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Cusco To Machu Picchu 4 Days, Three Nights for only US $369 tour code MPT4/3
This is one of the most exciting adventure programs in the world, as you walk in the tracks of the original Inca Royal Trail, the same way and route the Incas did, ending up at the magnificent and legendary Inca citadel of Machu Picchu. This is a journey of foaming waterfalls, crisp, misty dawns and cloud-forests, bright afternoons and clear skies, a photographer's paradise.
Departures are daily except in the month of February. There is no minimum, but a maximum of 10 persons only is allowed. Passengers should plan to arrive in Cuzco at least one day in advance, in order to acclimatize themselves to the change in altitude.
Day 1 Cuzco/Chilca/Wallabamba Departure via overland vehicles from Cusco to our starting point where porters will be awaiting to start the trek along the turbulent Rio Vilcanota river, with stunning views of 19,000 ft. (6,300 m.) Nevada Veronica, to the ancient Inca city of LLactapata, with time to briefly explore the ruins. All the camp gear will be carried by porters, leaving you free to walk while carrying only a light pack with the essentials for the day's trek. Continue hiking along ancient Inca terraces up to the Cusichaca river to the village of Wayllabamba at 9,842 ft.(3,280m.) After a late afternoon arrival, make camp and overnight near the village.
Day 2 Wayllabamba/Pacaymayu- Begin trekking uphill slowly and steeply through an enchanted epiphyte laden misty-cloud forest to Warmihuanusca Pass, the highest pass at 13,750 ft. (4,580m.), but with magnificent views east to the Wayanay peaks and to the west, the rugged Vilcabamba range. Then we begin a downward descent to our camp for overnight.
Day 3 Pacaymaya / Winayhuayna- After breakfast, begin trekking a second pass at 13,100 ft.(4,350m.) and follow the ancient Inca highway (the original Inca Royal Trail) on the finest stretch of the trail. The Inca trail in this section is a white granite roadway, traversing a thickly forested mountainside. We camp here for overnight with magnificent views of Winayhuayna.
Day 4 Winayhuayna / Machu Picchu- Continue the trail along an ancient stone side-road, recently discovered but covered with orchids and hummingbirds, high above the Vilcanota River. Emerge at the "Gate of the Sun" for your first view of the extraordinary ruins of Machu Picchu from high above the city, and then begin the stroll down the "Royal Highway" into the lost city of the Incas. There will be time to thoroughly explore the ruins, and if so desired you can climb the nearby summit of Huayna Picchu for an incredible view from above, or relax at the ruins while contemplating the magnificent natural and manmade wonders. Return to Cuzco on the afternoon train, or spend an extra night (optional) here at Aguas Calientes village at the foot of Machu Picchu.
Regular Scheduled Departures
- every day, except February: US$ 369.00 per passenger -includes transfer
up to ruins, entrance fees to Machu Picchu and the Inka Trail fee.
IMPORTANT NOTICES AND CONDITIONS:The Program includes RT transfers in Cusco, all meals during the trek , two person tents plus a kitchen/dining room tent, all porters and cook, bilingual professional guide, emergency first aid kits, all entrance fees to Machu Picchu, and return train to Cuzco.
Not included: Air transportation Lima-Cuzco-Lima, airport taxes, nor any hotel while staying in Cusco. We suggest that all participants arrive at least one day in advance in order to acclimatize themselves to the altitude. We suggest you bring personal sleeping bag (we supply you with an air mattress), plastic rain poncho, sun screen or sun protector lotion, sun glasses, good trekking boots and thick socks, long pants and shorts, a light jacket, good sport socks, flashlight with extra batteries, any personal medications required, and plenty of film. Airport taxes are aproximately $9.00 per person and airfare Lima-Cuzco-Lima is around $170 is not included and are to be purchased seperately.
RATES AND CONDITIONS:PRICES:are in US$ per passenger. At the time of our confirmation of the rates and availability , we require your reconfirmation via email with your credit card information. There is no charge for cancellation up to 15 days before arrival. Cancellations up to 7 days before will be charged 50% of the tour price. Cancellations less than 7 days or "no shows" will be charged the full value of the tour. Rates are subject to change without notice however the rate we quote you by email IS GUARANTEED. Upon receipt of your credit card information your card will be charged the total price of the tour. You will receive a voucher via email for your tour. PLEASE, if you have to cancel advise us immediately to avoid uneccesary carges to your credit card. Do not hesitate to request additional information.
|We are proud of our association with American Express, Visa and Mastercard, companies known around the word. We are authorized to accept them for your reservation on our site.|
Reservations are handled by Viajes Pacificos and LatinTrip, Inc. fully licensed and accredited travel providers in Peru and in The United States, State of Florida, Seller of travel License #ST35323 located at 13876 SW 56 Street, #234, Miami, Florida 33175, toll free from the USA or Canada 1-800-811- 3077 from other countries call 1-786-272-9768 (in the USA), fax 1-786-312-1640, Email us at Reservations at Peru Hotels info@http://www.peru-hotels.com. The reservation center is open 9:00 AM to 6:00 PM East Coast USA Time, Mon to Fri, 9:00 AM to 2:30 PM Saturdays.


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Tax cuts are the government equivalent of "business expenses". People feel better in the short term but there is nothing to show for them in the long term. Tax cuts are like business expenses: money that is written off, to make the employees / voters feel better prior to an election.
No country, business, family or individual gets out of trouble by increasing their expenses. Spending for the sake of it, will not stop a recession or depression. The Americans got out of their last recession by investing in Public Works such as the Hoover Dam. Not by throwing money onto a burning building.
Tax cuts encourage people to waste money on cheap tat from China. The planet is in enough trouble without wasting more resources making rubbish we don't need.
The country needs investment, not more waste.
Public Works, and Public Education invest in the Nation's future. Better Public Transport, new roads, safe hospitals, better housing, but don't throw precious resources away on tax cuts.
Spend money, yes the country may soon need a boost. But please Gordon (NeoCon) Brown, don't run up more debt with nothing to show for it.
Investing in Public Works and Education and investing in this country's future YES!
Cutting taxes and other pea brained schemes spending money we don't have, on cake and circuses - NO!
Copyright (c) Dr. Liz Miller http://www.drlizmiller.co.uk

domain: finance


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In the first part of a three-part essay about design systems, I review two approaches for the creation of something equally as complex: a city.
Thoughts on Brasília, which I visited back in March.
Designed by Oscar Niemeyer, Brasília’s distinctive buildings feature a number of recuring design patterns and motifs.
For anyone coming to Brasília for its modernist architecture, no visit would be complete without a stay at this hotel. Designed by Oscar Niemeyer and opened in 1958, it hosted dignitaries such as Queen Elizabeth and Che Guevara before being devastated by fire in 1978. After facing decades of abandonment and neglect, it was modernised and reopened in 2006.

domain: artistic


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Pet/vet passport with all records
Fully vaccinated required kittens age
Information and care instructions
7 days a week support with all kittens needs
Delivery to any states available (not included to kitty price)
Check our feedbacks here:
A deposit of $500 is required to reserve a kitten. Deposit will place a hold on a kitten so it will not be available to anyone else. Deposit is non-refundable but may be transferred to a different kitten.
Final payment due the day your kitten arrives to his new home.
If you have any questions or concerns regarding payment please don’t hesitate to contact us.
We offer Word Cat Federation (WCF) pedigree.
Kitty can be spayed/ neutered on customer request
Kitty can be sold with breeding rights.
*Additional info pet request
Safe Buying Tips: We do not pre-screen advertisers. We do not offer any guarantees. Get the seller's phone number and call them. Use a payment method with buyer protection. Request recent vet exam documentation and call their vet. More tips

domain: pet


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But now, Australia’s very own Motown recording artist, Peter Cupples has embarked on such a challenge. Some people say, ‘if anyone can do it, it’s Cupples’, others shake their heads and call it ‘vocal suicide’. I guess we will have to let you be the judge of that.
Featuring some of the hottest musicians in Australia, this show celebrating the music genius of Stevie Wonder will absolutely delight Wonder fans and all those who love great music.
Stevie Wonder is one of the greatest artists of all time. Sometimes referred to as the eighth wonder of the world, Stevie is a true musical virtuoso and complete all-rounder. As a singer he possesses one of the most distinctive voices ever heard, his tone, control and range being second to none.
As a musician he could play piano, drums, harmonica and bass by the age of 10, and now he is proficient on so many instruments he’s regarded to be in a league of his own.
As a writer he’s penned and produced over 30 top ten singles, picked up 25 Grammies along the way and left us with some of the all-time classics: ‘You are the Sunshine of my Life’, ‘Superstition’, ‘For Once in my Life’, ‘Sir Duke’, ‘Lately’, ‘I Just Called to Say I Love You’, ‘All is Fair in Love’ just to name a few.
NOTE: MEMO Music Hall is compliant to all government COVID-19 regulations. The public health order now requires all patrons to be fully vaccinated and must show proof before entering our venue for this. Please have your certificate available for viewing prior to entry. We are unable to refund ticket holders who cannot show proof of full vaccination. Help us to keep music live.


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Former Maharashtra Ranji player Shekhar Gawli dies after falling into valley while trekking
Maharashtra Cricket Association also mourned his demise and offered condolences to his family.
Updated - Sep 2, 2020 5:39 pm
Former Maharashtra Ranji player Shekhar Gawli has been found dead after falling into a 250-feet deep gorge. Gawli’s body was recovered on Wednesday morning and the incident took place on the previous day when he had gone for a trek in Igatpuri, Nashik.
Gawli reportedly slipped during his trek and fell into the valley. Some reports also suggest that he was taking a selfie when he fell into the gorge.
Gawli was also the fitness coach of Maharashtra’s Under-23 cricket team. Gawli, a right-hand batsman and a leg-spinner, had also served as the assistant coach of the Maharashtra cricket team. The search for him began late in the night, with the local villagers also trying their best to find him. However, they were unable to do so and the search was halted at night. The body of Gawli was finally found on Wednesday morning.
Shekhar Gawli had represented Maharashtra in two Ranji matches
“His body was found at around 10 am on Wednesday. The body will be handed over to the family members after postmortem,” said an official as quoted by NDTV Sports.
Maharashtra Cricket Association also mourned his demise and offered condolences to his family. “In a tragic incident, our former player and current Trainer of Maharashtra team Shekhar Gawli passed away tragically. Our condolences to his family and friends. We are all with his family in this tragic situation. May his soul rest in peace,” Maharashtra Cricket Association said in a tweet.
In a tragic incident our former player and current Trainer of Maharashtra team Shekhar Gawli, passed away tragically .Our condolences to his family and friends .We are all with his family in this tragic situation.
May his soul rest in peace 🙏🏻
RIP Shekhar Gawli 💐#rip pic.twitter.com/q4QdSUIHJ6
— Maharashtra Cricket Association (@MaharashtraCric) September 2, 2020
Gawli, an all-rounder, had represented Maharashtra in two Ranji matches. He also had three wickets to his name.
BCCI, meanwhile, has announced November 19 as tentative date for the start of domestic season with Syed Mushtaq Ali T20 tournament. However, players contracted with IPL franchises won’t be able to play the initial phase of the tournament. The BCCI has also announced that there will be no Vijay Hazare Trophy, Duleep Trophy or Challenger Series this year and as of now, there is no provision for Irani Cup also.
Amid the COVID-19 pandemic, the domestic season will witness a day, meaning only Mushtaq Ali trophy and Ranji Trophy (December 13- March 10) comprising of 245 games will be played.

domain: sports


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There are various design options for your walk in shower. One of the most common variations comes in the form of a glass enclosure, with the glass helping create a bright and airy feeling. For an even lighter effect, consider adding a window to the shower, if it is possible. If not, try a skylight. If neither of these is possible, an all-glass enclosure would be better because it remains bright enough even with artificial lighting only.
You can also opt for walk in shower designs without doors. You could have the other walls solid, glass-only or a solid base with a glass top half. One thing to note about the doorless walk in shower is that it does not allow for much privacy, and this is generally true for all walk in showers. If privacy is not your priority (or is already provided by other means) then a walk in shower is perfect.
Each design comes with its own logistical and construction headaches on issues such as drainage. So make sure you consider these when choosing what style to go for.

domain: design


New Jersey News
Jewish businesses vandalized in Highland Park
HIGHLAND PARK, New Jersey (WABC) -- A man from New Jersey arrested for vandalizing several businesses owned by Jews could face hate crime charges.
The stores are all located in Highland Park.
Windows shattered at five businesses including a pizza joint and a gift shop.
The suspect allegedly threw cement from the sidewalk at the windows.
Police arrested 52-year-old Richard Green of New Brunswick.
Police have not revealed a motive.
Green is charged with criminal mischief but the charges may be upgraded to bias crime.
new jersey, vandalism, new jersey news
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- AccuWeather: Warmer days ahead
- Live: Eyewitness News on 7online streamed live!
- iWitness Photos and Videos
- Photos: Bus, taxi collide in Newark
- Target: 40 million accounts may be involved in breach 55 min ago
- US-India relations chilled over diplomat arrest in NYC
- Bronx cell phone store robber caught on camera 31 min ago
- Disturbing details emerge about suspected Etan Patz killer 17 min ago
- Rangel expected to announce run for 23rd term
- Jersey City boil water advisory in effect after main break
- Georgia winner of Mega Millions jackpot comes forward
- Most Fascinating People of 2013 revealed
- Photos: Most Fascinating People of 2013
- Target: 40 million accounts may be involved...
55 min ago


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Eva Mendes said that Cameron Diaz is very professional in her outlook and always gets the job done. Speaking at the Gala Premiere of The Other Guys, Eva said that Cameron Diaz is a good buddy to have as she never messes around with anyone"s life and always takes care of business. Eva also mentioned that she loves the way Cameron looks onscreen.
The actor also revealed that she has just finished directing her first short film which stars Christina Ricci and she would love to work in such similar projects in the future as well. Eva Mendes said that she never knew, she had a director hiding inside her and it is great to create a film from scratch. The actress is open to directing full length feature films in the near future. We hope this beautiful damsel"s dreams come true.

domain: celebrity


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Carbohydrates are nutritionally necessary nutrients that your physique makes use of to make power. And by having these available to burn, you will not then have to use proteins from your muscle groups for vitality. Nevertheless, it is also important to understand how the sugars in the carbohydrates are being saved in your body – are they being saved as glycogen or as fat?
o 2 pie crusts o 1 cup beer #1 Train Early ·Lucozade hydro – 10g Whole foods Whole grain brown rice, complete grain breads (natural or German type), sluggish cook plain oatmeal, Pink River Cereal. Leaving the skin on even in apple pies, gives a extra colourful consequence, and for the small ‘complainer’ who says one thing concerning the skin, simply inform them it’s healthier for them they usually can have a bigger piece of pie.
Recipe: What Else Preserves/Builds Muscle? 3) Seafood:
There are numerous illness you will get from foods that comprise certain levels of pathogens – ranges that wouldn’t usually hurt you. Nonetheless, when you’re pregnant you really cannot take any dangers with something that may or may not do you any harm. In spite of everything, you would by no means forgive your self if you picked up Toxoplasmosis or Listeriosis from meals that you had eaten and your child died or was born deformed as the results of this. However, this could happen and it is not a rare occurrence. The sorts of foods that could trigger harm are:
4) Beans and Legumes: Other examples of fruit and vegetables with these qualities underneath their pores and skin are: bell pepper, cucumbers, eggplant, grapes, green beans, nectarine, pear, peach, plum, yellow squash, zucchini. four – In giant bowl toss meat, apple, potato, cinnamon and sugar. 1 serving porridge – 14g
This would come with milk, cheese or yogurt.
Ultimately, getting a set of washboard abs is extra about removing fats from the stomach space to uncover the stomach muscle mass which are usually hidden underneath. You probably have already got strong abs, but nobody can see them, which is why nutrition is the important thing to learn how to great nice abs! All you want is someone to show you find out how to eat and exercise correctly.
2: 7-9 Servings of fruit and veggies Most cancers, along with coronary heart illness, diabetes, Alzheimer’s, and a growing list of different grievous ailments high the record of the inevitable outcomes of years and many years consuming mainly lifeless and worthless dietary substitutes for the real thing. o Surprisingly, but green-tipped bananas are higher for your well being than over-ripe bananas;
Begin with a great physical out of your midwife or MD. Ask your practitioner a few thyroid panel and Vitamin D level. Be sure your iron stores are good. If that weren’t unhealthy enough, MSG also damages the hypothalamus, the gland that controls appetite! Actually?

domain: food


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See why Philadelphia is the best city to learn sport, hospitality and tourism
STHM holds an intimate ceremony for the latest class of hospitality, tourism and sport leaders.
STHM professors and esport pioneers explore the $1.48 billion industry at the school’s first esport conference.
In just one week, Temple University raised more than $242,500 from 1,750+ donors.
Temple University’s School of Sport, Tourism and Hospitality Management professors and eSport pioneers explore the $1.48 billion industry at the school’s first eSport conference.

domain: news


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The Massachusetts Department of Transportation is moving towards all electronic toll collection on bridges, tunnels, and the Mass Pike. That means some drivers who got a free ride, won’t anymore.
The WBZ I-Team has learned a toll of at least 40 cents will soon be added to the ramps at the West Newton interchange of the Mass Pike. Currently, drivers could get on at exit 16, then get off at exit 17 in Newton for free.
This comes as MassDOT moves closer to all-electronic toll collection on the Tobin Bridge. The first toll booth has already been removed, and as soon as July 23, you’ll no longer be able to use cash on the bridge.
Vehicles without EZPass will have a photo of their license plate taken, and a bill will be mailed to the registered owner.
“It will make toll collection more efficient, more reliable. And in the end will increase the net revenue from the toll proceeds,” said MassDOT Commissioner Frank DePaola.
What do you think of the move to all-electronic tolling? Are you looking forward to the end of toll plaza backups, and will this encourage you to get an EZPass transponder? Share your comments below, and watch for them on WBZ News in the Morning from 4:30 to 7:00 a.m.


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Chilean cruise operators have sent a letter to the administrators of the port of San Antonio warning that if the intermittent service stoppages continue the companies they represent will have to take drastic measures. The current social turmoil and upheaval which has expanded to most of the country are distorting the normal functioning of the economy.
San Antonio is scheduled to receive some 25 cruise vessels this season, involving some 36.000 visitors and crew members and operators are increasingly concerned about the services provided by the San Antonio Puerto Central, PCE, and concessionaries.
The letter to the port authorities is signed by Victor Oelckers, General Manager of Inchcape Shipping Services, and addressed to Roberto Zilleruelo, PCE, CEO.
The letter is supported by the representatives in Chile of Holland America Line, Seabourn Cruises, Princess Cruises, Norwegian Cruises Line, Oceania Cruises, Regent Seven Seas, Aida Cruises, Cunard UK and Costa Crociere.
In the letter Oelckers points out to Zilleruelo the concern over the stoppage of port activities, which has had a very serious impact on the continuity of operations and the necessary safety for international cruise tourism.
Further it underlines that if new events of strikes take place in the future and they coincide with the dates of cruise calls, this could impede a normal and safe attention of passengers and crew members.
And if this scenario repeats itself, all signatories of the letter, will regrettably, have to consider and assess the suspension and elimination of San Antonio as a tourism destination, and hub port for the industry, with the negative and permanent consequences for the city and the country
Oelckers insists that the itinerary of the vessels was outlined two years ago and any modifications imply a very grave situation.
Any alteration or non compliance with the work programs already confirmed in your terminal, will mean, and without exaggeration, a total disaster, with permanent and catastrophic consequences, since they could mean the elimination of Chile from the cruise industry map, concludes the letter.
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Top rated Pizza Stones in 2019
I’m thinking about opening a small scale pizza parlor and I am now canvassing for the most affordable and high quality utensils and tools I need for the start-up business. I’m now looking into pizza stone products on the market and I realized I should check the internet first to see which brand is better. I found the best pizza stone reviews online and I have five of the best products to choose from. All of these have great quality and are from a trusted brand. I guess it would be the style that would matter the most.
If you think you can’t make a pizza at home that will taste like the ones from your favorite pizza house, think again. With the Old Stone oven pizza stone, you can definitely make the best pizza ever. It has the same material that kilns and furnaces have, doubling the effect of heating in old stone ovens, and producing a pizza crust that has the right amount of crisp and a very rich taste. Justifying the positive feedback from the best pizza stone reviews, this is indeed a great pizza stone.
Do you want to have a pizza stone that is lighter than most of the products on the market but has double the effects, producing delicious pizza crusts that has the perfect crisp and taste? The Emile Henry flame top pizza stone will surely help you make the best pizza ever. It features a Flame top technology which is intended for even heating and cooking of the pizza crust. The nonstick surface is perfect for easy cooking and cleaning. It is indeed the best pizza stone in 2019.
Unlike the normal circular pizza stone, the pizzacraft cordierite pizza stone has a square shape. It is made for heating and re-heating frozen pizza that is ready-made. Aside from making pizza, you can also use this baking stone for making bread and other pastries. You do not have to season or condition this stone. It works perfectly for a long time because it is made of high quality material. Because of that, it is considered as the best pizza stone in 2019.
If you are looking for a mix of great performance and an excellent stylish design, you need to check out the Good cook 4301 pizza stone. This is the perfect pizza stone for baking crispier and more delicious pizza crusts. It works well for heating and re-heating frozen or fresh pizzas. You can also use it for baking bread, biscuits, and other pastries. The stone is designed for even heating and there are absolutely no hotspots.
The GrillPro 98155 is a 13-inch pizza stone made of porcelain composite material. It is considered as a top rated pizza stone in 2019 because of its stylish structure and its high quality construction. The wire support is added for easy placing of the stone inside the furnace as well as for easy serving. It can resist cracking from temperature shock, making it a very durable pizza stone. You can use it for making pizza or breaking bread and pastries.


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Ahead of the 201st anniversary of Chile's independence and the Armed Forces Day, Mr. Joaquin Montes, Chile's Ambassador in Israel, held a reception at his residence on 18 September. Hundreds of businessmen, socialites, diplomats, military
attaches, friends of the embassy and Chilean expatriates attended the reception. The Ambassador noted the good relations between Chile and Israel in many aspects and wished the guests a happy new year. Minister Matan Vilnai delivered a speech on behalf of the Israeli government.
Photos by Diplomacy

domain: politics


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2023: Orji Kalu wins APC ticket for Abia north unopposed
Chief Whip of the 9th Senate, Senator Orji Uzor Kalu has won the ticket of the All Progressives Congress, APC for Abia north.
In the party’s primary election held on Saturday in Ohafia local government area of Abia State, Kalu who was also a former governor of the State ran opposed as two other aspirants to the office, Fabian Nwankwo and Ijeagu Emeaba voluntarily stepped down for him.
He however polled a total of 777 votes and was subsequently declared winner by Chairman APC primary election committee, Hon. Paul Uche.
The election which was conducted under the supervision of the Independent National Electoral (INEC), and monitored by the Nigerian Police Force (NPF), the Department of Security Service (DSS), and Nigerian Civil Defence Corps and other related security agencies commenced at about 11am with accreditation of delegates commencing at 11:58am just as voting began at 12:00 pm and was concluded by 1pm.
It will recalled that Kalu recently stepped down from the presidential contest which he had shown interest in.
With over 65 new community roads , water boreholes, construction and reconstruction of schools and hospitals, empowerments including 35 notable bills and motions to his credit, Kalu was expected to build on his achievements in his second term should he win in the 2023 general elections.


Republicans in the Arkansas House and Senate have just passed a bill that would ban physicians and other medical professionals from providing medically-necessary treatment and services to transgender minors. If Republican Governor Asa Hutchinson, who has already signed several anti-LGBTQ bills into law this month, signs HB 1570, experts warn transgender youths' lives will be at risk.
Calling it "one of the most extreme and harmful anti-trans bills in the country," the ACLU warned in a brief before the bill was passed that if signed, "it will be the most extreme piece of anti-trans legislation ever signed into law."
Lawmakers misnamed the legislation the "Save Adolescents From Experimentation (SAFE) Act." Monday's Senate vote was 28-7. The House bill was sponsored by Republican Rep. Robin Lundstrum, the Senate bill was sponsored by Republican Senator Alan Clark (photo).
"House Bill 1570 would prohibit healthcare professionals from providing or even referring transgender young people for medically necessary health care," the ACLU explains. "It would bar any state funds for gender-affirming health care for transgender people under 18, and it would allow private insurers to refuse to cover gender-affirming care for people of any age."
But that's not all.
"Doctors who provide medically necessary care to trans youth would risk losing their licenses and be subject to lawsuits by individuals and the state," the ACLU states.
ACLU attorney Chase Strangio:


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BRAF, NRAS and TERT mutations occur in more than 2/3 of melanomas. Its detection in patient’s blood, as circulating tumor DNA (ctDNA), represents a possibility for identification and monitoring of metastatic disease. We proposed to standardize a liquid biopsy platform to identify hotspot mutations in BRAF, NRAS and TERT in plasma samples from advanced melanoma patients and investigate whether it was associated to clinical outcome. Firstly, we performed digital polymerase chain reaction using tumor cell lines for validation and determination of limit of detection (LOD) of each assay and screened plasma samples from healthy individuals to determine the limit of blank (LOB). Then, we selected 19 stage III and IV patients and determined the somatic mutations status in tumor tissue and track them in patients’ plasma. We established a specific and sensitive methodology with a LOD ranging from 0.13 to 0.37%, and LOB ranging from of 0 to 5.201 copies/reaction. Somatic mutations occurred in 17/19 (89%) patients, of whom seven (41%) had ctDNA detectable their paired plasma. ctDNA detection was associated with shorter progression free survival (p = 0.01). In conclusion, our data support the use of ctDNA as prognosis biomarker, suggesting that patients with detectable levels have an unfavorable outcome.
Melanoma is the most aggressive type of skin cancer due to the high occurrence of metastases and resistance to conventional chemotherapy1. Moreover, melanoma rates have been rising for the last decades in predominantly fair-skinned populations and despite the favorable prognosis in early stages, local advanced or metastatic disease are still life-threatening conditions2. Currently, with the use of BRAF/MEK inhibitors and immunotherapy, the survival rates for metastatic melanoma have increased substantially. However, only a few patients produce durable responses, while the remainders develop resistance during treatment and experience disease progression at some point3,4,5,6.
The actual staging system from the American Joint Committee on Cancer (AJCC) includes prognostic and predictive biomarkers for melanoma7. However, AJCC only considers anatomical and histological features, which implicates that treatment decision and monitoring decisions are based solely on clinical and imaging findings, since there are no reliable blood-based biomarkers applicable for routine clinical use8,9. To rapidly identify metastatic progression and appropriately select patients for more aggressive therapy and avoid unnecessary treatments, there is a need for more tumor biomarkers.
In this context, a candidate biomarker that has been intensively evaluated is the circulating tumor DNA (ctDNA), which makes up part of cell-free DNA (cfDNA) from plasma10,11. The cfDNA comprises short fragments of DNA (160–180 base pairs), originated from cell death processes including necrosis and apoptosis and also cellular secretions12. The cfDNA concentration varies from 10 to 15 ng per milliliter of blood in healthy individuals, however it increases in certain conditions such as inflammation, exercise, tissue injury and in patients with cancer13. The ctDNA is the term for the fraction of cfDNA derived from tumor and therefore carries genetic alterations from the tumor. By the knowledge of tumor specific alterations, it is possible not only to identify molecular changes of the tumor but also to monitoring tumor burden in plasma. Recently, the detection of ctDNA alterations in melanoma patients has gain attention, due to its practical advantages: it is minimally invasive, allows serial collection, and is less prone to limitations such as quality of the material and tumor heterogeneity than traditional tissue biopsies14,15,16. Some relevant findings of this method include the association of ctDNA levels to tumor burden in metastatic melanoma patients and the ability of baseline values to predict outcomes11,17,18. The ctDNA variability during melanoma treatment is another possible tool for early assessment of therapy response or resistance18,19,20,21,22,23.
The most frequent and explored alterations are BRAF and NRAS mutations, present in 40–60% and 28% of patients, respectively24,25. In addition, a high frequency of mutations in the promoter region of TERT gene has been found in melanoma patients with a frequency ranging from 50 to 80% in several studies24,26,27, including Brazilian populations28. The most frequently described mutations are C228T and C250T, which are mutually exclusive24, but can co-occur with BRAF or NRAS mutations and are associated with lower survival28,29,30.
Therefore, we hypothesize that determining these driver mutations in tumor tissue and tracking them in plasma, it could contribute to identify melanoma patients at higher risk for progression. The aims of this study were to standardize a highly sensitive methodology of digital PCR (dPCR) to accurately identify hotspot mutations in BRAF, NRAS, and TERT in plasma and interrogate its feasibility as a liquid biopsy tool for melanoma patients.
Establishment of optimal conditions of digital PCR
For each BRAF, NRAS, TERT C250T and TERT C228T mutation detection assay, we performed temperature gradients to identify optimal annealing extension temperatures and times. Both BRAF and NRAS assays demonstrated better clustering between populations at 53 °C of annealing temperature, as shown in Fig. 1a–d. For BRAF mutation assay, the positive control (UACC62–BRAF V600E) showed two populations, the double negative droplets and the FAM positive (mutant) droplets, since the cell line harbor the V600E mutation in homozygosis (Fig. 1a). The BRAF negative control (SKMEL103 melanoma cell line), besides empty droplets, presented a population only on the y-axis, being correctly classified as wild type (Fig. 1b). For NRAS assay, the Q61R mutation positive control (SKMEL103) exhibited the mutation in heterozygosis, and therefore showed in the two dimensional plot representation of the four populations: the mutated homozygous droplets in blue, the droplets with the two alleles in orange, the wild type only droplets in green, and the double negative droplets in gray (Fig. 1c). As expected, the wild type control for NRAS (UACC62), presented double negative, and wild type droplets (Fig. 1d).
Due to the design characteristics of the oligonucleotides and amplification target region, both TERT assays required different cycling conditions. For TERT mutation the optimal conditions were under 54 cycles with annealing temperature of 59 °C (Fig. 1e). The positive control, heterozygous for TERT C250T mutation (SIHA cervical cell line), presented the four populations while negative control (HS587T breast cell line) presented double negative and the wild type droplets population (Fig. 1e, f, respectively). For the TERT C228T assay, despite extensive optimization attempts, the assay did not perform well in QX200 Droplet Digital PCR (Bio-Rad Laboratories, USA), presenting probe cross-reactivity. Instead, we benefit from testing TERT C228T in Quant Studio 3D (ThermoFisher Scientific, USA) equipment, using 54 cycles with annealing temperature of 55 °C (according to the manufacturer's recommendation). The positive control for TERT C228T (HS587T) had a similar distribution to the other heterozygous cell line previously demonstrated, with four populations (Fig. 1g). The wild type control (SIHA) presented some false-positive reactions (Fig. 1h).
Determination of limit of blank (LOB), limit of detection (LOD) and linearity evaluation
The limits of blank (LOBs) for each assay were calculated as described and results are summarized in Supplementary Table S1. For BRAF assay, since no mutant copies were observed in any control, the LOB was 0 copies/reaction. For NRAS assay, we found a LOB of 2.13 copies/reaction. Both TERT assays had a higher level of false-positive rates with LOBs of 3.995 and 5.201 copies/reaction for C228T and C250T, respectively (Supplementary Table S1).
The next step was to evaluate the linearity of assays and determine the analytical sensitivity of each assay. Using replicates of six dilutions of mutant in wild type DNA background, (Fig. 2). A high linearity between the expected and the measured fraction of mutant DNA was observed with a R2 = 0.9986 for BRAF (Fig. 2a), R2 = 0.9982 for NRAS (Fig. 2b), R2 = 0.996 for TERT C228T (Fig. 2c) and R2 = 0.9989 for TERT C250T (Fig. 2d).
The lowest concentration sample dilution was used to determine the LOD of assays, calculated as described by Armbruster and Pry31, varied from 3.71 to 9.76 copies/reaction with fractional abundance of 0.13% to 0.37% (Supplementary Table S1).
Patient demographics features and detection of baseline ctDNA
BRAF, NRAS, and TERT somatic mutations were evaluated in patient’s tumor tissue as part of clinical routine (Fig. 3). From 19 patients analyzed, 17 (89%) had at least one somatic mutation. Twelve patients had BRAF mutations (63.2%—10 patients: V600E, 2 patients—V600K), while two patients (10.5%) presented NRAS mutation (1 patient—Q61H; 1 patient—Q61R) and 14 patients had TERT alterations (74%) (7 patients—C228T; 7 patients—C250T). Most patients (n = 11) presented more than one gene altered, and as expected BRAF and NRAS were mutually exclusive. Two patients were triple negative for selected genes (IDs 4 and 14) (Fig. 4).
Clinical information of the nineteen patients are summarized in Fig. 4. Overall, 12 (63.2%) patients were female and 7 (36.8%) were male, the ages ranged from 34 to 79 years, with a mean of 61 years, 15 (79%) had distant metastases (stage IV) and 4 (21%) presented only lymph node involvement (stage III). The most frequent histological subtypes were acral and nodular melanoma (5 patients each). Two patients (IDs 4 and 9) had only brain metastasis with unknown primary site and 3 patients had no information on the primary lesion, being classified as an unknown histological subtype. Regarding the Breslow thickness of the primary lesion, both the mean and the median thickness of the group were around 7.5 mm, so that the patients were grouped according to this value, above and below (Fig. 4). Finally, 5 patients died and 14 are still being followed up.
ctDNA could be detected in seven (41%) of the 17 patients that had at least one mutation identified in the tumor tissue. Quantification of mutant targets (copies/reaction) for each assay is described in Supplementary Table 1 (Table S2). Analyzing by gene, BRAF mutations were detected in plasma from in 5 of 12 cases (41.6%), in 1 of 2 cases of NRAS positive (50%), and in 5 of 14 TERT positive mutations (35.7%). From 12 patients that carried two genes mutated on tumor tissue, both alterations were detected in plasma in 4 cases (33.3%), in 7 patients, neither mutation was detected in ctDNA, and 1 patient had only one of the mutations observed (8.3%). Patients without mutation in the FFPE tissue did not exhibited mutation in the plasma (Fig. 4).
Association of ctDNA and patients’s prognostic factors
We grouped the 17 patients according to ctDNA detection in plasma: detectable (n = 7) and undetectable (n = 10). The association between the clinical features of the 17 patients and ctDNA detection is shown in Table 1. Of the patients with detectable ctDNA, 5 (71%) were younger patients (< 61 years). It was observed a tendency between Breslow thicknesses of primary lesions greater than 7.5 mm with detectable ctDNA, yet it was not significant. Also, a higher frequency of detectable ctDNA was observed in patients with more than one metastasis site. Individuals with isolated central nervous system (CNS) metastases had no detectable ctDNA (IDs 2 and 9).
Association of ctDNA and patient outcome
The median follow-up of patients was 130 days, ranging from 7 to 253. Patients who had detectable levels of ctDNA had a shorter progression-free survival when compared to patients with undetectable mutation (p = 0.01) (Fig. 5). The median time in days for progression in patients with circulating tumor DNA was 50 days, and in patients without circulating tumor DNA it was 146 days. Patients with detectable ctDNA had significantly increased risk of progression compared with those with undetectable ctDNA (HR: 6.38; 95% CI 1.26–32.1; p = 0.025).
The emergence of target therapy and immunotherapy has contributed significantly to the survival of patients with metastatic melanoma4. However, melanoma lacks prognostic and predictive biomarkers, which implicates on the challenges in select patients with a poorer prognosis that will benefit from a more aggressive therapy and for early identification of disease progression.
Here, we validated a dPCR approach that detects the most frequent melanoma alterations in cfDNA in a real-world context. We demonstrated that these specific mutations are present in tumor tissue of 89% of our patients, and taking together, mutations could be detected in plasma from seven of seventeen patients (41%). Briefly, ctDNA was detected in 5 of 12 cases BRAF positive (41.6%), in 1 of 2 cases NRAS positive (50%), and in 5 of 14 TERT positive mutations (35.7%).
Liquid biopsy appears as a minimally invasive model, low-cost, sensitive, reproducible and with a wide variety of applications in personalized medicine for melanoma through analysis of circulating tumor DNA11,25,32. In attempt to implement liquid biopsy in a clinical setting, our group sought to standardize a platform to evaluate hotspot mutations in melanoma patients. In addition to BRAF and NRAS alterations, which are the most explored genes in most studies33, we included the TERT gene, since mutations in its promoter region have been described by its high frequency in melanoma patients27,28,29. The dPCR methodology was chosen due to its high sensitivity and accuracy, especially in the detection of rare alleles at low frequencies, that best fits in this context34,35. The major disadvantage of dPCR is that, generally, only one target per test can be evaluated in dPCR, which implicate higher cost and time consuming, when analyzing a large number of genes. Even that multiplex assays have been developed recently, there is a limitation in number of targets to be included, due to the necessity that PCR conditions must be the same for all targets. Besides that, the majority of dPCR systems have only two channels of fluorescence detection (FAM and VIC/HEX), which can limit the design of assays with many targets.
Other methods of ctDNA detection can be used, as Next-generation sequencing (NGS). NGS can provide not only the ability to detect genome-wide DNA variation but also the use of multiple targeted panels36. The use of these panels has lower cost compared to whole-genome-sequencing (WGS) and whole-exome sequencing (WES) and also offers the advantage of reducing the DNA input and still achieving a high sensitivity. However, the detection of rare alleles at a low frequency requires a very high coverage (> 10.000×), which increases sequencing cost37. In addition, there is an important limitation in the analysis of TERT promoter mutations in NGS panels. Despite validated for FFPE NGS panels38, TERT promoter mutations have not performed well for cfDNA in NGS custom panels39,40 due to its high GC content, and so, it is mostly described in droplet digital PCR approaches41,42. Furthermore, BRAF and NRAS mutations are mutually exclusive, as well as TERT C250T and C228T, and then, a maximum of two mutations will be tested in plasma for each melanoma patients, making feasible the dPCR approach proposed in this study.
We detected plasma ctDNA in seven of seventeen patients who had at least one somatic mutation in tumor tissue. This is in agreement with three other similar studies using ddPCR, which indicated that ctDNA detection rates are associated with tumor burden18,23,43. McEvoy et al18 demonstrated a detection rate of 71.8% in a cohort of 32 stage IV patients. In keeping with the relationship to burden, detection rates fell to 12% in lower stage patients (EC II and III)23. The same group showed a detection rate of 53% at baseline in stage IV patients that undergone treatment with PD1 inhibitors22. Data from Braune and colleagues, showed an even higher detection rate of 87.1% in a group of 62 patients that comprised 50 stage IV and 12 stage III melanoma patients43. The fact that our group comprised both stage III and IV indicates that our detection rate was similar to other studies examining these stages. However, the study from McEvoy, demonstrated that ctDNA detection not only depends on the clinical stage, but also on the metabolic tumor burden (MTB) and metabolic tumor volume (MTV) accessed by 18F-labeled fluorodeoxyglucose positron emission tomography/computed tomography, as ctDNA was undetectable in all patients with an MTB value of ≤ 1018.
Another important aspect associated to ctDNA detection is the volume of plasma processed and the input of DNA in dPCR reaction, as the screening of small amounts of sample may not always represent the true mutant content, mainly when they are present in a few copies in our sample. The high detection rate from Braune et al. might be associated to the higher input screened. Despite using 2 ml of plasma they tested samples in 4 replicates43. Corless and colleagues screened 12 replicates from plasma samples for TERT evaluation in ddPCR42. Some other studies isolated DNA from a larger volume of plasma, up to 5mL41,44. We presume the most important limitation of our assay was the fact that it was possible to used only 2 mL of plasma and 10 ng per reaction, which generated a higher LOB and, consequently LOD. Higher cut-offs implicates in lower sensitivity. It can have affected the accuracy of our experiment in diagnostic sensitivity, but it demonstrated to be important for determining patient prognosis.
TERT mutation occurs in up to 80% of cutaneous melanomas, which makes this gene extremely important to be part of the panel proposed26,27,28,29. However, it is established that the high GC content (approximately 80%) in the promoter region of the gene imposes difficulties in the design of an efficient and specific PCR assay to detect and quantify mutations in this region42. This was a technical challenge with TERT C228T mutation. The assay chosen in this study was designed by ThermoFisher Scientific company, but even after several approaches, it was not possible to standardize the assay in the Bio-Rad platform. However, the analysis of this mutation in QuantStudio 3D systems achieved reliable results, as demonstrated by our results both with cell lines and plasma samples.
Despite preliminary, our data suggest an association between ctDNA and worse prognosis (HR: 6.38; 95% CI 1.26–32.1; p = 0.025), as previously described11,18,19,21,22,23,41,45,46,47. From patients with detectable ctDNA, 85% of patients were stage IV, whilst the only stage III patient (ID 12) progressed in 50 days. It is in accordance to other studies that show that the disease burden influences the detection of ctDNA17,18. Also, as reported in literature, ctDNA was not detected in patients with exclusive central nervous system metastases, a fact that could be explained by the presence of the blood–brain barrier, which prevents the detection of circulating DNA21.
Our study demonstrates the impact of ctDNA detection in patient prognosis, in which patients with detectable levels have a worse outcome. Further studies should be done to extend and validate these findings. Moreover, a longitudinal study with several samplings will also allow evaluating the variations in ctDNA levels in patients during the treatment to determine the clinical benefit of the therapies.
Materials and methods
Patient recruitment and study design
We selected naïve-treated melanoma patients with advanced clinical stage III and IV, admitted at Melanoma and Sarcoma Department of Barretos Cancer Hospital from March to November of 2018, or had a follow up appointment during this period. Formalin-fixed paraffin-embedded (FFPE) and peripheral blood samples from patients were collected prospectively at baseline. The present study was approved by the Human Research Ethics Committee of the Barretos Cancer Hospital (1465/2017) and all participants gave their informed written consent. All methods were carried out in accordance with relevant guidelines and regulations.
Identification of somatic mutation
Somatic mutation of hotspot regions of BRAF, NRAS, and TERT genes was determined on formalin-fixed paraffin-embedded (FFPE) tissue available from Pathology Department. Detection of mutation in BRAF and NRAS were performed using the TruSight Tumor 15 panel (Illumina, USA) of next generation sequencing (NGS) as standardized in our institution48. For TERT, mutation analysis was performed by Sanger Sequencing28, or dPCR methodology in those cases that DNA quality did not allow a reliable result from sequencing.
Plasma processing and cfDNA isolation
Between 5 and 10 mL of whole blood were collected in a tube containing EDTA and processed within four hours after collection for plasma separation. To separate plasma from buffy coat, centrifugation was done at 3500 rpm for 10 min and then plasma was stored in 1 mL aliquots in − 80 °C freezer until DNA isolation. cfDNA was isolated from 2 mL of the plasma with the commercial QIAamp Circulating Nucleic Acid Kit (Qiagen, Germany), according to the manufacturer’s protocol and eluted in 35 μL of ultrapure water, quantified using the dsDNA High Sensitivity Assay Kit and Qubit 2.0 Fluorometer (ThermoFisher Scientific, USA) and stored at − 20 °C.
Assays design and Digital PCR platforms
All assays are based on the principle of genotyping, with primer pair delimiting the region of interest and there are two fluorescence/color-labeled probes, one detecting the mutated allele (FAM) and the other one, the wild type allele (HEX/VIC). For BRAF and NRAS, commercial Bio-Rad assays were used (ddPCR BRAF V600 Screening Kit 12001037, capable of detecting mutations in V600E/K/R and ddPCR NRAS Q61 Screening Kit 12001006, capable of detecting mutations in Q61K/L/R/H 183A > T/183A > C). For mutation evaluations in TERT promoter region, TaqMan (ThermoFisher Scientific, USA) assays were used for each mutation, including Hs000000092 (ThermoFisher Scientific, USA) commercial assay for TERT C228T mutation and Hs000000093 (ThermoFisher Scientific, USA) commercial assay for C250T mutation.
BRAF, NRAS and TERT C250T mutations were evaluated in QX200 Droplet Digital PCR (ddPCR) system (Bio-Rad Laboratories, USA). For ddPCR reaction mixture, 1× ddPCR Supermix for Probes (No dUTP), 1× of specific assay and a volume from 1 to 9 μL of DNA were mixed, depending on the concentration of the sample. The final volume of 20 μL of the reaction, together with 70 μL of the Droplet Generation Oil for Probes, was submitted to the QX200 Droplet Generator. PCR reactions were performed in C1000 Touch Thermal Cycler with 96-Deep Well Reaction, droplets were read in the QX200 Droplet Reader and analyzed using Quanta Soft software (Bio-Rad Laboratories, USA). The TERT C228T assay was performed on the Quant Studio 3D platform (ThermoFisher Scientific, USA), after a series of optimization that failure in Bio-Rad platform. For the reaction, 1× Quant Studio 3D Digital PCR Master Mix v2, 1× of the specific assay and 1–6.6 μL of DNA were mixed according to the concentration of the sample. The final volume of 14.5 μL was applied directly to the Quant Studio 3D Digital PCR 20 K Chip v2, and PCR reaction occurred in the Pro Flex thermal cycler using the chip-specific adapter. The chip was inserted into Quant Studio 3D for fluorescence reading and then analyzed by Analysis Suite software (ThermoFisher Scientific, USA).
Validation, optimization, and determination of linearity of DNA amplification
To determine assay’s ability to differentiate mutation positive from mutation negative samples, we tested commercial tumor cell lines whose mutational status are known (Table 2). We evaluated optimal amplification conditions for each assay by testing 20 ng of DNA to a range of temperatures above and below the melting temperature in order to choose the one who provides the optimal cluster separation. Next, a serial dilution containing a mutant ratio of 1:10, 1:100, 1:1000, 1:10,000 and 1:100,000 in a background of wild type DNA (n = three independent replicates) were performed to evaluate the performance and analytical sensitivity of the assays. For this experiment, we tested 10 ng, which is the expected yield for testing samples (cfDNA). Sensitivity was defined as the lowest concentration likely to be reliably distinguished from blank samples.
For the evaluation of assay linearity we compared the concentration in copies/reaction to the expected serial dilution with a linear regression.
Determination of limit of blank (LOB) and limit of detection (LOD)
In order to determine the false positive rate for each assay and accurately determine the limit of detection, we analyzed cell-free DNA extracted from plasma donated by 20 healthy volunteers (blank samples). The LOB was calculated as described by Armbruster and Pry31, where: LOB = meanblank + 1.645*SDblank.
After establishing the LOB, we calculated the lowest analyte concentration likely to be reliably distinguished from the background (LOD) as follow: LOD = LOB + 1.645*(SD low concentration sample)31.
Data analysis of patient samples and positive calls
PCR reactions for patient samples were performed using a maximum input of 10 ng of cfDNA. Each run included mutant positive controls, wild type controls, and non-template controls.
For quality analysis, the overall accepted events/reading should be above 10,000 according to manufacturer’s protocol. To determine positive droplets, the threshold was set examining the three controls wells using the 2D plot. Samples were classified as mutated when number of copies/reaction exceeded the LOB calculated for that assay.
The number of mutated DNA copies per reaction was used to calculate copies per mL of plasma using the following equation:
Copies/mL of plasma = C*EV/TV/PV. PV = Volume of plasma used for cfDNA extraction (mL) EV = Volume in which cfDNA was eluted (μL), TV = Volume of cfDNA added to the PCR reaction (μL), C = copies/reaction (data derived from QuantaSoft or Analysis Suite softwares).
Association between patient detection of ctDNA (undetectable/detectable) and patient’s characteristics (gender, age, AJCC stage, histological classification, primary melanoma Breslow and ulceration, site of metastasis, as well as mutational status) were performed using a Fisher’s exact test. Progression free survival (PFS) was calculated as the time from the date of collection/inclusion to the date of the first disease progression (PD) clinically defined or censored with stable disease at the most recent visit. Median PFS was calculated using the Kaplan–Meier method and compared using the log-rank test. A Cox proportional hazards regression analysis was performed to examine association of ctDNA detection with PFS. Statistical analysis was performed using the IBM SPSS Statistics for Windows version 20.0.
All results were considered significant when p value was < 0.05. Clinical, demographic, histopathological, and treatment-related data, as well as disease progression or relapse data were prospectively collected using a questionnaire and medical records and stored in an electronic database (REDCap).
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This research was funded by grants from the Public Ministry of Labor Campinas (Research, Prevention, and Education of Occupational Cancer) and FAPESP (2018/14468-8). A sincere thanks to Jeremy Squire, for his diligent proofreading of this paper and scientific contribution. We would like to thank to Barretos Cancer Hospital Biobank and the Department of Pathology for the technical support.
The authors declare no competing interests.
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Marczynski, G.T., Laus, A.C., dos Reis, M.B. et al. Circulating tumor DNA (ctDNA) detection is associated with shorter progression-free survival in advanced melanoma patients. Sci Rep 10, 18682 (2020). https://doi.org/10.1038/s41598-020-75792-1
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Brandy and Monica have been set to headline another version of VERZUZ along with the pair could address their previous feud.
In 1998, Brandy and Monica collaborated about the summertime hit’The Lady Is Mine’. Telling the story of 2 girls vying for the identical guy, the accompanying film showed them arguing actor Mekhi Phifer (8 Mile,” ER, Soul Food).
Following the song was published, the business and press appeared a real-life feud involving the brothers.
“It wasn’t something I would say was anybody’s fault,” Monica told Entertainment Tonight. “We all added into it after some time it turned out real… It will get slightly annoying for both I and she once it becomes,’No, I really like her . I like her .'”
Currently, the set may fight it out to another instalment of Swizz Beatz and Timbaland’s favorite VERZUZ series. Monica has triumphed that the pair may explore the fallout throughout the conflict.
“We have long removed ourselves by the dramatics of this we’re gont possess a exact in-depth conversation which may potentially be shared with the crowd since there’s a whole lot that happens, the fans don’t have any idea what actually happened, what actually caused the first rust,” she disclosed.
Asked if they’ll do’The Lady Is Mine’ throughout their VERZUZ, Monica stated it is still being determined.
“I will find out what [Brandy] needs to perform when she eventually reaches Atlanta,” Monica clarified. “We will sit down and determine which way we wish to go and we would like to get it done.”
No matter , the singer claims,”We will be certain it’s still enjoyable and everyone is still using a great time since we need people to become at home ”
She reasoned:”I only anticipate what Monday is. It is going to be a party.”
Brandy and Monica’s VERZUZ occurs on September 1 in 1am BST on Instagram along with Apple Music.
Before this month ), Monica predicted on Kim Kardashian to assist free rapper C-Murder, who had been detained in 2009 following a contentious murder conviction.
Monica, that had been dating the rapper in the time of his sentencing,” clarified that she originally contacted Kardashian — that completed her first year of law school at the beginning of 2020 — in the expectation of procuring her intervention in the instance.
Meanwhile, at a three-star overview of Brandy’s new record’B7′,” NME explained:”It could have taken nearly a long time to get there, however, Brandy’s new album was worth the wait — not merely due to its ear-candy attributes, but also since it’s helped among R&B’s most admired talents locate her grin.”