By Forrest Milburn, Staff Writer July 24, 2017 Updated: July 25, 2017 12:12 a.m.
Serving as the gateway to South Texas and the state's Hill Country, San Antonio has a rich, diverse history shaped by a large population of residents with Mexican ancestry dating back before the Texas Revolution.
This history is reflected in the fact that one of the city's early mayors was of Mexican descent. But it took more than a century for San Antonio to elect its next Latino mayor. In recent years, the city has had a handful of mayors blaze the trail for historically disenfranchised groups.
"With a majority-minority population in a very conservative state, that says a lot about the diverse population in our city willing to elect people from other backgrounds," she said.
The city, now the seventh largest in the country and the second largest in Texas, has a population of nearly 1.5 million, with a demographic breakdown of 73 percent white, 7 percent black, and 2 percent Asian, according to the U.S. Census Bureau's 2010 survey data. When factoring in ethnicity, the city is 63 percent Hispanic or Latino of any race.
But even with such a large Latino population, the number of elected mayors from that community can be counted on just one hand.
The city's first mayor of Mexican descent since Texas' independence may sound familiar.
Juan Seguin, whose last name can be found not just on maps showing the city east-northeast of San Antonio but on roads and buildings throughout the metro area, served as mayor from 1834 to 1835 and again in 1841 after the state's founding as a republic, in which he played an instrumental role in serving as a military leader in the Texas Revolution.
Seguin, who was also the first Tejano to serve in the Republic's Senate, oversaw the city during a time of great tension between Anglos and Mexican Texans. He resigned in April 1842, fleeing for his life to Mexico when he was accused of aiding the Mexican military.
Nearly a century and a half later, in 1981, Henry Cisneros, the youngest City Council member in San Antonio history, became the first Latino to serve as mayor of a major U.S. city during the 20th century. He was re-elected three times, ending his tenure in 1989.
"He was the first 'modern' Latino mayor," Navarro said. "It was pretty much an all-white City Council and a majority white community," and by being politically savvy, he was able to overcome several barriers to get into elected office, she added.
Cisneros succeeded by building the right coalitions and having a progressive vision needed to win elections. The path he took is one any other Latino or member of a minority community should replicate in running for mayor, Navarro said.
After serving as mayor for two terms, Cisneros took his expertise in city planning to the U.S. Department of Housing and Urban Development in the Clinton administration, where he served as secretary from 1993 until 1997.
"He's still a very important role model for the Latino community here in San Antonio," Navarro said.
Mayor Julián Castro, only the third modern day Mexican-American mayor, followed in Cisneros' footsteps by leaving City Hall to serve as HUD secretary in the Obama administration in 2014. Ed Garza was mayor from 2001-2005.
In the void left by Castro's resignation, the City Council unanimously selected Councilwoman Ivy Taylor as the city's next mayor. Taylor, who went on to run for the seat in 2015 and for re-election in 2017, became San Antonio's first African-American mayor.
At the time, Taylor's selection as mayor made San Antonio the largest city in the U.S. to have an African-American woman in power.
"It certainly was cutting edge at that time," said Jim Brooks of the U.S. Center for City Solutions, an urban research arm of the National League of Cities.
But after Taylor was first sworn-in, she tended to downplay her historic election, instead focusing on her plans for the city rather than herself.
Taylor, who was recently defeated in the mayoral election by Ron Nirenberg, was also only the second woman to hold the mayorship in San Antonio history.
The title of the city's first female mayor belongs to Lila Cockrell, who served in the office for four two-year terms after sitting on the City Council for almost a decade.
First elected in 1975, Cockrell led San Antonio through a time of great economic and population growth, helping bring the HemisFair to the city in 1968. She also helped guide the City Council from at-large representation to single-member districts.
"(At the time,) we were a very macho, very male-centered city with a male-centered city council," Navarro said. For her to be able to overcome the political landscape in winning "says a lot about breaking that glass ceiling," she added.
Cockrell served as mayor from 1975 to 1981, when she left office to take care of her ill husband. Then, after Cisneros decided against running for re-election in 1989, she was persuaded to run for a fourth term and won. She was defeated in 1991 for a fifth term when she placed third behind two City Council members.
Since her tenure, there have been several unsuccessful attempts by a few Latinas to run for mayor, including Leticia Van de Putte, a former state senator, who ran against Taylor in 2015. Their run-off election was the first for the city with two women of color.
Nirenberg, the city's current mayor, is the first in the city to have Asian roots, with Malaysian ancestry on his mother's side and Russian and Polish ancestry on his father's side.
So what's next in San Antonio history?
Navarro said the city has yet to have a Muslim or a member of the LGBTQ community as mayor, but it's not inconceivable.
COMING WEDNESDAY: East French Place proposed as historic district.


About Us - Nightingale Electrical Ltd.
Nightingale Electrical has grown from the basement of founder Doug Nightingale's house to an 8,000 square foot office in just over 30 years. We've grown steadily over the years by taking advantage of common-sense business practices, diversified capabilities, and an unwavering dedication to high quality work.
This attitude has allowed us to build a staff of over 200 and maintain long-term relationships with the most well respected development companies in BC; organizations like Onni, Bird Construction, Ledcor, Ventana, and Adera Properties. Thanks to these partnerships, we've been able to work on some of the most noteworthy construction projects in Greater Vancouver over the last two decades; from wood frame condominiums to projects like the Living Shangri-La, the Bayshore Gardens Towers, and the Ballard Power Systems office and plant.
– Doug Nightingale, Founder and CEO.


TONIGHT'S MATCHUP: Tonight is the seventh meeting between Adirondack and Manchester this season, with the Thunder holding an edge in the series thus far, earning points in five of the previous six matchups with a 4-1-1 record. The most recent matchup between the two teams came back on December 27th, when the Monarchs shut out the Thunder 2-0. Adirondack comes into tonight's game off a 3-2 victory over the Reading Royals in Pennsylvania on Sunday, while Manchester comes in fresh off of back-to-back shootout loses to the Greenville Swamp Rabbits at home.
COMIC CONS: With his goal on Sunday afternoon, Thunder defenseman Ryan Constant added another point to his impressive power play totals. The veteran defenseman has 17 total points (3-14-17) on the season with nine (2-7-9) coming on the man advantage, tops on the team. The ninth year pro has already tied his season total points from last season, and is on pace to break his career high for points of 36.
SWEET HOME GLENS FALLS: Wednesday's matchup is the first of a five-game homestand for the Thunder, where the team will face off against Manchester, Reading and Norfolk individually, while also playing back-to-back games this weekend against the South Carolina Stingrays. The team enjoyed great success at the Civic Center in 2015, posting an 8-3-1 record on home ice.
CLIFTON PARK CONNECTION: Two local skaters, defenseman Patch Alber and forward Peter MacArthur, made important contributions over the past week. Alber totaled three assists, including two in Sunday's win, which led the team en route to being named the Thermal Associates Player of the Week. MacArthur scored the lone goal in Thursday's win and added the shootout winner to boot.
STEPPENWOLFE: Since he returned to the Thunder on December 18th, forward Greg Wolfe has stepped up and notched five points (3-2-5) in seven games, including the game winning goal on Sunday in Reading. Through 21 games with the Thunder this season, the Canton, MI native has amassed 15 points (6-9-15), which is fifth best on the team.
SIMPSON PLAN: The Stockton Heat assigned goaltender Kent Simpson to Adirondack yesterday afternoon. The Edmonton, ALTA native has appeared in two games this season with the Thunder, where he went 2-0 with a 2.00 goals against average and a .939 save percentage. Simpson, who also appeared in seven contests this season in the AHL with the Heat, gives the Thunder a three-headed monster in goal, combined with Ken Appleby and Drew Fielding. The three netminders have combined for a 16-8 record this season.


Bioniq is a family oriented Internet Service Provider located in Fourways, Gauteng. We started the company in August 2015 as a wireless company with a vision to bring a true high-speed Internet solution to the market, one that would keep on pushing the boundaries of internet connectivity. Till this day we have grown exponentially into a fibre service provider while staying true to the mission and core values the company was started on.
Our mission is to surpass the needs and expectations of our customers by offering the fastest and most reliable internet solutions available in South Africa at the most competitive prices. We do so by staying up-to-date with the latest technological advancements in the industry and by being the first to deploy those technologies across our networks. We identify our customers needs early-on and foster early adoption of new advanced technologies. By providing leading-edge innovation, we connect our customers to every corner of the world empowering them to create their own paths to success and to improve their lives and the lives of those they serve.
Our vision is that we will be one of South Africa's Leading Internet Service Providers of Choice offering our customers a wide variety of the most advanced and effective solutions readily available in the market. We aim to be a industry leader known for our innovations, early adoption, reliability, ethics, professionalism and all round unique and superior customer experiences.
Winning – We never settle for second best, we always deliver high quality work and strive to exceed expectations.
Passion – We are committed in heart and mind and we love what we do.
Collaboration – We collaborate in all aspects of our business, we work as a team to take advantage of our collective genius so that we succeed together.
Integrity – We are open and totally honest: our business is ethically and morally strong and each of us is accountable.
Innovation – We strive to be ahead of the game by continually improving the way we do things for ourselves and our clients.
Bioniq seeks strategic partnerships to create synergies that will enable us to achieve more than we could alone. By partnering with us, we will bring together organizations and individuals to eliminate boundaries and create value for those involved.
To discuss a potential partnership send us a mail via the contact us form.
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In an tweet issued from the team's accountit offered a glimpse of what seems like a fresh front wing decorated with a pattern.
If the brand new colours extend beyond the front wing, or Even though the team has not elaborated on its plans for its W10, Tweet and the layout could be a clue from exactly what it's streak 48, that Mercedes is currently opting for some thing this season.
Mercedes is due to unveil its fresh W10 on February 1-3, before the earliest official evaluation taking place at Barcelona in February 18. However, the team might not run the livery in analyzing as it appears at the launching.
With the blueprint reminiscent of a design livery that road car manufacturers use to disguise the design theories, it could be that Mercedes is intending to cover its work. Red Bull famously conducted the camouflage livery on its car.
The colours might at least make it harder for teams to grab and copy some fine details of this Mercedes, with teams having devoted lots of resources to optimizing front and rear wings.


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Seth Grob
Mr. Grob has taught and frequently lectures throughout the country on adoption, infertility law and child welfare issues. A list of a few of his numerous presentations is listed below. He has also served on the faculty at the University of Denver College of Law and Red Rock Rocks Community College where he has taught classes on adoption, juvenile law, child advocacy, and child welfare.
List of Some Presentations:
Duke University Law School, Silent Victims: Foster Care and Foster Care Adoption in America: ("The Opioid Crisis: What Can Be Done for the Children"), (Durham NC, November 2018).
Adoption Options, "Expedited Relinquishments and Contested Adoptions," (Denver, July 2018).
American Academy of Adoption Attorneys Annual Conference, "Subsidy & SSI Benefits for Special Needs Children: Part II," (Montreal, Canada 2017).
1st Judicial District Judges and Magistrates, "Contested Adoption Cases and Best Practices in Relinquishment Cases," (Golden 2015).
ABC News, "Interview with Seth Grob Regarding the Cryopreserved Embryo Dispute involving Sofia Vergaras," (2015) [The interview can be seen on YouTube].
Rocky Mountain Immigrant Advocacy Network Conference, "The Intersection of Family Law and Immigration Law," (Denver 2014).
American Academy of Adoption Attorneys Conference, "Meeting the Needs of Older Children Who Lack Immigration Status and/or Who are in Foster Care," (Philadelphia 2014).
Colorado Children's Caucus, "Failed Adoptions: Policy Issues Surrounding 'Re-Homing' and the Need to Keep Children Safe," (Denver 2014).
Office of the Child Representative Summer Conference, "Adoption: Considerations for Guardians ad Litem," (Silverthorne 2013).
American Academy of Adoption Attorneys Annual Conference, "Adopting From the Foster Care System," (San Diego, April 2013).
American Academy of Adoption Attorneys National Conference, "Assisted Reproductive Technology Case Law and Legislative Update," (Indianapolis 2012).
Creating a Family Radio Show, "Adoption Subsidies for Foster Care Adoptions: What is Reasonable and How to Negotiate," (November 30, 2011).
Colorado Office of the Child's Representative, "The Role of the Guardian ad litem in Adoption Subsidy Cases," (Denver 2011).
North American Council on Adoptable Children's 37th Annual Conference on Healing Across Systems, "Dissolved and Disrupted Adoptions: Adoption Subsides and Other International Issues," (Denver 2011).
American Academy of Adoption Attorneys Mid-Year Conference, Strategies for Adoption Agencies and Attorneys: Loss Prevention, Management, and Litigation Covering Disrupted and Wrongful Adoptions, "Adoption Subsidies and International Issues in Disrupted Adoptions," (Minnesota 2010).
American Academy of Adoption Attorneys, "Disrupted Adoptions," (Telephone Seminar 2010).
Family Law Institute, Domestic Relations Law in Times of Economic and Family Crisis, "Navigating the Interstate Compact for the Placement of Children," (Breckenridge 2009).
American Academy of Adoption Attorneys National Conference, "Adoption and Kinship Guardian Assistance, New Developments in Federal Law," (Portland 2009).
American Academy of Adoption Attorneys National Conference, Gateway to Adoption, "Help Us: Advocating for Adoptive Parents and Waiting Children," (St. Louis 2008).
Lutheran Adoption Network National Conference, "Securing Adoption Subsidies for Special Needs Children and an Update on the Proposed New Interstate Compact for the Placement of Children," (Denver 2007).
Florida Adoption Council Statewide Conference, "Adoption…What Every Practitioner Needs to Know," (St. Petersburg 2007).
National Business Institute, Adoption Law: Start to Finish, "Overcoming Intrastate Adoption Issues," (Denver 2006).
American Academy of Adoption Attorneys Mid-Year Conference, Agency Adoptions and Foster Care: Emerging Legal Issues for Attorneys, Social Workers, Foster Parents, and Other Adoption Professionals, "Advanced Adoption Subsidy Issues," (Denver 2006).
Resolve of Colorado Statewide Conference, "Legal Issues in Adoption," (Denver 2005).
Colorado Office of the Child Representative. Best Efforts and Practices in Pediatric Law, "Adoption Subsidy Issues," (Denver 2004).
American Academy of Adoption Attorneys Mid -Year Conference, "Adoption Subsidy Issues," (New Orleans 2004).
American Academy of Adoption Attorneys National Conference, Adoption in the Cradle of Liberty, "Adoption Subsidy Issues," (Philadelphia 2004).
Colorado Administrative Law Judges Conference, "Law and Practice of Adoption Subsidies," (Denver 2003).
Adoption Exchange, Adoption Education for Legal Professionals, "Stepparent, Custodial and Kinship Adoptions," (Denver 2003).
Colorado Office of the Child's Representative, Through the Child's Eyes, "Adoption Subsidies…Advocating for Children with Special Needs," (La Junta and Steamboat Springs 2003).
Oklahoma Lawyers for Children, "The Law and Practice of Adoption Subsidies," (Oklahoma City 2002).
National Association of Counsel for Children Annual Conference, "The Law and Practice of Adoption Subsidies," (New Orleans 2003).
Nevada Court Appointed Special Advocate Statewide Conference, "Adoption Subsides for Older Children," (Las Vegas 2003).
Timothy Eirich
Mr. Eirich frequently lectures throughout Colorado on child welfare and adoption related issues. A list of his trainings is below. Mr. Eirich has also served on the faculty of the National Institute of Trial Advocacy (NITA) since 2009.
Tim Eirich will serve as a facilitator and member of the planning committee for the Colorado Dependency & Neglect Judicial Institute in Boulder, Colorado. Tim will present to the judges and magistrates on "Intervention By Relatives and Foster Parents." (July 2015)
Tim Eirich and Katie Glynn presented with the Honorable Brett Woods of the Denver Juvenile Court at the Domestic Relations Best Practice Court Institute for Judges, Magistrates, Facilitators and Sherlocks in Denver on "Special Immigrant Juvenile Status." (June 2015)
Tim Eirich shared his insights with the Colorado State Foster Parent Association in Breckenridge, Colorado, on "Rights of Foster Parents and Relatives in Dependency & Neglect Cases" as well as "Adoption Subsidies for Special Needs Children." (June 2015)
Tim Eirich and Seth Grob presented to judges, magistrates and clerks of the 1st Judicial District on Contested Adoption Cases and Best Practices in Relinquishment Cases in Golden, Colorado. (June 2015)
"Intervention of Foster Parents and Relatives in Dependency and Neglect Cases", Denver, CO (December 2014), Juvenile Law Section of the Colorado Bar Association
"The Intersection of Family Law and Immigration Law" (with Seth A. Grob, Esq.), Denver, CO (September 2014), Colorado Bar Association, Immigration Law 2014
"Adoption Subsidies for Children with Special Needs," Denver, CO (May 2014), Colorado State Foster Parent Association, Annual Education Conference
"Legal Rights of Foster Parents and Caregivers in Colorado Dependency and Neglect Cases," Denver, CO (May 2014), Colorado State Foster Parent Association, Annual Education Conference
"Role of a GAL in the Child Welfare System," (with Betsy Fordyce, Esq.) Sheridan, CO (November 2013), Colorado State Foster Parent Association
"Case Law Update: Rights of Foster Parents in Dependency and Neglect Cases," Sheridan, CO (October 2013), Colorado State Foster Parent Association
"Impact of Adoption and ARTS on Domestic Relations," (with Laura Koupal, Esq.) Vail, CO (August 2013), Family Law Institute
"Role of Foster Parents in Dependency and Neglect Cases," Denver, CO (August 2013), Denver Juvenile Court Noon CLE
"Adoption: Considerations for GALs," (with Seth Grob, Esq.) Silverthorne, CO (July 2013), Office of Child's Representative Summer Conference
Family Law Institute, "Impact of Adoption on Domestic Relations," (Vail 2013)
Three day Basic Trial Skills course with the National Institute of Trial Advocacy, (Springfield, IL, June 2013).
Colorado State Foster Parent Association State Conference, "Foster Parent Rights in Colorado," (Breckenridge 2012).
Colorado State Foster Parent Association State Conference, "Adoption Subsidies for Children with Special Needs," (Breckenridge 2012).
Colorado County Attorney Association, "The Right to Intervene in Dependency and Neglect Cases," (Colorado Springs 2012).
The Children's Hospital – Neonatal Noon Conference, "The Medically Fragile Newborn: The Law and the Role of the Medical Professional," (Denver 2010).
Colorado State Foster Parent Association Bootcamp, "Role of the Guardian ad litem," (Centennial 2010).
University of Colorado School of Law presents The Legal System's Response to Domestic Violence, "The Role of Court Appointed Special Advocates," (Boulder 2009).
Colorado State Foster Parent Association State Conference, "Role of the Guardian ad litem," (Breckenridge 2009).
Colorado State Foster Parent Association State Conference, "Foster Parent's Rights in D&N Cases," (Glenwood Springs 2008).
Kathleen Glynn
Katie Glynn presented with others for an Immigrant Legal Resource Center webinar on "Strategies for Suppressing Evidence and Terminating Removal Proceedings for Child Clients." (May 2015)
Katie Glynn co-presented with Abbie Johnson of the Rocky Mountain Immigrant Advocacy Network (RMIAN), to child welfare professionals and judges at the Colorado Convening on Children, Youth, and Families in Beaver Creek. Their presentation focused on "The Intersection of Colorado State Child Welfare Law and Federal Immigration Law." (April 2015)
Faculty, Colorado Bar Association CLE: Immigration Law, "Representing the Most Vulnerable: Unaccompanied Children before the Immigration Court" (September 2014)
Faculty, Colorado Chapter of the American Immigration Lawyers Association (AILA) CLE: Asylum Law, "Children & Asylum" (November 2013)
Faculty, Kempe Center, 8th Annual National Differential Response Conference: Differential Response: A Catalyst for Change, "Working with Immigrant Children and Families in Differential Response" (October 2013)
Faculty, Colorado Juvenile Defender Coalition: Colorado Juvenile Defender Annual Conference, "Working with Non-Citizen Youth in Delinquency Proceedings: Pathways & Obstacles to Obtaining Immigration Status" (October 2013)
Faculty, Office of the Child's Representative: Webinar, "Immigration Issues for Court-Involved Youth and Families" (October 2013)
Faculty, Office of the Colorado State Public Defender CLE: "Working with Non-Citizen Youth in Delinquency Proceedings:
Pathways & Obstacles to Obtaining Immigration Status" (September 2013)
Faculty, Colorado County Attorneys Association: Annual Conference, "Working with Non-Citizen Youth and Families:
Pathways & Obstacles to Obtaining Immigration Status" (June 2013)
Faculty, Colorado Bar Association CLE: Immigration Law, "Representing Children in Immigration Proceedings," (May 2013)
Presenter, Adams County Human Services Department: 1st Annual Child Welfare Conference, Strengthening Families and Communities, "Working with Immigrant Children & Families: Obstacles & Opportunities on the Path to Lawful Status in the United States" (May 2013)
Faculty, Colorado Bar Association CLE: Juvenile Law, "Special Immigrant Juvenile Status (SIJS):A Pathway to Citizenship for Court Involved Youth" (April 2013)
Contact Grob & Eirich, LLC, to speak with a thoughtful, thorough adoption attorney with a passion for helping to build families.
Contested Adoption
Adoption Dissolution
Adoption Subsidy Negotiation
Adoption Agency Representation
Funding An Adoption
Assisted Reproductive Technology Resources
Immigration Options for Families
Citizenship Claims
Family-Based Petitions And Adjustment Of Status
Removal Defense
Guardianship and Custody
12596 W. Bayaud Avenue Suite 390
| Lakewood, CO 80228 | P: 303-816-8147
Additional offices in Boulder, Cherry Creek, Denver, Denver Tech Center and Evergreen.
© 2021 Grob & Eirich, LLC. All Rights Reserved.

redpajama_set_name: RedPajamaCommonCrawl


Black Foxxes have enjoyed a near explosive start to their career.
The band's first music video went online a mere three weeks ago, and the response has been truly incredible.
Gaining plays on Radio 1 – Huw Stephens and Annie Mac are both fans – it seems that the newcomers have truly hit a chord.
Debut album 'I'm Not Well' arrives on August 19th, following a summer of live commitments that will take Black Foxxes across the country and beyond.
Clash is able to premiere a new video from Black Foxxes, offering a short preview of their debut album.
Crisp riffs, taut songwriting and killer choruses – what's not to love? Tune in below.
'I'm Not Well' arrives on August 19th.


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Stayzilla
Yogendra Vasupal
Nasscom
iSpirt
IAMAI
Industry associations call for Stayzilla CEO's release & guidelines for startup closures
Industry associations such as FICCI, NASSCOM, iSPIRT, CII, IAMAI and TIE have asked for Vasupal's release apart from a right legal framework for business closures.Mugdha Variyar | ET Bureau | Updated: March 23, 2017, 05:47 IST
With Stayzilla founder Yogendra Vasupal failing to get a bail after over a week in jail in Chennai, several industry associations have come together to appeal for his release and to highlight the negative repercussions of this case.
Several entrepreneurs and investors who have formed a group post Vasupal's arrest have also been warning of the impact on the startup culture in the country.
Industry associations such as FICCI, NASSCOM, iSPIRT, CII, IAMAI and TIE issued a press note on Wednesday, and aside from asking for Vasupal's release, they also called for a right legal framework for business closures.
"Yogendra continues to be in jail in Chennai for over a week with no sight of his bail applications coming up for hearing. This is very unfortunate and we strongly urge the relevant authorities in the state of Tamil Nadu to intervene on the issue and support an early hearing of the bail plea," leaders from the associations said in a joint statement.
The members who signed the letter include Kris Gopalakrishnan, chairman of CII Startup Council, A Didar Singh, secretary general at FICCI, Ravi Gururaj, executive council member at NASSCOM, Sharad Sharma, cofounder of iSPIRT, Shubho Ray, president of IAMAI, Naganand Doraswamy, governing council member of TiE Bangalore, and R. Narayanan, president of TiE Chennai
Vasupal, founder of Stayzilla that had announced the halt of its operations last month, was taken into custody by the Chennai police following charges of defraud by an advertising firm.
Earlier this week, a group of over 80 entrepreneurs and investors, including the likes of Paytm's Vijay Shekhar Sharma and Ola's Bhavish Aggarwal, had written to the home minister asking for his intervention in the case.
Many in the startup community who have rallied behind the Stayzilla founder have raised questions about what they say should have been a civil dispute that was turned into a criminal one.
"Start-ups have become the growth engine of innovation for India and the Prime Minister's vision for Startup India has given a tremendous boost to the youth of the country and their dream to be an entrepreneur. However, as startups grow and scale, they will pivot, change priorities or close down.
Hence it is important that the right legal framework is provided to address any business closures. Any actions that go beyond the provisions of the law to create undue fear and harassment to the founders and their families as have been reported in this case, would raise the insecurity for startup founders, and make the cost of failure unacceptable to the vision of Startup India. This will set a very undesirable precedent," the industry associations said in their statement.
Tags : Startups, Stayzilla, Yogendra Vasupal, Nasscom, iSpirt, IAMAI, Government, FICCI, Entrepreneurship


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A$AP Ferg Drops Two New Freestyles
April 28, 2016 April 28, 2016 Tobias Handke
Currently out and about promoting his star studded sophomore album Always Strive And Prosper (read our review here), A$AP Ferg stopped by DJ Self's radio show at Power 106 and dropped a couple of freestyles live on air.
The Harlem rapper went in over The Lox's Money, Power & Respect and Diddy's Victory, referencing everything from Madonna, his riches, and Rihanna, to his position in the rap game. Both freestyles are great, and although Self's constant yelling and sound effects are as annoying as hell, once you get past them you realise how great a wordsmith Ferg is. Watching him getting animated and hyped as he's rapping is great, as you can feel the passion and love he has for his craft.
If there's one criticism, it's that Ferg already used portions of the freestyles when visiting The Cruz Show last month. During the show he was asked to freestyle over Wu-Tang Clan's Triumph, using much of the same rhymes as he did for the Money, Power & Respect freestyle. While this is disappointing and makes you wonder if any freestyles are actually off the cusp or are all written down beforehand, it doesn't take away from Ferg's writing, while his delivery is excellent.
Image: Billboard
Videos A$AP Ferg. permalink.
Listen: AlunaGeorge & ZHU Collaborate On "My Blood"
Watch: Flatbush Zombies New Joint Video For "Smoke Break" & "Fly Away"


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North Yorkshire, Moors and Coast
York man dead after car crash near Thirsk
The A168 near Thirsk. Picture: Google.
Ben Barnett
Published: 11:39 Sunday 16 December 2018
A man from York has died following a collision on the A168 near Thirsk.
It happened within a section of roadworks on the northbound side of the road, south of the junction with the A170 at around 6.45pm on Friday.
The driver of a black Vauxhall Insignia, aged in his mid-40s, was pronounced dead at the scene.
North Yorkshire Police said the man's family had been informed and were being supported by officers.
Diversions were put in place by the Highways Agency after the collision as emergency services responded to the crash. The road re-opened at around 12.30am yesterday.
Officers are appealing for people to come forward who witnessed the collision or recall seeing the black Vauxhall Insignia prior to the crash.
People with vehicle dash-cams who were travelling in the area at around the same time of the collision are also urged to check their footage for possible sightings.
Anyone who can assist police is asked to call the force on 101, quoting reference number 12180232472.
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Sheffield United expected to break club transfer record 'a few more times'


Here, we describe the ex vivo expansion of hematopoietic stem cells from CD34+ cells derived from umbilical cord blood treated with a combination of a cytokine cocktail and VPA. This method leads to a significant degree of ex vivo expansion of primitive HSCs for either clinical or laboratory applications.
Papa, L., Djedaini, M., Hoffman, R. Ex Vivo Expansion of Hematopoietic Stem Cells from Human Umbilical Cord Blood-derived CD34+ Cells Using Valproic Acid. J. Vis. Exp. (146), e59532, doi:10.3791/59532 (2019).
Umbilical cord blood (UCB) units provide an alternative source of human hematopoietic stem cells (HSCs) for patients who require allogeneic bone marrow transplantation. While UCB has several unique advantages, the limited numbers of HSCs within each UCB unit limits their use in regenerative medicine and HSC transplantation in adults. Efficient expansion of functional human HSCs can be achieved by ex vivo culturing of CD34+ cells isolated from UCBs and treated with a deacetylase inhibitor, valproic acid (VPA). The protocol detailed here describes the culture conditions and methodology to rapidly isolate CD34+ cells and expand to a high degree a pool of primitive HSCs. The expanded HSCs are capable of establishing both short-term and long-term engraftment and are able to give rise to all types of differentiated hematopoietic cells. This method also holds potential for clinical application in autologous HSC gene therapy and provides an attractive approach to overcome the loss of functional HSCs associated with gene editing.
Ex vivo expansion of hematopoietic stem cells (HSCs) from umbilical cord blood (UCB) units holds great promise for HSC applications in regenerative medicine and transplantation therapy. Transplantation with UCB units has several unique advantages such as easy collection, high availability, minimal risk of infection, low risk of disease relapse, and low frequency of graft-versus host disease (GVHD). However, the major disadvantages of their use in clinical settings are the limited number of HSCs present within each UCB unit1. The insufficient number of HSCs results in delayed engraftment and hematopoietic recovery, risk of graft rejection, and aberrant immune reconstitution.
Currently, various methods and strategies have been developed to ex vivo expand the limited number of HSCs from UCBs. Combinations of different cytokine cocktails with small molecules or compounds in ex vivo cultures result in various degrees of expansion in HSC numbers2,3,4,5,6,7,8. Importantly, ex vivo culture conditions induce stress, leading to rapid cell proliferation, increased metabolic activity and loss of the primitive characteristics that define primary HSCs. Therefore, developing protocols that lead to expansion of a great number of functional HSCs with characteristics that closely resemble primary primitive HSCs are needed.
Serum-free cultures of CD34+ cells isolated from UCBs and treated with valproic acid (VPA) result in the expansion of large numbers of primitive HSCs4,9,10. The HSC expansion is not solely due to the proliferation of the existing HSCs derived from UCBs. Instead, this expansion is due to the acquisition of a primitive phenotype combined with a limited number of cell divisions and proliferation9. Within the initial 24–48 h of incubation with a combination of cytokines and VPA, CD34+ cells acquire a transcriptomic and phenotypic profile that characterize long-term HSCs. The significant increase in the percentage of HSCs is accompanied by a prompt increase in the number of HSCs (63 fold increase within 24 h of VPA treatment)9. Notably, the VPA-ex vivo expansion strategy expands HSCs with low metabolic activity, which further highlights their primitive characteristics.
The method described here provides conditions and treatments that lead to a significant degree of ex vivo expansion of primitive HSCs for either clinical or laboratory applications. This ex vivo expansion strategy uses a cytokine cocktail combined with VPA treatment. VPA is an FDA approved drug for treatment of bipolar disorders and other neurological diseases. The HSC expansion with VPA is prompt and occurs within 7 days, minimizing both the time of manipulation and the risk of contamination. Importantly, this protocol allows for the acquisition of long-term HSC phenotypic markers such as CD90 and CD49f within 24-48 h following treatment with VPA9. Expanded HSC grafts created with this protocol have the capacity to regenerate the hematopoietic system since they can differentiate into all hematopoietic cell lineages and establish long-term engraftment following transplantation into myeloablated NSG mice models4. Moreover, this protocol is highly reproducible and allows for efficient and rapid isolation of viable CD34+ cells from UCBs, which is critical for industrialization of this procedure.
The VPA ex vivo expansion protocol also has the potential to overcome the significant loss of HSCs which occurs during gene editing11. Gene editing requires exposure to cytokines, which are necessary for cycling cells and activation of DNA-repair mechanisms. Due to the prompt effects of VPA treatment, this method might be beneficial for the generation of a higher number of genetically modified cells within a period of time that is relevant for currently utilized gene modification protocols.
The HSC ex vivo expansion protocol follows the guidelines of the Research Ethics Committee at Mount Sinai School of Medicine.
Prepare separation buffer 24 h prior to isolation of CD34+ cells from UBCs by adding 2 mL of 0.5 M Ethylene Diamine Tetra-acetic Acid (EDTA) and 33 mL of 7.5% Bovine Serum Albumin (BSA) to 465 mL buffered saline (1x PBS). Mix gently and maintain the buffer overnight at 4 °C.
Warm media at room temperature on the day of purification.
Dispense the whole UCB unit into a 75 cm2 flask. Dilute and gently mix the UCB with an equal volume of PBS at room temperature.
Determine the number of tubes required for processing the whole UCB unit (one 50 mL conical tube can be used to process 35 mL of diluted UCB). Add 15 mL of density gradient media (see Table of Materials) to each tube and dispense the diluted UCB on the top of the density gradient media creating two layers.
NOTE: Dispense the diluted UCB very slowly while holding the tube at a 45° angle to prevent mixing of the density gradient media layer with the UCB.
Centrifuge at 400 x g for 30 min at a low acceleration and deceleration rate. After centrifugation, MNCs locate in the white layer (buffy coat) between the plasma and density gradient media layer.
Slowly aspirate 2/3 of the plasma without disturbing the buffy coat layer. Carefully transfer the buffy coat layer into a new 50 mL tube. Collect all MNCs from the same UCB unit into a 50 mL tube until reaching 25 mL. Use a new tube if the volume of collected MNCs exceeds 25 mL.
Add 25 mL of cold PBS into the tube containing 25 mL of MNCs and mix well. Centrifuge at 400 x g for 10 min at 4 °C.
NOTE: Cold PBS is important to prevent capping of antibodies on the cell surface and reduce non-specific labeling.
Carefully aspirate the supernatant and gently re-suspend the cell pellet in 50 mL of cold PBS.
Remove 20 μL of the cell suspension for counting. Count the cell number stained with acridine orange/propidium iodide staining by using an automated cell counter. Calculate the total number of MNCs.
NOTE: The cell count can be also performed by the trypan blue exclusion method using a hemocytometer.
Centrifuge at 400 x g for 15 min at 4 °C.
NOTE: If not required immediately, MNCs can be frozen at this stage.
Prepare the CD34+ antibody coupled with magnetic beads solution for isolation of CD34+ cells from MNCs. Mix 300 µL of separation buffer with 100 µL of human FcR human IgG (blocking reagent) and 100 µL of CD34 magnetic beads for isolation of CD34+ cells from each 108 MNCs. For isolation of a higher number of CD34+ cells from (>108) MNCs, scale up reagents accordingly.
Carefully aspirate the supernatant from the tube centrifuged at step 2.8. Resuspend the pellet in the CD34 magnetic beads solution prepared in step 3.1 (use 500 µL of solution per 108 cells).
Mix gently and incubate at 4 °C for 30 min.
NOTE: Prepare the cell separator device (see Table of Materials) during the incubation period. Replace the storage solution with the working buffer as indicated by the manufacturer's instructions and run a washing program followed by a rinsing program.
Add cold cell separator running buffer to the tube containing the cell mixture until the tube is fully filled. Centrifuge at 400 x g for 15 min at 4 °C.
Aspirate the supernatant and resuspend the cell pellet in cold cell separator running buffer (2 mL/108 cells). Transfer the 2 mL of cell suspension mixture into 15 mL tubes.
Load 3 sets of 15 mL tubes into the cell separator rack prechilled at 4 °C. One set of tubes contains MNCs, the second set of tubes will be used to collect the negative fraction and the third set of tubes will be used to collect purified CD34+ cells.
Load the rack into the cell separator device and run the posseld2 preset program.
NOTE: An alternative method that utilizes manual magnetic separator can be used to replace the cell separator device12 .
Centrifuge tubes with the positive fraction at 400 x g for 15 min at 4 °C. Aspirate the supernatant and resuspend purified CD34+ cells in 1 mL of serum-free media.
Count the cells stained with acridine orange/propidium iodide using an automated cell counter.
NOTE: If not required immediately, purified CD34+ cells can be frozen at this stage.
Prepare sufficient volume of media to plate the purified CD34+ cells at a density of 3.3 x 104 cells/mL. The culture media composition is serum Free culture medium (see Table of Materials) supplemented with 10 µL/mL pen/strep, 150 ng/mL stem cell factor (SCF), 100 ng/mL fms-like tyrosine kinase receptor 3 (FLT3 ligand), 100 ng/mL thrombopoietin (TPO), and 50 ng/mL interleukin 3 (IL-3).
Plate purified CD34+ cells in a 12-well plate (5 x 104 CD34+ cells/ 1.5 mL of media/ well) and culture at 37 °C in a 5% CO2 humidified incubator.
Assess the purity of isolated CD34+ cells and cell composition by FACS analysis as described below in step 6.3.
Add VPA at a final concentration of 1 mM into the cultures of cells treated for 16 h with cytokine cocktail.
Culture the cells for an additional 7 days at 37 °C in a 5% CO2 humidified incubator.
NOTE: Media remains unchanged during the duration of the ex vivo expansion.
Prepare antibody-staining solution to stain 2 x 104–6 x 104 cells and isotype solution to determine FACS gating strategy and determine the level of non-specific binding. Dilute antibodies (1/100 APC anti-CD34, 1/200 FITC anti-CD90) and the respective isotypes into 50 µL of separation buffer (composition of separation buffer is defined in step 1.1).
NOTE: Perform single staining of cells with each antibody for a FMO gating strategy. Total antibody compensation bead kit (see Table of Materials) can be used for compensation setup.
Homogenize the cell culture by pipetting multiple times. Count cells and pipet 5 x 104 cells into a 1.5 mL tube.
Add 1 mL of separation buffer and centrifuge at 400 x g for 15 min at room temperature.
Aspirate the supernatant and resuspend the pellet in 50 µL of the staining solution. Incubate cells for 30 min at room temperature.
Add 450 µL of separation buffer and centrifuge at 400 x g for 15 min at room temperature.
Aspirate the supernatant and resuspend the pellet in 100 µL of separation buffer. Keep cells on ice for at least 5 min until performing FACS analysis. Add 1 µL of 7-AAD to gate viable cells.
Load the stained cell sample on the FACS machine and acquire cells at the lowest flow rate.
For each fluorophore, analyze the isotype controls and single stained cells to set up gating for FITC (CD90), APC (CD34), and PerCP/Cy5.5 (7-AAD) staining.
Gate PerCP/Cy5.5 negative fraction and plot APC vs FITC to determine the percent of viable CD34+, CD90+, and CD34+CD90+ cells that defines the HSPC/HSC population in the culture.
Resuspend the expanded cells by pipetting multiple times. Count the cell number and transfer the whole culture into a 15 mL or 50 mL tube.
Prepare antibody-staining solution to stain 5 x 105–2 x 106 cells and isotype solution to stain 1 x 105 cells and determine FACS gating. Dilute 1/10 (APC anti-CD34), 1/20 (FITC-CD90) antibodies and the corresponding isotypes into 500 µL of separation buffer per each condition.
Prepare the single color compensation controls using a total antibody compensation bead kit according to manufacturer's instructions.
Transfer the supernatant from step 5.2 into a new tube and keep it at 37 °C. This culture media will be reused to culture the sorted cells.
Resuspend the cell pellet in cold separation buffer to obtain a concentration of 5 x 105 cells/ mL. Transfer 1 x 105 cells in 1.5 mL tubes for isotype staining and the remaining cells that will be sorted into other 1.5 mL tubes.
Centrifuge the tubes at 400 x g for 15 min at 4 °C.
Discard the supernatant and resuspend pellets of cells in isotype staining solution and the pellet of cells that will be sorted in the antibody staining solution.
Add 450 µL of separation buffer and centrifuge at 400 x g for 15 min at 4 °C.
Resuspend the pellets with 2 mL of cold separation buffer.
To prevent clogs, filter the samples through a 40 μm cell strainer and place on ice until FACS.
Set-up cell sorter for sorting with the correct parameters.
Run isotype samples to set voltage and gain for forward and side scatter and identify negative fluorescence populations.
Run single color controls to set compensation coefficients and apply compensated parameter to collection protocol.
Run a small aliquot of the (Ab) sample (~50,000 events) to establish the gating strategy.
Set up sort decisions to collect the CD34+ CD90- cell fraction.
Sort cells into collection tubes coated with 2 mL separation buffer.
After sorting, recount the cells and centrifuge them at 400 x g for 15 min at 4 °C.
Discard the supernatant and resuspend the pellet in the media recovered in step 5.5 to reach a final concentration of 3 x 104 cells/mL.
Plate CD34+ CD90- purified cells in 12-well plates with 1.5 mL medium/well (5 x 104 CD34+ CD90- cells/1.5 mL of media/well).
Assess the purity by FACS analysis using 50 µL of media containing cells.
Treat the cultures of CD34+CD90- cells with VPA at 1mM final concentration.
Incubate at 37 °C in a 5% CO2 humidified incubator.
The ex vivo protocol described here increases the number of primitive HSCs generated from CD34+ cells isolated from UCBs (Figure 1). Priming of CD34+ cells for 16 h with a cytokine cocktail, followed by treatment with VPA for an additional 7 days, leads to a great degree of HSC expansion. Remarkably, the pool of expanded cells is highly enriched for HSCs, which are phenotypically defined by CD34+CD90+ markers. The total number of nucleated cells (TNCs) in cultures treated with the cytokine cocktail alone is significantly higher than the numbers of cells observed in cultures treated with the cytokine cocktail and VPA (Figure 2A). Despite the higher number of TNCs, the number of HSCs in the cultures receiving the cytokine cocktail alone remains low during the entire expansion period compared to cultures treated with the cytokine cocktail and VPA (Figure 2B). The greatest number of HSCs is generated in cultures treated with a combination of the cytokine cocktail and VPA (Figure 2B). In particular, the greatest expansion in the numbers of HSCs is reached after 5–7 days following VPA treatment (Figure 2B). The increased number of HSCs correlates with a prompt increase in the percentage of HSCs, which is notable within 24 h of VPA treatment (Figure 2C). While the increase in the percentage of HSCs is high and maintained during the first 4 days of ex vivo culture, it declines progressively after 5–7 days of treatment with VPA (Figure 2C). However, this decrease is inversely correlated with an increase in the absolute number of HSCs.
Importantly, the rapid increase in the percentage of HSCs observed in VPA treated cultures is due to the acquisition of the CD90 phenotype. CD34+ cells expressing the CD90 phenotypic marker reaches almost 40%–45% of cells expanded in ex vivo cultures treated with the cytokine cocktail and VPA for 4 days (Figure 2C,D). The acquisition of the CD90 phenotype is further confirmed by ex vivo expansion of the highly purified CD34+ cells that lack expression of the CD90 marker. Within 24 h of VPA treatment, almost 75% of the ex vivo expanded cells in cultures initiated with sorted CD34+CD90- cells express CD90 as opposed to 0% of the cells in cultures containing the cytokine cocktail alone (Figure 2E). Importantly, the CD90 phenotype is highly retained during the first 4 days in ex vivo cultures treated with VPA (Figure 2E).
Figure 1: Schematic presentation of ex vivo expansion culture. Purified CD34+ cells from UCBs are primed for 16 h in ex vivo culture supplemented with a combination of the indicated cytokines. Cultures are treated with VPA (1mM) for an additional 7 days. The ex vivo expanded cells may then be transplanted into myoablated NSG mice. Please click here to view a larger version of this figure.
Figure 2: VPA treatment triggers the acquisition of CD90 resulting in the expansion of a great number of HSCs. (A) Absolute number of total viable nucleated cells expanded in ex vivo cultures9. Purified CD34+ cells from UCBs described in section 3 treated with cytokine cocktail alone or a combination of cytokine cocktail and VPA as described in section 4 were counted by acridine orange/propidium iodide staining (n = 16). Numbers denote days of treatment with VPA, whereas PC denotes the primary uncultured CD34+ cells isolated from UCBs. (B, C) Absolute number and percentage of CD34+CD90+ cells expanded throughout 7 days in ex vivo cultures treated with cytokine cocktail alone or a combination of cytokine cocktail and VPA (n = 21). Percentage of CD34+CD90+ cells is determined by flow cytometry analysis of cells stained as described in section 4. (D) Phenotypic analysis of sorted CD34+ cells from UCBs (step 5.3) expanded in ex vivo cultures treated as indicated for 4 days. Left panel indicates the gating strategy using isotype staining whereas the other panels indicate stained cells expanded in cultures containing cytokine cocktail alone or a combination of cytokine cocktail with VPA. Numbers denote the percentage of CD34-CD90+ cells, CD34+CD90- cells and CD34+CD90+ cells. (E) Percentage of CD34+CD90+ cells expanded in cultures initiated with sorted CD34+CD90- cells from UBCs and treated with cytokine cocktail alone or a combination of cytokine cocktail and VPA for the indicated days. Percentage is determined by flow cytometry analysis (n = 4). N: number of biological replicates. Error bars with SEM; ****p ≤ 0.0001 as determined by negative-binomial models for A and B, Beta models for D and 2-way ANOVA for panel E. Panels A–C and E have been modified from Papa et al.9 Please click here to view a larger version of this figure.
Herein, we present a protocol to rapidly expand to a significant degree the number of functional human HSCs from UCBs. The pilot and kinetic studies using this protocol indicate that the ex vivo expanded cells promptly acquire and retain the expression of several HSC phenotypic markers including CD90 as well as primitive HSC metabolic characteristics9.
This ex vivo expansion protocol is relatively simple and reliable. Purification of CD34+ cells with the cell separator device (see Table of Materials) is highly reproducible and rapid, allowing for fast recovery of isolated cells. This method results in a high yield of purified CD34+ cells (>90%) as opposed to manual immunomagnetic separation. Moreover, this ex vivo expansion protocol does not require special and complex devices or fed-batch culture systems that need continuous supplies of large volumes of fresh media supplemented with cytokines3,13. Accordingly, this method limits costs. Importantly, this protocol allows for the expansion of the HSC pool within a short time, which is a key limiting factor for contamination frequency.
Several points should be considered for achieving primitive HSCs from CD34+ cells using this protocol. CD34+ cells expanded for 4 days in cultures treated with the cytokine cocktail and VPA lead to the generation of a pool of primitive long-term HSCs. These HSCs are characterized not only by the high expression levels of CD34, CD90 and CD49f, but also by a very primitive metabolic profile, which predominantly relies on glycolysis9. During more prolonged incubation with VPA for 7 days, expanded cultures however are more heterogeneous and contain a greater number of both long-term and short-term as well as differentiated cells as opposed to cells expanded for 4 days. This outcome is mainly due to the exhaustion of VPA in culture and increased number of cell divisions9. Thus, this protocol can be used to achieve expansion of two different pools of cells: 4-day and 7-day expanded cells. The 7-day expansion product can be used for allogeneic HSC transplantation where more rapid as well as sustained multilineage engraftment is required. The 4-day expansion protocol might be best suited for genetic modification strategies that target long-term repopulation cells. It is also likely that this combination of VPA and cytokine cocktail can expand the pool of HSCs from CD34+ cells at the same degree or higher when using media other than the media (see Table of Materials) used in this protocol.
Current gene editing procedures are associated with a significant loss of HSCs, which limit their clinical application. The great number of primitive HSCs achieved within 2–4 days of VPA treatment in ex vivo cultures has the potential to overcome this loss in HSC numbers. Thus, this protocol might be beneficial for improving the existing gene editing protocols and enable correction of genetic mutations in HSC transplantation-based therapies for β-thalassemia and sickle cell disease. Moreover, it can be applied to maintain great numbers of HSCs during gene manipulations for studies focused on the elucidation of gene functions in HSCs and in hematopoiesis.
In conclusion, the method described here can be used for ex vivo expansion of both human and murine HSCs and can be tailored to specific clinical applications as well as to a wide spectrum of investigations in basic research.
Broxmeyer, H. E. Enhancing the efficacy of engraftment of cord blood for hematopoietic cell transplantation. Transfusion and Apheresis Science. 54, (3), 364-372 (2016).
Boitano, A. E., et al. Aryl hydrocarbon receptor antagonists promote the expansion of human hematopoietic stem cells. Science. 329, (5997), 1345-1348 (2010).
Fares, I., et al. Cord blood expansion. Pyrimidoindole derivatives are agonists of human hematopoietic stem cell self-renewal. Science. 345, (6203), 1509-1512 (2014).
Chaurasia, P., Gajzer, D. C., Schaniel, C., D'Souza, S., Hoffman, R. Epigenetic reprogramming induces the expansion of cord blood stem cells. Journal of Clinical Investigation. 124, (6), 2378-2395 (2014).
Wagner, J. E. Jr, et al. Phase I/II Trial of StemRegenin-1 Expanded Umbilical Cord Blood Hematopoietic Stem Cells Supports Testing as a Stand-Alone Graft. Cell Stem Cell. 18, (1), 144-155 (2016).
Peled, T., et al. Nicotinamide, a SIRT1 inhibitor, inhibits differentiation and facilitates expansion of hematopoietic progenitor cells with enhanced bone marrow homing and engraftment. Experimental Hematology. 40, (4), 342-355.e1 (2012).
Nikiforow, S., Ritz, J. Dramatic Expansion of HSCs: New Possibilities for HSC Transplants? Cell Stem Cell. 18, (1), 10-12 (2016).
Mehta, R. S., et al. Novel Techniques for Ex vivo Expansion of Cord Blood: Clinical Trials. Frontiers in Medicine. 2, 89 (2015).
Papa, L., et al. Ex vivo human HSC expansion requires coordination of cellular reprogramming with mitochondrial remodeling and p53 activation. Blood Advances. 2, (20), 2766-2779 (2018).
Papa, L., Djedaini, M., Hoffman, R. Mitochondrial Role in Stemness and Differentiation of Hematopoietic Stem Cells. Stem Cells International. In press (2019).
Genovese, P., et al. Targeted genome editing in human repopulating haematopoietic stem cells. Nature. 510, (7504), 235-240 (2014).
Perdomo, J., Yan, F., Leung, H. H. L., Chong, B. H. Megakaryocyte Differentiation and Platelet Formation from Human Cord Blood-derived CD34+ Cells. Journal of Visualized Experiments. (130), e56420 (2017).
Fares, I., et al. EPCR expression marks UM171-expanded CD34(+) cord blood stem cells. Blood. 129, (25), 3344-3351 (2017).


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text:
These Hollywood Celebrities Have Spend Their Time In Jail, List Is Surprising
Vijay Alagar December 9, 2017
With a lot of money comes in a lot of bad influence and ideas. Keeping moral no matter how high you reach in life is very essential and a few fail terribly. These Hollywood celebrities are no exception to that. Listed below are a few Hollywood Celebs who had to go to Jail!
1. Lil Kim
The famous rapper was sentenced to a year and a day in 2005 for lying to the federal jury about her friends' involvement in a shootout near a Manhattan radio station in 2001. She was also fined $50,000.
2. Martha Stewart
During a corporate fraud investigation in 2004, the famous Chef and TV personality served in jail for five months for the charges of conspiracy, obstruction of justice and giving false statements to the authority.
3. Lindsey Lohan
The actor/singer is no new to controversies. She pleads guilty to driving under the influence of cocaine and was sentenced to a day in jail and 10 days of community service and three years of probation.
4. OJ Simpson
OJ Simpson had been in the news for all the wrong reasons, the majority of which that landed him in jail was on the charges of kidnapping and robbing two sports memorabilia dealers at gunpoint in a hotel room at Las Vegas.
5. Robert Downey Jr.
The Iron Man actor spent some quality time in jail for violating probation in 1996 for drug and weapon conviction. After about a year of serving prison time, he was given bail.
6. Paris Hilton
The "It" girl spent three days of prison period when she failed to enrol in a rehab program ordered by the court.
7. Mike Tyson
The heavyweight champion was found guilty of raping an eighteen-year-old beauty pageant and two other charges in the year 1992. At first, he was sentenced a ten year prison time and was later reduced to six.
8. 50 Cent
Curtis Jackson has spent about six months in prison for the possession of crack cocaine. Heroin and a starter gun in 1994. He later took up his nickname "50 Cents" as a metaphor for his change.
9. Michelle Rodriguez
In 2007 the Fast and Furious star failed to complete her community service hours following a DUI and so was sentenced to serve 18 days in jail, which she did.
10. Paul McCartney
The famous Paul McCartney of Beatles was arrested for the possession of Marijuana in Japan for a period of nine days in the year 1980.
I guess money doesn't help you evade the law always! These celebrities and much more are an example to it. Abide by the law and stay trouble free!
Love reading celebrity stories? Well, you have come to just the right place. Read more about these supporting actors from the 90s whom we miss the most.
These Ekta Kapoor TV Shows Are Every 90s Kids' Guilty Pleasure
These Movies Have Sold The Maximum Number Of Tickets In Indian Cinema
the authorVijay Alagar
Young and Ambitious; Better at Videogames than at life. Creative writer at daytime, Batman at night.
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text:
import os
# ********************PyQt5相关模块导入********************
from PyQt5.QtCore import Qt
from PyQt5.QtCore import QTranslator
from PyQt5.QtGui import QImage
from PyQt5.QtGui import QPixmap
from PyQt5.QtWidgets import qApp
from PyQt5.QtWidgets import QApplication
from PyQt5.QtWidgets import QInputDialog
from PyQt5.QtWidgets import QSplashScreen
# ********************用户自定义相关模块导入********************
from LaserQt_MainWindow import LaserQtMainWindow
from LaserQt_SecondWindow import LaserQtSecondWindow
from LaserQt_ThirdWindow import LaserQtThirdWindow
from LaserQt_FourthWindow import LaserQtFourthWindow
from LaserQt_ImageWindow import LaserQtImageWindow
from LaserQt_Gui.LaserQt_Gui_Dialog import MessageDialog

'''
@author  : Zhou Jian
@email   : zhoujian@hust.edu.cn
@version : V1.0
@date    : 2017.02.22
'''

class OverLoadClassMethod(object):
    '''
        利用该类分别对LaserQt_MainWindow.py、LaserQt_SecondWindow.py、LaserQt_ThirdWindow.py、LaserQt_FourthWindow.py中的部分按钮动态得绑定回调函数
    '''
    def __init__(self):
        super(OverLoadClassMethod, self).__init__()

    # LaserQt_MainWindow.next_page方法
    def laser_qt_main_window_next_page(self):
        myLaserQtMainWindow.hide()  # 隐藏第一个窗口
        myLaserQtSecondWindow.show()  # 显示第二个窗口

    # LaserQt_MainWindow.prev_page方法
    def laser_qt_second_window_prev_page(self):
        myLaserQtSecondWindow.hide()
        myLaserQtMainWindow.show()
        myLaserQtMainWindow.directoryLineEdit.setText("")
        myLaserQtMainWindow.dataTable.clearContents()
        myLaserQtMainWindow.dataTable.setRowCount(0)

    # LaserQt_SecondWindow.next_page方法
    def laser_qt_second_window_next_page(self):
        myLaserQtSecondWindow.hide()
        myLaserQtThirdWindow.show()

    # LaserQt_SecondWindow.prev_page方法
    def laser_qt_third_window_prev_page(self):
        myLaserQtThirdWindow.hide()
        myLaserQtSecondWindow.show()

    # LaserQt_ThirdWindow.next_page方法
    def laser_qt_third_window_next_page(self):
        messageDialog = MessageDialog()
        reply = messageDialog.question(myLaserQtThirdWindow, "消息提示对话框", "您是否已完成点云数据处理?", messageDialog.Yes | messageDialog.No, messageDialog.Yes)
        # 在进入第四个窗口前,先提示用户是否完成点云数据处理,因为第四个窗口的绘图数据依赖于第三个窗口所操作处理的数据
        if reply == messageDialog.No:
            return

        myLaserQtThirdWindow.hide()
        myLaserQtFourthWindow.show()
        # 将第三个窗口操作处理所产生的中间数据赋给第三个窗口,减少重复计算
        myLaserQtFourthWindow.matrix1 = myLaserQtThirdWindow.matrix1
        myLaserQtFourthWindow.matrix2 = myLaserQtThirdWindow.matrix2
        # 初始化第四个窗口的画布
        messageDialog = MessageDialog()
        reply = messageDialog.information(myLaserQtThirdWindow, "消息提示对话框", "可视化初始化!", messageDialog.Yes, messageDialog.Yes)
        if reply == messageDialog.Yes:
            myLaserQtFourthWindow.init_the_canvas()

    # LaserQt_FourthWindow.prev_page方法
    def laser_qt_fourth_window_prev_page(self):
        myLaserQtFourthWindow.hide()
        myLaserQtSecondWindow.show()

    # LaserQt_FourthWindow.enlarge_the_plot方法
    def laser_qt_fourth_window_enlarge_the_plot(self):
        global myLaserQtImageWindow
        # 提示用户输入所要放大显示的图像索引号
        num, isOk = QInputDialog.getInt(myLaserQtFourthWindow, "复杂曲率板加工系统", "输入待放大误差曲线图像序号", 1, 1, 6, 1)
        if isOk:
            if num == 1:
                myLaserQtImageWindow = LaserQtImageWindow("horizontal_direction_1_3_error_curve.png")
                myLaserQtImageWindow.show()
            elif num == 2:
                myLaserQtImageWindow = LaserQtImageWindow("horizontal_direction_1_2_error_curve.png")
                myLaserQtImageWindow.show()
            elif num == 3:
                myLaserQtImageWindow = LaserQtImageWindow("horizontal_direction_2_3_error_curve.png")
                myLaserQtImageWindow.show()
            elif num == 4:
                myLaserQtImageWindow = LaserQtImageWindow("vertical_direction_1_3_error_curve.png")
                myLaserQtImageWindow.show()
            elif num == 5:
                myLaserQtImageWindow = LaserQtImageWindow("vertical_direction_2_3_error_curve.png")
                myLaserQtImageWindow.show()
            # 用户自定义显示的图,必须先绘图,才能放大显示
            elif num == 6:
                if os.path.exists("LaserQt_Temp/between_two_arbitrary_point_error_curve.png"):
                    myLaserQtImageWindow = LaserQtImageWindow("between_two_arbitrary_point_error_curve.png")
                    myLaserQtImageWindow.show()
                else:
                    messageDialog = MessageDialog()
                    messageDialog.warning(myLaserQtFourthWindow, "消息提示对话框", "请先绘图!", messageDialog.Yes, messageDialog.Yes)

if __name__ == '__main__':
    import warnings
    warnings.filterwarnings('ignore')

    import sys
    app = QApplication(sys.argv)
    app.setApplicationDisplayName("v1.0")
    app.setApplicationName("LaserQt")
    app.setApplicationVersion("version 1.0")

    splash = QSplashScreen(QPixmap.fromImage(QImage("LaserQt_Ui/logo_128px.png")))
    splash.setDisabled(True)
    splash.show()

    # 读取样式表
    file = open("LaserQt_Gui/LaserQt_Gui_Style.qss", 'r')
    styleSheet = file.read()
    file.close()
    # 设置全局样式
    qApp.setStyleSheet(styleSheet)

    # 汉化处理
    tran = QTranslator()
    tran.load("qt_zh_CN.qm", "LaserQt_Font/")
    qApp.installTranslator(tran)

    myLaserQtMainWindow = LaserQtMainWindow()
    myLaserQtSecondWindow = LaserQtSecondWindow()
    myLaserQtThirdWindow = LaserQtThirdWindow()
    myLaserQtFourthWindow  = LaserQtFourthWindow()
    myLaserQtImageWindow = None

    # 对实例方法进行动态修改
    overLoad = OverLoadClassMethod()

    myLaserQtMainWindow.next_page = overLoad.laser_qt_main_window_next_page
    # 按钮绑定回调函数
    myLaserQtMainWindow.nextButton.clicked.connect(myLaserQtMainWindow.next_page)

    myLaserQtSecondWindow.prev_page = overLoad.laser_qt_second_window_prev_page
    myLaserQtSecondWindow.prevButton.clicked.connect(myLaserQtSecondWindow.prev_page)
    myLaserQtSecondWindow.next_page = overLoad.laser_qt_second_window_next_page
    myLaserQtSecondWindow.nextButton.clicked.connect(myLaserQtSecondWindow.next_page)

    myLaserQtThirdWindow.prev_page = overLoad.laser_qt_third_window_prev_page
    myLaserQtThirdWindow.prevButton.clicked.connect(myLaserQtThirdWindow.prev_page)
    myLaserQtThirdWindow.next_page = overLoad.laser_qt_third_window_next_page
    myLaserQtThirdWindow.nextButton.clicked.connect(myLaserQtThirdWindow.next_page)

    myLaserQtFourthWindow.prev_page = overLoad.laser_qt_fourth_window_prev_page
    myLaserQtFourthWindow.prevButton.clicked.connect(myLaserQtFourthWindow.prev_page)
    myLaserQtFourthWindow.enlarge_the_plot = overLoad.laser_qt_fourth_window_enlarge_the_plot
    myLaserQtFourthWindow.enlargeButton.clicked.connect(myLaserQtFourthWindow.enlarge_the_plot)

    # 显示第一个主窗口
    myLaserQtMainWindow.show()
    splash.finish(myLaserQtMainWindow)

    sys.exit(app.exec_())

redpajama_set_name: RedPajamaGithub


text:
Nancy Stewart will preform her concert series in various libraries throughout King County. Families can visit the KCLS website to find upcoming concerts. Courtesy of Nancy Stewart.
Nancy Stewart's 'A Season for Singing' concert coming to a library near you
King County Library System prepares for its annual Season of Stories and Songs.
by Stephanie Quiroz
Tuesday, November 27, 2018 1:30am
The King County Library System (KCLS) is hosting its annual Season of Stories and Songs. During this time of the year, KCLS hosts various holiday workshops around King County libraries that include stories, art and cooking workshops, music, puppet shows and more.
For musician Nancy Stewart, this is her favorite time of the year. Stewart has been a part of the KCLS's Season of Stories and Songs since 1996. Her winter concert series is called "A Season for Singing." Stewart released her recording of winter holiday songs for children in 1996. Since then, Stewart has preformed and shared the different holiday traditions of children around the world in King County libraries.
"For children, it's one of the most exciting times of the year. In every culture, there is something going on during this time of the year," said Stewart. "King County is very diverse. Every neighborhood library is very different."
Stewart shared how before she started her career, she noticed the link between singing and literacy. Every time parents sing with their children, they're getting them ready to read. With her own children, Stewart noticed the way her young children respond to music and how their language development increased, as well as their interest in literacy.
Her focus is to share the link between music and literacy. Stewart said a library setting is the perfect environment to do so. For her winter concert series, Stewart focuses on sharing the different holiday traditions of children around the world.
"[I was] trying to figure out a way to educate families and children about traditions…[I] was trying to do it in a respectful way and in a way that honors them," said Stewart. "It's tricky but that's where music is so effective. It really can cross all those barriers. Music is the thing that connects everyone."
Stewart will preform in various libraries throughout King County. She started off the holiday season with a concert in Skykomish on Nov. 16. Families can visit the KCLS website to find Stewart's upcoming concerts.
During her holiday concert series, families can expect to hear interesting snippets of each holiday tradition. Stewart tries to find a way to invite children to participate in the songs. Children can sing along to their favorite Christmas songs, and songs for Hanukkah, Kwanza and Chinese New Year. Stewart will play songs from her recording, "A Season for Singing" that include songs like "Frosty the Snowman," "My Dreydl," "Break The Piñata," "Oh Tannebaum" and more.
Stewart shared she learns something new every year. She likes adding different songs and using visual props. She created a blue backdrop decorated with Christmas trees decorated with animals and lights. Stewart believes lights and animals are "the somethings" that unite the holiday traditions all over the world.
"We live in a world, especially now where we need to know about each other," said Stewart. "The more we learn about each other, the more we realize we're really all the same and that we have so much in common."
Stewart shared she writes from a third person's view point. Earlier on, families would approach her to ask if she could write a song about their tradition. Stewart receives various emails and phone calls from mothers who are so grateful to have a song for their children. It means a lot to families to have their holiday traditions shared with everyone.
"It's the only time of the year that we're all celebrating something. Every celebration incorporates music," said Stewart.
Some of the traditions Stewart sings about include one from Norway. She shared how she brings a giant toothbrush to libraries so children can scrub from head to toe. Her song, "Rub A Dub Dub," sings "In Norway, so far away/All children gettin' ready for the holiday/Wash fingers and toes, elbows and nose/So when the holiday comes they're ready to go." Another fun tradition she sings about is one from England. Stewart shared how bees come out of their hives and sing to the children on Christmas Eve in England. While she sings, "Buzz buzz buzz, hear the buzzing of the bees/Singing little lullabies on Christmas Eve," participating children wear bumble bee head bands and buzz, "Tinkle Little Star."
Stewart is passionate about the library system and believes in the importance of singing. She said there is a connection between singing and literacy.
To learn more about Stewart's library programs and music workshops visit, nancymusic.com or email her at nancy@nancymusic.com.
KCLS will be hosting "A Season for Singing" concerts in 10 libraries throughout King County, starting Nov. 16 through the new year.
KCLS hosts various workshops throughout the holiday season. To find upcoming workshops, visit their event calendar at kcls.org.
King County Metro buses paused all bus services to honor memory of transit operator Mark McLaughlin
Award-winning play 'The Good Adoptee' coming to Mercer Island
The autobiographical drama was penned by acclaimed playwright Suzanne Bachner.
Scouts to recycle trees on Jan. 4
The 35th year of recycling trees for Sammamish and Issaquah Highlands residents.
HopeFest founder Tyler Zangaglia: "We have a responsibility to give back"
Zangaglia's experiences propelled him to launch his nonprofit, HopeFest.
New Year's Day will serve frigid dip
First ever Polar Plunge at Lake Sammamish State Park.
'It's one of my favorite places to be': School of Rock Issaquah gears up for January shows
In January, students will be paying homage to the Beatles, Black Sabbath, Chris Cornell and others.
Three simple rules for the holiday
A monthly column about mindfulness.
Issaquah to host its 10th Annual Turkey Trot 5K
The race begins at 9 a.m. on Nov. 28.
Turkeys for all on Nov. 23
Local church plans to give out 1,500 free turkeys.
One Million Trees Celebration with Green Issaquah
Volunteer events, new forest stewardship program.
Annual turkey giveaway on Nov. 23
Last year's event dished out about 1,500 turkeys.
Day of the Dead at Issaquah Highlands
Blakely Hall was full of bright colors, paper flowers, painted faces, sugar… Continue reading


Sandy Acres Farm is a third generation farm that has been in operation since 1958. Our high quality beef and pork are raised using all natural feed produced on the farm or bought locally.
In 1992, Sandy Acres Farm was recognized by the Thunder Bay Soil and Crop Association as a Soil and Water Conservation Farm because of our efforts on farm waste management. We also maintain 18 acres of untouched marsh and wetlands. This property includes 2 ponds and an abundance of waterfowl, plants and wildlife. Thank you for visiting our site and learning about us and where your food comes from. We look forward to hearing from you in the near future.


text:
August 1, 2014 by Melissa Fox
A Magical Evening Aboard the USS Iowa
I had the pleasure this past weekend of attending the first Western Region Sea Scouts Bridge of Honor, held aboard the battleship USS Iowa in the Port of Los Angeles.
A Bridge of Honor recognizes advancement and other achievements earned by Sea Scouts – the co-ed, nautical program of the Boy Scouts of America.
At this Bridge of Honor, several Sea Scouts were recognized for achieving the Quartermaster Award (the highest award in Sea Scouts) and many more were recognized for achieving the ranks of Apprentice, Ordinary and Able.
I was in attendance because my son, Max, received his rank of Ordinary, and because my husband serves as Commodore for our area of the Western Region Sea Scouts. I also serve as a Sea Scout adult volunteer for Sea Scout Ship 90 at the Newport Sea Base.
The early evening ceremony aboard the USS Iowa was breathe-taking. There could be no more majestic location than the deck of one of America's mightiest battleships, which earned nine battle stars for World War Two service and two battle stars for Korean War service, and which carried President Franklin D. Roosevelt, Admiral D. William Leahy, General George C. Marshall, Admiral Ernest King, and General Henry "Hap" Arnold to the Tehran Conference in the midst of World War II.
Along with more than 100 Sea Scouts from across the West, National Sea Scout Executive Keith Christopher was present, as was the Sea Scouts National Commodore (and retired Coast Guard Vice Admiral) Charles D. Wurster. I was tremendously proud when Admiral Wurster and Western Region Commodore Josh Gilliland congratulated Max.
I was also tremendously impressed by the enthusiasm, focus, patriotism, and camaraderie of these young Sea Scouts dressed in their best dress whites and dress blues — and by their love of the sea and adventure.
It was magical to watch them dancing in the twilight on the deck of the Iowa to the sounds of a live swing band.
Truly an amazing evening – with the USS Iowa to remind us of the greatness of our past and these young men and women of the Sea Scouts ready, eager and able to secure the greatness of our future.
This entry was posted in Uncategorized and tagged Admiral Charles D. Wurster, Admiral D. William Leahy, Admiral Ernest King, Battleship USS Iowa, Boy Scouts, Boy Scouts of America, Bridge of Honor, Coast Guard Vice Admiral Charles D. Wurster, Franklin D. Roosevelt, General George C. Marshall, General Henry "Hap" Arnold, Irvine Commissioner Melissa Fox, Josh Gilliland, Keith Christopher, Korean War, landship, Melissa Fox, melissafoxblog, melissafoxblog.com, melissajoifox, National Sea Scout Executive Keith Christopher, Newport Sea Base, Pacific Battleship Center, Port of Los Angeles, President Franklin D. Roosevelt, Quartermaster Award, Sea Scout landship, Sea Scouts, Sea Scouts Bridge of Honor, Sea Scouts National Commodore Charles D. Wurster, Tehran Conference, United States Coast Guard, United States Navy, US Coast Guard, US Navy, USS Iowa, Western Region Commodore Josh Gilliland, Western Region Sea Scouts, World War Two. Bookmark the permalink.
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Our culture inculcates several behaviors that harm creative, entrepreneurial people: we're taught to avoid conflict and real, open honesty with one another, and we're taught to fear failure. This may work for creating a polite, civil society, but it doesn't serve the individual who is trying to make a difference. But there's hope! Improv theater actors have devised many techniques for overcoming these tendencies which enables them to create inventive stories extemporaneously. I share ways to combat creative inhibitions gleaned from six years of teaching and performing improv theater.
Undoing Your Social Training and Creative Inhibitions in Five ... My talk was called "Undoing Your Social Training and Creative Inhibitions in Five Minutes".
... an online pinboard to collect and share what inspires you. No data available. Blogs (0) ... Social networks (14) Loading... No data available.
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Beautiful Bungalow in Elviria. The very well maintained property consist of 2 bedrooms, 2 bathrooms, lounge with dining area and kitchen in the main house and a third bedroom in the guest apartment with its own entrance and bathroom ensuite.
A spacious lounge with dining area is leading out of the big and partly covered terrace. Lovely views over the landscaped garden and the heated pool.
There is a garage for one car and a fully equipped kitchen.
The Bungalow is located in the lower part of popular Elviria and within walking distance to shops, restaurant and the beach.
A lot of potential for reform, another floor can be added and it can be extended by 60m2.


text:
Q: NancyFx and unit testing Response.AsFile(...) fails with NotFound This is my (streamlined) Module
public class MyModule : NancyModule
{
    public MyModule() : base("/path")
    {
        Get["/{filename}"] = x =>
        {
            var fileName = (string)x.filename;
            var path = Path.Combine("files", fileName);
            if (!File.Exists(path)) throw new FileNotFoundException();

            return Response.AsFile(path);
        }
    }
}

This code works and sucessfully delivers the correct file to the browser.
Now I have written a test:
    [TestMethod, DeploymentItem("file.txt", "files")]
    public void CanDownloadFile()
    {

        var path = @"files\file.txt";
        Assert.IsTrue(File.Exists(path));

        var bootstrapper = new MyBootstrapper();
        var browser = new Browser(bootstrapper); // from Nancy.Testing

        var result = browser.Get("/path/file.txt", with =>
        {
            with.HttpRequest();
        });

        var body = result.Body.AsString();
        Assert.AreEqual(HttpStatusCode.OK, result.StatusCode);
    }

With this code my test fails because the StatusCode is NotFound instead of OK
I even tried to replace Response.AsFile() with
var info = new FileInfo(path);
return new GenericFileResponse(info.FullName);

and triple checked the file really exists physically on disk wile executing my test.
What's the problem?

A: Is the file "file.txt" included in de Solution? Include this.
In the file properties in Visual Studio change the property Copy to Output Directory to Copy if newer.
But you don't need to create a Module to do this. Yo can manage static content in the Nancy bootstrapper. Here is the documentation https://github.com/NancyFx/Nancy/wiki/Managing-static-content
Doing this, you don´t need to test that.

redpajama_set_name: RedPajamaStackExchange


text:
\section{Constraining the $\rho$ wavefunction}

\vspace{2\baselineskip}

\centerline{\bf Abstract}
\vspace{0.7\baselineskip}
\parbox{0.9\textwidth}{Diffractive $\rho$ meson production has been identified
as one of the important
processes where saturation can be probed at a future Electron-Ion Collider
(EIC). A source of uncertainty in making predictions for this process lies
within the assumed form of the meson light-cone wavefunction. We report
here the results of reference~\cite{Forshaw:2010py} where a Regge-inspired
dipole
model was used to extract this wavefunction as well as the corresponding leading
twist-$2$ Distribution Amplitude from the current accurate HERA data. In
addition, we shall check the robustness of the conclusions in
reference~\cite{Forshaw:2010py} by using alternative Colour Glass Condensate
dipole
models. }

\section{Introduction}

In the dipole model~\cite{Nikolaev:1990ja,Mueller:1994jq}, the
imaginary part of the amplitude  for diffractive $\rho$ production is written
as~\cite{Marquet:2007qa}
\begin{equation}
\Im \mbox{m} \mathcal{A}_{\lambda} (s,t;Q^2) =
\sum_{h,\bar{h}} \int \mathrm{d}^2  \mathbf{r} \mathrm{d} z
\Psi^{\gamma^*,\lambda}_{h,\bar{h}} (r,z;Q^2)
\Psi^{\rho,\lambda}_{h,\bar{h}}(r,z)^{*}
e^{-iz\mathbf{r}.\mathbf{\Delta}} \mathcal{N}(x,\mathbf{r},\mathbf{\Delta})
\label{non-forward-amplitude}
\end{equation}
where $t=-|\mathbf{\Delta}|^2$. In a standard
notation~\cite{Forshaw:2010py,Marquet:2007qa,Forshaw:2003ki},
$\Psi^{\gamma^*,\lambda}_{h,\bar{h}}$ and $\Psi^{\rho,\lambda}_{h,\bar{h}}$ are
the light-cone wavefunctions of the photon and the $\rho$ meson respectively
while $\mathcal{N}(x,\mathbf{r},\mathbf{\Delta})$ is the imaginary part of the
dipole-proton elastic scattering amplitude. The energy dependence of the latter
is via the dimensionless variable $x$ taken here to
be
\begin{equation}
x = (Q^2 + 4m_f^2)/(Q^2+ s)
\end{equation}
where $m_f$ is a phenomenological light quark mass.\footnote{Here we shall take
$m_f=0.14~\mbox{GeV}$ i.e the
value used when extracting the dipole
cross-section from $F_2$ data.} Setting $t=0$ in equation
\eqref{non-forward-amplitude}, we obtain the forward
amplitude used in reference~\cite{Forshaw:2010py}, i.e
\begin{equation}
\left.\Im \mbox{m} \, \mathcal{A}_\lambda(s,t;Q^2)\right|_{t=0} = s \sum_{h,
\bar{h}}
\int \mathrm{d}^2 {\mathbf r} \; \mathrm{d} z \; \Psi^{\gamma,\lambda}_{h, \bar{h}}(r,z;Q^2)
\hat{\sigma}(x,r) \Psi^{\rho,\lambda}_{h, \bar{h}}(r,z)^*
\label{forward-amplitude}
\end{equation}
where we have used the optical theorem to introduce the dipole cross-section
\begin{equation}
\hat{\sigma}(x,r)=\frac{\mathcal{N}(x,r,\mathbf{0})}{s}\;.
\label{dipole-xsec-optical}
\end{equation}
Note that since the momentum transfer $\mathbf{\Delta}$ is Fourier conjugate to
the impact parameter $\mathbf{b}$, the dipole cross-section at a given energy
is simply the $b$-integrated dipole-proton scattering amplitude, i.e
\begin{equation}
\hat{\sigma}(x,r)=\frac{1}{s}\int
\mathrm{d}^2 \mathbf{b} \mathcal{N}(x,r,\mathbf{b}) \;.
\label{dipole-xsec-b-int}
\end{equation}
This dipole
cross-section can be extracted from
the $F_2$ data since
\begin{equation}
F_2(x,Q^2)  \propto \int \mathrm{d}^2 \mathbf{r} \mathrm{d} z |\Psi_{\gamma^*}(r,z;Q^2)|^2
\hat{\sigma}(x,r)
\end{equation}
and the photon's light-cone wavefunctions are known in QED, at least for large
$Q^2$. The
$F_2$-constrained dipole cross-section can then be used
to predict the imaginary part of the forward amplitude
for diffractive $\rho$ production and thus the forward differential
cross-section,
\begin{equation}
\left. \frac{d\sigma_{\lambda}}{dt} \right|_{t=0} =\frac
{1}{16\pi} (\Im\mathrm{m} \mathcal{A}_\lambda(s,0))^2 \; (1 + \beta_\lambda^2)~,
\label{gammap-xsec}
\end{equation}
where $\beta_\lambda$ is the ratio of real to imaginary parts of the
amplitude and is computed as in reference~\cite{Forshaw:2010py}. The
$t$-dependence
can be assumed to be the
exponential dependence as suggested by experiment~\cite{Chekanov:2007zr}:
\begin{equation}
\frac{d\sigma_{\lambda}}{dt}= \left. \frac{d\sigma_{\lambda}}{dt} \right|_{t=0}
\times
\exp(-B|t|)
\label{exponential-t}
\end{equation}
where
\begin{equation}
B=N\left(
  14.0 \left(\frac{1~\mathrm{GeV}^2}{Q^2 + M_{\rho}^2}\right)^{0.2}+1\right)
\label{Bslope}
\end{equation}
with $N=0.55$ GeV$^{-2}$. After integrating over $t$, we can compute the total
cross-section $\sigma=\sigma_L + \epsilon \sigma_T$
which is measured at HERA.\footnote{To compare with the HERA data, we take
$\epsilon=0.98$.}

Presently several dipole
models~\cite{Forshaw:2004vv,Watt:2007nr,Soyez:2007kg,Bartels:2002cj,
Kowalski:2006hc}
are able to fit the current HERA $F_2$ data and
there is evidence that the data prefer those incorporating
some form of saturation~\cite{Motyka:2008jk}. We can use the $F_2$-constrained
dipole cross-section in order to
extract the $\rho$ light-cone wavefunction using the current precise HERA data
~\cite{Chekanov:2007zr,Aaron:2009xp}. This
has recently been done in reference~\cite{Forshaw:2010py} using the
Regge-inspired  FSSat dipole
model~\cite{Forshaw:2004vv} and we shall report the results of
this work here. In addition, we repeat the analysis
using two alternative models~\cite{Soyez:2007kg,Watt:2007nr,Kowalski:2006hc}
both
based on the original Colour Glass Condensate (CGC) model~\cite{Iancu:2003ge}.
They differ from the original CGC model by including the
contribution of charm quarks when fitting to the $F_2$
data. Furthermore in one of them~\cite{Soyez:2007kg,Watt:2007nr}, the anomalous
dimension  $\gamma_s$ is treated as an additional free parameter instead of
being fixed to its LO BFKL value of $0.63$. We shall refer to
these models as CGC[$0.74$] and CGC[$0.63$] models  where the number in the
square brackets stands for the fitted and fixed value of the anomalous
dimension respectively. For both models, we use the set of fitted parameters
given in reference~\cite{Watt:2007nr}. All
three models, i.e FSSat, CGC[$0.63$] and CGC[$0.74$] account for saturation
although in a $b$- (or equivalently $t$-) independent way. Indeed, at
a given energy, the dipole cross-section is equal to the forward
dipole-proton amplitude given by equation \eqref{dipole-xsec-optical} or to the
$b$-integrated
dipole proton amplitude given by equation \eqref{dipole-xsec-b-int}. Finally
we note that all three dipole models we consider here give a good description of
the diffractive structure function, i.e $F_2^{D(3)}$
data~\cite{Marquet:2007nf,Forshaw:2006np}.

\section{Fitting the HERA data}

Previous work~\cite{Forshaw:2003ki,Marquet:2007qa,Watt:2007nr}
has shown that a reasonable assumption for the
scalar part of the light-cone wavefunction for the $\rho$ is of the form
\begin{eqnarray}
\phi^{{\mathrm{BG}}}_\lambda(r,z) &=&
\mathcal{N}_\lambda \;  4[z(1-z)]^{b_{\lambda}} \sqrt{2\pi R_{\lambda}^{2}} \;
\exp \left(\frac{m_f^{2}R_{\lambda}^{2}}{2}\right)
\exp \left(-\frac{m_f^{2}R_{\lambda}^{2}}{8[z(1-z)]^{b_{\lambda}}}\right) \\
\nonumber
& &\times \exp \left(-\frac{2[z(1-z)]^{b_\lambda}
r^{2}}{R_{\lambda}^{2}}\right)
\label{boosted-gaussian}
\end{eqnarray}
and is referred to as the 'Boosted Gaussian' (BG). This wavefunction is a
simplified version of that proposed originally
by Nemchik, Nikolaev, Predazzi and Zakharov~\cite{Nemchik:1996cw}. In the
original BG wavefunction, $b_\lambda=1$ while the parameters $R_{\lambda}$ and
$\mathcal{N}_{\lambda}$ are fixed by the leptonic decay width constraint and
the wavefunction normalization conditions~\cite{Forshaw:2010py}.

However, when the BG wavefunction is used in conjunction with either the FSSat
model or any of the CGC models, none of them is able to give a good quantitative
agreement with the current HERA $\rho$-production data.
This is illustrated by the large $\chi^2$ values in table \ref{tab:BG-chisqs}.
As shown in table \ref{tab:BG-Fits-chisqs}, this
situation is considerably improved by fitting $R_\lambda$ and $b_\lambda$ to
the leptonic decay width and HERA data.\footnote{We fit to the same data set and
with the same cuts as in reference~\cite{Forshaw:2010py}.}

\begin{table}[h]
\begin{center}
\textbf{Boosted Gaussian predictions}
\[
\begin{array}
[c]{|c|c|}\hline
\mbox{Dipole model} & \chi^2/\mbox{data point}\\ \hline
\mbox{FSSat}& 310/75 \\ \hline
\mbox{CGC}[0.74]& 262/75 \\ \hline
\mbox{CGC}[0.63]&  401/75 \\ \hline
\end{array}
\]
\end{center}
\caption {Predictions of the $\chi^2/\mbox{data point}$ using the BG
wavefunction.}
\label{tab:BG-chisqs}
\end{table}

\begin{table}[h]
\begin{center}
\textbf{BG fits}
\[
\begin{array}
[c]{|c|c|}\hline
\mbox{Model} & \chi^2/\mbox{d.o.f}  \\ \hline
\mbox{FSSat~\cite{Forshaw:2010py}}&  82/72 \\ \hline
\mbox{CGC}[0.74]& 64/72 \\ \hline
\mbox{CGC}[0.63] & 83/72 \\ \hline
\end{array}
\]
\end{center}
\caption {$\chi^2/\mbox{d.o.f}$ obtained when fitting $R_{\lambda}$ and
$b_{\lambda}$ to the leptonic decay width and HERA data.}
\label{tab:BG-Fits-chisqs}
\end{table}

For the FSSat and CGC[$0.63$] models, we can further improve the quality of fit
by allowing for
additional end-point enhancement in the transverse wavefunction, i.e. using a
scalar
wavefunction of the form
\begin{equation}
\phi_T (r,z)= \phi^{{\mathrm{BG}}}_T (r,z) \times [1+ c_{T}
\xi^2 + d_{T} \xi^4]
\label{EG}
\end{equation}
where $\xi=2z-1$. The results are shown in table
\ref{tab:Enhancement-fits-chisqs}.

\begin{table}[h]
\begin{center}
\textbf{Improved fits}
\[
\begin{array}
[c]{|c|c|}\hline
\mbox{Model} & \chi^2/\mbox{d.o.f} \\ \hline
\mbox{FSSat~\cite{Forshaw:2010py}}&  68/70 \\ \hline
\mbox{CGC}[0.63] & 67/70  \\ \hline
\end{array}
\]
\end{center}
\caption {$\chi^2/\mbox{d.o.f}$ obtained when fitting $b_{\lambda}$,
$R_{\lambda}$
$c_{T}$, $d_{T}$ the leptonic decay width and HERA data.}
\label{tab:Enhancement-fits-chisqs}
\end{table}

\begin{table}[h]
\begin{center}
\textbf{Best fit parameters}
\[
\begin{array}
[c]{|c|c|c|c|c|c|c|}\hline
                 & R_L^2 & R_T^2 & b_L & b_T & c_T & d_T \\ \hline
\mbox{FSSat~\cite{Forshaw:2010py}}     &26.76  &27.52 &0.5665 &0.7468 &0.3317
&1.310  \\ \hline
\mbox{CGC}[0.63] &27.31  &31.92 &0.5522 &0.7289 &1.6927 &2.1457  \\ \hline
\mbox{CGC}[0.74] &26.67  &21.30 &0.5697 &0.7929  &0&0 \\ \hline
\end{array}
\]
\end{center}
\caption {Best fit parameters for each dipole model.}
\label{tab:Best-fit-params}
\end{table}

\begin{figure}
\begin{center}
\includegraphics[width=0.8\textwidth]{H1xsec.pdf}
\end{center}
\caption{\label{fig:H1-xsec} Best fits to the HERA total cross-section data.
CGC[$0.74$]: solid;
FSSat: dotted; CGC[$0.63$]: dashed.}
\end{figure}

\begin{figure}
\begin{center}
\includegraphics[width=0.8\textwidth]{ZEUSxsec.pdf}
\end{center}
\caption{\label{fig:ZEUS-xsec} Best fits to the ZEUS total cross-section data.
CGC[$0.74$]: solid;
FSSat: dotted; CGC[$0.63$]: dashed.}
\end{figure}

\begin{figure}
\begin{center}
\includegraphics[width=0.4\textwidth]{H1ratio.pdf}
\includegraphics[width=0.4\textwidth]{ZEUSratio.pdf}
\end{center}
\caption{\label{fig:HERA-ratio} Best fits to the $\sigma_L/\sigma_T$ data. The
H1 data are at $W=75$~GeV while the ZEUS data are at $W=90$~GeV.
CGC[$0.74$]: solid;
FSSat: dotted; CGC[$0.63$]: dashed.}
\end{figure}

The best fits obtained with each dipole model are compared to the HERA data in
figure \ref{fig:H1-xsec}, \ref{fig:ZEUS-xsec} and \ref{fig:HERA-ratio}.
The corresponding fitted parameters are given in table
\ref{tab:Best-fit-params}.
Note that we achieve a lower $\chi^2/\mbox{d.o.f}=0.89$ with CGC[$0.74$]
than with CGC[$0.63$] and FSSat for which we obtain $\chi^2/\mbox{d.o.f}=0.96$
and $\chi^2/\mbox{d.o.f}=0.97$ respectively. Compared to
the FSSat and CGC[$0.63$] fits, note that no
additional enhancement in the transverse wavefunction is required in the
CGC[$0.74$] fit. Nevertheless the
extracted wavefunction still exhibits enhancement compared to the old BG
wavefunction. The extracted light-cone wavefunctions are shown in figure
\ref{fig:Psisq_r0}.

\begin{figure}
\begin{center}
\includegraphics[width=0.4\textwidth]{Psisq_L_r0.jpeg}
\includegraphics[width=0.4\textwidth]{Psisq_T_r0.jpeg}
\end{center}
\caption{\label{fig:Psisq_r0}The longitudinal (left) and transverse (right)
light-cone
wavefunctions squared at $r=0$. CGC[$0.74$]: solid; FSSat: dotted; CGC[$0.63$]:
dashed.}
\end{figure}

\section{Distribution Amplitudes}
The leading twist-$2$ Distribution Amplitude (DA) is given by
~\cite{Forshaw:2010py}
\begin{equation}
\varphi(z,\mu) \sim \left( 1 - \mathrm{e}^{-\mu^2/\Delta(z)^2}\right)
  \mathrm{e}^{-m_f^{2}/\Delta(z)^2} [z(1-z)]^{b_L}~,
\end{equation}
where $\Delta(z)^2 = 8[z(1-z)]^{b_L}/R_L^2$. This leading twist DA is only
sensitive to the longitudinal wavefunction and, as illustrated in figure
\ref{fig:DA_1GeV}, we expect little variation in the predictions using the
different dipole models.
To compare with existing theoretical predictions for the DA, we
compute moments, i.e.
\begin{equation}
\langle \xi^n \rangle_{\mu} = \int_0^1 \mathrm{d} z \; \xi^n \varphi(z,\mu)~.
\end{equation}
where by convention~\cite{Forshaw:2010py}
\begin{equation}
\int_0^1 \mathrm{d} z ~\varphi(z,\mu) = 1~.
\end{equation}

In reference~\cite{Forshaw:2010py}, we noted that our DA is very slowly varying
with
$\mu$ for $\mu > 1$~GeV, i.e  our parameterization neglects the
perturbatively known
$\mu$-dependence of the DA. This statement remains true
if we use the CGC[$0.63$] or CGC[$0.74$] instead of the FSSat model.

Our results are compared with the existing predictions in table
\ref{tab:moments-mu}. The moments
obtained with our best fit, i.e with the CGC[$0.74$] model, are very similar to
those obtained with FSSat model or the CGC[$0.63$]. In all cases, the results
are in very good agreement with expectations based on QCD sum rules and the
lattice.

\begin{table}[h]
\begin{center}
\textbf{Moments of the leading twist DA at the scale $\mu$}
\[
\begin{array}
[c]{|c|c|c|c|c|c|c|c|c}\hline
\mbox{Reference} & \mbox{Approach} & \mbox{Scale}~\mu &\langle \xi^2
\rangle_{\mu}&\langle \xi^4 \rangle_{\mu}&\langle \xi^6 \rangle_{\mu}&\langle
\xi^8 \rangle_{\mu}&\langle \xi^{10} \rangle_{\mu} \\ \hline
\mbox{(This paper)} & \mbox{CGC[$0.74$] fit}&\sim 1~\mbox{GeV} &0.227 &
0.105& 0.062 &0.041 &0.029\\ \hline
\mbox{(This paper)} & \mbox{CGC[$0.63$] fit}&\sim 1~\mbox{GeV} &0.229 &
0.107& 0.063 &0.042 &0.030 \\ \hline
\mbox{\cite{Forshaw:2010py}} & \mbox{FSSat fit}&\sim 1~\mbox{GeV} &0.227&
0.105&0.062&0.041&0.029\\ \hline
\mbox{(This paper)} & \mbox{Old BG prediction}&\sim 1~\mbox{GeV} &0.181&
0.071&0.036&0.021&0.014\\ \hline
\mbox{\cite{Bakulev:1998pf}}&
\mbox{GenSR}&1~\mbox{GeV}&0.227(7)&0.095(5)&0.051(4)&0.030(2)&0.020(5) \\ \hline
\mbox{\cite{Chernyak:1983ej}}& \mbox{SR}&1~\mbox{GeV} &0.26&0.15 & & &  \\
\hline
\mbox{\cite{Ball:1996tb}} & \mbox{SR}&1~\mbox{GeV} &0.26(4)& & & &   \\ \hline
\mbox{\cite{Ball:2007zt}}&\mbox{SR}&1~\mbox{GeV} &0.254& & & &  \\ \hline
\mbox{\cite{Ball:2004ye}}&\mbox{SR}&1~\mbox{GeV} &0.23\pm^{0.03}_{0.02}&
0.11\pm_{0.02}^{0.03}& & &  \\ \hline
\mbox{\cite{Boyle:2008nj}}&\mbox{Lattice} &2~\mbox{GeV} &0.24(4)& & & &  \\
\hline
 & 6z(1-z)
&\infty&0.2&0.086&0.048&0.030&0.021 \\ \hline
\end{array}
\]
\end{center}
\caption {Our extracted values for  $\langle \xi^n \rangle_{\mu}$, compared to
predictions based on the QCD sum rules (SR),
Generalised QCD Sum Rules (GenSR) or lattice QCD.}
\label{tab:moments-mu}
\end{table}

Finally, in figure \ref{fig:DA_1GeV} we compare our DAs with that
predicted by Ball and Braun~\cite{Ball:1996tb}, at a scale $\mu=1$ GeV.
The agreement is reasonable given that in reference~\cite{Ball:1996tb}, the
expansion
in Gegenbauer polynomials is truncated at low order, which is presumably
responsible for the local minimum at $z=1/2$. Certainly all $4$ distributions
distributions are broader than the asymptotic prediction $\sim 6z(1-z)$.

\begin{figure}
\begin{center}
\includegraphics[width=0.4\textwidth]{DAL.jpeg}
\end{center}
\caption{\label{fig:DA_1GeV}The extracted leading twist-$2$ DAs at $\mu=1$ GeV
(CGC[$0.74$]: solid;
FSSat: dotted; CGC[$0.63$]: dashed) compared to the DA of reference
~\cite{Ball:1996tb} also at $1$ GeV (long-dashed) and the asymptotic
DA (dot-dashed).}
\end{figure}

\section{Conclusions}
We have used the current HERA data on diffractive $\rho$ production to extract
information on the $\rho$ light-cone wavefunction. We find that the
corresponding leading twist-$2$ DA is broader than the asymptotic shape
and agrees very well with the expectations of QCD sum
rules and the lattice. We also find that the data
prefer a transverse wavefunction with end-point enhancement although the degree
of such an enhancement is model-dependent.

\section*{Acknowlegdements}

R.S thanks the Institute for Nuclear Theory (INT) and the Facult\'e des \'Etudes
Superieures et de la Recherche (FESR) of the Universit\'e de Moncton for
financial support. R.S also thanks the organisers for their invitation and for
making this workshop most enjoyable. We thank H. Kowalski and C.
Marquet for useful discussions. This research is also supported by the
UK's STFC.

redpajama_set_name: RedPajamaArXiv


text:
Richard F Lacaille
Global Chief Investment Officer
Rick Lacaille, Executive Vice President, became Global Chief Investment Officer (CIO) of State Street Global Advisors in October 2008 and is a member of the firm's Executive Management Group. In his role as Chief Investment Officer Rick has responsibility for all investment management activity at State Street Global Advisors, including research and trading.
Prior to his current role, Rick was Head of Global Active Equities, and previously European CIO. Before joining State Street Global Advisors in 2000, he held a wide variety of posts in quantitative fund management and research at Gartmore Investment Management, including periods as Head of Quantitative Research and Head of Structured Equities.
Rick has a BSc (Hons) in Operational Research from Lancaster University and MSc in Econometrics from London Guildhall University.
Rick is a member of the FTSE Russell Policy Advisory Board, the MSCI Editorial Advisory Board and the Asset Management Committee of the Investment Association. He is a regular writer and broadcaster on investment issues and speaks frequently at industry conferences.