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House overlooking I-670 sold, clearing way for 2 more
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- Brian R. Ball
- Staff reporter- Columbus Business First
- Email | Google+ | Other | LinkedIn
That tall, skinny house overlooking Interstate 670 east of the High Street cap has been sold, with expectations two more homes could get built next door this year.
Short North developer Connie Klema's Kerr Street Place Ltd. affiliate sold the three-level, 1,600-square-foot home at Kerr and East Russell streets Jan. 22 for $375,000, not far from the $385,000 she sought.
Klema tells me buyer John Galvin had expected to close in December but a delay in the financing pushed the transfer off a few weeks. Undeterred, Klema said she allowed Galvin to haul his stuff into the house in late December.
"It was in my best interest," she said, "so I took a chance."
Klema and equity partner Al Coughlin Jr. of the Coughlin Automotive Group expect to squeeze two other larger homes on the 0.1-acre sliver of land she bought more than a decade ago. The original vision for the property called for ground-level commercial space and three full-floor condos above. The credit crunch that began in the mid-2000s forced her to shelve those plans. Klema said the second and third homes on the wedge-shaped ground could begin in the spring.
But right now, she said she will concentrate on the redevelopment of an old masonry block warehouse at 52 Brickel St. into two side-by-side condos around the corner from the Kerr Street Place project.
"This is my side job," the real estate attorney reminded me, "so I'm taking my time."
Brian R. Ball covers commercial real estate and development for Columbus Business First.
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Analysis: Can Schmidt beat the boom/bust cycle?
Can Joe Schmidt's Ireland build on their current rich vein of form as the Rugby World Cup looms into view asks Brendan Cole.
Can Joe Schmidt's Ireland build on their current rich vein of form as the Rugby World Cup looms into view asks Brendan Cole.
Pat Lambie has retained his place at flyhalf for South Africa's final northern hemisphere test against Wales in Cardiff on Saturday as coach Heyneke Meyer made eight changes to his match-day squad.
Connacht Rugby and the IRFU have announced that winger Niyi Adeolokun has signed a new two-year contract which will see him remain at the province until at least June 2017.
Ireland flanker Chris Henry has had exploratory heart surgery and will be out of rugby for the foreseeable future, due to what was described as "a defect in his heart wall."
Australia back Kurtley Beale's $45,000 Australian (€30,800) fine for his involvement in a text message scandal will be used to fund a programme for young indigenous Australian women.
After a 2014 that saw Ireland win nine from ten Test matches, outhalf Johnny Sexton speaks about the season that was and what lies ahead for this team in 2015.
Ulster's Paddy Jackson has been ruled out of action for up to six weeks after picking up a shoulder injury in his side's Guinness PRO12 clash with Ospreys on Friday evening.
Damien O'Meara and Wes Liddy are joined by former Irish international Denis Leamy to look back over Ireland's thrilling win over Australia on Saturday.
Ireland coach Joe Schmidt is recovering well after having his appendix removed in St Vincent's Hospital last night.
Treviso missed a late penalty as Leinster hung on to claim a 24-24 Pro12 draw in Italy.
Brian O'Driscoll is pulling out all the stops to mentor Robbie Henshaw through his bid to claim Ireland's vacant 13 shirt.
Lansdowne kept pace with Ulster Bank League Division 1A leaders Terenure College thanks to a bonus point 28-21 victory over Young Munster on the Aviva Stadium's back pitch today.
Michael Cheika has challenged Australia to "harden up" against England at Twickenham on Saturday to dispel any notion the Wallabies are a soft touch.
Joe Schmidt's "ruthless" selection policy has forced Ireland to shed mental weaknesses that were costing victories, according to front-rower Rory Best.
Jamie Heaslip was delighted that Ireland managed to hold on to record a 26-23 victory over Australia in Dublin.
Ireland fended off a relentless late onslaught to hold on for a thrilling 26-23 autumn international win over Australia in Dublin this evening.
Leinster are the the last of the Irish provinces in Pro12 action this weekend, with an afternoon battle away to Benetton Treviso. Tadhg Peavoy previews the action.
England finally got that winning feeling with a 28-9 victory over Samoa at Twickeham following five straight defeats for Stuart Lancaster's side.
Argentina kicked four drop goals, including three by outhalf Nicolas Sanchez, to upset France 18-13 and close their European tour with two wins in three Tests on Saturday.
Ireland coach Joe Schmidt has been taken to hospital suffering from appendicitis - it is understood he sought treatment after Ireland's victory over Australia.


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Digging: Pink Turns Blue - If Two Worlds Kiss
awesome album aside from a couple tracks. aint no From One as expected but still pretty great
Expecting a pizzamachine review. This has some solid tracks (i.e. Brutiful, Through The Valley, Crawling To The Sky etc.).
Hoping this is good. I wasn't impressed with... Super Cali-fragi-listic-expi-ali-dubstep, or whatever it was called.
"Expecting a pizzamachine review."
Yeah I'm defs gonna try writing one
"AOTY". Yeah not even close lol, and this is coming from a huge Ra fan....
• BREAKING: Chad Kroeger Sucks Dick• Refused aren't fuckin dead• System of a Down to embark on Eur..• Chris and Shawn quit Megadeth• Brand New in studio• Katy Perry to headline Super B• The Black Dahlia Murder streams c..• Fall Out Boy set release date for..• Intervals split with vocalist• Callejon debut first material fro..
• Starbomb set to release second LP• Adimiron stream new album• Just Before Dawn stream new LP• New track from Enter Shikari• New Archgoat Song• Third Angels and Airwaves Song• While She Sleeps release track• Engel Debut New Single, Album• Madred stream debut album• CityCop stream new EP• Mekaal Hasan Band reveals live DVD• Primordial post new video• The Decemberists release new video• Owen streaming covers LP• 11Paranoias stream new LP» see all news
Bands: A B C D E F G H I J K L M N O P Q R S T U V W X Y Z


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UPDATED: Medfly eradicated from Mexico
11/04/2014 2:44:10 PMMediterranean fruit fly has been eradicated from Mexico. Ceratitis capitata, a type of fruit fly that... Read more.
Florida citrus season brings higher quality fruit
11/04/2014 6:31:50 PMDUNDEE, Fla. — Compared to last year, this fall is bringing higher quality of Florida citrus... Watch Video.
Diet affects bee sensitivity to pesticides, says study
11/03/2014 1:19:15 PMFeeding honeybees a diet of natural pollen makes them significantly more resistant to pesticides than... Read more.
Florida, North Carolina partner on organic strawberry work
11/03/2014 12:44:52 PMThe University of Florida and North Carolina A&T Universities have received a $200,000 from the... Read more.
Team Produce turns out for City of Hope
11/03/2014 12:30:18 PMDUARTE, Calif. — For the eighth year in a row, Team Produce turned out in force to show support... Watch Video.
Pack Expo poised to break attendance record
11/03/2014 11:39:40 AMCHICAGO — Just hours into Pack Expo 2014, event organizers said they already had almost as many... Watch Video.
UC Davis students design olive oil sensor
10/31/2014 5:36:49 PMIs it real or is it Memorex? So goes the commercial about a brand of cassette tape, but the same... Read more.
Sustainability exchange showcases Ventura County operations
10/31/2014 12:02:26 PMIn conjunction with the recent Produce Marketing Association's Fresh Summit in Anaheim, a group of... Read more.
- Ag labor not part of Obama's immigration plan
- Research looks at dung beetles as a natural clean-up crew
- Web seminar discusses post-harvest phosphorus for spuds
- Dry weather pares Mississippi sweet potato crop
- USDA releases five HLB-tolerant citrus rootstocks
- Nominate a leader for a potato industry award


text:
Don't Miss
- 8 Ayurvedic Recipes for Vegetarians at Thanksgiving
- How Ayurveda Inspires Mindful Eating
- Ayurvedic Nutrition: The Ancient Recipe of Happiness
- The Loving Embrace of Snehana for the Fall
- 4 Everyday Recipes to Strengthen Your Kitchen's Ayurvedic Power
- Understanding Gluten and How to Digest It
- Three Pranayamas and their Ayurvedic / Psychological Effects
- Seasonal Bloom: Vata
- Your Ayurvedic Kitchen: Everyday Basics
- Drop the Utensils and Eat with Your Hands!
All posts by Peggy Breeze
Please Pass the Turmeric and Pepper!
Yes, you read that right. Should turmeric replace your salt at the table? If you aren't quite ready to go that far, how about adding turmeric to the big two spices we all have as staples, salt...
- Posted September 17, 2014
Keeping Spices Spicy
According to Ayurveda, every meal should contain all 6 tastes: Sweet, salty, sour, pungent, bitter and astringent. One of the best ways to accomplish this is through the use of spices. Spices are expensive, so learning how...
- Posted June 23, 2014


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|By PR Newswire
Article Rating:
|March 7, 2014 08:20 PM EST
ROSEVILLE, Calif., March 7, 2014 /PRNewswire/ -- Following the federal Ninth Circuit Court of Appeals' recent affirmations of the fundamental, individual Second Amendment right to carry handguns in public for self-defense, Roseville-based gun rights group The Calguns Foundation has published new resources to assist law-abiding Californians with the process of applying for a license to carry.
"One important element of our Carry License Compliance Initiative is to educate people on the laws and process," said Executive Director Brandon Combs. "The information barrier has been a huge problem in the past, but we're confident that our new tools will help thousands of Californians exercise their Second Amendment rights."
In addition to expanded informational offerings, the Foundation has created a new California Carry License (LTC/CCW) Issue Reporting System, a service that applicants can use to report problems with a local law enforcement agency's application policy or practice.
"By creating a database of the personal experiences of carry license applicants from across the state, we will gain a comprehensive understanding of what's actually happening out there," explained Combs. "These reports will help us determine which licensing authorities to focus our Compliance Initiative resources on, and, when necessary, take legal action against."
Combs noted that the Foundation released its new carry license projections in February, which showed the number of Californians licensed to carry handguns skyrocketing during the first year of effective "shall-issue" regulations.
Using information pulled from the California Department of Justice and the issuance statistics of other "shall-issue" states across the nation, The Calguns Foundation estimated that the number of California carry licensees will grow over 1000%, from about 56,000 today to as many as 1,000,000 in 2015.
"There's no question that the public demand is there," said Combs. "The right to carry is the biggest, most culturally-significant issue in gun rights today. We're going to do whatever it takes to make sure that all law-abiding people are free to exercise their Second Amendment civil rights in California."
Those interested in applying for a California carry license can access The Calguns Foundation's FAQs, download state-standard application forms, review legal information, and report unconstitutional or unlawful policies through its website at http://www.calgunsfoundation.org/carry.
The Calguns Foundation (www.calgunsfoundation.org) is a 501(c)3 non-profit organization that serves its members, supporters, and the public through educational, cultural, and judicial efforts to defend and advance Second Amendment and related civil rights.
Logo - http://photos.prnewswire.com/prnh/20130412/SF94281LOGO
SOURCE The Calguns Foundation
The 3rd International Internet of @ThingsExpo, co-located with the 16th International Cloud Expo - to be held June 9-11, 2015, at the Javits Center in New York City, NY - announces that its Call for Papers is now open. The Internet of Things (IoT) is the biggest idea since the creation of the Worldwide Web more than 20 years ago.
Nov. 28, 2014 05:00 PM EST Reads: 1,266
"We help companies that are using a lot of Software as a Service. We help companies manage and gain visibility into what people are using inside the company and decide to secure them or use standards to lock down or to embrace the adoption of SaaS inside the company," explained Scott Kriz, Co-founder and CEO of Bitium, in this SYS-CON.tv interview at 15th Cloud Expo, held Nov 4–6, 2014, at the Santa Clara Convention Center in Santa Clara, CA.
Nov. 27, 2014 07:00 PM EST Reads: 1,427
15th Cloud Expo, which took place Nov. 4-6, 2014, at the Santa Clara Convention Center in Santa Clara, CA, expanded the conference content of @ThingsExpo, Big Data Expo, and DevOps Summit to include two developer events. IBM held a Bluemix Developer Playground on November 5 and ElasticBox held a Hackathon on November 6. Both events took place on the expo floor. The Bluemix Developer Playground, for developers of all levels, highlighted the ease of use of Bluemix, its services and functionalit...
Nov. 27, 2014 07:00 PM EST Reads: 1,393
The 4th International DevOps Summit, co-located with16th International Cloud Expo – being held June 9-11, 2015, at the Javits Center in New York City, NY – announces that its Call for Papers is now open. Born out of proven success in agile development, cloud computing, and process automation, DevOps is a macro trend you cannot afford to miss. From showcase success stories from early adopters and web-scale businesses, DevOps is expanding to organizations of all sizes, including the world's large...
Nov. 27, 2014 06:00 PM EST Reads: 1,808
Some developers believe that monitoring is a function of the operations team. Some operations teams firmly believe that monitoring the systems they maintain is sufficient to run the business successfully. Most of them are wrong. The complexity of today's applications have gone far and beyond the capabilities of "traditional" system-level monitoring tools and approaches and requires much broader knowledge of business and applications as a whole. The goal of DevOps is to connect all aspects of app...
Nov. 27, 2014 06:00 PM EST Reads: 1,210
SAP is delivering break-through innovation combined with fantastic user experience powered by the market-leading in-memory technology, SAP HANA. In his General Session at 15th Cloud Expo, Thorsten Leiduck, VP ISVs & Digital Commerce, SAP, discussed how SAP and partners provide cloud and hybrid cloud solutions as well as real-time Big Data offerings that help companies of all sizes and industries run better. SAP launched an application challenge to award the most innovative SAP HANA and SAP HANA...
Nov. 27, 2014 05:00 PM EST Reads: 1,378
When an enterprise builds a hybrid IaaS cloud connecting its data center to one or more public clouds, security is often a major topic along with the other challenges involved. Security is closely intertwined with the networking choices made for the hybrid cloud. Traditional networking approaches for building a hybrid cloud try to kludge together the enterprise infrastructure with the public cloud. Consequently this approach requires risky, deep "surgery" including changes to firewalls, subnets...
Nov. 27, 2014 04:45 PM EST Reads: 1,277
Want to enable self-service provisioning of application environments in minutes that mirror production? Can you automatically provide rich data with code-level detail back to the developers when issues occur in production? In his session at DevOps Summit, David Tesar, Microsoft Technical Evangelist on Microsoft Azure and DevOps, will discuss how to accomplish this and more utilizing technologies such as Microsoft Azure, Visual Studio online, and Application Insights in this demo-heavy session.
Nov. 27, 2014 04:45 PM EST Reads: 1,217
DevOps is all about agility. However, you don't want to be on a high-speed bus to nowhere. The right DevOps approach controls velocity with a tight feedback loop that not only consists of operational data but also incorporates business context. With a business context in the decision making, the right business priorities are incorporated, which results in a higher value creation. In his session at DevOps Summit, Todd Rader, Solutions Architect at AppDynamics, discussed key monitoring techniques...
Nov. 27, 2014 04:30 PM EST Reads: 1,280
Cultural, regulatory, environmental, political and economic (CREPE) conditions over the past decade are creating cross-industry solution spaces that require processes and technologies from both the Internet of Things (IoT), and Data Management and Analytics (DMA). These solution spaces are evolving into Sensor Analytics Ecosystems (SAE) that represent significant new opportunities for organizations of all types. Public Utilities throughout the world, providing electricity, natural gas and water,...
Nov. 27, 2014 04:00 PM EST Reads: 1,345
The security devil is always in the details of the attack: the ones you've endured, the ones you prepare yourself to fend off, and the ones that, you fear, will catch you completely unaware and defenseless. The Internet of Things (IoT) is nothing if not an endless proliferation of details. It's the vision of a world in which continuous Internet connectivity and addressability is embedded into a growing range of human artifacts, into the natural world, and even into our smartphones, appliances, a...
Nov. 27, 2014 04:00 PM EST Reads: 1,680
How do APIs and IoT relate? The answer is not as simple as merely adding an API on top of a dumb device, but rather about understanding the architectural patterns for implementing an IoT fabric. There are typically two or three trends: Exposing the device to a management framework Exposing that management framework to a business centric logic Exposing that business layer and data to end users. This last trend is the IoT stack, which involves a new shift in the separation of what stuff happe...
Nov. 27, 2014 03:00 PM EST Reads: 1,355
The Internet of Things is tied together with a thin strand that is known as time. Coincidentally, at the core of nearly all data analytics is a timestamp. When working with time series data there are a few core principles that everyone should consider, especially across datasets where time is the common boundary. In his session at Internet of @ThingsExpo, Jim Scott, Director of Enterprise Strategy & Architecture at MapR Technologies, discussed single-value, geo-spatial, and log time series dat...
Nov. 27, 2014 03:00 PM EST Reads: 1,504
An entirely new security model is needed for the Internet of Things, or is it? Can we save some old and tested controls for this new and different environment? In his session at @ThingsExpo, New York's at the Javits Center, Davi Ottenheimer, EMC Senior Director of Trust, reviewed hands-on lessons with IoT devices and reveal a new risk balance you might not expect. Davi Ottenheimer, EMC Senior Director of Trust, has more than nineteen years' experience managing global security operations and asse...
Nov. 27, 2014 01:00 PM EST Reads: 1,699
The Internet of Things will greatly expand the opportunities for data collection and new business models driven off of that data. In her session at @ThingsExpo, Esmeralda Swartz, CMO of MetraTech, discussed how for this to be effective you not only need to have infrastructure and operational models capable of utilizing this new phenomenon, but increasingly service providers will need to convince a skeptical public to participate. Get ready to show them the money!
Nov. 27, 2014 11:00 AM EST Reads: 1,317


text:
Two veteran Boyd-Buchanan teams showed why they among the
best in the area in A-AA tennis, while Brainerd girls and Central boys also
posted wins Tuesday in high school matches at the Brainerd Courts.
Boyd-Buc vs. CSAS – The defending district champion BBS
girls opened the season with a 9-0 win against visiting Arts and Sciences (0-2),
while the Buccaneer boys stopped CSAS, 7-2 in District 5 A play.
In No.1 singles matches, Visaal Sivakumar (CSAS) defeated
Joe Fairchild, 8-4, while Katie Delay (BBS) stopped Sneha Mahesh, 8-0.
"I am pleased with the way we played and a good start to
the season with a district win," said Boyd-Buchanan coach Mikki Brown.
Next Matches: Thurs. 3-13 … BBS at Signal Mtn … Tues.
3-18 BBS vs. Collegedale (at Brainerd – district match)
Fri. Mar. 14 – Grace Academy vs. Arts and Sciences at
Warner Park
Brainerd vs. Chattanooga Central:
Brainerd Girls 7,
Central 2 … Central Boys 6, Brainerd 3:
It was a good start for the newest program in the district as the
Brainerd girls won, 7-2, and the boys, had a good showing in a 6-3 loss in District
Marquee Match: In one of the best matches of the day,
Brainerd's Brittany Canty edged Alex Bonner, 9-7, at No.1 singles.
Bonner trailed 6-2 before winning five of the next six
games to pull even at 7-7. Canty then won the final two games for the victory to
secure the fifth team point and the team win.
Brainerd also captured the No.1 singles match in boys as
Bernard King III defeated Brett Kranske, but Central's depth came through as
the Purple Pounders won at Nos.2-5 in singles, plus at No. 2 doubles.
Two Central Wins: Kylan Harrison was a double winner for
Central with wins at No. 3 singles and No. 2 doubles with Kranske.
"I am pleased with the effort by our Brainerd teams today,
pleased with good sportsmanship shown by players from both schools and with Boyd-Buchanan
and CSAS also playing here it offered a good high school tennis atmosphere for the fans and players,"
said Brainerd coach Alphonso Canty.
Next Matches: Thurs. Mar. 13 – East Ridge at Brainerd
(district match); Red Bank vs. Central (Lakeside)
Boyd-Buchanan 9, CSAS 0
Katie DeLay (BBS) def. Sneha Mahesh, 8-0
Makenna Thomas (BBS) def. Katie Foster, 8-0
Paige Zombron (BBS) def. Allycia Lee, 8-1
Christina Amos (BBS) def. Shailey Shah, 8-1
Shelby Brown (BBS) def. Kendall Camp, 8-0
Libby Reynolds (BBS) def. Cadence Camp (8-1
DeLay-Thomas (BBS) def. Mahesh-Candence Camp, 8-0
Zombron-Amos (BBS) def. Lauren Emmett-Foster, 8-3
Brown-Reynolds (BBS) def. Kendall Camp-Sterchi, 8-1
Records: BBS (1-0, 1-0), CSAS (0-2 0-1)
Boyd-Buchanan 7, CSAS 2
Visaal Sivakumar (CSAS) def. Joe Fairchild, 8-4
Jake Harden (BBS) def. Chandler Custer, 8-5
Grayson Hartness (BBS) def. Kevin Lee, 8-0
Charlie Driver (BBS) def. Sean Kennon, 8-2
David Zombron (BBS) def. Tristan Greer, 8-0
Luke Hendrix (BB) def. Rupert Gongmon, 8-5
Custer-Sivakumar (CSAS) def. Harden-Hartness, 8-3
Fairchild-Driver (BBS) def. Lee-Congmon, 8-2
David Zombron-Tanner Zombron (BBS) def. Kennon-Greer, 8-4
Records: BBS (1-0, 1-0), CSAS (0-2, 0-1)
Chattanooga Central Boys 6, Brainerd 3
Bernard King III (Br) def. Brett Kranske, 8-2
Riley Hicks (Central) def. Ralph Foster, 8-3
Kylan Harrison (Central) def. Kobeai Ford, 8-1
Phillip Bonnington (Central) def. Timothy Moore, 8-2
Aaron Garrison (Central) def. Cantaus Davis, 8-1
Corey Little (Central ) forfeit win
King-Foster (Br) def. Hicks-Bonnington, 8-5
Kranske-Harrison (Central) def. Ford-Moore, 8-0
Davis-Collins (Br.) def. Little-Garrison, 8-5
Records: Central (1-0, 1-0), Brainerd (0-1, 0-1)
Brainerd Girls 7, Chattanooga Central 2
Brittany Canty (Br) def. Alex Bonner, 9-7
Brianna Canty (Br) def. Emily Lyles, 8-0
Michelle Hoans (Central) def. Kayla Corner, 8-4
Endiga Rice (Central) def. Coreka Reed, 8-4
Dezrael Hutchinson (Br.) def. Alexya Barnes, 8-5
Chyna Parks (Br.) def. Caithryn Clark, 8-6
Brittany Canty-Brianna Canty (Br) def. Lyles-Bonner, 8-1
Corner-Reed (Br) def. Hoang-Barnes, 8-6
Reed-Parks (Br) def. Clark-Rice, 9-7
Records: Brainerd (1-0, 1-0), Central (0-1, 0-1)
contact B.B. Branton at email@example.com
November 27, 2014
Before the season started some people across the country called Chattanooga women's basketball coach Jim Foster crazy for scheduling three top 10 teams – Notre Dame (national runner-up) Stanford ... (click for more)
Jim Foster has won more of his share of big games in his 37 years coaching women's college basketball, first at Vanderbilt and then at Ohio State. He can now say he's done the same at Chattanooga ... (click for more)
A couple of former state champions from Baylor knocked off the defending overall champions in the 24 th Annual Turkey Trot 8K at the Sports Barn East on Thanksgiving Thursday morning. ... (click for more)
Before the season started some people across the country called Chattanooga women's basketball coach Jim Foster crazy for scheduling three top 10 teams – Notre Dame (national runner-up) Stanford and Tennessee - before Christmas. After Wednesday night's UTC shocking 67-63 home win against No.4 Tennessee before 4,160, those same folks might believe coach Foster is crazy like ... (click for more)
Jim Foster has won more of his share of big games in his 37 years coaching women's college basketball, first at Vanderbilt and then at Ohio State. He can now say he's done the same at Chattanooga after his Lady Mocs handed the Tennessee Lady Vols their first loss of the season Wednesday night at McKenzie Arena in front of 4,160 bi-partisan fans. It may have been UTC's ... (click for more)
The man rescued by firefighters from Wednesday's fatal fire at 220 Houser St. remains in critical condition today in the burn unit at the Vanderbilt University Medical Center . He has been identified as Randall Lockhart, 41. His wife, Candy, succumbed to her severe injuries a few hours after she was pulled out of the burning house. The couple's three children escaped ... (click for more)
A woman was assaulted and a man shot in a home invasion on Chamberlain Avenue early Friday morning. At approximately 4:43 a.m., the Chattanooga Police Department responded to 2708 North Chamberlain Ave. for a home invasion/shooting. Officers discovered that a black male forced entry into the residence and assaulted the first victim,Tequla Nicole Watkins, 28. ... (click for more)
As a child I was under the impression that a ghastly, grotesque creature had taken up residence in my bedroom closet, the proverbial "Boogie Man" I suppose. This is not an uncommon thing among children and most likely to some fault of my own as I subjected myself to some pretty iconic 1980's horror flicks. There were other kids in the neighborhood that not only claimed to ... (click for more)
On this, "Black Friday," when frantic shopping and crowded aisles cause some nerves to wear thin, please allow me to share a poem a friend sent me not long ago called "Shoes in Church." Perhaps it will be fitting: SHOES IN CHURCH I showered and shaved. I adjusted my tie. I got there and sat In a pew just in time. Bowing my head in prayer, as I closed my eyes. I saw ... (click for more)


text: "It Will Rain" Fan Made Music Video Posted on 10/17/2011 by Mel · Leave a comment This is such an awesome fan made video of Bruno Mars' song "It Will Rain" from the Breaking Dawn Part 1 Soundtrack! Source / Via I wonder what part of the movie this song will actually be in. Do you have any ideas? Comment below Share this:Share on TumblrPrint Popular Posts & PagesJennifer Lawrence Mentions Kristen in Marie Claire (June issue) ***SPOILER ALERT*** Camp X-Ray HOT SEXY Robsten Video! Patricia McKenzie Talks Rob & Filming the Cosmopolis Sex Scene Kristen & Rob at Hollywood Film Awards - Nov 13


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Stunning portraits of Siamese fighting fish by Visarute Angkatavanich
Bangkok-based photographer Visarute Angkatavanich (previously) continues to capture some of the most elegant portraits of fish we've seen. His intimate, crystal-clear photos of Siamese fighting fish (betta) make it seem as though they are suspended in air instead of water. Angkatavanich recently told that he only started photographing the fish after encountering them for the...
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Once Upon A Dream, photography by Jvdas Berra
"Lips, red as blood. Hair, dark as night. Bring me your heart, my dear dear…" Photo: Jvdas Berra Styling & Art Direction: Federico Laboreau, Maximilian Pizzi @ Flatelier Models: Alexandra Agro, Robin Laird, Amanda Shae, Taylor Rae @ Paragon Model Management, Nahuel Mendoza @ GH Management M&H: Angel Moncada, Gio Lozano, Benito Felix, Bruno Danyel...
Horse.Play, sculpture by Wang Ruilin
As the portrayal of my life status, Horse.Play will continue all the time and present different expressions with variations of positions and situations. Meanwhile, the production of works will promote the change of environments and moods, both of which affect each other. Wang Ruilin: website / behance
Postcards for Ants: A 365-day miniature painting by Lorraine Loots
"365 Postcards for Ants is the second phase of a project started on 1 January 2013, which involved me creating a miniature painting every single day for the entire year. In celebration of our city's designation as World Design Capital 2014, I've decided to do it all over again, and this time all the paintings...
Fly in the blue, project by Simona Smrckova
FLY IN THE BLUE MODEL, MUA BY COCO HAIRSTYLING BY JIŘÍ KOČÍ STYLING BY SIMS ASISTANCE: KAMIL SALIBA Simona Smrckova: website / behance
The magic of fall captured by Alex Greenshpun
"When creating a photograph, my goal is not only to freeze a moment in time, but to capture its most subtle and delicate essence. A connection between two people, a child's laughter, a deep gaze in the eyes of a weary man, the playful light between the leaves of a tree, ripples in a pond,...
Anita Anti, photography
Anita Anti: website / deviantArt / flickr / 500px / facebook
Moving time, art direction by Gem Fletcher
Photographer: Mads Perch Art Director: Gem Fletcher Gem Fletcher: behance
Digital art by Boško Šabaz Šabazović
Boško Šabaz Šabazović: facebook
R2-D2 Empire style, sculpture by Alain Bellino
Sculpture made from antique bronze ornaments. Photos taken in Villa Massena and Parc Chambrun in Nice / France. Alain Bellino was born in Nice in 1955. In the 80's, he discovered the world of metal and ornamentation in his father's workshop where he learnt gold and silver plating and bronze restoration. After many years of...


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Destination resorts at play in Florida Senate gambling plan
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- Dara Kam, The News Service of Florida
Two stand-alone casinos in South Florida and allowing track operators to quit racing horses and dogs while keeping lucrative card rooms and slot machines are among the proposals on the table as the Florida Senate prepares to roll out its gambling overhaul.
Senate Gaming Committee Chairman Garrett Richter said Monday he plans to release a committee bill as early as next week that will address a swath of gambling issues, including a "gaming control" commission, "inconsistencies" in state law and "decoupling" racing from pari-mutuel permits that allow for slot machines and card rooms.
Richter expressed support for one "destination resort" permit each in Broward and Miami-Dade counties. Companies would have to bid for the permits, Richter said.
"There's not a chance that this legislature will consider a bill that will provide for unlimited casinos statewide,'' Richter, R-Naples, said.
But, he added, "I happen to think that a destination resort in Miami would be a good thing for the state of Florida. I think it will attract new revenue dollars.''
House Speaker Will Weatherford, R-Wesley Chapel, told reporters last week that his chamber would not consider an expansion of gambling unless it is accompanied by a constitutional amendment asking voters if they want to have a say in future gambling expansions.
Weatherford proposes setting the bar for gambling activities at whatever is currently allowed and whatever passes during this legislative session. Anything beyond that would require statewide voter approval.
But passage of a committee bill, which Richter said could be 300 pages, remains dubious because of all of the competing interests in the gambling arena.
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Chile striker Mauricio Pinilla gets tattoo of agonising last-minute shot that hit that bar against Brazil
It was the strike that came so close to knocking out hosts
Charles Reynolds
Charles Reynolds is a freelance writer and journalist specialising in sport and the arts. He writes the Premier League previews for The Independent online and writes regularly for The Sport Review.
Tuesday 01 July 2014
You'd imagine it would be a moment that Mauricio Pinilla would never want to re-live, his 119th minute shot cannoning off the crossbar instead of nestling into the net inches below.
LIVE: The latest from Argentina vs Switzerland and Belgium vs United States
However instead of trying to forget the fact that he came so close to scoring the last-minute winner that would have knocked out Brazil and sent an entire nation into despair, the Chile striker has had it immortalised forever in a tattoo on his back.
Pinilla's new ink shows a stylised version of the moment his shot smashed into Julio Cesar's crossbar, with the words 'One centimetre from glory' written below in English.
Pinilla's tattoo
Not that he was ever likely to forget the moment he came so close to ending Brazil's World Cup hoodoo over Chile, but Pinilla now has a permanent reminder to take with him everywhere.
Ultimately Chile lost the game in a penalty shoot-out, with Pinilla seeing his effort saved, although that was seemingly one moment that he decided not to get tattooed on himself.
Video: Chile players return home from World Cup
Despite Chile's loss, thousands of fans still lined the streets of Santiago to welcome the players home after an overall impressive World Cup campaign, with President Michelle Bachelet honouring the returning footballers.
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Kendrys Morales a fit for the Royals?
Jan 3, 2014, 11:25 PM EST
Jim Duquette of MLB Network Radio suggests free agent 1B/DH Kendrys Morales would be a good fit for the Royals. The market for Morales is fairly quiet right now as he is tied to draft pick compensation and doesn't offer much aside from his ability to hit.
However, Duquette also suggests that signing Morales would give the Royals the flexibility to trade current DH Billy Butler for starting pitching. The Blue Jays were interested in acquiring Butler last month and could still be a potential suitor if such a situation were to come up.
Morales has been fairly consistent between 2012 and 2013, posting a .787 and .785 OPS, respectively. No one questions his ability to hit, but his value is dragged down by having to be a DH full-time, or otherwise having to play defense at first base poorly. If the Royals were to sign Morales, they would relinquish their first-round pick in the 2014 draft, the 18th overall pick.
- dinofrank60 - Jan 3, 2014 at 11:30 PM
Somehow, baseball finds a way to crimp free agentmovement, again.
- NYTolstoy - Jan 3, 2014 at 11:32 PM
I don't see much upside for losing the pick. They might as well keep Buttler. Unless they get a major league ready starter there's no sense in signing this guy.
- proudlycanadian - Jan 4, 2014 at 7:52 AM
This story illustrates why Duquette was a failure as a MLB General Manager. It is a lousy idea that he floated and should not to be taken seriously.
- tigers182 - Jan 3, 2014 at 11:45 PM
I'd rather draft the best hitter available at #18, put him right in the majors, and pay him significant less. You'll likely get a better player.
- paperlions - Jan 4, 2014 at 10:26 AM
Number of hitters in the 2014 draft capable of hitting MLB pitching in 2014 or 2015: 0
- doctorofsmuganomics - Jan 3, 2014 at 11:59 PM
Keep the draft pick, and stop trying to trade Billy Butler.
- unclemosesgreen - Jan 5, 2014 at 9:49 AM
I've never understood the lack of respect for Billy Butler. Professional hitter. You could do worse than to have both he and Morales, but if you're getting Morales and giving up a draft pick and then paying him a lot more than Butler … seems like trading a dollar bill for two quarters.
Thank you, Doctor, I feel better. Where do I send the check?
- jwbiii - Jan 4, 2014 at 12:13 AM
A DH with 1/$9m left on his contract (or 2/$12.5m) isn't going to get much of a starting pitcher.
- clydeserra - Jan 4, 2014 at 11:32 AM
Houston seems like a better landing spot. I know they have Wallace and Carter though
- unclemosesgreen - Jan 5, 2014 at 9:50 AM
I agree that Houston seems like a better landing spot, but I think it's *because* they have Wallace and Carter.
- kcrobert10 - Jan 4, 2014 at 11:48 AM
Not sure the fascination with trading butler. Yes he had a down yr. He is still 26 and still led the team in rbi and on base %. Ya can't use a guy like that.
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I'm excited to find out the results of the UPCAT. When do you think it would be released? Oh, Happy Holidays! :3
An even better Alyssa Valdez, the struggling Lady Bulldogs and UST's wonderkid. Find out other big impressions in week 1 of UAAP Volleyball here!read more
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Paul Lee nailed a crucial three-pointer plus two pressure packed free throws to lead his team to victory over the NLEX Road Warriors, 95-93.read more
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I'm excited to find out the results of the UPCAT. When do you think it would be released? Oh, Happy Holidays! :3
Last edited by icewind21; Dec 27, 2012 at 04:09 PM.
Hi icewind21! I have a PM. Please clear some space in your inbox.
Alright. My inbox is now cleared. You can send your pm now^


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The 16 best sports quotes of all time
The best from Mike Tyson, Muhammad Ali, Charles Barkley, John McEnroe and more.
The best from Mike Tyson, Muhammad Ali, Charles Barkley, John McEnroe and more.
Upon the 40th anniversary of "The Rumble in the Jungle," Muhammad Ali's third wife, Veronica Porche, told USA TODAY Sports (…)
There was big news in the worlds of technology and social status yesterday. Apple unveiled the iPhone 6 and the iPhone 6 plus. Both are (…)
Ali is The Rock's favorite athlete.
These photos of Muhammad Ali, curated from more than 3,000 images, are now public for the first time from USA TODAY Sports Images. (…)
Kevin Durant has been on a historic scoring tear lately and appears destined to eventually retire as one of the greatest basketball (…)
JERSEY CITY – What kind of impact would a young Muhammad Ali have had in the Twitter era? If you believe Seahawks defensive end (…)


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The making of the Circles of Influence mural painted at Makerhaus in Seattle, Washington by Scott Fuller.
Partnership, friendship and community. A simple hand shake represents the partnership and friendships that are created from working in a collaborative environment. The infinity handshake symbolizes a community and the pattern repeat suggests a larger network outside your own circle of influence.
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text: Hogadon Extends Ski Season By Rodeo Rick March 27, 2013 3:46 PM Share on Facebook Share on Twitter Hogadon Ski Area announces that they will remain open for an extra three days after the scheduled closing date of March 31st, 2013. The area will be open April 5th, April 6th, and April 7th from 8am-4pm. Hogadon will not be open during the week prior to the season extension (April 3rd- April 5th). The Hogadon summer office hours will go into effect on April 8th. Summer hours will be 8am-4pm Monday through Thursday. For more information regarding Hogadon's extended season dates, please call 307-235-8499 or log onto www.hogadon.net. Filed Under: Hogadon ski area Category: Casper News, Sports Share on Facebook Share on Twitter More Leave a comment Send to a friend! Print this page Pin it! Reddit This! Share on Tumblr! More News from K2 Radio New Public Restrooms Open In Downtown Casper Department of Health Develops a Plan for Medicaid Expansion Here Desmond Triplett Accused Of Sex Crimes Education Governance Study Now Available on Legislature's Website Best of the Web


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Role of cell surface GM3 ganglioside and sialic acid in the antitumor activity of a GM3-based vaccine in the murine B16 melanoma model.
ABSTRACT To examine the role of GM3 monosialoganglioside and sialic acid in the antitumor activity of a vaccine based on GM3, hydrophobically conjugated with the outer-membrane-protein complex from Neisseria meningitidis (GM3/VSSP).
In order to evaluate the relationship between antitumor activity and the presence of GM3 on the surface of tumor cells, we used two murine tumor cell lines with different ganglioside expression. Syngeneic mice were immunized with four i.m. doses of GM3/VSSP (120 micro g) at 14-day intervals and challenged subcutaneously with tumor cells.
B16 melanoma cells showed GM3 on cell surface and GM3-dependent in vitro growth. As expected, preimmunization with the vaccine significantly inhibited tumor formation and prolonged survival in mice challenged with B16 cells. In contrast, no antitumor effect was observed in mice challenged with GM3-negative F3II mammary carcinoma cells. The reactivity of sera from immunized mice against B16 cells was confirmed by flow cytometry and immunoperoxidase staining. Depletion of sialic acid residues from the cell surface completely abolished antibody response against melanoma cells.
These results indicate that the antitumor activity of GM3/VSSP is associated with GM3 expression on tumor cell surface and demonstrate a major role of sialic acid in the humoral response of vaccinated mice.
Article: A conjugate of an oligosaccharide fragment of tumor-associated ganglioside antigen with hemocyanin is a prototype antitumor vaccine.[Show abstract] [Hide abstract]
ABSTRACT: Specific stimulation of the immune system against tumor antigens, which may result in elimination of malignant cells, may be performed with the use of vaccines containing a specific native or synthetic tumorassociated antigen. To develop a prototype antitumor vaccine, we synthesized an oligosaccharide fragment of tumor-associated antigen, obtained its conjugate with the protein hemocyanin, and studied the immunogenicity of this conjugate. We developed a technique of immunization of experimental animals by the conjugate and obtained secondary immune response to the tumor antigen with a predominant generation of IgG antibodies (ABs). We showed that the ABs generated after immunization of animals with the use of conjugate reacted with the native tumor-associated antigen. The serum of animals immunized with the conjugate contained IgG1 and IgG3 ABs necessary for specific interaction of macrophages and natural killers with malignant cells; hence, the conjugate may be used as a prototype antitumor vaccine.Doklady Biological Sciences 07/2007; 415:298-301.
Article: Sialic acid and GM3 ganglioside expression in papillomavirus-associated urinary bladder tumours of cattle with chronic enzootic haematuria.[Show abstract] [Hide abstract]
ABSTRACT: This study was based on 30 papillomavirus-associated urinary bladder tumours from cattle with chronic haematuria, the animals having been kept since birth on pasture rich in bracken fern. The ganglioside content was assessed and compared with that of normal bovine urinary bladders, which was shown to be 28.6+/-3.3 (mean+/-SD) microg of lipid-bound sialic acid per gram of fresh tissue. In neoplastic bladder samples this value was higher but variable (120.9+/-80.6 in benign tumours, and 94.7+/-45.7 in malignant tumours). The main ganglioside, GM3, represented ca 75% of the total ganglioside mixture in normal tissues and 50-80% in tumour samples. GM1, GM2, GD1a, GD3 and FucGM1 were found as minor components. The study suggested that GM3 ganglioside may have a crucial role in "downregulation" of the metastatic potential of bovine urothelial cancers.Journal of Comparative Pathology 01/2007; 137(2-3):87-93. · 1.38 Impact Factor
- Carbohydrate-Based Vaccines and Immunotherapies, 10/2008: pages 227 - 262; , ISBN: 9780470473283
Page 1
Mariano R. Gabri Æ Giselle V. Ripoll Æ Daniel F. Alonso
Daniel E. Go ´ mez
Role of cell surface GM3 ganglioside and sialic acid in the antitumor
activity of a GM3-based vaccine in the murine B16 melanoma model
Received: 3 June 2002 /Accepted: 3 September 2002 / Published online: 14 November 2002
? Springer-Verlag 2002
Abstract Purpose: To examine the role of GM3 mon-
osialoganglioside and sialic acid in the antitumor activ-
ity of a vaccine based on GM3, hydrophobically
conjugated with the outer-membrane-protein complex
from Neisseria meningitidis (GM3/VSSP). Methods: In
order to evaluate the relationship between antitumor
activity and the presence of GM3 on the surface of tu-
mor cells, we used two murine tumor cell lines with
different ganglioside expression. Syngeneic mice were
immunized with four i.m. doses of GM3/VSSP (120 lg)
at 14-day intervals and challenged subcutaneously with
tumor cells. Results: B16 melanoma cells showed GM3
on cell surface and GM3-dependent in vitro growth. As
expected, preimmunization with the vaccine significantly
inhibited tumor formation and prolonged survival in
mice challenged with B16 cells. In contrast, no antitu-
mor effect was observed in mice challenged with GM3-
negative F3II mammary carcinoma cells. The reactivity
of sera from immunized mice against B16 cells was
confirmed by flow cytometry and immunoperoxidase
staining. Depletion of sialic acid residues from the cell
surface completely abolished antibody response against
melanoma cells. Conclusions: These results indicate that
the antitumor activity of GM3/VSSP is associated with
GM3 expression on tumor cell surface and demonstrate
a major role of sialic acid in the humoral response of
vaccinated mice.
Keywords Immunotherapy Æ Sialic acid Æ GM3 Æ
Melanoma Æ Breast cancer
Gangliosides are sialic acid-containing glycosphingoli-
pids that have increased expression in the outer leaflet of
cell membranes in various malignancies, including mel-
anoma and breast cancer. Previous works have demon-
strated that gangliosides play a role in cell signaling,
adhesion, and migration of tumor cells (Iwabuchi et al.
1998a). Certain gangliosides, such as GD3, GM2, and
GM3, are typified as tumor-associated antigens, which
make them a good target for cancer immunotherapy
(Chapman et al. 2000; Ragupathi et al. 2000). Anti-
ganglioside-specific humoral immune response has been
obtained in several preclinical or clinical protocols of
active immunotherapy, and titers correlated with mice
or patient survival (Zhang et al. 1998; Ritter et al. 1995;
Helling et al. 1994; Livingston et al. 1994; Ravindranath
et al. 1994a).
In 1999, Estevez et al. described a novel cancer vac-
cine based onthe monosialoganglioside
hydrophobically conjugated with the outer-membrane-
protein complex (OMP) from Neisseria meningitidis to
form very small-sized proteoliposomes (GM3/VSSP)
(Estevez et al. 1999). It is known that GM3 is poorly
immunogenic when administered as the unmodified
nominal antigen (Livingston et al. 1989). Recently, we
have demonstrated that ganglioside presentation in a
proteolipid environment using GM3/VSSP vaccine can
induce a strong antibody response and a potent antitu-
mor effect against B16 mouse melanoma, a syngeneic
tumor model expressing high amounts of GM3 (Carr
et al. 2001; Alonso et al.. 1999). Correlation between
antitumor activity of the vaccine and the presence of
substantial amounts of GM3 on cancer cell surfaces,
however, has not been yet explored using other animal
Antibodies are considered the primary mechanism for
elimination of pathogens, as well as circulating tumor
cells and micrometastases (Zhang et al. 1998). Further-
more, anti-ganglioside antibodies may be effective to
J Cancer Res Clin Oncol (2002) 128: 669–677
DOI 10.1007/s00432-002-0385-7
M.R. Gabri (&) Æ G.V. Ripoll Æ D.F. Alonso Æ D.E. Go ´ mez
Laboratory of Molecular Oncology,
Department of Science and Technology,
Quilmes National University, Bernal B1876BXD,
Buenos Aires, Argentina
E-mail: address: email@example.com
Tel.: +54-11-43657100
Fax: +54-11-43657132
Page 2
induce complement-mediated lysis of malignant cells
(Parajuli et al. 2001).
The great majority of cell surface antigens recog-
nized by antibodies are carbohydrates. In this regard,
carbohydrates have proven to be potent targets for
immune recognition and attack, because of their im-
munogenicity and abundance at the cell surface (Liv-
ingston et al. 1997). Several monoclonal antibodies
have been produced in mice immunized either with
(Chapman et al. 1990; Cheung et al. 1985; Hirabashi
et al. 1985). However, limited information is available
concerning the specific reactive epitope of these mon-
oclonal antibodies. In addition, there is incomplete ev-
idence about the critical reactive residues recognized by
antisera in the surface of tumor cells using ganglioside-
based vaccines.
In the present work, we have examined the antigenic
role of sialic acid in the antitumor activity of GM3/
VSSP vaccine against GM3-positive B16 melanoma
cells. Removal of sialic acid residues from tumor cell
surface completely abolished the reactivity of serum
from vaccinated mice against B16 cells. Interestingly, no
significant antitumor effect was observed in mice
preimmunized with GM3/VSSP and challenged with
GM3-negative F3II mammary carcinoma cells.
Materials and methods
Tumor cells and culture conditions
The murine tumor cell lines B16 melanoma and F3II mammary
carcinoma, syngeneic for C57BL/6 and BALB/c mice, respectively,
were employed. We decided to use the parental B16 cell line (sub-
line F0), that is characterized by the predominant presence of the
monosialoganglioside GM3 on the cell surface (Nores et al. 1987;
Hirabayashi et al. 1985). B16 melanoma cells were maintained in
Dulbecco's modified Eagle medium (DMEM) from Gibco BRL
(Grand Island, N.Y., USA), containing 10% heat-inactivated fetal
bovine serum (FBS), 2 mM glutamine, 10 lg/ml tetracyclin, 100 U/
ml penicillin, 100 lg/ml streptomycin, and 0.25 lg/ml amphoteri-
cin B. The sarcomatoid mammary carcinoma cell line F3II is a
highly invasive and metastatic variant positive for oligosialogan-
gliosides (Vazquez et al. 2000; Alonso et al. 1996). F3II mammary
carcinoma cells were maintained in minimal essential medium
(MEM) from Gibco, supplemented with 5% FBS, 2 mM gluta-
mine, and 80 lg/ml gentamycin.
Cell growth
The effect of exogenous GM3 on the growth of B16 and F3II cells
was assessed in vitro. GM3 ganglioside was obtained from Sigma
Chemical (St. Louis, Mo., USA). Tumor cells were plated onto 96-
well plates (5–10·103cells/well) in medium supplemented with
FBS. The next day, tumor cells were extensively washed with
phosphate-buffered saline, pH 7.4 (PBS) and incubated for 48 h in
the same medium with or without 5% FBS, and in the presence of
indicated concentrations of GM3 ganglioside (0–100 lg/ml).
Monolayers were then washed, fixed, and stained with toluidine
blue. The amount of tumor cells was estimated from absorbance
readings at 595 nm after solubilization with 1% SDS, as reported
(Alonso et al. 1998).
C57BL/6 and BALB/c inbred mice were purchased from the Ani-
mal Care Division, Angel H. Roffo Institute of Oncology (Buenos
Aires, Argentina), and kept 5–10 mice/cage with water and food ad
libitum. Animals with an age of 8–14 weeks and an average weight
of 25 g were used.
GM3/VSSP vaccine was produced and generously provided by the
Center of Molecular Immunology (Habana, Cuba). Briefly, mon-
osialoganglioside GM3 was purified from canine red blood cells
and hydrophobically conjugated with the carrier OMP from N.
meningitidis strain 385, as described in detail (Estevez et al. 1999).
The method allowed gangliosides and proteins to incorporate into
very small size proteoliposomes and conferred high solubility to the
conjugate. Each GM3/VSSP vaccine contained 120 lg in 0.05 ml
of PBS, mixed with an equal volume of the immunological adju-
vant Montanide ISA 51 (SEPPIC, France) prior to injection.
Vaccines were administered i.m. in the quadriceps four times at
14-day intervals. Control animals received only the adjuvant mixed
with PBS.
Mouse sera and monoclonal antibody
Mice were immunized as described above, and 7–21 days after the
fourth dose of the vaccine were bled through the optical artery and
immediately sacrificed. Normal sera were also obtained from un-
treated mice. Sera were frozen at –20 ?C in aliquots for further
analysis. The GM3 ganglioside-specific mouse IgM monoclonal
antibody (clone M2590) was purchased from International Bio-
science (Tokyo, Japan).
Flow cytometry
Cultured B16 or F3II cells were harvested with a cell-scraper,
resuspended in serum-free medium, and then incubated at 5·105
to 1·106cells per sample with 1 lg of the monoclonal antibody
M2590 or a control monoclonal antibody of the same isotype
(IgM) for 30 min at 4 ?C. Tumor cells were washed in medium
containing 1% FBS and further incubated with 5 ll of FITC-
conjugated goat antibodies anti-mouse IgM (Caltag Laboratories,
Burlingame, Calif., USA). Alternatively, tumor cells were incu-
bated with 15 ll of sera of either treated or control mice and then
labeled with FITC-conjugated goat antibodies anti-mouse im-
munoglobulins (Caltag). The cells were then analyzed with a
Cytoron flow cytometer (Ortho Diagnostic Systems, Raritan,
Enzyme-linked immunosorbent assay (ELISA)
Flexible 96-well microtiter plates (Becton Dickinson, Oxnard, Ca-
lif., USA) were coated using 10 lg/ml of the GM3/VSSP conjugate
in carbonate buffer, pH 9.6. Coated and control uncoated plates
were blocked with 2% bovine serum albumin (BSA) in carbonate
buffer and 5-fold serial dilutions of sera were applied. Bound IgM
or IgG antibodies were detected with sequential incubations with
either biotinylated anti-l chain or anti-c chain antibodies (Jackson
Immunochemicals), followed by alkaline phosphatase-conjugated
Extravidin (Sigma) and the chromogenic substrate 4-nitrophenyl-
phosphate (Boehringer Mannheim). All steps were separated by six
washes with 0.1% Tween-20 in PBS. Immunologicals were diluted
in 0.2% BSA in PBS.
Specific antibodies against GM3 ganglioside were detected ba-
sically as described by Ravindranath et al. (Ravindranath et al.
1994b). Briefly, PolySorp 96-well microtiter plates (Nunc) were
coated with 200 ng/well of unconjugated GM3 (4 lg/ml in etha-
nol). Ethanol was added to control wells. Once dry, plates were
Page 3
blocked without previous washing with 2% BSA in PBS. Further
steps were carried out as described above, except that 0.1% BSA/
0.1% Tween-20 in PBS was used for washing.
Lymphoproliferation assay
Splenocytes were resuspended in DMEM supplemented with 10%
FBS and 2 mM glutamine, and plated at 2·105cells per well in U-
bottom 96-well microtiter plates. GM3/VSSP was added at 5 lg
ganglioside/ml. The plates were then incubated at 37 ?C under 5%
CO2for 3 days. Tritiated thymidine was added at 1 lCi/well and
the plates were further incubated for another 18 h. Cells were then
harvested, and tritium incorporation was monitored in a liquid
scintillation counter. The stimulation index was defined as the ratio
of mean cpm from wells supplemented with GM3/VSSP to mean
cpm from wells with culture medium alone (triplicate determina-
Indirect immunoperoxidase staining and neuraminidase treatment
For detection of B16- and F3II-reactive serum antibodies, the
Vectastain ABC kit (Vector Laboratories, Burlingame, Calif.,
USA) was employed on formalin-fixed tumor cell monolayers and
tumor sections following the instructions of the manufacturer.
Monolayers were fixed with formalin and preserved at 4 ?C.
Tumor tissue specimens were fixed with formalin, included in
paraffin, cut in 5-lm sections and rehydrated. Monolayers and
tumor sections were incubated for 1 h with pooled sera at a di-
lution of 1:500 in PBS. Specimens were then incubated with
biotinylated horse anti-mouse immunoglobulins for 30 min, fol-
30 min. Finally, bound antibodies were detected by incubation
with the peroxidase substrate diaminobenzidine for 2–4 min and
counterstained with hematoxylin. Pertinent specificity tests were
performed, including blocking of endogenous peroxidase, omis-
sion of the first serum and utilization of diluted serum from
control mice.
In order to evaluate the role of sialic acid in serum antibody
response of GM3/VSSP vaccinated mice, fixed tumor cells were
exposed to neuraminidase, a hydrolase that removes sialic acid
residues from the cell surface. Tumor cell monolayers or tumor
sections were incubated at 37 ?C for 4 h or overnight, respectively,
in the presence 0.1–1 UI/ml of type II neuraminidase (Sigma)
previous to immunoperoxidase staining. As neuraminidase prepa-
rations may contain protease traces, the presence of proteolytic
activity was discarded by a radial caseinolytic assay using casein-
agarose substrates (Alonso et al. 1998).
avidin-biotincomplex for
Tumor cell challenge
For injection into mice, tumor cell suspensions were prepared by
detaching the cells from culture flasks by trypsinization, followed
by washing and resuspension in culture medium. Tumor cells
were counted and viability was assayed by the trypan blue ex-
clusion test. Twenty-one days after the fourth immunization with
GM3/VSSP vaccine, mice were given inoculations in the subcutis
of the right flank with 0.2 ml of tumor cell suspensions in serum-
free medium. Preliminary experiments showed that a challenge of
2.5·103B16 melanoma cells in C57BL/6 mice and 5·104F3II
mammary carcinoma cells in BALB/c mice induced a similar
tumor incidence, of at least 50%. The respective syngeneic mice
received fourfold the described tumor burdens, in order to
achieve a tumor incidence of 100% (1·104B16 cells and 2·105
F3II cells). In all cases, the time of appearance of local tumors
was monitored by palpation and further confirmed by histopa-
thology. Tumor size was measured with a caliper twice a week
and tumor volume was calculated by the formula: p/6·width2·
length. When tumors exceeded 15,000 mm3, animals were killed
and necropsied.
Expression of gangliosides in B16 melanoma and F3II
mammary carcinoma cells and effect of exogenous GM3
on in vitro growth
To assess the presence or absence of GM3 ganglioside in
the surface of B16 and F3II tumor cells, flow cytometric
detection using the specific anti-GM3 monoclonal anti-
body M2590 was performed. As shown in Fig. 1a, the
antibody reacted brightly against B16 melanoma cells
and the reaction was notably inhibited by preincubation
with soluble GM3. On the contrary (Fig. 1b), no inter-
action with the surface of F3II mammary carcinoma
cells could be observed. These results confirm a signifi-
cant expression of GM3 in B16 cells in contrast with the
absence of this monosialoganglioside in the plasma
membrane of F3II cells.
We then tested the effect of exogenous GM3 on the in
vitro growth of B16 and F3II cells. The addition of
GM3, at concentrations higher than 50 lg/ml, caused an
increase in the proliferation of GM3-positive B16 cells in
serum-free conditions. Conversely, exogenous GM3 in-
duced a significant growth inhibition of GM3-negative
F3II cells in the presence of 5% FBS (Fig. 2). Neither
cytotoxic nor proapoptotic effects were observed using
Fig. 1a,b Flow cytometry analysis of B16 melanoma and F3II
mammary carcinoma cells labeled with the anti-GM3 monoclonal
antibody M2590. a B16 cells were incubated with isotype control
antibodies (thick line), M2590 (thin line) or M2590 preincubated
with soluble GM3 (gray line); b F3II cells were incubated with
isotype control antibodies (thick line) or M2590 (thin line). Results
are representative of three independent experiments
Page 4
a non-radioactive cytotoxicity assay and a DNA frag-
mentation detection kit, respectively, suggesting a cyto-
static action of GM3 on F3II monolayers (data not
Serum antibody and lymphoproliferative response
of GM3/VSSP immunized mice
In order to assess the antigenicity of the vaccine, we
first studied the specificity of serum antibodies in im-
munized mice. The reactivity of sera was tested against
both the GM3/VSSP conjugate or unconjugated GM3
ganglioside. Mice of either C57BL/6 or BALB/c strains
were vaccinated with four doses of GM3/VSSP and
serum samples were obtained 7–21 days after comple-
tion of the immunization schedule. All mice immunized
with the GM3/VSSP vaccine developed IgG antibodies
against the GM3/VSSP conjugate. High titers were
obtained, reaching 32·106, and no significant differ-
ences were obtained between mouse strains. Interest-
ingly, animals immunized with the unconjugated carrier
VSSP plus Montanide developed similar titers against
the GM3/VSSP conjugate, suggesting that a strong
reactivity is directed against the carrier moiety of the
The binding of serum antibodies to ELISA plates
coated with purified GM3 was also assessed. Uncoated
plates were used as a control for non-specific binding.
Sera derived from all mice treated with the carrier pro-
tein plus Montanide or only with the immunological
adjuvant, as well as from untreated mice, contained
undetectable amounts of GM3-specific antibodies (IgM
and IgG titers <80). Analysis of serum samples derived
from C57BL/6 mice immunized with GM3/VSSP vac-
cine yielded the detection of low-titer GM3-specific IgM
antibodies in four of eight mice. Similarly, sera from
BALB/c mice showed detectable anti-GM3 IgM anti-
bodies in two of five mice. More interestingly, moderate-
titer anti-GM3 IgG antibodies were detected in both
strains, with titers reaching 1,280 (Table 1). In summary,
we found the presence of at least one class of antibody
against the purified GM3 ganglioside in five out of eight
C57BL/6 (62.5%) and in four of five (80%) BALB/c
immunized mice.
Next, we studied whether serum antibodies of im-
munized mice mediate direct recognition of tumor cells.
In a previous report, we showed that sera from C57BL/6
immunized mice have a strong reactivity against B16
melanoma using tumor sections from tissues embedded
either in Epon or paraffin (Alonso et al. 1999). Here, we
analyzed the reactivity of sera from GM3/VSSP immu-
nized mice against B16 and F3II tumor cells. Fig. 3
shows the serum reactivity of immunized mice against
these cells as analyzed by flow cytometry. Clearly, im-
mune sera recognized the surface of the GM3-positive
B16 cells while GM3-negative F3II cells were not
stained. The observed difference in serum antibody
binding was confirmed by immunoperoxidase treatment
of fixed cell monolayers. As shown in Fig. 4a, most
melanoma cells display a distinct positive staining using
sera from C57BL/6 immunized mice. Although the re-
activity was associated with the cell membrane, a diffuse
cytoplasmatic staining was also observed. In contrast,
there was no reactivity of immunized sera against the
surface of F3II mammary carcinoma cells. However, a
heterogeneous pattern of reactivity was found in endo-
membranes (mainly endoplasmic reticulum and Golgi
apparatus) in F3II cell cultures (Fig. 4b). In all cases,
there was no reactivity with sera obtained from un-
treated mice or mice receiving only the immunological
Preliminary results obtained in lymphoproliferative
assays showed an increase in the stimulation index of
Fig. 2 Effect of exogenous GM3 ganglioside on the in vitro
growth of F3II and B16 cells in the a presence or b absence of
serum. Data represent means±SEM (n=16). *P<0.001, ANOVA
Table 1 Specific anti-GM3 antibodies in sera of mice immunized
with GM3/VSSP. Mice were immunized with four doses of GM3/
VSSP vaccine and specific antibodies against purified GM3 gan-
glioside were detected by ELISA in 96-well microtiter plates coated
with GM3 as described in Materials and methods. All control mice
yielded undetectable levels of GM3-specific antibodies (IgM and
IgG titers <80)
Strainn Titer (n)
320 (1)
<80 (3); 80 (1);
640 (1)
<80 (4); 80 (1); 160 (2);
1280 (1)
<80 (2); 80 (2);
1280 (1)
Page 5
splenocytes derived from vaccinated mice exposed to
5 lg/ml of GM3/VSSP. Stimulation index in C57BL/6
immunized mice doubled the index of the control group
(9.7±1.7 versus 5.3±0.1). A similar response was ob-
served in BALB/c mice, in which case vaccination in-
creased the stimulation index by fourfold (20±1.0
versus 4.5±0.5).
Abolition of serum reactivity of GM3/VSSP vaccinated
mice against B16 melanoma cells by removal of cell
surface sialic acid
To establish whether sialic acid present in melanoma
plasma membrane plays a key antigenic role in serum
antibody response of mice immunized with GM3/VSSP,
we treated formalin-fixed B16 monolayers with neura-
minidase, a hydrolase that remove sialic acid residues
from the cell surface. Neuraminidase pretreatment
completely abolished the reactivity of serum from vac-
cinated mice against B16 melanoma cells (Fig. 4e).
Desialylation of paraffin-embedded tissue sections of
B16 subcutaneous tumors also inhibited humoral re-
sponse against melanoma cells (data not shown). The
possibility that protease traces contaminate neuramini-
dase preparations and may be partially responsible of
the described effect was ruled out. We tested the prote-
olytic activity of neuraminidase preparations using a
highly sensitive caseinolytic assay and no signs of
degradation were observed.
Relationship of in vivo antitumor activity induced
by GM3/VSSP vaccine with the presence of GM3
ganglioside on tumor cell surface
C57BL/6 or BALB/c inbred mice were vaccinated four
times with GM3/VSSP vaccine on days –63, –49, –35,
and –21, prior to B16 or F3II tumor cell challenge on
day 0, respectively. In both cases, syngeneic animals
received the minimal challenge to obtain a tumor inci-
dence of 100% (1·104B16 cells or 2·105F3II cells), as
indicated by preliminary experiments. As shown in Ta-
ble 2, preimmunization with GM3/VSSP vaccine sig-
inoculated with B16 melanoma cells. Vaccinated mice
survived longer than controls (Fig. 5a), and several an-
imals were sacrificed on day 90 after challenge with no
evidence of melanoma tumors. On the contrary, vacci-
nation failed to block tumor formation in BALB/c mice
challenged with GM3-negative F3II mammary carcino-
ma cells (see also Table 2 and Fig. 5b). Similar results
(not shown) were obtained inoculating smaller tumor
cell burdens, although tumor incidence was <50% in
control animals.
formation inC57BL/6
GM3 is a monosialoganglioside, being the most abun-
dant membrane glycolipid in melanoma cells (Hersey
et al. 1998; Nakakuma et al. 1992). Previously, we have
demonstrated that a GM3-based vaccine induced tumor
protection in mice challenged with melanoma cells (Carr
et al. 2001; Alonso et al. 1999). As with other vaccines,
the understanding of the underlying mechanisms in-
volved in immune responses is crucial to elaborate a
model of potential clinical application (Bitton et al.
In this work, we confirmed the expression of GM3
ganglioside in the surface of B16 melanoma cells using a
GM3-specific monoclonal antibody. Conversely, we ob-
GM3 on their surfaces, although reactive epitopes could
be detected in intracellular compartments. Other authors
and postulated that this microclusters take part in vital
cell functions, including adhesion, migration and, possi-
bly, cell signaling, by interaction of GM3 with c-Scr and
Rho transducer proteins (Iwabuchi et al. 1998b; Ya-
mamura et al. 1997). GM3 has been cited also as a regu-
latory molecule for epidermal growth factor (Meuillet et
al. 2000; Zhou et al. 1994), the basic fibroblast growth
etal. 1993). More recently,GM3 expressioninmelanoma
has been shown as a determinant of malignant phenotype
(Deng et al. 2002; Deng et al. 2000).
Exogenous GM3 stimulated the proliferation of B16
melanoma cells when cultured in serum-free conditions.
On the other hand, growth arrest induced by GM3
ganglioside in GM3-negative F3II mammary carcinoma
Fig. 3 Flow cytometry analysis of B16 melanoma and F3II
mammary carcinoma cells labeled with sera of GM3/VSSP
vaccinated mice. a B16 cells or b F3II cells were incubated with
control serum (thick line), or from vaccinated mice (thin line)


text:
A happy view of Daisy's House and Greet Daisy when it's not bathed in subjectivity
After you tour through Daisy's house, you'll come across her Film Silo in the rain. There you can observe Daisy finishing recording a scene from her celebrated film, "The Sorcerer's Dumpling. Be sure to pinch her fox!
You shouldn't try Princess Mode
© copyright 2011–2014
A Pants Aflame production


text:
PhotoTips.biz - (Free BW Photoshop Actions)
MyProFoto.com - (SmugMug Customization)
This week we head to the woods to focus on something that I like to call Landscape Detail Photography. I did not invent the idea, but that terminology may be my own creation. No matter, it is not exactly as easy as it sounds.
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Famous Idaho Potato Bowl comprehensive coverage: Aggies defeat Rockets, 41-15
Tom Smart, Deseret News
The Utah State Aggies have defeated the Toledo Rockets in the Famous Idaho Potato Bowl, the second win of USU's eight bowl appearances.
DeseretNews.com presents comprehensive coverage of this significant milestone for the Aggies.
Famous Idaho Potato Bowl: Williams carries Aggies to Potato Bowl victory
Brad Rock: Aggies take the next step in their journey to football prominence
Famous Idaho Potato Bowl: Young gun Keeton quarterbacks Aggies to victory
Notebook: USU assistant Mike Sanford accepts top job at Indiana State
Twitter recap: Utah State defeats Toledo 41-15 in the Famous Idaho Potato Bowl
Report Card: Williams, Aggies shine in Potato Bowl win
Postgame: Utah State defeats Toledo 41-15 in the Famous Idaho Potato Bowl
Halftime: Utah State leads Toledo 10-6 at halftime (+video)
Landon Hemsley is sports content manager for DeseretNews.com. Email: firstname.lastname@example.com Twitter: @EarlOfHemsley
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text:
Stock is up on higher than normal volume and no news as usual. QP buying ? shorts covering?
The volume was a little bit higher than usual, but the price hit close to it's highest closing price in a long time that had come with heavy volume. I hope that's a good sign.
Yes, its better than going up on light volume. Wind and wind related are all acting well though. Hard to differentiate its movement from wind(vestas ) in general but still good.
Sentiment: Buy


text:
A careful examination of frozen caribou poop has turned up two never-before-seen viruses.
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A study published today in the New England Journal of Medicine, to coincide with world Pneumonia Day, records the country's success in reducing pne... Read More
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The bed bug may be just as dangerous as its sinister cousin, the triatomine, or "kissing" bug. A new study from Penn Medicine researchers in the Center for Clinical Epidemiology and Biostatistics demonstrated that bed bugs, like the triatomines, can transmit Trypanosoma cruzi, the parasite that ... Read More
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Listed in: Food & Drink
5. Chemist Dad
A blog of father, chemist, husband that talks everything under the sun
Owner: asdomencil
Listed in: Personal
6. Proyection X Photography
las mejores pendejas encontralas en Proyection X : fotos, videos para descargar y mas...
Owner: nissneo
Listed in: Entertainment
7. shu84
Daily Inspirations of Photography, Fashion and more...
Owner: shu84
Listed in: Photo Blog
Tags: Fashion, Art, Inspirations, Design
8. Pal Raine
A personal blog talks about blog in general
Owner: Pal_Raine
Listed in: Personal
9. Starry Sky Ranch
I am a photographer and homeschooling mom to ten awesome kids. After years of ranching...
Owner: Kim_Fry
Listed in: Photo Blog
Tags: family, homeschool
10. XinYun @ SG
My thoughts and sharings, on my life, photography, events, tech gadgets and food.
Owner: xinyun
Listed in: Personal
11. Pans and Pots
Come and take a peek into my pans and pots. :) Let's cook together ;)
Owner: Zoka
Listed in: Food & Drink
12. Hobby Shobbys
Hobby Shobbys is all about hobbies of people around the world
Owner: lahorite
Listed in: Entertainment
Tags: Hobbies, cooking, collecting, outdoors
13. Views From An Urban Lake
A Wildlife and photography blog from Milton Keynes UK.
Owner: beolens
Listed in: Outdoors
Tags: Wildlife, Birds, Nature, Milton Keynes
14. The Taiwan Photographer
Photographs and information about Taiwan by Rich Matheson, a twenty-year resident of...
Owner: Matheson
Listed in: Photo Blog
Tags: Journalist, Professional, Taiwan, Travel
15. Kevin's Blog of Worthless Advice
Probably the only blog you'll ever want to read! Well, at least look at and enjoy...
Owner: Kevin_Hellriegel
Listed in: Humor
16. ApetitPois
ApetitPois is about exciting designs, beautiful spaces, gorgeous interiors, talented...
Owner: ApetitPois
Listed in: Lifestyle
Tags: inspiration, design, interiors, architecture
17. Huchchara Santhe
A travel blog, a photo blog, a nature blog, wildlife and birdwatching blog, trek and...
Owner: Santosh_Bengaluru
Listed in: Travel
18. Daily Poetry and Stories Portal
Daily Poetry Portal in English and Netherlands. Enjoy reading the Poems with amazing...
Owner: easybranches123
Listed in: Photo Blog
19. David Fabros Visual Arts
The official blog of photographer and writer David Fabros, his forays into light and...
Owner: Fabros
Listed in: Photo Blog
20. Wildlifist
Wildlife of California. Pictures of whales, eagles, bears, otters, foxes, and many...
Owner: Wildlifist
Listed in: Outdoors
21. Caramelos de Colores
A bit of my life, my hobbies and my art
Owner: Julieta_Andrade
Listed in: Personal
22. Timbobaggins Abroad
The travels pf Timbobaggins, wildlife enthousiast, photographer and guitarist. UK based.
Owner: Tim_Ward
Listed in: Outdoors
23. Elmer Loves Canon
A photo blog that features my pictures using my Canon EOS 1100D camera
Owner: elmerdomingo
Listed in: Photo Blog
Tags: Travel, Architecture, Foods, Buildings
24. Extra Mile Deals: Travel Promotions, Frequent Flyer & Hotel Rewards, and...
The Extra Mile Deals Blog informs consumers about the latest online deals, rewards...
Owner: xtramiledeals
Listed in: Travel
Tags: travel, hotels, airlines, destinations
25. Indonesia Dalam Kamera
Seeing the beauty of Indonesia through the camera and see the places of tourist...
Owner: dwindapratama
Listed in: Arts
27. Graphically Frank
Our creative realm.
Owner: JustCallMeFrank
Listed in: Arts
Tags: Paiting, Digital Arts, Mixed Media, mental health
28. Sugar Wanders
The Confessions of a Lonely Wanderer
Owner: jhbaloyo
Listed in: Travel
Tags: Travel, Leisure, Food, Entertainment
29. The Shadow Gamer
the Shadow Gamer providing the latest news and tips on video games, gadgets, internet...
Owner: Xingyi_Ho
Listed in: Games
Tags: video games, technology, gadgets, malaysia
30. Life with a Camera
Photographer working and living in West Yorkshire area. Weddings, Sports, Events, Travel
Owner: imaginepoland
Listed in: Photo Blog


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text:
Indian shares end up; autos gain, Reliance recovers
MUMBAI, Nov 1 (Reuters) - Indian shares provisionally ended higher for a second session on Thursday, led by gains in auto shares on higher-than-expected October vehicle sales while tech firm Wipro rose after it said it will separate its non-IT units to focus on outsourcing. Mahindra & Mahindra rose 2.18 percent, and Wipro shares gained 2.74 percent. Reliance Industries ended up 0.19 percent after falling 2.1 percent intraday. The BSE index provisionally rose 0.23 percent, while the 50-share NSE index added 0.45 percent. (Reporting by Manoj Dharra; Editing by Jijo Jacob)
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