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The hyperfine structure of the ground state $4f^76s^{28}S_{\frac{7}{2}}$ of $^{151}$Eu and $^{153}$Eu has been measured by the method of magnetic resonance in an atomic beam. The sign of the hyperfine structure and the value of g$_J$ have also been measured. The results are: $g_J = 1.9935 \pm 0.0003;$ $A(151) = -20.0523 \pm 0.0002 Mc/s, A(153) = -8.8532 \pm 0.0002 Mc/s,$ $B(151) = -0.7012 \pm 0.0035 Mc/s, B(153) = -1.7852 \pm 0.0035 Mc/s;$ $\frac{A(151)}{A(153)} = 2.26498 \pm 0.00008,$ $\frac{B(151)}{B(153)} = 0.393 \pm 0.003.$ These results disagree with the values expected for the pure $^8$S$_\frac{7}{2}$ state formed from a half-filled shell of f-electrons. Calculations on the basis of admixture of higher states of the configuration by spin-orbit coupling account for the discrepancy in g$_J$, but do not explain the values of the hyperfine structure constants A and B. Further theoretical work is in progress.
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1) Unavailability of students in VHSE and vacant seats in HSE – VHSE is always considered by the public as a last resort for education in plus two levels. Schools which are conducting VHSE courses are always finding difficult in getting students for every year. (Most of the schools which I know hardly get 50% of students each year that too at the last moment before the admission closes). If a survey is conducted among the Students studying in VHSE course, it will be clear that most of them may not be even aware of the importance of the course they are studying. On enquiry most of them replied that they don't get seat in HSS and hence opted VHSE
2) It is interesting to say that the total number of seats which remains vacant in HSE in each year is more than the total number of students studying in the VHSE in the whole state.
3) Deviated objective of VHSE – The objective of Vocational Higher Secondary education is to make a student capable of handling a technical skill with out further external support. As at present it has much diverted from the actual objective. Present system adopted by the State Government in VHSE is just equivalent to higher secondary Courses with addition of vocational courses in it. In many schools it has been seen that the importance of the Vocational subject is totally neglected giving more importance to non vocational course there by nullifying the importance of VHSE.
4) A student who completes the VHSE course must be able to handle the technical skill he acquired with out much external technical support. But now most of the students who join VHSE are just joining as a last resort and such students never acquire any technical skills after completing the course and they go out empty handed after (Better say wasting) two years.
5) Further the responsibility of the VHSE department finishes once the students completes the course and a certificate issued on successful completion of the course. There is no follow up on the students in the way of performing the skill they acquired. In most of the cases it is seen that after completing the VHSE course, the student will go for general degree course, which never have any connection with the skill they acquired through completing VHSE course. This justifies the saying that they don't get seat in HSS and hence opted VHSE
6) Outdated courses - Most of the Vocational Courses offered to students at present are outdated. To cite just an example, the Type writing skill associated with the course Office Secretary-ship is a mere waste in the present scene. When computer has replaced Typewriter and almost any one can handle a computer now a days, what relevance does it make with the course?. Another one course in Agriculture (Plant Protection) does not offer any job even to the brightest student who studies this. Then what is the use of such courses?
7) Work load comparison – If we compare the work load of VHSE staff it has been seen that it is much lower than the work load of Higher secondary staff (I am saying about the actual work load and not the written work load) The Vocational staff may be afraid of losing the "Benefits" of low work load enjoyed by them, once they are reallocated to other departments or in the same department itself with different nature of work there by resisting the new system.
Irresponsible teachers - Now a days It has been seen that most of the time the teaching staff instead of performing their duty are seen engaged in surfing free internet social sites or discussing family matters. Some of them are found engaged in activities for improving side income.
9) Neglecting the importance of Marketing in curriculum – Since this is a job oriented program, it is true, production is considered great importance and that is what is being taught. But the marketing part is totally neglected in the curriculum. In the present scene when the success of any product is based on the marketing a product, it is worth mentioning here the irresponsibility of the VHSE department by neglecting the importance of marketing
10) Inequality among HSE and VHSE students – By offering technical skills only to a group of students, our government is denying opportunity to learn these skills to other clause of people. Is this not against the Right for Education Act ?
Hence my suggestion is that both departments (VHSE & HSE) should be merged together and the excess staff must be redeployed accordingly, Major suggestions includes,
1) Many Vocational staff working in the Vocational Higher Secondary Education Department is interested to get reallocated in other departments. Hence option may be called for from the interested employees of VHSE to get reallocated/ Redeployed
2) Most of the Non Vocational teachers don't have any resistance against the new proposed system of merging VHSE department with HSE department because they themselves likes to be known as a part of the Higher Secondary Education Department rather than the VHSE Department (This is evident from the employees turn over of the department to HSE Dept.) Only some vocational Staff is resisting the change in to new system of merging. To limit their resistance it is better to start implementing the new system in stages. As the success of any system depends on the mentality of Human Resource in it, only those who are interested in reallocation should be included in the first phase
3) The VHSE may be stopped in a phased manner in stages to incorporate the new scheme (Vocational Education and Training Center– VET). For this the new system has to be introduced in an experimental basis in this academic year (2012-13) itself, study the drawbacks, make necessary changes and implement in full fledge from the next academic year onwards
4) Counseling the Vocational Staff for fear of unemployment – Most of the Vocational Staff misbelieve that they may be thrown out unemployed when the new scheme is introduced.
5) The case of handling the VHSE staff in aided Management Schools should be thought of separately
There may arise difference in opinion and resistance from the part of the Staff of VHSE department as there was when Pre-Degree was delinked from the Collegiate Education Department. As a responsible minister in this Government, I believe it is your duty to safe guard the future of the students and our state as a whole rather than protecting the selfish motives of some staff in the VHSE department

subset: commoncrawl


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Peppermill Resort 2707 S Virginia St
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The Barry M. Goldwater Scholarship and Excellence in Education Program was established by Congress in 1986 to honor Senator Barry M. Goldwater, who served his country for 56 years as a soldier and statesman, including 30 years of service in the U.S. Senate. The purpose of the foundation is to provide a continuing source of highly qualified scientists, mathematicians, and engineers by awarding scholarships to college students who intend to pursue careers in these fields.
The purpose of the Barry M. Goldwater Scholarship and Excellence in Education Foundation is to alleviate a critical current and future shortage of highly qualified scientists, mathematicians, and engineers. The foundation will award up to 300 undergraduate scholarships to outstanding students, to be known as Barry M. Goldwater Scholars.
Students who plan to study medicine are eligible for a Goldwater Scholarship if they plan a research career rather than a career as a medical doctor in a private practice. Applicants should have a strong commitment to research and be able to articulate specific research questions to which they intend to devote their professional life.
Each scholarship covers eligible expenses for undergraduate tuition, fees, books, and room and board, up to a maximum of $7,500 annually. Scholarship monies not used during one academic year are not transferable to the succeeding academic year.
UM students seeking nomination for the Barry M. Goldwater scholarship must submit the preliminary application to gain access to the full application. The full application, including letters of recommendation, is due by the campus deadline. The University of Miami may nominate no more than four students to compete for the national scholarship. A selection will be held to identify the nominees whose applications are most likely to fare well in the national competition. Students with a strong academic and research background are encouraged to apply. The Goldwater scholarship is intended for students planning careers in research (Ph.D. tracks), rather than careers as practicing medical physicians. Click on the link below for instructions on applying for Goldwater nomination as a UM student.
Barry M. Goldwater UM Student Application Instructions
**The PAF office will only provide feedback to applicants who request it and who submit their materials for review a minimum of three weeks in advance of the campus deadline. NO EXCEPTIONS.**
September 2016
Online application and materials available from the foundation's website.
November 7, 2016
All application materials must be submitted to the Office of Prestigious Awards and Fellowships for review and possible endorsement.
January 18, 2017
Deadline for online submission of UM nominee applications.
Late March 2017
Nominees and faculty representatives are notified of applicant status.


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934 acres in Kimble County, Texas
Located only 3 miles from Junction and the Kimble Co. airport, this ranch has breathtaking views and one mile of seasonal creek. Improvements include a 3/2 house, 2 steel barns, a cantina, several blinds and feeders, and a storm cellar/wine cellar. There are 2 wells on the property. A large population of deer and turkey provide good hunting.
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Greer or Kelly Kothmann
R.D. Kothmann Real Estate
1701 Main St
Junction, TX 76849
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NOTE: The information contained herein is provided as general information only and has been obtained from sources deemed reliable. It is provided without any guaranty, warranty or representation, expressed or implied, made by Lands of America, or any related entity, as to the accuracy or completeness of the information. The information is presented subject to errors, omissions, change of price or conditions, prior sale or withdrawal without notice. Prospective purchasers should make their own investigations, projections and conclusions concerning the information.


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Cyprus Escorts > Limassol Escorts > Anna Escort
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Data from: Bacterial motility confers fitness advantage in the presence of phages
Date Published: June 13, 2013
DOI: http://dx.doi.org/10.5061/dryad.t7t54
Content in the Dryad Digital Repository is offered "as is." By downloading files, you agree to the Dryad Terms of Service. To the extent possible under law, the authors have waived all copyright and related or neighboring rights to this data.
|Figure 2a and b Area Dispersed
|39 times
|Data shows the colony area of Pseudomonas aeruginosa after 48 hours growth and dispersal under different phage treatments (no phage, local phage and global phage), and different agar types (hard and soft). Data was collected in vitro and excel was used to create data file.
|Figure 2a and b Area Dispersed.xlsx (10.69 Kb)
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|Figure 3 Relative fitness
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|Data shows the relative fitness of dispersers to non-dispersers (i.e. CFU disperser/ CFU non-disperser) in Pseudomonas aeruginosa after 48 hours growth and dispersal. Motility variants were tested across three treatment groups: No Phage, Local Phage and Global Phage. Excel was used to create the data file.
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I stayed here based on the fact that I was attending a conference down the street at the HI Express and this was a better deal. When I checked in the staff was pleasant and helpful. I got a room on the 3rd floor and when I walked in, it smelled like someone had just smoked a pack of cigarettes in the room. I went back down to the front desk and inquired about it and was immediately offered a room swap, which I took. The next room was great and I was pleasantly shocked to find NBC Sports was offered on the TV so I was able to watch the Stanley Cup Finals, bonus! Pretty standard Comfort Inn room. It was clean and quiet. The bed was too hard for me and my bad back, but that's pretty standard. The internet worked great and I was able to get my work done while staying there.
Overall the hotel was fine. It wasn't anything to ooh and awe over and it did the job. It's conveniently located close to the freeway and you are not too far from town. My fellow conference attendees all stayed at the HI Express and were not impressed at all. Plus they paid double what I paid. I would stay here again.
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Celeste and Jesse Forever
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Plot: Two people who met in high school and married young are growing apart. Now 30, they decide to divorce and attempt to stay best friends while pursuing other relationships. This is not as simple as they had imagined, however. Celeste - imperious, successful and driven - is convinced that divorcing Jesse is the right thing to do, a decision Jesse accepts although he is still in love with her. As the reality of their separation sets in, Celeste realizes she has been cavalier about their relationship, but the timing of her revelation is less than fortuitous. While navigating the turbulent changes in their lives and in their hearts, these two learn that in order to truly love someone, you may have to let them go.
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Harrison Brook financial advisers is your perfect partner. Harrison Brook is the leading international financial consultancy of expatriate financial service advice and products to expatriates.
We provide the expat community the best value UK pension transfer advice, retirement planning solutions, investments (income generation/capital growth portfolios) and international insurances from the world's leading investment firms.


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Houses could be built on open farmland in Fleet, opposite the sports centre, if a developer's appeal is successful.
The developer has launched an appeal because Hart has failed to make a decision on its plans for building homes on land west of Hitches Lane.
Hart says it has been unable to consider the plans while it waits for a crucial report on land development in the area.
Last March Hart published a document outlining possible land available in the district for both recreational and residential use. Planning applications relating to these modified plans and including Hitches Lane have not yet been considered because members were awaiting the "Urban Capacity Study" report.
In the report, land suitable for development would be identified and preference given to available, suitable, urban or previously developed land. After that, greenfield sites needed to fulfil district housing requirements up to 2006 would be looked at followed by a revised Hart District Local Plan identifying land earmarked for development.
Coun. Colin Hazell said it was disappointing that Berkeley Strategic Land was placing extra work and cost on Hart when the delay in processing its planning application was due to Government requirements.


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Tyrone Branch
This Branch now normally meets in Omagh at 10.00am on the second Saturday of the month October – May.
Meetings are held in the Seminar Room, Omagh Library, Dublin Road, Omagh, BT78 1HL.
Annual Subscription
for the year 1st September 2016 to 31st August 2017
- £15 for Single Membership
- £20 for Joint Membership (2 people at the same address)
Officers 2017-2018
- Chairperson: Vincent Brogan
- Secretary: Deirdre Muckle – E-mail: firstname.lastname@example.com
- Treasurer:
Social Media
Omagh Branch has a Facebook Group.
Saturday 3 September 2016 (1st Saturday)
Registration and "Social Media and Genealogy"
Sheena McClure
Wednesday 5 October 2016
Visit to the LDS Family History Centre, Racecourse Road, L/Derry – 7pm
Keith Wright
Saturday 12 November 2016
"A Tale of Two Cemeteries – a poignant story of two forgotten girls"
Harman Scott
Saturday 10 December 2016
"Ower the Sheugh" – considering the Irish and Scottish migrations across the Narrow Sea in historical prespective.
Dr Paddy Fitzgerald
December 2016
Meal out with family and friends. Contact Dee Muckle to confirm details.
Saturday 14 January 2017
"The Huguenot Jack Family of Ardstraw 1360-2015"
Frank Collins
Saturday 11 February 2017
"The Evolution of Surnames"
Vincent Brogan
Saturday 11 March 2017
"Neighbours, Landlords and Tenants on the Mountjoy and Abercorn Estates"
Dr Brian Lambkin
Saturday 8 April 2017
Annual General Meeting followed by short talk on updates in DNA for family history
George Gordon
May 2017
A group outing, date, time and place to be discussed and arranged nearer the time.


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South West & Wales Anti-Fracking Gathering in Weston Super-Mare
March 19, 2016
On 19th March 2016 Frack Free Somerset invite you to gather for a day of networking, sharing & learning. We are planning an interactive event with the opportunity for everyone to participate in focussed open discussion spaces.
Speakers include: Theo Simon of Seize The Day, Tina-Louise from Lancashire, alongside people from Reclaim The Power & Frack Off. There will also be a breakout room for chilling, films, rugby score catch up's, tea, etc.
Topics include: Underground Coal Gasification, Groundswell, What can we Do about Fracking, the regional picture of extreme energy in the SW and Wales & more… Topic suggestions are welcome 🙂
Simple lunch & beverages will be provided & there Will be Cake
To book your space please contact email@example.com


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January 10, 2011
Jae Millz celebrates his video release party at Club Play with Mack Maine and the Young Money crew, along with Trey Songz, @Aubreyanne (who I finally got to meet), Pretty Jamie, Spectacular, Jasmine Calle, Cassie Melinda, DJ Stevie J, Top Feelin, and more. Brought to you by Headliner Market Group. Press play to see what went down.
And next Friday, check out Bow Wow at Play as well for part two of the Young Money takeover.


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CBT and Ballbusting
CBT And Ballbusting 76
Elise Graves has her gimp leaned over rearwards and fastens strenuous weights on a pulley to her sub's testicles. This is his highly very first COCK AND BALL TORTURE practice, so he has never perceived this kind of ache before. He came back for more as shortly as he could.483 Views
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When Spiceworks pulls from AD it does not popluate the users 'supervisor' field so when I try to enabel the AD sync so Spiceworks can send changes back to AD it tries to clear the field in AD.
This topic was created during version 5.1.
The latest version is 7.5.00101.
5 Replies
Jul 31, 2011 at 5:38 UTC
GreyRose LLC is an IT service provider.
The AD info you refer to is populated from the Organization tab in the AD user object. Is that correctly input already in AD?
Jul 31, 2011 at 5:56 UTC
Yes the field is already populated in AD. The spiceworks sync back wants to clear it because the version of the user record that was created from the initial AD pull is blank in spiceworks.
Jul 31, 2011 at 6:54 UTC
GreyRose LLC is an IT service provider.
That's unusual. How many people in AD?
Jul 31, 2011 at 7:13 UTC
About 60 users
Jul 31, 2011 at 8:22 UTC
GreyRose LLC is an IT service provider.
That's really unusual. If it was a lot, I might consider that the data sync was incomplete. for 60 users, though... You may want to email email@example.com as that's a pretty unique problem.
This discussion has been inactive for over a year.
You may get a better answer to your question by starting a new discussion.


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: 11 November, 2009
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New Backup and Archiving support available from Symantec for Microsoft Exchange Server 2010 featuring granular recovery and rapid search capability
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Symantec has announced updates to its backup and archiving software solutions to support Microsoft Exchange Server 2010 and other Microsoft applications. Symantec's information management solutions help extend Exchange 2010 by further protecting customers' information, reducing storage and server costs and quickly recovering information for business, legal and regulatory compliance purposes.
Symantec's Enterprise Vault archiving solution adds direct drag-and-drop access to the archive from Outlook, so users no longer have to rely on "shortcut" links from their mailbox. This is now available for previous versions of Exchange and is scheduled to be available for Exchange 2010 in early 2010. Available now, Symantec Backup Exec System Recovery 2010 provides fast, reliable system backup and restore for Microsoft Exchange 2010. Over the next several months, Symantec is scheduled to deliver updated versions of NetBackup, Backup Exec and Enterprise Vault to reduce storage by deduplicating information stored in Microsoft Exchange 2010, as well as other applications such as Microsoft SharePoint and Windows Server 2008 R2.
"Organizations of all sizes are wrestling with massive information growth across multiple Microsoft platforms and applications, but especially Exchange," said Mathew Lodge, senior director of product marketing, Symantec Information Management Group. "Symantec helps by providing backup and archiving platforms to centrally protect, manage and discover critical information within Exchange 2010 and other Microsoft platforms."
Symantec Enterprise Vault unifies deduplication, tiered storage, policy enforcement and e-discovery of information across multiple applications, such as Exchange, SharePoint, file servers and instant messaging. Available now, Enterprise Vault 8.0 with virtual vault eases the end user experience for organizations archiving Microsoft Exchange by combining access to live, legacy and archived emails in Microsoft Outlook without any visible change to the appearance of the message or stub files. This new capability allows end users to easily drag, search, move and retrieve archived email within Outlook just as they would with a .pst but without additional complications related to e-discovery or corruption. Additionally, Enterprise Vault is scheduled to provide fully-integrated archiving support for Exchange 2010 early next year.
"Symantec has unique expertise in backup and archiving solutions, and we are happy to have them as a Microsoft partner for Microsoft Exchange Server 2010," said Betsy Frost, general manager, UC Marketing at Microsoft Corp. "Partners are critical to Microsoft's success and, by building on Exchange 2010, Symantec helps us deliver more value to our customers and meet the needs of a wider variety of organizations. We are delighted to have Symantec included in our community of partners who are energized and ready to help customers take advantage of the innovations in Exchange 2010."
Symantec Backup Exec System Recovery 2010 provides small businesses with full system backup and recovery of the latest Microsoft platforms, including Microsoft Exchange 2010, Windows 7 and Microsoft Server 2008 R2. This allows organizations moving to these new Microsoft environments to ensure that their data and systems can be quickly recovered should they encounter an outage or disaster.
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Summer Blockbuster Week
Summer BLOCKBUSTER WEEK
MONDAY, August 8
Lamb Dance | Children's Show
A children's puppet TV show directed by Rapp Battlez's Freddie Rivas.
Sex AND the City | Rom-Com
Friendships don't magically last for forty years, you have to invest in them.
ft. Bridget Cann, Alan Bee, Shohana Sharmin, Peter Stelmach, Marc Johnston, Brody Trottier & Cales Le Grand.
Epic Bedtime | Thriller
After this bedtime story, no one is sleeping.
Directed by Leigh Cameron.
Don't Do That: A Teen Message Play | After School Special
We're all choked up and ready to put the "bawl" in play.
ft. Justin Collette, Connor Low, Alex Kolanko and a very special guest coach!
PANTSUIT | Political drama
Suits. But with pants.
ft. Ivan Yuen, Devon Henderson, Brittany Drysdale, Jacqueline Twomey, Cecilia Seraphina, Hamed Dar, Christina Nicolaou, Tara Federko & Terris Taylor.
Law And/Or Order: UK | Crime
20 Minutes of Law. 20 Minutes of Order.
ft. Alistair Forbes, Alice Moran, Robb Baker, Jon Blair, Jess Bryson, Christie Bruce & Kris Siddiqi.
TUESDAY, August 9
Ghostbusters 2 | Hollywood remake
Who you gonna call? Us, to remake your movies.
ft. Sheri Anne Godda, Emily Opal Smith, Amanda Pereira, Victoria Kucher & Brennan Asbridge. Directed by Jan Caruana
Ontario James & the Temple of Doom | Action/Adventure
A last crusade to the temple of the lost ark.
ft. Scott Goldman
House of Scandal | Political Thriller
Every political legal thriller always leads to murder... sexy, sexy murder.
ft. Alessandra Vite, Allana Reoch, Ted Hambly, Sean Tabares, Christian Smith, Kevin Vidal, Lisa Gilroy & Tricia Black.
Dreamboats | Fantasy
Because dreams are just improv in your brain.
ft. Caitlin English, Samantha Chaulk, Nelu Handa, Andy Perun, Naseem Hrab, Anesti Danelis, Robbie Grant, Cassie Cao, Jennifer Ferris & Justin D'Angelo.
Arc | Genre Mashup
Destinies Collide: what happens when two characters played by the same actor meet in the same movie?
ft. Michal Grajewski, Amy Matysio, Mike Fly, Jayden Pfiefer and musical accompanist Nick Di Gaetano
Wednesday, August 10
The Adventures of Mary-Kate and Ashley | Children's Mystery
They'll solve any crime by dinner time!
ft. Lisa Gilroy and Natalie Metcalfe
Your Kids' Movie | Children's Movie
They'll get into trouble!
ft. Nadine Djoury, Kevin Vidal, Christian Smith, and Liz Johnston with possible special guests!
Twice Upon a Time | Musical
The story before the fairytale.
ft. Irina Mann, Richard comeau, Jonathan shaboo, drew Delaware, ellie Goodman, Candace Meeks, Blair Mitchell
The Nightfall Realm | Sci-Fi/Fantasy
You're travelling through an improvised dimension.
ft. Nora Boydell, Kjell Cawsey, Greg Fox, Scott Goldman, Candace Meeks, Josh Obermeyer, Laura Salvas, Mark Shyzer
Easy Action | Adventure thriller
Award-winning Montreal darlings bring their signature two-hander tour-de-force back for their fourth Blockbuster Week!
ft. Marc Rowland & Brent Skagford
THURSDAY, August 11
The One | Reality TV
Fall in love with this show.
ft. Nicole Passmore, Sam Loeb, Talal Itani
Easy Action | Adventure thriller
Award-winning Montreal darlings bring their signature two-hander tour-de-force back for their fourth Blockbuster Week!
ft. Marc Rowland & Brent Skagford
Real Life Toronto | Dramedy
Like Girls, but a comedy.
Directed by Kirsten Rasmussen.
Songbuster | Hollywood Musical
ft. Tricia Black, Kristian Bruun, Matty Burns, Ashley Comeau, Alexandra Hurley, Tom King, Stephanie Malek, Josh Murray, Nug Nahrgang and Connor Thompson
FRIDAY, August 12
Four directors. Four scenes. There can only be one super scene!
Fantastica | fantasy
Journey to a world built of your wishes.
ft. Tricia Black, Nate Callens, Beth Dunn, Nelu Handa, Connor Low, Rakhee Morzaria & Callum Wratten
The Underdogs | Sports
They'll win your heart but can they win the big game?
ft. Nina Adler, Lisa Amerongen, Tricia Black, Liz Johnston, Connor Low, Gordon Neill, Alice Stratford-Kurus & Callum Wratten
Brodown Throwdown
A massive festival-mashup featuring improvisers from throughout the week... and a whole lot of danger.
ft. Cast members from all of Blockbuster Week
SATURDAY, August 13
The Bad Dog Featured Players
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Met moord meer mans (2 items)
Welcome back! Whilst you were away we added the functionality to view auctions and bid in pounds.
Met moord meer mans: Information on the Catawiki site about Met moord meer mans, 1982.
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Turquoise is a stone that has been valued by humans for thousands of years. Native Americans appreciated this stone for its natural beauty and for its mythical power. It was used in ancient Egypt at least 3000 years ago. Important buildings in middle eastern cultures had domes and walls covered with turquoise. The Chinese, Greeks, Indians, and Turks all prized this pale blue stone.
Turquoise Formation
Turquoise is formed as a secondary mineral. This means that the minerals (copper, phosphate, and aluminum) that make up the turquoise were part of another mineral or group of minerals before they came together in this form. It is found in arid climates often forming in cavities of volcanic rocks.
Color(s): blues, greens, blue green
Notable deposits of turquoise are found in Iran, Sinai, and the southwestern part of the United States.


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Radhakrishnan.E.K1, K.C. Sivakumar2, E.V. Soniya1*
1Plant Molecular Biology Lab, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thiruvananthapuram- 695014, India
2Bioinformatics Facility, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, India
Received Date: May 25, 2009; Accepted Date: July 16, 2009; Published Date: July 17, 2009
Citation: Radhakrishnan EK, Sivakumar KC, Soniya EV (2009) Molecular Characterization of Novel form of Type III Polyketide Synthase from Zingiber Officinale Rosc. and its Analysis using Bioinformatics Method. J Proteomics Bioinform 2: 310-315. doi: 10.4172/jpb.1000090
Copyright: © 2009 Radhakrishnan EK, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Visit for more related articles at Journal of Proteomics & Bioinformatics
Enzymes of Type III polyketide synthase superfamily play an important role in the biosynthesis of medicinal natural products in plants. The PKSs generate the diversity of polyketide derivatives by changing their preference for starter molecules, the number of acetyl additions catalysed and the cyclisation of the polyketide intermediates. The amazing structural features of gingerol and related compounds of ginger (Zingiber officinale Rosc., Zingiberaceae) provide a genomic insight in to the presence of novel forms of PKS. The current study describes the isolation and characterisation of a novel of PKS from Z. officinale using degenerate oligonucleotide based PCR method. The inducible expression of recombinant ZoPKS in E. coli resulted in the formation of a protein with approximate molecular weight of 43kD. The comparative sequence and phylogenetic analysis of ZoPKS shows its significant variation from already identified PKSs. The novelty of the ZoPKS was further confirmed by homology modeling based comparative structural bioinformatics analysis. The novel form of PKS identified in the study has very remarkable amino acid substitutions at the key residues determining the starter substrate selectivity and condensation reactions and forms a genomic basis of PKS from Z. officinale to explore its potential in biosynthesis of gingerol and related compounds.
The amazing diversity of polyketide derivatives in plants is generated by a group of structurally related enzymes called as the type III polyketide synthases (PKSs). The most well known and widely distributed member of the PKS superfamily is the chalcone synthase(Winkel-Shirley, 2001). In the typical reaction mechanism, it forms the chalcone by the stepwise decarboxylative condensation of coumaroyl CoA with three malonyl coA followed by the claisen type cyclisation of the tetraketide product (Jez et al., 2001). Extensive gene duplication followed by the functional divergence is believed to have played an important role in generating the biochemical diversity of PKS superfamily. The expanding members of the family, as shown by the identification of 2-pyrone synthase, stilbene synthase, benzalacetone synthase, valerophenone synthase, acridon synthase etc., from various sources indicate that the biosynthetic potential of the PKS is just begin to be explored and many more members are to be identified from plants especially from medicinal plants (Austin and Noel, 2003).
Zingiber officinale Rosc. (ginger, Zingiberaceae) is well known for its use in traditional therapeutic and preventive medicine. The major pharmacologically active component present in Z. officinale is of the gingerol group (Ramirez- Ahumada Mdel et al., 2006). Molecular insight in to the gingerol biosynthesis suggested that enzymes similar to type III polyketide synthases are having important role (Denniff et al., 1980; Schröder, 1997). Considering the pharmacological potential of gingerol and the distribution of structurally similar compounds in other members of the family, identification and characterisation of the PKS from Z. officinale can provide a genomic basis for elucidation of biosynthetic pathway. In the present work, we are reporting the cloning and characterisation of a novel form of type III PKS fromZ. officinale. The comparative structural bioinformatic analysis suggests that the PKS identified in the study may be a prime candidate for the biosynthesis of phenylbutanone derivatives of Z. officinale.
Zingiber officinale var. Rio de Janeiro was used as the experimental material for the study. The plants were collected from Kerala Agricultural University, Thiruvananthapuram, Kerala and were grown and maintained in the experimental plant garden at Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala.
PCR and Cloning
Total RNA was isolated from young leaves of Z. officinale by Trizol method and reverse transcribed using the MMLV RT (Promega). Amplification of core cDNA fragment of PKS was carried out by nested RT-PCR using the inosine containing degenerate oligonucleotide primers as reported earlier (Abe et al., 2001). The sequences of the primers are as follows: PKS1 5'-RARGCIITIMARGARTGGGGICA- 3', PKS2 5'-GCIAARGAYITIGCIGARAAYAA-3', PKS3 5'-CCCMWITCIARICCITCICCIGTIGT-3' and PKS4 5'- TCIAYIGTIARICCIGGICCRAA-3'. The primers PKS1, 2, 3 and 4 represents the 112, 174, 368 and 380 amino acid position of typical CHS. The conserved amino acid residues at these areas provide opportunities for nested PCR based approach for isolating core sequence of PKSs from taxonomically diverse plants. The introduction of degenerate bases and inosine makes the primers to bind to functionally divergent members of PKS family. The core fragment (major part of the second exon) of PKS contain most of the residues contributing to the catalytic architecture. So the changes observed on conserved residues at these areas may have remarkable impact on reaction mechanism of PKS. The primary PCR reaction (50 µl) contained the cDNA template, 30 pmoles of each primers (PKS1 and PKS2), 1.25 units of Taq DNA polymerase (Promega), 1.5 mM MgCl2 and 200 µM of each dNTPs. The reaction was carried out for 35 cycles in a Bio-Rad iCycler using the following conditions; initial denaturation was done for 3 min at 94°C, the cyclic denaturation at 94°C for 30 sec, annealing at 42°C for 1 min and extension at 72°C for 1 min with a final extension at 72°C for 7 min. The secondary PCR was carried out using primer sets of PKS3 and PKS4 using 1 µl of the primary PCR product as template under the same PCR conditions as for the primary PCR. The PCR product was analysed in a 2% agarose gel and was purified using the GFX gel band purification system (Amersham). The purified fragment was ligated to pGEMT easy vector (Promega) and propagated in Escherichia strain JM109 (Promega). Plasmid isolation and purification was done using the Wizard Plus SV Minipreps DNA purification system (Promega). Automated sequencing of recombinant clones was carried out using Big Dye™ Terminator Cycle Sequencing Ready Reaction Kit Version.3.1 (Applied Biosystem) in the ABI 3730 DNA sequencer.
3' and 5' RACE
Gene specific primers designed from the core fragment of PKS from Z. officinale were used for the RACE experiments. The RACE ready cDNA was prepared using the First Choice RLM RACE kit (Ambion). The primers used for the 3' RACE were PKS31 5'- ATCGCCAAGGACCTGGCGGAGA -3' and PKS32 5'- ACAACCGCGGCGCGCGCGTCCTCG -3'. The primers used for the 5' RACE experiments were PKS51 5'- GATGTTGCTCGCAATGATCGACGGAA-3' and PKS52 5'- ATGATCGACGGAAGCTGGCTCTTGAGG- 3'. For the 3' RACE, the primary PCR was carried out with the primer set of PKS31 and 3' RACE outer primer and the secondary PCR with PKS32 and 3' RACE inner primer. The PCR condition used were; initial denaturation at 94 oC for 3 min, cyclic denaturation at 94°C for 30 sec, annealing at 65°C for 30 sec and extension at 72°C for 1 min with a final extension of 7 min at 72°C. The product formed in the secondary PCR was cloned and sequenced. The 5' RACE ready cDNA was used as template for the 5' RACE PCR using the primers PKS51 and PKS52. The primary PCR was carried out using PKS51 and 5' RACE outer primer followed by the secondary PCR using the primer sets PKS52 and 5' RACE inner primer. The conditions for both the PCR experiments were; initial denaturation at 94°C for 3 min, cyclic denaturation at 94°C for 30 sec, annealing at 65°C for 30 sec and extension at 72oC for 1 min with a final extension of 7 min at 72°C. The PCR product formed was cloned and sequenced.
Cloning and Expression of Full-length cDNA
Full-length cDNA of PKS from Z. officinale (ZoPKS) was isolated by RT-PCR using N-terminal gene specific primer NPKSF 5' CGCTCGAGCATATGCATCA CCATCACCATCACATGGGTAGCCTGCAGGCGATG 3'
containing the site for Nde I and six histidine residues and the C-terminal gene specific primer CPKSR 5' CGGGATCCCTACGGTATTGGTAGACTGCGTAG 3' containing the site for Bam HI . The PCR was carried out with an initial denaturation for 3 min at 94°C, cyclic denaturation at 94°C for 30 sec, annealing at 65°C for 30 sec and extension at 72°C for 2 min with a final extension of 7 min at 72°C. The PCR product formed was cloned, sequenced and analysed by BLAST programme. The sequence was submitted to NCBI under the accession number DQ486012. The phylogenetic analysis of ZoPKS was carried out along with other PKSs by NJ and MP methods implemented in the MEGA3 (Kumar et al., 2004).
For the prokaryotic expression, the ZoPKS was cloned at the BamHI/NdeI site of pET-32b (Novagen). The recombinant clones were confirmed for the sequence and transformed in to E. coli BL21(DE3)pLysS. The cells with the recombinant plasmid were cultured to an OD600 of 0.6 in Luria-Bertani medium containing 100 mg/ml of ampicillin at 28°C. The expression of the recombinant protein was induced by the addition of 0.5 mM isopropyl thio-β-dgalactoside by incubating at 28°C for 16 h. Western blotting of the recombinant ZoPKS was carried out using antibodies specific to His-tag (Sigma) (Towbin et al., 1979).
Multiple Sequence Alignment and Homology Modeling
Sequences of 14 plant PKSs were collected from the NCBI database and was used along with ZoPKS for the comparative structural analysis. Most of the selected PKSs were biochemically studied and were shown to have role in biosynthesis of plant specific metabolites. The sequences selected were Ruta graveolens - Acridone synthase (S60241), Aloe arborescens - Octaketide synthase (AAT48709), Aloe arborescens - Pentaketide chromone synthase (AAX35541), Arabidopsis thaliana - Chalcone synthase (CAI30411), Gerbera hybriba - 2-pyrone synthase (CAA86219), Humulus lupulus - Valerophenone synthathase (BAB12102), Hydrangea macrophylla - Coumaroyl triacetic acid synthase (BAA32733), Iris germanica – Chalcone synthase (BAE53636), Oryza sativa - Chalcone synthase (CAA61955), Rheum palmatum - Benzalacetone synthase (AAK82824), Vitis vinifera - Stilbene synthase (ABJ97068), Wachendorfia thrysiflora - Polyketide synthase (AAW50921),Medicago sativa - Chalcone synthase (P30074) and Dendrobium nobile - Chalcone synthase (ABE77392). The multiple sequence alignment of the sequences was done by using the Clustal W programme (Thompson et al., 1994).
Homology model of ZoPKS was generated based on the crystal structure of CHS from M. sativa (PDB id – Ibi5) and 2-PS from G. hybrida (PDB id –1qlv) and were taken as the representatives of the typical chalcone forming and nonchalcone (Pyrone) forming members of the PKS super family. X-ray crystallographic information of both the templates was collected from the Protein Data Bank (Berman et al., 2000). The alignment between the sequence of ZoPKS and the structure of selected templates was carried out using Clustal W as the starting point for modeling the tertiary structure of ZoPKS.
The HOMOLOGY and DISCOVER module (Insight II User Guide 1997, MSI San Diego, CA) of the Insight II molecular modeling package were used in the construction and optimisation of structures of ZoPKS and other selected PKSs. The computed models were verified for stereochemical quality by using PROCHECK at RCSB web site (http://rcsb-deposit.rutgers.edu). The graphical images of the models were viewed and generated by using Pymol graphical interface (http://pymol.sourceforge.net). The structural impact of amino acid substitution on the shape and size of substrate binding cavity of ZoPKS was investigated on a comparative basis. The amino acid residues forming the substrate binding region were mapped on all the models generated and the images were generated using the Pymol.
Gingerols, the major active components of ginger (Z. officinale), has received a great deal of attention because of its broad spectrum of biological activities including the anti-inflammatory, anticarcinogenic, and antitumor activities. Some initial investigations on gingerol biosynthesis identified the potential of enzymes of the PKS family as designers of its basic structural skeleton, but the lack of genomic information of PKS is a limiting factor to unravel the complex biosynthetic process. Degenerate oligo nucleotide primers targeting the conserved region of PKS was used for isolating the 600 bp core fragment of PKS by RT-PCR. This gene specific sequence was used for designing primers for the RACE experiments. The full length cDNA of PKS from Z. officinale (ZoPKS) was found to have 1173 bp length and was predicted to form a polypeptide of 43 kD containing
391 amino acid residues [Figure 1]. The BLAST analysis of ZoPKS gave a maximum identity of 64% to the PKS identified from W. thrysifolia. The low identity can be taken as an indication of its novel functions since the members exhibiting typical chalcone forming reactions are shown to have more than 85% identity(Brand et al., 2006).
The phylogenetic analysis of PKS superfamily showed clustering based on their biochemical functions [Figure 2]. The ZoPKS was found to cluster distinctly with the nonchalcone forming PKSs like the benzalacetone synthase from Rheum palmatum(Abe et al., 2001) , 2-pyrone synthase (2PS) from Gerbera hybrida (Eckermann et al., 1998), 4- coumaroyltriacetic acid lactone synthase (CTAS) from
Hydrangea macrophylla (Akiyama et al., 1999), valerophenone synthase (VS) from Humulus lupulus
(Paniego et al., 1999) and acridone synthase (ACS) fromRuta gravenolens (Springob et al. , 2000). The nonchalcone forming PKSs are involved in the biosynthetic processes other than typical tetraketide derivatives and because of their specific role, they are considered as speciesspecific
or metabolite speficic. Thus the characteristic clustering of ZoPKS may support for considering its role in nonchalcone forming reactions and possibility in the biosynthesis of pehnylbutanone derivatives of Z. officinale. The ZoPKS was cloned at the NdeI and BamHI site of pET32b for studying its expressivity in the prokaryotic system. The SDS-PAGE analysis showed the formation of recombinant protein of approximately 43 kD up on IPTG induction and was confirmed by the western blotting using the anti-His antibody [Figure 3].
Multiple sequence alignment of ZoPKS with the other PKSs showed that the conserved residues are located almost same position in all [Figure 4]. The sequence identity analysis of ZoPKS with the PKSs selected for the comparative analysis shows that it is identical to other PKSs selected for the analysis at a range of 49-64%. The maximum identity is towards the PKS from W. thrysifolia (64%) and the minimum is towards the A. arborescens pentaketide chromane synthase (49%). The multiple alignment analysis also revealed that the ZoPKS shows significant variations of catalytic residues at the highly conserved pockets. The catalytic triad forming the active site of the typical CHS like Cys 164, His 303 and Asn 336 are maintained as such in ZoPKS showing that it belongs to the PKS superfamily. The ZoPKS also showed the presence of stretches of conserved residues G(368)VL FGFGPGLT(378) considered as the signature sequence of PKS superfamily [Figure 1].
Structural insight into the catalytic machinery of polyketide biosynthesis was explained by the crystal structure studies of CHS from Medicago sativa (Ferrer et al., 1999). The studies showed that in addition to the catalytic triads, stringent conservation of amino acid residues constituting the catalytic pockets is also essential for the typical chalcone forming reactions. These include the substrate binding pocket (Ser133, Glu192, Thr194, Thr197 and Ser338) which forms a 16Å tunnel to the active site cavity and the cyclisation pocket (Thr132, Met137, Phe215, Ile254, Gly256,
Phe265 and Pro375). The overall architecture is maintained by the conserved amino acid residues like the Pro138, Gly163, Gly167, Leu214, Asp217, Gly262, Pro304, Gly305, Gly306, Gly335, Gly374, Pro375 and Gly376. A change in the amino acid residues at these pockets was found to have remarkable impact on the reaction mechanism leading to metabolic diversity (Abe et al., 2001). The ZoPKS showed significant amino acid substitution at these positions which makes it to be analysed in detail to unravel the effect of these changes.
Among the amino acid residues constituting the substrate binding pocket, ZoPKS showed variation from typical CHS by the substitution of Thr (197) to Ser and Ser (338) to Gln. Two amino acid substitutions are observed at the amino residues forming the cyclisation pocket by replacing Thr (132) with Ileu and Ileu (254) with Val. Furthermore, three amino acid residues are replaced at the strongly conserved residues shaping the geometry of the active site by changing Gly (306) to Asn, Leu (214) to Gly, and Gly (163) to Ala. The similar amino acid substitutions were observed in other members of the nonchalcone forming PKSs and these changes can be considered to have determining effect on the functionality of PKS. The amino acid substitution of S(338)I and T(197)L as observed in 2-Pyrone synthase when compared to the CHS was found to reduce the size of active site cavity to accept smaller substrates (Jez et al., 2000). Very interestingly, site directed mutagenesis studies revealed that substitution of just three amino acid residues (T197L / G256L / S338I) can make the CHS to have 2-PS activity showing the potential of key catalytic residues to regulate the reaction mechanism (Jez et al., 2002). Similarly, the PKS from W. thrysifola showed the replacement of T197 and Ser 338 with more bulky Met and Phe. This also have the characteristic Thr 132 to Gly substitution to provide the additional rotational freedom resulting in the size reducing effect (Brand et al., 2006). The active site analysis of Aleosine synthase showed a downward expanding T197A replacement, when compared to the typical CHS, illustrating the significance of Thr 197 as the controller of the polyketide chain length (Abe et al., 2006). Thus it is very probable that the amino acid substitutions observed in ZoPKS can alter the size and shape of cavity to favour the production of compounds other than the tetraketides. The structural impact of these substitutions on substrate binding pocket of ZoPKS was analysed in detail on a comparative basis.
Homology model of ZoPKS was generated based on the crystal structure of CHS (M. sativa) and 2-PS (G. hybrida) as templates. The energy minimisation of the modeled structure was carried out in the DISCOVER using the steepest descents algorithm (200 cycles) and conjugate gradient (1000 cycles) until a low energy derivative of around 1 kcal.mole-1 had been reached. At this point the calculated total energy of the molecule was consistent with the relaxed and stable conformation. The superposition of the ZoPKS model structure with the template structures showed very small deviation with an RMSD less than 0.5 Å and 0.8 Å respectively respectively towards the 2-PS and CHS [Figure 5]. Similarly the homology models were also generated for PKSs selected for the comparative structural analysis. The quality of all the models was assessed by Ramachandran plot analysis using PROCHECK. It helps to check the stereochemical quality of the optimized structures. By the inspection of phi/psi angle, we can analyse the quality of the Ramachandran plot and can determine whether the protein conformation belongs to the allowed region or not. The Ramachandran plot analysis of the ZoPKS showed that 92.9% of the amino acid residues come under the most favorable region, 6.2% amino acid residues come under the additional allowed region and 0.6% amino acid residues come under the generously allowed region. The same anaysis for the crystal structure of CHS (M. sativa) and 2-PS (G. hybrida) gave the result of 93.4% and 92.6% amino acid residues at the most favorable region which strongly support the stereochemical quality of the model. The Ramachandran plot analysis of other PKSs selected for the study also showed the distribution of 91-93% of amino acid residues in the most favourable region [Table 1].
Although the PKSs have promiscuous substrate specificity, the amino acid residues constituting the substrate binding region was found to have mechanistic and steric effects on the substrate selectivity (Brand et al., 2006). This is also supported by the results of the multiple sequence alignment in which the conserved residues forming the substrate binding region were maintained as such in the CHSs from Arabidopsis thaliana, Iris germanica, Oryza sativa and Dendrobium nobile. But characteristically the amino acid residues were substituted in other PKSs specifically for their specific functions. So these residues were selected for the comparative structural analysis of ZoPKS for making predictions about its novel functions possibly as a nonchalcone forming PKSs. The RMSD values of the Cα atoms of all the amino acid residues forming the substrate binding pocket like the Ser 133, Glu 192, Thr 194, Thr 197and Ser 338 were calculated for the homology models by overlay of the Cα atoms of generated models with the CHS of M. sativa. A number of substantial differences were observed for the RSMD values of ZoPKS and these differences along with the changes in the positioning of the side chain indicate the distinct structural characters of ZoPKS [Table2]. Very interestingly the amino acid residues at the 338 position of ZoPKS showed an RMSD value of 1.76 ? where the Ser is substituted by more bulky Glu. The Ser at the 338 position was found to have role in starter substrate selectivity and is more prone to substitution in members of nonchalcone forming PKSs for the specific function (Abe et al., 2006). In order to understand the combined effect of amino acid substitutions at these well studied positions, the substrate binding residues were mapped on modelled structures of ZoPKS and other PKSs selected for the comparative analysis. A significant variation is observed at the substrate binding region of PKSs and the changes observed in ZoPKS supports its novel functions [Figure 6]. The proposed reaction mechanism for the gingerol biosynthesis demands the PKS to have altered substrate selectivity, altered condensation and cyclisation reactions when compared to the typical CHSs. So the amino acid substitutions observed for the ZoPKS may contribute the changes in the structural architecture of ZoPKS to perform this novel function. But it has to be demonstrated by in vitro studies to confirm and the current study forms a genomic basis for this. As the molecular fascination of PKSs is just begin to explore, the remarkable changes observed on the PKS identified in the study show that it forms a potential member of PKS superfamily.


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text:
"Well," Philomel said after his harmless moment of laughter. The sounds of his merry chortling still rung about her ears. Strangely, it was a pleasant sound, and neother mocking nor irritating. In fact she seemed to grasp some hnour and pride out of the fact he found her funny - even though her egoism caused her not to comment on this fact, or even show it in her expression.
"Well," she repeated, a couple of seconds later, for the effect of not seeming easily impressed. "Shall we continue on our way?"
Breaker paused and looked rather pointedly at her unconcious beast of burden. Pursing her lips, Philomel had to confess he had a point, and so simply nodded, and sat right down where she was.
"We shall go when he wakes then?" she suggested, but really ordered. After all, her furry rear-end was already on the soft earth.
The maul-slayer inclined his head and agreed, taking his time to sit opposite her. It was done in a graceful way, and the faun could not help but be impressed by this god-favoured being. Certainly, he had the air of glory about him, a feeling of strength diginity. In her pleased heart she quietly contemplated him, his figure and demaneour, quite taken by him - but by no means taken aback. Rather, it was a joyous thrill to watch him - watch the bend of the knightly knee, the exquisite fold of the nimble hand. As her lips pursed and the faun lost herself in the measure of percieving him, Veridian chose the time to sidle up beside her.
Yes, he is very pretty, he muttered, somewhat in a sickened tone, into her head.
Softly, she blinked, and pushed away his pointed muzzle from where it gestured to her. Meanwhile did she never take her eyes off Breaker's seated form.
You can be quiet now, replied she, but in a high-strung, non-chalant tone.
Veridian snickered away. Brushing his tail against her knee he moved his head away from her, choosing instead to curl up near her hoof. In this position he was between her and the mighty horned head of the sleeping tera'k, purposeful to come to guard when the animal awoke. Likely to hefty bellows and copious thrashes.
Not too far to the north, hidden away in a chamber beneath the forest's darkness, there sat a man and a woman. Her face was half hidden in shadow, for the fabric of a cowl was thrown over it. He himself had little to hide his identity besides a mass of wild hair framing a strong-jawed face. Whilst she was thumbing the ridge of a maple elf-made bow, he dug the end of the shaft of a mattock into the ground. Across from each other they were positioned, between them nothing but a space of earth and small stones.
"So," said he in a very low, husky, but booming voice. "We are 'ere."
His accent, an urban backstreet one by the best of guesses, rolled into the gloom of the cave and tickled the curving, upwards entrance. A few mindless mites were sent scattering by the volume.
It caused the female archer to arch an eyebrow before she replied.
"Indeed we are," her voice was far more proper. Born and raised in a noble house it only made sense. "We finally meet."
"I am Feardon."
"And I am No-One," she quickly responded, the bright flashes beneath her hood that were eyes glaring at the man. "And that is what I am, and that is what you will know me as."
For a moment the man was tempted to burst into laughter, but something of the way she stared, and something of the way she fingered her bow and the nearness of her full quiver told her it would be a mistake. Pride placed aside, he inclined his head.
"Fine, then No-One ye'are, and no one ye shall be. 'Tis a pleasure to meet ye."
"And you too, I suppose," No-One raised her chin slightly, using her spare hand to scratch underneath it, eyes though never leaving him. "There is a lot I have been told, and a lot I am expecting."
Feardon smiled a small bit - or perhaps it was more of a smirk. His massive hands ran down the shaft of his mattock and hit the cold steel edge of the metal heads.
"Well it takes a lo' to be in service to a demon-god," he said in a sort of testingly idle way. At the sound of his frustratingly loud, irating tone No-one siezed, and it took more willpower for Feardon not to burst into laughter. Quickly, he continued his sentence to avoid her wrath. "... So I am guessin' you are rather expectan' of many people. Tell me, 'ave you ever worked with a professional barbarian before?"
"No, but I have killed your kind before," she said with no hesitation, "And I will do again, if you get in my way."
"But-" pointed out he, fast, before she could even begin to be tempted, "If ye do, then you will be alone in this. And the reports say there are two of them searchin' fer the lyre."
"The lyre which we will get to before," No-One said, her chin jutting up in pride. "Which we will take and track before they do."
"To use fer your god's purposes I suppose?"
Her head bobbed once, a decline and an incline to signify her acception of the question. "All for his purposes. As soon as my temple received word the whereabouts of Orphaeo's tomb has possibly been discovered, we knew we had to act. I was sent here, to work with you, my hired help, in order to get it before any others do."
"I am yer hired help am I?" Feardon raised a brow.
No-One pointed in a direct manner to the large, heavy pouch at his belt. The barbarian chortled somewhat, finally for a legitimate reason, as he cupped the clinking bag of coin with a hand.
"I guess I am," his grin spread across his magnificent, scarred face, "I guess I am then."


text:
3 Women- Two Women In Museum With Painting Nana By
painting by artist Gerard Boersma
6 in X 6 in (15.2 cm X 15.2 cm)
acrylic on masonite
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text:
Baseball Opens 2016 Home Schedule on Wednesday
LEXINGTON, Ky. -- After a six-game road swing to open the year, the Kentucky baseball team will open its home schedule with the first of a season-long 10-game homestand on Wednesday, welcoming Austin Peay for a 4 p.m. ET first pitch at Cliff Hagan Stadium.
The game will be broadcast live on SEC Network + and will be available through the UK Radio Network, locally on 1580-AM and online at UKathletics.com.
Last weekend, Kentucky opened its 2016 season with six consecutive games away from Lexington, including two wins on opening weekend in Spartanburg, S.C., before traveling to the Tony Gwynn Classic. UK fell in the opener to eventual champion UC Santa Barbara, before winning at San Diego State on Saturday and then falling in the finale to preseason Northeast Conference favorite Bryant on Sunday.
On Friday, UK fell behind due to big innings in an 11-4 loss to UC Santa Barbara. The Wildcats then bounced back behind a strong starting pitching performance from senior Dustin Beggs on Saturday. Beggs, named the Gwynn Classic Most Outstanding Pitcher, worked eight-shutout, two-hit innings, striking out 10. UK got a three-run homer from Riley Mahan to power the win. Sunday, Bryant plated seven runs in the first five innings for a six-run lead, with UK's late-inning rallies thwarted by key inning-ending double plays in the seventh and eighth.
Kentucky has hit .302/.375/.547 through six games in 2016, with 13 doubles, three triples, 11 homers and 41 runs scored, stealing four bases. On the mound, UK has a 4.15 ERA as a team, striking out 54 and walking 17 in 52 innings.
Individually, preseason All-America third baseman JaVon Shelby has led the team with a .458 (11-for-24) clip, with five doubles, two homers and four RBI. Mahan has a .450 (9-for-20) average with two doubles, two triples, one homer and four RBI. Second baseman Luke Becker has hit. 368 (7-for-19) with a double and two RBI, and outfielder Zach Reks sports a .316 (6-for-19) average with two RBI. Evan White has hit .286 with three doubles and four RBI, with Tristan Pompey leading the team with three homers and eight RBI, batting .286 with a double and triple.
On the mound, UK has used the weekend rotation of junior Zack Brown (0-2, 9.00 ERA) on Friday, senior righty Dustin Beggs (2-0, 0.64 ERA) on Saturday and senior righty Kyle Cody (1-1, 2.61 ERA) on Sunday. In relief, Zach Logue (0-0, 0.00 ERA), Sean Hjelle (0-0, 0.00 ERA), Zach Strecker (0-0, 1.69 ERA), Josh Smith (0-0, 9.00 ERA), Logan Salow (0-0, 10.12 ERA) and Brad Scaenzer (0-0, 27.00 ERA) have each appeared in two games.
Austin Peay (4-3) has hit .321/.389/.554 as a team through seven games, with 22 doubles, five triples, eight homers and 55 runs scored, stealing eight bases. Defensively, Austin Peay has allowed seven steals, fielded .969 and turned three double plays. On the mound, Austin Peay has a 6.57 ERA as a team working in 63 innings, allowing 71 hits and 24 walks, striking out 58 and holding opponents to a .278 average.
Individually, Dre Gleason leads the Gov's offense, hitting .538 (14-for-26) with four doubles, two homers and 15 RBI. In relief, Brett Newberg (0-0, 0.00 ERA) has appeared in four games, along with Jordan Harmon (0-0, 15.00 ERA).
Kentucky leads the all-time series with Austin Peay, 11-2, winning the last five meetings, most recently a 6-3 win in 2013 in Lexington. UK leads the series in Lexington, 9-1.
Kentucky: 3-3
Austin Peay: 4-3
Schedule (times Eastern and subject to change) -- Lexington, Ky.
Wednesday, March 2 * vs. Austin Peay * 4 p.m.
Television (SEC Network+)
SEC Network+ and WatchESPN
Radio (UK Radio Network)
Wednesday: Fox Sports 630-AM in Lexington
Internet Coverage
Live audio and stats on UKathletics.com
Tentative Pitching Matchup
Wednesday vs. Austin Peay
TBD vs. TBD
Cliff Hagan Stadium (3,000)
Free gameday parking is available in the C-8 lot beyond rightfield and behind homeplate, or in the E lot adjacent to Cliff Hagan Stadium. The C-8 lot will be filled on a first come, first served basis. Once C-8 is filled, vehicles will be directed to park in the E lot next to Cliff Hagan Stadium. After the E lot is full, a limited amount of parking will be available in Parking Structure No. 7 on the ground floor. If you're parking in PS#7, please exchange your ticket for a pink payment voucher at the ticket booth, prior to the end of the fourth inning. Additionally, overflow parking will be available in the smaller E lots close to Cooper Dr., as well as the C-6 lot at Commonwealth Stadium. All fans are asked to please follow the signage provided to the nearest parking location.
$5 - General Admission Seating - Adults
$2 - General Admission Seating - Kids/Seniors
$35 General Admission
$55 Reserved Seating


text:
chemistry (work shown)
posted by tatiana .
How would you make a serial dilution if you have a starting value 100mM of X
The are the values of X show different dilutions.
Volume variables (x) mL
We starting out with 100mM of X which would be considered its concentration. In order to describe how to do a serial dilution I need to figure out the dilution factor from the volume variables. She wants us to describe how when starting from 100mM of concentration we can obtain those volumes.
Df1= 100ml/30mL = 3.333
Df2= 30mL/ 10= 3
Df3= 10mL/3= 3.333
Df4= 3/1 = 3
Df5= 3/1= 3
so I guess she wants us to work our way from the bottom up so I obtained a dilution factor of 3.333 and 3.
to find out the concentration I have the formula
C= C1/Dfactor
C= 100mM/ 3.333= 30.03mM
how would we describe doing this dilution. From the 30.0mM concentration we add 3.3333 mol per liter. As you see I am still confused. I have no way of asking her for explaination, so If any help could be proivded I would appreciate it.


text:
end of May
Last weekend was outstanding. Many I talked to were catching fish. I was out and caught 4 rainbows 18-20" in the AM near Zoar Gap and down to Pelham Brook. They were not rising when I was there and seemed to like the basics-got 'em all on a dark Wooly Bugger. Next day was a little slower, I missed one and landed one but still nice out there.
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text:
Thinktiv Playbook: Amplifier
Prysm is a leading provider of video wall solutions, based on the company's groundbreaking Laser Phosphor Display (LPD) technology. Backed by nearly $150M of private equity capital, Prysm engaged Thinktiv as their outsourced, turnkey partner for visual brand design, market messaging, marketing content development, and demand generation programs. Thinktiv played this critical role for the company for over four years, supporting the expansion of Prysm's product line and initiatives into new market sectors.
Services: Market Analysis + Branding + Messaging + Website Design + Content/Collateral + Go-to-Market Strategy + Multi-channel Marketing Programs
Market Sector(s): Video Display, Hardware, B2B
With a world-class design and demand generation team on call, anything is possible.


text:
Had the pleasure of enjoying the tUnE-yArDs live yesterday at the 5TH ANNUAL ROOTS PICNIC. They didn't disappoint. Here's a taste in collaboration with ?uestlove, Angélique Kidjo and Akua Naru for Red Hot + FELA — a new Fela Kuti compilation by the Red Hot Organization and the (RED) campaign.
As always, you're welcome.


text:
Audio Tools and audio editor
Use this security logo:
Tutorial not found. Let know us about any useful video tutorial.
audio converter, audio editor, audio tool, vocal remover, mp3 key shifter
Karino Audio Tools (english version)
You are hereby licensed to make as many copies of the shareware version of surfmaster and documentation as you wish; give exact copies of the original version to anyone; and distribute the software and documentation in its unmodified form via electronic means. There is no charge for any of the above.
Questo software e' SHAREWARE. Alcune funzioni sono disabilitate nella versione di prova. Per avere tutte le funzionalita' devi acquistare la licenza.


text:
On Wednesday, three hyenas shall be slain, no matter what the outcome of the vote will be. The first to be put out of contention will be fear.
Barely two and a half years ago, lives were lost, property destroyed and relationships poisoned. For many Kenyans at that time, one's ethnic background was all that was required to put one in harm's way. Fear stalked the land.
The post-election violence went further than that; it raised deep concern about the future viability of this nation. We had slipped so far towards dismembering the country that it was difficult to see how we could again live harmoniously as a nation.
With the referendum vote, we will be saying that we chose to live together, guided by a constitution we all want to uphold, be it the new or the familiar one.
No longer shall we have to put up with a lurking, numbing fear of ourselves and what we can each do to each other. The vote will also be a rendezvous with the hyena of violent dispute.
This time we will be reaffirming that matters national can, and should, be decided through a peaceful, free and fair voting process.
We will be acknowledging that even when we hold different opinions, an electoral process free of malpractices shall be sufficient to settle the matter.
The referendum will present an opportunity to cast off the stigma of adulterated elections. It is a chance to forever forsake bloodshed as a political tool for settling electoral contests.
With peaceful voting, we will be declaring that we shall neither soak our lives with blood nor stain our land with senseless death and disgrace.
We will be proclaiming, almost half a century since we gained independence, that we are a mature enough nation to hold a credible vote that will effectively settle our debates and disputes.
The third hyena to be vanquished will be that of tribal politics.
With choice phrases based on stereotyped prejudice, politicians have been able to whip up hordes of virulent supporters.
Even though campaigns on the proposed constitution have employed aspects of tribal regimentation, our politics has, on the whole, become much "cleaner"� over the past few months. It is certainly healthier than the last referendum which polarised the country along tribal lines.
In contrast, cleavages on the proposed constitution cut across ethnic and generational gaps, bringing us the closest we have been to an issue-driven campaign.
Significantly, new and passionate interest groups that are based on common perspectives about what is best for this nation are emerging. At the heart of such groups are concerns that transcend tribal issues and divisions.
Nonetheless, by Wednesday, some citizens will, sadly, be looking to their tribal chiefs for guidance on how to vote.
However, many will be taking a fairly tribe-neutral stand and will cast their ballot based on their judgement of the issues.
In short, on Wednesday, we will be stating that an electoral process is not a proxy war to be prosecuted by "warlords"� leading excitable tribal regiments that overlook the merits of the issues.
We will demonstrate that elections and referenda can be peaceful so that eventually, the most popular options carry the day.
Mr Muleji is a consultant with Upward Bound. (firstname.lastname@example.com).
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text:
Nick and Vanessa Lachey: Farewell L.A.!
Taking a family trip out away from L.A., Nick Lachey, Vanessa Minnillo, and their son arrived at LAX Airport on Thursday (April 25).
The former "Wipeout" host was dressed casually in a white top, blue jeans, and white sandals as she pushed baby Camden's stroller behind her hunky hubby.
Though currently living in California, the 98 Degrees frontman recently revealed their plans to raise Camden in Ohio.
Speaking with "Access Hollywood Live," Nick shared, "I really value the effect that family has on a child growing up. I want Camden and other future kids we have to be around their cousins and their grandparents. That's a very important part of growing up."


text:
sebacod3 (05-05-2012)
Just found about that side,
Log in with your Origin account and you will get instant acsess to the M1911/TacLight
Battlefield 3 Weapon unlock
Have fun
Last edited by fatjoe2015; 05-03-2012 at 03:07 AM.
sebacod3 (05-05-2012)
Can some1 tell if this works?
Thanks me if i help you :P
OFC it works... i did it when game was out. It is totally legit
Nice post, will help some people I'm sure.


text:
For a crash course in health and safety, it's best to turn to the experts.
Little League baseball players, parents, community members and Hall County District 2 Commissioner Billy Powell were on hand Monday night as the words they had longed to hear were finally spoken: "Play ball."
Not all competition has to be related to sports or athletics, as many college students found on Saturday at the University of North Georgia Gainesville campus.
Spring break is almost over for the Gainesville and Hall County community, and with it ending, local blood banks are hoping to see a rise in blood donations.
Saturday was a perfect day in more than just weather for those who like to help others, give back or just be part of a church that serves its community.
ATHENS - U.S. Army recruits got a glimpse Saturday into what their military lives will be like during a pre-basic and physical training at the U.S. Army Reserve station off Winterville Road in Athens.
Gloria Stargel, a local Christian author and inspiration to people throughout the Gainesville community, died on Saturday.
If children like sports, they can usually join a team. However, if children like math, they don't have an outlet for competition.
The Exit 17 welcome sign for Oakwood off Interstate 985 is back up after being blown down by high winds on Dec. 20.
Sometimes a risk never pays off with a reward. A selfless act may only get a "thank you."
The Three Sisters Vineyards in Lumpkin County announced it will be the new location of an official University of Georgia Extension and Georgia Automated Environmental Monitoring Network weather station.
Former NFL player and coach Dan Reeves, spoke to the cadets at Riverside Military Academy about character development and leadership on Wednesday morning.
The Vietnamese congregation of First Baptist Church rang in the new year Sunday afternoon at the church's banquet hall in Gainesville.
CLEVELAND - The North Georgia Zoo and Farm has a new addition - a baby albino wallaby named Joey.
The Gainesville High School Red Elephants football team was honored and celebrated Wednesday afternoon with a parade and reception for its Class AAAAA state championship victory over Ware County last December.
Page 10 of 14
Privacy policy and Terms of service


text: Trying to decide whether or not I can afford W.W. Chan, but they have no prices listed on their website. I realize that price can vary greatly depending on fabric and details, but can any Chan clients here give me an idea of the likely cost of a basic sport coat? Thanks.


text:
Can software only be scanned not entered in manually
This topic was created during version 1.7.
The latest version is 7.5.00101.
1 Reply
· · ·
Oct 8, 2007 at 12:00 UTC
Right, it is currently scanned not entered manually. You could use user-defined assets to track it, but it doesn't allow you to enter the kind of information that the software section contains. You'll probably want to check out the feature requests forum and add you vote to the applicable threads there.
This discussion has been inactive for over a year.
You may get a better answer to your question by starting a new discussion.


text:
Hilton Northampton Hotel
100 Watering Lane, Collingtree, Northampton, NN4 0XW, United Kingdom – Show map
|Room Type
Deluxe Double Room
Double Double Guestroom
Double Hilton Guestroom
Deluxe Double/Double Room
Twin Hilton Guestroom
5 Reasons to Choose Hilton Northampton Hotel
Prices you can't beat!
Manage your bookings online
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Hilton Northampton, a business and family-friendly hotel, is easy to find off the M1 and only 40 minutes from London Luton Airport (LTN). We have ample function space in our 12 meeting rooms, a health club with heated pool and beauty treatments, and a restaurant offering a kids' menu. Relax by the pond in our courtyard before heading out to exciting attractions nearby.
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What would you like to know?No thanks
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Facilities of Hilton Northampton Hotel Great facilities! Review score, 8.1
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Until 12:00 PM
Cancellation and prepayment policies vary according to room type. Please enter the dates of your stay and check what conditions apply to your preferred room.
Children and Extra Beds
All children are welcome.
There is no capacity for extra beds in the room.
Pets are allowed. Charges may apply.
Cards accepted at this property
Hover over the cards for more info.
When booking for 10 rooms or more, different policies and additional supplements will apply.
A security deposit of GBP 50 per night is required on arrival. This deposit is fully refundable upon check-out and subject to a damage inspection of the accommodation.
Please note, children 16 years and younger must be accompanied by a parent/guardian in the hot tub, swimming pool and leisure facilities. An adult guest can supervise a maximum of 2 children aged 16 years and younger at any one time.


text:
Jimmy Kimmel: Little Known Facts About the Late Night Show Host
Jimmy Kimmel, host of the ABC late-night show bearing his name, is widely recognized for his wit. But you don't know everything about him.
For instance, Kimmel unabashedly takes credit for having the world's record for creating the Biggest Meatball – at some 270 pounds, or the shape of a basketball on steroids.
You also might not register that Kimmel is one of the most forward-thinking people in show business in the way he employs the Internet as part of his marketing plan. Kimmel has used the Web to popularize such well known videos, based on sketches from his show, as "The Handsome Men's Club" and I F***** Ben Affleck," with whom, Kimmel cheerfully reports, is "still hot and heavy."


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