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Victoria Street 03-34 Golden Landmark, 188061 How to get http://www.snemarketing.com Marketing & Sales Sne Capsule
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Historical Jesus 14: Last Supper
Why did Judas betray Jesus? What did Jesus teach his closest friends and followers at the last supper? Learn about the first "Lord's Supper" or communion as well as what was on Jesus' mind in his last hours before his arrest. He taught them how to carry on after he was physically gone and showed them how to serve one another by washing their feet. This tender dinner is the calm before the storm.
Course notes are available here.

subset: commoncrawl


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"I feel lit up by the idea. I'm very excited about becoming a dad and I'm preparing myself. I am just getting ready to be with a new little person and see what it is they want."
On what his life is like now:
"It is very calm. I look at the chickens, they hatch some eggs. Basically I'm like a village idiot — just looking at livestock. There is constant conflict between the primal drives: the drive to procreate, the drive to survive, and the drive to have status. But I am no longer deluded as to what may provide happiness. I'm so much happier over the course of the day to see one or two people and a few chickens, that's a good way of living."
On writing a book on addiction, due out next year:
"Addiction is about the way you relate to the outside world. If you're lucky, you'll get a substance misuse problem, which drills it right down. If you're not lucky, you'll have a food, sex or spending issue; those things are insidious because they're culturally endorsed, they're habitual."
On studying religion in global politics at the University of London:
"I am very interested in the role that religion and spirituality will play in the further formulation of world events and how secular societies are held together. The old idea [about politics] is dead and people don't know what's replacing it yet, so I'm spending some time discovering what it is."


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The two Dubai islands, The Palm Jumeirah and The Palm Jebel Ali, will be built in the shape of date palm trees and consist of a trunk; a crown with 17 fronds; and a surrounding crescent island – the back of which forms the breakwater. Collectively, the island will support more than 60 luxury hotels, 4,000 exclusive residential villas (sale), 1,000 unique water homes (sale), 5,000 shoreline apartments (sale), marinas, water theme parks, restaurants, shopping malls, sports facilities, health spas, cinemas and various dive sites.
Its Development
The project took four years of methodical planning and exhaustive feasibility studies in Dubai, ensuring that the islands would not disrupt the environment. The first stage of the development of The Palm Islands, includes the construction of the land foundations, which involves sand transfer and rock placement. Following this, the next stage will involve the building of infrastructure and services, as well as the 300 meter (990 feet) bridges linking the islands to the mainland. The final stage consists of the construction of homes, townhouses, and apartments.


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Planet Ballroom Dance Studio
Ponte Vedra
950 Marsh Landing Pkwy
Suite 330
Jacksonville Beach, FL 32250
Welcome to Planet Ballroom Dance Studio at Ponte Vedra and Jacksonville Beach; Jacksonville, Florida's premier Ballroom & Latin dance lesson experience! Come visit our beautiful, upscale and energetic ballroom dance studio located just off Highway 202 and A1A in Marsh Landing Plaza next to World Market!
Planet Ballroom Ponte Vedra offers the finest instruction in all Ballroom, Latin and Swing dances for couples, singles and teens! Learn to dance Tango, Waltz, Cha Cha, Salsa and Jive from our expert professional dancers! Beginners are welcome and partners are NOT required!
Come join the fun with us today! Call our award winning customer service team at (904) 595-5755 to schedule a complimentary orientation and dance studio tour! We'll see you soon on the dance floor!
Planet Ballroom Ponte Vedra serves the Jacksonville, Florida area including Jacksonville Beach, Ponte Vedra, Neptune Beach, St Augustine and south Jacksonville.
Franchisee: Tony Kantin
Tuesday thru Friday: 10am - 10pm
Saturday: 10am - 4pm


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Before Michael Ealy was famous…he waited tables at a high-end Caribbean restaraunt! What's moooore interesting is the variety of celebs that used to stop by and get their grub on. When Diddy ate there, his tab came out to 40 bucks. Watch this and find out how much he left for the tip:
Celeb Bad Boys Coalition [PHOTOS]
8 photos Launch gallery
Celeb Bad Boys Coalition [PHOTOS]
1. Chad Johnson1 of 8
2. DMX2 of 8
3. Suge Knight3 of 8
4. Chief Keef4 of 8
5. T.I.5 of 8
6. Katt Williams6 of 8
7. Lil Boosie7 of 8
8. Charlie Sheen8 of 8
comments – Add Yours


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Huh...I wonder! Rotovirus?
I took Rory and Alex to the pedi last Wednesday to get Rory's 4 month check-up and Alex's flu shot. All went swimmingly and I'm very proud of my boys.
One of Rory's vax was the rotovirus one by mouth. I know that you're supposed to wash your hands and all that stuff, blah blah blah (though I should point out that i was not reminded by the staff nor was I given the info sheet about it this time).
Anyway, flash forward to Saturday night. Alex is awake and throwing up. On into the next day. By Sunday evening his fever was low and it was broken by Monday. Vomiting stopped well before then.
Today Toby is down and out and I'm expecting the same thing to happen to him.
I was flipping through these cloth diapering FB groups and they're talking about using disposables after doing the rotovirus (you don't need to, hot washes and detergent kills it). And it hit me. THAT'S WHAT HAPPENED. I must have been lax in how I washed my hands or something. Not surprised about this really.
Oh well. Nothing to do about it. 'Cept give lots of water. Alex got 2 days of non-stop television, popsicles, and attention. Rory just got the vax for it so no worries there about him, lol.
So, ladies, as your babies get their rotovirus vax, remember to be THOROUGH in washing your hands and everything that your baby's poop may conceivably touch.
I could be wrong and this is an unrelated illness but it can't hurt to pass the warning on :)
Re: Huh...I wonder! Rotovirus?
I hope everyone is feeling better now!
Re: Huh...I wonder! Rotovirus?
Thankfully DH never developed a fever last night and he seemed to be feeling better in the wee hours of the morning when he got up to change out Rory's milk bag.
Alex is doing great but only peed like 2 times yesterday. He goes to daycare today but I'll be sending a large sippy cup with him that he's to drink out of constantly.
Re: Huh...I wonder! Rotovirus?
The ped did not give us any warning. Good to know.
Re: Huh...I wonder! Rotovirus?
They tell you to use sposies after the rotavirus just because the oral vaccine causes yucky diarrhea, not because of live virus contamination issues... I think the vaccine virus is too attenuated to spread like that... That's why they don't warn you. He could have picked up a random virus at the Dr office pretty easily too.
Re: Huh...I wonder! Rotovirus?
Yeah Tristan had nasty diarrhea for a day after he had the VAX
Re: Huh...I wonder! Rotovirus?
Well, Rory is sick now too. Ugh. Doc says it's a random virus. Good news is that Rory gained a pound in a week! He's 14lb and 8oz :-) 24.5cm in height :-) Haven't noticed any diarrhea.
Re: Huh...I wonder! Rotovirus?
I have first hand experience with the rotavirus. Hailey got it when she was 4 months old (two weeks after getting the vaccine). She woke up fine that morning and I nursed her. Then about 10 minutes later she threw up every drop that was in her belly. She did this twice. About 10-15 minutes later she started burning up bad. We waited it out a little bit to see if the motrin (yes I know I gave it to soon, I wasn't thinking) would kick in. During this time she had diarrhea and blood in her poop. Thats when I knew something was wrong. We took her to the ER and it was confirmed she had the rotavirus. She was given an IV and was in the hospital for 5 days. During that time she would not eat and so I had to pump to keep my supply up. I was at her side all but maybe 45 minutes the whole 5 days she was in the hospital. It was one of the worst times in my life :(
My Hailey bug during her hospital stay :(
Re: Huh...I wonder! Rotovirus?
:( poor baby!!!!! ^^ so scary. Rotavirus is not cool.
Re: Huh...I wonder! Rotovirus?
Poor thing! As awful and harrowing as the sickness sounds I'm looking at the pictures with her IV. When Rory got his IV placed when he got his tube they had to stick him multiple times in both arms and legs (and then 2 days later do it again). I can't see a baby with an IV and not get tears in my eyes, for both mom and baby. Thank God she is doing better!
I was relieved to find out that the live vaccine exposure and all our illness were just an unlucky coincidence. Don't kick yourself about giving motrin too soon, EVERY parent wants to ease their baby's discomfort ASAP.
Re: Huh...I wonder! Rotovirus?
She hated that IV board. It was so hard to watch her getting it. She was so blah that you could tell that she was not feeling well. Thank goodness she's been healthy ever since. I was so nervous about her getting the second dose and I'm nervous about Zack getting the vaccine all together. She's a thriving almost two year old now :)
Re: Huh...I wonder! Rotovirus?
Because your little girl obviously had bad time of it are they thinking about delaying the vaccine until 6 months just in case?
Re: Huh...I wonder! Rotovirus?
We haven't spoken about it yet. We see a new doctor tomorrow because their one we always saw in the office left to work closer to home. This doctor is still in the same office. I'll bring it up tomorrow when we go in.
When she was due for the second dose they did call the state agency who oversees the vaccine cases see if they should hold off on giving her the second dose because of her hospital stay but they said they felt safe giving it to her still.
Re: Huh...I wonder! Rotovirus?
Thanks for this info. Leif just got it Monday. How long do you have to be super careful?
Re: Huh...I wonder! Rotovirus?
I think up to 6 days but at least 3-4 :)
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Belonging Trailer (2004)
"The bittersweet bonds of love and betrayal" 12 September 2004 Drama, Romance, Foreign 96 mins
Jess (Brenda Blethyn) and town handyman Jacob (Kevin Whately) have been happily married 20 years, recently taking in a trio of Jacob's elderly relatives, including his mother (Rosemary Harris). Though the relations are demanding, kindhearted Jess -- who selflessly quit her job -- enjoys looking after them. But when Jacob disappears one day, Jess' life falls apart, and she must learn to cope with things on her own in this touching drama.
Watch the official Belonging 2004 trailer in HD below or find more Belonging videos on Vidimovie.


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decoracion, arte, bricolaje, casa de sueño, rincones especiales, muebles, interiores, espacios para niños, el hogar, casa confortables
For this weeks radio show, I was thrilled to take a break from home design- and human design, for that matter- to speak with the creator of my favorite line of dog designs: Lyndsey Drago of Billy Wolf. I never thought Id be a dog owner, but ever since adopting Hope, Ive been happier than I even knew I could be and Ive been buying little treats and accessories for her like crazy. One day at the vet, I asked Hopes doctor if a coat was necessary when the temperatures started to drop and she told me that some breeds (especially smaller dogs or dogs with short hair) need extra warmth if theyre outside walking in super-cold temperatures. I took that as a literal doctors permission slip to give into my crazy side and buy Hope a few coats for sub-30 degree days. The first and only brand of coats Ive gone back to over and over is Billy Wolf. Made from found, vintage and deadstock fabrics, Billy Wolf coats are chic enough to be worn by humans and functional enough to grow with your dog and keep them warm without getting in their way. I was thrilled when Billy Wolfs founder agreed to come into the city to talk with me on the show, so I hope youll enjoy her insight into designing for pets as much as I did. Thanks to Lyndsey (and her dog!) for making the trek from Montauk to Brooklyn for the show! [You can meet Lyndsey and check out her designs at the Brooklyn Night Bazaar and Renegade Craft Fair over the next few weeks!] xo, grace I think the best way to make eco-friendly products is to make quality products that will last. Its not something that youre going to throw away. [8:45] I want people to get something for their dog that they would want for themselves. [25:35] -Lyndsey Drago on After the Jump LISTEN: You can download the podcast on iTunes here, or stream it here on Heritage Radio


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The news that the footbridge from Marmion Terrace to Norham Road in Monkseaton is to be closed for six weeks for repairs really is almost unbelievable (News Guardian, July 10).
This footbridge is used daily by hundreds of people to connect Monkseaton to Whitley Bay and is an essential link.
They have already demolished the footbridge at Monkseaton stiation.
I am told a brand new footbridge could be made and fitted in a fortnight, is the Metro to dismantle the bridge then go on holiday?
Charles Hall


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Angostura Bridge
- Basic
- Data
- Media
- Publications
- Files
Angostura Bridge is a suspension bridge that spans the Orinoco River at Ciudad Bolívar, Venezuela. The Spanish word Angostura means "narrows". Built in 1967, the bridge has a main span of 712 metres. Until the opening of the Second Orinoco crossing 100 km downstream near Ciudad Guayana on 13 November 2006 it was the only bridge across the Orinoco.
Media2 photos
Building Activity
OpenBuildings added a digital referenceabout 6 years ago via OpenBuildings.com


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PHOENIX (AP) - Diana Taurasi married former Phoenix Mercury teammate Penny Taylor less than 24 hours before the team's season opener.
The 34-year-old Taurasi has played for the Mercury since 2004, helping the team win three titles. She also helped the U.S. win four consecutive Olympic gold medals.
The couple married Saturday.
Taurasi had a rough season opener on Sunday, shooting 1-for-11 in the 68-58 loss to the Dallas Wings.
Taylor, who will turn 36 next week, retired last summer after a 13-year career in the WNBA. She played her final 10 seasons in Phoenix and is now the team's director of player personnel.
She previously was married to a Brazilian beach volleyball player, but the couple divorced after three years. The two WNBA players had kept their relationship out of the public spotlight.


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Help with resolving a 1000 year old dilemma
I'm new to the forum and my query might seem very basic, but would appreciate an answer.
I came across the works of some medieval scholars who set to determine what is the minimum depth of water that extends across an area 10 cubits x 10 cubits that if you were to scoop water from it, the ground underneath it would NOT appear.
A cubit (elbow to fingertip) for the purposes of this calculation is taken to be 46.2cm. Hence, 10 cubits2 = (10 x 46.2cm)2 = a surface area of 213,444cm2.
I would appreciate if someone could answer what would be the minimum thickness for the film of water to hold across this entire surface?:confused:
|All times are GMT -4. The time now is 02:00.


text:
1. Urgent recall on Children's Panadol due to allergic reaction fears
Multiple batches of children's paracetamol products from Panadol have been recalled over contamination fears. [Yahoo News]
2. Global flooding 'will threaten billion'
A British aid charity is warning that by 2060 more than a billion people worldwide will live in cities at risk of catastrophic flooding as a result of climate change. [BBC]
3. For a Long Life, Don't Be Too Thin Or Too Heavy
'Today's overweight individuals have lower mortality than so-called normal-weight individuals'. [TIME]
4. High potato intake linked to high blood pressure
Researchers link increased potato intake to high blood pressure in adults. The study is only observational, but the findings are likely to spark debate. [Medical News Today]
5. Life expectancy worldwide has increased by 5 years, UN reports
Life expectancy worldwide has increased by about five years in the last 15 years according to a report issued by the World Health Organization in the fastest gain since the 1960s. [Fox News]
6. If You've Ever Had a C-Section, Probably Best to Avoid Home Birth, Experts Say
Pregnant women who've had a cesarean delivery in the past should not plan a home birth because they face a higher risk for complications, researchers warn. [Health.com]
7. FDA approves new drug to treat bladder cancer
A new drug to treat bladder cancer was approved by the U.S. Food and Drug Administration on Wednesday. [UPI]
8. Expecting Mothers Be Warned, Popular Pain Medication May Cause Birth Defects
A study, published in the journal Neurology, revealed that women who were exposed to pregabalin during the first trimester of their pregnancy were three times more likely to give birth to a child with birth defects. [Nature World News]
9. Can Most Cancer Really Be Prevented?
In a report published in JAMA Oncology, determined that lifestyle factors, like diet, smoking, and exercise, account for 20% to 40% of cancer risk. [TIME]
10. Bright light alters metabolism
Exposure to bright light could affect your metabolism, reports a new Northwestern Medicine study. [Science Daily]
Surprising discovery:
Your Brain Will Age Better If You Do This
One of the best ways to keep your brain sharp as you get older may be to stay busy, according to new research. [TIME]
#QuoteoftheWeek – "My own prescription for health is less paperwork and more running barefoot through the grass" ~Leslie Grimutter
Remember you can always reach us directly by emailing firstname.lastname@example.com or follow us on Twitter or on Facebook for more updates.


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WINDHAM, Maine (NEWS CENTER) –
A little over two weeks after fire tore through a popular Mexican restaurant in Raymond, families affected by the business' closing are getting some help from the community.
Friday night, a fundraising dinner will be held at the Windham Veterans Center with proceeds going to those families, waiting for the A La Mexicana restaurant to be rebuilt.
The dinner will feature a buffet, cash bar, raffle, live music and will double as a Cinco de Mayo party.
The event begins at 3PM and runs until 11PM with tickets sold at the door.
Copyright 2017 WCSH


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TCR hypervariable regions expressed by T cells that respond to effective tumor vaccines
- 770 Downloads
- Citations
A major goal of immunotherapy for cancer is the activation of T cell responses against tumor-associated antigens (TAAs). One important strategy for improving antitumor immunity is vaccination with peptide variants of TAAs. Understanding the mechanisms underlying the expansion of T cells that respond to the native tumor antigen is an important step in developing effective peptide-variant vaccines. Using an immunogenic mouse colon cancer model, we compare the binding properties and the TCR genes expressed by T cells elicited by peptide variants that elicit variable antitumor immunity directly ex vivo. The steady-state affinity of the natural tumor antigen for the T cells responding to effective peptide vaccines was higher relative to ineffective peptides, consistent with their improved function. Ex vivo analysis showed that T cells responding to the effective peptides expressed a CDR3β motif, which was also shared by T cells responding to the natural antigen and not those responding to the less effective peptide vaccines. Importantly, these data demonstrate that peptide vaccines can expand T cells that naturally respond to tumor antigens, resulting in more effective antitumor immunity. Future immunotherapies may require similar stringent analysis of the responding T cells to select optimal peptides as vaccine candidates.
KeywordsPeptide vaccines T cell repertoire Tumor antigens Peptide variants
Increased frequencies of cytotoxic T lymphocytes (CTLs), which recognize antigenic peptides from tumors presented by major histocompatibility (MHC) class I molecules, correlate with improved cancer patient survival [1]. CTLs and the effector molecules they produce are critical in the elimination of cancer cells [2, 3]; however, they are also subject to regulatory mechanisms. Tumor-specific CTLs often recognize peptides derived from tumor-associated antigens (TAAs). One important strategy for improving antitumor immunity and overcoming these self-tolerance mechanisms is vaccination with peptide variants of TAAs (also known as mimotopes, heteroclitic peptides, peptide analogues, or altered peptide ligands) that improve binding to either MHC molecules [4] or tumor-specific T cell receptor (TCR) molecules [5, 6]. These vaccines stimulate the expansion of the often low-affinity TAA-specific T cells that escape negative selection during development [7, 8] and are suboptimally activated by native tumor antigens [9, 10]. Clinical trials have shown increased frequencies of TAA-specific T cells following vaccination with peptide variants, demonstrating the potential of antigen-specific immunotherapy [11].
Peptide variants are most often selected as vaccine candidates because they bind with stronger affinity or stimulate TAA-specific T cells more effectively than the native tumor antigen in vitro. Paradoxically, vaccination with these variants often elicits T cells with diminished antitumor activity relative to T cells that naturally respond to native tumor antigens [12, 13, 14, 15]. Using the CT26 transplantable tumor model [7, 16], we previously showed that vaccination with variants of the immunodominant peptide, gp70423–431 (AH1), elicited variable antitumor immunity [17, 18]. These peptides robustly stimulated the tumor-specific T cell clone used to identify the variants and elicited more tumor-specific T cells from the endogenous repertoire of BALB/c mice than the native AH1 peptide [17, 18]. However, vaccination with these peptides elicited a range of antitumor immunity. The ineffective peptides elicited T cells that failed to kill AH1-loaded target cells or produce IFNγ after stimulation with the native tumor antigen [17]. Surprisingly, these functionally deficient T cells produced IFNγ after stimulation with the variant peptide used in the vaccine, suggesting that the ineffective peptide variants were not antagonists or partial agonists, as in other systems [19].
Understanding the mechanisms underlying the expansion of T cells that respond poorly to native tumor antigens is an important step in developing effective peptide-variant vaccines. The repertoire of T cells responding to peptide variants is often different than the repertoire of T cells responding to native tumor antigens [10, 12, 20, 21]. As addressed in this study, we hypothesized that the effective peptide variants likely elicited a different repertoire of tumor-specific T cells that expressed TCRs with higher affinity for the native tumor antigen. Since in vitro expansion of tumor antigen-specific T cells can dramatically skew T cell cultures in favor of higher-affinity T cell clones [22], we compared the binding properties and the TCR genes expressed by T cells elicited by peptide variants directly ex vivo. The T cells elicited by an effective peptide-variant bound fluorescent MHC-tetramers containing the native tumor antigen with a higher staining intensity. These T cells did not cross-react with an ineffective peptide variant, suggesting that the repertoires of T cells responding to these variant vaccines were different and did not overlap. In addition, the effective peptide variants elicited a repertoire of T cells that was closely related in sequence to the T cells elicited by the native TAA, expressing a common Jβ sequence and a common CDR3β motif, and the frequency of sequences containing this CDR3β motif correlated with tumor protection. Thus, the effective peptide-variant vaccines enhanced the expansion of T cells that respond to the native TAA, rather than activating a new subset of T cells. Importantly, these results demonstrate that T cells responding to native tumor antigens can be effectively stimulated to prevent tumor growth, provided they are activated through the TCR by the appropriate signal. The implication of these results is that the most effective peptide vaccines may be identified by measuring the responses of tumor antigen-specific T cells found in patients, rather than isolated T cell clones.
Materials and methods
Six- to eight-week-old female BALB/cAnNCr mice were purchased from the National Cancer Institute/Charles River Laboratories. All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee of National Jewish Health.
Sf9 and High Five insect cells ([23], Invitrogen) were cultured and infected as described [2, 17]. T cell clones were cultured as described [9]. The T cell lines were cloned by limiting dilution and expanded with 5 × 105/ml irradiated CT26 tumor cells expressing the costimulatory molecule B7 for 2 weeks prior to flow cytometry and TCR sequence analysis.
βgal (TPHGAGRIL [24]), AH1 (SPSYVYHQF [16]), A5 (SPSYAYHQF [9]), 39 (MNKYAYHML [18]), 15 (MPKYAYHML [18]), WMF (SPTYAYWMF [2]), and F1A5 (FPSYAYHQF [17]) peptides were identified as described. Soluble synthetic peptides were ≥95% pure (Chi Scientific).
Sf9 insect cells were infected with recombinant baculovirus (BV) encoding Ld and beta-2-microglobulin molecules covalently linked to peptides via a glycine-rich linker as described [2, 25].
Antibodies and staining reagents
Preparation of the fluorescent Ld tetramer linked to the AH1 peptide using a disulfide trap or loaded with exogenous AH1 peptide has been described [17, 18]. Soluble TCRs containing CDR3β motifs were prepared by inserting Vβ8.3 and Vα6 gene segments expressed by the A5-4E11 or 39-1D4 T cell clones into a BV expression vector [2]. Purified TCR molecules were conjugated to fluorescent streptavidin molecules as described and used to stain insect cells expressing membrane-bound peptide-MHC molecules [2]. The median fluorescent intensity (MFI) of Ld high cells was determined. Antibodies specific for the Ld molecule (28.14.8S), CD8 (53–6.7, Southern Biotech), IFNγ (XMG1.2, eBioscience), B220 (RA3-6B2, BD Pharmigen), CD4 (RM4-5, BD Pharmigen), and MHC-II (M5/114.15.2, BD Pharmigen) were used for flow cytometric analyses. The B220, CD4, and MHC-II antibodies were analyzed in the "dump gate" for the tetramer, IFNγ, and Vβ staining experiments. For costaining experiments, splenocytes from vaccinated mice were stained with Ld-tet linked to the AH1 peptide with a disulfide trap and conjugated to steptavidin-PE for 2 h at 37°C to facilitate internalization and limit competition between tetramers and Ld-tet linked to the F1A5 or WMF peptides conjugated to streptavidin-AF647 were added and stained for an additional 2 h at 4°C. All samples were run on a BD FACSCaliber flow cytometer.
Intracellular cytokine staining
Splenocytes were stimulated with the indicated peptide for 5 h in the presence of monensin, a protein transport inhibitor (GolgiStop, BD Cytofix/Cytoperm Plus Fixation/Permeabilization Kit, BD Pharmingen), fixed, permeabilized, and stained as described [17].
Tetramer decay and titration
Normalized fluorescence (N f) was calculated by dividing the T f at each time point by the T f at time 0. The natural log (ln) of the N f was plotted versus time and the data for each mouse were fit to a one-phase exponential decay curve [(y = y0*e(−kt) where y = ln N f, k = decay constant, and t = time)] using Prism version 4.0, Graphpad software. The half-lives were calculated for each mouse [(half-life = −ln(2)/k)] and compared using an unpaired two-tailed t test.
For the tetramer titration assay, splenocytes were stained as above for 2 h at 4°C. The MFI was calculated using the following formula [26]: MFI = (T f)/(% of AH1-tet+ of CD8+ cells). The MFI was plotted versus the concentration of tetramer and fit to a one-site binding curve [Y = (Bmax*X)/(K D + X)] using Prism version 4.0, Graphpad software. The K D values for each mouse were compared using an unpaired two-tailed t test.
TCR sequencing
Approximately 1 × 105 CD8+ AH1-tet+ splenocytes were separated with a MoFlo® High-Performance Cell Sorter. RNA was extracted using the RNeasy Mini Kit (Qiagen), and cDNA was synthesized using the QuantiTect Reverse Transcription Kit (Qiagen) according to the manufacturer's instructions. For the plasmid sequencing method, the cDNA was PCR-amplified using the primer sets described in Supplemental Table 1 [27, 28, 29]. Amplified DNA was cloned into the pCR2.1 vector using the TOPO TA Cloning Kit (Invitrogen), and sequences were determined using an internal Cβ primer (Supplemental Table 1) and capillary DNA sequencing instruments (ABI 3730s). For the high-throughput method, the cDNA was PCR-amplified for 15 cycles using a forward primer specific for all Vβ8 family members containing an adaptor sequence (in bold) and an internal multiplex identifier (MID) sequence (in italics, Supplemental Table 1) and a reverse primer specific for Cβ containing a different adaptor sequence (in bold, Supplemental Table 1). PCR fragments were separated by gel electrophoresis, purified using the Gel Extraction Kit (Qiagen), and further amplified 30 cycles using primers specific for the adaptor sequences. After gel extraction and quantification, the PCR products were combined and subjected to high-throughput sequencing as previously described [30]. Sequences from both methods were analyzed using a computer software program developed by our laboratory that uses a BLAST-type algorithm to identify Vβ and Jβ gene sequence information [28, 31]. This program translated and aligned the sequences, identified the MID sequences and sorted the results according to each vaccine, distinguished the germline-encoded and randomized CDR3β region of each sequence, calculated the length of each CDR3β region, and determined the number of sequences containing the shared CDR3β motif. The sequences of the TCRs expressed by the T cell clones (Fig. 6a) were determined by directly sequencing PCR products amplified from their cDNA using the Vβ8.3 and Cβ primers or Vα6 primers.
Vaccination with the F1A5 peptide elicits T cells with higher affinity for the AH1 peptide
Previously identified peptide variants effectively stimulated a tumor antigen-specific T cell clone both in vitro and in vivo, but elicited variable antitumor responses from the endogenous T cell repertoire [17]. Vaccination with the F1A5 peptide variant protected 90% of mice from tumor growth, while tumors grew in all of the mice vaccinated with the WMF peptide [2, 17]. The increased tumor protection afforded by the F1A5 peptide was attributed to the expansion of more tumor-specific T cells that exhibited effector function after stimulation with the AH1 peptide, although the mechanism for this differential expansion of functional T cells was not determined [17].
Since tetramer-dissociation rates correlate with peptide-MHC/TCR affinities in some systems [26], we measured the half-life (t 1/2) of tetramer binding to the responding T cell repertoire (Fig. 1b). The average t 1/2 of T cells from F1A5- and WMF-vaccinated mice was not statistically different (Fig. 1biii). Although many factors contribute to K d values calculated using multivalent interactions [35], these observed avidity differences may be due to faster on-rates rather than longer half-lives [36]. In summary, the F1A5-elicited T cells bound to the AH1 antigen with increased affinity, which may explain the previously described improved effector function exhibited by T cells responding to this peptide vaccine [17].
Few tumor-specific T cells cross-react with both the F1A5 and WMF peptides
In some systems, functional measurements more accurately assess the number of antigen-specific T cells [37]. To ensure the tetramer-binding experiments did not underestimate the number of T cells that cross-react with both peptide variants, we analyzed intracellular IFNγ production by splenocytes from vaccinated mice stimulated with either the F1A5 or the WMF peptides (Fig. 2c, d). Similar to the tetramer staining results, few T cells cross-reacted with both variant peptides. In addition, as previously reported [17], few WMF-elicited T cells produced IFNγ after stimulation with the AH1 peptide (Fig. 2d). Together, these data suggest that vaccination with the F1A5 and WMF peptides elicited different portions of the endogenous AH1-specific T cell repertoire.
F1A5 and WMF peptides elicit different repertoires of T cells
To directly characterize the TCR gene usage of the AH1-specific T cells elicited by the peptide variants, we co-stained splenocytes from vaccinated mice with the AH1-tetramer and a panel of antibodies to the mouse TCR Vβ genes (Supplementary Fig. 2a). In addition to the F1A5 and WMF peptides, we also analyzed the response to the native tumor antigen and to three other previously characterized peptide variants, 15 (ineffective), and A5 and 39 (effective) [9, 17, 18]. All of these peptide variants were also selected using the Vβ8.3+ CT T cell clone and, not surprisingly, they elicited AH1-specific T cells that predominantly expressed the Vβ8 family of TCRs (Supplementary Fig. 2b).
Effective and ineffective peptides elicit different repertoires of AH1-specific T cells
Surprisingly, although these junctional sequences were similar to that expressed by the CT T cell clone (Fig. 6a), we did not find the exact CT-TCR sequence in these mice or in the mice vaccinated with the peptide variants. The absence of the CT T cell clone in this study suggests that this clone is not representative of the endogenous repertoire of T cells responding to these vaccines and explains the lack of cross-reactivity between the F1A5- and WMF-elicited repertoires (Fig. 2). Perhaps the different mode of immunization (whole cell irradiated CT26-GM [9]) and the in vitro expansion used to generate the CT clone accounts for its dominance in those studies, and absence here.
TCRs containing the CDR3β motif bind to the WMF peptide with lower affinity
To study the binding properties of the motif-containing T cells described above, we required both the β- and α-chain sequences of the TCRs identified in Figs. 3 and 4. We expanded and cloned AH1-specific T cells from vaccinated mice as described [9]. The TCR genes expressed by these T cell clones did not reflect the sequencing results we obtained from fresh uncultured splenocytes, particularly for the T cell clones from mice vaccinated with the ineffective peptide variants (Fig. 6a). However, we obtained several antigen-specific T cell clones that expressed TCRs containing the CDR3β motif and that had identical Vβ chains as some of those identified ex vivo. These T cell clones expressed a Vα6 TCR, distinct from the Vα4.11 TCR expressed by the CT T cell clone (Fig. 6a).
To determine the binding properties of TCRs containing the CDR3β motif, we subcloned the TCR molecules expressed by the A5-4E11 and the 39-1D4 T cell clones (Fig. 6a) into a BV expression vector. The sequences of these TCRs were highly selected in mice vaccinated with AH1, A5, and F1A5 peptides and not identified in mice vaccinated with the ineffective 15 and WMF peptides. We previously showed that the relative binding of multimeric-soluble TCRs to peptide–MHC complex expressed on BV-infected insect cells correlates with the affinity of monomeric TCR for the peptide–MHC complex determined by surface plasmon resonance [17, 18, 33, 39, 40]. Therefore, we stained insect cells expressing each of the peptide variant–Ld complexes with fluorescent multimeric TCRs derived from the A5-4E11 and 39-1D4 clones. The soluble TCRs bound to insect cells expressing the native tumor antigen, but not the irrelevant βgal peptide, demonstrating that these TCRs are specific for the AH1 peptide (Fig. 6b, c). The staining intensity of insect cells expressing the WMF peptide was significantly lower, indicating that this CDR3β motif-containing TCR does not bind efficiently to the WMF peptide (Fig. 6b). The binding of the 39-1D4 TCR more closely correlated with its representation in the ex vivo repertoire analysis, binding to the A5 and F1A5 peptides and not to the WMF peptide (Fig. 6c). The low-affinity interaction between the CDR3β motif-containing TCRs and the WMF peptide provides a simple mechanistic explanation for the absence of these cells following vaccination with the WMF peptide.
Although the peptide variants used in our study effectively stimulated a tumor-specific T cell clone in vitro, they elicited variable responses from the endogenous T cell population. In this study, we found that effective cancer peptide vaccines stimulate T cells with hypervariable regions similar to those that respond to the natural tumor antigen, not a de novo T cell repertoire. These results suggest that the heterogeneity in the repertoire of AH1-specific T cells contributes to the ineffectiveness of some of the peptide-variant vaccines.
Similar to our results, a study of the T cell response to the Melan-A/MART-126–35 TAA showed that the TCRβ gene usage of the T cells responding to the native peptide was broader than that responding to the peptide variant [21]. They also identified a CDR3β motif that was shared among several patients, a so-called public TCR, although these motif-containing T cells were not among the dominant clones identified in each patient and did not have a higher functional avidity [21]. They concluded that the repertoires responding to the peptide variants and the native tumor antigen were subtly different but over-lapping, and that the observed functional differences were due to subtle structural changes in the TCR. In contrast, the T cells responding to the ineffective peptide variants in our study express a unique repertoire of TCRs that do not overlap with the repertoire responding to the native tumor antigen and that do not contain a significant fraction of the shared CDR3β motif. Therefore, in our study, only the effective peptide variants elicited a repertoire of T cells that over-lapped with those responding to the native tumor antigen.
The most effective peptide variants (A5 and F1A5) have minimal amino acid sequence changes relative to the AH1 peptide. It is, therefore, not surprising that these vaccines raise similar T cell repertoires. These data suggest that peptides with conservative amino acid changes may be more effective in stimulating antitumor immunity by eliciting a repertoire of T cells that mimics the response to the native tumor antigen. In agreement with this premise, another study concluded that substitutions at MHC-binding positions create immunogenic peptide variants with an overall similar structure to the native peptide [4]. However, the opposite has also been shown in these studies. Peptides specifically designed to encode conservative amino acid changes were ineffective vaccines and raised a repertoire of T cells different from that of the native tumor antigen [10, 20, 41]. These discrepancies suggest that even subtle amino acid substitutions may cause unpredictable changes in the repertoire and binding kinetics of the responding endogenous T cells.
The T cells responding to the F1A5 peptide express higher-affinity T cell receptors that functionally recognize the AH1 TAA better than the T cells responding to the ineffective peptide, WMF. Furthermore, soluble TCRs encoding the common CDR3β motif do not bind efficiently to the WMF peptide, providing a mechanistic explanation for the lack of these TCRs in the responding repertoire. We predict from these data that the structure of the WMF peptide prevents productive interactions with TCRs containing the common CDR3β motif. Specifically, the tryptophan residue at position 7 of the WMF peptide may affect the position of the adjacent tyrosine residue at position 6, precluding its predicted interaction with the tyrosine residue encoded by the Jβ2.6 gene segment of the motif-containing TCRs. Important for peptide vaccine development, these results imply that T cells responding to the ineffective variants (like WMF) cannot be "rescued" or forced to respond to the tumor antigen with better adjuvants; they are not anergic or improperly activated, they simply express T cell receptors that interact poorly with the native tumor antigen.
A clinically relevant strategy in the design of effective peptide vaccines is to optimize the T cells used in the identification of peptide variants. We selected our peptide variants based on improved binding to and activation of the CT clone, an AH1-specific T cell clone that was in vitro expanded and is not similar to those analyzed ex vivo (Fig. 6). The T cell clones that were found in the endogenous repertoire bound to the peptides that elicit better tumor-specific responses (Fig. 6). Therefore, we predict that peptide variants selected with an ex vivo tumor-specific repertoire, rather than an in vitro selected T cell clone, may be the most effective peptides for cancer vaccines.
Due to the unpredictable nature of the responding T cell repertoire, even if optimal T cells are used for peptide selection, ineffective peptides may be unavoidable in peptide-variant vaccine development. Therefore, a general strategy to improve the response to peptide variants such as the WMF peptide should be considered. In this particular case, rather than providing additional adjuvants or cytokines during T cell activation, the repertoire of the T cell response needs to be re-focused to those with high affinity for the native tumor antigen. Although strategies to engineer T cells with receptors that have high affinity for tumor antigens have been developed and employed in the clinical treatment for cancer [42, 43], effective antitumor immunity may be more easily achieved by "boosting" the peptide-variant responses with native tumor antigens [14, 44]. This strategy may increase the number of high-affinity T cells that cross-react with native tumor antigens and reduce competition with T cells that only bind to peptide variants.
In summary, peptide-variant vaccines improve the proliferation and activation of T cells that respond to native tumor antigens. Because the T cell response is unpredictable and different for every antigen, optimal peptide variants may be most successfully identified using a representative TAA-specific T cell population to select preferred residue changes from a large and random set of peptides.
This work was supported by NCI CA109560 and ACS RSG-08-184-01-LIB. K.R.J. and J.D.B. were supported in part by the Cancer Research Institute Predoctoral Emphasis Pathway in Tumor Immunology Fellowship.
Conflict of interest
The authors have no conflicting financial interests.
Open Access
This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
Supplementary material
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While Viognier is one of the classic grape varieties in the Rhone valley of France, its introduction to Washington State has been relatively recent. Ciel du Cheval decided to plant three acres of Viognier in 1994. The vineyard was looking for another white varietal that would thrive and produce a wine of distinct character. It was an immediate success. Starting with the first harvest in 1997, Viognier from the vineyard proved to be outstanding. In 2000, Ciel du Cheval expanded its Viognier planting.
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A pure and fesh experession of Viognier. A lively wine, it has wonderful floral and spicy aromas. The palate is fullbodied and it has a long finish.It was fermented in stainless steel, so there is no oak to cover fruit. Learn more ...
2012 Production Team
A grower owned winery, Côtes de Ciel was founded in 2012 by the Holmes family of Ciel du Cheval Vineyards. At Côtes de Ciel, we believe that all wine starts its journey in the vineyard and moves through the winery. Throughout this process, many people make a significant contribution and impact on the wine. Below are a few of the key people that impacted the 2012 vintage.


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About Us
Klean Carbon is part of the fast growing Klean Group of companies that originated in Tokyo, Japan, and is now headquartered in beautiful Vancouver, Canada. Initially focused on renewable energy, resource recovery and recycling sectors, the group has since diversified its portfolio and is now the involved integrated energy solutions, clean technologies, commodity production and distribution, environmental engineering, construction, finance, management services and consulting to businesses and governments around the world.
Klean Carbon® is a venture of Klean Industries Inc, into manufacturing of high-quality and environmentally superior recovered carbon black (rCB) know as CBk. Klean Carbon® CBk-EX®, CBk-RX® and CBk-SX® are new grades of carbon black reinforcing fillers that can fully or partially replace N300, N600 and N700 series of traditional Conventional Carbon Blacks "CCB".
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CBk is a green alternative to virgin carbon black and was established in response to the growing problem of carbon waste and carbon emissions. Understanding the global impact of transforming carbon waste (emitted from point sources such as refineries, chemical plants and electric utility plants) into usable energy, the Klean Team has aggressively developed new technologically advanced solutions to curb the carbon pollution problem and has quickly established a strong presence in major metropolitan markets in North America and Europe
By delivering a new form of carbon transformation, Klean has produced varying grades of recovered carbon blacks and activated carbon by developing technological services to help customers and society meet health, safety and environmental challenges related to product development and recycling.
Carbon Transformation: The Klean Advantage
Klean transformation solutions offer superior cost-to-transformation ratios and a viable, profitable and sustainable manner in which to solve the global problem of carbon pollution.
Feedstock Waste Carbon: A Global Advantage
Klean addresses the concerns of carbon pollution and creates new sources of carbon-based commodities by securing and transforming carbon waste feedstock.
Klean's co-combines the production of new raw materials and renewable energy from carbon waste feedstocks:
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Kason Fined For Customer Allocation Plot In Canada
The Ottawa court issued the CA$250,000 ($260,392) criminal penalty against Kason on March 8 for its role in the customer allocation conspiracy with Component Hardware Group Inc., which has already forced the two hardware makers to pay out millions to settle with U.S. prosecutors and private litigants alike.
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The BOA™ Band provides circumferential pressure on the upper extremities for the purpose of obtaining peripheral venous access. Once the device is applied, it is rolled downward forcing blood into the veins making them more visible and palpable.
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Proven Safety Tips For Backpacking In Europe
Personal safety is often a major concern for people backpacking across Europe, more so for first time backpackers. Europe as a continent offers breathtaking experiences, ranging from the majestic cathedrals of Paris to the plummeting coastline of the Amalfi. So, all one needs besides a few essentials is an adventurous heart. For instance, you need to know what to pack, how big your backpack ought to be, as well as how to find your way around. Whilst some alarming and unusual crimes have been reported by backpackers in Europe, you need to first and foremost protect yourself from simple thefts.
To begin with, you need to follow a raft of simple strategies to protect yourself, secure your belongings, and make backpacking through Europe an unforgettable experience. The most basic tip, of course, is that you should never walk around any European destination at night minus a friend or close acquaintance. You may as well have to carry some kind of personal safety device, such as a Coleman brass whistle or pepper spray can, for use in unpleasant and emergency situations.
Backpacking Essentials – You need to have a valid passport though most of the European countries never ask for visas if one has a valid U.S one. It's also advisable that you get yourself a Eurail Pass before you venture out as it would afford you unlimited travel through a total of 17 European countries. Buying passes on the go is very much expensive. Additionally, don't forget to carry at least one ATM, credit card, plus your service numbers just in case of loss.
The other most vital safety and security gears that you can bring along are an assortment of small combination locks plus keys. Remember that it's much easier for a thief to access your backpack and steal just a few things while you're making a purchase or talking on the phone than for him/her to dash off with the whole pack. So, you should see to it that you lock all the backpack's zippers, especially those containing valuables, to avert this.
Although napping on a train is a good way to get some much needed rest, it presents such an opportune moment for thieves to steal your backpack without your knowledge. It's therefore much more practical to secure your backpack to a train's luggage rack using a larger lock. You may as well have to opt for those hostels that usually offer rental lockers to patrons to protect their valuables at night or while they are out.
Items like cameras or mobile phones are probable targets for thieves, credit cards and hard cash are normally what crooks want. You should thus keep all your vital documents, such as airline tickets, rail passes, and passports in an elegant money belt or waist pack. Money pouches or money belts are far much safer than old-style fanny packs as they are worn closer to the body, making it harder for somebody to steal them.
Be very careful about the amount of hard cash you carry around with you. Major stores and restaurants in European cities accept debit or credit cards, plus you can even make arrangements to book your hotel through reliable sites. Nonetheless, be sure to check with your local bank about international ATM usage fees.
All in all, going backpacking in Europe will definitely offer you lots of tourist spots spread all over the continent. You will get to visit awesome cities, like Rome, London, and Paris, plus a host of other sites like Trevi Fountain, Louvre Museum, Eiffel Tower and Buckingham Palace.


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- Company:
- Elan Emerging Technologies Pvt Ltd
- iBzilla – Bugzilla on iPhone
- Version:
- 3.11
- Post Date:
- 2018-01-16 15:16:48
- Price:
- Free
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For the past few months I have been lucky enough to work with the team that was integrating LotusLive and SugarCRM. We are officially announcing availability of this integration at Lotusphere 2011 here in Orlando, FL. As part of the hard work that everyone has put into this effort I wanted to ensure that at least one SugarCRM engineer who worked on this integration was present for the "festivities." Lucky for me I was able to attend.
The theme of Lotusphere 2011 is "Get Social. Do Business." and as part of the kickoff Kevin Spacey, yes Kevin Spacey the actor, was the guest speaker. His production studio, TriggerStreet, was behind the movie "The Social Network." And as part of that effort and his quest to continue to find more screenplays, he has used social networking to try to reach out to more people. I have to say sitting in a crowd 10,000+ and watching Kevin Spacey was definitely quite a gift. But we're at Disney World so this should be the norm right?
I have never been to a tradeshow where I was on the other side of the booth and so this was quite a treat. Even more so because I got to see the types of questions people ask from: "What is CRM?" to "What makes SugarCRM stand apart?" I think it was sometimes refreshing to people when I said "I am an Engineering Manager so you will be hearing things from that perspective from me." The two most common themes amongst the questions would probably be
- I want my CRM to be simple and intuitive
- How can I integrate with my existing systems
This has been such a useful from an engineering perspective to see what people are concerned about. At the end of the day they care about what everyone cares about. I want to do things quickly and easily and it should be simple to understand.


text: 0 0 Stucco dogs playing this Friday June 17th.Join Us at spice bistro.Call to make reservation 801-930-9855 and get free appetizer. Chefs trying new recipes for summers at Spice Bistro.Join us for Patio Dining or takeout 801-930-9855. 0 Comments Leave a reply Click here to cancel the reply Your email address will not be published. Required fields are marked *CommentName * Email * Website Notify me of follow-up comments by email. Notify me of new posts by email.


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Download EzyDVD apk 1.0.6 free for Android smartphone
Posted by Élan Media Partners in Shopping | July 26, 2015 | 11 Comments
Apk file size: 1.2 MB
EzyDVD is Australia's largest online DVD and Blu-ray store, and the official EzyDVD app for Android brings you the online shopping experience directly to your mobile device.
The EzyDVD app for Android allows you to browse through our extensive range of Blu-rays, DVDs and Games, keep track of coming soon and new release titles, use the barcode scanner to look up pricing and availability for thousands of titles, and make purchases safely and easily.
Creating an EzyDVD account allows you to build your personal wishlist and share with friends, stay informed of our amazing offers, and receive updates for your online orders.
Too Ezy!
Whats new
Security updates
Minor improvements
Élan Media Partners part of our Shopping and have average installs from 1000 to 5000. Last Update July 26, 2015. Google play rating is 75.2727. Current verison is 1.0.6. Actual size 1.2 MB.
Similar applications
CatchOfTheDay: Online Shopping2899 Comments
11 Comments
Robert Byrne
Search function poor Can't find Heroes series DVD with app. Can from web site.
Very little content Doesn't seem to have a lot within the app why b does it not have everything that ezydvd has on their web site?
Andrew Janson
Unable to scroll Unable to scroll on Samsung Galaxy S2 - please fix.
Anthony Stratton
Finally Great app, really helpful.
Julia Salter
Easy to use Great range of product easy to find what I'm looking for- love the barcode scanner tool!
Ngoc Nguyen
Very Nice apps Cool, simple, and functional. Really love the blue. Exactly what I want. Truthfully I have been waiting for this for years. :-) <3 love it!!!
Frank An
Very handy app. Nice work
Todd Armstrong
Great app, runs smooth. I love getting my games from EzyDVD and it just got Ezyier
Lima Lima
Won't install waste of time
Tony Chirinian
Error message Unable to retrieve cart??
Kylie Jackson
I lovec this application


text:
Why Am I Afraid To Tell You Who I Am
first edition
by POWELL, John
IL: Argus Communications, 1969. Very Good. First edition. Very good in wrappers. Paperback is lightly rubbed at spine ends and corners, lightly soiled. Please Note: This book has been transferred to Between the Covers from another database and might not be described to our usual standards. Please inquire for more detailed condition information. (Inventory #: 221246)


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Impeached South Korean leader pulls out of questioning over corruption scandal
South Korea's impeached president has cancelled plans to undergo questioning in protest at leaks to the media about the investigation, according to p rosecutors.
Park Geun-hye had agreed to be questioned by prosecutors on Thursday over her alleged involvement in the corruption scandal that led to her impeachment.
Prosecutors said both sides had agreed not to disclose the time and place of the questioning until it was over.
Prosecution spokesman Lee Kyu-chul said Ms Park's lawyers told his team they were pulling out of the questioning after accusing prosecutors of leaking information about it.
The accusation came after SBS TV reported on Tuesday that Ms Park would be questioned on Thursday at the presidential Blue House.
Mr Lee said prosecutors did not leak the information.
The sudden rejection of what would have been the first prosecutorial questioning of a sitting president is the latest in a series of stand-offs between Ms Park and prosecutors digging into her alleged involvement in the biggest political scandal in South Korea in decades.
The scandal saw millions take to the streets in protest and led to Ms Park's impeachment in December.
The Constitutional Court is holding a trial to determine whether to formally end her rule or reinstate her presidential powers.
Mr Lee said prosecutors still want to question Ms Park but nothing has been determined about when it might happen.
Ms Park faces allegations that she let a long-time confidante, Choi Soon-sil, pull government stings from the shadows and extort an enormous amount money from companies in collaboration with presidential officials.
Choi and several of Ms Park's former presidential aides have already been investigated over the scandal.
By South Korean law, a president has immunity from prosecution while in office, except in cases of grave crimes such as treason.
Ms Park's officials have also blocked prosecutors from searching the Blue House.
Press Association


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what do you guys think about a guy wearing panties? part of it is for erotic but other part well I don't know it feels good. I'm debating whether or not to go buy a pair and I don't know what style to get or where to buy them.
Most Helpful Guy
np with me.. it's yer life bro 8)10|10|0


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2016 Toyota 4Runner in Birmingham, AL
Drivers shopping for a new SUV are looking for a vehicle that is ready to handle whatever situation and terrain it may encounter. These drivers are often adventurous and need a vehicle that will be able to keep up with their lifestyle. SUVs are also perfect for drivers needing more space for their family. These vehicles are spacious and versatile so the whole family can fit comfortably without having to result to a minivan. One of the most popular choices is the 2016 Toyota 4Runner. This SUV is tough and has impressive capabilities.
The 2016 Toyota 4Runner starts out at $33,510 with an estimated 17 mpg in the city and 22 mpg on the highway. Depending on the model, the 2016 4Runner can fit 5 or 7 passengers. Drivers needing more passenger space can choose a model with an optional third row. If there is not a third row, the second row is equipped with a 40/20/40 split folding feature. Every 2016 Toyota 4Runner comes with the Star Safety System and ToyotaCare, which features a no cost maintenance plan and roadside assistance.
|4.0-liter V-6 engine producing 270 horsepower
|Heated exterior mirrors with available turn signals
|Available TRD Pro trim with unique features
|Integrated fog lights
|In-key remote keyless entry
|17 or 20-inch alloy wheels
|5,000 lb. towing capacity
|Back up camera with projected path
|Available third row seating
|Daytime running lights
|6.1-inch high-resolution touch screen display
|Power rear glass window
Test drive the 2016 Toyota 4Runner in Birmingham, AL
All trims come set with Entune Audio featuring MP3, USB ports, Bluetooth and XM Radio capabilities. There are available upgrades for the Entune Audio system as well. Each model features a multi-information display which shows the outside temperature, a clock, the average fuel economy and distance until empty. The 2016 Toyota 4Runner allows drivers to stay in tune with their vehicle with standard features like a tire pressure monitoring system. Drivers looking to upgrade the 2016 Toyota 4Runner can choose one of the higher trims which feature an optional sliding rear cargo deck with under-floor storage, a power moonroof, available push button start and so much more!
Those interested in learning more about the 2016 Toyota 4Runner are encouraged to contact our team at Serra Toyota for more information. We're sure drivers will love this new SUV because it's versatile, full of technology and ready for all terrains. Schedule a test drive and be one of the first drivers behind the wheel of the new 2016 Toyota 4Runner.


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Pa Knowledgebase Plugin Wordpress Plugin is used by 1 websites. Among 1, 0 site is within top 10,000 and 0 site is within top 100,000 websites.
Wordpress Plugin Information of
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Top 1 Websites built with Wordpress plugin
Social Figures – Social Media Monitoring


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Wedding Minister in and around Magdalena, Nm.
Life's Moments are those times in life that we decide to make a special recognition of life altering occasions. Weddings are celebrations, weddings are ceremonies, but most importantly weddings are a new beginning, a moment in time to be treasured. Having the best Minister is important.
For these minutes, we provide thorough Wedding Minister solutions, Wedding officiating, and also wedding celebration services. From the very conventional to modern services, our wedding events and also officiating solutions are designed to reflect your wants, desires and also dreams. We will assist you to craft an occasion, spiritual or nonreligious that will certainly be a cherished memory for many years to come.
Pastoral solutions and also wedding celebration officiating in the Magdalena, Nm location is our specialty, but we could travel if the requirement arises. We provide a full array of Wedding packages; from elopement on the courthouse steps to the most elaborate formal wedding you might want.
We also provide the finest in Justice of the Peace or nonreligious weddings. These civil ceremonies provide their own kind of celebration, whether it be with family members or just a couple and also a couple of witnesses, your wedding celebration is an unique minute to be valued.


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Having completed our Umrah-e-Tamattu, the next few days in Makkah were for us to make the most of and schedule as we wish. We were only required to be at our group lectures that would prepare us for the upcoming Hajj, and so our routine, the one Asif and I had kind of unintentionally accustomed to, was going to be quite the same until Friday.
I woke up around 3:30am as we had planned to meet for 4am in the lobby and head to the Haram prior to Fajr prayers. Today, we decided to go all the way to the rooftop. The crowds were still not as much as people were still arriving in Makkah, and so we had a good chance of watching the Tawaf from above. We sat a while, prayed Fajr, and then made our way to the railings to see how beautiful the view of the Holy Ka'ba was from the top.
As more people arrived, we made our way out of the Haram and went back to the hotel. In the afternoon, we didn't go to the Haram for Dhuhrain prayers as both of us were feeling under the weather, with the intense heat! There were many who didn't dare to venture out in the scorching heat in the afternoons and prayed in the common lounge. In order to keep well and not overdo it, we decided we would do that too.
The afternoon lecture today was by Abbas Mama on the importance of Makkah and the benefits of being in the city. Habib uncle then ran through logistical details, followed by lunch. After lunch, we took time to rest.
At around 5pm, I met Asif again in the lobby and we went back to the rooftop of the Haram. We were fortunate to have the opportunity to recite Du'a-e-Mujeer whilst looking at the Holy Ka'ba. I can't really describe the feeling but it was so peaceful. No worries of the world or life back home would come into your mind, and you were solely focused on where you were at that moment. We prayed Maghribain prayers and made our way out of the Haram. Each time we had to leave the Haram, we felt like we wanted to stay a little longer. We were never really ready to leave, and after that first day we didn't really ask each other 'are you ready?' Asif would always say 'you can't really be ready to leave here, can you?' and so reluctantly one of us would say 'we should head back now…'
We stopped by a mini market place on the way back just to see what was situated around the Haram. It looked like a nice place to maybe get a few souvenirs for family back home. We didn't want to spend too much time shopping, so we decided to allocate two hours maximum in the remainder of our trip to get a few gifts. Our timing was good, as the adhan for Eisha began, all shops began to close and we headed back to the hotel. It was strange to us to see that people had 'Wi-fi' connection in their prayer and were praying Jama'at with those in the Haram. In the middle of the streets, you'd see clusters of people who were following along with the Imam.
We got back to the hotel in time to hear a little of the lecture that was being given by Shaykh Haider Shirazi, and then had dinner. I decided on an early night, and by 11pm, I was deep in sleep! Tomorrow was a new day….


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Beauty Box 5 Mani-Pedi Party Bundle – Only $5.00
Disclosure: This post contains affiliate links.
Beauty Box 5 has released a limited edition mani-pedi box. The box is only $5.00, but you will have to pay shipping, unless you spend $50.00.
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What is the phenomenon?
Information about Floods
What are the effects on the landscape and communities?
In floods harvests would be delayed ,crops and pastures would be killed and product destroyed.
Most floods are dangerous and course death.
Houses are destroyed trees are wrecked,shops and buildings are destroyed and wrecked.
How do floods occur?
Where do floods mostly often happen?
Floods occur lots of ways.
These are some of the ways floods can occur,
heavy rain fall,tsunamis,tropical cyclone,
high tide,dam failure triggered and also
high than normal river levels.
Floods also occur when snow melts.
What is a flood?
A flood is water where it is not wanted.Dry land over flows with unwanted water.
There are many places floods occur these are some of the places low lying land,areas adjacent to streams and rivers.
Around the world these are some of the places floods mostly occur, Japan,United States of America,Mexico,China,China's Shanghai River and Taiwan.
What are the safety issues in floods
Lots of people drown in cars or by swimming in rivers or lakes.
Most people think they can swim or drive through floods but they can't make it.
The water looks calm and still but under it's running through faster than you think.
What are the ways of preventing floods
These are some of the ways of preventing a flood building levees,town plan,sand bags in front of the town or in front of your house.
Warning systems for floods
The bureau of Meteorology provides a floods warning service across NSW.
Hello I'm going to explain what floods are. You will understand what floods are after you read this.Enjoy!
Thank you for listening I hope you enjoyed my prezi and learned about floods, remember you can't swim threw floods or drive threw floods.
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Wanna know more about floods click here
cuddles bunnyon 15 July 2014
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text: i'm starting to feel a bit feverish! I have $2,000 to spend on whatever I want..and I want Vuitton! instead of getting 1 or 2 new bags I've decided to head to my lovely MyPoupette resellers and stock up on as many bags as I can get! but there are so many bags to chose from!!!! the only thing I've allowed myself to have is a myrtille blue cles. What I want is 1. BH 2. Speedy 30 3 A petit Noe if anyone sees anything they think that I should see please help! I've had this money to spend for 2 days now and I just can't seem to buy. I'm open to pretty much anythign in good condition. help!:s


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Though partnership has many advantages, one should also see the other side of coin that is disadvantages of partnership. Let's look at some of the disadvantages of partnership –
- One of the biggest disadvantages of partnership is that in partnership partners are jointly and individually liable for the actions of the other partners. So if one of the partners does some mistake, it s will put other partners also in trouble.
- In partnership for taking any decision one has to consult all other partners which delay the decision leading to loss, because in business time is the most important thing.
- In partnership since profit are shared among all the partners it may lead to disputes, if one partner is putting more time into the business and other partners are not putting that much time into the business.
- In case of unlimited liability partnership, partners could lose all of their personal assets in case of bankruptcy of partnership firm.
- A partnership firm comes to an end in the event of death, lunacy or bankruptcy of any partner which in case of established business can be unfair to other partners.
- The amount of capital which partnership firm can raise from outside sources is limited as compared to a listed company and therefore scope of partnership business is limited as no business can survive without fresh capital.
You might also be interested in checking out various advantages of partnership in one of my previous articles.


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Through breathing, movement, and tension reduction, people can also reduce fear and start to focus on enjoying life today.
Greenville, SC (PRWEB) September 02, 2015
Dr. Robert G. Schwartz, the principal of Piedmont Physical Medicine & Rehabilitation, P.A., announced today that he is now certified in "Systema": a Russian martial art system that is also a philosophy of life that focuses on reducing tension in all aspects of life.
Systema, which dates back to the 10th century, is a Russian Martial Art that has four key principles -- breathing, posture, constant motion and relaxation. Each of these principles helps support a system of life that is optimized to reduce tension, and thus can help contribute to the treatment of a variety of health conditions ranging from everyday complaints, to serious diseases. Dr. Schwartz has found the tools and tactics employed by Systema so powerful, that he has integrated them into his medical practice at Piedmont Physical Medicine & Rehabilitation, P.A., which is located in Greenville, SC.
"Through breathing, movement, and tension reduction, people can also reduce fear and start to focus on enjoying life today," commented Dr. Schwartz, who was certified by Vladimir Vasiliev, one of the Systema's founding Directors and Chief Instructors, at its Toronto world headquarters. "Systema helps people to experience the true meaning of freedom. Benefits related to reducing tension include improved endurance, balance, flexibility and self-esteem – all of which can make a significant contribution to any individual's treatment and healing plan, whether they are recovering from illness or want to maintain their optimal sense of health and wellbeing."
Over the last few years Dr. Schwartz has developed specific programs that integrate Systems principles and techniques into his healthcare practice. Updates will be provided on his website at http://piedmontpmr.com. There are already several articles and blog posts on Systema Health.
Dr. Schwartz is also the Systema Instructor at Systema Greenville. Systema classes are held every Sunday at 1:30pm at Cleveland Park in Greenville, SC. More information on Systema Greenville is available at http://www.systemagreenville.com.
For all other information regarding Systema Health including interview requests, contact Sheri Schwartz, Practice Administrator, at (864) 235-1834 or ppmrpublic(at)safe-mail(dot)net.
About Piedmont Physical Medicine & Rehabilitation, P.A.
Robert G. Schwartz, MD has independently owned and operated Piedmont Physical Medicine and Rehabilitation, P.A. in Greenville, South Carolina, USA, since 1984. He specializes in Physical, Vascular and Regenerative Medicine, and offers non-surgical specialty care for complex, chronic pain. Learn more at http://piedmontpmr.com


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We spent one month exploring the sights, sounds, and tastes of Italy. Our cameras were recording from the time we left Prague and continued rolling as we made our way north up through the country. Our favorite stops were the small, quaint hill towns of Assisi, Orvieto, and Civita di Bagnoregio.
Exploring the archaeological ruins of Paestum was an unexpected surprise. Unbeknownst to us, some of the best preserved Greek temples are actually not found in Greece, but rather in southern Italy! Lastly, Venice had a vibe unlike any other city we've ever visited and definitely ranked high on our must see list. We closed out our time in Italy getting lost in it's narrow alleys and taking the budget friendly water bus down the Grand Canal. Before we left this beautiful country, we were granted one last special parting gift as we found 5,000 Russian Rubles floating down the street!
Bonus Clips


text:
Intel® Xeon® Processor D-1533N
- 9 MB Cache
- 6 Cores
- 12 Threads
Intel® Xeon® processor D product family, network series of processors, provides additional advancements at the edge of the network. Supporting delivery of intelligence at the network edge, this product delivers the performance and advanced intelligence of Intel's server-class Xeon® processors in a system-on-a-chip (SOC) form factor, supporting deployments in dense, rugged environments outside of the data center. Network Series processors include up to four integrated Intel® Ethernet 10 GbE adapters and integrated Intel® QuickAssist™ Technology (Intel® QAT), enabling further reduction in the product footprint while also increasing the amount of securely processed and compressed data at the network edge. Infrastructure applications include midrange routers, network appliances, security appliances, wireless base stations, embedded midrange Internet of Things (IoT) devices, entry networking, midrange storage area network (SAN) and network attached storage (NAS) appliances, warm cloud storage, and more.1 2 3
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Enables dynamic provisioning of services as communication service providers extend network functions virtualization (NFV) to the network edge.
Enables smarter and more cost-effective storage solutions through integrated technologies that accelerate data movement, help protect data, and simplify data management.
Support for scalable performance and broad application compatibility.
From multi host controllers to network adapters, Intel® Ethernet products deliver accelerated performance, high bandwidth, and unmatched features for network functions virtualization.
Meet data center efficiency and capacity challenges and the needs of enterprise IT, cloud service providers, and more with intelligent storage solutions powered by Intel® technology.
As Internet of Things (IoT) opportunities grow, Intel is helping connect things to the cloud, integrate with existing infrastructure, and more securely manage data.
Deliver amazing flexibility, scalability, and agility with foundational technology building blocks that come with a virtualized, software-defined infrastructure—the agile, cloud-ready network architectures.
See how open standards–based hardware is creating the cloud-ready network
Hear from Cloudian's CMO, Paul Turner, and ScaleMatrix's cofounder, Chris Orlando, as they highlight how the Intel® Xeon® D-1500 processor family stores and manages unstructured data.
Learn more about managing data in dense data center environments
Watch 1&1 discuss energy consumption and data center space costs, and how the Intel Xeon processor D product family brings improved compute density to their data centers.
See complete speed, performance, and configuration specs.
Intel® technologies' features and benefits depend on system configuration and may require enabled hardware, software or service activation. Performance varies depending on system configuration. No computer system can be absolutely secure. Check with your system manufacturer or retailer or learn more at http://www.intel.co.uk.
Benchmark results were obtained prior to implementation of recent software patches and firmware updates intended to address exploits referred to as "Spectre" and "Meltdown". Implementation of these updates may make these results inapplicable to your device or system.
Software and workloads used in performance tests may have been optimized for performance only on Intel® microprocessors. Performance tests, such as SYSmark* and MobileMark*, are measured using specific computer systems, components, software, operations and functions. Any change to any of those factors may cause the results to vary. You should consult other information and performance tests to assist you in fully evaluating your contemplated purchases, including the performance of that product when combined with other products. For more complete information visit http://www.intel.co.uk/benchmarks.
Cost reduction scenarios described are intended as examples of how a given Intel®-based product, in the specified circumstances and configurations, may affect future costs and provide cost savings. Circumstances will vary. Intel does not guarantee any costs or cost reduction.


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Even if Congress passes the USA Freedom Act, the National Security Agency might continue to collect records on virtually all U.S. phone calls, according to a top Justice Department official.
Senate Judiciary Committee Chairman Patrick Leahy (D-Vt.) has introduced the USA Freedom Act with the stated purpose of ending the NSA's bulk phone record collection program.
Leaks by Edward Snowden showed that the Foreign Intelligence Surveillance Court granted the NSA permission to collect records on all U.S. phone calls under Section 215 of the Patriot Act, even though the law only allows the agency to collect records that are "relevant" to terrorism.
Leahy and other lawmakers have expressed shock that the NSA is using the provision to collect data on millions of Americans not suspected of any wrongdoing.
The USA Freedom Act would amend the Patriot Act to require that the records be relevant to terrorism and that they pertain to an agent of a foreign power or a person in contact with an agent of a foreign power.