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The Glycemic Index: The Real Key To Faster Weight Loss?
(BlackDoctor.org) — Carbohydrates are an important aspect of the daily diet, yet different carbohydrates impact our bodies in various ways. The Glycemic Index (GI) offers a way to measure how various carbohydrates act once they are broken down and enter our bloodstream – which can affect your weight loss goals.
Carbohydrates in the Digestive System
Carbohydrates are mostly made up of sugars, starch and cellulose. When you eat a carbohydrate, it is broken down by the digestive system into glucose, and glucose is one of the energy engines (along with fats and proteins) that keep your body going throughout the day.
Types of Carbohydrates
Carbohydrates come in different forms:

subset: commoncrawl


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"You have to create your own stuff. It's really exciting to create something, sell it, and feel like I'm not just a pawn waiting to be cast."
"One of the things that comedy has given me over the years is a really good ability to laugh at myself and to not take things that don't matter too much too seriously. I feel that very little offends me anymore and I'm really grateful for that because I think I was a pretty uptight little kid."
"My comedy does not come from a place of deep cynicism, and I tend to play characters who are naive in some way."
Anna Faris (born Anna Kay Faris on November 2, 1976 in Baltimore, Maryland) is an American actress and producer. Anna's family moved to Edmonds, Washington when she was six-years-old. Her parents enrolled her into community drama class for kids, which encouraged her to produce her own material in her bedroom with friends. After graduating Edmonds-Woodway High School, she enrolled at University of Washington, earning a degree in English literature in 1999. After graduation, she traveled to London to work for an Ad agency, but pulled out at the last minute to live in Los Angeles. Though she never intended to be an actor, Anna gave it a shot, and she had her breakout role in 2000's Scary Movie. She would reprise her role for three sequels to the horror-parody. Anna started to gain critical praise for her dramatic performances in movies like May, Lost In Translation, Brokeback Mountain and Waiting…, with Ryan Reynolds. In the late 2000s, Anna continued her comedic work, appearing in My Super-Ex Girlfriend, Mama's Boy and The House Bunny. Anna began voice work, lending her talents to animated features like Cloudy With A Chance Of Meatballs and the revival of the Alvin and the Chipmunks franchise. In 2013, Anna was cast in the main role of the CBS sitcom, Mom. It was both a ratings and critical success.
Best Known For:
Anna Faris is best known as an actress, appearing in four of the Scary Movie films and on the television series Mom.
Personal Life:
Anna began dating actor Ben Indra in 1999, shortly after meeting on the indie slasher film, Lovers Lane. They were married in 2004, but Anna filed for divorce in 2007. That year, Anna met actor Chris Pratt at a table read for the film, Take Me Home Tonight. They were engaged in 2008, married in 2009 and welcomed their first son, Jack, in 2012.
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T'is the season
I know we're supposed to be cheerful
when it's Christmas music we hear
playing so endlessly on the radio
but why so early, it's unclear
The festival doesn't frighten me
It's the company I will keep
the holidays can get so lonely
In solitude will I therefore weep
I never cared for relationships
a sign of weakness they are
the pain behind such friendships
leaves behind many a scar
Ti's the season for forgiveness
but for me there's only despair
to gaze helplessly at people happy
for in the past I never cared
Do wishes ever come true?
Will there be hope for me?
Or will I hide way the holidays
Waiting for the new year desperately


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From the Academic Department of Respiratory Medicine, St. Bartholomew's and the Royal London School of Medicine and Dentistry, The London Chest Hospital, London, United Kingdom
1997 by the American College of Chest Physicians
The prevalence of asthma is increasing, despite better understanding of its pathogenesis and improved treatments. During the past 10 years, the perception of asthma has shifted from a disease primarily characterized by altered smooth muscle function to one mainly characterized by chronic inflammation. This article reviews the evidence supporting the relationship of inflammation in both the upper and lower airways, focusing on intermittent seasonal disease as well as on the more chronic and severe forms of asthma, including that associated with aspirin intolerance. It also presents evidence to support a pivotal role for the epithelial cell, together with the mast cell and the eosinophil, in initiating and maintaining inflammation in the upper and lower airways.
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text:
- Service & Staff
- Date & Time
- Your Details
- Confirm
- Finished
Business Information
Tue8:30am - 1:30pm
Wed8:30am - 1:30pm
|(250) 763-9474
Recent reviews
Timely and Effective
Kyle & Megan N - Nov 8, 2016
As we grow into our business as is the need for our relationship to grow to support that business. Beth has been the hidden gem in your corner when you least expect it. Our relationship has grown and therefore so has the business. Forever Grateful!
every. single. time
Jane Howell - Aug 19, 2016
Every single time we meet with Beth - we have massive realizations and solidify 'things'. Sometimes we are not even sure if we need to meet up but then after we are so thankful we did. Its like having an outsider help with bringing clarity - so valuable.
Brilliance and Grace
Justine Armstrong - Aug 10, 2016
Beth is an example of brilliance and grace in action. Also - compassion, wisdom, playfulness, guidance and support. I have benefited so much from my time with her and feel so grateful.
Special Relationship
Cheryl T - May 30, 2016
I took advantage of Beth's free 30 minute session to re-connect after 5 years. Beth remembered me, we were able to catch up quickly and she helped me clarify and 'unstick' myself from a work issue. She's as awesome as ever, even better than before. Everyone needs a coach like Beth!
Real Deal
K.P. - May 11, 2016
Plain and simple, Beth gets down to the nitty gritty! We did exercises that assisted me in getting grounded on my values as a Father and Husband.


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Lifeguard Classes 2017
This 4 day course teaches all the skills for waterfront life guarding. Students must successfully complete the course prerequisites in order to participate. After successful completion of the course participants will be certified in waterfront lifeguarding, CPR/AED for lifeguards, and first aid.
Course Prerequisites:
Must be age 15 by the last day of class.
550 yard swim of front crawl, and breaststroke
20 yard swim, a surface dive to a depth of 10 feet, and retrieval of a 10 pound brick in less than 1 minute 40 seconds.
15 yard underwater swim and retrieval of dive rings.
Tread water for 2 minutes using only the legs.
Course Dates and Times:
Course 1: February 20-23 (Monday-Thursday) CLASS IS FULL
Course 2: April 18-21 (Tuesday-Friday) CLASS IS FULL
Course 3: June 17-18, 24 & 25 (Saturday and Sunday for 2 weeks)
Course 4: June 26-29 (Monday-Thursday)
All classes run 8:00AM-6:00PM
Dress comfortably, bring bathing suit and towels. Pack a lunch and water. If you would like, bring snacks for breaks.
Rates: $300 for Members; $350 for non-members.
A $75.00 non refundable Deposit will hold your registration, balance due on the first day of class.
For more information contact Kat Davis 978-454-7825 x25 or email@example.com


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EGADS 2.0 information and training session
February 8, 2013Tweet
The Graduate School has partnered with ITS to launch a new and improved EGADS (Graduate Admissions Review) experience! The new features of EGADS 2.0 are now live. Join us for an informational session at 1 p.m. today, Friday, Feb. 8, in UUW-206. We will explore the newest features and provide tips for how to get the best use of this valuable tool!


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SINGAPORE: Fuel oil's January viscosity spread, the differential between 180-cst and 380-cst prices, plunged $1.88 a tonne on Wednesday to around a 14-month low of $6.88 according to traders.
BP was seen as the main seller of January viscosity while Hess Corp was actively snapping up offers at around $7.00 a tonne.
Other sellers of the January viscosity contract included Kuo Oil who sold 25,000 tonnes worth at $7.00 a tonne during the Asia trading window.
Similar drops were also seen for the February and March viscosity spreads. The February contract fell $1.63 to $8.88 a tonne while the March contract dropped 25 cents to $10.25.
BP offloaded a total of 90,000 tonnes of January fixed-price 180-cst swaps and bought 25,000 tonnes of January fixed-priced 380-cst swaps during the Asia trading window.
The major also sold a 20,000-tonne physical 180-cst fuel oil cargo for Jan.28-Feb.1 lifting at a discount of $3.75 a tonne to Mercuria, the lowest since Dec. 10, 2012 according to Reuters data.
Fuel oil timespreads were also mostly lower by 1030 GMT with the prompt January/February contract dropped 25 cents to its lowest since Dec. 4, 2012 at a $4.50 a tonne contango.
Physical fundamentals for the Asia fuel oil market has been weak since October 2012 because of lacklustre demand for marine bunker fuel and consecutive months of heavy arbitrage inflows.
Center>Copyright Reuters, 2013


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Thanks for taking action.
Rachel Maddow featured our big event pressuring the White House on Tuesday. And the White House faced questions about the PCCC's advocacy at an official media briefing!
Next, we will run online ads pressuring weak Democrats in Congress to oppose cuts to Social Security benefits.
Can you chip in $3 to ramp up our effective activism and fund these ads? Every $3 helps us reach 3000 people. (Monthly donations help us plan with certainty, so please consider.)
DONATE BELOW or by mail.
You can also donate to the PCCC Strategic Fund to support our big-picture work to change the Democratic Party so things like this don't happen again in the future.
Paid for by ActBlue (actblue.com) and not authorized by any candidate or candidate's committee.
Contributions or gifts to ActBlue are not deductible as charitable contributions for Federal income tax purposes.


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ED Magedson – Founder
Ripoff Report
DriveTime - Ugly Duck11700 US HYW 19 N Clearwater, Florida U.S.A.
Does your business have a bad reputation?
Fix it the right way.
Corporate Advocacy Program™
This report was posted on Ripoff Report on 10/12/2004 11:58 AM and is a permanent record located here: http://www.ripoffreport.com/reports/drivetime-ugly-duck/clearwater-florida-34624/drivetime-aka-ugly-duck-rip-off-clearwater-florida-112573. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
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text:
Eco Art Hotel De Albertis
No prices found through our partners. Please contact the business directly or check some of our recommended alternatives.
How has this hotel rated in the past?
- Multilingual staff
- Continental Breakfast
- Pets allowed
- Family rooms
Photos of Eco Art Hotel De Albertis
Travelers also recommend:
- Pets allowed
- Family rooms
- Multilingual staff
- Continental Breakfast
TripAdvisor Reviews De Albertis Hotel Milan
Travel Blogs from Milan
Visiting a friend in Milano
... lives in Milano. Una visita molto piacevole!
With perfect weather(warmer than usual temperatures) the Piazza del Duomo is filled with people, kids, and pigeons fluttering by. Even with the piazza brimming with people, the Cathedral is center ...
First Day @ Milano
... was "Hotel Bernina"
This is a simple hotel, with simple rooms, with a simple breakfast, that will do job.
If you want a pool, a breakfast with 3 miles and a runaway to put the car...this is not what you are looking for.
to be continued...
Milan - Day 2
... defeat it was all paid for by the Italians, so I guess they had the last laugh.
And what would an Italian Sunday be without church. We didn't attend mass, but we did see the Basilica of St. Ambrose, which was built in the 1080s (although some of it dates back to 379). He is the Patron Saint of Milan and was buried here in 397. To see something of this age still standing and still beautiful is crazy. It makes us hopeful as we get older. Obviously nothing ...
I didn't see any of Milan other than the train station and airport as I was too into my Jack Reacher at night and didn't get to sleep until late. The hostel was brilliant! They will be getting an exceptional review from me! I arrived and had a welcome drink brought to me. The beer was lovely!! On the check in form they asked what your favourite song was. ...
... we pretty much had the place to ourselves and had been on all 20 rides with just over an hour. I was forced onto lots of circular rides which I hate and I nearly lost my dinner on one of them, so opted out for a little while. There were three roller coasters including a wild mouse, a ferris wheel which looked out over the nearby airport which Jesse loved and the boys had a ball. Jesse's They were very excited, because being halloween, they got to stay up past midnight and Jesse ...


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As the vampire snarled at me not to touch her, Jeremy came face to face with her, standing in front of me, death staring her. She left the room. I felt the tears hot on my cheeks as she snarled at me. Jeremy put an arm around me, and we went into the living room. I wiped away my tears. "I'm fine. I just remembered that night. I was wondering if...if he was a vampire. A really well controlled one, by the way he licked his hands after tearing at m mother's throat."
Jeremy put a finger under my chin. "Don't let it get to ya. Tommorows a mystery, yesterday is history, and full of crap, *chuckle* but todays the gift. That's why it's called the present." He told me with a strait face. I gave a laugh. I put my head on Jeremy's shoulder. "Sorry..." I whispered. Jeremy looked down at me.
"I was apoligizing to the vampire, for touching her fangs, to you for causing you trouble, and to myself for being worthless." I whispered. Jeremy fidgeted a bit. He kissed the top of my head. "And I thought Shade needed a chill pill. You need to stop putting yourself down and take a chill pill." He murmured. I smiled up at him, and nuzzled into his shoulder.


text:
I've said before I really like the Smart Business Newsletter that Microsoft produces – there's really useful information in there and I would recommend you take a look if you are a small business and are making IT decisions or want to do smart business using IT.
This month I'm actually part of the action! Check out my article on moving to a client/server architecture here. There are some gems in there that will help you save a lot of money (e.g. leveraging Windows Mobile) whilst maximising your investment (e.g. Active Directory and Group Policy).
I need to have a word with the editors about that picture though 🙂


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Pahlmeyer Napa Valley Proprietary Red 2007
Winemaker Notes
Composition: 85% Cabernet Sauvignon, 7% Merlot, 4%, Cabernet Franc, 3% Petit Verdot, 1% Malbec
Critical Acclaim
The sensational 2007 Proprietary Red Wine (a 7,000-case blend of 82% Cabernet Sauvignon and the rest Merlot, Cabernet Franc, Petit Verdot, and Malbec) is slightly more restrained aromatically than the Merlot, but there is no doubting its extraordinary bouquet of subtle smoked meats, oak, mountain wild flowers, blueberries, blackberries, coffee, and cassis. Super-rich and full-bodied with noticeable, but stunningly sweet tannins, it possesses a rare minerality and definition, characteristics that seem to be found across the board in Napa's 2007s. This dense, powerful red needs 3-5 years of cellaring, and should have 25 years of life ahead of it.
A rich, explosive style, packed with layers of currant, blackberry, wild berry and black cherry fruit woven together in a tight way. Full-bodied, turning supple and polished on the finish, where the fine-grained tannins are ripe and let the fruit flavors flow freely. Cabernet Sauvignon, Merlot, Cabernet Franc, Petit Verdot and Malbec. Drink now through 2020.
Full ruby-red. Slightly syrupy dark fruit aromas complicated by truffle and earth, with an element of minerality providing lift. Then surprisingly ripe and lively in the mouth, with very intense, rich black fruit, licorice and mint flavors. Offers a lovely combination of sweetness and lift and a real impression of small-berry concentration.


text:
Properties, production, and applications of camelid single-domain antibody fragments
- First Online:
- Received:
- Revised:
- Accepted:
DOI: 10.1007/s00253-007-1142-2
- Cite this article as:
- Harmsen, M.M. & De Haard, H.J. Appl Microbiol Biotechnol (2007) 77: 13. doi:10.1007/s00253-007-1142-2
- Citations
- 6k Downloads
Camelids produce functional antibodies devoid of light chains of which the single N-terminal domain is fully capable of antigen binding. These single-domain antibody fragments (VHHs or Nanobodies®) have several advantages for biotechnological applications. They are well expressed in microorganisms and have a high stability and solubility. Furthermore, they are well suited for construction of larger molecules and selection systems such as phage, yeast, or ribosome display. This minireview offers an overview of (1) their properties as compared to conventional antibodies, (2) their production in microorganisms, with a focus on yeasts, and (3) their therapeutic applications.
KeywordsSingle-domain Microbial production Yeast Glycosylation
Although isolated antibody heavy (Utsumi and Karush 1964) and light chains (Yoo et al. 1967) can retain antigen-binding specificity, their affinity and solubility is often reduced (Ward et al. 1989). However, the paired N-terminal variable domains of heavy (VH) and light (VL) chains are sufficient for antigen binding (Sundberg and Mariuzza 2002). Such antibody fragments can be produced as monovalent antibody fragment (Fab) or as single-chain Fv (scFv) where the VH and VL domains are joined by a polypeptide linker (Fig. 1a). Their production in microbial cells is often cumbersome, especially when producing multivalent formats, because of the requirement for domain association.
The discovery that camelids (bactrian camels, dromedaries, and llamas) produce functional antibodies devoid of light chains (Hamers-Casterman et al. 1993) formed a further breakthrough because their single N-terminal domain (VHH, also referred to as Nanobody®) binds antigen without requiring domain pairing. These heavy-chain antibodies also lack the CH1 domain, which in a conventional antibody associates with the light chain and to a lesser degree interacts with the VH domain (Fig. 1b). Although single-domain antibodies later were also identified in particular cartilaginous fish (Greenberg et al. 1995), most research on the biotechnological application of single-domain antibodies was done using camelids because of their ease of handling, including immunization. Methods to isolate antigen-specific VHHs from immune (Arbabi-Ghahroudi et al. 1997; Van der Linden et al. 2000a), nonimmune (Tanha et al. 2002; Yau et al. 2003; Verheesen et al. 2006), or semisynthetic (Goldman et al. 2006) libraries using phage, yeast, or ribosome display are now well established. For further reading on these topics, we refer to recently published reviews (Muyldermans 2001; Dufner et al. 2006).
Sequence analysis (Hamers-Casterman et al. 1993; Muyldermans et al. 1994; Vu et al. 1997; Harmsen et al. 2000) and elucidation of the crystal structure (Desmyter et al. 1996, 2001; Spinelli et al. 1996) has revealed several structural features of VHH domains. Similar to conventional VH domains, VHHs contain four framework regions (FRs) that form the core structure of the immunoglobulin domain and three complementarity-determining regions (CDRs) that are involved in antigen binding. This contrasts with shark single-domain antibodies that have a vestigial CDR2 that does not contribute to antigen binding (Streltsov et al. 2004). As compared to human VH domains, the VHH FRs show a high sequence homology of more than 80%, and their 3D structures can be superimposed (Muyldermans et al. 2001; Holliger and Hudson 2005).
The most characteristic feature of VHHs is the presence of amino acid substitutions at four FR2 positions (positions 37, 44, 45, and 47; Kabat numbering) that are conserved in conventional VH domains and that are involved in forming the hydrophobic interface with VL domains. Occasionally, antigen-binding single-domain antibody fragments that lack these characteristic FR2 substitutions are isolated from camelids. These fall into two groups. The low-affinity binders isolated from a nonimmune library originated from conventional antibodies, presumably because of the polymerase chain reaction crossover cloning artifact, as they were linked to the CH1 domain (Tanha et al. 2002). We refer to these as VHH-like conventional VHs. However, such single-domain antibody fragments with conventional-like FR2 sequences that bind antigen with high affinity are isolated from immune libraries with the high efficiency of about 10% (Conrath et al. 2001a; Saerens et al. 2004; Harmsen et al. 2005a, 2007), which equals their presence in unselected libraries (Harmsen et al. 2000). This is not expected when such clones originate from a cloning artifact. Unlike the clones isolated by Tanha et al. (2002), these clones often contain a hydrophilic residue (mostly arginine) at position 103. This substitution is probably important for their single-domain nature (Desmyter et al. 2001) because conventional antibodies contain a highly conserved hydrophobic residue (tryptophan) at this position that contacts VL. This suggests that these represent functional VHH domains derived from recombination of conventional VH gene segments with heavy-chain constant gene segments during B cell maturation. This was confirmed by the absence of the CH1 domain when such VHH domains were reisolated from the original immune repertoire using a CDR3-specific primer (De Haard, unpublished observation). Therefore, we refer to these as conventional-like VHH domains. Although the increased hydrophilicity of VHHs predominantly relies on the aforementioned changes in the former VL interface, some amino acids at positions that form a slightly hydrophobic patch on conventional VH domains that contacts CH1 (Lesk and Chothia 1988) are also changed into hydrophilic residues in VHHs (Muyldermans et al. 1994; Harmsen et al. 2000).
Furthermore, the CDRs of VHHs contain some characteristic features. Firstly, the N-terminal part of CDR1 is more variable (Vu et al. 1997; Harmsen et al. 2000; Nguyen et al. 2000). Secondly, many dromedary VHHs have an extended CDR3 that is often stabilized by an additional disulfide bond with a cysteine in CDR1 or FR2 (Muyldermans et al. 1994) resulting in the folding of the CDR3 loop across the former VL interface (Desmyter et al. 1996). A particular subfamily of llama VHHs (VHH3) also contains an extended CDR3 that is stabilized by an additional disulfide bond with a cysteine at position 50 in FR2. However, VHHs of this subfamily are rarely isolated, and most llama VHHs have CDR3 loops similar in length to those found in human VHs.
Advantages of camelid single-domain antibody fragments as compared to conventional antibody fragments
Molecular basis
Facile genetic manipulation
Single-domain nature
Increased functional size of immune libraries
No decrease in library size because of reshuffling of VL and VH domains
Facile production of multivalent formats
More flexible linker design and no mispairing of VL and VH domains
Facile production of oligoclonal preparations from single cells
No mispairing of VL and VH domains
High physicochemical stability
Efficient refolding due to increased hydrophilicity and single-domain nature
High solubility
Increased hydrophilicity
Recognition of hidden antigenic sites
Small size and extended flexible CDR3
Rapid tissue penetration, fast clearance
Small size
Well expressed
Efficient folding due to increased hydrophilicity and single-domain nature
Several advantages result from their single domain nature. Thus, VHH libraries generated from immunized camelids retain full functional diversity. This contrasts with the diminished diversity of conventional antibody libraries because of reshuffling of VL and VH domains during library construction. As a result, high-affinity antigen-binding VHHs can be isolated by directly screening a limited number of clones from immune libraries without prior selection using display technologies (Frenken et al. 2000; Harmsen et al. 2005b). Furthermore, the single-domain nature facilitates subsequent molecular manipulation. For example, for many applications, it is advantageous to engineer monovalent antibody fragments into multivalent formats to increase functional affinity (termed avidity) or to produce bispecific antibody fragments that can simultaneously bind to different antigens. Such molecules (diabodies, Fig. 1a) can be produced using conventional recombinant antibodies using linkers between the VH and VL domains of a specific length, although this often results in aggregation and reduced affinity because of mispairing of VH and VL domains (Glockshuber et al. 1990; Whitlow et al. 1993). VHHs are more suitable for production of such formats because they allow more flexible linker design, which is important for simultaneous binding of multivalent antigens, without the problems posed by domain mispairing. Thus, several functional trivalent-bispecific VHHs have been successfully produced (Coppieters et al. 2006; Roovers et al. 2007).
The use of mixtures of a limited number of monoclonal antibodies (oligoclonal antibodies) is advantageous over single monoclonal antibodies for particular applications, such as toxin neutralization (Nowakowski et al. 2002). Because of regulatory requirements, such oligoclonals are preferentially produced from single cells. Again, VHHs are predicted to be more suitable for single-cell production of oligoclonals because of the absence of domain mispairing, although this is yet to be demonstrated experimentally.
Contrary to conventional antibodies, VHHs have been shown to remain functional at 90°C (Van der Linden et al. 1999) or after incubation at high temperatures (Van der Linden et al. 1999; Perez et al. 2001). This high apparent stability is mainly attributed to their efficient refolding after chemical or thermal denaturation and to a lesser extent because of an increased resistance against denaturation (Perez et al. 2001; Dumoulin et al. 2002; Ewert et al. 2002). The increased apparent stability is probably due to an increased hydrophilicity of the former VL interface region because a "camelized" human VH fragment that contains several of the hallmark hydrophilic amino acid residues of VHHs was more stable than the original VH fragment (Davies and Riechmann 1995, 1996), whereas "decamelization" of a VHH to mimic a VH domain reduces its thermodynamic stability (Conrath et al. 2005). In addition to these specific mutations, the packing of extended CDR3 loops against this former VL interface contributes to domain stability (Bond et al. 2003). Furthermore, refolding of VHHs only requires domain refolding, whereas conventional antibodies also require association of VH and VL domains.
VHHs can also recognize antigenic sites that are normally not recognized by conventional antibodies such as enzyme active sites (Lauwereys et al. 1998; De Genst et al. 2006) and conserved cryptic epitopes (Stijlemans et al. 2004). This facilitates their use as enzyme inhibitors or in diagnosis of trypanosome infections. The ability to recognize these recessed antigenic sites has been attributed to their smaller size and the ability of the extended CDR3 loop to penetrate into such sites (Desmyter et al. 1996; De Genst et al. 2006). It is interesting to note that this structure–function relation is also observed in a rare example of a broadly reactive human mAb that recognizes the recessed and conserved CD4-binding cavity of human immunodeficiency virus type 1 gp120 by virtue of an extended CDR3 (Zwick et al. 2003). With respect to antigen binding, the single-domain nature could be a disadvantage for binding to small antigens such as haptens and peptides because these normally bind in a groove or cavity at the VH–VL interface (Sundberg and Mariuzza 2002). Indeed, llamas immunized with clenbuterol developed conventional but not heavy-chain antibodies against this hapten (Lange et al. 2001). However, hapten- and peptide-binding VHHs have been successfully isolated using strong selection systems (Spinelli et al. 2000; Yau et al. 2003; Alvarez-Rueda et al. 2007; Harmsen et al. 2007). The affinities of VHHs are generally comparable to those of conventional antibody fragments (Muyldermans et al. 2001). Occasionally, VHHs with affinity constants (KD) as low as 100 pM are isolated (Saerens et al. 2004; De Genst et al. 2006; Harmsen et al. 2006), which equals the affinity ceiling proposed for natural antibodies (Sundberg and Mariuzza 2002).
Because of their small size of about 15 kDa, VHHs rapidly pass the renal filter, which has a cutoff of about 60 kDa, resulting in their rapid blood clearance. In addition, the small size results in a fast tissue penetration. This is advantageous for targeting of VHHs coupled to toxic substances to tumors (Cortez-Retamozo et al. 2004), in vivo diagnosis using imaging, and treatment of snake bites (Harrison et al. 2006). However, for other therapeutic applications, such as treatment of infectious or inflammatory diseases, the short serum half-life of about 2 h (Cortez-Retamozo et al. 2002; Harmsen et al. 2005a) is a disadvantage.
Production in microorganisms
Although a fully active nonglycosylated IgG was recently produced at high level in Escherichia coli, most functional complete antibodies can only be efficiently produced using mammalian cells, especially when their appropriate glycosylation is required for therapeutic applications. However, antibody fragments that lack the Fc with its N-linked oligosaccharide are preferably produced in microbial systems (Arbabi-Ghahroudi et al. 2005). These have a shorter development time from gene to product and require simple well-established fermentation conditions that can be performed on large-scale resulting in costs of goods that can be as low as $1 per gram heterologous protein (Estell 2006). Most large-scale microbial production systems are based on E. coli, yeasts, or filamentous fungi. Production in E. coli can be done by secretion into the oxidizing periplasmic space or expression in the reducing cytosol. The latter requires the often cumbersome refolding of antibody fragments (Arbabi-Ghahroudi et al. 2005). Using eukaryotic microbial hosts, antibody fragments are generally produced by targeting to the secretory pathway. This enables efficient disulfide bond formation, addition of N-linked oligosaccharide, and secretion of soluble, correctly folded product to the culture medium.
VHHs have often been produced in E. coli (Arbabi-Ghahroudi et al. 1997; Rahbarizadeh et al. 2005). There is only one example of VHH production in filamentous fungi, which resulted in limited proteolytic degradation of the secreted product (Joosten et al. 2005) because of the high levels of proteases secreted by filamentous fungi (Gerngross 2004). VHHs have also often been produced in the yeast Saccharomyces cerevisiae (Frenken et al. 2000; Thomassen et al. 2002; Van der Vaart 2002). VHH production by the favored yeast expression host Pichia pastoris was only recently described (Rahbarizadeh et al. 2006). Occasionally, yeast-produced VHHs are N-glycosylated (Frenken et al. 2000; Harmsen et al. 2005a). This can affect antigen binding (Van der Vaart et al. 2006). Furthermore, it could complicate their therapeutic use because the addition of yeast-specific high-mannose oligosaccharides results in a high immunogenicity and decreased serum half-life because of binding to specific mannose receptors on cells of the reticulo-endothelial system (Sethuraman and Stadheim 2006).
Although VHHs are generally well produced in microorganisms, the production level of different clones can vary by a factor of 100 (Frenken et al. 2000; Harmsen et al. 2005b; Van de Laar et al. 2007). Several VHH sequence patterns can be associated with their production level. First, the presence of a potential N-linked glycosylation site often increases production levels in yeast (Sagt et al. 2000). Second, in our experience (Harmsen, unpublished observations), conventional-like VHHs are generally produced at reduced levels in yeast. This contrasts with the reported efficient production in E. coli of VHH-like VHs (Tanha et al. 2002) but is consistent with the increased production level of "camelized" conventional VH domains in E. coli (Davies and Riechmann 1995). Third, the presence of unpaired C-terminal cysteines reduces expression levels (Simmons et al. 2006). Fourth, replacement of hydrophobic residues of conventional VH domains normally interacting with CH1 increased scFv production in E. coli (Nieba et al. 1997), suggesting that the hydrophilic mutations that naturally occur at these positions in VHHs also contributes to their high expression level. However, there are many examples of VHHs that differ by only a few amino acids and are produced at highly variable levels where the exact amino acid change responsible for the difference in production level is difficult to predict (Frenken et al. 2000; Harmsen et al. 2005b). Furthermore, without such knowledge, VHH production can be improved by random molecular evolution using deoxyribonucleic acid shuffling (Van der Linden et al. 2000b), as has often been done for conventional antibody fragments (Dufner et al. 2006). The high refolding capability of VHHs, which is a consequence of their sequence, has also been correlated with a high production level in E. coli (Jespers et al. 2004; Olichon et al. 2007).
In addition to the nature of the VHH, host factors affecting VHH production have been identified. In baker's yeast, the specific VHH production rate is correlated with growth rate (Thomassen et al. 2005) and can be up to fivefold increased by growing on ethanol as the carbon source (Van de Laar et al. 2007). Supplementation of the medium with sorbitol, casamino acids, or ethylenediamine tetraacetic acid improves VHH production by P. pastoris (Rahbarizadeh et al. 2006).
In addition to monovalent VHHs, several expression formats for the production of VHH multimers have been described (Fig. 1b). These include genetic fusions of two (Conrath et al. 2001b; Harmsen et al. 2005a) or three VHHs (Coppieters et al. 2006; Roovers et al. 2007) that either recognize different antigens or the same repeating antigen to increase functional affinity. Although such VHH fusions are less efficiently produced than their monovalent versions, their production level exceeds that of their conventional-antibody-based fusion counterparts without aggregation or low solubility. However, antigen binding by the C-terminal VHH in such fusions can be compromised (Conrath et al. 2001b) presumably because of steric hindrance by the N-terminal VHH. The avidity of VHHs has also been strongly increased using genetic fusions to the B-subunits of an E. coli toxin that self-assembles into a homopentamer (Zhang et al. 2004), resulting in pentameric recombinant antibodies ("pentabodies," Fig. 1b).
VHHs on their own cannot recruit effector functions such as ADCC and CDC. This limits their therapeutic application. Although such effector functions can be indirectly recruited using bispecific (conventional) antibody fragments binding to host immunoglobulin (Holliger et al. 1997), it may be more efficient to recruit these functions by fusing VHHs to host Fc domains. Production of such functional antibodies requires the correct glycosylation of the CH2 domain, which until recently could only be accomplished using higher eukaryotic cells (Nguyen et al. 2003) but not by microbial production. However, this may now be feasible using P. pastoris strains with an engineered glycosylation machinery that are able to produce proteins with a specific human glycoform (Hamilton et al. 2006). Furthermore, transgenic mice containing hybrid llama/human antibody loci that contain llama V regions and human D, J, and C regions have recently been used to generate human heavy-chain antibodies in mice (Janssens et al. 2006).
Therapeutic applications
Examples of therapeutic applications of camelid VHHs
Target antigen
VHH valency for disease target
Additional fusion partner
Sleeping sickness
VSG oligomannose
Apolipoprotein L-I
Baral et al. 2006
Infant diarrhea
Van der Vaart et al. 2006
Infant diarrhea
Lactobacillus cell-surface anchor
Pant et al. 2006
Piglet diarrhea
E. coli
F4 fimbriae
Harmsen et al. 2006
S. mutans
I/II adhesion
Kruger et al. 2006
FMD virus
Harmsen et al. 2007
N. meningitidis
El Khattabi et al. 2006
Cortez-Retamozo et al. 2004
EGF receptor
Anti-albumin VHH
Roovers et al. 2007
Rheumatoid arthritis
Anti-albumin VHH
Coppieters et al. 2006
Brain disorders
α (2,3)-Sialoglycoprotein
Muruganandam et al. 2002
Neurodegenerative diseases
Gueorguieva et al. 2006
VHHs are especially suited for oral immunotherapy because of their resistance against extremes of pH and the capacity to bind to the target at high concentrations of chaotropic agents (Dumoulin et al. 2002, 2003). Administration to piglets of a VHH that effectively prevents intestinal attachment of E. coli bacteria that cause diarrhea resulted in poor in vivo protection (Harmsen et al. 2005b) because of degradation by gastrointestinal proteases (Harmsen et al. 2006). However, by selection for proteolytic stability, a VHH could be isolated from the original library that was not degraded in vivo (Harmsen et al. 2006). VHHs that successfully prevented diarrhea caused by rotavirus in a mouse model were similarly selected for resistance against the acidic environment of the stomach (Van der Vaart et al. 2006). Alternatively, VHH proteolysis can be prevented by local VHH production using natural gut commensal bacteria. Thus, diarrhea could also be prevented by lactobacilli that produce rotavirus-neutralizing VHHs fused to a cell surface anchor (Pant et al. 2006). Treatment of caries, caused by Streptococcus mutans, with VHHs conferred only limited protection (Kruger et al. 2006). Because these VHHs should function in the oral cavity, the low level of protection cannot be due to proteolytic VHH degradation within the gastrointestinal tract.
The short serum half-life because of a rapid renal clearance limits the efficacy of VHHs in many parenteral applications. Therefore, VHHs have been targeted to normally long-lived serum proteins such as albumin (Coppieters et al. 2006; Roovers et al. 2007) or immunoglobulin (Harmsen et al. 2005a) using bispecific VHHs recognizing these serum proteins in addition to the therapeutic target, resulting in half-lives that equal the half-life of albumin (2 days in mice) and immunoglobulin (9 days). An alternative well-known approach to increase serum half-life of proteins is the chemical addition of polyethylene glycol (PEG). Such PEGylation of foot-and-mouth disease (FMD) virus-neutralizing VHHs not only increased serum half-life but also increased in vitro neutralizing potency to levels above that of the hyperimmune serum (Harmsen et al. 2007). However, in contrast to the full protection afforded by the hyperimmune serum, these VHHs poorly protected guinea pigs from FMD viral challenge infection, suggesting that Fc-mediated effector functions are required for efficient in vivo protection (Harmsen et al. 2007).
Nevertheless, many diseases were successfully treated with VHHs in the absence of Fc-mediated effector functions. These VHHs either are used as targeting devices for toxic enzymes or block a specific molecular interaction. For example, sleeping sickness was successfully treated with VHHs that bind to a trypanosome coat protein and were fused to the apolipoprotein L-1 enzyme, resulting in trypanosome lysis (Baral et al. 2006). In oncology, a VHH directed against carcinoembryonic antigen was used for targeting the genetically fused β-lactamase to tumor cells. This enzyme then converts an injected nontoxic prodrug into a toxic drug in the vicinity of the targeted tumor cells, leading to their killing (Cortez-Retamozo et al. 2004). Several VHH therapies are also being developed for treatment of oncology or inflammatory diseases based on blocking molecular interactions. VHHs binding to epidermal growth factor receptor (EGFR) can block epidermal growth factor (EGF) binding to its receptor, which can be used to treat solid tumors (Roovers et al. 2007). Tenfold more potent EGFR-binding VHHs could be obtained by construction of bivalent formats. It is interesting to note that the recently approved conventional antibody Panitumumab directed against EGFR also blocks EGF binding and is expected to give poor ADCC and CDC (Reichert and Valge-Archer 2007). Furthermore, by blocking receptor interaction, VHHs binding to tumor necrosis factor-α can be used for treatment of rheumatoid arthritis (Coppieters et al. 2006). The potency of bivalent formats was 500-fold increased as compared to monovalent VHHs and even exceeded the potency of clinically used conventional antibodies both in vitro and in a murine arthritis model. Similarly, lipopolysaccharide (LPS)-binding VHHs were isolated that block LPS binding and signaling to host cells for treatment of sepsis (El Khattabi et al. 2006).
The potential immunogenicity of VHHs could compromise their parenteral therapeutic use, especially in treatments that require repeated injections. Until now, multiple injections of VHHs have not shown any immunogenicity in mice, as assessed by the presence of specific antibodies, T cell proliferation, or cytokine levels (Cortez-Retamozo et al. 2002; Coppieters et al. 2006). This could rely on their high sequence homology to conventional VH domains and on their high stability because aggregation of proteins is known to increase immunogenicity (Hermeling et al. 2004). If necessary, technologies developed to decrease immunogenicity of mouse monoclonal antibodies (Presta 2006) could also be applied to VHHs. Alternatively, immunogenicity could be reduced by the use of conventional-like VHHs, which have an even higher structural homology to conventional VH domains.
For their use in targeting drugs across the blood–brain barrier (BBB) into the brain, VHHs were selected that transmigrate the human BBB in an in vitro model and accumulate in the brain after intravenous injection into mice (Muruganandam et al. 2002). These could be used for treatment of neurological disorders. Finally, Bax-specific VHHs have been expressed in the cytoplasm, resulting in so-called intrabodies, to prevent oxidative-stress-induced apoptosis that is implicated in several neurodegenerative diseases (Gueorguieva et al. 2006). Because of their stability, VHHs are especially suited for intrabody production because this requires expression in the reducing environment of the cytoplasm (Gueorguieva et al. 2006; Rothbauer et al. 2006).
Since the discovery of heavy-chain antibodies in 1993, the field of single-domain antibody fragments has been rapidly growing. VHHs have many advantages for biotechnological applications. They can be economically produced in microorganisms and have a high stability. Furthermore, they are highly suited for expression as multivalent, including bispecific, formats or as enzyme fusions. This permits a plug-and-play approach, where, depending on the target, biology potency can be increased by multivalent constructs or bispecific VHH recognizing two different targets can be made. This also enables the tailor-made design of serum half-life using site-directed PEGylation or by targeting to long-lived serum proteins using bispecific VHHs. Although fusions of targeting VHHs to Ig-binding VHHs or Fc can be used to recruit effector functions most current research on VHHs focuses on therapeutic applications where such effector functions are not required. Finally, conventional whole antibodies occasionally give side effects because of their bivalent nature, which can result in target cross-linking, or the presence of the Fc region. Evidently, such side effects are not expected to occur using monovalent VHHs. This, however, is yet to be confirmed as the first VHH has entered phase I clinical trials in 2007 (http://www.ablynx.com).


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Hip-hop veterans Naughty by Nature - accompanied by Sway & The Wonder TwinZ - recently completed a USO tour to the Persian Gulf and what a trip it was!
This marked the second tour for trio, who entertained and met with more than 1,500 troops serving in Kuwait and Iraq in 2009. This was the first USO tour for MTV correspondent Sway Calloway, who served as emcee.
Vin Rock had this to say before leaving, "I'm excited to support our troops again by touring with the USO. We enjoyed ourselves the previous trip, learned alot and were happy we could put smiles on the troop's faces." And judging from Troops' responses, the concerts were a huge hit.
Check out video footage from their first tour stop at Camp Lemonier!
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More from the USO
May 25, 2017
NBC's 'This Is Us' Cast Visits Service Members and Military Families on USO Tour at Joint Base McGuire-Dix-Lakehurst
It was a morning to remember. A group of stars from NBC's "This Is Us" – Mandy Moore, Milo Ventimiglia, Chrissy Metz, Justin Hartley, Susan Kelechi Watson and Chris Sullivan – spent a morning visiting service members and their families at Joint Base McGuire-Dix-Lakehurst as part of a USO entertainment tour.
May 24, 2017
After Struggling to Find His Place in the World, Marine Finds Two Families – and His Place in Both
Deron Johnson didn't plan on joining the military, but he ended up a Marine. He also didn't anticipate becoming a fraternity brother at Texas A&M, but then came Theta Chi and its history of supporting the military - and the USO.
May 15, 2017
USO Operation Birthday Cake Brings the Party to Service Members in Japan
When Navy Petty Officer 3rd Class Jeffrey Lee received an urgent call to head to his work station one morning, he thought there was something seriously wrong, but there was no emergency. Instead, he walked into his own surprise birthday celebration, courtesy of his wife, USO Yokosuka and the Operation Birthday Cake Program.


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Bright and comfortable appartment in central Paris, close to everything : shops, museums, theaters, city highlites. In a quiet district and friendly neighborough , you have lots of shops and restaurants at the bottom of the appartment and a little open market twice a week.
To organize a home exchange, you need to be a member


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George Osborne must answer claims that he placed undue pressure on the Co-operative Bank to buy 632 Lloyds branches, according to the chair of the parliamentary Treasury committee.
Andrew Tyrie has written to the chancellor demanding that he addresses allegations of political interference made by Paul Flowers, the Bank's former chairman.
Flowers, who resigned and was later filmed allegedly buying drugs, claimed this week that politicians put intense pressure on the bank to go ahead with an ill-fated bid for the branches, then known as Project Verde.
In a letter published on Thursday, Tyrie asked Osborne: "Did Treasury ministers or any Treasury officials at any time bring undue pressure to bear on the Co-operative Bank or Group, or Lloyds Banking Group, in respect of the sale of the Verde branches?
"The committee would be grateful if you could confirm what contacts you, other Treasury ministers (in particular Mark Hoban and Greg Clark) and Treasury officials had in respect of the Verde bidding process."
He said the committee would question Osborne on the claims when the chancellor appears to answer questions on the budget next Thursday.
Flowers claimed the bank received calls from then junior Treasury minister Hoban two or three times a week.
"There was pressure certainly from Mark Hoban, but I believe and know that originated much higher up, with the chancellor himself," Flowers told BBC Newsnight on Tuesday.
Flowers said Hoban had "many, many" telephone conversations and meetings with Peter Marks, the then chief executive of the Co-op Group.
Hoban has denied putting pressure on Co-op. "When you go through the Treasury records as we have done, I think I had two meetings with the Co-op, and 10 phone calls," he said last year. Hoban said none of his conversations were with Flowers, and most of the meetings were after Co-op had signed up as bidders.
Marks told the Treasury committee last year that he was not put under any political pressure.
The sale of the Lloyds branches was required by the European Union as a condition of Lloyds' taxpayer-funded bailout
In Flowers's now infamous appearance before the Treasury committee last year, he said he had regular conversations with ministers but said he was not put under pressure to bid for the branches.
"I would not say [it was] pressure. It was mainly a sense of political goodwill behind the process but there were plenty of politicians who wanted the deal to succeed and were willing it on," he said.
This week former Bank of England governor Lord King denied a separate claim of political interference in the deal. Lord Levene, the former chairman of NBNK, which tabled a rival bid for the branches, claimed King told him NBNK's bid would fail because a "political decision" had been made to support the Co-op bid.
King, who retired as Bank of England governor in June 2013, produced notes taken during meetings with Levene to demonstrate that he had not suggested politicians were wedded to the Co-op bid. However, the former governor admitted he was concerned about repeated delays to the sale of the branches.
A Treasury spokesman said: "The decision on whether to proceed with the Verde deal was a commercial matter for Lloyds Bank and the Co-op Bank, as the board of Lloyds has made consistently clear. Since the full extent of the situation at Co-op Bank became clear the chancellor has ordered an independent investigation into the events at the Co-op Bank and the circumstances surrounding them."
The Co-op pulled out of the bid last year and within two months revealed a £1.5bn black hole in its accounts. The Co-op was bailed out by bondholders – led by US hedge funds – who now own 70% of the bank. This week the Co-op admitted it needs to find an extra £400m to pay compensation for mis-selling to customers.
Following the collapse of the sale to the Co-op, Lloyds decided to float the branches on the stock market as a standalone bank - TSB Bank - this summer.


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In the village of Ždiar, within 1.9 miles from the ski centres of Strednica Zdiar and Bachledova, Vila Artemis offers a fully equipped shared kitchen and a common living room with a fireplace and a TV. Wi-Fi is available for free. Offering a balcony and views of the surrounding Spišská Magura hills and Belianske Tatry mountains, all rooms come with satellite TV and fridge. Each unit also comprises a private bathroom with a shower. Continental breakfast is served each morning. Guests can also relax in the garden with barbecue facilities. Other on-site facilities at the Artemis Vila include ski storage and private parking. The ski centre of Tatranska Lomnica is 10.6 miles away, while the Belianska Cave is reachable in 4.3 miles. The cycling track of Tatranska Kotlina - Spisska Bela passes 1,313 feet from the Artemis.


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Creation's Careful ConfinesMature
It was a welcomed escape from the light that struck her and the tilt of the wagon that churned her body. Now, she knew herself to be somehow elsewhere.
The landscape was familiar. The empty water and the uninhabited forest met at a stony shoreline. She was above and within it all, and knew herself as more than anything bound to physical limbs and nerves.
She relaxed, stretching deep into the roots and high into the clouds. Never straying from her peninsula of land, she also made sure to keep away from the nothingness that frightened her.
It was immobile, and posed no threat.
Modesty found herself to be tranquil. And in that serenity, like a sleeping child, she rolled over. No, not in any physical way, but in a way that pressed against that edge.
Her awareness rubbed against the edge. It rippled with plasmatic tension, and then gave way like broken hymen. However, it bled outwards and not inwards. Giving itself away, it opened up to a more tangible nothingness.
To Modesty, it felt as if a door had opened. And at this point, she found herself to be alone no longer. Behind her, down the peninsula, came a second awareness.
Where she bore an aura of soft and gentle green, she saw that he was a wisp of dark red. It was a colour she'd seen before. His existence was like a scarf circling in the ethereal wind. And so, she knew him as she'd seen that scarf before.
She felt drawn to him.
"Do not be so bold, my dear lady," he communed.
Backing away, she pushed further against the nothingness until it gave way to bare canvas of earth. Taking shape beneath her feet, it extended from the forest's edge in nervous crags of wasteland, rising in a hill like a shelter from him.
"Please, do not stretch what does not need yet be stretched. Plenty is bare as is. And your forest, it lacks life and spirit."
Confused, she pushed questions back at him, "My forest? Oh, of course. This is a dream. All a figment of my imagination, to be sparsely remembered. Everything I see and do is mine, including you."
The red wisp smiled, returning, "Oh, I wish I was yours. Like that moment years past, my harp and your neck."
"This is no manner of dream as you know it, Modesty."
"Proof it," she communed, "if you can."
"I'll let you do the proving. Try and wake up. Like a nightmare, where a beast has chased you into a blocked off alley. Will yourself not to me, but to wake up."
Her awareness winced and trembled with effort, managing only to parch the earth around her. Again, she tried, and he watched on as a hemlock withered.
"You have much to learn," he said.
"Uhmm... well, you see, we have no blocked alleyways in Wahleiss."
"Don't give me your excuses. You have dead-end horse stalls at least."
She recoiled in frustration, blowing wind through the trees.
"I have been obliged to teach you our craft," the bard said, as he whirled into the form of himself. He spoke looking up at the trees. "Partly because I passed it to you, and partly because of forces that seek to control you."
A voice came back to him from the trees, a dull and monotone statement, "I have been kidnapped by a troupe of entertainers."
"What? By the Goddess, so soon? I- I must... I must go."
And in an instant, the Bard of Silver Lips vanished, leaving a shiny set of lips to fade away against the dirt.
Modesty willed herself to take form, and felt the contortions of awareness and energy take shape. She descended upon the forest's edge as a very large gray rabbit, and hopped forward into the emerging wasteland.
Unable to wake herself, despite further attempts and having seen the nameless harpist leave, she decided to make herself at home. First, she drove away the nothingness that fringed her piece of nowhere. Then, she let the sea meet its edge with a sandy beach line. But here and there, emerged the gray rocks from the forest.
Turning around, she realized that the wasteland met the forest's edge in a strikingly unrealistic manner. It was straight-edged like a knife. So, she concentrated on adding a seemingly random edge of more trees, which descended into meadows and the occasional glade of remaining woods. And finally in the distance, stretched her cold hard wasteland of resentment.
And all of it was devoid of life, other than the vegetation her mind had conjured. Carefully, she focused, and a fish dropped onto the shore. It was long, blue, shiny and sharp. It wiggled about in the air. Quickly, she manipulated it such that it leaped gracefully into the water.
Then, came more. Fish fell into the water. Snakes slithered across the barren plains. Tiny rabbits dug burrows in the pine forest. A bear emerged, looking for a mate.
Modesty realized that, now, she had disobeyed her supposed instructor. Now, there was new land which she had shorn from the nothing. She couldn't help but wonder what that might cause. And wonder as she might, she realized how truly alone she was.
But try as she might, Modesty could not bring about anything more sentient than a dolphin. And as the hours passed by, she soon woke to a nauseating smell.


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|"Caldarale, Charles R" <Chuck.Caldar...@unisys.com>
|RE: confusion of tomcat downloading
|Wed, 30 Nov 2005 15:18:12 GMT
> From: blueberry lake [mailto:firstname.lastname@example.com] > Subject: Re: confusion of tomcat downloading > > I downloaded tomcat for unix, and want it to run inside apache. To repeat: Tomcat does not run _inside_ Apache httpd, but it can be configured to work with httpd as a front-end using the AJP13 connector. But unless you're doing load balancing or some such, why do you think you want httpd? - Chuck THIS COMMUNICATION MAY CONTAIN CONFIDENTIAL AND/OR OTHERWISE PROPRIETARY MATERIAL and is thus for use only by the intended recipient. If you received this in error, please contact the sender and delete the e-mail and its attachments from all computers. --------------------------------------------------------------------- To unsubscribe, e-mail: email@example.com For additional commands, e-mail: firstname.lastname@example.com
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Posted May 27, 2013 by Kris-E in Movies & TV
Ludacris Admits that he Still Drives his '93 Acura! [Video]
TV talk show host Chelsea Handler, brought Ludacris on her late night show Chelsea Lately. Luda admits to Chelsea that he still drives the same '93 Acura that he rode around in before he became famous. The two continue to talk charity and cars and how Chelsea drives a Bentley, after doing the do with a rapper (which was reportedly 50 Cent.)
Some things we just can't let go!


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Rs. 18,900.00 Is our price too high?
The DragonFly is a very popular USB stick sized DAC+Preamp+Headphone Amp by AudioQuest that was launched in 2012. Till date, the DragonFly counts multiple awards to its name: Stereophile's 2012 "Computer Audio Component of the Year", 2012 "Budget Component of the Year;" Tone Audio's 2012 "Digital Product of the Year;" Computer Audiophile's 2012 "Computer Audiophile Product of the Year;" AudioStream's "Greatest Bits;" and What Hi-Fi?'s 2014 "Product of the Year.
AudioQuest defined the market for premium micro-DACs with the original DragonFly and audiophiles everywhere loved this small product for its great audio quality. However, they weren't capable of being used with any other device apart from the PC.
The big difference between the DragonFly Red and the DragonFly Black is that the Red can drive even the most power hungry high impedance headphones with effortless ease. Whether you're listening to Spotify, YouTube, Tidal, ripped CDs, or state-of-the-art high-resolution files, DragonFly will deliver a more emotionally compelling and enjoyable experience.
The new DragonFly Red is now more portable, and can be used with Android and Apple smartphones in addition to being used with a PC. Now music listeners can enjoy the convenience of a portable DAC that they can carry in their pockets everywhere they go.
Made in Collaboration with Gordon Rankin & Microchip TechnologiesThe DragonFly Red has been developed by AudioQuest in partnership with designers Gordon Rankin and Microchip Technologies. The new DragonFly has a better signal to noise ratio and much lower power consumption than the original version. How much lower consumption you ask? 77%! This is thanks to the new Microchip PIC32MX microcontroller that enables true compatibility with Apple and Android smartphones and tablets.
Asynchronous USB Streamlength®Streamlength™ asynchronous USB code ensures low jitter, low resource load, minimal packet errors, world-class audio playback, and reliable connectivity between our DAC and any computing device compliant with USB Host Mode (as set forth by the USB Organization). Streamlength requires no additional drivers, making DragonFly Black virtually plug-and-play for Apple, Windows, iOS, and Android users.
Meanwhile, with its higher 2.1-volt output, DragonFly Red will be compatible with a wider range of headphones, including power-hungry, low-efficiency models. Its processing capabilities to just 24-bit/96kHz resolution. This makes using the DragonFlys as simple as it's always been: they're fully compatible with PCs without having to download and install new drivers.
Upgradeable Software through Free Desktop AppThe DragonFlys are software upgradeable thanks to the free Windows or Mac desktop app. As and when there are any updates available to music player apps, streaming protocols, or other software, the complimentary app will let you incorporate these updates to any existing devices.
Bit-Perfect Digital Volume Control
DragonFly Red employs a 64-bit, bit-perfect digital volume control that resides inside the DAC chip itself—an elegant and sophisticated implementation that ensures maximum fidelity, dynamic contrast, and signal-to-noise ratio. With DragonFly Red connected to a PC or mobile device, adjusting the host's system volume control will, through proxy, control the DragonFly Red's onboard volume.
What's in the box:


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Dolly Parton and Miley Cyrus join Pentatonix for 'Jolene' on The Voice
Parton and Cyrus had previously performed the song together at Dollywood's 25th anniversary
Parton and Cyrus had previously performed the song together at Dollywood's 25th anniversary
It was a chance encounter in a Nashville parking lot that did it.
Alice Jones' Arts Diary
US officials have said it is too soon to determine whether prescription drugs were involved in Whitney Houston's death.
The 'Wire in the Blood' killer is back, but his creator Val McDermid tells Danuta Kean there is more to her work than violent crime
Designers are being inspired by music stars this season – Miu Miu's latest heroine certainly knows how to rock a look, says Susannah Frankel
A more-than-defensible collection of Jones's collaborations over the past decade: 18 songs in the company of luminaries as diverse as Ray Charles and Belle & Sebastian, Willie Nelson and OutKast, Dolly Parton and Herbie Hancock.
Asked by: Stephen Ward, Poole
Answered by: Dr John Powell, a Visiting Professor of Materials Science at the University of Nottingham
A little bit country, a little bit rock'n'roll, a hint of Dolly Parton and an avalanche of Carlene Carter.
Work: Presents 'The No Nonsense Breakfast Show With Ian Camfield' on Xfm London 104.9 FM, weekdays from 7am-11am
Life: Aged 31, he lives in Belsize Park, north London
Balance: Going to gigs (Spinal Tap, Steely Dan and Blur in the past week). I'm trying to develop a "gym hobby"
Singer, businesswoman, philanthropist – and now Broadway impresario. It's time to claim the country star as a feminist icon, argues Joan Smith
Socialite Tamara Beckwith used to beg Bob Carlos Clarke to take photos at her parties. Now she's setting up a gallery in his memory.
Emmylou's Daniel Lanois period is over. She's back in Nashville with Brian Ahern on the other side of the thickened glass, which means that an elegantly chiming country-rock sound replaces the arty false perspectives and cultivated grit of the Lanois vibe.


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Welcome to BUSC! BUSC (the British Universities Snowsports Council) is the student snowsports governing body responsible for creating key Ski and Snowboard events for over 70 universities and institutions, across the student snowsports calendar. We are all about snowsports and want you to come and party with us!
We are unique in the way we are the only organisation sanctioned by BUCS (British Universities and College Sports) allowing students to win points for their university in all the competitions. But don't think that the excitement ends when the sun goes down, we are striving to bring some of the biggest nights out you've ever seen in a ski resort! Pushing the resorts to their limits to give you the most memorable ski holidays of your lives.
We want to see as many new faces at our events as possible, and are passionate about grass roots competitions. For over 20 years BUSC has been helping the student snowsports community to thrive, providing a platform for competitive events and huge trips with amazing production.
We are run by students for students, and have a passion for events that go above and beyond what is expected. Come and share our passion with us. Please take a look around the website at the variety of events you can get involved in this season!!


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THE ISLE of Man Law Society has voted to allow multi-disciplinary and multi- national partnerships on the island.
A special working party has now been set up by the society to examine the best way of introducing MDP and MNP laws after proposals to use them were voted through at a special meeting of the council.
At present only a handful of international law firms have set up shop on the island and there are only 40 English qualified solicitors based there.
The society president, Geoffrey Karran, said the vote had been prompted by the need to attract the best international commercial law-yers to the island. he added: "Our own bar has developed considerably, but we are are looking to find ways of improving the service we can offer our clients and attracting business to the island."
He said the working party, which is due to report next March, had already received enquiries from accountancy firms interested in the developments.
Jenifer Stanley, a solicitor at Travers Smith Braithwaite, the only City firm with an office on the island, pointed out that it was some time before the working party was due to report and even then it would only report with draft rule changes. But she added: "We welcome anything that can be seen as a progressive move."


text: Shopping & Services Catching Fireflies Unique Gift Boutique in Southeast Michigan « Previous Next » AnnMarie Ottoy • April 22, 2013 Add Comment Total: 0 0 0 0 0 0 0 Looking for something out-of-the-ordinary, but not sure what exactly that may be? Buzz into Catching Fireflies – a neat indie boutique that now has a downtown Ann Arbor location to complement its two Oakland County stores. Owners April and Steve McCrumb invite you to peruse their plentiful collection of whimsical, often handmade, wonders, for kids (from babies to teens), teachers, grandparents – and you, too! A welcoming scene Describing herself as "the creative half" and her husband, Steve, as "the business half," April and her staff comb gift shows to unearth the creations suited to her particular brand of whimsy. Catching Fireflies Ann Arbor: 419 Detroit St., 734-531-6293; 10 a.m.-6 p.m. Monday-Friday, 9 a.m.-6 p.m. Saturday, 11 a.m.-5 p.m. Sunday Berkley: 3117 W. 12 Mile Road, 248-336-2030; 10 a.m.-6 p.m. Monday-Saturday (till 8 p.m. Thursday), noon-5 p.m. Sunday Rochester: 203 E. University Drive, 248-650-3318; 10 a.m.-6 p.m. Monday-Saturday (till 8 p.m. Thursday), noon-5 p.m. Sunday Bathtub teas, bacon-flavored toothpicks, cupcake go-go purses and floral jewelry pouches are just a few examples. "I feel really grateful to be doing this for a living," April says. Her customers inspire her, as well. She enjoys seeing a face light up when someone finds the perfect gift. There are many to chose from, too, in categories such as trinkets and treasures, wine lovers, Michigan goods, kitchen, office, travel, wedding and jewelry – to name a few. Spreading their wings No stranger to the community, April began selling her handmade photo frames at The Sunday Artisan Market in Ann Arbor while she attended Eastern Michigan University. "I've always loved the area," she says, and had been keeping her eye on the Ann Arbor building – which is nestled near Kerrytown and Zingerman's Delicatessen – for some time, feeling it would be a good fit for the store. The third in a growing chain of stores, the latest Catching Fireflies opened in February of 2013, with an official grand opening in March. Its original Berkley location opened in 2000, and it has a Rochester nook, too. Products are also available online. See something you like? You can have it shipped to the store and pick it up from there.


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EA_Photogr... Photographer
Timeshiana WindleyModel Female Greensboro, North Carolina, US
Mayhem # 2948704
About MeGreetings,
I am currently a college student at NC A&T State University, but I am from Virginia. Therefore, I do not mind traveling for shoots and from May to August I am primarily located in Virginia. I am very openminded to all different walks of fashion and easy to work with. I have some experience as far as a couple local shoots and hair shows, however I am willing to learn new things. I am excited about joining MM and ready to build my portfolio up by shooting with some new photographers and networking. One day I hope to become a high fashion editorial model by taking on shoots one by one. The dedication, passion, confidence, and look is there, I just need it to be captured! Contact me for any future bookings or information.
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With the advent of the era of big data, how to manage the data becomes a serious problem in the business world.Many companies have large amountsof data but struggle to analyze them to get the most benefit. A recent study by Gartner Research indicates that less than five percent of e-commerce companies use big dataor predictive analytic software. Typically, only the biggest companies such asAmazon.com, China's Alibaba and Japan's Rakuten are serious about putting in the money to make this happen.Currently, most of the data is only used to identify trends and increase sales by driving morecustomers to their online stores. As such there are great opportunities for academics to analyze the data in different ways to extract information useful to companies to improve their competitive position.
With the rapid development of the Chinese economy in recent years, a large number of enterprises with extraordinary vitality have emerged such as Huawei, Alibaba, and Baidu. These enterprises have grown because they draw on the outstanding experience of foreign enterprises but at the same time they engrave deeply the characteristics of Chinese culture into the business model. To understand the success of these emerging Chinese enterprises better, we need to develop a new research perspective.
The objective of the conference is not only to promote the development of theory and technology in the field of decision sciences but also create an excellent academic platform for international scholars to exchange research ideas.
Guanghua School of Management, Peking University
Haidian District, Beijing
Beijing is the capital of China and is the third largest city in the world with 21 million people. The city is the political, cultural, and education center of China. Beijing has the second busiest airport in the world by passenger traffic. The city has seven UNESCO World Heritage sites – Forbidden City, Temple of Heaven, Summer Palace, Ming Tombs, Zhoukoudian, Great Wall, and Grand Canals.


text: Divxturka.net is using Redbunker! Because Redbunker is better than Hotfile, Netload and Rapidshare ! Click here and buy REDBUNKER premium account! Plot: Carmen Vega is notorious for meddling in her grown daughters lives, and her new project is mapping out a master plan to find a husband for her eldest daughter Ally. The career-minded Ally is more interested in her job as an architect until she falls for Ben McInnes, the new project manager at work. Carmen takes a liking to Ben, but the real question is will Ally and her new suitor survive all of Carmen's plotting to make sure they ride off into the sunset together! Genre: Romance IMDB Rating: 4.9/10 from 46 users Directed by: Patricia Cardoso Starring: Sonia Braga, Mercedes Renard, Ana Ayora Release Name: Meddling.Mom.2013.DVDRip.XviD-EVO Size: 700 MB Video: AVi | 624×352 | 1019 kbps Audio: English | MP3 | 128 kbps Runtime: 1h 24mn Information: Code: http://www.imdb.com/title/tt2256761/ Download Meddling Mom 2013 Code: Single Link No Extraction http://www.redbunker.net/ksyvvflftcxa/Meddling.Mom.2013.DVDRip.XviD-EVO.avi.html IF YOU LIKE MY UPLOADS PLEASE GET A REDBUNKER PREMIUM ACCOUNT, Thank You All for your Support Screens: Enjoy!


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Ivivi Lodge
No prices found through our partners. Please contact the business directly or check some of our recommended alternatives.
TripAdvisor Reviews Ivivi Lodge Lake Lure
Travel Blogs from Lake Lure
Vaccines, Shots, First Aid Kits & Mosquitos
We went to the local county Health Department today to have another series of shots ---- Hep B and Typhoid. Hep B is given in 3 shots. I will need another after 4 weeks and the last one within 4 - 6 months. Typhoid vaccine lasts for 2 years. previously had Hep A vaccination and a booster shot and updated my tetanus. We are not getting Yellow Fever or ...
Clothes, Clothes, Clothes...What to bring
Now that we know what size backpack we will have -- 46 liters whatever that means, I have to fill it. Need light weight, easy care clothes -- wrinkle free. Just bought a pair of real light weight Nike's. I really like those short skirts with the shorts underneath. They are light weight, wrinkle free and ...
Backpacks and Argentina
So we need a new backpack. Have to find the largest carryon size so that we can take sufficient clothes. Tried Tortuga, but the reviews say the seams and zipper break....Not good. So back to REI for an Osprey backpack. Now to figure out what to take.....
We are booked for Argentina. We leave Chile on Dec 2 flying into Buenos Aires (BA). We booked an Air BnB from 12/2 to 1/8. We thought it would be interesting ...
Mother's Day
Got up and showered. Had just toast so pills could be taken. Went to Deer Park Restaurant in Biltmore Estate for brunch. Very, very nice and delicious......126.89 visa. Drove around the grounds and then back to the Leprechaun to take naps. Back to Leprechaun. Had a lazy afternoon. Still full from brunch buffet so had toast with baby swiss cheese and chai tea for supper......perfect. Played one game of cribbage and went to bed at 9:00. Cost of 2 sleeps at Asheville ...
A Busy Day
... I was hoping for history on the building of the road and a good look at the road itself. Did you know the Blue Ridge Mountains are older than the Alps or the Himalayas? … or that the New River (think 'The New River Train' - a song that Doc Watson and probably others recorded) is perhaps the oldest river in the world? This last fact I learned yesterday from the musicians at the Blue Ridge Music ...
Other places to stay in Lake Lure
Fox Run Resort180 Herman Wilson Rd, Lake Lure | Hotelfrom $156
The 1927 Lake Lure Inn and Spa2771 Memorial Hwy, Lake Lure | 3 star hotelfrom $99
- Credit card double charged
- Credit card trouble
- Needs some work and a good cleaning
Geneva Riverside Lodging3147 Memorial Hwy, Lake Lure | Hotelfrom $79
Fairways of the Mountains180 Herman Wilson Rd, Lake Lure | Hotelfrom $172


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You missed it This great deal for The Range at Lake Norman is over. But don't despair, we've got more great deals every day.
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The Range at Lake Norman
Head to the Range: All-Day Handgun Shooting Package with Lane Rental, Handgun Rental & More
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PAID VALUE EXPIRES ON May 25, 2022
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Hwy 10 North, Mono, ON L9W 5N9
+ taxes & licensing
Popular Ford Ranger. Auto, ext cab. Certified and etested. Price plus taxes and licensing.
This vehicle is Safety Certified and E-Tested.
Please contact the seller to confirm pricing, features, odometer, and availability of this vehicle.
See the Carpages.ca Terms & Conditions for more details.


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Daniel Audette
View Single Post
Call me Suds
Registered User
Join Date: Dec 2012
Posts: 164
vCash: 500
Originally Posted by
hab 4ever
I think it is too early to write him off as a NHL prospect. I believe his skillset is much, much better than many short players in the QMJHL. What if he is a top-20 scorer in the league next season at 17 years old and at 5'9" 180? That could still be second round pick material.
Did I miss something? I said his NHL potential was limited unless he grows in the post you quoted and you're post says he could still have NHL potential if he grows.
You said what I said in a different order.
As for the top 20 scorer thing, that doesn't always translate to NHL potential or recognition. Another good example is Nicolas Petan. He's considered a 3rd or 4th round pick right now and he's a '95 born in the WHL with 60 points in 35GP (24G 36A) with a +38 because he's only 5'9" 170lbs At that size, it's tough to make it and there are plenty of small guys who haven't despite being very productive in the CHL.
All you can do is hope he grows or develops a skill set and strength that makes his size irrelevant and Audette isn't even close to that caliber right now.
Call me Suds
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Bentley Organic Organic Baby Oil with Sunflower, Jojoba & Chamomile - 250ml
Bentley Organic »Product code: 297093
Purchase Options
Earn 33 reward points on this item Gift wrap available
Product Description
This organic baby oil is part of the fabulous certified organic baby care range from Bentley Organic. It's formulated with sunflower and jojoba to moisturise and protect delicate skin and conditioning chamomile to leave your baby feeling soft and smooth. This effective moisturiser is easy to apply, smells great, absorbs easily into the skin and keeps baby's skin soft until next bath time. Perfect for irritated skin conditions and itchy skin in both mother and baby alike.
There are currently no reviews to display for Bentley Organic Organic Baby Oil with Sunflower, Jojoba & Chamomile - 250ml
There are currently no questions to display about Bentley Organic Organic Baby Oil with Sunflower, Jojoba & Chamomile - 250ml.
About Bentley Organic
Bentley Organic are passionate about natural skincare products and they pride themselves on using only the finest ingredients including certified organic oils and spices that are environmentally sustainable and conform to certified standards.
Bentley Organic products embody the essence of nature, using only the highest grade oils and extracts to stimulate the senses and pamper your skin. They maintain the highest levels of care to ensure all of their natural, plant-based products remain free from harmful chemical additives and are not tested on animals.
Choosing organic skin care and organic beauty products is a and can also provide some relief for many common conditions such as eczema, dermatitis and psoriasis. Bentley believe that you shouldn't have to pay more for organic products and are very proud of their affordable range of skincare.
Bentley Organic is a Soil Association certified producer of personal care products. This certification not only confirms their commitment to providing the highest quality skincare products but also acts as your guarantee that all of their products are made using Organic Agricultural Oils & Spices.
Product Ethics & Tags
The Bentley Organic Organic Baby Oil with Sunflower, Jojoba & Chamomile - 250ml product ethics are indicated below. To learn more about these product ethics and to see related products, follow the links below.
Please see our Despatch & Delivery page for more information on delivery areas, services and charges.


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(678) 693-3483(678) 693-3483
4371 Winters Chapel Rd, Atlanta, GA 30360
I used these guys for my move and I tell you they are pros! They know all the angle tricks, protect your goods,and make your move quick and time consuming, these guys don't lay around! Will recommend.
Posted: 8/27/2013 • Link to this review


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Compare Business attorneys near you. Read reviews, see Avvo Ratings, and contact them directly.
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5.0 stars 11 reviews
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Attorney Donnie O'Pry has been repeatedly rated as one of the Top Lawyers in Louisiana by multiple publications including Acadiana Profile Magazine, Superlawyers Magazine, the Times Picayune and American Lawyer Media. Donnie holds the highest rat...
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Con has a wide range of experience with business transactions, commercial litigation, federal and state tax law, and wealth transfer planning. He serves clients in various business sectors including energy, healthcare, real estate, and internet te...
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Learn everything you need to know to enter the world of non-profits and writing grant proposals.
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1 recipes by pacomati
- Middle Eastern FattehI learned this from a Lebanese friend. In the Middle East, it's like the equivalent of a nacho dish. An easy and light vegetarian dish.Garbanzo beans (boiled until tender, or canned), Plain yogurt, Garlic (grated), Salt and pepper, Lebanese bread (or tortillas), Pinenuts, Butter


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If you think the price is too high, please click to apply for lower price
|5 Piece
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|5 x Screen guard films
1 x Cleaning cloth
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ótimo produto e com boa aparência, vale a pena comprar, se adapta bem ao celular e proporciona conforto
Buen protector mate, merece la pena
Pros: Good priced screen protectors, protect the device well. The matt finish is nice and does not seem to impact the screen sensitivity or view ability. Cons: Some of the punch outs are not pushed through. Have to cut out... Read more
Cheap! Good quality! Quick shipment! 6 months with the same screen protector and keep going! thanks
- Good anti-reflection capabilities - Comes in a package of 5 screen protectors - Decent quality - Fits the Nexus 5 well - Very easy to apply to the screen - Somewhat grainy when applied (but this is expected) - Bubbles be... Read more
This screen guard/ film protectors are awesome. There are 5 screen guards for a very little price. The quality of the arm matte is very a... Read more
ótimo produto e com boa aparência, vale a pena comprar, se adapta bem ao celular e proporciona conforto
Buen protector mate, merece la pena
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Style# 2001338
$64.99 $34.98
Additional Information
Print this tab Put in some extra steps with the Skechers Go Flex Walk Shoes. Built with soft knit fabric uppers for comfort and flexibility, these shoes feature an easy slip-on design and memory foam heel padding. With additional Goga Plus+® footbed technology, you'll feel like you're walking on clouds.
- Soft knit fabric upper
- GO FLEX™ articulated segmented sole design provides anatomically correct ultra-flexibility
- Resalyte® sole - proprietary lightweight injection-molded compound with memory retention helps absorb impact
- Goga Mat® technology
- Features high-rebound cushioning with responsive feedback
- Memory Form™ Fit design with memory foam padding around heel for a custom-feel fit
- Flexible rubber sole provides durability and stability
- Synthetic heel panel with Quick-Fit Portal
Print this tab
Tech Specs
- Width: Regular
- Gender: Women's
Miscellaneous Attributes
- Country of Origin: Imported


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This new born baby was found dead inside a canal after it was dumped by the mother shortly after she gave birth.
Newborn baby found dead inside dirty canal with umbilical cord still attached
By Sugar Daily 3 2016-11-29T20:10:00+01:00 Heartbreaking Photos , Say What?!?
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Once again it's time to pull on your Pro Cycling Manager jersey and to sit down in front of the world's top cycling game. Now in its 7th edition, Pro Cycling Manager continues to evolve while still offering those elements that have been key to the game's success over the years :
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Name of threat: EduCrypt Ransomware
Category: Ransomware
Threat level: Very high
Affected countries: USA, Germany, France, Italy, Spain, Russia, China and others
Affected OS: XP, VISTA, 7, 8 and almost 10
Encrypted files: .txt, .doc, .png, .jpeg, .ppt, .xls, images, videos and others
EduCrypt Ransomware is categorised as very nasty and dangerous ransomware program which causes lots of big issues in your computer, once it get installed. It is capable to silently enter into targeted victims PC for the encrypting of files and data without taking your permission and approval. It encrypted all useful files such as text files, documents, PDF registry and etc. It told you that your files get corrupted due to some illegal works from your IP address such as pornography, dating site, click on unsponsored links, fakes updates of application and lots of more. It suggest you to pay some ransom money through the BITCOIN for the given certain time and get the decryption key. Otherwise, if you delay for the payment of money then it can transfer these all information of head branch of cyber-crime office that will take heavy charge for doing this penalty.
According to member of cyber experts, EduCrypt Ransomware is generally a fake and bogus infectious program which aim is to only fool innocent user for the cheated of their privacy and data. It is mainly intrude in your computer with the bundled of file sharing, spam email attachments, peer to peer to file share, downloads game, movies torrent files and lots of more. Apart from these, it can also invade its malicious extension toolbars in your browser like add-ons and plug-ins. It can easily track your browsing experience including searching history, cookies, saved bookmarks page, Online transaction details, bank account details and lots of more. It may transfer that information to the remote cyber criminals that may use it for their illicit purpose. Therefore, you must be suggested to eliminate EduCrypt Ransomware instantly from your computer to keep safe it for the long time in future.