text:
Noni Juice Health Benefits
Noni Juice – Morinda Citrifolia
You've probably seen noni juice at your local health food store. That has got you to wondering. What are the health benefits of noni juice? That's what we're going to talk about here.
Soon you'll discover what people from all over the world have known. This juice is good for your health. It's something that you should regularly add to your diet.
Let's be clear about something. Noni juice isn't a cure all for everything that ails you. It will however be an important tool in maintaining your health. Which is the most vital thing to your existence.
Known for its scientific name, Morinda citrifolia, noni has a green, almost transparent, rough bark and somewhat unattractive smell when mature.
It is used more than 2000 years as a natural tonic and cure for the disturbed balance of the digestive, intestinal and respiratory system and for immunity.
Lots of studies has documented that plants have many benefits to the body. Thanks to the modern research, we now know more about the abilities of Noni juice.
Noni – the elixir of health
Noni is an adaptogenic plant with volcanic soil. The fruit of this amazing plant contains 18 kinds of amino acids, minerals and many vitamins and enzymes and active substances like proxeronine, serotonin, scopoletin and damnacanthal.
The substance for which the noni juice became known and that is in some ways the foundation of its therapeutic capacity is large molecule proxeronine.
With the help of proxeronine in the intestines creates xeronine, which is necessary for effective human body cell regeneration.
Thanks to its adaptogenic action proxeronine is able to normalize the cells that are functioning improperly. Xeronine and proxeronine show antidepressant activity as enhance the secretion of endorphins, which results in enhancing good mood and euphoria.
Xeronine has triggering mechanism of protection against inflammation in the body and ability to increase the resistance to bacteria and viruses.
Using xeronine, cells can be faster and more successfully renewed, but for each renewal takes time and for this purpose Noni should be used for at least three months.
Noni juice is recommended to all who suffer often from viral infections. It also helps the secretion of digestive juices and reduces a significant part of the problem of the digestive tract. Noni can be useful aid in the treatment of various addictions (smoking, alcoholism, drug addiction…), because noni helps in reducing withdrawal symptoms.
Scopoletin enhances the production of serotonin and melatonin in the body and thus has a positive effect on the condition of the nervous system, preventing mood swings and regulating the sleep cycle.
Serotonin is involved in the solving many diseases such as migraine headache, depression, Alzheimer's disease, etc.
Regular use of noni juice provide their normal levels in the brain, which causes a better mood, better sleep, combating depression and migraine attacks, decreases / regulates hunger which is often a result of stress.
Given that scopoletin in general can be described as a substance for relaxation, the other health benefit is in regulating blood pressure, but also helps with cramps, muscle and menstrual pain, asthma and lung problems etc.
Damnacanthal controls the process of cell division and uncontrolled cell proliferation, as well as the active agent may help to protect against the negative effects of radiation on the human body.
Since taking part in achieving optimal levels of so-called T- and B-cells, enhances the functionality of the immune system. Anthraquinone in noni has antibacterial activity.
Active substances in noni enhance the production of nitric oxide (NO) which is present in almost all body functions, and is necessary for their carrying out.
Nitric oxide improves bowel function, inhibits adhesion of blood platelets (thrombocytopenia), helps the secretion of insulin, growth hormone, but it is also natural remedy for potency.
This simple but effective molecule has the ability to reduce angina pain, prevents the formation of blood clots, and reduces high blood pressure resulting from tension and stress.
Its antioxidant action prevents oxidation of LDL cholesterol, stimulates circulation and blood flow to the tissues, including the reproductive organs and thus affects the quality of your sex life.
Less known is the substance limonene. Limonene substance has strong anti-tumor activity, and as an ingredient in noni juice acts synergistically with proxeronine and damnacanthal.
Two other substance with anti-inflammatory activity are eugenol and asperuloside.
Did you know that noni juice can actually lower your cholesterol?
It can. It's just one of the many reasons why you should be drinking this exotic juice. A lot of you have high cholesterol.
Have you read the side effects of the medication that you're taking? Most of you would rather not take that medication.
For some people controlling their diet and exercising can lower their cholesterol. You should ask your doctor if this is possible. Noni juice will help you lower your cholesterol naturally. Without the side effects of harmful drugs.
Many clinical studies confirmed that Noni juice helps with cholesterol.
One of the keys to a healthy cardiovascular system is the maintenance of healthy cholesterol levels. Studies show that drinking 0.9 to 1.2 ounces of noni juice daily helps keep cholesterol within normal levels.
Heart health is the key to the general well-being of your body. Noni juice proven to help and support a healthy cardiovascular system and helps maintain a good and healthy heart.
Noni juice is super antioxidant?
Antioxidants are vital to healthy life – they help in the defense against harmful free radicals. With its combination of fruit juices rich in antioxidants, Noni juice is one of the world's greatest antioxidants.
Noni Juice supports the immune system?
Stress, pollutants and other complications of everyday life have a serious effect on our immune system. Research shows that Noni juice really supports your immune system and helps it to function normally and to carry out its task easier.
It contains superior antioxidants that help the body get rid of harmful free radicals, increases energy and provides a greater level of physical endurance.
Do you have problems with high blood sugar?
Type two diabetes is a big problem these days. More and more people around the globe have it. It's because of the alarming rates of obesity.
Noni juice will help you regulate your blood sugar levels. The health benefits of this alone are astronomical.
It can be difficult learning how to live with type two diabetes. Discovering that you have it can rock your world. Most people can control their type two diabetes through diet and exercise. Help maintain your sugar levels by drinking noni juice.
Arthritis suffers have a reason to drink noni juice
That's because noni juice has anti-inflammatory properties. Your joints will thank you for having a drink of this wonderful juice.
You'll even notice improvement in your mental function. When was the last time you drank something that helped your brain?
You'll do it each and every time you have a drink of noni juice. You can even drink it to lower your blood pressure.
Have you been feeling blue?
Reach for a drink of noni juice. Serotonin production is increased by noni juice. It's an all natural way to cure those blues that's been holding you back.
People have even used it to help them kick addictions. You've got to be wondering right about now. Why haven't you heard of this before?
The internet has given us all the chance to explore and learn about other cultures. Part of that exploring is learning what people eat. Not just what they eat. But, why they eat it. The world is becoming a smaller place. The secrets of people all over the world are being exposed.
Noni supports natural defenses
Maintaining a healthy immune system is important for the general well being of the body. Without diversity and the daily amount of nutrients, complex immune system will not always function properly.
We also know that everyday life filled with stress can cause weakening of the immune system.
To stay healthy, it is important not only to eat a varied and balanced diet, but to live healthier, more active, maintain a healthy weight, and so on.
Noni juice is a great addition to your living a healthy diet will help you feel great.
NONI Juice increases energy?
Studies show that noni juice increase energy and it also:
- normalizes cell function
- shows antidepressant activity
- protects from inflammation, bacteria and viruses
- helps with detoxification
- contributes to a reduction in blood pressure and sugar
- anticancer activity (study suggest a possible unidentified substance in unpasteurized noni fruit juice that may have a small degree of anticancer activity)
Researches and references
alu, KA West BJ Jensen J Zhou BN. Morinda citrifolia L. Noni has Cholesterol Lowering Potential. Folk Botanical Wisdom: Towards Global Markets. Chiang Mai, Thailand 2006;
Wang, M. Y., Lutfiyya, M. N., Weidenbacher-Hoper, V., Anderson, G., Su, C. X., and West, B. J. Antioxidant activity of noni juice in heavy smokers. Chem.Cent.J. 2009;3:13. Carr ME, Klotz J, Bergeron M. Coumadin resistance and the vitamin supplement "Noni". Am J Hematol 2004;77:103.
Simonsen, J. L. Note on the constituents of Morinda citrifolia. J Chem Soc 1920;117:561-564
Safety of leaves from Morinda citrifolia L. The EFSA Journal 2008;769:1-17.


text:
Courses Offered
Chariton Matosov
13 Jun 2016
Studying at Melbourne Polytechnic has been a great pleasure to me. It had enabled me to escape from my routine ...
Memory Martina Chamunorwa
08 Jun 2016
Very educational. I'm enjoying all the classes.
Bikendra Gurung
27 May 2016
The academic life at Melbourne Polytechnic is actually quite good and lived up to my expectations. The support you get ...
About Melbourne Polytechnic
Melbourne Polytechnic has been delivering vocational training since 1912 and today offers a diverse range of innovative, hands-on TAFE (VET) and Higher Education (Degree) programs to equip students with knowledge and skills to excel in an evolving and challenging marketplace. Melbourne Polytechnic is a government-funded institution and one of the largest vocational education providers in Victoria Australia.
Melbourne Polytechnic offers over 100 courses ranging from Certificates, Diplomas and Degrees to international students. These courses are designed to help them achieve academic and career goals easily. Courses available to international students are nationally accredited under the Australian Quality Framework (AQF) in accordance with the Education Services for Overseas Students (ESOS) legislative framework. Melbourne Polytechnic is registered on the Commonwealth Register of Institutions and Courses for Overseas Students (CRICOS). All courses are recognised internationally.
Currently, there are students from over 70 different countries all over the world. Melbourne Polytechnic's modern training facilities are recognised for being some of the best in Victoria and its long-standing relationships within related industries means students gain industry-relevant skills to be job-ready. International students who have graduated from Melbourne Polytechnic often report how useful the courses have been to their career and higher learning. Experienced teachers and hands-on courses will allow students to gain an invaluable insight and contacts within the chosen career path.
Some Random Courses
Let's see what you may also consider
Diploma of Digital and Interactive Games
Melbourne PolytechnicAustralia
Duration2 yearsIntakesFebTuition fee (entire course)Local-InternationalAUD 22,000
Diploma of Interior Design and Decoration
Melbourne PolytechnicAustralia
Duration1 yearIntakesFebTuition fee (entire course)Local-InternationalAUD 12,400
Bachelor of Accounting
Melbourne PolytechnicAustralia
Duration3 yearsIntakesFeb JulTuition fee (entire course)Local-InternationalAUD 46,800

subset: commoncrawl


text:
This weeks interview come from Kalli Hiller of PortableProfessionals.com. She blogs about their travels as a family of three with an infant, as well as how to sustain a nomadic lifestyle. Read more of her adventures at the blog, or find her on facebook.
What was your travel style like before you had children?
My husband and I traveled for three years nonstop before having children. We lived for 1-3 months in each country before moving on to the next, usually choosing an apartment in just one city to stay in. Our style was, and is, low-key: we go to the gym, grocery shop, and hang out with friends much as we would if we were back in the States. We lived in 5 continents and 27 countries this way, all funded through internet business.
What ages were your children when you started travelling with them?
Our son was born on the road. We found out we were expecting in Guadeloupe, a French-speaking island in the Caribbean. We had to decide where we wanted to have him. For a variety of reasons, we chose Mexico. He was born at a hospital in Mexico City. Looking back, that was a very scary time with a lot of unknowns. It turned out just fine though. We'd do it again. Our child knows nothing but this lifestyle.
Why did you decide that exposing your children to travel was important?
Because travel is necessary for my sanity, and my child needs a sane mom.
Although I believe there will be many other benefits—from gaining an understanding of how the world works, to growing in compassion and tolerance, to becoming more confident and gaining life skills—right now Ryder is just a baby, and he doesn't need much more than his mom and dad and milk right now. Overall, I hope that it will lead him to being a free thinker.
What is your travel style now that you travel with children? How did it change from before you were a parent?
Ryder's not even a year old yet, so we're still exploring this one. He's been to four countries and six US states so far though, and to be honest our style hasn't changed too much. We like slow travel, we're in no hurry, and we need a place to work with internet since we fund our travels with internet businesses. We like to meet locals and make connections with people no matter where we are, and in some ways that's easier now with a baby because people will come up and talk to you more often. I just get up earlier and go to bed earlier than I used to, but that's unrelated to travel. Just part of the package deal of being a parent!
What has been the most difficult thing about travelling with children?
In New Zealand we lived out of a somewhat non-spacious campervan. Doing the campervan thing has its benefits with a baby: for example, babies love riding in cars and he got to do a lot of this from day to day. At night, though, he woke up easier because we were all in such a small space. Here in Tahiti, we only have a scooter, which you can't take a baby on, so I've ended up having to walk much more than I would have previously. Those things are just part of the experience though—that kind of difficulty is more than made up for how much more fun having a baby along is—so really the hardest part is the fact that the baby is growing up away from family. Skype helps but it still falls short. However, most Americans these days aren't going to be living very close to family anyway. Learning to build long distance relationships is part of most extended families these days.
What has been the most rewarding part of travelling with children?
I like traveling with our baby better than I did traveling as a couple. And I love couple travel! For me it's because travel can be an essentially selfish thing—the sights you see, the journey you take, while very personally rewarding, can be somewhat inward-focused. Having a child means getting to share those experiences with someone else, and watching them grow and be positively influenced by travel is exciting. Ryder's still just a baby, but I think one effect of travel already is that he's very comfortable being held and interacting with strangers.
What is your favourite travel memory of travelling as a family?
So far the best has been to watch people from all different cultural backgrounds interact with our baby. I like watching him study their faces and have him hear different languages and accents; and having him see people of a variety of ethnic backgrounds. The Indian owner of the restaurant in Greymouth, New Zealand snapping and grinning at him; the Mexican ranch owner carrying Ryder around the streets of Miguel de Allende; the French-speaking Tahitian mothers saying "Coucou!" to him and asking to hold him. Ryder just basks in that kind of attention.
What tips would you offer parents when it comes to travel with children?
My biggest tip would be just to do it and do it often. The difficulties are very overstated by people in my experience, and the more and the earlier you do it the easier it will be.
Also, try to choose family-friendly accommodation. If it's too quiet, you might get some complaining neighbors when your baby is adjusting (read: crying uncontrollably) to a new place. Shared bathrooms are really no longer ideal with a baby.
Are there certain destination or holiday types you would recommend for parents travelling with children?
We've really enjoyed New Zealand with a baby. We just drove for hours while looking out the window at stunning vistas as Ryder napped in the seat next to us. There's so much outdoors/adventure stuff to do here, I'm sure we'll be back when our kids are older so they can do the bungy jumping and jet boating with us. This time around, we just handed the baby off to an employee while we did our adventure sports—they were very obliging.
And finally – what are you upcoming travel plans?
We're planning on traveling indefinitely. Although we've been known to change countries on a whim, we are planning Tahiti—New Caledonia—Vanuatu—Australia for the next 6 months. We'll have gotten to know the South Pacific at that point. After that it's hard to say, but one thing is certain: meeting with other traveling families is paramount to us, because we want Ryder to have friends who live like him.


text:
This baby bear was clinging to his dying mom when rescuers arrived.
Poachers do not discriminate when it comes to the animals they kill, which means they often kill mothers and their babies when out hunting. Animals are poached for a number of reasons, but sloth bears are killed because humans are taking more and more of their natural habitat every year and then set up traps so the wild animals don't encroach on the land that was once theirs.
For this baby sloth bear in particular, his mom walked into a high-voltage wire that was set up around a tiger reserve in India, where the mom died from electrocution despite rescuers attempting to save her. In the saddest display of despair and proof of the inhumanity of these traps, the 10-week-old baby sloth bear crawled on top of his mom and refused to leave her side after she died in front of him.
When rescuers found the pair, the baby was nursing from his mom's dead body; this is something that no baby, whether they are human or not, should ever be forced to do.
The only silver lining to the story is that the baby was found in time to save him and that he, somehow, managed to avoid being electrocuted, though he suffered from some burns. He was named Mowgli and taken to Agra Bear Rescue Facility, which is run by the International Animal Rescue and Wildlife SOS.
"It will take a considerable amount of time for this baby bear to fully recover from this painful and traumatic experience, but we are there for him at every step of the way by giving him all the love and care possible, to help him grow into happy and healthy bear," according to Wildlife SOS.
This certainly isn't the first time that sloth bears have been targeted, but it is one of the cruelest methods that rescuers have ever seen. Killing this endangered species is illegal, and sniffer dogs were brought to the scene to find those behind the electric trap. Six men were arrested for the crime and are awaiting trial.
If rescuers had not found the mom and bear cub, it's likely that the men would have taken both back with them and eaten the mom and sold the baby to use for baiting or to become a "dancing' bear." Bear baiting is when a bear cub is used to teach dogs how to be aggressive for dog fights and the baby bears are brutally murdered by the dogs. The entertainment industry for these bears is cruel as well, as they are forced to perform based on fear and then locked up in cages for days when not entertaining.
The practice of poaching is getting worse, as Lis Key, PR and communications manager for IAR, explained to The Dodo. She said,
"Bears and other wildlife are left exposed and vulnerable — and struggle to find food in their dwindling territory. Wild animals then stray into villages and onto farmland in search of food, trampling or eating people's crops." She added, "In retaliation, people set snares for wildlife and trap all kinds of unwitting victims such as pigs, deer and, as in this case, wild sloth bears," she explained. "Whatever the animal, the poachers eat them as bushmeat."
Mowgli the bear is adjusting well, though he is still traumatized and will need to be cared for at the facility for the rest of his life. By killing his mother while he was at such a young age, the rescue groups point out that they took not only his mother but his freedom as well.
"This young bear has been robbed of his chance to life a life of freedom in the wild as it is in the first 18-24 months of their lives that sloth bear cubs pick up the skills and natural instincts from their mothers that they require to survive in the wild. He is scared and yearning for his mother's presence and is currently at our separate cub weaning unit under the full time care of a keeper."
Though he will never again experience life in the wild, he will live a happy life at the facility with other bears just like him that need to be cared for. If you want to make a donation to help care for this bear, you can do so here.
What are your thoughts on this story? Please share, like, and comment on this article!
This article (Poachers Killed This Sloth Bear's Mom, And He Didn't Let Go Of Her Until He Was Rescued) is free and open source. You have permission to republish this article under a Creative Commons license with attribution to the author and True Activist.


text:
LINE-X Truck Bed Liners Review Sep 27, 2016 by Cedric (Driver Weekly)
There are many choices for spray-on bedliners for your pickup, including those that are provided by the manufacturer and aftermarket options. These bedliners are a good way to protect your vehicle's truck bed from damage, preserving its value. If you're spending money on a new bedliner, LINE-X is among the most popular brands and for good reason.
Lifetime Warranty
One of the things that truly set LINE-X bedliners apart from the competition is that they come with a lifetime warranty. The guarantee says that the liner will not flake, bubble, or crack. If it does begin to do one of these things, it will be repaired by the trained technicians. If damage happens to occur that isn't covered by the warranty, it can also be easily repaired for an affordable price.
Application Process
Instead of just letting you spray the liner on yourself, LINE-X bedliners are applied by technicians with a significant amount of training. Instead of trying to complete as many liners as possible, they take their time and pay attention to detail. They will verify the tolerances of each bedliner, so you get the ideal thickness consistently across your truck bed. The coating is typically thin, around a quarter of an inch. This is ideal since it will follow the contours of the truck bed, keeping the factory appearance you are used to. The thin coating also allows the LINE-X bedliners to create a rough surface. When installed by a professional, it will be run-free since it mixes right in the spray gun and sets within three to five minutes.
Rigorous Testing
LINE-X can offer a lifetime warranty on its products because they have undergone extensive testing throughout the years. LINE-X tests everything from strength to durability and does so in creative ways. The company takes suggestions from customers and scientists and has shown that its bedliners hold up to the toughest tests.
Tough Surface
The only thing to keep in mind is that the high durability of the LINE-X bedliners is partially due to their hard texture. This means that it may feel tough on your knees when you are loading cargo. It also means you have to be careful with more sensitive cargo, such as finished wood surfaces. When transporting a delicate cargo, you may want to lay down a softer protective material. However, the harder material does mean that the system holds up very well over time, with little damage. Like all bedliners, the products made by LINE-X will fade after several years due to their exposure to sunlight. You will still have to work hard to protect your liner from UV rays since it tends to fade more quickly than your truck's paint. Because of this, it can be worth it to pay a little more for UV protection in your LINE-X liner. LINE-X does offer several options that include this UV protection, and their custom color abilities mean that your liner can match your vehicle's paint.
Protecting your truck bed can increase the useable life of your vehicle and maintain it's appearance. LINE-X offers one of the best protections in the field, especially for heavier and rougher loads.


text:
Riders (active)
David Graf
Adrian Kiener
Benjamin Kistner
Simon Waldburger
Adrian Weiss
Mirco Weiss
Lucia Oetjen
Carolin Gehrig
Riders (retired)
Christian Egelmair
Sidney Gerber
Roger Rinderknecht
Reto Schmid
Pascal Seydoux
Rachel Seydoux
Suspension Center
Leibstadt / Zermatt / Champéry / Les Crosets
MTB Festival Leibstadt www.mtb-festival.ch
The one and only 4Cross Festival in Switzerland with multiple 4Cross races always lot of international attendees.
Swiss 4Cross Cup www.4cross.ch
The Swiss 4Cross Cup is the national 4Cross racing series. It was one of the first series with events spread over different countries like Germany and France.
Federations / Clubs
Swiss Cycling www.swisscycling.ch
VC Leibstadtwww.vcleibstadt.ch
The VC Leibstadt is one of the sucessful club in switzerland about Downhill and 4Cross. Many top riders like Samuel Zbinden or Noel Niederberger came out of this club.


text:
Ian Rapoport of the NFL Network joined Jeff Phelps and Mark Schwab to discuss Josh Gordon's off-the-field trouble, Tim Tebow news, and the possibility of Vonta Leach joining the Browns.
NFL Network's Ian Rapoport Says "Far Fewer Questions About Gordon On The Field"
OAKLAND, CA - DECEMBER 02: Josh Gordon #13 of the Cleveland Browns catches a forty-four yard touchdown pass over Ron Bartell #21 of the Oakland Raiders during the second quarter at Oakland-Alameda County Coliseum on December 2, 2012 in Oakland, California. (Photo by Thearon W. Henderson/Getty Images)


text:
The Boy and the Thug. Chase V 12/13/2010. Necessity is our strongest weapon. Moral. Character's. Thug Boy Boy Old Man.
Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.
Chase V
A boy was walking happily down the street's of Chicago to his house while a thug was following him to mug him and take his money. The boy noticed the thug following him for past few block's so he decided to run as fast as he possible could as the thug noticed his increase in speed he ran after the boy.
Soon the thug ran out of breath and slowly slowed down till he could run no more. Then old man saw the thug and said mockingly "The smallest person always run's faster than the bigger person". The thug responded with a calm voice "You do not see the difference between us I was running for his money and he was running for his life"
You There!
Then the old man pulled out a twenty dollar bill and gave it to the thug and said sarcastically " Go buy something nice". So thug changed his ways to become better person. And the boy had steak for dinner.
Have Fun!
Yeah Money!


text:
- Artist: Apocalyptica
- Format: CD
- Release Date: 4/21/2015
|your cart is empty
Apocalyptica has sold over 4 million albums internationally. Following the success of 7th Symphony, the band is set to release their 8th studio album, Shadowmaker. They have taken on a new musical challenge for the upcoming album by collaborating with Franky Perez as the exclusive singer on the vocal tracks. Shadowmaker was produced by Grammy winning rock producer Nick Raskulinecz, whose credits include Foo Fighters, Rush, Def Leppard, Paramore, Danzig, and Queens of the Stone Age, to name a few.
To play the media you will need to either update your browser to a recent version or update your version of Flash Player.


text:
Related story
Mountain West Conference
Current nine members:
- Colorado State
- Wyoming
- New Mexico
- San Diego State
- Texas Christian
- Utah
- Air Force
- Brigham Young
2011: Utah leaves for the Pac-10. BYU leaves to become independent. Boise State joins the MWC, giving the league eight members.
2012: TCU leaves for the Big East. Fresno State, Nevada and Hawaii leave the Western Athletic Conference to join the MWC, giving the MWC 10 members.
Hawaii advantages
What's good about adding Hawaii to the MWC?
- Teams that play there will be allowed to play 13 regular-season games instead of 12.
- Teams that play there will get $150,000 to $175,000 to offset travel costs.
- Hawaii has about $2.2 million in TV revenue that will be shared, SDSU President Stephen Weber said.
More on SDSU
The Mountain West Conference will consider further expansion, possibly to 11 or 12 teams, after officially adding the University of Hawaii Friday as a 10th football member for 2012, San Diego State President Stephen Weber said.
Weber serves on the league's board of directors, which is scheduled to meet in January.
"The really interesting questions are going to be what we start thinking about in January," Weber said. "The first question would be: would we like to add more programs? If so, the question is whether it's one or two (programs in addition to Hawaii). Beyond that, a question is where those programs would be. Do we want to stay in Texas, or do we want to look elsewhere?"
Weber said he doesn't expect the questions to be resolved in January. If the board is "serious about looking at a couple of teams, we'd probably do that through the rest of spring and make a decision in June," he said.
Getting to 12 teams would allow the league to split into two six-team divisions and stage a revenue-generating conference championship game.
On the other hand, adding teams could end up diluting the league's revenue shares if those new teams don't bring enough television sets and marketing value to expand the league's revenue pie.
"The league is going to analyze the situation and see if there's anything else that makes sense," SDSU Athletic Director Jim Sterk said.
In Hawaii's case, Weber said Hawaii brings $2.2 million TV revenue that now will be added to the MWC revenue-sharing pot. To offset travel costs, Sterk said Hawaii would pay $150,000 to $175,000 per trip to teams that play there.
In addition, per NCAA rules, teams that play at Hawaii are allowed to play 13 regular-season games instead of 12. So if SDSU played at Hawaii in 2012, the Aztecs could add an additional game to its schedule that year.
"Hawaii's outstanding football program and television value fit perfectly with the MWC's strategic initiatives for the future direction of the conference," MWC Commissioner Craig Thompson said in a statement.
The addition of Hawaii for 2012 comes just less than two weeks after Texas Christian announced it was leaving the MWC to play in the Big East the same season. The MWC had initiated discussions with Hawaii before TCU had decided to become the third MWC member to announce this year it was leaving the conference, including Utah, which is joining the Pac-10 next year, and Brigham Young, which is turning independent.
To fortify itself amid this changing landscape, the MWC has added four Western Athletic Conference teams. Boise State joins the league next year. In 2012, Fresno State, Nevada and now Hawaii will leave the WAC for the MWC. Hawaii will join the Big West in other sports, leaving the WAC's survival in question.
"We couldn't afford not to do it," Hawaii Athletic Director Jim Donovan said at a news conference Friday.
In effect, the MWC has traded its top three football programs for the top four football programs of the WAC. A big question is how that trade-off changes the value of the MWC's TV rights contracts, which affect each school's revenue share.
Weber said he didn't anticipate revenue shares would decrease.
"Let's be clear: the market in Utah is pretty small; that's not a big marketplace," he said. "While there would have been a market in Dallas-Fort Worth (with TCU), it wasn't being accessed (because the league's TV network, The Mtn., lacks cable distribution there)."
As for SDSU, Weber was asked if the Aztecs are committed to the MWC in the long term. "No comment," he said.
Sterk said "no offers are on the table."
Follow sports writer Brent Schrotenboer on Twitter @SchrotenboerCopyright © 2017, The San Diego Union-Tribune


text:
I like this video I don't like this video
0% (0 votes)
Duration: 6min 13sec Views: 572 Submitted: 2 years agoGet Full Access to Serious Cash - Teens Analyzed
Description: When my girlfriend told me to surprise her in bed I knew exactly she wanted to go a step further in our sex life. I've been craving to fuck her tight little butthole ever since we first had sex and my anal wishes finally came true when I slipped my erect cock into her well-oiled chocolate hole. I kept working my way deeper and deeper till my shaft completely disappeared inside of her and her moans of pleasure indicated I was on the right path to a fantastic double orgasm.
- Related Videos
- More Videos from Serious Cash - Teens Analyzed


text:
Encoxada SEÑORA CULONNA EN BUS
Description:2 TIPOS APROVECHAN EL MOMENTOY LE HACEN UNA MERIENDA CON ESTA SEÑO
Added:3 years ago
9 likes, 0 dislikes
Embed / Iframe code:
Make your selection, copy and paste the embed code above.
Standart sizes: Custom size:
What is the problem?
Please do not abuse this section. The videos ,which are on investigate list, are manually checked by editor.
Security code:(is not case-sensitive) can't read
Write a comment
at least 10 characters, up to 400 characters
400 characters remaining
Security code:
(is not case-sensitive) can't read


text:
|Home | About | Journals | Submit | Contact Us | Français
We have identified Rabconnectin-3 alpha and beta (Rbcn-3A and B) as two regulators of Notch signaling in Drosophila. We found that, in addition to disrupting Notch signaling, mutations in Rbcn-3A and B cause defects in endocytic trafficking, where Notch and other membrane proteins accumulate in late endosomal compartments. We show that Notch is transported to the surface of mutant cells and that signaling is disrupted after the S2 cleavage. Interestingly, the yeast homolog of Rbcn-3A, Rav1, regulates the V-ATPase proton pump responsible for acidifying intracellular organelles. We found that, similarly, Rbcn-3A and B appear to regulate V-ATPase function. Moreover, we identified mutants in VhaAC39, a V-ATPase subunit, and showed that they phenocopy Rbcn-3A and Rbcn-3B mutants. Our results demonstrate that Rbcn-3 affects Notch signaling and trafficking through regulating V-ATPase function, which implies that the acidification of an intracellular compartment in the receiving cells is crucial for signaling.
Notch signaling is a highly conserved cell communication pathway widely used in animal species (Artavanis-Tsakonas et al., 1999). Aberrant Notch signaling is associated with developmental disorders and cancers in humans (Lai, 2004; Talora et al., 2008). The core components of the Notch pathway are the single-pass transmembrane receptor Notch, its ligands and the Suppressor of Hairless transcription factor. Signaling is initiated upon ligand binding to Notch. This triggers two consecutive proteolytic cleavage events; a first extracellular cleavage mediated by ADAM-family metalloproteases, followed by an intramembranous cleavage by γ-secretase. As a result, the intracellular domain of Notch (NICD) is released and translocates into the nucleus to regulate transcription (Struhl et al., 1993; Rebay et al., 1993; Lai, 2004; Schweisguth, 2004).
The activity of both Notch and its ligands relies strongly on posttranslational modifications and intracellular trafficking (Le Borgne, 2006; Nichols et al., 2007; Stanley, 2007). Notch ligand activity requires the endocytic machinery. Similarly, Notch activation involves endocytic trafficking, but the precise mechanism by which endocytosis of Notch contributes to signaling activity remains unresolved. In Drosophila, mutations that block Notch trafficking at different endocytic steps have distinct effects on Notch signaling activity (Childress et al., 2006; Gallagher and Knoblich, 2006; Jaekel and Klein, 2006; Kanwar and Fortini, 2008; Moberg et al., 2005; Rusten et al., 2006; Seugnet et al., 1997; Thompson et al., 2005; Vaccari and Bilder, 2005; Vaccari et al., 2008). To explain the relation between Notch endocytic trafficking and Notch signaling activity, it has been proposed that Notch entry into early endosomes is critical for γ-secretase mediated Notch cleavage and thus normal Notch signaling, and that retention in the early endosome causes increased Notch signaling (Le Borgne, 2006; Nichols et al., 2007; Vaccari et al., 2008).
Here, we report the identification of the Drosophila homologues of mammalian Rabconnectin-3 alpha and beta (Rbcn-3A and B) and show that they are required for Notch signaling and endocytic trafficking in follicle cells and imaginal disc cells. In the absence of Rbcn-3A and B, Notch and other membrane proteins accumulate in an aberrant late endosomal compartment. Interestingly, the yeast homologue of Rbcn-3A, Rav1, was shown to regulate the assembly and activity of the vacuolar (H+) ATPase (V-ATPase) (Seol et al., 2001; Smardon et al., 2002). V-ATPases are ATP-driven proton pumps composed of two multi-subunit complexes: the membrane V0 complex and the peripheral V1 complex (Forgac, 2007; Jefferies et al., 2008). They are responsible for the acidification of intracellular compartments and have a well-established role in protein sorting, trafficking and turnover (Forgac, 2007; Jefferies et al., 2008). We show that the endocytic defects we observe in Rbcn-3A and B mutants are consistent with Rbcn-3 regulating V-ATPase activity. Moreover, we identify mutants in the V0 subunit VhaAC39 and show that their phenotypes with respect to Notch signaling and protein trafficking are identical to those in Rbcn-3 mutants. Our results indicate that Rbcn-3 acts primarily through regulating V-ATPase function and thus reveal a functional connection between the vacuolar proton pump and Notch signaling.
In a genetic mosaic screen aimed at identifying genes regulating the organization and morphogenesis of the Drosophila follicular epithelium (Denef et al., 2008), we isolated two complementation groups which caused defects reminiscent of those observed in follicle cells mutant for Notch. The two complementation groups were initially named after the representative alleles FK39 and FV10 and contained 13 and 17 alleles respectively.
Wild-type follicle cells divide mitotically from stage 2 to stage 6 of oogenesis and then switch from a mitotic cycle to an endocycle. From stage 7 to 10A follicle cells go through three rounds of DNA duplication without cell division (Horne-Badovinac and Bilder, 2005). This mitotic cycle to endocycle switch is regulated by the Notch pathway. Follicle cells in which Notch signaling is compromised keep dividing after stage 6 and fail to undergo endoreplication. As a result, follicle cell clones mutant for Notch form patches of cells with smaller nuclei than neighboring wild-type cells (Deng et al., 2001; Lopez-Schier and St Johnston, 2001). Follicle cell clones mutant for FK39 and FV10 show a similar phenotype (Figure 1A and data not shown). To confirm that the mutant cells continue dividing after stage 6 we stained for the mitotic marker phosphorylated Histone H3 (PH3) and found that, in contrast to wild-type egg chambers where PH3 positive cells are only found up to stage 6, PH3 staining was frequently present after stage 6 in these mutant cells (Figure 1B and data not shown).
The failure of FK39 and FV10 mutant follicle cells to switch from a mitotic cycle to an endocycle suggests defective Notch signaling. To test this more directly, we examined the expression of several known Notch targets in the follicular epithelium. The transcription factor Hindsight (Hnt) is upregulated by Notch signaling at stage 6/7 and this upregulation is required for the mitotic-cycle-to-endocycle switch in follicle cells (Sun and Deng, 2007). In contrast to wild-type cells Hnt upregulation was not detected in FK39 and FV10 mutant follicle cells in egg chambers beyond stage 6 (Figure 1C and data not shown). The homeodomain protein Cut is downregulated in wild-type follicle cells upon Notch activation at stage 6 and this downregulation is critical for follicle cells to enter the endocycle (Sun and Deng, 2005). In FK39 and FV10 mutant follicle cells Cut expression fails to be repressed after stage 6 (Figure 1D and data not shown). Finally, we examined the expression of a transcriptional reporter for Notch signaling, Gbe+Su(H)-lacZ (Furriols and Bray, 2001). In 77% of FK39 mutant clones (n=84), β-galactosidase staining was either absent or strongly reduced (Figure 1E). Taken together, these results indicate that Notch signaling is lost in FK39 and FV10 mutant follicle cells.
To ask whether the gene products disrupted in these mutants were required specifically in the follicular epithelium for Notch signaling or whether they also affected Notch signaling in other tissues, we examined the expression of the Gbe+Su(H)-lacZ reporter in FK39 mutant eye disc cells. Similar to our results in the follicle cells, we detected much lower levels of β-galactosidase in FK39 mutant eye disc cells compared to neighboring wild-type cells (Figure 1F, n>30), demonstrating that Notch signaling is compromised in FK39 mutant eye disc cells.
The observed loss of Notch signaling in FK39 and FV10 mutant follicle cells indicates that the gene products disrupted in FK39 and FV10 are required for Notch signaling in the signal receiving cells. Indeed, it was shown that the Notch ligand Delta is required in the germ line but not in the follicle cells to induce the mitotic to endocycle switch (Deng et al., 2001; Lopez-Schier and St Johnston, 2001). Upon ligand binding, Notch undergoes an extracellular cleavage (S2), generating the membrane bound NEXT, followed by an intramembranous cleavage (S3) producing NICD (Lai, 2004; Schweisguth, 2004). To further define the step where Notch signaling is disrupted in the mutants, we overexpressed full length Notch, NEXT or NICD constructs in FK39 mutant clones, using the MARCM system (Lee and Luo, 1999). With this system of clone generation, Hindsight failed to be upregulated in 83% of FK39 follicle cell clones (n=180) in stage 6/7 egg chambers, again indicating defective Notch signaling. Disrupted Notch signaling was similarly observed in 83% of FK39 clones with coexpression of full length Notch (n=121), and in 71% of FK39 clones which expressed NEXT (n=139), but only in 10% of FK39 clones which expressed NICD (n=122). (Figure 2A-D) Therefore, only the expression of NICD but not full length Notch or NEXT, can restore the loss of Notch signaling in FK39 mutant follicle cells.
In wing imaginal discs, overexpression of NEXT results in ectopic activation of the Notch target gene Cut (Rebay et al., 1993; Struhl and Greenwald, 1999). We therefore compared wing discs expressing NEXT in FK39 mutant clones with discs expressing NEXT in control clones. Cut was ectopically expressed in all cells of 59% of control clones (n=172) but only in 33% of FK39 mutant clones (n=152) (p<10-3, binomial probability) (Figure 2E, F). It is possible that perdurance of the wild-type gene product between the time of clone induction and dissection of the discs prevented a completely penetrant loss of signaling activity. Nevertheless, this result indicates that NEXT signals less efficiently in FK39 mutant cells than in control cells. We also attempted to assess whether ectopic Notch signaling induced by overexpession of NICD in wing discs could be suppressed by the loss of the FK39 gene product. However, we were unable to obtain a significant number of surviving FK39 clones expressing NICD in this experiment.
Taking together the evidence in both follicle cells and wing disc cells, we conclude that the gene product disrupted in FK39 mutant cells is required for signaling after the S2 cleavage of Notch but does not significantly affect Notch signaling after the release of the intracellular domain into the cytoplasm.
Using meiotic recombination with visible recessive markers and P element insertions we mapped the lethal phenotype of FK39 and FV10 to the chromosomal regions 2B15 and 5E respectively. The fact that these two complementation groups were mapped to distinct loci, yet shared identical phenotypes suggested that the two gene products might be functionally related. Two ORFs in these regions therefore stood out; CG3585 in region 5E and CG17766 in region 2B15 encode the fly homologues of the mammalian WD-40 proteins Rabconnectin-3 alpha and beta (Rbcn-3A and B) respectively. Rbcn-3A and B were identified as binding partners of Rab3-GEP and Rab3-GAP in synaptic vesicle extracts of rat brain (Kawabe et al., 2003; Nagano et al., 2002; Sakisaka and Takai, 2005). The two proteins appear to form a stable complex but the biochemical function of this complex remains unknown. The Drosophila Rbcn-3A gene was originally named DmX (Kraemer et al., 1998) and its mammalian homologues Dmx-like1 and Dmx-like2, with Rbcn-3A being Dmx-like2 (Kraemer et al., 2000; Nagano et al., 2002). Because of the functional connection between Rbcn-3A (DmX) and Rbcn-3B we will use the name Rbcn-3A in this paper.
To ask whether the loss of Rbcn-3A and B was indeed responsible for the phenotypes observed in FK39 and FV10 mutants, we sequenced the Rbcn-3B gene in seven alleles of the FK39 group and found that all of them contained mutations in Rbcn-3B (Figure 3A). Moreover, expression of a HA-tagged Rbcn-3B cDNA in FK39 mutant follicle cell clones rescued the Notch-like phenotype (Figures 3B and C, n>50). Similarly, a genomic rescue construct containing the Rbcn-3A gene and ~1.5 kb upstream and downstream sequences rescued the lethality and ovarian phenotypes of FV10 mutants (Figure 3D).
Taken together, our results demonstrate that the loss of Notch signaling observed in FV10 and FK39 mutants is caused by the loss of function of the Drosophila homologues of Rbcn-3A and Rbcn-3B respectively. Since the two genes share the same mutant phenotypes in all aspects of our analysis we will refer to them collectively as Rbcn-3, where appropriate.
We have shown that Rbcn-3 is required for Notch signaling in the receiving cells. Since effective Notch signaling relies on intracellular trafficking of Notch in the signal receiving cells (Le Borgne, 2006; Nichols et al., 2007) we asked whether Notch trafficking was defective in Rbcn-3 mutant cells. We therefore compared the subcellular distribution of Notch in Rbcn-3 mutant follicle cells and eye disc cells with that in wild-type cells. We found that in Rbcn-3 mutant follicle cells as well as eye disc cells, Notch accumulated in enlarged intracellular compartments (Figures 4A-E and data not shown). Notch accumulation in Rbcn-3 mutant cells was observed with antibodies against both the intracellular domain (NICD, Figures 4A, B and D) and the extracellular domain (NECD, Figures 4C and E), suggesting that full length Notch is present in these enlarged compartments.
We next asked whether the loss of Rbcn-3 specifically disrupts the distribution of Notch or whether other transmembrane proteins were also affected. We examined the subcellular localization of Delta (Figures 4F, G), EGF receptor (Supplemental Figures 1A, B), Domeless (Supplemental Figure 1C), Fas II (Figures 4H, I), FasIII and DE-Cadherin (data not shown) and found that they all accumulate in similar enlarged compartments in the absence of Rbcn-3. In contrast, membrane-associated proteins, such as aPKC, Crag and Dlg maintained a normal staining pattern in Rbcn-3 mutant follicle cells (Figures 4J, K and data not shown). Interestingly, despite the intracellular accumulation of the EGF receptor and of Domeless, which activates the JAK/STAT pathway, signaling downstream of these two receptors occurs normally in Rbcn-3 mutant follicle cells (Supplemental Figure 1).
The intracellular accumulation of Notch and other transmembrane proteins in Rbcn-3 mutant cells clearly demonstrates a role for Rbcn-3 in regulating protein trafficking. To ask whether the accumulation of membrane proteins in Rbcn-3 mutant cells is due to a defect in endocytic or exocytic trafficking, we performed a Notch trafficking assay in eye discs (Le Borgne and Schweisguth, 2003; Lu and Bilder, 2005). Rbcn-3B mosaic eye discs were incubated in cold medium containing an antibody against NECD to specifically label Notch at the cell surface. Pulse-labeled discs were either fixed immediately to examine the amount of Notch at the cell surface or incubated for different times in fresh medium at 25°C prior to fixation to follow endocytosis of labeled Notch. We did not observe a significant difference in the amount of Notch at the cell surface between wild-type and Rbcn-3B mutant cells, indicating that exocytosis of Notch is not noticeably affected in the absence of Rbcn-3B (Figure 5A and data not shown). Significantly, after a 1.5 h chase, abnormal accumulation of internalized Notch with bound antibody was observed in Rbcn-3B mutant cells compared to neighboring wild-type cells (Figure 5B). These results indicate that, in the absence of Rbcn-3, Notch reaches the surface of eye disc cells, is internalized, but then accumulates in an endocytic compartment.
A similar trafficking experiment is technically not possible in the follicle cells. To address whether Notch accumulation in Rbcn-3 mutant follicle cells results from defects in endocytic trafficking, as it does in eye disc cells, we compared Notch localization in Rbcn-3B mutant follicle cells with that in cells mutant for both Rbcn-3B and either Clathrin heavy chain (Chc) or shibire (shi), the fly orthologue of dynamin. Both Chc and Shi are required for the formation of endocytic vesicles at the plasma membrane (Ungewickell and Hinrichsen, 2007). In Chc and shi mutant follicle cells, Notch accumulates at the cell surface (Figure 5C, n>50 and data not shown)(Lu and Bilder, 2005; Vaccari et al., 2008). As described earlier, in Rbcn-3 mutant follicle cells Notch accumulates in enlarged intracellular compartments (e.g. Figures 4A-C, n>100). By contrast, in Rbcn-3B Chc and Rbcn-3B shi double mutant follicle cells Notch accumulated around the cell periphery as in Chc or shi single mutant follicle cells (Figure 5D, n>50 and data not shown). These results indicate that Rbcn-3 is not required for Notch to reach the cell surface and that the intracellular accumulation of Notch in the absence of Rbcn-3 requires Chc and Shi-dependent internalization.
Taken together, our results demonstrate that the major defect in Notch trafficking in Rbcn-3 mutant follicle cells and eye disc cells occurs in the endocytic pathway and that exocytosis occurs normally.
To determine the nature of the endocytic compartment in which Notch accumulates in the absence of Rbcn-3, we examined the distribution of known endocytic markers in Rbcn-3B mutant follicle cells and found that the enlarged compartments in mutant cells were positively labeled with the late endosomal marker YFP-Rab7 (Figure 5E) (Zhang et al, 2007), but not with the early endosomal markers GFP-Rab5 (data not shown) or GFP-2xFYVE (Figure 5F)(Wucherpfennig et al., 2003) or the recycling endosomal marker GFP-Rab11 (data not shown)(Emery et al., 2005). These results indicate that Notch and other membrane proteins accumulate in an enlarged late endosomal compartment in the absence of Rbcn-3.
To further characterize the nature of this late endosomal compartment, we stained egg chambers with an antibody detecting ubiquitinated proteins. Ubiquitination of membrane proteins often serves as a signal for their internalization and endocytic sorting prior to lysosomal degradation (Raiborg et al., 2003). In cells lacking components of the endosomal sorting complexes required for transport (ESCRTs), such as Hrs, Vps25 and Tsg101, which mediate protein sorting in the endocytic pathway, ubiquitinated proteins strongly accumulate in enlarged endosomes (Figure 5G) (Jekely and Rorth, 2003; Moberg et al., 2005; Thompson et al., 2005; Vaccari and Bilder, 2005). In contrast, the enlarged endosomes containing Notch and other membrane proteins in Rbcn-3B mutant cells do not contain increased levels of ubiquitin (Figure 5H).
Our data indicate that the loss of Rbcn-3 in Drosophila follicle cells causes membrane proteins to accumulate in enlarged late endosomes after deubiquitination has taken place. Intriguingly, other mutants that cause Notch accumulation in late endosomal compartments, such as the phosphatidylinositol-5-kinase Fab1, and the HOPS complex components carnation and deep orange, have no effect on Notch signaling (Rusten et al., 2006; Sevrioukov et al., 1999; Sriram et al., 2003 and data not shown). The accumulation of Notch in these enlarged late endosomal compartments in Rbcn-3 mutant cells can therefore not account for the observed loss of Notch signaling.
We have shown two major defects in Rbcn-3 mutant cells: a loss of Notch signaling and an accumulation of Notch and other membrane proteins in abnormal late endosomal compartments. Given that the late trafficking defect cannot explain the block in Notch signaling, we wanted to determine the underlying common cellular defect that could lead to these phenotypes. Upon closer inspection of the Rbcn-3A protein sequence we noticed that it shares homology with the yeast protein Rav1(Seol et al., 2001; Sipos et al., 2004), as well as the C. elegans Rav1 ortholog rbc-1 (wormbase). Rav1 is a component of the RAVE complex (regulator of the (H+)-ATPase of the vacuolar and endosomal membranes), which regulates the activity of the vacuolar (H+) ATPase (V-ATPase) (Seol et al., 2001; Smardon et al., 2002). The V-ATPases are conserved ATP-driven proton pumps composed of two multi-component subcomplexes, the membrane bound V0, which mediates proton translocation, and the peripheral V1, responsible for ATP hydrolysis (Forgac, 2007; Jefferies et al., 2008). The yeast RAVE complex promotes the assembly of V1 and V0 to form the active holo-enzyme (Seol et al., 2001; Smardon et al., 2002). One of the major functions of V-ATPases is to acidify the lumens of various intracellular organelles, including endosomes, lysosomes and secretory vesicles (Forgac, 2007; Jefferies et al., 2008). To test whether Rbcn-3A is required for this V-ATPase function in Drosophila as Rav1 is in yeast, we monitored the acidity of intracellular compartments with the pH-sensitive vital dye LysoTracker. In wild-type follicle cells acidic organelles were evident by intense LysoTracker staining (Figure 6A, B). By contrast, Rbcn-3A and Rbcn3-B mutant follicle cells showed strongly reduced or no LysoTracker staining (Figure 6A, B). This observation indicates a lack of acidic compartments in the absence of Rbcn-3 and is consistent with a role for Rbcn-3 in regulating V-ATPase-dependent acidification. In yeast, the RAVE complex was shown to bind the V1 subcomplex of the V-ATPase (Seol et al., 2001; Smardon et al., 2002). We therefore asked whether Rbcn-3 interacts with components of the V1 subcomplex in Drosophila. We immuno-precipitated two Drosophila V1 subunits, Vha55 (V1 B subunit) and VhaSFD (V1 H subunit) from ovaries expressing HA-tagged Rbcn-3B in follicle cells and found that HA-Rbcn-3B co-precipitated with both subunits (Figure 6C and data not shown). This result further confirms that Rbcn-3 could serve to regulate V-ATPase activity in Drosophila.
If Rbcn-3 indeed affects both Notch signaling and protein trafficking through regulating V-ATPase function, mutations in V-ATPase subunits should show similar defects as Rbcn-3 mutants. In the genetic screen that lead to the identification of Rbcn-3A and B, we found two additional mutants, FZ29 and FY38, that resulted in mosaic patches of cells with small nuclei. Upon closer inspection, we found that these mutants showed identical phenotypes to Rbcn-3 mutants. We mapped the lethality of these two mutants between two transposable elements at 3D4 and 4C2, and very close to the recessive marker echinus at 3F2 (Figure 7A). Interestingly, a single V-ATPase subunit gene, VhaAC39, maps exactly within this region (3F4) (Allan et al., 2005). We therefore sequenced the VhaAC39 gene in FZ29 and FY38, and found that, in both alleles, part of the VhaAC39 ORF was deleted. FY38 contains a 385 bp deletion combined with a 2 bp insertion and FZ29 contains a 137 bp deletion leading to a frame shift and a premature stop codon (Figure 7A and Supplemental Figure 2). VhaAC39 encodes one of the two Drosophila V0 d subunits (Allan et al., 2005; Nishi et al., 2003; Smith et al., 2002). As expected from the loss of VhaAC39 activity and thus V-ATPase function, LysoTracker staining was lost in VhaC39 mutant follicle cells (Figure 7B). In addition, Notch and other membrane proteins accumulated in enlarged intracellular compartments that were labeled strongly with the late endosomal marker GFP-Rab7 but not with the early endosomal markers GFP-Rab5, 2xFYVE-GFP, the recycling endosomal marker GFP-Rab11 or the Golgi marker Lava lamp (Figures 7C-F and data not shown). Very significantly, we found that Notch signaling was lost in the absence of VhaAC39; VhaAC39 mutant follicle cells keep dividing after stage 6, fail to upregulate Hnt, fail to downregulate Cut and show strongly reduced levels of Gbe+Su(H)-lacZ (Figure 8). Finally, VhaAC39 mutant eye discs cells behaved similarly to Rbcn-3 mutant eye disc cells in the Notch trafficking assay; Notch reached the surface, was internalized and accumulated in intracellular compartments (data not shown).
In summary, our results demonstrate that mutations in the V0 subunit VhaAC39 phenocopy mutations in Rbcn-3 with respect to protein trafficking and Notch signaling. We can therefore conclude that Rbcn-3 acts primarily through its effect on V-ATPase activity to regulate Notch signaling and endocytic trafficking. Moreover, our results provide evidence for a role of the vacuolar proton pump in the regulation of Notch signaling.
In a genetic screen in the follicular epithelium, we have identified mutations in the fly homologues of the WD40 proteins Rbcn-3A and B, and a V-ATPase V0 d subunit, VhaAC39, and have shown that interfering with V-ATPase function leads to a block in Notch signaling in Drosophila.
Mammalian Rbcn-3A and B were shown to form a complex that interacts with both Rab3-GAP and Rab3-GEF but the biological function of this complex had remained elusive (Kawabe et al., 2003; Nagano et al., 2002; Sakisaka and Takai, 2005). Interestingly, the closest homologue of Rbcn-3A in yeast is the protein Rav1, a component of the RAVE complex (Seol et al., 2001; Sipos et al., 2004). The RAVE complex was shown to interact with the V1 subcomplex of the V-ATPase and promote its activity by regulating the assembly of the peripheral V1 and membrane V0 subcomplexes to form the V-ATPase holoenzyme (Seol et al., 2001; Smardon et al., 2002). V-ATPases are evolutionarily conserved ATP driven proton pumps responsible for the acidification of several intracellular compartments, including endosomes, lysosomes, secretory vesicles and the Golgi apparatus (Forgac, 2007; Jefferies et al., 2008). Inhibition of V-ATPase function leads to a failure in luminal acidification of these different compartments. Likewise, in Rav1 mutants in yeast, V-ATPase dependent vacuolar acidification is disrupted (Seol et al., 2001). We have shown that, similarly, Rbcn-3A and B mutant follicle cells in Drosophila fail to acidify intracellular compartments. We showed a comparable lack of acidic compartments in follicle cells mutant for the VhaAC39 gene, which encodes one of the two V-ATPase V0 d subunits (Allan et al., 2005; Nishi et al., 2003; Smith et al., 2002). The V0 d subunit was suggested to regulate the coupling of ATP hydrolysis and proton translocation and is therefore indispensable for V-ATPase activity (Nishi et al., 2003; Smith et al., 2008).
In eukaryotic cells, V-ATPase dependent acidification of organelles is necessary for protein sorting, trafficking and turnover (Forgac, 2007; Jefferies et al., 2008; Nelson, 2003). For instance, hydrolases responsible for protein degradation in the lysosome have an optimal activity at a low pH. In addition, several trafficking steps along the endocytic pathway have been shown to rely on V-ATPase function (Forgac, 2007; Marshansky and Futai, 2008). Mutations in V-ATPase components or pharmacological inhibition of V-ATPase activity result in an accumulation of membrane proteins in endocytic compartments and in some cases block transport between the late endosome and the lysosome (Nelson, 2003; van Deurs et al., 1996; van Weert et al., 1995). Likewise, Rav1 mutants in yeast show an accumulation of endosomes and a delay in vacuolar transport and degradation (Sipos et al., 2004). Consistent with these data, we have shown that in Rbcn-3 mutant cells Notch and other integral membrane proteins accumulate in enlarged late endocytic compartments. We observed an identical phenotype upon disruption of VhaAC39.
The RAVE complex was identified in yeast but so far a similar complex has not been described in higher eukaryotes. By demonstrating a striking resemblance between Rbcn-3 and VhaAC39 mutant cells with respect to intracellular acidification and protein trafficking we have provided evidence for the existence of a similar complex in higher eukaryotes. This conclusion is supported by our observation that HA-tagged Rbcn-3B can be immuno-precipitated with at least two components of the V1 subcomplex, the B subunit Vha55 and the H subunit VhaSFD. In addition to Rav1the yeast RAVE complex contains two other components: Rav2 and Skp1. Skp1 is a highly conserved SCF ubiquitin ligase that forms multiple distinct complexes involved in a wide array of cellular processes. Rav2 on the other hand has no obvious homologues in Drosophila or other higher eukaryotes. Conversely, no clear Rbcn3-B homologue exists in yeast. In rat, the Rav1 homologue Rbcn-3A forms a complex with Rbcn3-B (Kawabe et al., 2003) and based on the identical phenotypes of Rbcn-3A and B mutants in Drosophila they likely act in a complex in flies as well. An interesting possibility is therefore that Rbcn-3B performs the function of Rav2 in the Drosophila RAVE complex.
Interestingly, we isolated Rbcn-3 and VhaAC39 mutants in our screen because of their phenotypic similarity to mutations in Notch pathway components and confirmed that indeed both Rbcn-3 and VhaAC39 are critical factors required for Notch signaling. During Drosophila oogenesis Notch signaling is required for multiple processes. In particular, at stage 6 of oogenesis, Delta, expressed in the germline signals to Notch in the follicle cells to initiate the switch from mitosis to endocycle (Deng et al., 2001; Lopez-Schier and St Johnston, 2001). We have shown that the loss of Rbcn-3 or VhaAC39 in follicle cells phenocopies defective Notch signaling with respect to the mitosis-endocycle switch. In addition, we observed defects in other Notch–dependent processes during oogenesis in the absence of Rbcn-3 or VhaAC39, including fused egg chambers and anterior-posterior polarity defects and found that Rbcn-3 also affects Notch signaling in eye discs. Our results showing that disrupting either a V-ATPase subunit (VhaAC39) or a V-ATPase regulator (Rbcn-3) both lead to a loss of Notch signaling provides evidence for a role of the vacuolar proton pump in the regulation of Notch signaling.
How could the vacuolar proton pump regulate Notch signaling? Since the loss of Notch signaling is evident upon disruption of Rbcn-3 or VhaAC39 function in the follicle cells, the signal receiving cells with respect to Notch signaling, it is clear that V-ATPase function must be required at the level of Notch or in a downstream signaling event. We have shown that, in the absence of Rbcn-3 or VhaAC39, Notch accumulates strongly in enlarged late endosomal compartments, consistent with the known role for V-ATPases in endocytic trafficking and lysosomal degradation. However, it is highly unlikely that the accumulation of Notch in this late endosomal compartment is responsible for the observed block in signaling. Defects in early endocytic trafficking of Notch have been correlated to aberrant Notch signaling. Mutations in proteins that affect the first steps in endocytosis, such as Chc, the fly dynamin Shi, the Rab5 GTPase and the syntaxin Avalanche lead to a loss of Notch activity (Seugnet et al., 1997; Vaccari et al., 2008). In these mutants Notch accumulates at the cell surface and fails to reach the early endosome (Vaccari et al., 2008). In contrast, mutations in some proteins that affect sorting at the MVB, such as the endosomal sorting complex required for transport (ESCRT) components Tsg101 and Vps25, cause ectopic activation of the Notch pathway. In these mutants Notch accumulates with ubiquitinated cargo in enlarged MVB (Moberg et al., 2005; Thompson et al., 2005; Vaccari and Bilder, 2005; Vaccari et al., 2008). However, mutations in proteins acting at the late endosome, such as the Phosphatidylinositol 3-Phosphate 5-kinase Fab1, cause Notch to accumulate in the late endosome after deubiquitination has occurred (Rusten et al., 2006). Significantly, whereas these late endosomal mutants show Notch accumulation in late endosomal compartments comparable to that seen in Rbcn-3 and VhaAC39 mutants, they do not perturb Notch signaling (Rusten et al., 2006; Vaccari et al., 2008). This indicates that the accumulation of Notch in late endosomes upon disruption of V-ATPAse activity cannot explain the loss of Notch signaling.
In addition to a role in endocytosis and lysosomal degradation, V-ATPases function in the secretory pathway, both at the level of protein sorting in the Golgi and in the fusion of secretory vesicles with the plasma membrane (Forgac, 2007; Marshansky and Futai, 2008). It has also been observed that acidification of vesicles is important for recruiting certain cytosolic coat proteins (Gu and Gruenberg, 2000). It was therefore possible that the loss of Notch signaling in Rbcn-3 or VhaAC39 mutant cells was caused by a defect in the trafficking of Notch to the cell surface. We found no evidence for a requirement of Rbcn-3 or VhaAC39 in exocytosis and showed that Notch and other membrane proteins still reach the cell surface in the absence of Rbcn-3 or VhaAC39. Nevertheless, it remains possible that VhaAC39 or Rbcn-3 mutants show subtle defects in exocytic trafficking or in the posttranslational modification of Notch in the secretory pathway, which may then be responsible for a loss of signaling.
It is possible that the loss of Notch signaling in VhaAC39 and Rbcn-3 mutant cells is due to a defect in the trafficking and/or activity of another pathway component. A possible candidate is γ-secretase. γ-secretase-mediated S3 cleavage of NEXT, the Notch product generated upon ligand-induced cleavage by ADAM metalloproteases, results in the generation of the active form of Notch, NICD (Lai, 2004; Schweisguth, 2004). We have shown that the expression of NICD but neither full length Notch nor NEXT can rescue defective Notch signaling in Rbcn-3 mutant follicle cells. Furthermore, we have shown that NEXT ectopically expressed in wing imaginal discs signals much less efficiently in the absence of Rbcn-3B. These results are consistent with a requirement for V-ATPase activity at the level of or downstream of the S3 cleavage. The γ-secretase complex consists of four core transmembrane proteins (Dries and Yu, 2008). The complex is assembled in the ER and shuttles between the ER and the Golgi. A small fraction of the γ-secretase complex is transported to the plasma membrane and endosomes where it is thought to mediate the cleavage of its substrates such as NEXT (Dries and Yu, 2008). It is conceivable that disrupting V-ATPase activity results in the aberrant trafficking of the γ-secretase complex, thus preventing the S3 cleavage of Notch.
Alternatively, it has been reported that γ-secretase activity is optimal at a low pH (Knops et al., 1995; Pasternak et al., 2004; Schrader-Fischer and Paganetti, 1996; Siman et al., 1993). Recent evidence also indicates that S3 cleavage of Notch can generate heterogenous fragments that differ by a few amino acids with different stabilities thus exhibiting different signaling potencies (Tagami et al, 2008). Therefore, a loss of V-ATPase activity and the resulting alkalization of intracellular organelles could affect the generation or the release of the S3 cleavage product. In addition, a recent study suggested that mutations in the Aquaporin Big brain, which is required for Notch signaling (Campos-Ortega and Knust, 1990), also show a reduced luminal acidification (Kanwar and Fortini, 2008).
In summary, our results have demonstrated that regulating V-ATPase activity is fundamental to Notch signaling in Drosophila.
Rabconnectin-3A, Rabconnectin-3B and VhaAC39 mutations were isolated in a mosaic screen described by Denef et al. (Denef et al., 2008). In this screen, mutations were induced by EMS in y w FRT19 flies. The ChcGF23 and shiFL54 alleles used in the double mutant analysis were isolated in the same screen (N. D., Y. Y. and T. S., unpublished) and validated through complementation tests with the Chc1 and shiDN alleles (Bloomington). FRT40A hrsD28 flies were a gift from H. Bellen (Lloyd et al., 2002). Duplication and P-element lines used for mapping were obtained from Bloomington. Reporter lines used to assay signaling pathway activity are Gbe+Su(H)-lacZ (Furriols and Bray, 2001), mirror-lacZ (Xi et al., 2003) and kekkon-lacZ (Pai et al, 2000). The UAS-Notch line was a gift from G. Struhl. UAS-NEXT and UAS-NICD lines were gifts from S. Artavanis-Tsakonas (Rebay et al., 1993). Transgenic lines used to mark endocytic compartments are GFP-Rab5 (Wucherpfennig et al., 2003), GFP-Rab7 (Entchev et al., 2000), YFP-Rab7 (Zhang et al., 2007), GFP-Rab11 (Emery et al., 2005) and GFP-2xFYVE (Wucherpfennig et al., 2003). Follicle cell clones were generated using the FRT/UAS-Flp/GAL4 system (Duffy et al., 1998). Eye disc clones were generated using FRT/eyFlp. Follicle cell clones and wing disc clones which also express Notch constructs were generated using the MARCM system (Lee and Luo, 1999). The genotypes of flies and larvae used for the analyses are described in the Supplemental Data.
Ovaries and eye discs were dissected, fixed and stained by standard procedures. The Notch endocytic trafficking assay was conducted by following the procedure described in (Le Borgne and Schweisguth, 2003). For LysoTracker staining, ovaries were dissected, incubated in M3 culture medium containing 1μM LysoTracker Red DND-99 (Molecular Probes) for 5 min, rinsed three times with M3 medium and imaged immediately. Primary antibodies used are listed in the supplemental data. AlexaFluor 568 and 647 conjugated secondary antibodies and Hoechst were from Molecular Probes. Images were taken on a Zeiss LSM510 or a Leica SP5 confocal microscope.
Immuno-precipitation (IP) and western blotting was conducted by following the procedure described in (Pane et al., 2007). The antibodies used for IP were rabbit anti-vha55 (1: 500, (Du et al., 2006)) and anti-vhaSFD (1:500, (Pyza et al., 2004)). Both antibodies are kind gifts from J. A. Dow. The negative control was a non-specific rabbit pre-immune serum. The antibody used for western blotting was rat anti-HA-Peroxidase (1:3000, Roche).
Meiotic recombination mapping with visible recessive markers placed the lethal mutation in FV10 between crossveinless (5A13) and cut (7B4). It was further mapped between the P-elements BG02156 (5E1) and KG00403 (5E4). FK39 mutants were mapped using Duplication and Deficiency mapping. The lethality of FK39 mutants was rescued by Dp(1;Y)67g and complemented by the Deficiency lines Df(1)AD11, Df(1)S39, Df(1)6443, Df(1)Exel8196, Df(1)Pgd35 and Df(1)ED409 and by exclusion thus mapped to 2B15. PiggyBacs used for mapping of the lethal FZ29 and FY38 mutations were PBac(WH)CG16782f06392 (3D4) and PBac(WH)CG3626f02285 (4C2). To identify the molecular lesions in Rbcn-3B and VhaAC39 alleles, the mutant lines were balanced over an FM7, Kr>GFP balancer chromosome and genomic DNA from GFP- larvae (for Rbcn-3B) or embryos (for VhaAC39) was isolated. PCR products covering the Rbcn-3B or VhaAC39 gene regions of two independent genomic preps were sequenced and sequences were compared to FRT19 control sequences.
To make the Rbcn-3A genomic rescue construct, the Rbcn-3A gene and 1.5 kb upstream and downstream sequences were cloned from the BAC clone RP98-19D3 by recombineering cloning (Venken et al., 2006). To generate the pUASp-Rbcn-3B-HA construct, the coding region of Rbcn-3B (CG17766), which is contained in a single exon, was PCR amplified from the BAC clone RP98-5N9 and cloned into pUASp in frame with a C-terminal HA tag.
We thank H. Bellen, S. Bray, G. Struhl, S. Artavanis-Tsakonas, H. Krämer, S. Noselli, D. Harrison, M. Metztein, M. Gonzalez-Gaitan, JA. Dow, the Developmental Studies Hybridoma Bank and the Bloomington stock center for providing flies and antibodies; G. Barcelo for technique help; J. Goodhouse for advice with confocal microscopy and members of the Schüpbach and Wieschaus labs for feedback and advice. We also thank S. De Renzis and E. Wieschaus for helpful comments on the manuscript. This work was supported by the Howard Hughes Medical Institute and US Public Health Service Grant RO1 GM077620.
Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


text:
TOPEAK Bike Stand PrepStand Elite
Article details
TOPEAK stand PrepStand Elite
Professional Repair Stand Topeak! A stable Dreibeinfuß for stability. Using the finely adjustable and 360 ° rotatable clip can be found for every situation the right position of the bicycle.
Compared to PrepStand Pro a small parts tray is fitted instead of the digital scale in this release. The digital scale is later also be retrofitted.
Technical specifications:
Mounting arm: rubber jaws, 360 ° freely rotatable
Working height: 122 cm - 183 cm
Clamping range: 1.9 to 4.5 cm
Folding mechanism: 2 fast closing clamps
Foot: HD folding mechanism 6061 T6 tubes
Material: 6061 T6 aluminum
Max. Load: 25 kg
Features: Carrying Case
Dimensions: 117 x 17.8 cm (folded)
Weight: 6 kg
|Product weight:


text:
There are many people who have the desire to open their own business. Most don't know what type of business to start or how to figure that out. Enterprises TV offers advice and suggestions to those who want a piece of the American dream.
One of the first things prospective business owners should consider is where their expertise lies. It makes sense to do work in a field or industry which is already known rather than one which is not. Another factor to consider is what type of business would be best? There are different types such as franchise, E-commerce, proprietor of a brand-new business or an existing one?
- Aspiring business owners should first determine which of the above types fits best into their life. Other considerations might be:
- What are the long-term business goals?
- How much money is there to invest in the business?
- How much time is needed to spend on the business?
- Who can help with web design, E-commerce back-end tools, franchise challenges?
- What are the specific challenges to opening each type of business?
The Enterprises TV show suggests compiling a list of small businesses in the local area. Contact each one and ask if it is okay to meet to talk about specific challenges and successes. This is one of the best methods to determining the right business to open.


text:
After going to Finders Keepers yesterday, I went to the Flemington market today. It was a very market-y weekend.
Lady spinning!!
French caramel man!! I confess some of his caramel made it home with me…
This one has pantaloons!!!
And the dude totally put them on children's HEADS (this is MY PLAN)
Also, cupcake! With tiny, soggy oreo on it!
I confess that, unlike yesterday, I did buy a number of things today. Lots of gorgeous fruit (including quinces), olives, soap.
Today's market was way less crowded than Finders Keepers but that was a good thing in my book. I ate a corn fritter thing. It had a dodgy band playing 90s covers and was much harder to get to that Finders Keepers, but it was a lot more authentic, old-school market-y (one of my companions suggested it had a touch of the Country Women's Association about it — I like that).
But it's way too far away from me to go to regularly. Ah well…


text:
PopGun Presents
Mother Feather
Glasslands Gallery
289 Kent Avenue
Brooklyn, NY, 11249
8:30 PM (event ends at 11:30 PM)
This event is 21 and over
"If you're feeling this you're my next victim," front-woman Ann Courtney intones with steely-eyed confidence, assuring the audience of motherfeathers of their submission to the cult. With their ferocious and enthralling live show, Mother Feather has ignited a wildfire of followers since hitting the New York club scene in 2010.
Epic songs warrant larger-than-life costumes and dance, and a garishly bedazzled Courtney and hype-woman sidekick Elizabeth Carena slither, thrust, and high-kick their way through a set that invokes Marc Bolan's cheeky
swagger, Bowie's braggadocio, and the howl and shimmy of a "To Bring You My Love"-era PJ Harvey. This is pop cock-rock: anthemic riffs, now-or-never bravado, and tight dance beats bolstered by pop precision and purity of mission. "'Mother Feather' is the muse," says Courtney. "It is the embodiment of your wildest inspiration. Every song is a challenge -- to myself and the audience. I want catharsis and I want it now."
Tickets Available at the Door
Upcoming Events
Glasslands Gallery
- Sorry, there are currently no upcoming events.


text:
Re: Nicholas Ault, Lunenberg, axe-maker
In reply to:
Re: Nicholas Ault, Lunenberg, axe-maker
Patsy Ault 4/09/06
I am not an Ault descendant. I am writing a book on Ontario axe-makers and I want the help of the Ault family in including a proper bio of Nicholas Ault, to recognize his place in the trade. I have a broad axe made by him, marked "N. AULT" twice. There is a marriage bond for Cornwall, Apr 18, 1834 for Nicholas Ault of Osnabruck and Nancy Empey of Williamsburg. Nicholas appears in the ledger of A. Hurd & Co of Maitland, as a journeyman axe-maker, about 1835. He is listed as an axe and edge tool manufacturer in Lenenberg in 1865-6. In 1871 he is a 57 year old blacksmith in Osnabruck. In 1881 he is a 67 year old blacksmith, wife Nancy, daughter Mary, in Osnabruck. I have a list of children (no source) Nancy 1842, James 1844, John 1847, Richard 1850, William, 1852, Abraham 1855 and James. Their birthplaces given as Napanee, Osnabruck or Matilda. Is he a descendant of Nicholas of the KRRNY?


text:
Bradley Beach Best Pizza
Vic's, located in Bradley Beach, New Jersey, is known throughout the state for their superb thin crust pizz. Located right next door to the Bradley Beach Cinema, Vic's Pizzeria & Restaurant is a convenient stop when you are going to a show.
Try out the Best Pizza in Bradley Beach, you can't go wrong.


text:
- PriceStarting at $29.95/month
- FrequencyMonthly
- Items per Box7-10
- Shipping Time3-5 days
- Free Shipping?Yes
- Ships toUS
- Date ShippedFirst week of the month
- Average Box Rating 0 out of 5 based on subscriber reviews
Hey Gluten Free
Hey Gluten Free is a monthly subscription service that was created to make your life a lot easier and more delicious. Every month you will receive a box of different goods all gluten-free, soy free, corn free, low in sugar and non-GMO.
We make sure to get the best and most delicious products just for you. Get delicious gluten-free goodies delivered to your doorstep every month.


text:
Building a Network of Young Leaders for the Future of the Philippines
Filipino Family Fund sponsors Youth Leadership Development programs for empowering college students with challenges to make a difference in their communities locally and in areas devastated by Super Typhoon Haiyan. To learn more about human rights, they attend weekly meetings on Capitol Hill. The students also have mentors in non-profit management, agriculture, and ministry. These challenging projects provide excellent foundations for their future careers.


text:
Doug's Photo Blog
An offering of images from Charleston, South Carolina, and travels elsewhere.
Tuesday, June 5, 2012
Bella Italia
Window, near Piazza Navona, Rome, Italy © Doug Hickok All Rights Reserved
(Velvia slide film. Click on image for larger view.)
This Roman vignette offers a glimpse of colors and textures typically seen in beautiful Italy. I am suddenly craving a large plate of lasagna and a glass of wine!
Bella Italia!
Posted by
Doug Hickok
14 comments:
Email This
Share to Twitter
Share to Facebook
Share to Pinterest
Newer Posts
Older Posts
Subscribe to:
Posts (Atom)


text:
Results 1 to 2 of 2
03-28-09, 14:15 #1Registered User
- Join Date
- Mar 2009
- Posts
- 1
Unanswered: Access db2 with other users than db2inst1
Hi there,
installed db2 on Ubuntu 8.10 today using the standard users (e.g. db2inst1). Everything went fine and I could create a database in the control center (db2cc). Now I want to use the database with other users on the same host (!) as well.
I do have a java tool which accesses the database. It works fine with user db2inst1 but not with another standard user of the same host.
Do I need a special configuration to allow other users as well to access the host? I couldn't find it in the documentation.
03-28-09, 18:12 #2Registered User
- Join Date
- May 2003
- Location
- Posts
- 5,737
Did you grant access to that user? See the GRANT statement in the SQL Reference Vol 2.M. A. Feldman
IBM Certified DBA on DB2 for Linux, UNIX, and Windows
IBM Certified DBA on DB2 for z/OS and OS/390


text:
I first wrote the phrase "build with, not for" about a year ago today. It was a parting thought from a longer piece (So You Think You Want to Run a Hackathon? Think Again.) critiquing the-default-to-hackathon impulse that still seems to take hold whenever a government or non-profit or other sort of institution is confronted by the simultaneous desire to do something with tech and to build community.
"Build community."
Over the last decade, spurred by the rise of certain technologies (social media! smartphones! internet!?!?) and the popularity of populism (*~*crowdfunding*~*), "community building" has taken on the air of a finite project, a Thing that organizations in and out of the tech world can report having Done if they host a happy hour, build an email list, slap up a social media account, or run a hackathon.
But we know that communities and their construction are far more complicated than that.
Communities are, and operate through, relationships.
Relationships, social capital: if "community building" is meaningful as a practice, it is as the practice of forging, supporting, and stewarding relationships not just between an organization and a buncha individuals, but between those individuals. (This is one of the biggest differences between a "community" and a traditional "audience" — which will come into play in a later post when we examine common strategic missteps non-profits and governments make.)
Relationships take time. And work. Being their steward and spurer requires less the skill of reading a recipe* and more the skill to…improvise. It's the skill of being present enough with the group of people you're trying to engage that you see the communities they already participate in and find opportunities to work in partnership. It's the skill of listening and stepping back before volunteering to create something new. (Sound familiar?)
It's the skill of participating.
If you don't participate in the lives of those you seek to organize, you can't form relationships to these people and you can't inspire them to form relationships to each other. No relationships? No community. It's that simple.
Over the next little while, I'm going to dig further into these issues, examining relationship and community building as they relate to civic engagement, with a particular eye to challenges and best practices relevant to technology and governance. (Separately and together.) The nuts and bolts of building with, not for.
To kick things off, I want to return to my piece from last year and share with you the reflection that got me started on my own journey with co-design:
In the civic tech space, we talk a lot about "open source" — code made available for anyone's reuse and remixing, with the knowledge that opening up to the contributions of many can allow for the discovery of problems, patterns, new ideas, and opportunities that one alone might miss.
Open format events are like open source social code — rough rubrics that can be "forked" (borrowed and altered) to mobilize people around technology.
But you have to be smart about it.
Dragging and dropping a format for community-building without first evaluating the context in which you're organizing and who you're organizing for is a recipe for exclusion and redundancy. The reason we didn't jump into a hackathon at the Funk Parade was because we knew, after evaluating our criteria for a "good" event in that context and the people we wanted to bring together, that a hackathon wasn't the right fit — but that didn't mean that we couldn't do anything technical. It was just a design challenge, prompting us to ask in real terms (not aspirational) what structures we needed to borrow, what attributes we had to re-imagine (and, worst-comes-to-worst, create)that would allow us to bring together the most diverse group of DC-ers possible.
Diversity is about a lot more than race and gender. It's about age, class, background, profession, sexuality, neighborhood, World Cup team…the many ways in which people identify and are identified. When we willfully ignore diversity in the design of our technology, our social spaces, and our "community meet-ups", we trivialize a future where universal technology access is meaningful and dynamic — not just about literally having access to tools.
So here's my plea for "building community" around civic tech, whether in the context of an event, a new project, or something else all together:
Build with, not for.
Be honest about the in-groups you're dealing with, invest time in thinking about the impact of place (online and off) on who gets to engage and how, and borrow liberally and often from ideas you see working. (Actually working, that is.)
In creating The Tech Embassy, we had the luxury of knowing many of the alternative structures that we could borrow from. But now, you do, too.
Onward and upward, cuties.
// L(e)
*Three hackathons, a dash of happy hour, a pinch of salt, and call me in the morning.


text: Yesterday I upgraded Jewellery by Jaymie to allow for new stock and lots more pieces to be added again this will be a long process more likely then not as want to re-shoot images as the lighting annoys me with the white background as never comes out the same colour background in all photos! But I have added New pieces to Jewellery by Jaymie, first up to be blogged our the new little Animal friends (except Dapper Cat who has made a return to the shop! More styles of him to come) Tea & Bunny necklaces and this little Hedgehog fella. Let me know what you think? And any possible animals you might like to see? Mr Pug necklace will be back up on the shop as well just need to take some new photographs of it.


text:
Discussion in 'Shoes and Apparel' started by ClubHoUno, Jul 21, 2008.
Anyone knows where I can buy this Fila Bjorn Borg jacket:
cool jacket. Yeah i want it too
I just bought in a local shop in Scandinavia - but I hoped to get it for cheaper on a US based tennis shop, but not so
I paid $200 for it - but it's worth it.
It's really high quality - with a combination of wool and acryl (don't know what acryl is called in English). I also will buy the blue and green version of this - it looks great
Will post some pics of it in the "Show your apparel section" later - I still have it in the washer - always wash the new stuff I buy, before I wear it
Um...it's made of wool. I don't know why you would want to put it in the washer. I hope you didn't ruin it. I have this same jacket and I've never washed mine. But then again I almost never wear it because I don't want to have to wash it. :shock:
that is one nice jacket i want it 2
It's 30% wool AND 70% ACRYL (some kind of synthetic fabric) - it will go in the washer just fine. 30 degrees celcius
I'm looking forward to getting the fresh dark blue and the grass green version of this tennis jacket. Definitely not one I would play in, it's just too hot and NOT sweat wicking at all. I prefer my Sergio Tacchini tennis jackets if I need to use a tennis jacket on the court.
This BB Fila jacket is just for show to and from the court and maybe also as a nice sweater for daily use
I bought a size XXL in the Fila BB Jacket - and it only just fits me. The 2 Sergio Tacchini tennis jackets I bought were also XXL - and they are almost too big, so remember that this Fila is tight and small sized - but great looking
Separate names with a comma.


text:
About Us:
Tru-D SmartUVC is the only UV disinfection device backed by a CDC-funded, randomized clinical trial to show that it is capable of reducing the transmission of health care-associated infections by up to 30 percent. More Tru-Ds have been deployed to disinfect hospitals across the globe than all other competing look-alike offerings, fighting deadly pathogens such as C. diff, MRSA, CRE, VRE, MERS, Ebola and many more. An effective and innovative technology backed by sound science, Tru-D SmartUVC is on a mission to eradicate HAIs making hospitals safer places for patients and staff.


text:
From Russian escalation to a negotiated solution in Syria
From Russian escalation to a negotiated solution in Syria
By J. Dana Stuster
November 16, 2015 | THE HILL
Secretary of State John Kerry's meetings with counterparts from the Gulf states, Iran, and Russia in Vienna represent the best chance the United States has for resolving Syria's civil war. While Russia's recent escalation clearly risks prolonging the violence needlessly, it could turn out to be the catalyst for a resolution to the crisis. Russia's strikes so far and the fact that the meetings are happening at all are a reason to pursue this diplomatic opening, especially when the United States has few options in Syria short of risking a direct military conflict with Russia.
When Russia began conducting airstrikes in defense of the Assad regime a month ago, the assessment of many Kremlinologists was that it was the latest manifestation of President Vladimir Putin's impulsive action bias. "The question is, have the Russians thought two or three moves ahead?" Andrew Weiss of the Carnegie Endowment for International Peace said. "This to me looks suspiciously like what happened in Ukraine, where what seemed like a good idea, a very pressurized decision by the Russians to unleash aggression against Ukraine, has backfired quite badly."
Now it is increasingly clear that Russia is not trying to retake all of Syria. The Russian escalation, even supported by the approximately 2,000 Iranian troops the United States believes is operating with Assad regime forces, is not of the scale necessary to completely shift the balance of power in Syria. As Daniel Serwer noted at the outset, even if Russia were to fill the new barracks they've constructed, it would constitute "a far bigger commitment than the few advisors Russia has maintained in Syria in the past, but it is not a force likely to make much of a difference…in the ongoing civil war there." Russia is targeting opposition forces in Aleppo and Idlib, but is not aggressively striking Kurdish militias or the Islamic State.
Outside of Syria, Moscow has been participating in a series of consultations with other foreign leaders about the civil war. There was Assad's late-night visit to the Kremlin, then talks with the United States, Turkey, and Saudi Arabia in Vienna, some coordination with Iran, and even Russian claims that it was meeting with Syrian rebel groups. Russia appears to be looking for a way to wind down the war sooner rather than later.
There is an international consensus that the only sustainable conclusion to the crisis in Syria is a negotiated political solution; this is frequently acknowledged by President Obama. Even Assad's erstwhile arms-supplier China reiterated its support for peace talks at U.N. General Assembly. But there's also a sense of resignation and diplomatic complacency with the stagnated civil war.


text:
What Is Bid Size & Ask Size With Common Stock?by Slav Fedorov ; Updated April 19, 2017
Bid is the highest price at which an investor can sell a stock; ask is the lowest price at which an investor can buy a stock. An extended quote typically provides the highest bid and lowest ask prices, with sizes. The bid size is the number of shares that someone is willing to buy at that price; the ask size is the number of shares that someone is willing to sell at that price.
A quote for XYZ that reads: bid $25.48 1,000, ask $25.55 800 means that a buyer is currently willing to buy 1,000 shares of XYZ at $25.48 and a seller is willing to sell 800 shares of XYZ at $25.55.
Extended Quote Structure
Retail investors usually transact at market prices (the best buy or sell prices they can get), but many stock orders are entered with a specified price at various levels below or above the current market price. Orders always specify the number of shares a trader wants to buy or sell. The highest offer to buy and the lowest offer to sell become the current bid and ask, with size, or the number of shares, specified in the order.
Indications of Interest
Market makers and specialists (professional traders who often take the other side of retail trades, as well as large institutional traders) often provide indications of interest -- how many shares they are willing to buy or sell at a specific price. Indications of interest from professional traders often form the current quote, with its bid and ask.
Order Size and Trading Volume
Bid and ask size are of little importance to retail investors in large-cap stocks that trade millions of shares daily because there are always buyers and sellers ready to transact at the current prices in sufficient quantity. If the current ask size is 1,000 shares and an investor buys 1,000 shares, the ask will go back to 1,000 shares in a blink. However, in low-volume stocks that only trade several thousand shares a day, the current bid and ask sizes are important because that may be all the shares traders are willing to buy or sell at current prices. For example: An investor who wants to sell 1,000 shares of XYZ, which only trades 20,000 shares a day, sees a current bid to buy 500 shares at $25.48. He may be able to sell 500 at that price, after which the next bid down, which could be 300 at $25.15, could become the current market bid.
- "PassTrak Series 7: General Securities Representative License Exam"; Dearborn Financial Services; 2003