition of rotenone (10 µM) caused widespread structural damage, including loss of the brush border and the appearance of numerous large vacuoles, and abnormally shaped and swollen mitochondria.
Inhibition of the mitochondrial RC in OK cells produced little or no effect on the total uptake of FA, which is perhaps surprising given the seeming vulnerability of the PT to mitochondrial dysfunction in vivo. Several clinical observations support a role for mitochondrial dysfunction in the pathogenesis of the renal Fanconi syndrome: (1) the typical renal presentation of children with mitochondrial cytopathy is Fanconi syndrome [5,30]; (2) drugs known to be toxic to mitochondria (including anti-retroviral therapy) can cause reversible Fanconi syndrome [31]; (3) Fanconi syndrome has been reported in primary biliary cirrhosis, a condition characterized by the presence of anti-mitochondrial antibodies [32]; (4) the toxic metabolites that accumulate in cystinosis [33] and tyrosinaemia [34] (both established causes of Fanconi syndrome) inhibit complex I of the RC in vitro; (5) blockade of the RC inhibits sodium and phosphate transport in isolated rabbit PT [35], and cystine loading (an in vitro model of cystinosis) leads to ATP depletion and impaired fluid reabsorption [36]. However, unlike the native PT, we have shown that OK cells have a significant capacity to generate ATP anaerobically, which probably explains their resistance to RC inhibition.
Immortalised cells in culture tend to become increasingly anaerobic with time [37], as is true of many cancer cells (the Warburg effect [38]). Accordingly, blockade of glycolysis in OK cells produced a larger decrease in cytosolic ATP levels than RC inhibition and also inhibited FA uptake to a greater extent, suggesting that ATP might play an important role in receptor-mediated endocytosis in the PT. A previous study investigating the effects of metabolic inhibitors on fluid-phase endocytosis in OK cells also reported that IAA caused a bigger drop in ATP levels than cyanide, consistent with our findings [39]. IAA is an alkylating agent that interacts with thiol groups on proteins and, therefore, may have effects on enzyme systems other than glycolysis [40]; nevertheless, this does not detract from the finding that RC blockade had only a relatively small effect on ATP concentration.
Previous studies of both renal and non-renal cell lines indicate that a change in cytosolic ATP level can have a significant effect on receptor-mediated endocytosis. The endocytosis and recycling of transferrin and its associated receptor in the MDCK renal cell line, which is more distal nephron-like in its properties, was found to be dependent on ATP levels [41]; furthermore, uptake of radio-labelled transferrin in both K562 and Hela cells can be inhibited by ATP depletion [42]. In the Cos-7 cell line the chloride channel CIC-2 is upregulated at the cell membrane following ATP depletion, which decreases endocytic retrieval of the channel from the plasma membrane [43]. Similarly, in human astrocytoma cells (1321N1), endocytic downregulation of β-adrenergic receptors in response to catecholamines is ATP-dependent [44].
Several intracellular mechanisms involved in the process of endocytosis are thought to require ATP, such as the exchange of free and bound clathrin in clathrin-coated pits [45]. An intact actin cytoskeleton is important for endocytosis [46], and actin polymerisation is ATP dependent (ATP bound to monomeric [G] actin undergoes hydrolysis when polymeric [F] actin is formed [47]). Trafficking of endocytotic vesicles along microtubules by the motor proteins kinesin and dynein also requires ATP [48]. In addition, for receptor recycling, dissociation of receptor and ligand inside endosomes must occur, a process that requires acidification via an ATP-dependent proton pump and chloride/proton antiporter [49,50,51]. Disruption of this acidification process leads to impaired protein endocytosis [52].
To get around the problem of immortalised cell lines becoming anaerobic, strategies have been adopted to encourage a switch to aerobic metabolism. The methods tried have included shaking culture flasks to increase aeration and lowering culture medium glucose concentration to reduce the drive toward glycolysis [53]. However, it is unclear how successful these approaches are. An alternative manoeuvre is to use primary cell cultures, although freshly isolated PT cells tend to lose their phenotype within a few passages. Direct visualisation of LMWP endocytosis in the PT is now possible in vivo using multi-photon microscopy [54], but this is a technically demanding approach.
To investigate other potential mediators of the effects of RC inhibition on endocytosis (apart from changes in ATP levels), we measured the effects of RC inhibitors on mitochondrial ROS production in OK cells. Inhibiting the RC with piericidin did not lead to a significant increase in ROS production, while cyanide decreased ROS production. An earlier investigation of the effects of cyanide on ROS production in another renal cell line, LLC-MK2, reported an increase in ROS production and oxidative stress [55]. However, cyanide exposure was much longer (4 h) and the dye used to measure ROS (dichlorofluorescein) is not specific to mitochondrial ROS production. This paper also reported a significant decrease in cytosolic ATP levels in response to cyanide, suggesting a much greater dependence of LLC-MK2 cells on aerobic metabolism compared with OK cells. From a metabolic standpoint, LLC-MK2 cells seem a more physiological model of PT cells in vivo. However, they exhibit a limited capacity for albumin endocytosis when compared with OK cells.
In contrast to piericidin, inhibition of complex I with rotenone produced a large increase in ROS production. Differences between rotenone and piericidin in ROS generation at RC complex I have been reported before [56], although both are thought to share a common binding domain with overlapping sites [57], differences in the effects of the two reagents may result from different conformational changes induced in the complex on binding either reagent [58]. Much still remains to be done to elucidate the behaviour of complex I and the mechanisms of ROS generation at this site, particularly in different tissues.
Rotenone had marked structural effects on OK cells, causing widespread cellular damage and leading to an altered pattern of both FA and FD uptake. Rotenone is a much used reagent, both as an experimental inhibitor of complex I of the RC and as a commercial pesticide. It is thought to block the electron flow at the ubiquinone-binding site of complex I, leading to an increased NADH/NAD+ ratio at the 'upstream' flavin-binding site, which favours increased ROS production [59]. Another (and less well-known extra-mitochondrial) effect of rotenone is inhibition of microtubule polymerization [27]. The similar effects of rotenone and colchicine on FA uptake in OK cells suggests that rotenone's toxicity is due to microtubule inhibition. This is also consistent with the protective effect of the microtubule-stabilising agent taxol. Microtubules are part of the cell cytoskeleton and are important in trafficking ion transport proteins in polarised cells to the appropriate cell membrane [60]. Injection of colchicine in rats causes significant structural abnormalities in the PT on electron microscopy [61], and inhibition of microtubule polymerisation has been reported previously to reduce albumin uptake in OK cells [46].
In summary, in spite of their widespread use as an in vitro cell model of LMWP uptake in the PT, OK cells are very different metabolically from native PT in vivo; although they show avid and consistent endocytosis of albumin, the underlying mechanisms might not accurately model those occurring in vivo. This may be important, because our results suggest that FA uptake in OK cells is ATP-dependent. Moreover, rotenone should be used with caution in cell culture studies of mitochondrial function, since it causes a much greater increase in mitochondrial superoxide production than either piericidin or cyanide, and it has significant extra-mitochondrial effects on microtubule polymerisation.
The authors are grateful for assistance from the Division of Medicine, UCL. This work was supported by grants from Kidney Research UK and the Wellcome Trust. M.C. was supported by a Marie Curie-Intra-European Fellowship.
Dr. A.M. Hall
Centre for Nephrology, UCL, Royal Free Campus
London NW3 2PF (UK)
Tel. +44 207 679 7127, Fax +44 207 813 0530
E-Mail firstname.lastname@example.com
Received: November 24, 2009
Accepted: February 24, 2010
Published online: May 13, 2010
Number of Print Pages : 11
Number of Figures : 4, Number of Tables : 0, Number of References : 61
Nephron Physiology
Vol. 115, No. 2, Year 2010 (Cover Date: June 2010)
Journal Editor: Kleta R. (London)
ISSN: 1660-2137 (Print), eISSN: 1660-2137 (Online)
For additional information: http://www.karger.com/NEP


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Soria Announces the Addition of Lipoatrophy Repairing Treatment to the Common Services Portfolio of the National Health System...France. August 4, 2008 -- Currently, different surgical techniques have proven to be effective in the treatment of facial lipoatrophy (the loss of facial fat), one of the metabolic side effects that increases the sense of stigma among... Read more »
Can Glitazones Reverse Lipoatrophy?...study found that an anti-diabetes drug, called rosiglitazone (Avandia), is able to reverse the loss of fat under the skin ( lipoatrophy ) that some people with HIV experience. While these results are encouraging, they are just the latest in a... Read more »
Transcript of The Body's Live Chat on Lipoatrophy (Fat Loss) With Dr. Joel Gallant... Following is the transcript of The Body's live chat on lipoatrophy (fat loss), which took place on Nov. 1, 2005, at 9 p.m. Eastern Standard Time. The chat was sponsored by Gilead Sciences, moderated Read more »
Lipoatrophy and Antiretroviral Drug Changes...Lipoatrophy is abnormal fat loss, often seen especially in the face, arms, and legs. Sometimes fat inside the abdomen (a different kind of Read more »
Connect With Others
Lipoatrophy but good labs--I really need some help
Posted by StandinOnFaith, 4 Replies...torso and belly bigger. Of course, at the time I was still in denial about my HIV symptoms, so I never saw a doctor. Today, my lipoatrophy symptoms have gotten worse and more frequent. It's not very noticeable to others, except when family... Read more »
Lipoatrophy correction
Posted by ConFe, 1 Reply...I've been told about BioAlcamid to correct lipoatrophy, Does anybody have comments about this product. Also I'd like to know the Doctor's name who is in Tijuana Mexico? a friend of Read more »
Lipoatrophy Center
Posted by Bear60, 3 Replies...here it is. It's something new at the BODY and includes personal interviews. Mostly has to do with reversing the effects of Lipoatrophy with cosmetic surgery I believe: http://www.thebody.com/lipo/index.html Anyone ever had any... Read more »
Posted by Anonymous, 3 Repliescould anyone please tell me how they are dealing with lipo being a woman. I have lost fat in face, leg, butt, arms. I look like a preg. toothpick....my stomach is so big. I have been off HIV medication going on two years now and I am scared to go... Read more »
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Shipping Weight 0.60lbs.
When Leatherman made the Charge TTi, they combined all of the most requested features into one functional tool. The TTi's premium comfort-sculpted titanium handle scales and an S30V® stainless steel clip-point knife to really take this multi-tool to the next level. Who says a multi-tool can't be sexy?
I found this on the Baileys Online website, http://www.baileysonline.com. I thought you might be interested in seeing it for yourself. Please click on the following link to view the page:


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Deadline to Apply: February 15, 2013
The premiere opportunity to introduce talent and new music at IBMA's annual World of Bluegrass conference comes in the form of "official" showcase performances which help the music industry assembled discover emerging new bands, as well as established bands with new music or new personnel.
Are you curious about World of Bluegrass in Raleigh, North Carolina? Want to get a peek at the facilities before arriving in September? Or are you maybe just looking for an excuse just to get away for a weekend? Well here's your chance!
You're invited to join the IBMA Board of Directors, IBMA staff, and all our Raleigh hosts on April 13, 2013 for some hospitality, North Carolina style.


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Re:Start looks safe for another year, with plans afoot to shift the colourful container mall just down the road.
It has been three years since since a massive earthquake hit Christchurch on February 22, 2011.
Next year is set to be the biggest yet for the $650 million-plus redevelopment of Christchurch's earthquake-hit hospitals.
The largest concrete pour in the Christchurch CBD since the earthquakes gets under way at 2.30am on Saturday at Antony Gough's $140 million The Terrace.
The New Zealand Red Cross 2011 Earthquake Commission has allocated $2.3 million to increase youth work and social work services in Canterbury schools until 2016.
The third anniversary of the 22 February Canterbury earthquake marks a change in focus for the New Zealand Red Cross Recovery Programme, from the distribution of grants to the development of sustainable programmes.
Insurance companies in the Christchurch market have been rated among the world's slowest to respond to disaster.
The government gave the Christchurch City Council just three days to negotiate the cost sharing agreement to rebuild the city worth almost $3 billion, says Canterbury Earthquake Recovery spokesperson Ruth Dyson.
CERA has recently discovered evidence of vandals cutting the wire fence on Summit Road and breaching the safety cordon.
Chris Lynch's debut television show 'Lynched' will kick off CTV's autumn schedule. Throughout autumn, new locally-made programmes, new talent and new graphics will update the look of Canterbury's regional broadcaster.
The reopening of the restored Isaac Theatre Royal in Christchurch city centre has been pushed back about three months to October.
A number of events will be occurring in and around the Botanic Gardens and North Hagley Park this weekend, including the Civic Memorial Service on Saturday. As large crowds are expected to attend the variety of events on offer, we …
Have you ever wondered why sometimes traffic jams occur for no obvious reason?
Burwood/Pegasus Community Board has confirmed the members of a Community Advisory Group to aid the completion of a master plan for the recovery of New Brighton.
Planning is almost complete by Christchurch City Council for a Civic Memorial Service to commemorate the third anniversary of the 22 February 2011 Canterbury earthquake. The service will be on the Archery Lawn at the Botanic Gardens on at noon and will include one minute's silence at 12.51pm.
Sydenham Heritage Trust Chairperson, Dorothy Haywood talks to Newstalk ZB's Chris Lynch about the site of the rubble on the corner of Brougham and Colombo Streets.
THIS WORK HAS BEEN POSTPONED – A NEW DATE WILL BE ADVISED ONCE KNOWN Hagley Avenue between Riccarton Avenue and St Asaph Street will be closed to northbound traffic from 7pm – 7am on Wednesday 19th February. A detour will …
Next OVTRK Open Forum:
Release: Writers Centre
Hagley Avenue between Riccarton Avenue and St Asaph Street will be closed to northbound traffic from 7pm – 7am on Wednesday 19th February. A detour will be in place from Hagley Avenue via St Asaph Street to Antigua Street to …
Gerry Brownlee | Building and Construction
Release: Statistics New Zealand
The majority of people who left Christchurch following the city's earthquakes stayed in the Christchurch area.
Organisor of the Older Generations Forum, John Patterson talks to Newstalk ZB's Chris Lynch about how the Insurance companies are taking care of the Elderly.
Christchurch City Mayor Lianne Dalziel talks to Newstalk ZB's Chris Lynch about her State of the City speech she has given.
Gerry Brownlee | Canterbury Earthquake Recovery
The earthquakes of 2010 and 2011 have changed the face and shape of Greater Christchurch – the area that covers the city, and Waimakariri and Selwyn districts.
A blogger for website EQC Truths has been fined $5000 for deliberately disobeying a court order.
A Christchurch drug and alcohol counsellor walking 288 kilometres from Seddon to Christchurch to help raise money for earthquake victims is more than half-way through her journey.
Some of New Zealand's churches will be the focus of an international study intended to help earthquake-proof buildings.
Christchurch Mayor Lianne Dalziel is pushing for urgent action to address the parking problems in the central city.
A substantial section of land offering rural and residential opportunities, strategically located on the expanding south-western border of Christchurch City, has been placed on the market for sale.
The Transport Agency will be undertaking essential maintenance works in the Lyttelton tunnel between the hours of 8pm – 6am on Wednesday 19 February.
Producer for Aftermath: The Triumph of the City, Virginia Wright talks to Newstalk ZB's Chris Lynch about how the city has progressed.
On the third anniversary of the February 22 earthquake, Cantabrians will once again have the opportunity to share their experiences and hopes for the future as part of the River of Flowers commemoration.
I have decided that if all the political parties can start the year with a State of the Nation address, then I should begin the year with a State of the City address.
UC's earthquake archive digitally preserves Christchurch's post-quake Pledge books
Durham Street has two areas of road works that will contribute to delays this week. Just south of the Kilmore Street intersection Durham Street is reduced for a short distance to one lane. In additiona Durham Street is reduced to …
Signatures pledged by more than 25,000 Cantabrians after the February 2011 earthquake have been preserved in a digital archive as a reminder of the city's early reactions to the devastation.
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Reserve your place at Reed Arena for the 2014-15 season. 12th Man Foundation priority seating includes the best seats in Reed Arena and provides a charitable contribution to support scholarships for Texas A&M student athletes.
We're here to help you get in the game, click on the seating chart link to the left to view ticket opportunities. Should you have any questions, please call 1-888-99AGGIE.
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Frankfort, KY – The federal government has awarded Kentucky's agriculture department nearly $240,000 in grants to boost the competitiveness of specialty crops in the state. Agriculture Commissioner Richie Farmer says the grants will help producers expand and market their specialty crops. The agriculture department will have nearly $62,000 to hire a marketing specialist for two years to focus on developing specialty crops. Another $80,000 will go toward research on controlling pests on organic specialty crops. The money will also go toward educating farmers on growing blackberries, currents, edamame and other crops.


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beauty blog | makeup reviews | cosmetic-candy.com
I had to share this beautiful picture with you from the Guardian website.
This little elephant has the most amazing lashes I have ever seen, although I am sure they aren't as soft as human lashes!
My two butterballs have rather lovely soft squidgy skin - not an easy thing as small children are … [Read More...]
Having been a long time RMK fan (Creamy Foundation FOR EVER), I am always happy to see their new … [Read More...]
I am all for a new lash mascara/primer/base. I have tried many many many and more often than not I … [Read More...]


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AHL announces suspensions
SPRINGFIELD, Mass. … The American Hockey League today announced that Grand Rapids Griffins left wing Trevor Parkes has been suspended for one (1) game as a consequence of a boarding incident in a game at Rockford on Jan. 11.
Parkes will miss Grand Rapids' game Wednesday (Jan. 16) at Charlotte.
* * *
The American Hockey League today announced that Syracuse Crunch right wing Richard Panik has been suspended for three (3) games as a consequence of an interference incident in a game vs. Rochester on Jan. 12.
Panik will miss Syracuse's games Friday (Jan. 18) at Rochester, Saturday (Jan. 19) vs. Lake Erie and Jan. 25 vs. Binghamton.
- Consistency is key for Chaput and the Falcons
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- Chiarot's dream being realized in Winnipeg
- Playoff Primer: March 26
- Flames' Poirier shining in rookie season
- Crunch to honor NBA's Nationals
- AHL announces suspension
- Sharks' Lerg named Player of the Week


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News archive page
Betty White's Biggest Regret Is So Sweet, It Hurts
At 93 years old, Hollywood icon Betty White has had a lifetime's worth of experiences. She has worked alongside comedic heavyweights like Mary Tyler Moore, become one of the most nominated women in Emmy Award history, and appeared in everything from films to sitcoms to "Saturday Night Live." In her…Huffington Post
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13 Childhood Photos Of History's Most Evil Humans
Little did these photographers know that they were capturing some of the most infamous men of all time.
Jenelle Evans Says She Weighs 108 Pounds: What Are the Teen Mom 2 Star's Weight Loss Secrets?
Teen Mom 2's Jenelle Evans revealed she lost 7 pounds in March, and now weighs just 108 pounds -- details!Us Weekly55 mins ago
Meet Jimmy Page's Very Young New Lady, Scarlett Sabet
Age is only but a number... or is it? Jimmy Page, 71, has been photographed several times now holding hands with 25-year-old actress Scarlett Sabet. For those of you without an abacus, that's a whopping 46-year age difference.Yahoo Celebrity
This Is Why Sarah Jessica Parker Looked So Disgusted With Tom Hanks at the Rangers Game
American Treasure Tom Hanks recently popped up on the Jumbotron at a Rangers game at Madison Square Garden, seated directly in front of Carrie Bradshaw herself, Sarah Jessica Parker. First thought: Why have these two never been in a movie together? Wouldn't that be perfect? NOT IF THIS VINE HAS…ETonline
Celebrities Join the Selfie Stick Craze (13 photos)
Social media stars, including Sean "Diddy" Combs, Kaley Cuoco-Sweeting, and Jennifer Lopez, know how to snap the best selfie. Instead of awkwardly hold their phones away from them while quickly pressing the button, they use a selfie stick to make sure they're always ready for their close-ups.Yahoo Celebrity
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19 Hidden Easter Eggs In Disney Movies
We all love to watch Disney movies over and over again, but did you ever notice those tiny hints in some of the scenes?
D.A. Points breaks PGA Tour's ShotTracker with Texas Open implosion
There's carnage everywhere at the Texas Open but Points' 8-over through three-holes start is something special. The above image is some sort of sadistic artwork, and also ShotTracker's best attempt at capturing the misadventures of D.A. Points on the first green of the Valero Texas Open. This TPC…SB Nation
Banished From 'The Breakfast Club': Actress Recalls the Burn of Getting Cut From John Hughes' Film
Original 'Club' goer Karen Leigh Hopkins on the role that got away.Yahoo Movies
January Jones: My Confidence Disappeared When I Got 'Pretty'
Looks aren't everything. Mad Men star January Jones is obviously a beautiful woman, but in a new interview with Violet Grey, she surprisingly reveals her confidence took a turn for the worse when she started growing into her looks. The 37-year-old actress -- who goes topless in her latest…ETonline
Ben Stiller: Joan Rivers and I Never Made Amends Before Her Death, "She Didn't Like Me"
Ben Stiller spoke with Howard Stern about his feud with Joan Rivers, saying it was "sad" they didn't make up before her death in 2014Us Weekly
- AdChoicesAnswers HollywoodSponsored
Where Are They Now: The Cast of 'My Three Sons''
The 1960's series 'My Three Sons' was a widely popular show featuring the beloved Fred MacMurray as the Douglas family patriarch. Millions of. . .
Harry Styles Crying Over Zayn Malik On Stage Is the Saddest Thing You'll See All Day
On Wednesday, One Direction performed its first show since Zayn Malik revealed he was leaving the band.TV Guide
Jason Aldean And Brittany Kerr Share First Wedding Photo
Jason Aldean and Brittany Kerr's wedding was certainly picture-perfect. Aldean and Kerr married in a beach ceremony surrounded by family and friends Saturday in Playa del Carmen, Mexico. The couple's first official wedding photo appears on the cover of the latest issue of Us Weekly magazine, which…Huffington Post
Ohio mom of decapitated baby faces aggravated murder charge
CINCINNATI (AP) — A woman accused of decapitating her 3-month-old daughter with a chef's knife was indicted Wednesday on a charge of aggravated murder.Associated Press
Fox mismanages the first reveal of The Thing from 'Fantastic Four' reboot
One of the things comic book fans are most interested about this summer is the appearance of The Thing (Jamie Bell) in "Fantastic Four," and now there's an image online from a terrible source, and it raises the question of how Fox would let that happen.Hitfix
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16 Hilarious Celeb Derp Shots
Cross your eyes and check out these flawless shots of your favorite celebs! Then, uncross them for a whole heap of derps in these herpy celeb pics.
Bill O'Reilly's Visit With David Letterman Draws Fire
Bill O'Reilly's victory lap on late-night TV last night was horseradish that CBS late-night host David Letterman should not have served cold to his audience, says one of the media outlets that's been probing claims the Fox News star has made about his early days in journalism. Making his first…Deadline
Zayn Malik Leaves 'One Direction'
After backing out of the "One Direction" world tour last week, Zayn Malik has announced that he is leaving the boy band for good.ABC News Videos
6 Gym Machines That Are Actually Worth Your Time
Nearly every gym has two distinct sections—free weights and machines—and they attract different crowds. Serious lifters do their thing with free weights, and weaklings and wannabes stick with the machines.STACK (non-hosted content)
Amal Clooney Shows Off Her Abs in a Feather Crop Top and Funky Bell Bottoms
Amal Clooney clearly isn't afraid to take risks when it comes to fashion. The 37-year-old human rights lawyer was definitely out to turn heads Tuesday night in New York City, when she went out to dinner with friends in a black crop top adorned with feathers. And if her daring top wasn't enough to…ETonline
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13 Celebrity Wedding Dress Fails
These poor ladies tried a little too hard when it came to their big days, and you will see what we mean.
Billy Ray Cyrus Defends Miley Cyrus' Boyfriend Patrick Schwarzenegger After Wild Spring Break Photos
Billy Ray Cyrus defended his daughter Miley Cyrus' boyfriend Patrick Schwarzenegger after his wild spring break photos made headlines last weekUs Weekly
Enrique Iglesias and Anna Kournikova: Still Afloat After All These Years
He will be her hero, baby (on a boat).Yahoo Celebrity
Josh Groban Responds to Gay Rumors: 'Whatever, Fine'
Despite ongoing speculation, Josh Groban isn't gay -- and not even Ryan Gosling can change that. "If I was gonna be gay, that would've been the moment. That was probably my greatest test, and I failed," the 34-year-old singer (and sometimes) actor says of co-starring opposite Gosling in 2011's…ETonline


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Has anyone tried submitting to this magazine. Granted you may not get rich with what they pay, but it sounds like they are looking for freelance writers.
Here is a link that might be useful: Green Prints
Yes, I submitted an article many years back and Pat liked it and paid me for it then never used it and returned it. Later I entered their contest for Bad Garden Writing and won. Go ahead, laugh. And just TODAY I sent in an article written from the plant's point of view. I don't know if it will fit their editorial format but I was told it may be mid to late February before I get a response.
This is a great little magazine. I have had one article picked up by them and currently have recently submitted another one. The whole process is quite simple. Just follow their online instructions and send in your submission by snail mail. You are right, you won't get rich here but then I don't think Pat Stone is getting rich on his little labor of love either.
I heartily endorse this little magazine. If I have one criticism, it is sometimes a little too warm and fuzzy for me. However, most of its readers would probably say that is the magazine's greatest asset -- and it probably is. It certainly fills a void addressed by no other publication that I know of.


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Is President Bush Behind the Push to Indict Bonds?
Another story I just posted at FanHouse, based on a report from the Mercury News. They're both a must read. In short, Bonds' lawyer, Michael Rains, thinks that President Bush is driving the push for indictment. Funny thing is, I think he might be right. Let's see why: President Bush is a baseball fan. He's…Read More


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It's Met Gala night! Fashion's biggest red carpet of the year! As the stars roll up in their "punk"-themed best (or not), we'll provide you with up-to-the-minute coverage, complete with designer credits and commentary. Click through our slideshow to see Anna Wintour's daisy-print Chanel, Rooney Mara in Givenchy, Kim Kardashian's Met debut, and maybe even a mohawk or two. Enjoy!BEGIN SLIDESHOW
Alexander McQueen
See it »


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Moutain View Toros vs Desert Ridge Jaguars (Full Game: 10/30/14)
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Shilpa Shetty with Rahul Mahajan on the sets of the reality show Bigg Boss after the latter's elimination from the show. (BCCL Photo)
The sixth season of 'Bigg Boss' is full of surprises, the latest being the midnight eviction of actress Delnaaz Irani.
PAMELA ANDERSON SEASON 4 Oomph, appeal and spice — former Baywatch babe Pam exuded it all. Her entry into the house in a translucent white nine yard wonder made fans of the reality show take note of her glam antics. Pamela Anderson, not only impressed her fans in India, but also added some more eyeballs to the show with her glam presence.
After the stupendous success last year, 'Bigg Boss' is once again showing it's charm to the TV buffs with dazzling Shilpa Shetty as it's host. A big comeback with politicians, TV and movie stars, item girls and newsmakers like Monika Bedi, Rahul Mahajan and Jade Goody, the show will surely make for an exciting viewing. The Bigg Boss fans are sure to feel the heat on the show with the very hot and sizzling, Payal Rohtagi. (Photo/Mr.Nalin Solanki)
Here's the fifteen participants who will be locked up for the sixth time to squabble, plot and survive for three months inside the Bigg Boss house. The final participant will be a non-celebrity. Former cricketer and TV personality, Navjot Singh Sidhu enters the Bigg Boss house as a participant for the upcoming season.
Rani Mukherjee, Salman Khan and Vidya Balan in a still from the TV show 'Bigg Boss 4'. (Pic: Viral Bhayani)
While the final list of 'Bigg Boss 4' inmates is still being kept under wraps, this one's already started making waves! Rumours are rife that one of Shah Rukh Khan's look-alike is set to appear as one of the contestants on 'Bigg Boss 4'. Wonder how would the King Khan react to this!
All the seasons of the reality show Bigg Boss have been the talk of the town, thanks to its controversial contestants. Even the producers have managed to rope in some of the most sought after celebs to bring in more and more TRPs to the show. So, here's a list of the celebs, who have been the most talked housemates on the show Sunny Leone: Indo-Canadian pornographic actress Sunny Leone was seen in Bigg Boss season 5. The stunning beauty, whose reality was unknown to the housemates, charmed her way into the hearts of the audience with her seductive pole dance in one of the episodes of the reality show. During her stay in Bigg Boss, filmmaker Mahesh Bhatt approached Sunny and offered her the lead role in Jism 2. Sunny did an item number in Shootout at Wadala and will soon be seen in Ragini MMS 2.
They are the ones who keep the TRPs soaring for popular reality show Bigg Boss. Here's a slide on all love stories that blossomed inside the house: Diandra Soares-Gautam Gulati: Just when it appeared that Diandra and Gautam were seeing each other, some emotional outbursts and strong stands fell their relationship apart. While Gautam had soft corner for the model-actress, Diandra took her own time to seal the relation. Their infamous escapades in the washroom raised many eyebrows.
Salman Khan and Katrina Kaif on the sets of the TV show 'Bigg Boss 4'. (Pic: Viral Bhayani)
Monica Bedi was a contestant in 'Bigg Boss 2'.
Colors announced that Bigg Boss 7 will be starting from September 15, 2013. Here's the list of contestants for this seasons.
Vindu Dara Singh won the 3 season of reality show "Bigg Boss 3" and walked away with prize money of Rs 1 crore. (Pic:Nandu Dhurandhar)
Jaipur-based music composer and singer Sharib Sabri, who has sung some popular numbers in Bollywood, has now been offered season 6 of the baap of all reality shows "Bigg Boss".
Shamita Shetty on the sets of 'Bigg Boss 3'.
Shamita Shetty walked out of reality show "Bigg Boss 3" to be with her elder sister Shilpa who is tying the knot with millionaire beau Raj Kundra on Nov 22.
Akshay Kumar (r) presents a trophy to Ashutosh, the winner of 'Bigg Boss 2', during the grand finale in Mumbai, November 22, 2008. (Reuters/Manav Manglani)
Sherlyn Chopra is evicted from the 'Big Boss 3' and sad she cannot provide the much needed oomph to the house. But the outrageous babe is not out of the house alone, some secrets too came tumbling out along with her. Let's hear the spicy details from her regarding the inmates of the house. (Pic: Maxim)
Salman Khan during the launch of Bigg Boss 8, in Mumbai.
Bollywood megastar Salman Khan interacts with the media during the official launch of India's most popular reality show Bigg Boss 7, in Mumbai. (Pic: Viral Bhayani)
Salman Khan and Anil Kapoor shake legs with Sri Lakan beauty Jacqueline Fernandez, on the sets of TV show 'Bigg Boss 6', during the promotion of the movie 'Race 2'.
Bollywood megastar Salman Khan interacts with the media during the official launch of India's most popular reality show Bigg Boss 7, in Mumbai. (Pic: Viral Bhayani)
The grand finale of 'Bigg Boss 6' that was running for the past few months on Colors channel finally happened and Urvashi Dholakia was announced as the winner. (Pic: Viral Bhayani)
Here's a look at some of the probable contestants of 'Bigg Boss - Season 5': If rumours are to be believed, sultry beauty Shweta Salve has also been approached for the popular TV show.(Pic: Maxim)
'Bigg Boss 4' may soon get into the Guinness Book of World Records for the most number of cases slammed at a TV show. A fresh case has been filed against the content of the programme at the Aligarh district court in Uttar Pradesh.
Riteish Deshmukh and Nitesh Rane of Maharashtra Kalanidhi are set to produce the Marathi version of the reality show 'Bigg Boss'.
Salman Khan during the launch of TV show 'Bigg Boss- Season 6' in Mumbai on September 16, 2012.
Bollywood superstars Sanjay Dutt (L) and Salman Khan (R)perform during an event for the launch of 'Bigg Boss - Season 5' in Mumbai on September 29, 2011. 'Bigg Boss' is an Indian version of reality television show 'Big Brother.' It features celebrity housemates who live together for weeks isolated from outsiders, while vying to avoid eviction and win a cash prize.
'Bigg Boss 6' winner Urvashi Dholakia is all smiles as she flaunts her trophy. (Pic: Viral Bhayani)
Sanjay Dutt at airport as he welcomes sumo wrestler Yamamotayama as the latter to enter 'Bigg Boss' house, in Mumbai. (Pic: Viral Bhayani)
American socialite, television reality star, model and actress Kim Kardashian is being wooed to come on the season 6 of the baap of all the reality shows... obviously for the oomph factor.
Amitabh Bachchan at the press meet of TV show 'Bigg Boss' season 3 at Taj Lands End, Bandra.
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|ESPN.com: AFC South
|[Print without images]
|Gary Kubiak should be counted among the brightest offensive minds in the league.
|Gary Kubiak and Matt Schaub have developed a close relationship after six seasons together.
|"I just think his offensive mind is up there with the best of them," Arian Foster said of his coach.