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Greeba Barlow
Nancy Greco, Features Editor|December 1, 2005
Like many seniors, Greeba Barlow has high expectations for her last year at St. John's. The 5-foot-10 guard is hoping to make her final year on the basketball team her most successful one.
Barlow had an impressive career before coming to St. John's. During her high school years in Columbia, Md., she was a member of the Adidas Top 10 All-Star team.
Then she attended George Washington University, where she played in 30 of 31 games and participated in the 2003 NCAA Tournament's first and second rounds.
She was also a member of the 2003 Atlantic 10 Championship team before she decided that she needed a change. She felt that being in the Big East would provide her with more of a challenge, which was something she welcomed.
"I felt I needed a new start," Barlow said.
She transferred to St. John's and sat out the 2003-04 season to fulfill NCAA transfer requirements.
St. John's has a different dynamic than George Washington, and Barlow likes being part of the up and coming program.
"We had true seniors [at George Washington] that were there for four years. I had to prove myself," she said. "Here, there is no set of players who have been here and can look after the freshmen. It's not a set structure because they're building the team."
As a junior at St. John's, Barlow appeared in 29 games and averaged 21.3 minutes, 8.3 points and 2.3 rebounds. On Feb. 6, 2005, she was named Big East Player of the Week, becoming the first St. John's player to earn this distinction under coach Kim Barnes Arico's direction.
She also appreciates the ambition that Barnes Arico demonstrates with the team.
"She's a great person," Barlow said. "She is trying to get the program to be a top Big East program."
While Barlow considers her biggest strength to be in her shooting, she hopes to work this season to improve her defense and her own attitude toward the game.
"The challenge for me is myself," Barlow said. "I have to overcome myself in the mental part."
As a senior, Barlow is helping the team develop a closer relationship. She has become someone the younger players on the team can look up to.
"I think we have a good chemistry, but we're working on it," she said. "Eventually, it's going to come together."
Barlow hopes that if she performs well this season, she may have a chance to play for the WNBA or overseas in the future.
"I think she's shown us she wants to be successful at St. John's and in the Big East," Barnes Arico said. "I think if she stays healthy, she can have a really successful senior year."


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17/04/19<br>Aston Martin Unveil Rapide E Electric Car
Aston Martin Unveil Rapide E Electric Car
Following a stunning concept car in 2015 and an official announcement back in 2017, Aston Martin's very first all-electric car is officially here. Named the Aston Martin Rapide E, this limited run production car is the first in a future evolution of the brand's range.
At first glance, it's easy to think that the Rapide E is just an electric conversion of the existing Aston Martin Rapide, it isn't. Developed in collaboration with Williams Advanced Engineering, Rapide E merely borrows the chassis and beautiful styling of the Rapide and that's about it.
The most notable of changes is probably the fact there's no longer a naturally aspirated 6.0-litre V12 under that long bonnet; instead, there's now an all-new battery pack encased in a carbon fibre and Kevlar casing.
Utilising over 5,000 lithium-ion cells, that battery pack boasts 800V and 65kWh and is apparently better than other electrical systems when it comes to charging and thermal management.
That bespoke electrical system, built alongside Williams Advanced Engineering, grants the Rapide E the equivalent of 601bhp and 700lb ft of torque. That then translates into a 0-60mph sprint of under four seconds, a 50-70mph sprint of 1.5 seconds and a top speed of 155mph. Impressive, no?
What's even more impressive is that Aston Martin says Rapide E will be able to accomplish these feats consistently, time-and-time again. In fact, Aston Martin are so confident that their new battery pack never loses efficiency that they've been quoted saying that it'll do an entire lap of the infamous Nürburgring without ever suffering any performance derating.
Despite this focus on performance, the range that Rapide E can cover hasn't been compromised. That means an estimated WLTP cycle range of over 200 miles and, thanks to the 800V high-voltage battery system, the ability to connect to 100kW chargers for fast charging – 310 miles every hour. There's also a high-powered AC on-board charger capable of recharging the battery in less than 3 hours when out-and-about.
Of course, the Rapide E is an Aston Martin, meaning that it needs to be more than just powerful, it also needs to drive well. Ensuring that it retains the great driving characteristics of the Rapide, Aston Martin have equipped Rapide E with rear-wheel drive, a limited-slip differential, revised spring and damper rates, a redesigned underfloor and a bigger rear diffuser; the latter two helping to improve aerodynamics.
Dr.Andy Palmer, Aston Martin CEO, said:
"As our first all-electric production car, it [Rapide E] is a truly historic step. As a car company we cannot afford to passively allow that future to come to us; we have to actively chase it.
Only by doing this can we learn and prepare, but also preserve those things we love as drivers and car enthusiasts. That's why it was so important to me that in embracing EV technology we should not let go of those unique qualities that define an Aston Martin. I believe Rapide E embodies that desire and paves the way for a hugely exciting future."
Production of the all-new Aston Martin Rapide E will be strictly limited to just 155 examples and will take place at their state-of-the-art St Athan facility – a place otherwise known as the brand's Home of Electrification. If you find yourself fancying one of your own, then it's available to order now but price is available on application only.
What do you think of the stunning new Aston Martin Rapide E? We'd love to read your thoughts on Facebook, Instagram or Linkedin.
UK to Reduce National Grid Strain with Smart Charging Pilot
Could localised electricity storage hubs minimise strain on the national grid?
Hybrid and Electric Vehicles at the Sytner Group
Have questions about hybrid and electric cars? Take a look at our helpful new page!
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redpajama_set_name: RedPajamaCommonCrawl


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text:
Menotti, Gian Carlo (1911-2007)
Ensemble with Photograph and Autograph Musical Quotation Signed
"Gian Carlo Menotti". 1972. Decoratively matted in silver and with a silver frame. Total size: 16.25" h x 19.75" w (41.3 cm x 50.2 cm). Excellent condition.
A framed ensemble with a photograph of the noted Italian-American composer and librettist Gian Carlo Menotti and a signed cover of his one-act comic opera The Telephone. Schirmer published the music, using for the cover a design by Saul Steinberg, renowned for his covers and drawings that appeared in The New Yorker for nearly six decades. Menotti has signed the cover underneath a one-bar quotation from the work with his hand-written staff, below which he has written the lyrics Hallo! Hello!.... Dated by him at the bottom of the cover.
The Telephone, or L'Amour à Trois, is an English-language comic opera in one act. Menotti wrote both the words and the music in 1945 and the work premiered in New York City on February 18, 1947. A German film was made of it in 1968. It is one of several operas for which Menotti is best remembered; among the others are The Medium, The Consul, Amahl and the Night Visitors, and The Saint of Bleeker Street.


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Election Commentary
This Ohio magistrate came out as trans at work. Now she's getting community support to be a judge.
When Millas introduced herself to her coworkers as Ginny for the first time, she was met with flowers, cards, and an outpouring of compassion.
Commentary by Becca King Tuesday, March 17, 2020
Ohio judge's magistrate Ginny MillasPhoto: provided
For Ginny Millas, coming out at work meant making history, opening the doors of opportunity for herself and for others.
Millas is the first transgender judge's magistrate in the state of Ohio and is now running for the office of Cuyahoga County Domestic Relations Court judge. She would be the first transgender person to hold a superior court judgeship in the state of Ohio if elected, and one of few transgender people to hold such an office nationally.
Related: Elder trans comedian slays judges on America's Got Talent
Millas has worked as a Custody Magistrate for the Cuyahoga County Juvenile Court for the past seven years, helping thousands of families through custody battles, often involving difficult cases of abuse and neglect. She speaks of decisions to separate families gravely, preferring wherever possible to rehabilitate, assist and educate parents.
Previously, Millas has worked as a prosecutor and as a public defender, with 25 years in total spent as a public servant.
Holding the office of Domestic Relations Court judge would mean continuing to make decisions about custody, as well as divorce, intimate violence, and other domestic conflicts.
Millas also sees the judgeship as an opportunity to be a community leader who can speak to issues of injustice more broadly and affect the public's expectations of transgender people. She hopes to clarify why for her and for so many others a transition is necessary.
She describes life before transitioning as "like being buried alive in a coffin," a kind of suffocation with no inner peace. Something felt deeply wrong, though she couldn't identify what it was exactly until later in life. She only knew that she hated herself, hated her body, and for some reason felt stunted in her ability to express her emotions.
Realizing that she could live as a woman "was emancipating beyond description," and the relief she felt from hormone therapy was "stupendous." Though her marriage ended as a result of the changes, with the work of a therapist, she's been able to reinvent herself and find true self-love for the first time.
"I just really blossomed," said Millas.
Still, being out in the courtroom was a terrifying concept. In many parts of the state, a gender transition on the job can even be grounds for firing. Only after four years of being out in her personal life and receiving heavy encouragement from friends did she summon the courage to break that barrier.
"You deserve to be yourself at work," a close friend said.
"I could rationalize why our county wasn't ready for that, and the law field was too conservative," said Millas. "But that, that stunned me, because there wasn't an answer for that. I realized she was right. And I created this big plan about how I would go about doing this."
The plan involved setting up a meeting with her supervisor Judge Michael Ryan, along with representatives from Human Resources. She came out to them first, then discussed how she could best share the news with her colleagues.
The court employs thousands of people, making this no small undertaking. But her supervisor and HR representatives offered her support and encouragement far beyond what she'd expected.
"You're going to be better, Ginny," she remembers Judge Ryan saying. "You're going to be more relaxed. You're going to be more yourself."
"I was so impressed, he got it in such a profound way," said Millas.
Ultimately, they crafted a letter to all the court's employees signed by each judge, explaining her transition and notifying all that she would be presenting as female in the near future following a two-week vacation.
When Millas introduced herself to her coworkers as Ginny for the first time, she was met with flowers, cards, and an outpouring of compassion. If any colleagues did have transphobic comments, they kept them to themselves.
"The people with malice were few and far between," said Millas.
Millas finds inspiration in her Jewish faith, likening the transgender experience to the story of Exodus. She says that escaping slavery in Egypt was only the first step and that it was followed by a long, multigenerational struggle towards the safety and security of a free nation.
"You have to go forward with courage and conviction that your voice needs to be heard, and that the outcome isn't predetermined."
She describes her mom as one of her strongest advocates, saying that her support has made the whole process possible. Though her father passed before she could share the news with him, she describes vivid dreams she's had working through it with him and feels strongly about the impact that supportive parents have on their children's lives, even into adulthood.
To be a judge would be a dream come true, and one Millas is willing to work towards for years if necessary.
"I'll just do the best I can this cycle and observe and watch how people are doing it who have done it for over a year now," said Millas about campaigning. "The sheer volume of people that I've encountered over my career is really a lot. And so if I do well, I will attribute quite a lot to just the reputation I've developed over the years."
Ginny Millas, Good News, judge magistrate, judicial election, Ohio, trans candidates
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This is not simply a time for condemnation. Those who were horrified by what we witnessed on January 6 must take action.


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The Old Vicarage , Sharnbrook, Bedfordshire, 1870
inscribed and dated in the margin " Sharnbrook Vicarage, Bedfordshire, 1870"
13 x 17.50 cm.
Sharnbrook village is in the west of the parish. The church of St. Peter is situated on the west of the road as it approaches the village. The old vicarage, which was pulled down about 1910, stood about 180 yds. from the present new house. It was built of stone with a thatched roof, and was an interesting specimen of a priest's house of the 16th century.
Sharnbrook is a parish 9 miles north-west from Bedford. It covers an area of 2,467 acres, of which 853 acres are arable land and 1,305¾ acres permanent grass. (fn. 2) The parish is watered by the River Ouse, which forms the southern boundary. The slope of the ground is from north to south, the highest point above ordnance datum is 303 ft. in the north, the lowest 143 ft. in the south. The soil is clay, the subsoil clay and gravel, the principal crops being wheat, barley, oats, peas, beans and turnips.
The Tofte, a small but substantially built stone mansion, occupies a pleasant position about half a mile northward of the church. In the gable on the south front is an inscription 'Except the Lord build the house their labour is but lost that build it. 1613.' It has been much modernized and a new wing erected in recent years. Whilst taking down the walls during the alterations various pieces of old masonry were discovered. Forty feet to the north of the old house some old buttressed foundations were found. In 1907 human remains were found in the grounds about 3 ft. below the surface. It marks the site of a manor formerly belonging to Newnham Priory, and is at present the property of and occupied by Mr. A. R. Alston. East of the church is Sharnbrook House, in the occupation of Mr. E. Arkwright, Master of the Oakley Hunt. It is a Georgian house of red brick, with stone quoins and a rich wood cornice, and was formerly known as Towns End House.
Sharnbrook Grange, a 17th-century house, stands in park-like grounds of 25 acres. It is a rectangular brick building with a tile roof. Through the cellar is a constant flow of water, but whence it comes is not known. In the garden are some fine cedars and two stone basins dated 1658 and 1745 respectively.
The Midland Railway, which has a station at the east end of the village, has a bridge of ten arches across the Ouse.
Colworth, which is described as a hamlet of Sharnbrook in 1278–9, now consists principally of the Colworth House estate. This house, which was built in the 18th century by Mark Antonie, stands in a well-wooded park of 176 acres. Lysons describes it as one of the most elegant mansions in the county. (fn. 3) It is at present the residence of Mr. Albert Edward Bowen. Sharnbrook is well wooded, containing in all 269¼ acres. (fn. 4) Among the woods may be named Halsey Wood, with the largest area, Francroft Wood, Round Wood, Woodend Plant and the Swell, the last-named being in the Colworth park.
There is a small industry in hand-made lace, dating from the 16th century, still carried on in this parish.
Two mills still exist in Sharnbrook, one a disused windmill, in the south, the second a steam corn-mill on the site of a former water-mill in the south-west near the approach to the village.
There are two Baptist chapels in Sharnbrook.
The following place-names have been found in documents connected with this parish:—Carswellfielde, Sondewood, Temple Leys, Temple Corner (xvi cent.), Castle Close, Coffle End, Clipsey Spinney, Eyles Plantation, Naboth's Vineyard, Uttersfield Plantation (xx cent.).
At the Survey of 1086 eight proprietors of land are named as holding in Sharnbrook. First amongst these Domesday tenants may be mentioned Albert of Lorraine, whose lands subsequently became known as LORINGS or PIPINGS MANOR. In 1086 Albert of Lorraine, who had been preceded by Algar, a man of Queen Edith, held 2 hides, a quarter of a virgate and a mill worth 16s. (fn. 5) He owned land, including a 10-hide manor at Wootton, elsewhere in Bedfordshire, and, as has been shown by Mr. Round, (fn. 6) was a clerk or chaplain who enjoyed the favour alike of Edward the Confessor and of William, and received from them lands and houses.
With regard to the overlordship of this manor, it was held by Albert of Lorraine of the king in chief in 1086, (fn. 7) but in 1185 is found attached to the barony of Bedford, (fn. 8) to which overlordship the latest reference has been found in 1419. (fn. 9)
The reasons for connecting the Loring family, who subsequently held in this parish, with Albert of Lorraine, the clerk of Domesday, may be briefly summarized as follows:—Albert held in four Bedfordshire parishes at Domesday: Chalgrave, Wootton, Shelton, Sharnbrook. Some century and a half later it is found that in three out of these four, Chalgrave, Wootton and Sharnbrook, the Loring family are holding by knight service, and that moreover all three have become attached to the barony of Bedford. (fn. 10) The chief seat of the Lorings was at Chalgrave; the descent of their Sharnbrook Manor was as follows:—Roger Loring died some time before 1185, at which date his son Peter, aged twelve, was in the guardianship of the king. (fn. 11) This Peter was still alive in 1250, in which year he was defeated in a suit against Dunstable Priory, (fn. 12) and it was probably his son Peter who died in 1286, and whose epitaph is given by the Dunstable chronicler as 'qui super omnes progenitores suos specialius nos amavit.' (fn. 13) The feudal assessment for 1302 gives Peter Loring as still holding one-fifth fee in Sharnbrook. (fn. 14) Roger Loring his son died before 1346, (fn. 15) and was succeeded by his son the eminent Sir Nigel Loring, a founder Knight of the Garter, who on his death in 1385–6 left two daughters as co-heirs. Of these Isabel became the wife of Robert Lord Harington, (fn. 16) and acquired Sharnbrook as part of her share in her father's property. Her husband died in 1406, when the Sharnbrook property included a messuage, 100 acres of land, 20 acres meadow and 10 acres of wood. (fn. 17) Their son John Lord Harington died in 1417–18 without issue, and his brother William succeeded him. (fn. 18) In 1449 William Lord Harington conveyed this estate, called for the first time a manor, into the hands of the Archbishop of Canterbury and other trustees. (fn. 19) He died in 1458, leaving as heir Elizabeth wife of William Bonville. (fn. 20) Her son William, who acquired the title Lord Harington in right of his mother, was slain at Wakefield in 1460, and Sharnbrook passed to his daughter Cecilia, who became the wife of Thomas Grey first Marquess of Dorset.
Newnham Priory owned TOFTS or THE TOFTE MANOR in Sharnbrook from the 13th century. Its origin is to be sought to a great extent in the land which Count Eustace held at Domesday, (fn. 34) further supplemented by grants from the Beauchamp fee. Robert son of Rozelin, who was Count Eustace's tenant in 1086, was succeeded by the Trikets, of whom four generations are mentioned in Newnham Cartulary as benefactors of the priory in the 12th and 13th centuries—William son of Simon Triket, Baldwin son of William and Simon son of Baldwin. (fn. 35) Their gifts, which were all held of the honour of Boulogne (to which Count Eustace's lands subsequently became attached), ranged from 2 virgates of land in Toft—whence the manor derived its distinctive name—to 3 acres of land in Tranecroft, and by 1278–9 included a capital messuage, 180 acres of land, meadow and pasture, and 10 acres of wood. (fn. 36) Newnham Priory also received various grants of land from William Druel and other members of the same family, who held of the barony of Bedford, 13 selions from Robert de Parentine, and 27 selions from John le Child, whose father also granted the priory certain inclosed waters 'from Sitegate to Besebrigg.' (fn. 37)
Newnham Priory therefore held land in Sharnbrook attached to these two honours of Boulogne and Bedford. In 1281 the prior protested against appearing as suitor at the court of the former honour, saying that he owed no service though often distrained by the bailiffs. (fn. 38) The prior's name appears, however, in 1302 as holding a half fee of this honour, (fn. 39) whilst in 1346 and again in 1428 he held half a fee of the barony of Bedford. (fn. 40)
The manor was in lease to William Tofte in 1331 by assignment from one Bartholomew and Ellen his wife, who held it for their lives. (fn. 41) In 1385 the prior received a charter of free warren in Sharnbrook, (fn. 42) and at the Dissolution this manor became Crown property. (fn. 43) In 1540 George Boteler received a grant of this manor, (fn. 44) which henceforward follows the same descent as Loring s (q.v.). The Tofte has long been the residence of the lords of the manor and the principal house in the village.
¶A third manor in Sharnbrook, that of TEMPLE HILLS, was owned from the 12th century by the Knights Templars. The Bishop of Coutances held in all 4½ hides in Sharnbrook in 1086. This land, in which he had been preceded by Borred, was held of him by various tenants. Turgis an Englishman held half a hide, Humphrey held half a hide, whilst seven sokemen together held 3 hides. (fn. 45) This land was granted by Alice de Clermont to the Knights Templars, and became known as Temple Hills Manor. (fn. 46) In 1199 they received a charter granting them assart of 200 acres of land in Sharnbrook, (fn. 47) and a charter of free warren was given them in 1253. (fn. 48) In 1276 the master claimed view of frankpledge twice yearly and 2 hides of land in Sharnbrook. (fn. 49) By virtue of the statute by which the Knights Templars' lands were transferred to the Knights Hospitallers, the prior of the latter order claimed Temple Hills Manor in 1331. (fn. 50) At this time it was leased to Sir John de Wolaston and Margaret his wife for the term of their lives. (fn. 51)
A fourth estate in Sharnbrook, known in the 16th century as LANGTONS MANOR, was held of the barony of Bedford. It may therefore have originated in the 1½ virgates of land which Osbern de Broilg held of Hugh de Beauchamp in 1086. (fn. 55) But it is impossible to state this definitely, for as will be seen from the number of tenants (the Druels, Parentines and Patishalls) who owed allegiance to this honour, the overlordship must have been considerably extended between the Survey and the early 13th century. It seems unlikely that Langtons was ever a manor in the technical sense, for, as mentioned above, it is not so called until very late.
The family of Osbern de Broilg remained in Sharnbrook for more than a century after the Survey, for in 1200 Robert de Broy acknowledged the right of Henry of Sharnbrook to a mill in Sharnbrook. (fn. 56) A gap then occurs in the descent, and the property reappears in 1324 as belonging to Robert de Utteford. (fn. 57) The estate, here alienated to John de Langton, is extended at one messuage, 80 acres of land, 2 acres of meadow and 6 acres of wood. (fn. 58) The Langtons, though they gave their name to the manor, do not appear to have held it long. Thomas de Langton held by feudal service in this parish in 1346, and in 1428 Thomas Bromflete held his fee. (fn. 59) Henry Bromflete Lord Vesey, son of Sir Thomas Bromflete, died in 1469 seised of land in Sharnbrook. (fn. 60) He left a daughter Margaret wife of Launcelot Threlkeld, who died in 1493. She alienated Langtons Manor, which appears in 1495 as a capital messuage, and 51s. 1d. rent, of which George Ingleton died seised, though how he acquired it has not been traced. (fn. 61) He left a son Robert, aged nine, who died in 1505. (fn. 62) His daughter Joan, nine months old at her father's death, subsequently married George Tyrrell, and dying in 1557 left a son George Tyrrell, aged twenty-eight, (fn. 63) and he in 1563 sold to Thomas Cobbe, the manors of Langtons (here so called for the first time) and Parentines for £240. (fn. 64) In 1573 Thomas Cobbe, whose family appear to have been settled in Sharnbrook for three generations, (fn. 65) died seised of Langtons Manor, which then passed to his son John. (fn. 66) He died in 1575 without attaining his majority, when Langtons and Parentines Manor passed to his brother Thomas, (fn. 67) then aged sixteen. He was followed by his son Paul Cobbe, (fn. 68) whose son Thomas Cobbe suffered a recovery of the manors in 1634. (fn. 69) He had several sons, of whom Paul held the manors in 1669. (fn. 70) William Cobbe was the only son to leave issue, a son William, who died in 1687 at the age of sixteen, when the manors passed to George Cobbe, a distant relation. (fn. 71) He conveyed the manor in trust in 1694, (fn. 72) but at his death without issue a partition of his property was made in 1700 between his sisters and co-heirs. (fn. 73) They were Elizabeth wife of James Watkins, Anne wife of John Aspin and Rebecca wife of Joseph Godfrey, and they appear to have held the manor conjointly until 1726. (fn. 74) It passed next to the Bullock family, and was held by John Bullock in 1766, (fn. 75) of whom it was purchased by Mr. Gibbard, (fn. 76) whose descendant Mr. L. Stileman Gibbard owns Langtons and Parentines Manors at the present day.
A small estate in Sharnbrook, later called PARENTINES MANOR, was also attached to the barony of Bedford, whose overlordship is acknowledged in 1278, 1302, 1346 and 1428. (fn. 77) The first mention that has been found of the Parentine family in Sharnbrook is amongst the benefactions to Newnham Priory, where Robert de Parentine's name occurs as having bestowed 13 selions of land in Haselhoo in this parish. (fn. 78) This would probably de early in the 13th century. In 1278 Ellen de Parentine held 1 hide in Sharnbrook, (fn. 79) and her name reappears in feudal aids for 1302 and 1316. (fn. 80) No further trace has been found of the Parentines, but by 1346 their fee appears to have become assimilated with Langtons Manor (q.v.). Parentines Manor, though following the same descent, did not lose its separate identity, which is mentioned as late as 1766. (fn. 81)
A sixth estate in Sharnbrook, known at the present day as OUSE MANOR, had its origin in the various grants made by the Child family to Newnham Priory about the 13th century. Robert son of William le Child gave the priory the waters of the Ouse inclosed from Sitegate to Besebrigg, (fn. 82) whilst John his son bestowed at various times 7, 5 and 20 selions of land. (fn. 83) In 1278 the prior was declared to have a free fishery from the mill of Pinch to the mill of Stoke and from Sculteleye to the head of the pond of Pinch. (fn. 84) This property followed the same descent as Tofte Manor, and it is not until 1614 that it becomes differentiated. It then appears in an inquisition as '5 messuages 246 acres of land and a piece of water called le Owes in Sharnebrook.' (fn. 85) It appears later in the Cobbe family, (fn. 86) though no record of the transfer has been found, and in 1700, on the partition of George Cobbe's estate amongst his sisters, it passed to Rebecca wife of Joseph Godfrey, who was holding it in 1721. (fn. 87) It was subsequently sold by Rebecca to Charles Dymoke William, (fn. 88) from whom it passed by sale to the father of Charles Tanqueray, who sold it before 1809 to Mr. Thomas Gell. (fn. 89) He left it by will to his nephew William Islip, whose mortgagees sold it in or about 1880 to the present owner, Mr. William Whitworth. (fn. 90)
Hugh the Fleming, brother of Walter, owned half a hide and one quarter of a virgate in Sharnbrook in 1086, (fn. 91) which is afterwards found attached to the barony of Wahull from the 13th to the 15th century. (fn. 92) Robert was the tenant of Hugh at Domesday, and this land passed later to a family called alternatively Rivers or Water. Richard Rivers held half a fee in Sharnbrook at the time of the Testa. (fn. 93) In 1278 the same Richard held half a hide, (fn. 94) which in 1302 had passed to Henry atte Water. (fn. 95) Alice atte Water was holding in 1346, (fn. 96) after which only one further mention has been found of this fraction of a knight's fee, when in 1428 William Herdewyk was declared to hold the quarter fee which Alice atte Water formerly held. (fn. 97)
Mention has been found of a seventh SHARNBROOK MANOR. Higham Ferrers College, which was founded by Henry Chicheley in 1422, (fn. 98) owned lands in Sharnbrook valued at £4 at the Dissolution. (fn. 99) In 1542 this land, here called Sharnbrook Manor, was granted to Robert Dacres, (fn. 100) who in the following year received a licence to alienate the property to George Boteler, (fn. 101) which became absorbed in Lorings (q.v.) and the other manors possessed by him in Sharnbrook.
In 1086 Osbern the Fisherman held half a hide in Sharnbrook, the Countess Judith held 3 virgates, and Almar, a burgess of Bedford, held half a virgate, of which estates no further mention has been found. (fn. 102)
Colworth, which is described as a hamlet attached to Sharnbrook as early as 1278, included a property held of the barony of Beauchamp of Bedford, known as COLWORTH MANOR. In 1278 it was held of William Fruel as of this barony in right of his wife, (fn. 103) and in 1417 was held of Baldwin Pigot, who had acquired rights of overlordship by descent. (fn. 104) The earliest tenants found of this manor were the Druels. John Druel held half a hide in Colworth in 1278–9, with free tenants, and 2 virgates in villeinage. (fn. 105) In 1304, together with Amice his wife, he obtained recognition of his right to the manor from William Warren. (fn. 106) In 1331 John son of John Druel acknowledged the right of his brother Giles to Colworth Manor. (fn. 107) No further trace has been found of the Druels, but within the next generation Colworth passed to the Greene family. Sir Thomas Greene died in 1391–2 holding by knight service in this hamlet, (fn. 108) leaving sons Thomas and Henry. (fn. 109) This manor appears to have passed to Henry Greene, the younger son, for c. 1415 John son of Henry Greene remitted to his brother Ralph all his claims on this property. (fn. 110) A year later Sir Ralph Greene, kt., and his wife Katharine are found placing this manor in the hands of William Islip and other trustees. (fn. 111) Ralph Greene died in 1417, when his brother John was declared to be his heir. (fn. 112)
This manor subsequently became Crown property, probably escheating during the Wars of the Roses. In 1484 Richard III granted it to Thomas Lynom and heirs male of his body 'for his good services against the rebels.' (fn. 113) Either the failure of issue of Thomas Lynom, or what is more probable, a forfeiture of the grant on the accession of Henry VII, brought this manor again into the possession of the Crown, and in 1546–7 Sir Edward Montague, kt., obtained a grant of it. (fn. 114) He died seised of this manor ten years later. (fn. 115) His son Edward, who was also knighted, died in 1602, leaving four sons, of whom Edward, the eldest, inherited the manor. (fn. 116) A gap here occurs in the history of this estate, which, according to Lysons, was purchased about the year 1700 by Mark Antonie, (fn. 117) who, dying in 1720, was succeeded by John Antonie, by whom the greater part of the present Colworth House was built. (fn. 118) He died in 1760, when this estate passed to his brother Richard Antonie, (fn. 119) who on his death without issue in 1771 left it by will to William Lee, who assumed Antonie as an additional name. (fn. 120) John Fiott, his nephew, succeeded William Lee-Antonie in 1815, assuming the name of Lee under his uncle's will. He was a great collector of pictures and other works of art, both ancient and modern, with which he enriched Colworth House. (fn. 121) He died in 1866, but previous to that date, in 1854, Colworth was sold to Hollingworth Magniac, who made it his residence until his death in 1867, when it passed by will to his eldest son Charles Magniac. (fn. 122) It subsequently became the property of the late William Clarence Watson, who modernized the house, which is now the residence of Mr. Albert Edward Bowen.
In 1340 the Prior of Caldwell owned 4s. rent in Sharnbrook. (fn. 123) At the Dissolution the temporalities of the priory here amounted to 5s., (fn. 124) and in 1540 were granted, together with Tofte Manor (q.v.), to George Boteler, and henceforward became absorbed in the larger estate. (fn. 125)
The church of ST. PETER consists of a chancel 32 ft. by 18 ft., north chapel of the same size, nave 46 ft. by 19 ft., north and south aisles 9 ft. 3 in. wide, west tower 17 ft. 5 in. by 12 ft. 4 in., and north and south porches.
The nave arcades show that in the 13th century the nave was 10 ft. shorter than at present, with north and south aisles; the chancel was about the same size as now. In the 14th century a large north-east chapel was added to the chancel, and a west tower built 10 ft. from the west end of the existing nave, or perhaps begun to the west of an existing tower. When the tower was completed the north and south walls of the nave were lengthened westwards to join it. A widening of the north aisle seems to have been intended, judging by the arch at the west of the north-east chapel; but, as so often happens in work of this date, the intention was abandoned, probably on account of the Black Death. In the 15th century the north aisle was extended westward to the tower, and perhaps rebuilt; and the south aisle was treated in the same way, another arch being added to the nave arcades on each side; the 14th-century window at the south-west of the south aisle must have been then moved to its present place.
The east window of the chancel is of three cinquefoiled lights of 15th-century style, the heads and tracery being modern, while the details of the jambs look like 14th-century work; but the chancel has a 15th-century plinth on its east and south walls, and appears to have been rebuilt early in that century. On the north side is a 14th-century arch of two orders with clustered responds opening to the chapel; on the south side are two windows—one of three lights and 15th-century tracery; the other, at the west, of two lights, with jambs of the same earlylooking section, but obviously of the same date as the 15th-century tracery. In its inner jambs are 14thcentury shafts with moulded capitals. A small south doorway between them with continuous moulded jambs and a moulded label also looks like 14thcentury work. The chancel arch has two hollowchamfered orders and responds with a semicircular shaft with a moulded capital to the inner order. The chancel roof is of low pitch, and is divided into three bays by plainly moulded tie-beams, perhaps of 17th-century date.
Plan of Sharnbrook Church
In the north chapel is a 15th-century east window of three lights in 14th-century jambs with internal shafts, capitals and bases; the head of the window is now four-centred, in order to suit the flat-pitched roof, but the older pointed head can still be seen on the inside. On the north side is a 14th-century square-headed window of three trefoiled lights with net tracery and moulded jambs, and further west is a plain early 17th-century doorway with a squareheaded window close to it on the west, to light a recess in the west wall, which has a trefoiled head and a stone seat. In the south wall is a range of three stepped 14th-century sedilia and a piscina, with trefoiled heads; they are separated by shafts with moulded capitals and bases. In the west wall, partly overlapped by the north wall of the aisle, is an arch moulded like that into the chancel, but resting on half-octagonal responds with plainly moulded capitals and bases. The roof is flat, and probably of the same date as that of the chancel. The nave arcades are of four bays, the first three on both sides being of the 13th century and the western bays of the 15th. The earlier work has arches of two hollow-chamfered orders with a label, the outer order having stops; the stop near the chancel arch on the south side is carved as a human head. The responds are semicircular, and of the columns the eastern pair are octagonal and the others circular; all have moulded capitals, that of the first column of the north arcade having a cable mould on one member. The stop to the label over this capital is a bishop with hand raised in blessing; and on the opposite side of the nave there is a bishop's head. The west arch in both arcades is of three chamfered orders with clustered responds and moulded capitals and bases. The nave roof is divided into four bays by 15th-century moulded principals, having carved and gilded bosses in the middle. The north aisle has three 15th-century windows, each of three cinquefoiled lights, but the cusping of the middle window has disappeared. The north doorway is of the date of the arcade and has a moulded arch and label, and in each jamb is a detached shaft with a moulded capital and base; it is probably in situ. Over it is a small 15th-century porch. The south aisle has an east window and two south windows like those in the north aisle, and at the south-west a three-light 14th-century window with net tracery. The south doorway, of the 15th century, is in two hollow-chamfered continuous orders and opens into a porch of the same date, which formerly had a roof of lower pitch than at present.
The tower, which opens to the nave by a massive pointed arch of three orders with moulded capitals and bases, has a large three-light west window with flowing tracery, and a stair at the south-west. It is finished with a parapet of pierced quatrefoils, and has the stump of a large pinnacle at each corner; above this rises a tall stone spire with three tiers of spire lights. The belfry is lighted on each side by a simple window of two lights. To the south of and above the west window of the ground stage are ogee-headed cinquefoiled niches, and below it is a sharp pointed four-centered doorway in two chamfered orders.
In the south aisle is a well-designed 15th-century octagonal font with quatrefoiled panelling, resting on an octagonal base with buttresses at each angle. The pewing is modern, but there is a good early 16th-century screen, with a crocketed ogee head to the middle opening, and on each side five traceried lights. The lower panels are solid, being on the south re-used 17th-century woodwork, but original on the north, with three holes cut through them in ancient times to give a view of the altar to anyone kneeling just outside the chancel. At the east end of the chancel is a wooden tablet to Thomas Cobbe, 1662, with his arms Gules a cheveron wavy between three cobfish argent, and a chief argent with three ravens therein. Over the arch on the north side of the chancel is a similar wooden tablet to William Cobbe, his son, 1675, with his arms impaling Azure a cheveron between three leopards or. In the north chapel is a large modern monument of 18th-century design to the Magniac family of Colworth. In the floor is a slab with the name of William Cobbe, 1687. In the north chapel is a fine Jacobean mural monument, with the Boteler arms, three covered cups, inclosing a brass panel, set up by Sir William Boteler, with a pedigree of his family on it. There are two 17th-century chests at the west end of the north aisle.
There are six bells: the treble, cast by John Warner, 1887; the second, by Henry Bagley, 1683; the third and fourth, by Matthew Bagley, of the same date; the fifth, by Newcombe of Leicester, 161–, the last figure being omitted; and the tenor, by Henry Bagley, 1699.
The communion plate is modern. A silver chalice and paten were given by Miss Bowen in 1910.
The earliest book of registers contains baptisms and burials 1596 to 1653 and marriages 1596 to 1676; the second contains baptisms and burials 1653 to 1765 and marriages 1676 to 1754; the third contains births and deaths 1766 to 1812; and the fourth marriages 1754 to 1812.
The church of Sharnbrook was granted to the abbey of St. Mary de Pré, Leicester, by Baldwin Triket in the reign of King John. (fn. 126) Its value in 1291 was £13 6s. 8d., (fn. 127) and at the Dissolution the vicarage was worth £7 9s. 8d. and the rectory £11. (fn. 128) The advowson then passed to the Crown, with whom it remained (fn. 129) until 1877, in which year Charles Magniac presented, having purchased the right under Lord Westmorland's Act of 1863. (fn. 130) The present patron is Mr. Vernon Magniac.
Edward Earl of Lincoln and others received a grant of Sharnbrook Rectory in 1577–8, (fn. 131) which in 1586 had passed to Lord St. John of Bletsoe. (fn. 132) Oliver Lord St. John died seised of the rectory and tithes in 1618. (fn. 133) His son was still in possession in 1639, (fn. 134) but between this date and 1669 alienated the rectory to Paul Cobbe, who held it at that date. (fn. 135) In 1694 George Cobbe alienated to John Orlebar, (fn. 136) whose family retained the rectory as late as 1734. (fn. 137) By 1770 it had passed to William Gibbard, who died in 1800, when, with his other estates, it passed to John Gibbard his son, (fn. 138) who was impropriator at the time of the inclosure of the parish, when the great tithes were commuted for 239 acres of land. (fn. 139)
At the dissolution of the chantries Sharnbrook Church had an endowment of 30d., due from rents in the parish, to supply an obit and lamp. (fn. 140)
In 1827 William Curtis by will bequeathed £200 upon trust to be invested and income applied for the benefit of poor frequenting the meeting-house of Particular Baptists. The legacy with accumulations is represented by £265 17s. 11d. consols, with the official trustees, producing yearly £6 12s. 8d.


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What Is Timbre?
A. Leverkuhn
Timbre is a term for specific qualities of sound, usually applied to musical sounds. Many people refer to timbre as "sound quality," because differences in this category of sound can create more appealing results. This term for sound quality is generally applied to sounds that have a musical tone, rather than percussion or other less tonal sounds.
Many people confuse timbre with the frequency or pitch of musical tones. Although musicians and others often speak of specific musical notes as tones, the sound quality called tone sometimes applies to a specific point on a musical scale. Beginners can think of this as the common musical scale "do re mi fa…" or the conventional letter scale for music: A, B, C, D, E, F, G. These and other smaller incremental changes are referred to as tone or pitch changes.
Man playing a guitar
For many who are not heavily involved in music theory, timbre is a general term for how a particular instrument or voice sounds. Some people will analyze it by the treble and base components, or the high and low registers, of a particular instrument sound. Others will use advanced physics concepts to analyze timbre.
According to the physics of sound, some technical aspects of timbre include harmonic content, tremolo, or vibrato, and something called the attack-delay envelope. To understand the attack-delay envelope, it's helpful to see the musical sound represented by a wave pattern in a software application. Here, those who are looking at sound timbres for the first time can see how specific sounds generate wider waves at different points. For example, a cymbal sound may have a wider wave at the beginning or in the "attack" of the sound.
Harmonic content can also be represented in wave forms of sounds. In general, harmonics relate to a fundamental frequency for sounds. All of this involves highly technical evaluations of sounds, where less experienced listeners will usually judge the timbres of sounds by certain analog or basic factors, such as whether the music sounds "tinny" or "nasal" in a treble range, or "muddy" or "booming" in a bass range.
Musicians can evaluate timbre for many different reasons. They can do it to try to match the right instrument to a piece of music, where a brass horn or other instrument choice sounds better than others. They can also use timbre as part of an assessment of an individual instrument available for purchase, or for one that has developed a crack or other defect in need of repair. Talking about the timbres of sounds is often practical in a studio environment as well, as mutes and other devices and techniques can be use to modify the normal timbre of a particular instrument.
What Is a Tenor?
What Is Messa Di Voce?
What Is a Musical Tone?
What Is a Soprano Cornet?
What Is a Tenor Saxophone?
What is a Tone Decoder?
What is a Mezzo Soprano?


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Klotho, an anti-aging gene, acts as a tumor suppressor and inhibitor of IGF-1R signaling in diffuse large B cell lymphoma
Xiangxiang Zhou1,
Xiaosheng Fang1,
Yujie Jiang1,
Lingyun Geng1,
Xinyu Li1,
Ying Li1,
Kang Lu1,
Peipei Li1,
Xiao Lv1 &
Xin Wang1,2
Klotho, is a transmembrane protein, performs as a circulating hormone and upstream modulator of the insulin-like growth factor-1 receptor (IGF-1R), fibroblast growth factor (FGF), and Wnt signaling pathways. These pathways are involved in the development and progression of B cell lymphoma. We aimed to explore the expression pattern and functional mechanism of Klotho in diffuse large B cell lymphoma (DLBCL).
Immunohistochemistry (IHC) and western blotting were performed to detect the expression level of Klotho in DLBCL patients and cell lines. Tumor suppressive effect of Klotho was determined by both in vitro and in vivo studies. Signaling pathway activity was assessed by western blotting.
Remarkable lower expression levels of Klotho were observed in DLBCL patients and cell lines. Enforced expression of Klotho could significantly induce cell apoptosis and inhibit tumor growth in DLBCL. Upregulation of Klotho resulted in declined activation of IGF-1R signaling, accompanied with decreased phosphorylation of its downstream targets, including AKT and ERK1/2. Moreover, xenograft model treated with either Klotho overexpression vector or recombinant human Klotho administration presented restrained tumor growth and lower Ki67 staining.
Our findings establish that Klotho performs as a tumor suppressor and modulator of IGF-1R signaling in DLBCL. Targeting Klotho may provide novel strategies for future therapeutic intervention.
Diffuse large B cell lymphoma (DLBCL) is the most common form of non-Hodgkin lymphoma (NHL), accounts for nearly 40% of all newly diagnosed cases [1]. This disease presents as an aggressive process and exhibits high heterogeneity in gene expression and clinical outcomes [2, 3]. Although majority of DLBCL patients could be cured by anthracycline-based chemotherapies combined with rituximab, one third of them presented refractory or relapsed process [4, 5]. Therefore, more effective treatment strategies based on novel therapeutic targets and molecular oncogenic pathways are still needed.
Klotho is an anti-aging gene originally identified in 1997 [6]. Kuro-o et al. [6, 7] found that Klotho-deficient mice developed multiple premature aging syndromes, whereas overexpression of Klotho extended the lifespan of mice. The Klotho gene is located in chromosome 13q12 in human with the size of 50 kb [6]. It encodes a single-pass transmembrane protein, which consists of an extracellular domain, a single transmembrane domain, and an intracellular domain. The intracellular domain is very short and has no clear functions. Membrane Klotho functions as an obligate co-receptor of fibroblast growth factor 23 (FGF23) to regulate phosphate homeostasis [8]. The extracellular domain (secreted Klotho) could be released into the serum and functions as a circulating hormone to regulate the activity of oxidative stress, multiple growth factor receptors, and ion channels [9, 10].
The tumor suppressive activity of Klotho was first identified in breast cancer in 2008 [11]. Recent investigations have implicated that Klotho is extensively downregulated in several solid tumors, including cervical cancer, pancreatic cancer, melanoma, and several digestive neoplasm [12]. In these malignancies, Klotho was elucidated to be a modulator of several signaling pathways, including the FGF signaling, insulin-like growth factor-1 receptor (IGF-1R), and Wnt pathways, which are also involved in the pathogenesis of hematological malignancies [10, 13–15]. However, the role of Klotho in hematological malignancies has not been reported.
A large number of aberrant receptor tyrosine kinases (RTKs) have been found in hematological malignancies [16, 17], but they are still indefinite in DLBCL. IGF-1R is a RTK primarily activated by its cognate ligands, IGF-1, and IGF-2. It plays a crucial role in the establishment and progression of tumors by regulating proliferation, self-renewal, apoptosis, and drug resistance of cancer cells [18–20]. Activation of IGF-1R following IGF-1 treatment results in phosphorylation of downstream signaling cascades, including PI3K/AKT and MAPK/ERK [19]. Blockade of PI3K/AKT signaling could restrain cell survival and function of lymphocytes [21, 22]. Activation of MAPK/ERK signaling promotes cell proliferation and metastasis of cancer cells [23]. Strategies to block IGF-1R pathway in solid malignancies are being tested in clinical trials [24], whereas the function of IGF-1R signaling in DLBCL has been less studied [25].
In this present study, we aimed to assess the expression level and functional mechanism of Klotho in DLBCL. We identified reduced expression of Klotho in DLBCL for the first time. Noted inhibition of cell growth and induction of apoptosis were observed in DLBCL with Klotho overexpression. Tumor growth was restrained by administration of Klotho protein in xenograft model of DLBCL. Our findings demonstrated that Klotho was a tumor suppressor and modulator of IGF-1R signaling in DLBCL, indicating that targeting Klotho may provide novel therapeutic strategy in DLBCL.
This study was approved by the Medical Ethical Committee of Shandong Provincial Hospital affiliated to Shandong University. The paraffin-embedded archived samples from 50 newly diagnosed DLBCL patients and 20 reactive hyperplasia patients were collected. Samples of patients with reactive hyperplasia were referred as control. Histological diagnoses were established according to the WHO classification [26]. Peripheral blood mononuclear cells (PBMCs) from the whole blood of healthy donors were isolated using Ficoll-Hypaque density gradient centrifugation (TBD science, Tianjin, China). Normal peripheral blood CD19+ B cells were purified from freshly isolated PBMCs using CD19+ magnetic microbeads kit (Miltenyi Biotec, Bergisch Gladbach, Germany). Cells were incubated with beads for 15 min at 4 °C while rotating. Purified CD19+ B cells were selected according to the manufacturer's protocols. The purity of isolated populations was assessed by FACS analysis, and cells with >90% purity were collected. All samples were obtained with informed consent in accordance with the Declaration of Helsinki.
Cell lines and reagents
Human DLBCL cell lines LY1 and LY8 were cultured in Iscove modified Dulbecco medium with 10% heat-inactivated fetal bovine serum. The medium contains 1% penicillin/streptomycin mixture and 2 mmol/l glutamine. CD19+ B cells and PBMCs obtained from three healthy donors were used as controls (N1, N2, and N3 cells). Recombinant human Klotho (rhKL) and recombinant human IGF-1 were obtained from PeproTech (Rocky Hill, NJ, USA), and adriamycin (ADR) was bought from Actavis (S.p.A, Italy).
Cell transfection
Lentivirus vectors either encoding Klotho or an empty lentiviral vector were from Genechem (Shanghai, China). Lentivirus transfection was carried out according to the manufacturers' instruction. Infection efficiencies were assessed by green fluorescent protein (GFP) through flow cytometry. The stably transfected cells were selected 48 h later with 5 μg/ml puromycin (Sigma-Aldrich, USA).
Quantitative real-time PCR
Total RNA was extracted using RNAiso Plus (TaKaRa, Dalian, China). Reverse transcription reaction was conducted with the reverse transcription reagents (TaKaRa, Dalian, China). Amplification reactions were performed with SyberGreen (TaKaRa, Dalian, China) in LightCycler 480II (Roche, Basel, Swizerland). Klotho-specific primers were as follows: forward, 5′-AGCAATCTGGTCTGAATAACACTGG; reverse, 5′-CATGTTTCAGCGTGAAAGTTCAAAG. Relative quantification was calculated using the △△CT method.
Immunohistochemistry (IHC) and hematoxylin-eosin staining
The 4-μm-thick formalin-fixed, paraffin-embedded tissue sections were deparaffinized and hydrate. Antigen retrieval was performed using 0.01 mol/l sodium citrate buffer (pH 6.0) under high pressure followed by a 1-h cool-down and rinses in phosphate buffer solution (PBS). Endogenous peroxidase was blocked with 3% hydrogen peroxide in methanol for 15 min, followed by incubation with normal serum to block non-specific binding. The slides were then incubated overnight at 4 °C with primary antibodies, anti-Klotho (1:150) or anti-Ki67 (1:100). After washing, the tissue sections were treated with the second antibody from SP reagent kit (Zhongshan Goldenbridge, Beijing, China) for 30 min at room temperature, followed by further treated with strept avidin-horseradish peroxidase complex (SABC) for 30 min at room temperature. After treated with diaminobenzidine (DAB) Kit (ZhongshanGoldenbridge, Beijing, China), the stained slides were counterstained with hematoxylin and mounted. Negative control was carried out with the primary antibody replaced by PBS. IHC staining was scored by the proportion of positive tumor cells. Five microscopic fields with the highest immunoreactivity at ×400 magnification were evaluated by two independent observers who were blinded to the patients' clinical data. Cases with at least 10% of tumor cells with Klotho staining were considered as positive. Fresh mice subcutaneous tumors were fixed in 4% paraformaldehyde (PFA) and embedded with paraffin for histological examinations. Sections with 4-μm thickness were cut and stained with hematoxylin-eosin (H&E).
Cells were lysed in radio-immunoprecipitation assay buffer (Shenergy Biocolor, Shanghai, China) together with 1× phosphatase inhibitor cocktail (PhosSTOP; Roche, Mannheim, Germany). The BCA assay (Shenergy Biocolor, Shanghai, China) was performed to detect protein concentration. Protein extracts (30 μg) were then electrophoresed on SDS-polyacrylamide gels and blotted from the gel onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). Membranes were incubated with the blocking solution (5% skim milk in Tris-buffered saline containing 0.05% Tween-20) for 1 h at room temperature and then immunoblotted with the indicated antibodies (1:1000 dilution) overnight at 4 °C. After which, the membranes were washed with TBS-T and then probed with the HRP-conjugated secondary antibodies (Zhongshan Goldenbridge, Beijing, China) and the electro-chemi-luminescence kit (Millipore, Billerica, MA, USA). Chemiluminescent signals were detected using the Amersham Imager 600 imaging system (General Electric, USA). ImageJ software (ImageJ, NIH) was used to quantify the protein bands normalized to control. Primary antibodies used were Klotho (Abcam, Cambridge, MA, USA), phospho-IGF-1R (Tyr1135/1136), IGF-1R, phospho-AKT(Ser473), total pan-AKT, Mcl-1, Caspase-3, diphosphorylated and total ERK1/2 (Cell Signaling Technologies, Beverly, MA, USA), β-actin, and GAPDH (Zhongshan Goldenbridge, Beijing, China). The experiments were performed in triplicate with GAPDH or β-actin (Zhongshan Goldenbridge, Beijing, China) as endogenous control.
Cell proliferation assay
Cell proliferation was assessed by performing triplicate assays with the Cell Counting Kit-8 (CCK-8) assay (Enogene, Nanjing, China). DLBCL cells with designed treatment were seeded in 96-well plates at a density of 5000 cells/well for 48 or 24~96 h later. Thereafter, the cells were incubated with 10 μl/well CCK-8 for 4 h according to the manufacturer's proposal. Cell proliferation was detected by light absorption at 450 nm by Multiskan GO Microplate Reader (Thermo Scientific, Rockford, IL, USA).
Flow cytometry analysis
Apoptosis of transfected DLBCL cells were detected by Annexin V-PE/7-aminoactinomycin (7AAD) (BD Biosciences, Bedford, MA, USA) assay according to the manufacturers' instructions. DLBCL cells with designed treatments were harvested and washed twice with ice-cold PBS and incubated in 1× binding buffer (containing 5 μl Annexin V-PE and 5 μl 7AAD). After incubation in the dark for 15 min, cells were subjected to the flow cytometry. At least 10,000 events per sample were acquired. Cells were discriminated into viable cells (AnnexinV-PE−/7AAD−), dead cells (Annexin V-PE−/7AAD+), apoptotic cells (Annexin V-PE+/7AAD−), and necrotic cells (Annexin V-PE+/7AAD+). The rates of apoptotic cells were acquired on a FACS-Navios Flow Cytometer (Beckman Coulter, CA, USA). Data were analyzed using FlowJo Version 7.6 software (Tree Star Inc., OR, USA).
Elisa assay
Serum soluble Klotho levels were measured using the Elisa kit (Immuno-Biological Laboratories, Gunma, Japan), with a lower limit of assay of 6.15 pg/ml.
In vivo xenograft study
All animal experiments were performed in accordance with the principles of the Institutional Animal Care. Severe combined immunodeficiency (SCID) Beige female mice of 5-week old were bought (Weitong Lihua Laboratory Animal Center, Beijing, China) and maintained in a pathogen-free environment under controlled condition of light and humidity. 1 × 107 LY1 cells (untransfected, stably Klotho-overexpressing vector tranfected, or empty control vector transfected, respectively), mixed with 100 μl Matrigel (BD Biosciences, Bedford, MA, USA), were subcutaneously injected into the right inferior limb of mice. Tumor size was measured by the digital caliper. For the experiment with rhKL, SCID Beige mice were injected subcutaneously with 1 × 107 LY1 cells into the left inferior limbs. Mice were treated with daily intraperitoneal injections of rhKL (7.5 μg/kg) or PBS control (six mice per group) for 2 weeks. The volume of tumor was estimated using the equation V = (a × b 2) × 0.5236, where a is the largest dimension and b is the perpendicular diameter.
All statistical analyses were performed by using the statistic software SPSS17.0 (SPSS Inc., Chicago, IL, USA) for Windows. In vitro experimental results were presented as mean ± SD of data obtained from three separate experiments. Overall survival time was measured from the date of diagnosis to the date of death or the last follow-up. Kaplan-Meier analysis was performed to estimate the survival functions. A log-rank test was used to assess survival differences. Chi-square test was used to analyze the clinical characteristics of patients. One-way analysis of variance (ANOVA) or t tests were used to assess the differences between groups. p < 0.05 was considered to be statistically significant.
Klotho was downregulated in DLBCL and related to tumor progression
Compared with reactive hyperplasia, expression level of Klotho was significantly lower in DLBCL tissues (Fig. 1a). Klotho positive rate was 14% (7 of 50) in DLBCL whereas 80% (16 of 20) in reactive hyperplasia. To evaluate the clinical significance of Klotho in DLBCL, clinical and pathological characteristics of DLBCL patients were analyzed. The expression level of Klotho was associated with the Ann Arbor stage of DLBCL patients. Patients with stage III or IV presented markedly lower Klotho level than those with stage I or II (p = 0.002, Table 1). In addition, Kaplan-Meier survival analysis were performed based on the expression levels of Klotho (n = 50). The median survival of patients in Klotho-positive group was 48.46 months, significantly longer than those in Klotho-negative group (29.27 months, p = 0.045, Fig. 1b). We next confirmed the expression of Klotho in DLBCL cell lines. Klotho mRNA expression was detected in DLBCL cell lines and normal CD19+ B cells. Significantly decreased levels of Klotho mRNA expression were observed in DLBCL cell lines (LY1 0.024 ± 0.037, LY8 0.002 ± 0001, Fig. 1c). Reduced expression levels of Klotho protein were also noted in DLBCL cell lines compared to human PBMCs (Fig. 1d).
Klotho was downregulated in DLBCL and related to tumor progression. a Compared with reactive hyperplasia, expression level of Klotho was significantly decreased in DLBCL tissues. Original magnification, ×100 (upper panel) and ×400 (lower panel). b Patients with Klotho negative expression showed shorter survival than those with Klotho positive expression. c As detected by real-time quantitative PCR, lower levels of Klotho mRNA expression were observed in DLBCL cell lines (LY1, LY8) than in CD19+ B cells (N1, N2, N3) (mean ± SD, n = 3, *p < 0.05). d Protein expression levels of Klotho were detected in DLBCL cells and normal PBMCs. The ratios of relative protein expression level of targets are indicated below the western blot
Table 1 Correlation between Klotho expression and clinical characteristics of DLBCL patients
Klotho inhibited growth of DLBCL
To further establish the biological function of Klotho, human DLBCL cells were stably transfected with either negative control lentivirus vectors (LV-Con) or Klotho-overexpression lentivirus vectors (LV-KL). The upregulation of Klotho levels were confirmed by quantitative PCR and western blot (Fig. 2a, b). Cell proliferation rates were evaluated by CCK-8 assay. Significant reduction of cell proliferation was observed in LY1 and LY8 cells transfected with LV-KL, compared with those transfected with empty vector (Fig. 2c). Furthermore, we investigated the anti-tumor effect of Klotho on DLBCL xenografts in vivo. LY1 cells, stably expressing Klotho, or the vector control cells were injected subcutaneously into SCID Beige mice, respectively. Consistent with the in vitro results, mice treated with Klotho overexpression cells revealed remarkable reduction in tumor volume compared to that transfected with empty vector (Fig. 2d). Higher expression level of Klotho was identified in mice treated with LV-KL-transfected LY1 cells (Fig. 2e). In addition, we estimated the expression level of proliferative marker Ki67 [27, 28] in xenograft mice by IHC. Lower level of Ki67 positive rate were observed in LV-KL group (Fig. 2e).
Klotho suppressed DLBCL growth. a, b Relative expression levels of Klotho were confirmed by quantitative PCR (mean ± SD, n = 3, **p < 0.01) and western blot in stably transfected LY1 and LY8 cells compared to empty vectors. The ratios of relative protein expression level of targets are indicated below the bands. c DLBCL cells transfected with LV-KL presented significantly lower level of cell proliferation than those transfected with empty vector (mean ± SD, n = 3, **p < 0.01). d SCID mice with Klotho overexpression revealed significantly lower tumor volume than those with empty vector (n = 6 per group, **p < 0.01). e H&E staining and IHC staining of Ki67 and Klotho were performed in mice tumors. Original magnification, ×400
Klotho promoted apoptosis of DLBCL cells
To investigate whether Klotho could promote the apoptosis of DLBCL cells, Annexin-V-based apoptotic assays were performed. Flow cytometry analysis indicated that Klotho overexpression resulted in enforced apoptosis rates in both LY1 (6.56 ± 0.71% in LV-Con vs. 16.41 ± 2.16% in LV-KL group, p = 0.002) and LY8 cells (7.23 ± 0.65% in LV-Con vs. 16.03 ± 1.85% in LV-KL group, p = 0.001, Fig. 3a, b). In addition, western blot analysis confirmed the pro-apoptotic effects of Klotho. Dramatically reduction of anti-apoptotic protein Mcl-1 and increased cleaved forms of Caspase-3 were observed in DLBCL cell lines (Fig. 3c). Altogether, these results indicate that upregulation of Klotho could promote the apoptosis of DLBCL cells.
Klotho promoted apoptosis of DLBCL. a, b Enforced expression of Klotho resulted in increased apoptosis rates in LY1 and LY8 cells assessed by flow cytometric analysis with Annexin V-PE/7AAD staining (mean ± SD, n = 3, **p < 0.01). c Declined expression levels of Mcl-1 and increased levels of activated Caspase-3 were observed in LV-KL-treated DLBCL cells. The ratios of relative protein expression level of targets are indicated below the bands
Klotho modulated the activation of IGF-1R signaling in DLBCL
Having shown that Klotho could impair cell proliferation and induce apoptosis in DLBCL, we next investigated the molecular mechanisms responsible for the function of Klotho. The IGF-1R pathway plays a vital role in the development of hematological malignancies [14, 25, 29]. CCK-8 assay was conducted to assess the effect of Klotho on IGF-1-induced cell proliferation. DLBCL cells transfected with either LV-KL or LV-Con were treated with IGF-1 or vehicle control in 0.5% FBS culture medium for 24–96 h. In the groups untreated with IGF-1, LV-KL transfection resulted in declined proliferation of LY1 and LY8 cells compared to empty-vector group. In the IGF-1-treated groups, we observed that cell proliferation was less restored by IGF-1 in cells transfected with LV-KL compared to that transfected with LV-Con. Following addition of IGF-1, cell proliferation of LV-Con-treated cells increased by up to 60%, whereas the only up to 40% enhancement of cell proliferation was found in LV-KL transfected cells (Fig. 4a).
Klotho modulated activation of IGF-1R pathway in DLBCL. a IGF-1-induced DLBCL cell (LY1 and LY8) proliferation is inhibited by Klotho overexpression. LY1 and LY8 cells were transfected with LV-KL or LV-Con, starved for 48 h and treated by IGF-1 (50 ng/ml) and analyzed by CCK-8 assay (mean ± SD, n = 3, *p < 0.05, **p < 0.01). b LY1 with stable transfection of LV-KL or LV-Con were serum starved for 48 h and treated with IGF-1 (50 ng/ml) for the indicated times or IGF-1 (30 min) for the indicated doses. After treatment, cells were harvested and analyzed by western blot. c LY1 and LY8 cells transfected with LV-KL or LV-Con, serum starved for 48 h, and treated with IGF-1 (50 ng/ml, 30 min). Western blot was conducted to assess the phosphorylated (p) and total (t) protein levels of IGF-1R, AKT, and ERK1/2. The ratios of relative protein expression level of targets are indicated below the bands. d Decreased activation of IGF1-R signaling was observed in LV-KL-treated mice. The ratios of relative protein expression level of targets are indicated below the bands. e Schematic description of Klotho mediated IGF1R signaling
As the IGF-1R signaling could be activated by IGF-1, we explored the optimal activation dose and time of IGF-1 in DLBCL cell lines. LY1 cells were transfected with either LV-KL or LV-Con, serum starved for 48 h and treated with IGF-1 for the indicated times and doses. Western blot was carried out to evaluate the phosphorylation level of the IGF-1R. As shown in Fig. 4b, the maximum activation occurred at IGF-1 concentration of 50 ng/ml and at the time of 30 min after treatment. Then, we studied the ability of Klotho to modulate activation of IGF-1R signaling in DLBCL cells. Cells transfected with either LV-KL or LV-Con were serum starved for 48 h, treated with IGF-1 (50 ng/ml for 30 min) or vehicle control. Following treatment, cells were harvested and immunoblotting was conducted. Decreased phosphorylation level of IGF-1R and its downstream targets, including AKT and ERK1/2, were observed in cells transfected with LV-KL (Fig. 4c). Furthermore, we evaluated the modulation of Klotho on IGF-1R signaling in DLBCL xenograft model. Enhanced expression of Klotho in LV-KL-treated mice was confirmed by immunoblotting (Fig. 5d). Decreased phosphorylation of IGF1-R as well as its downstream targets were observed in mice treated with LV-KL compared to the control group (Fig. 4d). These results demonstrated that Klotho may act as a modulator of IGF-1R signaling contributed to the tumorigenesis of DLBCL (Fig. 4e).
rhKL acted as an active form in vitro and vivo. a, b LY1 and LY8 cells were treated with rhKL, ADR, their combination, or a vehicle control. CCK-8 assay was conducted after 48 h (mean ± SD, n = 3, *p < 0.05 for comparison between treated cells and control, **p < 0.05 between Klotho and ADR combination versus ADR alone). c LY1 cells were injected subcutaneously into the left inferior legs of SCID Beige mice. The mice were treated with daily intraperitoneal injections of rhKL (7.5 μg/kg) or vehicle control (PBS) for 2 weeks. Tumor volumes were measured every 2 days (n = 6 per group, *p < 0.05). d H&E staining and IHC staining with Ki67 were performed. Original magnification, ×400. e Lower serum Klotho levels were detected by Elisa in DLBCL patients than the control subjects
rhKL acted as an active form both in vitro and in vivo
We further investigated the activity of secreted Klotho protein on DLBCL. To explore the effect of secreted Klotho on proliferation of DLBCL cells, LY1 and LY8 cells were seeded and treated with rhKL. The rhKL was found to be active and reduced the proliferative ability of DLBCL cells by 30~40% at the concentration of 2 μg/ml (Fig. 5a, b). ADR is one of the most commonly used drugs in the chemotherapeutic strategy of DLBCL. We detected the function of Klotho on cell responses to ADR. LY1 and LY8 cells were treated for 48 h with rhKL, ADR, or their combination, and cell proliferation was then evaluated. Combination of rhKL (2 μg/ml) increased the sensitivity of DLBCL cells to ADR (Fig. 5a, b).
The in vivo activity of rhKL in DLBCL xenograft model was also detected. LY1 cells were injected subcutaneously into the SCID Beige mice to build a DLBCL xenograft model. After 15 days, the mice were treated with daily intraperitoneal injection of either rhKL (7.5 μg/kg, n = 6) or a control vehicle (n = 6) for 2 weeks. Significant decreased tumor volumes were observed in the mice treated with rhKL compared with those treated with vehicle control (Fig. 5c). Moreover, reductive expression level of proliferative marker Ki67 was found in rhKL-treated mice (Fig. 5d).
In addition, we employed ELISA assays to test the soluble Klotho levels in the serum of 30 initially diagnosed DLBCL patients and 15 healthy volunteers. Lower serum Klotho levels were noted in DLBCL patients (628.54 ± 219.39 pg/ml) than the control subjects (818.87 ± 241.51 pg/ml, p = 0.045, Fig. 5e).
In this study, our observations identified for the first time that Klotho, an anti-aging gene, as a potential tumor suppressor in DLBCL tumorigenesis. Klotho was downregulated in human DLBCL and inhibited the growth of DLBCL both in vitro and in vivo. Upregulation of Klotho resulted in declined activation of IGF-1R signaling pathway in DLBCL.
We identified remarkable reduced expression of Klotho in DLBCL tissues and cell lines, whereas higher expression in reactive hyperplasia and CD19+ B cells from normal donors. Decreased level of Klotho was associated with advanced stage and more aggressive disease process in DLBCL. Tumor suppressive activity of Klotho has been reported in several human solid malignancies, but never in hematological cancers [30–33]. Recent investigations elucidated that epigenetic mechanisms, including promoter methylation and histone deacetylation, contributed to the reduced Klotho expression in human breast cancer, cervical cancer, and hepatocellular carcinomas [34–36]. The similar mechanism may be involved in DLBCL. At present, the diagnosis of DLBCL is mainly based on the biopsy histopathology and IHC [37]. With the deepening of additional studies, lower expression of Klotho may serve as a potential marker for the pathological diagnosis of DLBCL.
DLBCL is a high-grade heterogeneous disorder defined by heterogeneity in clinical and biological characteristics [1, 38, 39]. Personalized prognostic stratification and targeted therapeutic strategies are urgently needed to improve the outcomes of DLBCL patient [40]. Ki67 as a proliferative marker performed as a poor prognostic marker in DLBCL [27]. In this study, we discovered that DLBCL xenograft mice with Klotho overexpression exhibited significantly lower Ki67 staining positive rate than that without Klotho upregulation.
As Klotho exists in both membrane-bound form and secreted form, the secreted Klotho could be shed and released into the circulation [7]. Recent investigation elucidated the low serum level of Klotho in renal cell carcinoma [41]. Low serum Klotho level was an independent adverse prognostic factor for cancer-specific and progression-free survival in RCC [41]. However, the level of serum soluble Klotho was unchanged in multiple myeloma [42]. Thus, the diagnostic role of secreted Klotho in human cancer remains controversial. In this study, we identified the decreased level of serum Klotho in DLBCL patients. The correlation of serum Klotho with disease diagnosis and progression still needs further exploration. Larger number of the included patients will better confirm the role of serum Klotho in DLBCL.
Importantly, our study illuminated that Klotho effectively inhibited the growth of DLBCL cells. Overexpression of Klotho significantly inhibited cell proliferation and induced cell apoptosis in DLBCL. We also discovered that efficacy of ADR could be enhanced by combination with rhKL in DLBCL cells. This finding suggested the potential of Klotho in therapeutic intervention of DLBCL. Our in vivo investigations demonstrated that upregulation of Klotho, either by LV-KL transfection or rhKL administration, congruously led to inhibitory effect in the tumor growth of xenograft model of DLBCL. This will pave the way for rational design of Klotho-based molecular products in DLBCL. Nevertheless, pharmacokinetic investigations are still required to explore the optimal dose and adverse reaction of rhKL in the use of either single agent treatment or drug combination.
Significantly, we also found that Klotho could inhibit the activation of IGF-1R signaling in DLBCL. Induction of IGF-1R signaling was involved in the pathogenesis of hematological malignancies [13]. Recently, Stromberg et al. reported the involvement of IGF-1R and the antitumor effects of specific IGF-1R inhibitors in DLBCL tumorigenesis [25]. Structure-function analysis of Klotho indicated that Klotho could interact with the IGF-1R [43]. Klotho-induced inhibition of IGF-1R signaling may act as a novel mechanism involved in the development of DLBCL. Apart from IGF-1R signaling, many other pathways have been confirmed to be modulated by Klotho in tumorigenesis. The most studied is the FGFR-Klotho axis, and Klotho could act as cofactor of endocrine FGFs to bind and induce the activation of FGFRs in breast, pancreatic, and prostate cancers [11, 30, 44]. Deregulation of Wnt signaling pathway plays a critical role in the pathogenesis of cancers [45–47]. It was proved that Klotho participated in tumorigenesis partly through restraining the Wnt signaling pathway [48]. The modulation of Klotho on these pathways in DLBCL is poorly understood. In spite of the important discoveries revealed by these studies, there are also limitations on the detailed mechanisms and the cross-talks of them involved in Klotho deregulation.
Taken together, our findings identified that Klotho performs as tumor suppressor and modulator of IGF-1R signaling in the DLBCL. Overexpression of Klotho may be a predictive marker for favorable outcome in DLBCL. Klotho reinforces the response of DLBCL cells to chemotherapeutic drug. Being an endogenous circulating hormone, the secreted Klotho could function as an active form and inhibit the tumor growth effectively both in vitro and in vivo. This study illuminates Klotho as a potential target for future therapeutic strategies.
CCK-8:
DAB:
Diaminobenzidine
DLBCL:
Diffuse large B cell lymphoma
H&E:
Hematoxylin-eosin
IGF-1R:
Insulin-like growth factor-1 receptor
IHC:
International prognostic index
LDH:
Lactate dehydrogenase
LV-Con:
Negative control lentivirus vectors
LV-KL:
Klotho-overexpression lentivirus vectors
NHL:
PBMCs:
Peripheral blood mononuclear cells
PBS:
Phosphate buffer solution
PFA:
rhKL:
Recombinant human Klotho
RTKs:
SABC:
Strept avidin-horseradish peroxidase complex
SCID:
Severe combined immunodeficiency
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This study was supported by National Natural Science Foundation (No. 81473486 and No. 81270598), National Public Health Grand Research Foundation (No. 201202017), Natural Science Foundations of Shandong Province (No. ZR2012HZ003 and No. 2009ZRB14176), Technology Development Projects of Shandong Province (No. 2014GSF118021, No. 2010GSF10250, and No. 2008GG2NS02018), Program of Shandong Medical Leading Talent, and Taishan Scholar Foundation of Shandong Province.
All data generated and analyzed during this study are included in this published article.
XXZ and XW conceived and designed the experiments. XXZ, XSF, YJJ, and LYG performed the experiments. XYL, YL, KL, PPL, and XL assisted with the performance of experiments. XXZ, XSF and YJJ analyzed the data. XXZ and XW wrote the paper. All authors read and approved the final manuscript.
This study was approved by the Medical Ethical Committee of Shandong Provincial Hospital affiliated to Shandong University. All samples of patients were obtained with informed consent in accordance with the Declaration of Helsinki. All animal procedures described in this article have been approved by the Animal Care and Use Committee of Shandong Provincial Hospital affiliated to Shandong University.
Department of Hematology, Shandong Provincial Hospital affiliated to Shandong University, No.324, Jingwu Road, Jinan, Shandong, 250021, People's Republic of China
Xiangxiang Zhou, Xiaosheng Fang, Yujie Jiang, Lingyun Geng, Xinyu Li, Ying Li, Kang Lu, Peipei Li, Xiao Lv & Xin Wang
Shandong University School of Medicine, Jinan, Shandong, 250012, People's Republic of China
Xiangxiang Zhou
Xiaosheng Fang
Yujie Jiang
Lingyun Geng
Xinyu Li
Ying Li
Kang Lu
Peipei Li
Xiao Lv
Correspondence to Xin Wang.
Zhou, X., Fang, X., Jiang, Y. et al. Klotho, an anti-aging gene, acts as a tumor suppressor and inhibitor of IGF-1R signaling in diffuse large B cell lymphoma. J Hematol Oncol 10, 37 (2017). https://doi.org/10.1186/s13045-017-0391-5
Klotho
Insulin growth factor-1 receptor
Tumor suppressor


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Why is Youth Development a priority for Nestlé?
We believe that communities cannot thrive if they fail to offer a bright future for younger generations. As a major employer, Nestlé can make a positive difference. Our Nestlé needs YOUth initiative is helping young people around the world focusing on our core impact areas of employment and employability, agripreneurship and entrepreneurship. We offer training, mentorships and professional development opportunities and advance the skills they need to kickstart their careers. Building sustainable economies will depend on ensuring the next generation is able to access employment opportunities. Yet young people around the world are struggling to find such opportunities or to start their own business. This situation has only become more challenging as a result of the COVID-19 pandemic. In response, we scaled up the digitalization of our programs to ensure continued support for young people amid global restrictions. We are developing important new commitments to keep leveraging the momentum we have built to help young people reach their potential toward our 2030 commitment to help 10 million young people access economic opportunities.
Why do you believe that Youth Development is critical during this pandemic era?
According to the International Labour Organization, the impact of the pandemic on young people is deep and disproportionate, impacting their learning journeys and work, but also their mental health. Right now, millions of young people are under threat globally. For many, finishing their studies is out of the question; they will struggle to enter the labour market. Others have lost jobs and see no hope.
As we begin to recover from the pandemic, many businesses will lack the relevant skills needed to be successful in the future. That is why our contribution to the future generation is absolutely fundamental.
We cannot afford to lose a generation, at a time of technological changes with the sustainability imperative which is ours. We must skill, reskill and upskill our young people and we must do it quickly. It is the only way out of the crisis and the only path to sustainable growth.
What do you think differentiates our youth today from past generations?
Youth today have to be extremely agile, open minded and willing to challenge the status quo as a result of the rapid modernisation of our planet. In many cases traditional education, if available, no longer prepares young people for the current and future world of work. It is estimated that over 60% of the children entering primary school today will have jobs that do not exist yet. They need the right skills and experience to help them strive in a rapidly changing economic landscape. This is why since 2013 we have supported hundreds of thousands of young adults to prepare for the world of work by providing working opportunities, traineeships, internships and apprenticeships through our Nestlé needs YOUth initiative.
How can Caribbean youth be empowered to be leaders in their communities?
Do not be afraid. There are things you can do right now to prepare for the future — seek out every kind of experience, starting in your community: you will learn teamwork and leadership. Know that an exciting future lies ahead for you. There are big issues to solve. Some jobs will disappear, but new space will open up for more fulfilling work. Geography will not constrain your generation. People in previous generations had to be where the jobs were — thanks to the technology job opportunities can come to you wherever you are in the world. There is a lot to look forward to. Do not give up, be open, be daring and dream high.
Why should youth of the Caribbean attend the Nestlé Caribbean Youth Summit on Oct 5-6, 2021?
Under Nestlé's leadership, we have held Youth Summits all over the world in different regions, with thousands of young people attending and benefitting from this unique learning opportunity. This two-day virtual event in October will consist of motivational discussions, workshops, mentorship sessions and features from experts in various fields. Together with the Alliance for YOUth partnership, we will also make public our pledges to support youth development over the next three years. The Caribbean Youth Summit is meant to be fun and engaging, while thought provoking and challenging for all involved. More information about the Nestlé Caribbean Youth Summit can be found at Nestlé Caribbean Youth Summit | Nestlé (nestle-caribbean.com).
About Laurent Freixe
Laurent Freixe is currently Chief Executive Officer of Zone Americas, the largest region for Nestlé, accounting for 45% of its turnover. Born in Paris, France, Laurent Freixe joined Nestlé France in 1986. Laurent plays a leading role in promoting youth employment and employability across the company. Starting in Europe, Laurent launched the Nestlé Needs YOUth initiative in 2013 and has been successfully driving this global initiative that aims at providing a wide range of employment opportunities for young people under 30. This initiative purposes to strengthen their capabilities and professional skills, to raise their employability. In addition to his current responsibility, Laurent is International Youth Ambassador for the International Youth Organization for Ibero-America, Chairman of the Global Apprenticeship Network (GAN) and Chairman of the Global Alliance for Youth.