I had decided to continue looking at the Sacraments of Initiation on this blog. As readers know, I have been looking carefully at Baptism, the greatly misunderstood sacrament. Today, I am starting a mini-series on Confirmation, another misunderstood sacrament.
Part of the misunderstanding is simply the lack of good catechesis. In addition, many lay persons are stuck in magical thinking, about which I have written many times here. To receive a sacrament is not "to be saved", as in the Protestant idea of the fundamental option, either. See Veritatis Splendor for an update on that theological error. We have all the information we need online and in print to be educated Catholic adults.
Here follow a few points from the Church's teaching on Confirmation. And, I am reminded of St. Rita today, whose Secret in the 1962 Missal I have reads. "We pray thee, Lord, through the merits of Saint Rita, to pierce our hearts with the thorns of divine suffering; by the grace deliver us from all sin, and enable us to offer thee with pure hearts the sacrifice of praise through our Lord."
As in any of the sacraments, our hearts and minds are lifted up to God.
The CCC is a good place to start with some of the points relating to Confirmation. I cannot cover much today, but will write three or so posts this week. This is merely the first.
I am going to start a bit backwards, but as I was an RCIA coordinator and teacher in the past, this is where one starts. Not everyone can receive the sacrament of Confirmation. And, before all my Byzantine brothers and sisters chime in, let me say I agree one-hundred percent with the Sacraments of Initiation all being given together, in infancy, as done in the Eastern Rite. I wish we had not separated the Sacraments of Initiation.
Having said that and as this post in on the custom in the Latin Rite, let me continue by quoting the CCC.
So the CCC admits initiation in incomplete without Confirmation. One, however, must be baptized with a Trinitarian baptism and not one made "in the Name of Jesus", which is happening more and more in non-denominational Christian communities. One must check this out, as sometimes I have discovered in the past, that my candidate needs to back-track to baptismal preparation.
Very few Catholic parents realize that even before the traditional age of discretion, which is still seven in the Church, a child in the Latin Rite, under a threat of death, may be baptized.
This point must be emphasized, as the laity is confused here. Confirmation is not bar mitzvah or an adult appropriation of the Faith, as we must do that our entire lives. It is NOT a rite of passage. Confirmation is, again, a beginning sacrament for those who are being initiated into the Catholic Faith. We also underestimate the need for the virtuous life and the gifts of the Holy Spirit. In this time, children more than ever need sanctifying grace.
In diocese where I worked, Confirmation was at least a nine month, academic year process of teaching and initiation. Preparation cannot happen in a few weeks, as this is physically and mentally impossible, as well as spiritually dubious. Notice the last line, the community, that is the entire parish, but especially the family of the confirmand, takes responsibility for training. This cannot be left merely to the priest or religious education leader. The confirmand is part of a group which is practicing its Faith and is going out into that community to continue to feed that life of Faith in the community. I hear in England the phrase, "Oh, we lose the youth after Confirmation." If this is the case, either there was not enough preparation, or poor catechesis, or the youth were not in sanctifying grace to begin with and therefore, did not receive the graces of the sacrament. See below.
Perhaps this is my most important emphasis today. One must be in sanctifying grace to receive the sacrament. One must be a practicing Catholic. If one is not prepared to be a practicing Catholic and continue to be so after Confirmation, then the person is simply NOT ready to receive the sacrament. Again, there is no magic here. The sacraments are given without the framework of the life of the Church. One cannot receive grace in mortal sin and it is a mortal sin for a baptized Catholic not to be going to yearly Confession and weekly Mass on Sunday.
This point is simply ignored in many areas, and it is, frankly, an abuse of the sacramental preparation and an misunderstanding of the theology of the sacraments.
I know situations in parishes where a sponsor is literally grabbed by the priest or religious education coordinator before the Mass to stand as a sponsor. This is horrible. In some dioceses, there are sponsoring classes. and I when I was an RCIA coordinator, the sponsors came to the first few sessions with the candidates and came on one of the two retreats we had-one with those coming into the Church and the catechumens, and one which included the sponsors.
I understand the pressure from parents and in some parishes regarding Confirmation. But, I am reminded of Bonhoeffer on Cheap and Costly Grace. Either we do it right, or it is a travesty of our Faith. Confirmation is not a numbers game for a diocese to claim how many people have been confirmed in a year, nor is it a salve on the consciences of parents who just drop the kids off or sit in the car while classes are going on. I applaud the dioceses which have now included the parents in the Confirmation preparation. It is the only way, unless a young person, and I have known some as young as nine and eleven, come forward themselves for Confirmation and take the responsibility of living out the Catholic life after this sacrament.
and rituals at Sunday mass punctuate the transitions through the process.
This is the last day of my vacation in Kent. Out of the thirty-five days I have been here, the sun has shone five days, the fog has taken over the area about ten days, and the rain has pelted down thirty days or so. But, the worst natural phenomenon has been the wind. Gale-like winds are blowing even as I write this post. One cannot walk on the cliff paths in the wind. The wind howls off the sea and sweeps into the fields, pushing the seagulls into safe havens, as they move away from the shore. I must admit that wind bothers me as I grew up in a tornado alley. Wind is not friendly. It has a mind of its own. I feel like the seagulls, which have moved inland to get away from the blasts. One landed on the roof here and cried as if to say, I do not want to be here. I wonder what this large, handsome bird thought of the hail three days ago? Do birds think about the weather, or do these animals just react?
Now, I am the type of animal to think and to make the most out of any situation. And, I was here to visit with friends, old and new, which I did. I have had a great time.
However, as a walker, I have been disappointed. Last year when I was here in May, the sun shone almost every day and the weather was mild. Some of the locals called it "boiling", but boiling to me would be Iowa heat of around 95 hot degrees Fahrenheit in the shade, not a nice 75. It has been cold here this year, on the coast. I can see France at the edge of the drive, and it must have been cold there are well. In fact, as I write, it is 51 and it will get down to 45 tonight. But, the warm weather is on the way, so I am told.
Therefore, I did what I usually do when vacations meet bad weather. I read. As I am working my way through all the novels of Dickens, I managed one (out of the four since November 3rd). I am also re-reading E. M. Forster, some novels of which I taught years ago. I read two books on St. John of the Cross, and several others mentioned on this blog, and some not mentioned dealing with political theory, St. Benedict. the CCC, the Bible, of course, yet another short life of St. Therese, several encyclicals, and more.
I did not walk as much on the cliffs as I wanted to do, but perhaps, next year.


text:
Washington: 6:17 pm - January 26, 2023 - Jerusalem: 1:17 am - Shevat 5, 5783
Is any US, Israeli or world leader concerned about God's Biblical land plan?
By Bill Koenig
I wrote four letters of warning to President G.W. Bush and provided "Israel the Blessing or Curse" (in 2002) and "Eye to Eye" (in 2005). I sent a letter and "Eye to Eye" (in 2009) to the top Jewish members of Obama's administration — former Chief of Staff Rahm Emmanuel and Senior Advisor David Axelrod — and have written a letter and provided "Eye to Eye" (in 2017) to President Trump and his top advisors. The lobbying has been active behind the scenes.
Vice President Pence received copies of "Eye to Eye" from three different people. Secretary of State Mike Pompeo, Jarred Kushner and Jason Greenblatt received copies with personal explanation. They have all been informed and warned of the consequences of dividing God's covenant land. President Trump's administration has been more open to the warnings than any other administration, but they have decided to continue dividing the land and creating a Palestinian state.
Over 100 copies of "Eye to Eye" were delivered to U.S. Senators and U.S. House members.
In Israel, Benjamin Netanyahu received "Israel the Blessing or Curse" (in 2002) and "Eye to Eye" from me in February 2006 in Aventura, Florida, and the updated "Eye to Eye" in November 2018 from a very close friend of his in Jerusalem.
Ehud Barak and Ehud Olmert received copies from my assistant, Max, in Washington, D.C.
I personally gave copies of "Eye to Eye" to Ariel Sharon's son Gilad, Naftali Bennett, Speaker of the Knesset, Yuli Edelstein, and Moshe Ya'alon in Israel.
I gave "Eye to Eye" to top members of Israel's media: Caroline Glick, Barak Ravid, Udi Segal, David Horovitz, Steve Linde, Ariel Kahana, Danny Seaman, Jeremy Gimpel, Ari Abramowitz and others.
Has this effort made a difference? Netanyahu has said Israel is tired of managing the Palestinians. A few of my media friends in Israel said, "Bill, we hear you and understand what you are saying, but we don't want a million Palestinians descending on our streets."
Or they say, we don't want to have a Jewish minority in our own state. Some Israeli leaders believe the faulty demographics that say this will happen, when in reality the Jewish population is growing faster than the Palestinian population. Yoram Ettinger disproves the idea that Jews will become a minority in his report at https://theettingerreport.com/the-documented-jewish-arab-demographic-abo....
So, Israel and the region continue their move toward the day Jerusalem becomes a burdensome stone (Zechariah 12:3) and the final day conflicts led by Russia, Turkey, Iran and other Ezekiel 38-39 countries.
No American president has done more for Israel than President Trump; it isn't even close to previous presidents
I have mentioned this in many news reports;
U.S. Embassy move to Jerusalem;
Resolution backing Israel's claim to the Golan Heights;
Secretary of State Pompeo's stating Israeli settlements are not illegal, after a year of legal study;
National Security Advisor John Bolton warned the International Criminal Court against their isolation of Israel;
Ambassador Haley's chastised the U.N. Human Right Council hypocrisy and the U.N. Security Council Resolutions, that continually targeted Israel;
Stopped U.S. funding of the UNRWA refugee facilities and schools and their incitements against Israel;
Called for the Palestinians to stop funding terrorists who have killed Israelis;
Defense cooperation with Israel is at an all-time high;
On Iran:
Backed out of the Iran nuclear deal (JCPOA);
Approved the elimination of Iran's top military leader Soleimani, who was the architect of Iran's Middle East caliphate in Lebanon, Syria, Iraq and Yemen, who had planned a future major attack on Israel, and worked closely with Hezbollah;
Sanctioned Iran in order to stop their nuclear development program and missile development
Disruptive times when Israel's land is negotiated
There were major disruptive periods when President G.W. Bush was working on Israeli-Palestinian peace deals. I documented those in the first and second edition of "Eye to Eye – Facing the Consequences of Dividing Israel."
I provided specific examples of difficult personal, political and/or national matters for President G.W. Bush that occurred when he and his team were working on the two-state plan. Those periods would last anywhere from one week to six weeks.
President Trump has had one disruptive period after another. Most of those are not connected to Israel but some are; those disruptive periods were usually before and after the record hurricanes as previously noted.
No one has held the Palestinian leadership accountable for squandered billions of dollars:
Palestinians have taken in billions of dollars of aid from the U.S. and the EU. What do they have to show for it?
Gaza has turned into a full-fledged terror state.
At the Durban III event that I attended on the sidelines of the U.N. in September 2011, black scholar Dr. Shelby Steele, a senior fellow at Stanford University's Hoover Institution, compared the Palestinians to blacks who live by victimization and never take accountability for their failed community.
The failure to build vibrant Palestinian civil, economic, and cultural institutions has always been a significant obstacle to their dream of statehood — as surely as the Israeli people's success at doing so over the years preceding their declaration of statehood was a major asset.
Full text of Trump speech: This could be the Palestinians' last opportunity
https://www.timesofisrael.com/full-text-of-trump-speech-this-could-be-the-palestinians-last-opportunity/
Full text of Netanyahu's speech: Today recalls historic day of Israel's foundings
https://www.timesofisrael.com/full-text-of-netanyahus-speech-today-recalls-historic-day-of-israels-founding/
Trump – "Peace to Prosperity" Deal points
For Israel:
Israel to have full control of an undivided Jerusalem
The Palestinian capital of their state would be in East Jerusalem
Israel would have security from the Jordan River to the Mediterranean Sea
Israel allowed to annex 20 percent of the land in Judea and Samaria and swap some of its land in the Negev on the Gaza-Egyptian border
Existing Israeli settlements would be maintained; no Israeli removed from their homes; but would not be allowed to expand for four years
For the Palestinians:
A state of Palestine would be created that comprises up to 80% of the land of Judea and Samaria, and the new land in the Negev
The U.S. would open an embassy for Palestine and provide up to $50 billion in economic aid
The Palestinians would acknowledge Israel's right to exist, stop inciting terrorism, stop supporting families of terrorists, and disarm Hamas and Islamic Jihad
No Palestinian refugees would be resettled

redpajama_set_name: RedPajamaCommonCrawl


text:
Baharu railway station is a Kolkata Suburban Railway station on the main line. It is under the jurisdiction of the Sealdah railway division in the Eastern Railway zone of the Indian Railways. Baharu railway station is situated beside Baruipur-Kulpi Road, Uttarparanij, Baharu, South 24 Parganas district in the Indian state of West Bengal.

History
In 1882, the Eastern Bengal Railway constructed a -wide broad-gauge railway from  to  via Baharu.

Electrification
Electrification from  to  including Baharu was completed with 25 kV AC overhead system in 1965–66.

Station complex
The platform is well sheltered. The station possesses many facilities including water and sanitation. It is well connected to the SH-1. There is a proper approach road to this station.

References

Railway stations in South 24 Parganas district
Sealdah railway division
Kolkata Suburban Railway stations
Railway stations in India opened in 1882
1882 establishments in India
1882 establishments in the British Empire
Transport in Jaynagar Majilpur

redpajama_set_name: RedPajamaWikipedia


text:
Take Me Home Tonight è un film del 2011 diretto da Michael Dowse.

Trama
Siamo nel 1988. Matt Franklin è un giovane laureato al Massachusetts Institute of Technology che lotta per trovare la sua strada e lavora in un negozio al dettaglio specializzato nel videonoleggio.
Un giorno, Tori Frederking, la ragazza che amava al liceo, entra nel negozio per caso. Alla domanda su cosa ha fatto della sua vita, lui dice che è un banchiere della Goldman Sachs. Tori invita Matt a una festa organizzata in occasione del Labor Day da Kyle, il fidanzato di Wendy, la sorella gemella di Matt. Nel frattempo, Barry, venditore di auto e migliore amico di Matt, viene licenziato. Durante la notte, i tre protagonisti si troveranno ad affrontare i diversi aspetti del passaggio dall'adolescenza all'età adulta.

Produzione
Le riprese si sono svolte a Phoenix in Arizona nel 2007.

Titolo
Il titolo del film è ispirato alla canzone omonima interpretata da Eddie Money che non è, tuttavia, nella colonna sonora che comprende molte canzoni famose degli anni '80.

Il film è stato originariamente intitolato Young Americans e Kids in America in riferimento alle canzoni di David Bowie e Kim Wilde.

Collegamenti esterni

Film commedia
Film ambientati a Los Angeles
Film ambientati negli anni 1980


Happy May! Wheelhouse Detroit Bike Shop began operations on May 8, 2008. We are now entering our sixth year of business...in some ways, it seems like just yesterday that we opened our doors and, in others, decades ago! As that wise man Bob Dylan once sang, "I was so much older then, I'm younger than that now."
We'd like to sincerely thank you for being a part of the Wheelhouse family. We would not be here without you, and that's the plain truth.
We have been working hard to make this year our biggest and best ever and, therefore, this E-Blast might be our longest ever. Bear with us, we have some great news to share!
Sun - cruisers, comfort hybrids & adult trikes.
Origin 8 - Sun's urban offshoot; urban commuters, folding.
Brooklyn Cruiser - retro-styled cruisers.
Detroit Bikes - locally manufactured urban commuters (avail. in July).
Wheelhouse might be small in square footage, but we've become pretty adept at keeping a robust array of bike goodies on hand for all of your needs and wants. We stock a full range of top bike accessory brands such as Continental, Topeak, Planet Bike, Bell, Giro, Axiom, Saris, Kenda, Park Tool, Knog, Michelin, Timbuk2, Kryptonite, Master Lock, Pedros and Jandd. What do you need? A tube, tire, saddle, bag, basket, pump, mirror, helmet…We've gotcha covered! And if you're in the market for a sharp Wheelhouse t-shirt, you can grab one of those on your next visit, or snap one up online.
We recently installed a new POS system, which was an opportune time for us to review our service prices. Rather than merely increasing them, we've broken them down in a way that we believe to be transparent. There's no man behind the curtain here! Check out our full menu of service prices here. Please note that, in almost every case, service requires that you drop off your bike. We will do our best to let you know when it will be ready for pick-up. We all want to see you back riding as soon as possible, but to do things right can sometimes take some time.
We are excited to announce that we now offer Service Delivery! Yep, we can pick-up and/or drop-off your wounded cycle. For pricing, please visit our Service Page.
Our tour schedule for 2013 is finalized! A couple of notes worth mentioning: Our Underground Railroad Tour has added a stop at the Detroit Historical Museum to visit their recently renovated Doorway to Freedom exhibit; the Belle Isle tour has included a stop at the Acquarium to the roster; and we've developed a Music Heritage Tour that spans genres -- techno, rock, rap, soul, punk and Motown. You can scroll our Tours Page for details and to make reservations.
We also offer private tours, which are great as a corporate team-building exercise, non-profit fund-raiser, conference activity or pre-wedding excursion. A downloadable PDF with more info can be found here.
We are expanding our rental fleet, adding more cruisers, some comfort hybrids, an adult tricycle and an additional kids trailer. We will also soon have a couple of Autobikes! Online advance reservations available for a 10% discount.
GOT A GROUP? We offer discounts to groups of six to 20 with advance notice and a deposit. More info available at Wheelhouse Group Rentals and Schools & Scouts. We can also deliver bikes to your event; inquire at info@wheelhousedetroit.com.
We are also offering Cycle + Stretch with Midtown Yoga Shelter on May 19, register for that on our Tours Page.
A couple of annual rides that we are helping out with is the Ride of Silence on May 15 at 7 p.m. on Belle Isle and Cycle Into Spring on May 18 at Maheras Gentry Park.
Our Basic Maintenance Classes w/Fender Bender are back! This two-hour class will teach you some basic mechanics like changing a flat and go over some basic rules of the road and commuting tips. Classes cost just $5 and include a patch kit. Dates for the year: May 30 - Jun 13 - Jun 27 - Jul 11 - Jul 25 - Aug 8 - Aug 22 - Sep 12 - Sep 2.
As biking becomes more and more visible as a form of transportation, more and more events are adding bike parking as an amenity, and we are happy to help out! You'll find us at Detroit City FC home matches at Cass Tech; Detroit Tigers Bike to the Ballpark on May 22, and Riviere 28's Light up the Riverfront on June 6 and Soiree on the Greenway on July 18. More to come!
We're going to venture up to Port Austin for the weekend of June 7 for Detroit Up North - Take Two. More info about the 75- and 150-mile options, camping and more here!
Hope we see you soon -- and that we've given plenty of reasons for that to occur. Ride often and well.
The Week Ahead @ Wheelhouse!
Well, this week will sure keep us hopping over at the Wheelhouse. But that's cool, sleep is for the weak! Tuesday night at 6p.m., we'll have our Maintenance Class with Fender Bender! This 2-hour course is designed to get you comfortable making minor adjustments and repairs -- like fixing a flat -- and getting you on the road to regular commuting! It costs just $5 and you walk away with some knowledge and your own patch kit!
We are offering four tours this holiday weekend: Del Ray and Civil War History on Saturday; a long Techno route on Sunday; and a short Techno route on Monday. Last weekend, two out of three tours sold out, so make your reservation today! Find out more and sign-up at www.wheelhousedetroit.com/tours.
Monday, May 30, Memorial Day, we will be open holiday hours of Noon to 5 p.m. We will be closed on Tuesday, May 31 for employee observance of Memorial Day!
Read all about Bikes for Sale, our May tours and our upcoming Tuesday night maintenance classes in our May E-Blast. If you like what you read, head to www.wheelhousedetroit.com to sign up! Hello there, Wheelhouse friend!
Despite the glacial pace with which spring has approached Detroit, we are now fully ensconced in our seven-day-a-week summer schedule. We are open Monday through Saturday from 10 a.m. to 8 p.m. and on Sundays from 11 a.m. to 5 p.m. The only day we will be closed this month is May 31, when we'll take a belated Memorial Day vacay. Otherwise, come by during our posted hours and we promise we will be here!
Retail+Service You've got to swing by and check out the new bikes we have in stock! From Kona, check out the Dew, a light yet sturdy hybrid that retails for $429; the 2010 Smoke, a workhorse built for city streets, priced at just $360; the finely engineered Bike, which sells for $479; the 2010 Jake, a cyclocross monster priced at $764; and the utterly hip Paddy Wagon which, at $629 for the 2010 model, is ready to roll away. We also carry some classically styled cruisers from Sun Bicycles that start at $330 and streamlined urban steeds from Or igin-8 from $299. Also, we have one royal blue Sun Kruizer that we are willing to part with for $275.
For such a little shop, we've got a big shop personality -- we like to think! -- in terms of carrying a good variety of accessories that can make your bike prettier, tougher, more useful...whatever you dream it to be! We carry pumps, lights, racks, baskets, saddles, pedals, fenders, bells and helmets...What are you looking for? 'Cuz if it's not here, we can special order it with a quickness!
Tune-ups are back to regular price: 50 bucks. We pride ourselves on quality service and a quick turn-around, so drop your bike off and, when your pick it up, we guaran-sheed you will be satisfied. If not? Well, we will do our best to rectify that situation. That's how we roll.
Tour the city with Wheelhouse! We blast into our peak tour season with our May roster, which includes Hamtramck, Public Art, Architecture, Brewery History, Urban Agriculture, Eastern Market, Belle Isle, Automotive Heritage, Del Ray and Civil War History tours. We'll cap off the month with two different Techno in the 313 tours in partnership with the always-a-blast Movement Weekend! Be sure to visit wheelhousedetroit.com/tours to read up on our complete roster of offerings and to make a reservation.
We also offer private group tours, which are great as a corporate team-building exercise, non-profit fund-raiser, conference activity or pre-wedding excursion. Group rentals are also available at a discounted rate with an advance reservation. Give us a call at 313-656-2453 and we'll set something up!
New for 2011: Maintenance Classes Hot off the presses is our new collaboration with Fender Bender! We're teaming up to offer two-hour maintenance workshops on Tuesdays this summer where you can learn how to change a flat, patch a tube, adjust your seat and handlebars as well as some trouble-shooting tips, Rules of the Road and commuting guidelines. Classes cost just $5 and include a patch kit! Dates are May 10 and 24; June 7 and 21; July 5 and 19; and August 2, 16 and 30. They start at 6 p.m. sharp and are first-come, first-serve!
See you -- and the sunshine -- soon at the Wheelhouse. Right?


Just Wanderin': You Won't Believe Our Next Adventure !
You Won't Believe Our Next Adventure !
We've been in the Phoenix area for about a week now. When we first arrived the temperatures were running in the 90's but then it cooled right down into the 80's. Without humidity the weather has been perfect. However, sunshine wasn't the only weather we experienced. One day it rained hard and thunder and lightening cracked for hours. We think that's probably behind us now.
We've enjoyed connecting and re-connecting with people we haven't seen for a quite a while. Of course, that means we've had a lunch or dinner scheduled every day since we've been back. Our schedule has taken us around the Tempe, Mesa, Chandler, Apache Junction and even the Casa Grande area. We've also had a chance to go back to some of our favorite restaurants. We've shared lunch with the Dixons, the Hyders, the Rempes and the Kerseys. In addition to that we've also visited a few places with Terry's mom and sister.
We've also been busy transferring items from the Roadhouse to the RV.
After we were done, the little "Roadhouse" went to storage.
We've checked out quite a few RV parks in this area. The RV parks are definitely much lower than the $1000 a month we paid in Florida!! We found one in Mesa and in a few days we'll be settling in there for the next seven weeks.
We rode the kiddie train at the local park for something to do.
At first we thought our motorhome escaped any issues (other than dust) related to leaving it in storage under the hot Arizona. We discovered the batteries didn't fare so well and neither did the generator. The generator still won't start. So, we have an appointment to get it checked out. It's made us re-think again about leaving it all summer in the Arizona heat.
We've also visited Camping World a couple times. One was to make an appointment and another time was to sit in the waiting room while our trailer braking system was fixed. It seems it's been one issue after another.
We checked out the bargains at the Dillard's Clearance Center and found interesting shoes. None followed me out of the store though.
Interesting T-shirts were found at the Mesa Swap Meet.
Have you ever seen a Google Self-Driving car? Well, here's one for you. It did have a test driver with his hands on the wheel.
It seems we're going to become the caretakers of Buck, Justin's bulldog, for a few months. We discovered airlines won't fly Bulldogs due to their short noses. You might guess what the only option is of swapping Buck from Justin to us. It seems we have no choice but to drive.
We've been working on a plan and we think we've come up with one requiring the least amount of driving and the least expensive. When Justin gets his leave approved we'll drive our truck to El Paso and fly to Nashville or Knoxville. Drop off charges are quite a bit cheaper renting in Tennessee and dropping off in Texas. If we took it to Arizona, it would almost double.
After we rendezvous with Justin, the hard part begins and that's driving the rental car ALL THE WAY BACK TO EL PASO to pick up our vehicle before returning to Phoenix. That's the plan unless something changes. It's not a good plan but it's the best we can come up with for what we need to do. I hope Buck likes riding in vehicles because he's going to be doing a lot of it (and, so are we) !! I'm guessing in a few months we'll be doing this again.
What an adventure that is going to be ! Summing it up …. Justin is going to owe us BIG time for this!!! BIG TIME.
My goodness you never slow down, but as long as you are enjoying the life thats a good thing.
Have fun with Buck and good luck with your motorhome.
Since the shoes didn't follow you home, did one of those T-shirts?
WHAT? What the heck does a short nose have to do with flying? I don't get it. You sure are a couple of great grandparents taking care of Buck. I am not sure Paul and I would do the same.
We are looking for a place in Mesa/AJ. Where will you be staying. I know we need to start making reservations.
Holy cow, the fun never seems to stop. A dog for 3 months. Honestly you are wonderful grandparents. Buck doesn't shed much??
Hope if you venture back to Casa Grande you will let us know. Would love to see you!!!! Wow, you're really busy!! have one of those dog seat covers for the back seat if you would like it. We can't use it anymore.
Nice of you to "Buck Up" and take care of Justin's dog. I hope he appreciates all you're doing for him (Justin, not the dog).
I worry about our Journey now that it is going to sit in a storage lot for a while. I guess it would be smart to run it around every couple of weeks or so. It seems machinery likes to run and not sit.
Glad the temps have lowered for all your adventures in AZ. I always think those kiddie trains look like fun :-) The shoes, not so much. You never know what events will dictate plans, but Buck sounds like a good reason to take a little drive. How could you say no to that face???
Looks like a really nice rv resort to spend a few weeks!
Justin may be paying off that debt the rest of his life!


I have a confession. I have a serious love affair with sweet treats. Here's a quick and easy way to satisfy the craving for sweets without sacrificing your health to processed sugar and unwanted diary products. This creamy non-dairy ice cream will not disappoint you!
You will need: A food processor or a Vitamix ( a high powered blender works too)!
Add your coconut milk to your blender, then add your frozen bananas. Pulse for one minute then add your peanut butter and vanilla extract. Blend until smooth. Don't over-blend or the mixture will become too runny. You want a smooth creamy, ice cream texture.
Place your ice cream in a freezer safe dish and place in the freezer for at least 1 hour. I top my ice cream off with non-dairy dark chocolate shavings!


It may seem like common sense, but we really shouldn't run people over with our cars. Unfortunately there seems to be a lot of this going around lately and not just by Suge Knight. Using a car as a weapon is often a crime. This type of conduct would normally be charged as Assault With a Deadly Weapon under California's Penal Code Section 245. It could even be considered attempted murder or even worse if someone dies. There is only a legal defense if you are using reasonable and necessary force to defend yourself or another from an immediate threat.
Furthermore, if you hit someone and flee the scene afterward, you could be charged with felony hit and run causing injury – California Vehicle Code Section 20001. If there is serious injury, you could face years in prison. Also, fleeing the scene of an accident tends to make us look guilty, even if we are not. In the legal field it is what we call "consciousness of guilt". If the jury hears that you left the scene of a crime and you did this because you knew you were guilty they could actually hold this against you in trial. Remember, the law requires anyone involved in a traffic accident causing property damage or injury to stick around, identify ourselves and, if necessary, call for help.
In recent news, 25 year old Amir Bass allegedly hit his girlfriend's mother with his car, killing her, after an argument with her daughter. Sheriff's deputies arrested Bass yesterday on charges of ADW and Bass was said to have fled the scene after the incident which could give rise to additional charges. Deputies found Bass and his vehicle shortly after the incident and are investigating the tragic incident. Bass' girlfriend was treated and released at a local hospital. Bass was initially arrested on murder charges. Those charges were later downgraded to Assault With a Deadly Weapon.
Please remember that Amir Bass, Suge Knight and all others in this country are presumed innocent unless and until proven guilty beyond a reasonable doubt in court. Always exercise your constitutional right to remain silent when contacted by law enforcement. And finally, do not simply remain silent, but affirmatively assert your right to an attorney. Remember these words: "I want to talk to my lawyer". Because it is so important, I think I will say it again: "I want to talk to my lawyer". You can even get fancy and say: "I want to talk to my attorney". Whatever you say, just don't talk to the cops without your lawyer present.
If you are a suspect in a vehicular assault, ADW, murder or attempted murder or any other crime, call a veteran criminal defense lawyer who can help.


Three thousand years ago, in the territory now occupied by Juliaca and under Qaluyo culture influence, locals, farmers and guinea pig and llama breeders, built camellones or waruwarus to grow their food on the lake shores.
Other culture remains as Pukara, Tiahuanaco and Colla, which was defeated by the Inca Empire, during the Pachacutec and his son Mayta Capac reign, have been found in the area.
During colonial times, Juliaca was a distribution and in 1826, Bolívar named it as Villa de Comercio o de Tránsito (Trade or Transit Villa). Today it is a thriving city which industrial and commercial development, driven by informal gold mining, has become an attractive destination for workers from nearby areas.
Juliaca city looks, as its main architectural monument, the Santa Catalina colonial temple, built in the seventeenth century by the Jesuits.
Visitors can appreciate the Cristo Blanco, imposing statue made of reinforced concrete at the end of the last century, when they observe the city in a panoramic view from the Mirador de Waynarroque.
When you visit the Laguna de Chacas (Chacas Lagoon), located at 10 km in north Juliaca, you can enjoy a beautiful landscape and Iquinito hill, the highest on the Collao plateau with 4.565 meters above the sea level.
The Juliaca Carnival, organized in February, is one of the biggest tourist attractions in the city and the maximum expression of high Andean folklore. Streets, homes and businesses are decorated for receive the twenty-five thousand national and foreign musicians and dancers from the region, who parade long hours, up to 15 hours, the seven-day celebration.
Other tourist places: Catedral de La Ciudad de Puno, Balcón del Conde de Lemos, Museo Municipal Dreyer, Arco Deustua, Cerro Huajsapata, La Casa del Corregidor, Parque Mirador Puma Uta, Malecón Ecoturístico, Bahía de Los Incas, Reserva Nacional del Titicaca, Lago Titicaca, Islas Flotantes de Los Uros, Isla Amantaní, Isla Taquile, Sitio Arqueológico Inca Uyo, Complejo Arqueológico de Cutimbo, Complejo Arqueológico de Sillustani, Llachón, Buque Museo Yavarí.
Main hotels, as Suites y Hotel Don Carlos and Royal Inn have good international restaurants.
Another option is El Asador Point that opens from 6pm and is well known in the city for their grills, chicken, pastas and pizzas.
For lunch you can choose between the El Tablón poulterer"s restaurant and Restaurante Dominó, which offers regional and international cuisine.
Juliaca has shuttle service lines as Taxis Los Tigres (51) (051) 327272.
If you dare to try a different transport, the city is full of motorcycle taxis and tricycles, very popular to getting around.
Juliaca is known as the Windy City because winds are frequent and intense. The climate is generally dry and cold. The temperature ranges between 17 ° C and - 0.9 ° C. During the rainy season, between January and March, Juliaca presents hailstorms, snowfalls, thunders, and lightning, but rains are moderate.


When the thermostat's low by Emma Depanise and From outer freedom by Federico Federici, «Arkana», n.5, Cassie Hayes editor, 2018.
Emma DePanise has poems forthcoming or recently published in journals such as Superstition Review, Plume Poetry, Potomac Review, Nimrod International Journal, Little Patuxent Review and elsewhere. She is the winner of the 2018 Pablo Neruda Prize for Poetry. She currently studies creative writing at Salisbury University in Maryland.


The Nunchuk has similar motion-sensing technology to the Wii Remote, but also includes 2 buttons and a Control Stick. This allows you to move characters around while performing specific actions with the Wii Remote, like throwing a pass to your team.
Left handed? No problem. You can swap the Wii Remote and the Nunchuk from hand to hand for maximum control!


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Q: How to get limited number of fields from embedded document with mongodb-odm? I want to get the result of this mongodb command with Doctrine MongoDB ODM:
db.posts.find({_id: 3}, {comments: {$slice: [0, 5]}})

I was looking for it, but didn't find anything, neither in the documentation. If you have any idea, please help, thank you.
Here are my document classes:
/** @ODM\Document(db="dbname" collection="posts") */
class Posts{

    /** @ODM\Id */
    private $_id;

    /** @ODM\Field(type="string") */
    private $some_field;

    /** @ODM\EmbedMany(targetDocument="Comment") */
    private $comments = array();

    public function getSomeField() { return $this->some_field; }
    public function setSomeField($content) { $this->some_field = $content; }

    public function getComments() { return $this->comments; }
    public function addComment(Comment $comment) { $this->comments[] = $comment; }
}

/** @ODM\EmbeddedDocument */
class Comment {

    /** @ODM\Field(type="string") */
    private $timeStamp;

    /** @ODM\Field(type="string") */
    private $message;

    /** @ODM\ReferenceOne(targetDocument="Users", simple=true) */
    private $user_ref;

    public function getTimeStamp() { return $this->timeStamp; }
    public function setTimeStamp($str) { $this->timeStamp = $str;}

    public function getMessage() { return $this->message; }
    public function setMessage($str) { $this->message = $str; }

    public function getUser() { return $this->user_ref; }
    public function setUser(Users $user) { $this->user_ref = $user; }

}

A: While documentation on query builder's ->select() doesn't go much into details nor methods alike, following code will allow you to run your command:
$qb = $this->dm->createQueryBuilder(Posts::class)
        ->selectSlice('comments', 0, 5)
        ->field('_id')->equals(3);
$query = $qb->getQuery();
$results = $query->execute();

redpajama_set_name: RedPajamaStackExchange


The first phase of SAF Games qualifiers in athletics was held last week. It was an exciting two days of competition in which several national records were broken in the 400 Meters and High Jump. I really wish I had the chance to attend the event.
The Nepal Amateur Athletics Association would be wise start focusing on middle and long distance athletics events. Our geography and physique are perfectly suited to dominate those events. If Morocco, Kenya and Ethiopia can do it, why not us?
Our Karatekas are being forced to jump through hoops to attend the 9th AKF Championships in Guangzhou, China. The Nepal contingent are being asked by the Nepal Karatedo Federation to pay their own way once at the event. Also one of the Karatekas was denied a visa because she is Buddhist!


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Home Pakistan Sea World Show Is Coming to Pakistan: It's Bigger It's Better!
Sea World Show Is Coming to Pakistan: It's Bigger It's Better!
Maria Shafiq
The Sea World Show, with new mammals and exciting new tricks, is coming to Karachi soon, promising to be a treat! The immensely popular show will soon be hitting us with a bang, claiming to be much better and bigger.
There are not many family oriented entertainment outlets in the bustling city of Karachi -in fact, there is a dearth of safe and enjoyable places where families can break away from the usual routine of life and be able to sit back and relax.
Given this scarce number of options and a gaping void, you can now rejoice that the one-stop shop of entertainment, Sea World Show is paving its way towards Karachi! The Dolphin Show garnered popularity amongst teens, adults, and children last year and the Sea World Show is expected to receive an even warmer welcome.
[fb_embed_post href="https://www.facebook.com/SeaWorldShow/photos/a.1579509139009559.1073741828.1507296186230855/1580232885603851/?type=3&theater/" width="550″/]
The Sea World Show will feature new marine creatures such as the Black Dolphin, Beluga Fish and Sea Cats trained by none other than the famous Russian trainer, Vladimir.
The Sea World show is truly a unique mammal show, where humans get to interact with their favorite mammals. As of yet, there is no such arena in Pakistan where humans can actually intermingle with live animals and watch them perform startling tricks.
It opens a realm of enchantment and entertainment for the people of Karachi, to celebrate the thrill and adventure of live animal shows that are equally intriguing for children and adults, who just cannot seem to get enough of it.
The shows will be performed live at the Pakistan Maritime Museum. The timings, tickets and other details are to be announced very soon.
We'll just have to wait and see what new tricks have been learnt by these mammals, won't we?
Here's a clip of Vladimir, tamer of the sea creatures, wishing Pakistanis a Happy Independence day, that too in Urdu!
They are all set to take the excitement to a whole new level. We can't wait to watch these beautiful sea wonders!
Stay tuned to their Facebook page here for more announcements!


The 53rd Annual Faculty Research Lecture will be given by Professor Lise Getoor on Tuesday, February 26, 2019 at the Music Recital Hall in the Performing Arts Complex.
Data science is an emerging discipline that offers both promise and peril. Responsible data science refers to efforts that address both the technical and societal issues in emerging data-driven technologies. Prof. Getoor is a computer scientist who is well known for her theoretical work that integrates logic and probability to reason collectively and holistically about context in structured domains. In this lecture, she will describe some of the opportunities and challenges in developing the foundations for responsible data science. How can machine learning and AI systems reason effectively about complex dependencies and uncertainty? Furthermore, how do we understand the ethical and justice issues involved in data-driven decision-making? There is a pressing need to integrate algorithmic and statistical principles, social science theories, and basic humanist concepts so that we can think critically and constructively about the socio-technical systems we are building. In this talk, she will lay the groundwork for this important agenda.


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Home > Articles> Volume 1 >Whole-exome sequencing reveals genetic u…
19 Oct 2020 Volume 1 Views:2085
Shuhang Wang1*, Ning Jiang1*, Zicheng Yu2*, Yuan Fang1*, Shujun Xing1, Huiyao Huang1, Yue Yu1, Qi Fan1, Chao Zhang2, Zaixian Tai2, Jiucheng Liu2, Ruixian He3, Ning Li1
1Clinical Trial Center, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 100021, Beijing China.
2GenePlus-Shenzhen, 518000, Shenzhen China.
3Nursing Department, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 100021, Beijing China.
Correspondence: Ning Li (Clinical Trial Center, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 100021, Beijing China; E-mail: Lining@cicams.ac.cn). Ruixian He (Nursing Department, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 100021, Beijing China; E-mail: He87787591@126.com).
Asia-Pacific Journal of Oncology 2020, 1: 52-60. https://doi.org/10.32948/ajo.2020.10.15
Received: 17 Sep 2020 | Accepted: 10 Oct 2020 | Published online: 19 Oct 2020
Oral tongue squamous cell carcinoma (OTSCC) is a common malignancy, of which the incidence has increased in China in the last decade. Surprisingly, while multiple studies have revealed the mutational features of OTSCC in Western populations, limited data was shown in Asian patients. Herein, we utilized whole-exome sequencing to profile the genetic alterations in 13 Chinese OTSCC and compared them to those from 40 Western patients published in Cancer Discovery. In result, some key driver mutations were observed in both Chinese and Western cohorts, such as TP53 (Chinese 60.0% vs Western 60.0%), FAT1 (Chinese 7.7% vs Western 30.0%), CASP8 (Chinese 7.7% vs Western 10.0%) and NOTCH1 (Chinese 15.4% vs Western 10.0%), while mutations in CDKN2A (23.1%) and NTRK3 (23.1%) were only observed in Chinese patients, indicating these two novel mutations might play vital roles in OTSCC tumorigenesis specifically in Asian population. Mutational signatures depicted both common and distinct features across cohorts. In addition, significant copy number loss was found in 7q22.1, 9q13.1, and focal regions spanning CDKN2A and CDKN2B. FOXP1-TEX261 (2p13.3:3p13) fusion, reported in various cancer types, was firstly observed in OTSCC. Also, we identified numerous actionable mutations with FDA approved targeted. Taken together, our study revealed the mutational features of Chinese OTSCC patients, either similar or distinct to those of Caucasian patients. CDKN2A and NTRK3 were observed as two novel drivers that might play essential roles in tumorigenesis in Chinese patients, and were found as two potential therapeutic targets, rendering it promising to develop novel therapies.
Key words Actionable mutation, copy number alteration, oral tongue squamous cell carcinoma, whole-exome sequencing
Oral tongue squamous cell carcinoma (OTSCC) is a common malignancy arising in oral cavity [1], accounting for 354,864 new cancer cases and 177, 384 cancer deaths annually worldwide and is particularly prevalent in developing countries [2]. Given the different environment and living lifestyle across the world, different populations might have been exposed to different carcinogens, leading to distinct mutational backgrounds and molecular features underlying tumorigenesis [2]. To date, next-generation sequencing has unearthed key genetic alternations in Caucasian OTSCC population, including somatic mutations in canonical tumor suppressor genes and oncogenes like TP53 and NOTCH1 [3-5]. Nonetheless, limited reports have characterized the genetic underpinnings of OTSCC in Asian population. Herein by utilizing whole-exome sequencing, we scrutinized the mutational features of 13 OTSCC in China, shedding more light on the genetic mechanisms of OTSCC tumor progression in the Chinese population. We further integrated WES data of 40 Caucasian patients published on Cancer Discovery [5], exhibiting both common and unique genetic characteristics between Chinese and Western cohorts. Besides, novel actionable mutations were observed, rendering it promising to develop new targeted therapies for OTSCC patients.
Patients Enrollment, Sample Collection, Preparation and Ethics
13 patients diagnosed with OTSCC were enrolled from Cancer Hospital Chinese Academy of Medical Sciences from 2012 to 2019. Detailed clinicopathological characteristics of all patients were shown in Table S1. All patients underwent resection. Primary tumor tissues together with matched normal specimens were collected surgically and snap frozen in liquid nitrogen and stored at -80. DNA extraction was performed with TIANamp Genomic DNA Kits (Tiangen Biotech, Beijing, China) according to manufacturer's instructions. Written informed consent was collected in sample collection, gene sequencing and data publication. This study was approved by the Institutional Review Board (IRB) of Cancer Hospital Chinese Academy of Medical Sciences and conducted in accordance with the Declaration of Helsinki.
Library Construction and Whole-exome Sequencing
Library construction was performed using a custom 53M length capturing probe, made by Integrated DNA Technologies (IDT, IA, USA). Captured libraries were then pair-end sequenced in 100-bp lengths with Geneplus-2000 sequencing platform (Geneplus, Beijing, China) following the manufacturer's guidance. Reads were further mapped to the reference human genome (hg19) utilizing BWA aligner (version 0.7.10) for mutation calling after filtration.
Mutation Calling
Somatic single nucleotide variants (SNVs), small insertions and deletions (Indels) were detected using MuTect (version 1.1.4) packed in GATK (version 4.0). Variants with high frequencies among unrelated individuals across the world or specifically in east Asia were firstly filtered out as previously reported [6] and were listed in Table S2, then further filtered with the method demonstrated in a study [7] in order to screen out the key cancer drivers or cancer associated mutations. In brief, candidate alterations had to be mutations in known cancer associated genes collected in either Catalogue of Somatic Mutations in Cancer (COMSIC), OncoKB, CIvic (Clinical Interpretation of Variants in Cancer), CGI (Cancer Genome Interpreter) or IntOGEN (Integrative Onco-Genomics) database (all accessed in July, 2020). Mutations including nonsense, in-frame/frame-shift small insertions and deletions, and mutations in canonical splice-sites were kept. In regard of missense mutations, only those reported in COSMIC with a FATHMM-MKL (Functional Analysis through Hidden Markov Models) score of >0.5 (accessed in August 2020), or those identified in two or more of the following in silico functional analysis algorithms: predication score 0-0.05 in SIFT (Sorting Intolerant from Tolerant), "possibly damaging" or "probably damaging" in Polymorphism Phenotyping-2 (Polyphen2) or predication >0.5 in FATHMM-MKL were kept.
To study the significantly altered copy number of chromosomal segments, somatic copy number alterations (SCNA) was identified employing GATK (version 4.0) and GISTIC (version 2.0). An in-house algorithm was employed in detecting split reads and discordant read-pairs to identify chromosomal structural variation [6, 8]. Eventually, all candidate somatic variations were manually verified on the integrative genomics reviewer (IGV) browser to filter out the false positive.
To compare the mutational features between Chinese and Caucasian OTSCC populations, mutation data from WES of 40 Caucasian OTSCC patients, published in 2013 on Cancer Discovery [5] were downloaded from cBioPortal for Cancer Genomics database, with detailed clinicopathological information exhibited in Table S1. Mutations in genes collected in either COSMIC, OncoKB, CIvic, CGI or IntOGEN database were kept and subjected to following analyses. To find potential therapeutic targets, we mapped the unfiltered raw SNVs and Indels to OncoKB actionable mutation catalogue (accessed in July 2020), so as to avoid missing any potentially valuable candidates.
Analysis of Mutational Signature and Clonal Architecture
To find the possible mechanisms underlying OTSCC mutagenesis and reason the potential carcinogen-exposure or DNA damage repair deficiency of patients, the trinucleotide altering patterns of unfiltered somatic SNVs and fragment changing patterns of unfiltered Indels were matched to the known signatures described in COSMIC database using YAPSA (version 3.11). Contribution of each signature in samples was quantified and only signatures with contributions of more than 1% of all alterations were illustrated and considered in the subsequent analyses. Clonality of mutational events were inferred by looking at the variant allele frequency (VAF) ratio of mutations, calculated in the way as previously reported [9]. Specifically, VAF ratio was calculated by dividing the VAF of each mutation by the maximum VAF observed in the same sample. A higher VAF ratio suggested the respective event occurred at an earlier stage during tumor progression. Mutations with VAF ratios > 0.75 were determined as clonal mutations while the rest were considered as subclonal mutations.
Two-sided Mann-Whitney and Fisher's exact tests were performed on Graphpad Prism (version 7.01) or R (version 3.6.1) to generate the P value. All data processing, analyses and plotting were carried out utilizing R software (version 3.6.1) and/or GraphPad Prism software (version 8.0.2). Statistical significance was defined as P<0.05 for two-tailed student t-tests.
Mutation Landscape of Somatic Point Mutations
Whole-exome sequencing identified a total of 1,456 somatic SNVs and Indels in 13 Chinese OTSCC patients (CHN cohort) before filtration, while 3, 471 mutations in 40 Caucasian patients published in Cancer Discovery (CD cohort) were reported [5]. After filtration, 235 and 536 mutations in known cancer-associated genes were identified in CHN and CD cohorts respectively. In CHN cohort, genes with the highest mutation frequencies were TP53 (11/13, 84.6%), CDKN2A (3/13, 23.1%) and NTRK3 (3/13, 23.1%) (Figure 1A). Other recurrently mutated genes included ARHGAP5, CHD6, CNTNAP2, CSMD3, FAM47C, FAT4, GRIN2A, HCFC1, HERC2, KDM3B, KIAA1549, MUC4, NOTCH1, NOTCH2, ROBO4, SETBP1 and SETD1B, all mutated in 2/13 patients (15.4%) (Figure 1A). Mapping mutations into Reactome (accessed in September 2020) database suggested that mutations were enriched in NOTCH signaling pathway, RUNX3 signaling pathway, VENTX pathway and pathways involved in cellular senescence, which are closely related to cell cycle, apoptosis and stem cell phenotypes, and were shown to be associated with tumorigenesis in various cancer types [10-13]. In regard of CD cohort, recurrently mutated genes included TP53 (24/40, 60%), FAT1 (12/40, 30%), MUC4 (10/40, 25%), NEFH (7/40, 17.5%), PCLO (6/40, 15%), PIK3CA (6/40, 15%), HRAS (5/40, 12.5%), HLA-B (5/40, 12.5%), MLL2 (5/40, 12.5%), CASP8 (4/40, 10%), CHEK2 (4/40, 10%), HLA-A (4/40, 10%), MUC16 (4/40, 10%), NOTCH1 (4/40, 10%), SMAD4 (4/40, 10%) and ZNF521 (4/40, 10%) (Figure 1A).
We then compared the frequencies of mutations in known cancer-associated genes between CHN and CD cohorts (Figure 1B). In result, we observed similar mutational frequencies in some canonical cancer driver genes between two cohorts, such as TP53 (CD 60% vs. CHN 60%), FAT1 (CD 30% vs. CHN 7.7%), CASP8 (CD 10% vs. CHN 7.7%) and NOTCH1 (CD 10% vs. CHN 15.4%), indicating that they might play pivotal roles in OTSCC tumorigenesis. Notably, mutation frequencies of CDKN2A and NTRK3 in our cohort (3/13, 23.0% for both) were observed significantly higher than in CD cohort (0 for both; P = 0.012 for both; Figure 1B), suggesting CDKN2A and NTRK3 might be two novel drivers in OTSCC cancer progression among Chinese population.
To further justify the association of CDKN2A and NTRK3 with OTSCC tumorigenesis, we looked at the locations of their altered nucleotides on the protein-annotated chromosomes accordingly. As shown in (Figure 1C & Figure 1D), 2/3 missense mutations in NTRK3 located in the functional domains, namely immunoglobulin I-set domain and protein tyrosine kinase domain, whereas another mutation located in a non-functional region. As for CDKN2A mutations, 2/3 truncating nonsense mutations located in ankyrin repeats domain while another mutation located at a canonical splice site.
Mutational Signatures of Somatic SNVs and Indels in OTSCC
To gain an insight into OTSCC cancer etiology, we then matched the altering patterns of raw somatic SNVs and Indels to COSMIC single base signatures (SBS) and insertion and deletion signatures (ID) accordingly and quantified the contribution of each signature in both CHN cohort and CD cohort. In result, somatic SNVs were mainly attributed to three SBSs in COSMIC database, SBS39, SBS7b, SBS1 specifically, while Indels were attributed to ID1-14, ID16 and ID17 (Figure 2A).
SBS39, featured with predominant C > G alterations, was observed with drastically high distribution in head and squamous cell carcinoma and breast cancer and without known proposed etiology [14]. SBS7b, characterized by C > T mutations, was frequently found in skin cancer and was reported to be due to exposure to ultraviolet light [14]. SBS1, a clock-like signature with predominant C > T changes, is an endogenous mutational process initiated by spontaneous or enzymatic deamination of 5-methylcytosine to thymine which generates G: T mismatches in double-strand DNA, and was consistent with previous reports that SBS1 was one of the main signatures of OTSCC [15]. In our cohort, ID5 was observed particularly predominant, characterized by predominant single T base deletion, and was reported to be a clock-like signature associated with patients' ages [14].
We further quantified and compared the contribution of identified SBSs and IDs in CHN and CD cohorts to look at the difference in possible mutagenic exposure and DNA damage repair deficiency between Chinese and Caucasian populations. Interestingly, the exposing contribution of SBS1 was found significantly lower in our cohort compared to CD cohort (P < 0.001; Figure 2B), whereas SBS7b and ID5 were both observed with substantially increased contribution in our cohort (P < 0.001 for both) (Figure 2C & Figure 2D).
Clonal Architecture of Somatic Mutations in OTSCC
To look at the intra-tumor heterogeneity and infer the temporal order of the occurrence of somatic SNVs and Indels in Chinese OTSCC, we gauged the variant allele frequency (VAF) ratio of all alterations in our cohort with the method mentioned in previous literature [9]. Somatic mutations with higher VAF ratios were suggested to occur at earlier stages during tumor progression [9]. Mutations with VAF ratios greater than 0.5 were exhibited in (Figure 3), where black dots represent median VAF ratios when multiple mutations in the same genes were identified. Being consistent with the findings stated above, mutations in canonical cancer driver genes such as TP53 (0.78), CDKN2A (0.82) and CSMD3 (0.81) were determined as clonal mutations with VAF ratios greater than 0.75, indicating they might occur at an earlier stage compared to other subclonal mutations, and might play pivotal roles in OTSCC tumorigenesis. By contrast, some other canonical cancer drivers like KRAS, FAT1, NOTCH1 and APC were observed with relatively lower VAF ratios, suggesting they might occur at a later stage in tumor progression, exerting functions in tumor development and subclonal evolution. Subclonal mutations with various VAF ratios largely contributed to OTSCC intra-tumor heterogeneity and might be able to prime tumors towards drug resistant, disease relapse and metastasis.
Somatic Copy Number Alteration in OTSCC
To identify the recurrent focal somatic copy number alterations (SCNA), the GISTIC algorithm and the 2Mb windows setting were applied to our cohort. Among population, two significant regions of deletion, including 7q22.1 and 9q13.1, were detected (Figure 4A), while no significant amplified large segment was identified (Figure 4B).
We also manually checked the smaller focal regions of amplification and deletion of individual patients in order to characterize the driver genes significantly implicated in copy number changes. As a result, canonical oncogenes such as MYC, CCND1, FGFR1, ERBB2, PDGFRA, and KIT were gained or amplified whereas tumor suppressor genes such as CDKN2A, CDKN2B, and BRCA1 were lost or deleted (Table S3).
Chromosomal Variation in OTSCC
A total of 28 structural changes were identified in our cohort (10/13, 76.9%). Of particular interest, among 10 OTSCC patients harboring chromosomal changes, one case was paid special attention to and discussed in detail due to the detection of cancer associated FOXP1-TEX261 (2p13.3:3p13) fusion (Figure 5), which might exert potential functions in enticing tumorigenesis and were previously reported in other cancer types encompassing aldosterone-producing adenoma [16], prostate cancer acute lymphoplastic leukemia and etc [17-19].
Actionable Mutations of OTSCC in Chinese Population
In order to develop novel therapeutic interventions for OTSCC, we looked at the actionable mutations in our cohort by mapping unfiltered raw mutations into OncoKB actionable mutation catalogue (Figure 6), so that all potential candidate targets could be kept. Multiple actionable genes were observed with recurrent mutations, including CDKN2A, NTRK3, FGFR1, FGFR2, CCND1, BRCA1 and etc, functioning as potential candidates for developing novel targeted therapies. Of particular interest, CDKN2A and NTRK3 were observed mutated with the highest prevalence in Chinese cohort whereas no mutation in these genes was found in Caucasian cohort, suggesting they might play essential roles in OTSCC tumorigenesis and could be considered as two promising therapeutic targets, specifically in Asian population.
Figure 1. Somatic Single Nucleotide Variations and Small Insertions and Deletions Detected; A. Landscape of all mutations identified, panel at the top, mutation counts in each patient, right panel, number of patients harboring mutations in specific genes, panel at the bottom, source of patients, CHN- Chinese patients in our cohort (N = 13); CD-Patients published in Cancer Discovery (N = 40); B. Prevalence of Mutations in each cohort, * represents a P value < 0.05; C. Locations of sites of CDKN2A somatic SNVs/Indels in our cohort, Ank - ankyrin repeats domain; D. Locations of sites of NTRK3 somatic SNVs/Indels in our cohort, LRR8 - Leucine-rich repeat domain; I-set - Immunoglobulin I-set domain; Pkinase Tyr - Tyrosine protein kinases specific active-site signature domain.
Figure 2. Exposing Contribution of COSMIC signatures in 13 Chinese OTSCC patients; A. panel at the top, exposing contribution of SBS signatures; panel in the middle, source of patients, CHN- Chinese patients in our cohort (N = 13), CD-Patients published in Cancer Discovery (N = 40); panel at the bottom, exposing contribution of Indel signatures; B, C, D. exposing contribution of SBS1, SBS7b, ID5 in CHN vs. CD cohorts accordingly.
Figure 3. Mutations in genes with VAF ratios equals to or larger than 0.5 in 13 OTSCC patients.
Figure 4. Significant loss (A) or Gain (B) of copy number segments in 13 OTSCC patients. On each figure, the panel in the left represents the chromosome of each CNV segment located on; the numbers on the top represents the G-scores assigned by GISTIC for every cytoband plotted; the numbers on the bottom represents the -log10 transformed q values of each CNV.
Figure 5. TEX261-FOXP1 fusion identified in 13 OTSCC patients.
Figure 6. Mutations in Actionable Mutations by Searching OncoKB actionable genes in raw mutations of 13 Chinese OTSCC patients.
While incidence of head and neck cancer (HNC) has been gradually decreased in the last decades, several subtypes of HNC showed an increasing incidence, particularly for those occurred in oral cavity [20]. In this study, we illustrated the genomic landscape of OTSCC to dissect its genetic underpinnings. Albeit with a modest sample size, we integrated mutation data of 40 Caucasian OTSCC from a published dataset and validated the key findings of previous report as well as unearthed several novel drivers that might essentially contribute to OTSCC tumor progression. Lastly, we have shown that CDKN2A and NTRK3 together with some other actionable genes might be novel potential therapeutic targets, rendering it promising to develop new targeted therapies to prolong patients' survival.
Overall, compared to Western patients, the mutation frequencies of some known cancer associated genes in Chinese patients were quite close [5]. To this end, some key mutations in oncogenes and tumor suppressor genes such as TP53, PIK3CA, CASP8, NOTCH1, MUC4, FAT1 and etc were identified in both our cohort and CD cohort, proving their roles in OTSCC tumorigenesis as previously reported [5]. Also, our results suggested that the pathways these genes enriched in, NOTCH signaling pathway and CASP8 cellular necrosis pathway for instance, might be engaged in OTSCC cancer progression.
Notably, mutations in CDKN2A and NTRK3, both occurred in 23.1% patients in our cohort, were instead undetected in CD cohort, and both were categorized in OncoKB database as actionable genes, potentially offering novel targets for therapeutic development. Previously, CDKN2A and NTRK3 were reported to play key roles in various cancer types in the Chinese population including non-small cell lung cancer [21]. CDKN2A is a canonical tumor suppressor gene encoding p16INK4a protein, and was demonstrated to be a key driver triggering tumorigenesis in various cancer types [8, 22, 23]. The encoded p16INK4a protein exerts functions to orchestrate multiple signaling pathways in modulating cell cycle and senescence, through regulating the cyclin-dependent kinase (CDK) 4/6 and cyclin D complexes [8, 22, 23]. Moreover, genetic and epigenetic alterations in CDKN2A were exhibited to be indicative of disease relapse and dismal prognosis in cancer patients [8, 22, 23]. In our cohort, 2 truncating nonsense mutations and 1 mutation in canonical splice site were identified in 13 patients, and both 2 nonsense mutations located in the functional ankyrin repeats domain, which was shown to be involved in protein-protein interaction of p16INK4a to constrain cells from passing through cell cycle, and was suggested to play key roles in p16INK4a function [24, 25]. Our result indicated that mutations in CDKN2A might lead to the loss of p16INK4a regulatory capacity in modulating cell cycle arrest, and further result in OTSCC tumorigenesis. In addition, clonal CDKN2A mutation with high VAF ratio detected in our cohort suggested that CDKN2A might be altered at an early stage compared to other genes with subclonal mutations. Significant copy number loss of CDKN2A together with CDKN2B were also found recurrent in our cohort, which potentially resulted in a decreased expression level of p16 proteins, reducing its ability to maintain normal cell cycle arrest. Similarly, a recent study showed that CDKN2A copy number loss was observed highly prevalent (146/401, 36.4%) in HPV-negative head and neck squamous cell carcinoma, stringently associated with patients' dismal prognosis [26]. A germline polymorphism CDKN2B-AS1 (rs2151280 T>C) was also shown by another study to be associated with a higher risk of lung cancer [27]. Besides, differential expression in tumor tissues was found in various cancer types including hepatoblastoma, colorectal cancer and pituitary adenomas etc [28-31]. The down-regulation of CDKN2A and CDKN2B were found to associate with the recurrence of disease in oral cancer patients [32]. In 2003, Yakushiji et al showed that by comparing 25 pairs of primary oral squamous cell carcinoma (OSCC) and matched normal oral mucosa tissues, hypermethylation and down-regulation in both mRNA and protein expression levels were found in 48% samples, demonstrating a mechanism of CDKN2A undergoing hypermethylation, down-regulation to eventually trigger OSCC tumorigenesis [33]. In-vitro knock-in and knock-down assays of CDKN2A exhibited its basic functions in triggering G1 phase arrest during cell cycle in cancer cell-lines, where CDKN2A down-regulation was shown to be associated with enhanced cell proliferation [34, 35]. Besides, a CRISPR-Cas9 engineered CDKN2A knock-out model demonstrated depletion of CDKN2A facilitated melanocyte in-vitro motility and in-vivo invasiveness, indicating the function of CDKN2A to inhibit E2F1-mediated transcriptional activation of BRN2, which in turn to regulate down-stream signaling cascades to trigger melanoma metastasis [36]. Taken together, these results further solidified the conjecture that CDKN2A might be a novel cancer driver gene in OTSCC. The discrepancy of CDKN2A mutation in our cohort compared to CD cohort might be due to modest sample sizes of these two studies, while another possible explanation is somatic CDKN2A mutations might be specific to Asian OTSCC population whereas they occur less frequently in Caucasian population, awaiting more substantiation.
NTRK3 is another cancer suppressor gene regulating cell survival and apoptosis, encoding neurotrophin-3 receptor, which functions as a tyrosine kinase receptor to orchestrate multiple signal transduction cascades involved in cellular proliferation and was demonstrated to play important roles in various cancer types [37, 38]. Similarly, NTRK3 was found differentially expressed in various cancer types. Methylation and down-regulation were observed in tumors where reconstitution induced cell apoptosis, and a decreased expression of NTRK3 was reported to be associated with neoplastic transformation, cell migration and invasion in-vitro and in-vivo [39-42]. Aberrantly methylated NTRK3 was shown to be a biomarker of colorectal cancer [43]. Down-regulation of NTRK3 was suggested to increase cancer cell invasion, migration and proliferation [42, 44]. Moreover, ETV6-NTRK3 fusion was reported to be driver alteration triggering tumorigenesis of multiple cancer types, including secretory breast cancer, colorectal cancer and Spitz tumor [37, 45-48]. Recently, NTRK somatic mutations and ETV6-NTRK3 fusion were also reported in head and neck squamous cell cancers with a prevalence of 7.9% and 2.4% in Chinese patients (3/127; 10/127) respectively, and one of the ETV6-NTRK3 positive patient benefited from crizotinib treatment, offering an alternative treatment strategy [49]. Similar to CDKN2A, NTRK3 was observed recurrently mutated in our cohort, while no mutation was detected in CD cohort. 2 missense mutations of NTRK3 were found located in the immunoglobulin I-set domain and protein tyrosine kinase domain, leading to NTRK3 loss of function. Therefore, NTRK3 might also be another novel driver facilitating tumorigenesis and tumor development in OTSCC, and possibly specific to Asian population. However, ETV6-NTRK3 fusion was not detected in our cohort, possibly due to a limited sample size and/or a different sequencing strategy utilized in our study.
From our result, 76.9% OTSCC harbored mutations sensitive to off-the-shelf targeted therapeutic drugs, rendering targeted therapy a highly promising means of intervention to this malignancy. We manually checked the FDA approved medications in OncoKB database (Memorial Sloan Kettering Cancer Center) for these candidate genes. Among which, gefitinib, afatinib, osimertinib and dacomitinib were approved for EGFR+ non-small cell lung cancer, rucaparib was approved for BRCA1+ ovarian cancer, sunitinib, imatinib and regorafenib were approved for KIT+ gastrointestinal stromal tumor, and fulvestrant with alpelisib was approved for PIK3CA+ breast cancer etc. It's worth noting that even though these drugs were approved in numerous cancer types, limited numbers of drugs were developed or approved to be applied to OTSCC to date, therefore developing new therapeutic strategies for this cancer type might be of great significance to improve patients' treatment opportunities.
The mutational signatures of oral cavity cancer appear to vary depending on the exposure of environmental carcinogens [50], as habitual consumption of tobacco, alcohol, and betel quid (BQ) were recognized as the major etiologic factors for this type of malignancies. Previous literature has depicted the distinctive mutational features in tumors associated with cigarette smoking [51], alcohol drinking [52], and BQ chewing [53, 54]. In present study, we conducted a quantitative measurement of mutational patterns for our cohort and CD cohort. In result, though consistent signatures were identified between our cohort with previous report, significant difference was observed in contribution of mutational signatures between two cohorts. The discrepancy in signature distribution is considered reasonable given that the living environment and lifestyle largely vary across the populations.
Copy number alteration analysis revealed the deleterious somatic SCNA in hallmark genes of tongue cancer, such as amplification of MYC, CCND1 and FGFR1, and were also identified in previous reports [16, 55, 56]. In addition, high expression of ERBB2 was reported to exhibit important roles in the development of tongue cancer in patients with smoking history and/or alcohol consumption [57], and PDGFRA expression was significantly higher in oral cancer cohort with or without the establishment of tobacco risk factor [58]. Also as stated above, the down-regulation of CDKN2A and CDKN2B were found to associate with the recurrence of disease in oral cancer patients [32]. In conclusion, our SCNA findings were consistent with previous literature and have vital implications in clinically relevant applications.
Despite no overall structural variation pattern scrutinized, multiple chromosomal changes occurred in individuals might potentially contribute to cancer progression. In one patient, a FOXP1-TEX261 (2p13.3:3p13) fusion was observed (Figure 6). FOXP1 (Forkhead Box P1) encodes transcription factor Foxp1 expressed across the whole system, and was previously reported to exert functions in regulating immune response, cancer progression, organ development, and showed oncogene properties in acute lymphoblastic leukemia [19, 59-62]. As chromosomal changes involving FOXP1 was frequently found in various malignancies, FOXP1 served as a highly potential gene target for developing therapeutic interventions [62].
We reported the mutational analyses of 13 Chinese OTSCC patients. Apart from recently reported well-characterized alterations like TP53 mutation and CASP8 mutation in OTSCC, we also demonstrated several undescribed genetic changes such as CDKN2A, NTRK3 and FOXP1-TEX261 fusion. Also, several genes were identified as novel therapeutic targets, laying a solid foundation for developing new effective interventions.
The study was supported by the funder Chinese Academy of Medical Sciences(2019XK320068), and Chinese Academy of Medical Sciences(2020-I2M-2-007).
NL designed and supervised the study, RXH collect and preserve the patient samples. SHW, JN, ZY and YF analyzed the data and composed the article. SX collected clinical data and samples. YY, QF, CZ, ZXT, JL verified the data and methods. All the authors have read the article and were involved in the review and revision.
The authors declare that they have no conflicts of interest in this study.
The study was approved by the Ethic Committee of Cancer Hospital Chinese Academy of Medical Sciences (ID: NCC-002760).
Additional file 1: Table S1. Clinicopathological Characteristics of Patients Enrolled and in External Datasets.
Additional file 2: Table S2. Raw Mutations Identified from 13 Chinese OTSCC patients.
Additional file 3: Table S3. Significantly Implicated Driver Genes in Focal CNV Segments.
Prince S, Bailey B. Squamous carcinoma of the tongue. Br J Oral and Maxill Surg 1999; 37(3): 164-174.
Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018; 68(6): 394-424.
Vettore AL, Ramnarayanan K, Poore G, Lim K, Ong CK, Huang KK, et al. Mutational landscapes of tongue carcinoma reveal recurrent mutations in genes of therapeutic and prognostic relevance. Genome Med 2015; 7(1): 98.
Su S-C, Chang L-C, Lin C-W, Chen M-K, Yu C-P, Chung W-H, et al. Mutational signatures and mutagenic impacts associated with betel quid chewing in oral squamous cell carcinoma. Hum Genet 2019: 1-11.
Pickering CR, Zhang J, Yoo SY, Bengtsson L, Moorthy S, Neskey DM, et al. Integrative genomic characterization of oral squamous cell carcinoma identifies frequent somatic drivers. Cancer Discov 2013; 3(7): 770-781.
Li X, Huang H, Guan Y, Gong Y, He CY, Yi X, et al. Whole-exome sequencing predicted cancer epitope trees of 23 early cervical cancers in Chinese women. Cancer Med 2017; 6(1): 207-219.
Angus L, Smid M, Wilting SM, van Riet J, Van Hoeck A, Nguyen L, et al. The genomic landscape of metastatic breast cancer highlights changes in mutation and signature frequencies. Nat Genet 2019: 1-9.
Bartsch DK, Sina-Frey M, Lang S, Wild A, Gerdes B, Barth P, et al. CDKN2A germline mutations in familial pancreatic cancer. Ann Surg 2002; 236(6): 730.
Gerlinger M, Rowan AJ, Horswell S, Larkin J, Endesfelder D, Gronroos E, et al. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med 2012; 366: 883-892.
Allenspach EJ, Maillard I, Aster JC, Pear WSJCb, therapy. Notch signaling in cancer. Cancer Biol Ther 2002; 1(5): 466-476.
Ito K, Liu Q, Salto-Tellez M, Yano T, Tada K, Ida H, et al. RUNX3, a novel tumor suppressor, is frequently inactivated in gastric cancer by protein mislocalization. Cancer Res 2005; 65(17): 7743-7750.
Gao H, Le Y, Wu X, Silberstein LE, Giese RW, Zhu ZJ. Cr. VentX, a Novel Lymphoid-Enhancing Factor/T-Cell Factor–Associated Transcription Repressor, Is a Putative Tumor Suppressor. Cancer Res 2010; 70(1): 202-211.
Collado M, Blasco MA, Serrano MJC. Cellular senescence in cancer and aging. Cell 2007; 130(2): 223-233.
Alexandrov LB, Kim J, Haradhvala NJ, Huang MN, Ng AWT, Wu Y, et al. The repertoire of mutational signatures in human cancer 2020; 578(7793): 94-101.
Alexandrov LB, Kim J, Haradhvala NJ, Huang MN, Ng AW, Boot A, et al. The Repertoire of Mutational Signatures in Human Cancer. bioRxiv 2018: 322859.
Maitra A, Biswas NK, Amin K, Kowtal P, Kumar S, Das S, et al. Mutational landscape of gingivo-buccal oral squamous cell carcinoma reveals new recurrently-mutated genes and molecular subgroups. Nat Commun 2013; 4(1): 2873.
Takayama K-i, Suzuki T, Tsutsumi S, Fujimura T, Takahashi S, Homma Y, et al. Integrative analysis of FOXP1 function reveals a tumor-suppressive effect in prostate cancer. Mol Endocrinol 2014; 28(12): 2012-2024.
Ernst T, Score J, Deininger M, Hidalgo-Curtis C, Lackie P, Ershler WB, et al. Identification of FOXP1 and SNX2 as novel ABL1 fusion partners in acute lymphoblastic leukaemia. Br J Haematol 2011; 153(1): 43-46.
Sugimoto Y, Sada A, Shimokariya Y, Monma F, Ohishi K, Masuya M, et al. A novel FOXP1-PDGFRA fusion gene in myeloproliferative neoplasm with eosinophilia. Cancer genet 2015; 208(10): 508-512.
Chaturvedi AK, Engels EA, Anderson WF, Gillison ML. Incidence trends for human papillomavirus-related and -unrelated oral squamous cell carcinomas in the United States. J Clin Oncol 2008; 26(4): 612-619.
Wen S, Dai L, Wang L, Wang W, Wu D, Wang K, et al. Genomic Signature of Driver Genes Identified by Target Next-Generation Sequencing in Chinese Non-Small Cell Lung Cancer. Oncologist 2019; 24(11).
Foulkes WD, Flanders TY, Pollock PM, Hayward NKJMm. The CDKN2A (p16) gene and human cancer. Mol Med 1997; 3(1): 5-20.
Zhao R, Choi BY, Lee M-H, Bode AM, Dong ZJE. Implications of genetic and epigenetic alterations of CDKN2A (p16INK4a) in cancer. EBioMedicine 2016; 8: 30-39.
Borg Ak, Sandberg T, Nilsson K, Johannsson O, Klinker M, Måsbäck A, et al. High frequency of multiple melanomas and breast and pancreas carcinomas in CDKN2A mutation-positive melanoma families. J Natl Cancer Inst 2000; 92(15): 1260-1266.
Monzon J, Liu L, Brill H, Goldstein AM, Tucker MA, From L, et al. CDKN2A mutations in multiple primary melanomas. N Engl J Med 1998; 338(13): 879-887.
Chen WS, Bindra RS, Mo A, Hayman T, Husain Z, Contessa JN, et al. CDKN2A copy number loss is an independent prognostic factor in HPV-negative head and neck squamous cell carcinoma. Front Oncol 2018; 8: 95.
Lv X, Cui Z, Li H, Li J, Yang Z, Bi Y, et al. Association between polymorphism in CDKN2B-AS1 gene and its interaction with smoking on the risk of lung cancer in a Chinese population. Hum genomics 2019; 13(1): 58.
Al-Grawi EDC, Al-Awsi GRL. Expression of CDKN2A (P16/Ink4a) among colorectal cancer patients: A cohort study. J Pharma Sci and Res 2018; 10(5): 1145-1147.
Guled M, Lahti L, Lindholm PM, Salmenkivi K, Bagwan I, Nicholson AG, et al. CDKN2A, NF2, and JUN are dysregulated among other genes by miRNAs in malignant mesothelioma—a miRNA microarray analysis. Genes Chromosomes Cancer 2009; 48(7): 615-623.
Iolascon A, Giordani L, Moretti A, Basso G, Borriello A, Della Ragione F. Analysis of CDKN2A, CDKN2B, CDKN2C, and cyclin Ds gene status in hepatoblastoma. Hepatology 1998; 27(4): 989-995.
Simpson DJ, Bicknell JE, McNicol AM, Clayton RN, Farrell WE. Hypermethylation of the p16/CDKN2A/MTS1 gene and loss of protein expression is associated with nonfunctional pituitary adenomas but not somatotrophinomas. Genes Chromosomes and Cancer 1999; 24(4): 328-336.
Deepak Roshan VG, Sinto MS, Vargees BT, Kannan S. Loss of CDKN2A and CDKN2B expression is associated with disease recurrence in oral cancer. J Oral Maxillofac Pathol 2019; 23(1): 82-89.
Yakushiji T, Uzawa K, Shibahara T, Noma H, Tanzawa H. Over-expression of DNA methyltransferases and CDKN2A gene methylation status in squamous cell carcinoma of the oral cavity. Int J Oncol 2003; 22(6): 1201-1207.
Agarwal P, Sandey M, DeInnocentes P, Bird RC. Tumor suppressor gene p16/INK4A/CDKN2A-dependent regulation into and out of the cell cycle in a spontaneous canine model of breast cancer. J Cell biochem 2013; 114(6): 1355-1363.
Frost SJ, Simpson DJ, Clayton RN, Farrell WE. Transfection of an inducible p16/CDKN2A construct mediates reversible growth inhibition and G1 arrest in the AtT20 pituitary tumor cell line. Mol Endocrinol 1999; 13(11): 1801-1810.
Zeng H, Jorapur A, Shain AH, Lang UE, Torres R, Zhang Y, et al. Bi-allelic loss of CDKN2A initiates melanoma invasion via BRN2 activation. Cancer cell 2018; 34(1): 56-68. e9.
Wood LD, Calhoun ES, Silliman N, Ptak J, Szabo S, Powell SM, et al. Somatic mutations of GUCY2F, EPHA3, and NTRK3 in human cancers. Hum Mutat 2006; 27(10): 1060-1061.
Knezevich SR, McFadden DE, Tao W, Lim JF, Sorensen PHJNg. A novel ETV6-NTRK3 gene fusion in congenital fibrosarcoma. Nat Genet 1998; 18(2): 184-187.
Bu JY, LV WZ, Liao YF, Xiao XY, Lv BJ. Long non-coding RNA LINC00978 promotes cell proliferation and tumorigenesis via regulating microRNA-497/NTRK3 axis in gastric cancer. International journal of biological macromolecules. Int J Biol Macromol 2019; 123: 1106-1114.
Ge L, Li N, Liu M, Xu NZ, Wang MR, Wu LY. Copy number variations of neurotrophic tyrosine receptor kinase 3 (NTRK3) may predict prognosis of ovarian cancer. Medicine 2017; 96(30).
Luo Y, Kaz AM, Kanngurn S, Welsch P, Morris SM, Wang J, et al. NTRK3 is a potential tumor suppressor gene commonly inactivated by epigenetic mechanisms in colorectal cancer. PLoS Genet 2013; 9(7): e1003552.
Xiong D, Sheng Y, Ding S, Chen J, Tan X, Zeng T, et al. LINC00052 regulates the expression of NTRK3 by miR-128 and miR-485-3p to strengthen HCC cells invasion and migration. Oncotarget 2016; 7(30): 47593.
Tu J, Zhao Z, Xu M, Chen M, Weng Q, Ji J. LINC00460 promotes hepatocellular carcinoma development through sponging miR-485-5p to up-regulate PAK1. Biomed Pharmacother 2019; 118: 109213.
Li Z, Tognon CE, Godinho FJ, Yasaitis L, Hock H, Herschkowitz JI, et al. ETV6-NTRK3 fusion oncogene initiates breast cancer from committed mammary progenitors via activation of AP1 complex. Cancer Cell 2007; 12(6): 542-558.
Euhus DM, Timmons CF, Tomlinson GEJCc. ETV6-NTRK3—Trk-ing the primary event in human secretory breast cancer. Cancer Cell 2002; 2(5): 347-8.
Yeh I, Tee MK, Botton T, Shain AH, Sparatta AJ, Gagnon A, et al. NTRK3 kinase fusions in Spitz tumours. J Pathol 2016; 240(3): 282-290.
Lambros M, Tan D, Jones R, Vatcheva R, Savage K, Tamber N, et al. Genomic profile of a secretory breast cancer with an ETV6–NTRK3 duplication. J Clin Pathol 2009; 62(7): 604-612.
Xu J, Wang R, Wang T, Wang T, Shu Y, Gu D, et al. The Genomic Profiling in Chinese Head and Neck Cancer and Incidence of NTRK Fusion. J Clin Pathol 2020.
Beck TN, Golemis EA. Genomic insights into head and neck cancer. Cancers Head Neck 2016; 1(1): 1.
Alexandrov LB, Ju YS, Haase K, Van Loo P, Martincorena I, Nik-Zainal S, et al. Mutational signatures associated with tobacco smoking in human cancer. Science 2016; 354(6312): 618-622.
Chang J, Tan W, Ling Z, Xi R, Shao M, Chen M, et al. Genomic analysis of oesophageal squamous-cell carcinoma identifies alcohol drinking-related mutation signature and genomic alterations. Nat Commun 2017; 8: 15290.
Zhang W, Wang M, Wu Q, Zhu Q, Jiao Y, Zhu Y, et al. Mutational signatures and the genomic landscape of betel quid chewing-associated tongue carcinoma. Cancer Med 2019; 8(2): 701-711.
Su SC, Chang LC, Lin CW, Chen MK, Yu CP, Chung WH, et al. Mutational signatures and mutagenic impacts associated with betel quid chewing in oral squamous cell carcinoma. Hum Genet 2019; 138(11-12): 1379-1389.
Upadhyay P, Gardi N, Desai S, Chandrani P, Joshi A, Dharavath B, et al. Genomic characterization of tobacco/nut chewing HPV-negative early stage tongue tumors identify MMP10 asa candidate to predict metastases. Oral Oncol 2017; 73: 56-64.
Hammerman PS, Hayes DN, Grandis JR. Therapeutic insights from genomic studies of head and neck squamous cell carcinomas. Cancer Discov 2015; 5(3): 239-244.
Zhao Y, Fu D, Xu C, Yang J, Wang ZJOl. Identification of genes associated with tongue cancer in patients with a history of tobacco and/or alcohol use. Oncol Lett 2017; 13(2): 629-638.
Ong HS, Gokavarapu S, Tian Z, Li J, Xu Q, Cao W, et al. PDGFRA mRNA is overexpressed in oral cancer patients as compared to normal subjects with a significant trend of overexpression among tobacco users. J Oral Pathol Med 2017; 46(8): 591-597.
Barrans SL, Fenton JA, Banham A, Owen RG, Jack AS. Strong expression of FOXP1 identifies a distinct subset of diffuse large B-cell lymphoma (DLBCL) patients with poor outcome. Blood 2004; 104(9): 2933-2935.
Streubel B, Vinatzer U, Lamprecht A, Raderer M, Chott At. T (3; 14)(p14. 1; q32) involving IGH and FOXP1 is a novel recurrent chromosomal aberration in MALT lymphoma. Leukemia 2005; 19(4): 652.
Carlsson P, Mahlapuu M. Forkhead transcription factors: key players in development and metabolism. Dev Biol 2002; 250(1): 1-23.
Koon HB, Ippolito GC, Banham AH, Tucker PW. FOXP1: a potential therapeutic target in cancer. Expert Opin Ther Targets 2007; 11(7): 955-965.


text:
Public against alcopops election: poll
Tuesday, 11 August 2009 6:24PM
Distillers are stepping up their campaign against the alcopops tax hike, as the Rudd government prepares to have another attempt at pushing the measure through parliament next week.
The government plans to introduce legislation a second time for a permanent 70 per cent tax increase on ready-to-drink alcoholic beverages.
Legislation will also be introduced to validate the $365 million in revenue already raised by the measure.
The Distilled Spirits Industry Council of Australia (DSICA), which has waged an intensive campaign against the tax hike, released a survey on Wednesday conducted by Galaxy Research.
Almost two thirds of the more than 1,000 respondents believe a second rejection of the alcopops tax isn't a good reason to call a double-dissolution election.
"The Australian public does not think a second rejection of the RTD (ready-to-drink) tax bill is any justification for an early election," council spokesman Stephen Riden said.
Almost four in five respondents believe the tax increase should be abandoned in favour of a more comprehensive strategy to tackle binge drinking.
Federal Opposition Leader Malcolm Turnbull has argued Labor's move to resurrect its alcopops tax hike cannot trigger a double-dissolution election because the new bill won't be identical to the first.
The Senate rejected the government's initial bill validating the tax hike in March when Family First's Steve Fielding sided with the coalition and voted against the bill.
The federal opposition has indicated it will back laws validating collection of the alcopops tax to date, but it continues to oppose boosting the tax rate.
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Aerosols are tiny particles released into the atmosphere by human activities; they have proven negative effects on air quality—damaging human health and agricultural productivity. However, while greenhouse gases cause warming by trapping heat in the atmosphere, some aerosols can have a cooling effect on the climate because the aerosol particles cause more of the sun's light to be reflected away from the planet. Estimates indicate that aerosols have offset about a third of greenhouse gas-driven warming since the 1950s.
However, aerosols have a much shorter lifespan in the atmosphere than the gases responsible for global warming, resulting in an atmospheric distribution that varies by region, especially in comparison to carbon dioxide.
Now, a study by Geeta Persad and Ken Caldeira at the Carnegie Institution for Science finds that the impact these fine particles have on the climate varies greatly depending on where they were released. An open-access paper on their work is published in Nature Communications. . . .
This means that the degree to which aerosol particulates counteract the warming caused by greenhouse gases will likely decrease over time as new countries become major emitters.
The duo also found that there are significant regional differences when it comes to how strongly a country is affected by its own emissions.
Caldeira and Persad's work demonstrates that the climate effects of aerosol emissions from different countries are highly unequal, which they say means that policies must reflect this variation. . . .
I've been watching the BC-PNW fires (we're at Unhealthy for Sensitive Groups now, but it's been as bad as Very Unhealthy this past week with 1/4 mi visibility) and seeing the smoke waft over AB, SK and toward Greenland. This will likely mean increased black soot particulate falling on white snow/ice. Once the sun hits black particles on white snow, melting increases dramatically. I've personally observed this while hiking in the Cascades. Unfortunately, that was many decades ago when there was actually snow in the summer months. Now with all the fire danger and smoke, I'm not sure that I want to hike in those same regions anymore.


Back in November, I purchased the most beautiful skein of yarn with the intent of knitting it into a lovely, slouchy hat. But, feeling guilty about an unfinished knitting project (which also happened to be a birthday present for my sister), I set myself the goal of tackling it before even starting on my hat.
I FINALLY finished that project last week and set to work knitting my hat. It was so quick to knit up and I LOVED working with the yarn.
The pattern gave the option to knit 12 extra rows for extra "slouchiness" — I opted to leave those out, and I'm pleased with the results. If I make it again I might try adding the extra rows, just to see what it would look like.
To block the hat, I simply saturated it with cold water, rolled it in a towel to remove excess moisture, and pinned it in a circular shape on the floor. I dislike the ridge that so often accompanies beret hats, so I only let it block for a few minutes, then put it on my head to dry the rest of the way.
I only used about half of a skein of yarn, so I was able to get some fingerless mitts out of it as well. Maybe I'll show them to you someday.
Now I seem to have been bitten by the knitting bug. I can't wait to start on my next project!
What sorts of projects have you been working on lately?
I love the color you selected. It's gorgeous!
Very nice! The color is really pretty.
ADORABLE it looks soooooooooooooo cute!!!!!!!!!!!!!!!!
I absolutely love this hat! And the photos are fantastic. I have unfortunately seen it reposted on an instagram account (@_knitlife_) and they have not given you credit for the work nor the photo. They actually never give credit to the makers on any of their uploads, but yours is the only one I recognized. Just wanted to let you know! I hope they either take the photo down or give credit where it is due.


text:
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Игорь Вадимович Шипков (род. 21 марта 1985) — петербургский музыкант, певец, композитор, гармонист. Лауреат международного фестиваля «Балтика. Гармоника-98», фольклорного конкурса «Красная Горка», дипломант двух всероссийских фестивалей «Играй, гармонь!» (Иваново), гармонист золотой десятки Геннадия Заволокина (1999; дважды выступал в его передачах), лауреат фестиваля православной и патриотической песни им. Александра Невского (Санкт-Петербург, 2005), международного конкурса «Петропавловские ассамблеи», на котором занял 3-е место за исполнение музыки Баха и Моцарта на гармони.. Выпускник Санкт-Петербургской Академии культуры им. Крупской (2005). В Петербурге усилиями Игоря Шипкова был создан музыкальный центр «Играй, невская гармонь!», а на радиоканале «Петербург» он стал ведущим передачи «Завалинка»

Репертуар
В репертуаре более 60 произведений: Россия, Зимняя песня, Тихая моя родина, Мама, От печки, Я — деревня, Служить России, Русский парень, Галина, Никто тебя не любит так как я, Одинокая ветка сирени, Распустилась черемуха, Семеновна, 40 лет — бабий век, Играй гармонь, Крушина, Оренбургский платок, Гляжу в озера синие, Сельский парень, Заиграет гармошка, Любите гармонь, Коробейники, Эх деревня, Облепиха, Веревочка, Влюблённые трамваи (Белые-белые, красные, наши пути с тобой разные…) Поставьте памятник деревне, Шарманка и др. песни, а также наигрыши, участие в конкурсах частушечников, исполнение с залом попурри народных песен.

О нём
Отзывы из газет:
Виртуозный гармонист, певец, замечательный шоумен, — он с первых аккордов покорил публику…Его пальцы порхали по кнопочкам гармони, извлекая то задушевные, то искрометные плясовые мелодии, как русские народные, так и современные (Белгород).

… когда на сцену вышел высокий и статный, светлоликий Игорь Шипков, развернув меха своей гармони, и запел такую близкую сердцу песню Г.Заволокина «Я-деревне, я-село», а затем проникновенно исполнил песни на стихи Рубцова «В этой деревне» и «Тихая моя родина», лица зрителей просветлели. Гармонист вошёл в раж — приплясывал, стал петь частушки и припевки, и тут уже равнодушных в зале не осталось. Аплодисменты, крики «браво!». На ура восприняли его песню «Сельский парень». А в ответ на слова "выходи на пляску, кто в лаптях да валенках, у кого в душе радости огонь!"многие зрители вышли в проход и пустились в пляс. Когда же он исполнял попурри из народных и популярных песен подпевать начал весь зал. В едином потоке полилась русская песня (Великий Новгород).

Награды
 Член-корреспондент Академии русской словесности и изящных искусств им. Г. Державина.
 Золотая медаль «Г. Державин» за пропаганду в своём творчестве русского фольклора (от имени Академии русской словесности и изящных искусств им. Г. Державина).
 Медаль «60 лет полного снятия блокады Ленинграда» — «За пропаганду подвига жителей Ленинграда в годы Великой Отечественной войны».
 Лауреат международного фестиваля "Балтика. Гармоника-98″.
 Лауреат I степени международного молодёжного конкурса песни и танца народов мира и России (Санкт-Петербург, 2004).
 Лауреат международного конкурса «Петропавловские ассамблеи» (Санкт-Петербург, 2005).

Примечания

Ссылки
 Официальный сайт Игоря Шипкова
 Игорь Шипков. Профессионал. (интервью)
 Шипков Игорь Вадимович
 «Тверь влюбилась в гармониста» на портале eTver. 11 марта 2009 года.
 Татьяна Орехова. «Играй, невская гармонь!». 15 мая 2009 года.
 Константин Глушенков. Сыпь, тальянка, звонко…. Невское Время.


There's a whole generation out there that fondly remembers touring along with the Grateful Dead. Alas, with Jerry Garcia's death in 1995, the Dead, as they had been known and loved, were no more, but the spirit continues in the remaining members' projects and in the many tribute bands that crisscross Fairfield County.
This Friday, April 20, at 8 p.m. the freshly refurbished Wall Street Theater at 71 Wall Street in Norwalk, is hosting The Garcia Project. This band takes one specific show of the Jerry Garcia Band anywhere from 1976 to 1995 and recreates it, song by song.
If you can't make it to Norwalk, you can catch Legion of Jerry at BRYAC Restaurant & Raw Bar in Bridgeport. Located at 3074 Fairfield Avenue in Bridgeport, the show begins at 10 p.m. on April 20th.
And if it's more than music you're after, consider a trip to Greenwich's C. Parker Gallery which is presenting "From the Vault: The Art of Jerry Garcia and the Grateful Dead" from April 19 to April 29 at its 409 Greenwich Avenue location. This unique event includes original and hand-signed artwork from Jerry Garcia and band members Bill Kreutzmann and Mickey Hart, as well as personal items from Jerry Garcia on exhibit. Items will also be available for sale.
For information about the art exhibit and sale visit C. Parker Gallery . For tickets, to The Garcia Project's Norwalk show, which are $20 apiece, visit the Wall Street Theater . And visit BRYAC Restaurant for more info on its Dead-related event.