A permanent cure for HIV infection remains elusive due to the virus's ability to hide away in latent reservoirs. But now, in new research published in print May 3 in the journal Molecular Therapy, scientists at the Lewis Katz School of Medicine at Temple University (LKSOM) and the University of Pittsburgh show that they can excise HIV DNA from the genomes of living animals to eliminate further infection. They are the first to perform the feat in three different animal models, including a "humanized" model in which mice were transplanted with human immune cells and infected with the virus.
The team is the first to demonstrate that HIV-1 replication can be completely shut down and the virus eliminated from infected cells in animals with a powerful gene editing technology known as CRISPR/Cas9. The work was led by Wenhui Hu, MD, PhD, currently Associate Professor in the Center for Metabolic Disease Research and the Department of Pathology (previously in the Department of Neuroscience) at LKSOM; Kamel Khalili, PhD, Laura H. Carnell Professor and Chair of the Department of Neuroscience, Director of the Center for Neurovirology, and Director of the Comprehensive NeuroAIDS Center at LKSOM; and Won-Bin Young, PhD. Dr. Young was Assistant Professor in the Department of Radiology at the University of Pittsburgh School of Medicine at the time of the research. Dr. Young recently joined LKSOM.
The new work builds on a previous proof-of-concept study that the team published in 2016, in which they used transgenic rat and mouse models with HIV-1 DNA incorporated into the genome of every tissue of the animals' bodies. They demonstrated that their strategy could delete the targeted fragments of HIV-1 from the genome in most tissues in the experimental animals.
"Our new study is more comprehensive," Dr. Hu said. "We confirmed the data from our previous work and have improved the efficiency of our gene editing strategy. We also show that the strategy is effective in two additional mouse models, one representing acute infection in mouse cells and the other representing chronic, or latent, infection in human cells."
In the new study, the team genetically inactivated HIV-1 in transgenic mice, reducing the RNA expression of viral genes by roughly 60 to 95 percent, confirming their earlier findings. They then tested their system in mice acutely infected with EcoHIV, the mouse equivalent of human HIV-1.
"During acute infection, HIV actively replicates," Dr. Khalili explained. "With EcoHIV mice, we were able to investigate the ability of the CRISPR/Cas9 strategy to block viral replication and potentially prevent systemic infection." The excision efficiency of their strategy reached 96 percent in EcoHIV mice, providing the first evidence for HIV-1 eradication by prophylactic treatment with a CRISPR/Cas9 system.
In the third animal model, latent HIV-1 infection was recapitulated in humanized mice engrafted with human immune cells, including T cells, followed by HIV-1 infection. "These animals carry latent HIV in the genomes of human T cells, where the virus can escape detection," Dr. Hu explained. Following a single treatment with CRISPR/Cas9, viral fragments were successfully excised from latently infected human cells embedded in mouse tissues and organs.
In all three animal models, the researchers utilized a recombinant adeno-associated viral (rAAV) vector delivery system based on a subtype known as AAV-DJ/8. "The AAV-DJ/8 subtype combines multiple serotypes, giving us a broader range of cell targets for the delivery of our CRISPR/Cas9 system," Dr. Hu said. They also re-engineered their previous gene editing apparatus to now carry a set of four guide RNAs, all designed to efficiently excise integrated HIV-1 DNA from the host cell genome and avoid potential HIV-1 mutational escape.
To determine the success of the strategy, the team measured levels of HIV-1 RNA and used a novel live bioluminescence imaging system. "The imaging system, developed by Dr. Young while at the University of Pittsburgh, pinpoints the spatial and temporal location of HIV-1-infected cells in the body, allowing us to observe HIV-1 replication in real-time and to essentially see HIV-1 reservoirs in latently infected cells and tissues," Dr. Khalili explained.
The new study marks another major step forward in the pursuit of a permanent cure for HIV infection. "The next stage would be to repeat the study in primates, a more suitable animal model where HIV infection induces disease, in order to further demonstrate elimination of HIV-1 DNA in latently infected T cells and other sanctuary sites for HIV-1, including brain cells," Dr. Khalili said. "Our eventual goal is a clinical trial in human patients."
Researchers have developed a much needed alternative to bone grafts that could help alleviate the long-term hospitalization, disability, and considerable costs to the health system associated with non-healing fractures.
Roughly 100,000 broken bones every year in the United States fail to heal properly, resulting in nonunion fractures, and more than 2 million bone grafts are performed around the world annually in attempts to treat these challenging injuries.
Harvesting fresh bone from patients, however, is often painful and donated grafts from tissue banks frequently fail to integrate. Now, Maxim Bez and colleagues devised a two-step gene therapy method coupled with FDA-approved ultrasound and microbubbles that completely healed nonunion fractures in pigs within eight weeks of treatment.
First, researchers placed a collagen scaffold at the site of the break to provide a welcoming niche for bone progenitor cells. Next, they injected microbubbles mixed with genetic material for a bone growth factor. Pulses of sound from an ultrasound wand promoted uptake of the growth factor DNA by progenitor cells, which stimulated bone growth.
Unlike other gene therapies that rely on viral vectors to deliver their cargo -- risky because viruses can permanently integrate into the genome and later promote cancer or set off lethal immune responses -- the ultrasound and microbubbles didn't appreciably trigger inflammation, and expression of the introduced gene was undetectable after 10 days.
The technique was proven to be minimally invasive, safe, and promoted total bone healing, with comparable strength to gold-standard graft procedures.
Bez et al. say that with further development, their system has the potential to be used in many different tissue engineering applications.
Materials provided by American Association for the Advancement of Science. Note: Content may be edited for style and length.
It sounds like the sinking ship that is Donald Trump's administration is about to sink a little harder, and a little faster.
A new report Sunday morning in Axios holds that The Donald is "frustrated and angry at everyone" and is considering a "huge reboot" with his administration.
In other words, he's apparently strongly considering the possibility of firing Sean Spicer, Reince Priebus, and even Steve Bannon.
The report, which cites some of Trump's long time confidants and old friends in which he confides "after dark," holds that the President is being goaded into getting rid of much of his staff, and perhaps even some Cabinet members.
One confidant said about the advice Trump is getting right now:
"The advice he's getting is to go big — that he has nothing to lose. The question now is how big and how bold. I'm not sure he knows the answer to that yet."
Go big and go bold…
So… the advice being given to the man who just fired the FBI Director because of an ongoing investigation of the administration is NOW to try to go big and bold?!
Heaven, help us!!!
Apparently, a part of Trump's complaints include the fact that his Cabinet members and staffers are being too self-centered on their praise or — get this — they aren't sufficiently praising the "brilliant diplomat" that our President believes himself to be.
Read the whole report HERE, and if Spicer, Bannon, Priebus, and others are out later this week, well, don't say we didn't warn you.
[Image via NBC.]
In a story that would warm the cockles of your heart, and possibly even bowels (excuse the pun), this year’s Mother’s Day brings to the fore a story that’s indeed one to be shared. In Uttar Pradesh’s Anantapur village, a 90-year-old woman sold five of her goats to collect enough money to construct a toilet for her 102-year-old mother-in-law.
The woman, Chandana, is now being hailed as an ambassador for the central government’s campaign to improve sanitation in rural India. Apparently, she’s sold off five goats to generate capital to build the toilet as no government authority helped her in this cause, says an ANI report.
80-year-old woman gifts a toilet to her 102-year-old mother-in-law by selling six goats in Kanpur's Anantapur (Uttar Pradesh) pic.twitter.com/wSEgsAKAqu
— ANI UP (@ANINewsUP) May 14, 2017
Chandana says that the inspiration to build the toilet came on the day her mother-in-law had gone out for defecation and fell down, resulting in the centenarian to break her leg. The woman’s son, Ram Prakash, told ANI that the village sarpanch and Kanpur district authorities didn’t do anything even when his mother tried seeking help from them. It was then that she decided to build the toilet by herself.
Tired of having her pleas falling on the deaf ears of the government authorities, Chandana decided that she will take things in her own hand. In spite of being financially unstable, she gathered enough courage to take the decision to sell her goats for this special gift for her mother-in-law.
According to the ANI report, when contacted, the village sarpanch tried to shift the blame on the district authorities. He said, “We have time and again given list to the district authorities but till now not even one toilet has been constructed.”
The district officials appreciated Chandana’s determination and they said that they will conduct an enquiry into the matter.
Locals in Indonesia were baffled by the sudden appearance of a gigantic sea creature that has washed up on a beach.
Since it appeared in Maluku earlier this week, people have suggested it could be a giant squid, whale or octopus.
Whatever it is, the 15 metre-long carcass is pretty disgusting and was still bleeding a day after it was found, according to local media.
Asrul Tuanakota, 37, was the first person to spot the unknown creature but initially believed it was a stranded boat.
It is thought to have been there for days before being discovered, and its skin is discoloured.
M Nasrul Latulanit, a coordinator of Indonesia’s Marine and Coastal Resources Management, said samples have been taken from the scene for laboratory testing.
Four-metre fangs discovered in the carcass indicates it could be a jaw from a whale, he said.
The Sarasota County Sheriff’s Office announced on Friday new charges against deputy Carson Lee Plank. They say the 23-year-old helped her former co-worker Frank Bybee cover up the bizarre attempted murder of an elderly woman.
“As the first responding deputy on January 12, Plank was responsible for initiating the investigation into the attempted murder allegations reported by the victim,” the sheriff’s office said. “Plank however, intentionally ignored the victim’s report and physical evidence to cover up the crime.”
She had obtained an important photo from the victim’s apartment, the SCSO claimed. This depicted blood and hair evidence, but Plank never gave it to investigators.
She faces a new felony charge of tampering with physical evidence, and is out on bond as of Friday. Authorities previously arrested her February 9. Plank got charged with providing false information to law enforcement during an investigation, but actually still has a job with the SCSO. They placed her on administrative leave pending the outcome of criminal and internal investigations. She pleaded not guilty to the original charge, citing lack of evidence, and a pre-trial date had reportedly been set for July 13.
The name Frankie Bybee might be familiar. We wrote about the alleged murder plot before.
The sheriff’s office arrested their now-former deputy in January for embezzling funds meant for looking after an elderly woman’s dog, making out checks to him and his children, and attempting to stage her suicide. His method: He allegedly broke into her home, tried to force feed her medication and later filled the abode with carbon monoxide gas from a running car in the garage. More charges were pressed in February.
Bybee will fight charges in a upcoming trial.
[Image of Plank via Sarasota County Sheriff’s Office]
A 3-foot-long giant rabbit died at a United Airlines pet holding facility in Chicago following a flight from London, in another embarrassment for the airline as it struggles with a global backlash this month over a passenger dragged from his seat.
The 10-month-old Continental Giant breed rabbit named Simon, who was tipped to become one of the world's largest rabbits, had appeared to be in good condition upon arrival at the facility at Chicago's O'Hare airport, an airline spokesman said.
Simon was due to be picked up by a celebrity who had bought him. But when a United worker later checked on Simon, he found he had died, spokesman Charles Hobart said.
"We never want that to happen and it's always a sad experience for all involved when an animal passes while in our care," Hobart said by telephone on Wednesday.
The cause of death has not yet been determined, the spokesman said, adding that United was reviewing what happened.
Hobart said the airline had offered to carry out a post-mortem investigation on the rabbit, but the owner had declined. He said United also offered compensation to the owner, whom he did not identify, but did not disclose the amount.
The incident took place on April 20, but was first reported on Wednesday by The Sun newspaper. Simon's breeder, Annette Edwards, told the paper she was suspicious.
"Simon had a vet's check-up three hours before the flight and was fit as a fiddle," Edwards told The Sun. "Something very strange has happened and I want to know what."
Edwards, a former Playboy model, said she has shipped rabbits all around the world and that nothing like this had ever happened. "The client who bought Simon is very famous. He's upset," she said.
Earlier this month, a United passenger, Dr. David Dao, was unceremoniously dragged from his seat off a plane at O'Hare bound for Louisville, Kentucky, to make room for crew members.
Video recorded by other passengers showed the 69-year-old doctor being dragged down the aisle with blood on his face after he refused to give up his seat on the April 9 flight.
The Mario and Rabbids crossover RPG for Switch first rumored by Laura Kate Dale last November is real, according to Kotaku sources. It is reportedly titled Mario + Rabbids Kingdom Battle and will launch in August or September.
Kotaku got hold of assets from the game that they were asked not to share. The assets allegedly reveal the game is developed on Ubisoft’s Snowdrop engine, the same engine that powered The Division, and that it features turn-based combat, two-player local co-op, and “a goofy sense of humor,” as Kotaku notes the artwork features Mario and friends wielding guns that shoot laser beams. There will apparently be eight playable characters: Mario, Luigi, Yoshi, Peach, and four Rabbids dressed up as Mario, Luigi, Yoshi, and Peach.
Given its supposed August / September release window, it is likely Mario + Rabbids Kingdom Battle will appear at E3 2017 in June, or perhaps even before then.
The Leftovers, this week, once again did that wonderful Leftovers thing of shifting perspectives and taking us into the journey of an important side character for an entire episode.
Scott Glenn's Kevin Garvey Sr. took the honors this time around as we caught up with him during his multi-year trek through the Australian Outback, trying to complete the sacred songline of the indigenous peoples via multiple counts of trespassing and cultural thievery.
"Crazy Whitefella Thinking" was a great episode, though Kevin Sr. is a strange cat to spend this much time with. The bulk of this chapter was played for comedy, or tragicomedy, as Kevin's misadventures in Oz, more often than not, took on a "crazy old fool" quality. Even Kevin accidentally killing the last piece of his song puzzle -- a one Christopher Sunday -- was played for minor laughs. This odyssey could have played out like one of Matt's famous (and splendidly depressing) episode-long ordeals, but instead it had a lightness to it. A lightness that represents The Leftovers' evolution into a series that can juggle and blend many elements and tones.
That's not to say there weren't powerful moments throughout. Kevin Sr. listening to the tape that Kevin Jr. recorded soon after his mother died, where he'd pretended to be a local news reporter, was very touching, as was Kevin Sr.'s affection for it. Which, granted, was also tied to his need for it as a guide on his journey to stop, supposedly, a Biblical flood. Even when the tape was destroyed, you wondered how much he was mourning the loss of a treasured family memory and how much he was grieving over, possibly, the loss of his purpose.
Because make no mistake about it, Kevin Sr. was out for glory. This is HIS story and he did not take kindly to the fact that Matt's been writing about his son being the savior. Kevin Sr.'s the one who's out to rescue us all from drowning in the rising waters of God's wrath. And in the end, after we heard the terrible tale of Grace (Lindsay Duncan) and her dead children, and how she didn't search for them and save them because she thought they'd departed, Kevin Sr. hit us with that silly slap in the face. Had Grace been spilling her guts and unloading on anyone else, she'd have gotten a more sympathetic ear and someone willing to agree that her Kevin the Cop theory was nuts. But she found the one man who was going to keep steering her toward the crazy.
Naturally though, we as viewers know that she's on to something. We've seen younger Kevin's time in the afterlife and know that Matt's new gospel rings true. Still, this is such a great show that just watching someone hit the right note, or find the right path, can make you cringe. Yes, it probably is possible that she can communicate with her dead children, but it still feels like the wrong decision. And Kevin Sr. feels like an insane enabler. But truth be told we're probably watching two of the only people in the world how have an accurate handle on things.
In her sad story too, Kevin Sr. found rediscovered sign post. His eyes almost lit up when he figured out how her story could relate to him. And how he could be the one to help her. Of course, it'll mean getting ahold of his son, who he already resented as someone who might supplant him in this saga, but at that point he just seemed happy get a new sign and welcome someone who, after all this time, would listen to his bats*** ideas.
- While not as supremely surprising and enjoyable as the Perfect Strangers theme last week, Richard Cheese's cover of "Personal Jesus" was a freakin' fun opening credits song. I assume this is to be the trend every week now? A new tune that plays over the Season 2 graphics. I also like how it contrasted with the aboriginal music that scored most of the episode.
- This episode actually reminded me of the premiere's opening movie, about the pious pioneer family who kept standing on their roof and waiting for the End Times, in the sense that Kevin Sr. was so lost, and almost unraveled as a person, that he became desperate to find signs. He was helpless without them. And not just in what we witnessed, with the tape mostly, but his whole time in Australia, which he explained to Sunday. His time taking acid and talking to prophetic chickens. I does make you wonder though why he stopped hearing voices once he left America. Kevin had to go to extraordinary lengths to ditch "passenger" Patti.
- I like how quickly we're getting answers. Like with Nora's arm and baby Lily, the explanations seem to be just a week away. It didn't take long to find out what the end of last week's chapter, with the Kevin drowning, was all about. Remember how we didn't know if that was a flash into the future or what?
- Speaking of answers, what do you think the guy setting himself on fire was about? Do we think it's tied to the scientists who are offering "depart" people using radiation?
- Scott Glenn did a spectacular job this week doing a blend of crotchety and humble. Of course, with the humility of martyrdom also comes that glory-seeking aspect that accompanies being "chosen."
The Leftovers continued its sublime streak of taking things seriously-but-not-too-seriously as we followed Kevin Garvey, Sr.'s desperate vision quest down in Australia. The fact that he's a hero/fool mix, like Don Quixote, works even better because of the dramatic irony at play. We know, for the most part, that he's probably right. Not about every aspect of his erratic journey, but in so far as the voices go. Because we've gone on enough Kevin, Jr. ride-alongs to know the truth. A stirring and fun (connected) detour.
Jamaal Charles is officially a member of the Denver Broncos after agreeing to a one-year deal Tuesday with the AFC West franchise.
The former Kansas City Chiefs star was the franchise's all-time leading rusher with 7,260 yards and 43 touchdowns in nine seasons. He also had 285 receptions for 2,457 yards and 20 scores for the Chiefs.
Charles, 30, is a three-time All-Pro selection and four-time Pro Bowl selection. The former rival running back spent the majority of the last two seasons on injured reserve, playing in just eight games.
The deal is worth up to $3.75 million, according to NFL.com.
"Excited to have Jamaal Charles join the Broncos," Broncos executive vice president of football operations John Elway tweeted. "A great addition to our backfield, and we're thrilled we won't have to play against him!"
Charles tallied at least 1,300 yards from scrimmage in five of six seasons from 2009 through 2014. He had a career-high 1,980 yards from scrimmage and league-best 19 touchdowns in 2013.
The Chiefs cut Charles in February. He tore the ACL in his right leg in 2015 and had meniscus surgery on the same leg in November. He has also had work done on his left knee. Charles visited the Seattle Seahawks in March and was also liked to the Philadelphia Eagles this offseason.
He will fit in somewhere between C.J. Anderson, Devontae Booker, rookie De'Angelo Henderson, Juwan Thompson and Andy Janovich.