text:
On a recent holiday in Cape Town, Eric Chen, a director at Chinese aerial photography company DJI, flew his drone "quadcopter" off the top of Lion's Head for a stunning airborne GoPro-filmed tour of Table Mountain, the city bowl, rocky shores and smooth beaches, seals playing in the surf and great white shark cage-diving.
South Africa punches well above its weight in international luxury travel, according to year's prestigious Condé Nast Traveler Reader's Choice Awards: the country garnered the top spot in the best 100 hotels and resorts, and Cape Town was voted fourth out of the world's 25 must-see cities.
Johannesburg, South Africa's major city and economic capital, has made a dramatic comeback after white flight, overcrowding and neglect led to the decline of the inner city, writes Simon Allison.
The 37-year-old Cape Argus Tour is billed "the world's largest timed cycle race". In its first year, 525 entrants rode the route. Now, it attracts 35 000 ardent cyclists, including big Tour de France names.
The prized rooibos, indigenous to the south-western reaches of South Africa, has long been used by the Khoi and San as a refreshing drink and a medicinal herb. Today it is enjoyed around the world. Its following has grown so vast that a pair of enterprising sisters in a small rural town has set up the Rooibos Route.
Eight days, 800 kilometres and some seriously steep ravines – this is the Absa Cape Epic, one of the most extreme things a person can do with a bicycle. The hors catégorie event is the most televised mountain bike stage race in the world. Each year a new route is chosen, as difficult and as beautiful as the last.
Imagine floating in a natural rock pool right on the lip of the largest waterfall in the world, looking down into the gorge far, far below as the waters of the great Zambezi river thunder past, the spray rises and perpetual rainbows dance in the air.
Sign up now for free high-resolution photos of the rainbow nation.


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Question About Using the Living Social Deal at AJ Madison
I talked to one of the sales people at AJ Madison who said that when you use the Living Social deal, you lose all their sale prices and discounts, so if your appliance has that 10% today offer, you lose that. If it's got the "lower price in cart" discount, you lose that. Memorial Day (and presumably other special sale) prices don't apply.
But I'm pretty sure someone here said they used the LS deal and got the AJ Madison discounts.
So, for those of you who have actually used the deal, what's your experience?
I haven't redeemed mine yet, but I think the $250 savings is better than 10% off unless you are buying something really expensive.
Also, one could easily split their purchase by using the $500 voucher on one transaction, then do a separate purchase for other things they need where they can get a sale or 10% discount.
Remember that unless you are one of those who gets the whole discount for free, you paid $250 for the privilege of buying something at regular price. Don't forget to add that $250 back to the equation. You're really only getting $250 off when it comes down to it. That might be a good deal for a DW or other appliance less than $2500, but for a fridge or stove that's higher priced, it's probably cheaper to just get the sale price.
And the deal is over. When I called today to inquire about my order, not only were they extremely rude to me, they actually hung up on me! :-o The rep also stated that they pulled the LS deal because so many customers were wanting the free shipping that had been advertised for Memorial Day and/or the free shipping that would come standard on their items. Of course people wanted it--AJ Madison advertised it! (FYI I could not get the FS that is always offered on my item- they actually tacked on an extra $99 to the $3,000 range I was purchasing.)
Overall I'm disappointed in their service and will not do business with them in the future.

subset: commoncrawl


text:
Collected Dreamy Tracks
12. Jul. 2011, 8:41 von red-green-blue
iTunes survey
11. Sep. 2006, 0:42 von sweetvenusLast edited: 2 December 2006
How many songs: 2,361
Sort by song
First Song: #1 Crush
Last Song: Zoot Suit Riot
Sort by time:
Shortest Song: Let's Hear It For Nine Inch Nails (0:03)
Longest Song: Phase III (19:24)
Sort by artist:
First Artist: "Weird Al" Yankovic
Last Artist: Zero 7
Sort by album:
First Album: (What's the Story) Morning Glory?
Last Album: Zoot Suit Riot
Top Three Most Played Songs (on desktop):
1. Flake (10 times)
2. I Can't Win (10 times)
2. My Goddess (9 times)
First song that comes up on Shuffle: Be Yourself
"sex", how many songs come up? 21
"death", how many songs come up? 9
"love", how many songs come up? 63
"you", how many songs come up? 170
Number of Songs Starting with "I", "I'm", "I'll", "I've", etc.? 48
Song With the Most Different Versions: Head Like a Hole (6 versions)


text:
Custom Recipes for
Julia's Best Ever Chocolate Chip Cookies In A Jar
Bunny cakes, lamb cakes, chocolate eggs, and carrot cakes to nibble on.
Breakfast casseroles, drinks, fancy eggs, and brunch ideas for a sweet Easter event.
Delicious recipes, party ideas, and cooking tips! Get a year of Allrecipes magazine for $7.99!
See how to make the chocolate lover's chocolate chip cookie.
See how to bake the perfect chocolate chip cookie.
Try these addictive cookies, featuring milk-chocolate chips. Mmm.
Read our allrecipes.com blog


text:
Glamorise Response:
Thank you for your feedback. It sounds like you may be wearing the incorrect size. Also, given that you are 5'3" the garment may feel long on the torso. Check out HerRoom's Bra Fitting Advice Page and Universal Cup Size Page with help on sizing.
Martha (pending 5.7.12) from Ann Arbor, MI.


text:
With less than a 0.1 percent share of Japan's car market, Alfa Romeo knew it couldn't match the marketing muscle of local automakers such as Toyota Motor Corp., which together make nine out of 10 vehicles sold in the country.
Japan may further damage already frayed relations with China if it moves military forces closer to a group of islets claimed by both nations, said Jeff Kingston , director of Asian Studies at Temple University's Tokyo campus.
Prime Minister Naoto Kan 's moves to weaken Japan's currency and shake up his administration have made him more popular as he seeks broader support for his economic agenda after surviving a leadership challenge.
"A landslide victory will put wind in his sails and give him and the LDP two additional years to get on with their reform agenda."
- Jeff Kingston on Dec 03, 2014


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Show off your love for Tokyo with the Turbo Tokyo Brief. This suit is perfect for both swim and water polo and it is complete with a fully lined, durable fabric to let you train longer and harder.
Bought these for my son for high school swim team and water polo practice.He loves them. Double wall construction makes it feel more substantial than the super thin briefs we unknowingly bought in the past. It is low rise which gives him street cred with the polo players. Even though it seems skimpier to Mom and I he feels the thickness gives him more support and therefore modesty and confidence. We used the size chart and it fit perfect. Pick your print but we're very happy with this Turbo water polo suit. Worth every penny.
use it all the time for waterpolo practices and i love the feeling and the fit.
Get an alert when we add new styles of Turbo! Simply sign up and we'll notify you.


text:
New Stem Cell State
December 10, 2014|
Through cellular reprogramming, researchers have produced a novel pluripotent mouse stem cell in vitro.
December 10, 2014|
Through cellular reprogramming, researchers have produced a novel pluripotent mouse stem cell in vitro.
December 9, 2014|
For the first time, researchers have mimicked biological evolution using chemicals instead of living organisms.
December 1, 2014|
By horizontal gene transfer, an antibacterial gene family has dispersed to a plant, an insect, several fungi, and an archaeon.
December 1, 2014|
Researchers are turning to venom peptides to protect crops from their most devastating pests.
December 1, 2014|
In Chapter 7, "Green Evolution, Green Revolution," author John Archibald describes how endosymbiosis helped color the Earth in a verdant hue.
December 1, 2014|
Your Atomic Self, Eureka!, A Talent for Friendship, and Undeniable
December 1, 2014|
Bat populations ravaged; hundreds of amphibian species driven to extinction; diverse groups of birds threatened. Taking risks will be necessary to control deadly wildlife pathogens.
November 27, 2014|
Whole-genome sequences of 16 different mosquito species reveal rapid evolution and could inform malaria research.
November 24, 2014|
The cold-tolerant cereal crop allowed humans to live and farm higher than ever starting more than 3,000 years ago.
November 19, 2014|
Researchers discover a densovirus that is strongly associated with sea star wasting disease.


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Matt Vensel is in his first year at the Star Tribune after covering the Ravens for the Baltimore Sun for six years. He is a Pittsburgh native and a Penn State grad. Follow him at @mattvensel.
Mark Craig has covered the NFL for 23 years, and the Vikings since 2003 for the Star Tribune. He is one of 44 Pro Football Hall of Fame selectors. Follow him at @markcraignfl.
Master Tesfatsion is the Star Tribune's digital Vikings writer. He is a 2013 graduate of Arizona State and worked for mlb.com before arriving in Minneapolis. Follow him at @masterstrib.
Vikings coach Leslie Frazier and his staff received some positive news this afternoon when it was announced they had officially been chosen to coach the North squad in the Senior Bowl later this month. The event annually features the nation's best senior college football standouts and top NFL draft prospects and is considered a premier showcase as NFL teams prepare for April's draft.
The game itself will kickoff at 3 p.m. on Jan. 28 in Mobile, Ala. But a week's worth of events will begin on Jan. 22. In a statement released by the Senior Bowl, Frazier expressed excitement in the news that the Vikings coaches would have a hands-on opportunity to interact with players.
"It gives us the chance to work with some of the most talented football players in the country as they prepare to enter the NFL," Frazier said. "We are looking forward to working with and getting to know all of the players we'll have the opportunity to coach. We are excited to go into a great environment in Mobile and work with some incredible prospects."
Mike Shanahan and his Redskins coaching staff will lead the South squad at the event.
"We're really excited about having both of these staffs here in Mobile working with the future stars of the National Football League," Senior Bowl president and CEO Steve Hale said. "With the amount of experience that both Coach Shanahan and Coach Frazier bring to the table, our players will receive the very best instruction as they prepare for the next level."
In last year's NFL draft, the Vikings selected five players who had participated in the Senior Bowl, including quarterback Christian Ponder.
Frazier last coached at the event in 2004 when he was the defensive coordinator with the Cincinnati Bengals.
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Winnipeg Free Press - PRINT EDITION
Deal with your marriage before making a move
DEAR MISS LONELYHEARTS: I can't control myself any longer. I'm happily married, but my marriage is lacking something. I don't know what it is. I've found myself connecting with, and thinking about, one of our sales reps at work. When I get up I look forward to coming to work and seeing him. He's loud, but always happy. He was hit by a car one day, and still came to work that morning. Since then, I've had a very special place in my heart for him. He's a strong man, who sounds like he was raised by a wonderful mother. He has children and grandchildren he talks about all the time. He's a great guy -- so different from my husband. I want to start something, but we work together and I know how these kind of romances can go so wrong, but I just feel like I need to take a chance.
I don't know if I am ready to leave my husband for him, and I think this guy might have a girlfriend, but I don't want to wonder later "what if?" He talks about hanging out at the Palomino Club, so I am thinking of asking him to meet me there, but not sure if that'd be the right thing to do. Should I approach him and let him know how I feel, and see where things go?I He does give me the impression he might like me too. Or, should I leave my husband first? Should I tell my husband that I am thinking of this other man? Please help! -- Lonely Lady Wanting a Chance at the Title, Winnipeg
Dear Lonely: You admired the sales rep because he dragged himself into work when he could have used the accident to pretend he needed time off. An honourable guy like this does not want a dishonourable woman. Until you decide you want to break up your marriage, you should not ask this guy to meet you at the Palomino Club or anywhere else. You will look cheap and dishonourable. The best you can do is say, "I wish I'd married a guy who is more like you" and see if that conversation goes anywhere. If you and your husband go to counselling and can't work things out, and break up honourably, THEN you can approach this guy after a few months have passed. But, you can tell him you have broken up, right after it happens. His eyes might light up or he might just say, "Best of luck in your new life."
Dear Miss Lonelyhearts: My wife is way too fat. She eats like a glutton when we go out for dinner, which is her favourite pastime. She was in the bathroom with me and stepped on the scale and I looked over her shoulder and saw the number. I said, "YOU WEIGH 205 POUNDS!" She broke down and cried and said "It's your fault!" My fault? She weighted 135 when we got married five years ago. I know it's shallow of me to tell her she has to lose 50 of the 70 pounds or I'm going to say goodbye, but our plans to have children aren't even feasible at 205. She's a walking heart attack at this point. Do I know if I still love her? I don't know. We hardly ever have sex because she's too exhausted after eating dinner. How could she let herself go like that? I am still the same size guy she married. -- So Disgusted, Winnipeg
Dear Disgusted: Why did she gain 70 dangerous pounds? Why is she stuffing herself? Does it have anything to do with an empty marriage? Did it go cold a while back? And how about you? How do you feel about HER as a person, aside from the weight? After five years you've gone from loving her enough to marry her, to not sure you care. Is there a conflict over having a baby? Sometimes people put on a lot of weight for protection against something -- having sex, or getting pregnant or against feeling lonely. Gaining 70 pounds is a huge statement. It's crisis time. Get into counselling together.
Republished from the Winnipeg Free Press print edition November 20, 2013 C4
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Police: Alaska man crashed pickup truck, stole forklift while intoxicated to retrieve it11:37 AM 0
FAIRBANKS, Alaska - Police in Fairbanks, Alaska, say a 21-year-old man drove a stolen forklift more than 3 miles while ...
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text:
After being discarded, Clown makes his way through town having a series of adventures as he tries to find a home for himself and his other toy friends.
Publisher: New York : H. Holt, 1996.
Edition: 1st American ed.
Characteristics: 1 v. (unpaged) :,col. ill. ;,32 cm.
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Christmas MistletoeA traditional kiss under the mistletoe is a well-loved part of the Christmas celebrations not to be missed. Hanging a fresh sprig of hand-tied mistletoe in a prominent place adds a touch of fun and also perpetuates a tradition that dates right back to ancient times.
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Sunnyvale, CA, United States Job Type: Full-Time IP Infusion provides a comprehensive set of Layer 2 and Layer 3 switching and routing solutions for communication equipment vendors. Our scalable, modular technology enables OEMs and ODMs to develop innovative access, enterprise,2/17/2015 Sunnyvale, CA Save This Job
PERFORMANCE PROFILE: Boingo Wireless is looking for a highly energetic and customer focused person with comprehensive knowledge of IP-based network systems, including Ethernet and WiFi. This is a hands-on role requiring technical savvy and experience working independently2/16/2015 Edinburgh, IN Save This Job
Peak 10 provides IT infrastructure solutions that ensure the 24/7/365 availability and security of our customers? critical data and applications. Customer-centric and cost-competitive, Peak 10 IT infrastructure solutions are designed to scale and adapt to customers? changing2/1/2015 Charlotte, NC Save This Job
Join Intellinote! This is a unique opportunity to get in on the ground floor of a rapidly-growing and well-funded software startup with over 10,000 customers in an incredibly hot space. As a lead developer, you?ll work with cutting-edge technologies, be given as much1/31/2015 Reston, VA Save This Job
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Location: Bedford, MA Description: As a member of the SaaS Operations Team at FirstBest, the Production Support Engineer will work closely with your team, R&D Engineers, and our Professional Services team to diagnose and resolve technical issues for your customers. You will1/31/2015 Bedford, MA Save This Job
Xoom Corporation (Nasdaq: XOOM), one of the fastest growing digital money transfer companies in the world is revolutionizing the international money transfer market by providing people with an easy, convenient and cost-effective means of sending money worldwide. We offer a1/31/2015 San Francisco, CA Save This Job
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Do you like to use your technical skills to solve problems for other people Bit9 + Carbon Black, the leader in endpoint threat prevention, detection and response is growing and looking for exceptional engineers to join our team. We need w2/27/2015 Waltham, MA Save This Job
PERFORMANCE PROFILE: The Quality Management team is an integral part of the software delivery process at Boingo Wireless. We are looking for a Sr. QA Engineer that will be responsible for testing Boingo products (Web Apps, Billing, Internal Tools/Products on different devices),3/18/2015 Los Angeles, CA Save This Job
Requisition Number15-0023 Post Date2/17/2015 TitleField Service Engineer (I09) CityMartinsville StateIN DescriptionSummary Provides highly visible customer support through the performance of on-site installation, as well as overseeing any necessary diagnoses, troubleshooting,3/17/2015 Martinsville, IN Save This Job
Location: Pasadena, CA Department: Engineering, Operations Type: Full Time Min. Experience: Experienced As a rapidly growing start-up, Metacloud now apart of Cisco, offers a unique opportunity for talented, passionate, and highly-creative systems engineers to have a wide-ranging3/13/2015 Pasadena, CA Save This Job
Empirix stands alone at the forefront of network performance solutions that preempt technology issues, ensure peak level performance, and predict the smartest actions for delighting customers, controlling costs, and optimizing business processes. Empirix solutions guard against3/11/2015 Bedford, MA Save This Job
About the company: NetBase is a world-class social analytics start-up, chosen by world-famous brands such as Coca Cola, Johnson & Johnson, Ralph Lauren, Yum Foods and Nickelodeon to deliver social, live analytics for action ? through industry leading speed and accuracy. Located3/11/2015 Mountain View, CA Save This Job
Nashville, TN Serves as technical WiFi engineer by exhibiting expertise within the ratified IEEE 802.11 WiFi standards that encompass the ISM band frequencies and technologies that provide wireless mobility across LAN infrastructure. Work closely with ENA?s contractors, field3/4/2015 Nashville, TN Save This Job
Get Smart Content?s (GSC) new Technical Support and Sales Engineer will combine a technical product understanding with sales and customer support skills to deliver support and advice related to our B2B enterprise accounts. The emphasis of the work ranges from handling S3/S43/2/2015 Austin, TX Save This Job
Description: InPhenix has an immediate opening for an Optical Sales Application Engineer with a unique combination of technical competence and professional sales skills. Responsibilities: The position, located and based in our Livermore, CA offices, will promote, sell and2/25/2015 Livermore, CA Save This Job
Job Overview As our Web QA Engineer, you will be responsible for ensuring the best user experience for Scribd on the web. You are a strong generalist who will become an expert on all aspects of the Scribd product, in a fast-paced environment without a lot of formal process. When2/25/2015 San Francisco, CA Save This Job
Why Our Company Catbird brings the excitement of the start-up, a fun and collaborative work environment, with a culture focused on innovation and excellence. The company's products are cutting edge, loved by our customers who see the impact of better security and the direct2/25/2015 Scotts Valley, CA Save This Job


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Free live music
Unwind at Crossroads every weekend. See who's playing.
Fresh additions
Crossroads is bringing you more to see, more to do. Find out what's new.
2nd Saturday Family Night
Music, games, food and fun for the whole family. Learn more.
2nd Saturdays
A family-friendly evening at Crossroads
The second Saturday of every month
Family game time, kids' music performer, free popcorn, balloon performer, and so much more!
This event is free.
Northwest Folklife Presents:
Cultural Arts Series
3rd Saturday of every month
6:30pm at the Market Stage
Once a month Northwest Folklife comes to Crossroads to host an open evening of music, dance, and merriment! Experience different types of folk music, learn a few dance steps, then spending the evening dancing with your family and friends. All ages and abilities are welcome.
Dusty Strings Concert Series
3rd Thursday of every month
6:30pm at the Market Stage
Crossroads is teaming up with Dusty Strings Music School to present the Dusty Strings Concert Series. Free concerts at the Crossroads Market Stage on the fourth Thursday of each month feature performers drawn from Dusty Strings' respected faculty. Every show in this continuing series will be different, featuring the wide range of genres and styles mastered by Dusty Strings' teacher-performers.
Cultural Crossroads Festival
November 7 - 9, 2014
Market Stage & South Concourse
Our annual multi-cultural celebration features performances from 35 ethnic music and dance groups, plus an international bazaar filled with handmade crafts.
Crossroads is Green
Find out what Crossroads is doing to help make the environment healthier for our community.
Family Singalong
April 4 & June 6
11:00am at the Market Stage
Spring Break Art Camp
April 6 - 10
at The Studio Fine Art Classes
Music at the Market Stage
Free live music performances take place at the Market Stage every Thursday, Friday, and Saturday night.
Birthday Parties
Crossroads has so many ways to celebrate your child's birthday!
Farmers Market
Tuesdays from 12:00 - 6:30pm
East Parking Lot
Fresh Washington produce, flowers, music, and connections to organizations that make this a great neighborhood. Thank you for a wonderful 2014 Season. Our 2015 Season begins May 26.
Come See What's New!
We've added new retail stores, restaurants and entertainment for the whole family.


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As Is taken with Eos Digital 400D TSI
Taken at Cleveland Metropark Zoo
Taken at the Cleveland Metroparks Zoo
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Oh, I'd love to snuggle those paws! What a lovely shot.
awww so cute
What a great photo! Beautiful composition.
The leopard looks so peaceful. Beautiful shot!
beautifully taken, i do love those big paws
This is a great shot , but i disagree with animals in zoos , we have a zoo in edinburgh which i have visited and there are a couple of jaguars which i have taken a photo of when i was doing some research into zoos and the living conditions were not good , these are big animals and need space , lets face it if we keep endagered animals in zoos and they dont breed successfully they will still become extict , so i feel wild is wild and thats where they should stay.
please see this interesting link
Any photo with a cat, large or small, gets my attention. This is really a good image of a happy contented beautiful animal.
pboast, it is all about balance. We need big animals in the wild and we need them in captivity so they don't go the way of the Tasmanian tiger. Most zoos in Australia do have large areas for the big animals to roam and to breed.
But, back to you, Jeff. I do love your work. And also I have just spent a bit of time in your Favourites as well. Very good taste. Best wishes, Michelle.
snow leopards are some of the most magnificent felines in the world. I hope they will be more than a photograph in twenty years.
Something NEW…
Hi Jeff
Just started a community site where you will be able to post a CAT image for Free
and add a link ( for Free also) to your redbubble gallery.
And You're welcome to post your e-mail address.
The Site name….'' Pictures of Cats '' Lovers.
I shall post a '' Cat of the Day'' feature on a regular base, which means that your
Cat picture and Your redbubble gallery link will be posted for a minimum of one
week on the main page (and one full day right at the top of the page ). Plus a link
to your image will be posted on the archive section, so people can stay in touch
with your work at any given time.
Also You will be able to post Comments and others will be able to do the same.
You can have a look at the Site ( I'm building it at the moment ) and you'll
See that I did use my body Alfred ( the lama ) to give you an idea since I don't
have a Cat on hand. I really like Cats but I'm not allowed to have one where I live.
But You should get the idea. Just go to http://pictures-of-cats-lovers.blogspot.com
So if You like this idea ( some free advertising for your redbubble gallery ) just
follow up by sending me a Cat photo and your redbubble link and I'll set You up.
Or for more details get in touch with me at firstname.lastname@example.com
or try my phone at 418-257-2091
Roger B. ( rog99 )
Great photo, I just wanna cuddle him!


text:
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Day and night, beginners and families, carvers and snowboarders can get their kicks on the red, blue and black slope s on the Zau[:ber]g Semmering. The perfectly prepared slope s and the view across the Semmering-Rax region turn every day of skiing on the Zau[:ber]g Semmering into a unique experience. Our split park offers snowboarders and free-skiers the opportunity to really go wild on panels built specially for that purpose, while incomparable tobogganing fun is provided by our adventure tobogganing run, 3 km long and lit up at night. The ideal winter activity with magical mountain charm for everybody on the lookout for an experience! >> more
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Fax: +43 2664/8038-34
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Snow report info line:
+43 2664 / 2575


text:
Are you Nimesh Gupta?Claim your profile
Publications (12)36.16 Total impact
[Show abstract] [Hide abstract]
ABSTRACT: Abrin toxin is a plant glycoprotein, which is similar in structure and properties to ricin and is obtained from the seeds of Abrus precatorius (jequirity bean). Abrin is highly toxic, with an estimated human fatal dose of 0.1-1 μg/kg, and has caused death after accidental and intentional poisoning. Abrin is a potent biological toxin warfare agent. There are no chemical antidotes available against the toxin. Neurological symptoms like delirium, hallucinations, reduced consciousness and generalized seizures were reported in human poisoning cases. Death of a patient with symptoms of acute demyelinating encephalopathy with gastrointestinal bleeding due to ingestion of abrin seeds was reported in India. The aim of this study was to examine both dose and time-dependent transcriptional responses induced by abrin in the adult mouse brain. Mice (n=6) were exposed to 1 and 2 LD50 (2.83 and 5.66 μg/kg respectively) dose of abrin by intraperitoneal route and observed over 3 days. A subset of animals (n=3) were sacrificed at 1 and 2 day intervals for microarray and histopathology analysis. None of the 2 LD50 exposed animals survived till 3 days. The histopathological analysis showed the severe damage in brain and the infiltration of inflammatory cells in a dose and time dependent manner. The abrin exposure resulted in the induction of rapid immune and inflammatory response in brain. Clinical biochemistry parameters like lactate dehydrogenase, aspartate aminotransferase, urea and creatinine showed significant increase at 2-day 2 LD50 exposure. The whole genome microarray data revealed the significant regulation of various pathways like MAPK pathway, cytokine-cytokine receptor interaction, calcium signaling pathway, Jak-STAT signaling pathway and natural killer cell mediated toxicity. The comparison of differential gene expression at both the doses showed dose dependent effects of abrin toxicity. The real-time qRT-PCR analysis of selected genes supported the microarray data. This is the first report on host-gene response using whole genome microarray in an animal model after abrin exposure. The data generated provides leads for developing suitable medical counter measures against abrin poisoning.Toxicology 05/2012; 299(1):33-43. DOI:10.1016/j.tox.2012.05.005 · 3.75 Impact Factor
[Show abstract] [Hide abstract]
ABSTRACT: Japanese encephalitis (JE) is one of the leading causes of acute encephalopathy with the highest mortality rate of 30-50%. The purpose of this study was to understand complex biological processes of host response during the progression of the disease. Virus was subcutaneously administered in mice and brain was used for whole genome expression profiling by cDNA microarray. The comparison between viral replication efficiency and disease progression confirms the active role of host response in immunopathology and disease severity. The histopathological analysis confirms the severe damage in the brain in a time dependent manner. Interestingly, the transcription profile reveals significant and differential expression of various pattern recognition receptors, chemotactic genes and the activation of inflammasome. The increased leukocyte infiltration and aggravated CNS inflammation may be the cause of disease severity. This is the first report that provides a detailed picture of the host transcriptional response in a natural route of exposure and opens up new avenues for potential therapeutic and prophylactic strategies against Japanese encephalitis virus.Virology Journal 03/2011; 8:92. DOI:10.1186/1743-422X-8-92 · 2.09 Impact Factor
Article: Alteration of blood brain barrier permeability by T-2 toxin: Role of MMP-9 and inflammatory cytokines.[Show abstract] [Hide abstract]
ABSTRACT: T-2 toxin is a cytotoxic fungal secondary metabolite produced by different species of Fusarium such as F. sporotichioides, F. poae, F. equiseti, F. acuminatum etc. This class of mycotoxins causes a number of pathologies including nervous disorders, cardiovascular alterations, immunodepression and hemostatic derangements. In the present study, mechanism of T-2 toxin induced alteration of blood-brain barrier (BBB) permeability was assessed in terms of oxidative stress, gene expression of MMP-9, MMP-2 and their inhibitors TIMP-1 and TIMP-2, activation of inflammatory cytokines in both brain and peripheral tissue spleen. Gel zymography was used to show the activity of MMP-9 and MMP-2. The percutaneous exposure of 1 LD50 T2 toxin caused a reversible alteration in BBB permeability as observed by extravasation of Evans blue dye. Maximum dye level was observed on day 3 and reduced by day 7. A significant GSH depletion was observed on days 1 and 3. Brain ROS and lipid peroxidation levels increased significantly on 1 and 3 days and decreased by day 7. The SOD levels in brain showed significantly higher activity on 3 days (4-fold) and 7 days (5-fold) of toxin exposure compared to control. A similar trend was observed with catalase enzyme levels. The gene expression analysis of cNOS and iNOS showed varying levels of expression on different time points of post exposure. MMP-9 expression was significantly high on days 3 and 7 in brain with corresponding alteration in TIMP-1. MMP-2 and TIMP-2 showed no effect. Gene expression analysis of the inflammatory cytokines, IL-1α, IL-1β, IL-6 and TNF-α showed elevated levels on day 7 in brain. As spleen plays an important role in inflammatory response we analyzed MMP-9, MMP-2 and inflammatory cytokines in spleen. The MMP-9 was activated on day 7. MMP-2 activity was found to be elevated on 3 and 7 days and TIMP-2 mRNA level increased on 1 and 3 days in spleen. Inflammatory cytokines, IL-1 α, IL-1β, IL-6 and TNF-α showed elevated levels on days 1 and 3 in spleen indicating an early effect in spleen than in brain. In summary, the results of the study showed that the T-2 induced alteration in BBB permeability is mediated through oxidative stress, activation of MMP-9, and proinflammatory cytokines in brain as well as contribution from peripheral tissue spleen.Toxicology 02/2011; 280(1-2):44-52. DOI:10.1016/j.tox.2010.11.006 · 3.75 Impact Factor
Article: Transcriptional regulation and activation of the mitogen-activated protein kinase pathway after Japanese encephalitis virus infection in neuroblastoma cells.[Show abstract] [Hide abstract]
ABSTRACT: Japanese encephalitis virus (JEV), the most frequent and the single most important cause of encephalitis worldwide, has spread throughout most of Asia. For the development of appropriate and effective therapy, there is an immediate requirement to understand the role of host factors in JEV-induced neuropathogenesis. In the present study, we investigated the role of mitogen-activated protein kinases (MAPKs) in JEV infection of mouse neuroblastoma (N2A) cells. The MAPK pathway was studied at the transcriptional level to access the gene expression profile at different time points after JEV infection. The effector MAPK genes were also analyzed for protein expression and activation. Gene expression analysis showed a significant regulation of extracellular signal-regulated kinases (ERK)1, ERK2 and c-Jun N-terminal kinase (JNK)3 genes along with their downstream transcription factors such as Mef2c, c-Jun and Sfn. Experiments with the JNK inhibitor, SP600125, and the ERK inhibitor, PD98059, showed the involvement of JNK in JEV-induced caspase-3 activation and apoptosis, but ERK1/2 had no effect. Overall, our results show the transcriptional regulation of the MAPK pathway and the essential role of JNK in JEV-induced apoptosis in neuroblastoma cells. These findings provide a new insight into the role of the mitogen- and stress-activated kinases in JEV pathogenesis and opens up new avenues of therapeutics.FEMS Immunology & Medical Microbiology 02/2011; 62(1):110-21. DOI:10.1111/j.1574-695X.2011.00792.x · 2.68 Impact Factor
Article: Modulation of ROS/MAPK signaling pathways by okadaic acid leads to cell death via, mitochondrial mediated caspase-dependent mechanism.[Show abstract] [Hide abstract]
ABSTRACT: Okadaic acid (OA) is a specific and potent protein phosphatase inhibitor and tumor promoter. The present study establishes the role of reactive oxygen species (ROS) and mitogen activated protein kinases in cell death induced by okadaic acid. The study showed that okadaic acid is cytotoxic at 10 nM with an IC50 of 100 nM in U-937 cells. The CVDE assay and mitochondrial dehydrogenase assay showed a time dependent cytotoxicity. The phase contrast visualization of the OA treated cells showed the apoptotic morphology and was confirmed with esterase staining for plasma membrane integrity. OA activated caspases-7, 9 and 3, PARP cleavage and induced nuclear damage in a time and dose dependent manner. Compromised mitochondrial membrane potential, release of cytochrome-c and apoptosis inducing factor confirms the involvement of mitochondria. A time dependent decrease in glutathione levels and a dose dependent increase in ROS with maximum at 30 min were observed. ROS scavenger-N-acetyl cysteine, mitochondrial stabilizer-cyclosporin-A, and broad spectrum caspase inhibitor Z-VAD-FMK inhibited the OA induced caspase-3 activation, DNA damage and cell death but caspase-8 inhibitor had no effect. OA activated p38 MAPK and JNK in a time dependent manner, but not ERK½. MAP kinase inhibitors SB203580, SP600125 and PD98059 confirm the role of p38 MAPK and JNK in OA induced caspase-3 activation and cell death. Over all, our results indicate that OA induces cell death by generation of ROS, and activation of p38 MAPK and JNK, and executed through mitochondrial mediated caspase pathway.Apoptosis 11/2010; 16(2):145-61. DOI:10.1007/s10495-010-0554-0 · 3.61 Impact Factor
Article: Expression profile of Japanese encephalitis virus induced neuroinflammation and its implication in disease severity[Show abstract] [Hide abstract]
ABSTRACT: Host immune response particularly through the induction of proinflammatory cytokines and chemokines in Japanese encephalitis virus infection has not been clearly understood in relation with pathogenicity and disease severity. The newly identified host mediators of pathogenesis could be the future target for diagnostic and therapeutics purpose. We investigated the mechanism of JE virus induced pathogenesis in terms of proinflammatory cytokine and chemokine secretion at molecular level in young one-week-old BALB/c mouse after subcutaneous administration of JEV. Histopathology of brain was done to observe the morphological changes after JEV infection and genes relevant to macrophage activation, chemokine secretion, inflammatory cell infiltration, and blood-brain barrier permeability were examined at their gene and protein expression level for various time points after infection. At 6-day post-infection 100% mortality was observed. At 5-day post-infection, there was a robust expression of proinflammatory cytokines and chemokines with increased number of infiltrating inflammatory cells into the brain. Histopathology data confirms the infiltration of leucocytes and there was a marked upregulation in expression of genes relevant to infiltration. The expression pattern of macrophage receptor CLEC5A/DAP-12 signaling has shown the involvement in this robust neuroinflammation. This is the first report that shows the involvement of monocyte and macrophage receptor CLEC5A in severe inflammatory response in JEV infection of brain. This study at gene expression level provides a hypothesis of neuroinflammation, a new lead in development of appropriate therapeutic, and prophylactics against Japanese encephalitis.Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 09/2010; 49(1):4-10. DOI:10.1016/j.jcv.2010.06.009 · 3.12 Impact Factor
Article: Antibodies against refolded recombinant envelope protein (domain III) of Japanese encephalitis virus inhibit the JEV infection to Porcine Stable Kidney cells.[Show abstract] [Hide abstract]
ABSTRACT: Japanese encephalitis virus (JEV) is a mosquito-borne viral zoonosis of public health importance. Global efforts have been made towards development of vaccine for prevention of Japanese encephalitis. The envelope protein of JEV is associated with viral binding to cellular receptors, membrane fusion, and the induction of protective neutralizing antibody response in hosts. Here we report that the antibodies raised against refolded domain III of envelope protein of JEV neutralize the JE virus and inhibit the JEV infection to Porcine Stable Kidney (PS) cells. A reverse transcriptase-PCR amplified gene encoding domain III of JEV envelope protein was cloned into pET28a+ vector and over expressed in E. coli. The recombinant JEV-DIII protein was purified by affinity chromatography under denaturing conditions. The rJEV-DIII was refolded by oxido-redux shuffle and purified to homogeneity by ion-exchange chromatography. Refolded rJEV-DIII was characterized using biochemical and biophysical methods. The polyclonal antibodies were raised against in vitro refolded rJEV-DIII protein in BALB/c mice with Freunds adjuvant. Ninety percent JEV is neutralized when the serum against refolded rJEV-DIII is used at a dilution of 1:80 as against 60.5% neutralization capacity with the same dilution of serum raised against denatured rJEV-DIII. The method of expression and purification of biologically functional rJEV-DIII protein described in this study may help in better understanding the biology of JE virus and the development of better vaccine candidate. Since the expression system uses E. coli as the heterologous host, the process is easy and amenable to inexpensive scale-up.Protein and Peptide Letters 11/2009; 16(11):1334-41. DOI:10.2174/092986609789353709 · 1.74 Impact Factor
Article: Chemokine profiling of Japanese encephalitis virus-infected mouse neuroblastoma cells by microarray and real-time RT-PCR: Implication in neuropathogenesis[Show abstract] [Hide abstract]
ABSTRACT: Japanese encephalitis (JE) is one of the leading causes of acute encephalopathy affecting children and adolescents in the tropics. JE virus (JEV) infection causes prominent neurological sequelae in approximately one-third of the survivors. In humans, the inflammatory response of CNS consequent to JEV induced viral encephalitis is mediated through chemokines released by various cells of CNS. In the present study, the chemokine profiles of mouse neuroblastoma cells (N2A) following JEV infection was analyzed by cDNA microarray followed by real-time RT-PCR. Eighty mRNA transcripts belonging to various functional classes exhibited significant alterations in gene expression. There was considerable induction of genes involved in apoptosis and anti-viral response. Modified levels of several transcripts involved in proinflammatory and anti-inflammatory processes exemplified the balance between opposing forces during JEV pathogenesis. Other genes displaying altered transcription included those associated with host translation, cellular metabolism, cell cycle, signal transduction, transcriptional regulation, protein trafficking, neurotransmitters, neuron maturation, protein modulators, ER stress and cytoskeletal proteins. The infection of neurons results in the synthesis of proinflammatory chemokines, which are early important mediators of leukocyte recruitment to sites of viral infection. Our results clearly suggest the implication of chemokines in neuropathogenesis of JEV infection leading to neurological sequelae. Pro- and anti-inflammatory agents targeted against chemokines such as CXCL10 may provide possible therapeutic modalities that can mitigate the morbidity associated with JEV infection of the CNS.Virus Research 11/2009; 147(1):107-12. DOI:10.1016/j.virusres.2009.10.018 · 2.83 Impact Factor
Article: Bacterially expressed recombinant envelope protein domain III of Japanese encephalitis virus (rJEV-DIII) elicits Th1 type of immune response in BALB/c mice.[Show abstract] [Hide abstract]
ABSTRACT: Japanese encephalitis is a major cause of encephalitis in Asia. Cases occur largely in rural areas of the South and East Asian region resulting in significant morbidity and mortality. Multiple vaccines exist to control Japanese encephalitis, but all suffer from problems. Envelope protein domain III of Japanese encephalitis virus is involved in binding to host receptors and it contains specific epitopes that elicit virus-neutralizing antibodies. Earlier, the protective efficacy of domain III has been evaluated in mice by some researchers, but these studies are lacking in explanation of humoral and cellular immune responses. We have earlier reported cloning, expression, purification and in vitro refolding of Japanese encephalitis virus envelope protein domain III (rJEV-DIII). Ninety percent JEV is neutralized when the serum against refolded rJEV-DIII is used at a dilution of 1:80. In the present study, we have evaluated the immunomodulatory potential of refolded rJEV-DIII protein in BALB/c mice with Freunds complete/incomplete adjuvants. Mice were tested for humoral immune response by ELISA. Cell-mediated immune response was tested by lymphocyte proliferation assay and cytokine profiling. The rJEV-DIII generated high IgG antibody and its isotypes (IgG2a and IgG3) and induced significant expression of INF-gamma and IL-2 cytokines. The rJEV-DIII induced significant lymphoproliferation of splenocytes. In conclusion rJEV-DIII induced Th1 type of immune response which plays an important role in protection for intracellular pathogens.Vaccine 09/2009; 27(49):6905-9. DOI:10.1016/j.vaccine.2009.09.006 · 3.49 Impact Factor
[Show abstract] [Hide abstract]
ABSTRACT: Okadaic acid (OA) is the major component of diarrhetic shell fish poisoning toxins and a potent inhibitor of protein phosphatase 1 and 2A. We investigated the signal transduction pathways involved in OA induced cell death in HeLa cells. OA induced cytotoxicity and apoptosis at IC50 of 100nM. OA treatment resulted in time dependent increase in reactive oxygen species and depleted intracellular glutathione levels. Loss of mitochondrial membrane permeability led to translocation of bax, cytochrome-c and AIF from mitochondria to cytosol. The cells under fluorescence microscope showed typical apoptotic morphology with condensed chromatin, and nuclear fragmentation. We investigated the mitochondrial-mediated caspase cascade. The time dependent activation and cleavage of of bax, caspases-8, 10, 9, 3 and 7 was observed in Western blot analysis. In addition to caspase-dependent pathway AIF mediated caspase-independent pathway was involved in OA mediated cell death. OA also caused time dependent inhibition of protein phosphatase 2A activity and phosphorylation of p38 and p42/44 MAP kinases. Inhibitor studies with Ac-DEVO-CHO and Z-VAD-FMK could not prevent the phosphorylation of p38 and p42/44 MAP kinases. Our experiments with caspase inhibitors Ac-DEVD-CHO, Z-IETD-FMK and Z-VAD-FMK inhibited capsase-3, 8 cleavages but did not prevent OA-induced apoptosis and DNA fragmentation. Similarly, pretreatment with cyclosporin-A and N-acetylcysteine could not prevent the DNA fragmentation. In summary, the results of our study show that OA induces multiple signal transduction pathways acting either independently or simultaneously leading to apoptosis.Toxicology 12/2008; 256(1-2):118-27. DOI:10.1016/j.tox.2008.11.013 · 3.75 Impact Factor
Article: Immunogenicity of a recombinant envelope domain III protein of dengue virus type-4 with various adjuvants in mice[Show abstract] [Hide abstract]
ABSTRACT: Dengue fever, a mosquito borne viral disease, has become a major public health problem with dramatic expansion in recent decades. Several dengue vaccines are at developing stage, none are yet available for humans. There is no vaccine or antiviral therapy available for dengue fever till date. Domain III of envelope protein is involved in binding to host receptors and it contains type and subtype-specific epitopes that elicit virus neutralizing antibodies. Hence domain III is an attractive vaccine candidate. In the present study we report the immunomodulatory potential of refolded D4EIII protein in combination with various adjuvants (Freunds Complete adjuvant, Montanide ISA720, Alum). Mice were tested for humoral immune responses by ELISA, immunofluorescence assay and plaque reduction neutralization test. Cell mediated immune response was tested by lymphocyte proliferation assay and cytokine profiling. All the formulations resulted in high antibody titers that neutralized the virus entry in vitro. D4EIII in combination with montanide ISA720 and Feuds complete adjuvant gave highest antibody endpoint titers followed by alum. The level of antigen-stimulated splenocyte proliferation and cytokine production was comparable to that obtained from Con A stimulation and cytokine profiling of stimulated splenocyte culture supernatants indicated that all the adjuvant formulations have induced cell mediated immune response as well. These findings suggest that D4EIII in combination with compatible adjuvants is highly immunogenic and can elicit high titer neutralizing antibodies and cell mediated immune response which plays an important role in intracellular infections, which proves that refolded D4EIII can be a potential vaccine candidate.Vaccine 08/2008; 26(36):4655-63. DOI:10.1016/j.vaccine.2008.07.006 · 3.49 Impact Factor
Article: Production of IgM specific recombinant dengue multiepitope protein for early diagnosis of dengue infection.[Show abstract] [Hide abstract]
ABSTRACT: Dengue virus infections have recently undergone dramatic expansion in range, affecting several tropical and subtropical regions of the world. Early detection of dengue infection based on the identification of antibodies has emerged as a practical and reliable means of diagnosis of dengue fever. The recombinant dengue multiepitope (rDME-M) protein specific to IgM in E. coli was produced in a 5-L fermentor for use in diagnostic purpose. After fermentation, dry cell weight was approximately 11.8 g/L of the culture. The rDME-M protein was purified under denaturing conditions using single-step nickel nitrilotriacetate (Ni-NTA) affinity chromatography. The final yield of purified rDME-M protein from this method was approximately 68.5 mg/L of the culture. The purity of rDME-M protein was checked by SDS-PAGE analysis, and the reactivity of this protein was further checked by Western blotting and enzyme-linked immunosorbent assay (ELISA). The purified protein was used as an antigen in the development of an in-house dipstick ELISA and evaluated with a panel of 80 patient sera, characterized using commercially available tests for detection of dengue antibody. The results were in excellent agreement with those of IgM capture ELISA (Pan-Bio) and rapid immunochromatography (IC) test (Pan-Bio). These results show that the in-house dipstick ELISA using rDME-M protein can be used as a promising kit because of its comparable sensitivity, specificity, field applicability, and low cost.Biotechnology Progress 04/2007; 23(2):488-93. DOI:10.1021/bp0602698 · 1.88 Impact Factor
Top Journals
- Toxicology (2)
- Vaccine (1)
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- FEMS Immunology & Medical Microbiology (1)


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I have a 2008 Honda Civic and want to rear face as long as possible. Any suggestions on a seat that will fit my car well? I would like ot keep him in the center seat, so I will be using the seatbelt instead of LATCH. My car only has LATCH on the two side back seats, not the center seat.
2008 civic - best ERF seat?
Comments (1)
Posted 11/24/2012
Graco myride 65
~ Crissy! GO, BabyGear
"Live so that when your children think of fairness and integrity, they think of you." - H. Jackson Brown


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Alexandria company helps fight deadly Ebola virus
Carrying a deadly virus?
Infrared camera could help others know who's ill
A company with an Alexandria connection is part of a global effort to limit the spread of deadly viruses like Ebola.
Gary Strahan of Alexandria is CEO of Infrared Cameras Inc., which developed a thermal imaging system used in places such as airports to spot someone with a fever — a potentially infected person — before that person is allowed to spread an infection globally.
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