Category: hymxgheq

Sangalang gives Star lift but admits he’s still adjusting

first_imgTristan Tamayo/INQUIRER.netIt’s just a matter of time before Ian Sangalang reminds everyone what he’s capable of.And on Sunday, the second overall pick in the 2013 PBA Draft, showed what Star fans have been missing as he led the Hotshots to a 88-77 conquest over the TNT KaTropa.ADVERTISEMENT DILG to lock shops in Tagaytay City, other areas near Taal Filipinos turn Taal Volcano ash, plastic trash into bricks PLAY LIST 01:40Filipinos turn Taal Volcano ash, plastic trash into bricks01:32Taal Volcano watch: Island fissures steaming, lake water receding02:14Carpio hits red carpet treatment for China Coast Guard02:56NCRPO pledges to donate P3.5 million to victims of Taal eruption00:56Heavy rain brings some relief in Australia02:37Calm moments allow Taal folks some respite Ben&Ben, IV of Spades, SB19 win big at 5th Wish Music Awards Panelo: Duterte only wants to emulate strong political will of Marcos Sangalang finished with a team-best 14 points and six rebounds in the Hotshots’ balanced attack to improve their chances of making it to the quarterfinals.And for the Kapampangan big man, he has coach Chito Victolero’s system to thank for his resurgence.FEATURED STORIESSPORTSEnd of his agony? SC rules in favor of Espinosa, orders promoter heirs to pay boxing legendSPORTSBreak new groundSPORTSMcGregor blasts Cerrone in 40 seconds in UFC return“Siguro medyo naga-adjust pa rin ako, pero konti-konti, maa-adapt din namin yung sistema. Ok ako kahit anong mangyari basta nasa game plan,” he said.(Maybe I’m still adjusting, but slowly, we’ll be able to adapt to the system. I’m ok with whatever happens as long as it’s part of our game plan.) Motorcycle taxis ‘illegal’ starting next week — LTFRB board member (We still need to perfect our defense. We can’t take our next games for granted. We need to focus on those because we can’t win those games easily. We really have to work hard for it.)The Hotshots take a momentary breather before facing Blackwater on January 25.Sports Related Videospowered by AdSparcRead Next Bulacan inmates, jail guards raise donations for Taal victims Reverting back to the defensive-minded approach which made the franchise successful, Sangalang echoed Victolero when he said that Star’s chances largely hinge on their defense.“Hindi opensa yung magpapanalo sa amin. Kaya kailangan talaga i-maintain namin yung depensa namin. Yung game plan namin, dapat hindi lalagpas ng 90 yung score nila kaya it’s much better na 77 lang yung naging score nila.”(It’s not our offense that wins us games. We really have to maintain our defense. Our game plan was to limit them below 90 and we did much better in holding them to just 77 points.)As positive as the results from this TNT game may have been, Sangalang acknowledges that the work is just getting started, and Star still has a lot of things to do if it wants to get back to its old championship contending self.“Yung consistency namin sa defense, kailangan pa namin i-perfect,” he said. “Hindi namin pwede i-take for granted yung next games namin. Kailangan mag-focus kami kasi hindi namin makukuha easily yung game. Kailangan namin pagtrabahuhan.”ADVERTISEMENT Palace: Crisis over ABS-CBN franchise unlikely Luis Manzano jokes about Mikee Morada’s proposal to Alex Gonzaga: ‘Baka nagtali lang ng sintas’ MOST READ Marcos monument beside Aquino’s stirs Tarlac town Don’t miss out on the latest news and information. TNT lost effort vs Star, says Castro LATEST STORIES View comments Anne Curtis talks about renewing faith in God amid the world’s ‘noise and clutter’last_img read more

Read More

Reflections after a gap year

first_imgAround this time last year, I wrote about taking time to discover yourself. For high school seniors, August is usually a time for reflection. With examination results coming up, and our illustrious high school careers coming to an end, we must inevitably turn our minds to our next steps.It is definitely a frightening time. Uncertainty and pressure become our closest companions; the former fuelling the latter. Any transition period is daunting, but I feel as though the one wherein we decide what to do after high school is one of the most significant. I feel like this because I think this is one of the first times we take agency over the direction of our lives. Of course, the advice of our parents and trusted elders is important; but, ultimately, the onus falls on us to investigate all possible options and make the best decision for ourselves.Last year, when I was faced with the “What’s next?” question, I chose to take a gap year. It was a difficult decision, but ultimately I think it was the best for me. When you mention taking a gap year to Guyanese, they seem confused by the concept. It’s as though you might as well be giving up on any academic ambition. “So you’re just going to stay home?” Of course not. Gap years have numerous purposes. In my instance, because I knew I wanted to apply to universities in the United States, I took the time to write the SATS (the examinations necessary for entrance to US schools). But even though I knew that what I was doing was for my own good, it doesn’t mean that it wasn’t hard. I had to see my colleagues go off to university or start working in their fields. It felt like everyone was moving forward and I was stuck in a prison of my own design. I’d already written CAPE, at any point, I could have given up and applied to UG, UWI or schools in Canada; but that’s not what I wanted for myself. I knew I wanted to pursue a very specific major available at a few schools, and in the end, I am so happy with how it worked out.Are gap years always beneficial? No. Like everything else, it depends on the person. My gap year really allowed me to explore and develop my passions. With the added free time, I was able to focus more on writing, and I was able to also practice art. Art, for a long time, has been a hobby of mine, but school always took a priority. I have no regrets about that, but I am grateful that I was able to dedicate some time this year to practising art. I was able to meet members of the art community, display my pieces in an art gallery (The Duke), and I was even able to sell three pieces. Knowing that someone likes my art so much that they are willing to pay for it gives me a sense of pride and validation that I don’t think I had previously experienced.I was also able to give back. I love physics, and I think anyone who wants to study the subject should be afforded the best opportunities possible. Unfortunately, physics is somewhat of a neglected science. There are few people who want to do it at higher levels, and even fewer who are willing to teach it. After I had written the SATS, I began to work as a CAPE physics teacher at The Bishop’s High School, and it was both a rewarding and enlightening experience. I developed a newfound respect for teachers, and I was also able to tutor some of the students whom I genuinely consider to be some of the brightest in the country.To current seniors: if you are uncertain about what you want to do, seriously consider a gap year. They can be beneficial, if done right. Don’t let anyone pressure you into making a snap decision; e.g. starting to work at a job you don’t really like just because people keep judging you for “doing nothing”. Don’t just make up your mind on a major just so that you can have an answer when people ask you. Take the time to investigate everything. A year spent in contemplation is nothing compared to a lifetime stuck in a career you hate.last_img read more

Read More

Window of deputy’s car shot out in A.V. Mall lot

first_imgPALMDALE – A gunman shot out a window of a sheriff’s patrol car parked at the Antelope Valley Mall, but no one was hurt, officials said Friday. The deputy was inside the mall at 5:55 p.m. Thursday investigating a shoplifting complaint when someone fired at the car with a small-caliber firearm or possibly a pellet gun, Sgt. Vince Carter said. The car was parked on the north side of the Gottschalks department store in the 1100 block of West Avenue P. Anyone with information about the incident is being asked to call the Palmdale sheriff’s station at (661) 272-2400. The shooting marks the third time Palmdale deputies have been targeted in the past month. On Aug. 3, a sniper fired at a group of a half-dozen deputies involved in a narcotics investigation near an apartment complex at Fifth Street East and Avenue Q. No one was hit. 160Want local news?Sign up for the Localist and stay informed Something went wrong. Please try again.subscribeCongratulations! You’re all set! AD Quality Auto 360p 720p 1080p Top articles1/5READ MOREWhy these photogenic dumplings are popping up in Los AngelesThat incident occurred about a block away from the scene nine days earlier, where a sniper fired twice at a deputy with the Partners Against Crime unit near an apartment building at Fifth Street East and Avenue Q-3. The deputy was getting into his vehicle when he heard a pop and saw a bullet ricochet off the ground. When he turned to look back at the apartments, he heard another shot. The deputy was not hit. karen.maeshiro@dailynews.com (661) 267-5744last_img read more

Read More

Revealed: The improved deal and huge pay rise Spurs will offer Dele Alli

first_imgTottenham Hotspur are confident Dele Alli will sign a new deal, including a ‘significant’ pay rise, at the club before he switches agent.The Telegraph report that Spurs will offer the England international an £80,000-per-week contract – a £30,000 increase on his current salary – and are hopeful his form will notably improve, as a result.Alli is currently searching for a new agent having parted company with Rob Segal, and there is concern that Tottenham could consequently struggle to keep him away from the riches of their rivals, particularly with the club’s strict wage structure.But Spurs are optimistic that an agreement over a new deal will be reached before the 21-year-old formally employs a new representative, which could be crucial in protecting his long-term future in north London. Dele Alli, the Tottenham midfielder 1last_img read more

Read More

Armenia’s manager takes dig at Jose Mourinho over Henrikh Mkhitaryan

first_imgHenrikh Mkhitaryan’s decision to leave Manchester United for Arsenal has been hailed by his international boss.Artur Petrosyan, the Armenia manager, says Jose Mourinho failed to get the best out of the playmaker and he expects him to flourish under Arsene Wenger.The playmaker joined the Gunners on Monday as part of an exchange deal with Alexis Sanchez after 18 months at Old Trafford.“I am annoyed at Mourinho for not playing him,” said Petrosyan. “I am not happy but it is his club, his team, he knows what he wants.“We are very happy with this transfer from Manchester United. His style of play will be better suited at Arsenal.“It is my impression that Henrikh had a problem with the coach and at Arsenal it will not be the same.“There will not be so much pressure on him defensively. Wenger likes his players to express themselves and he likes them to play better football.” 1last_img read more

Read More

Arrangements confirmed for LYIT Donegal’s Presidents’ Cup semi-final

first_imgLYIT Donegal will take to the court for their Hula Hoops Presidents’ Cup semi-final on January 11.Niall McDermott’s team booked a place in the last four with a fine win over Tolka Rovers on Saturday-week past.It is the first time the Donegal side have reached the semi-finals. Basketball Ireland have announced that all semi-final games will take place at Neptune Stadium, Cork and the Parochial Hall, Cork over the weekend of Friday to Sunday, January 10-12th, 2020.  LYIT Donegal face Tradehouse Central Ballincollig in the Parochial Hall at 12 noon on January 11.Arrangements confirmed for LYIT Donegal’s Presidents’ Cup semi-final was last modified: December 10th, 2019 by Chris McNultyShare this:Click to share on Facebook (Opens in new window)Click to share on Twitter (Opens in new window)Click to share on LinkedIn (Opens in new window)Click to share on Reddit (Opens in new window)Click to share on Pocket (Opens in new window)Click to share on Telegram (Opens in new window)Click to share on WhatsApp (Opens in new window)Click to share on Skype (Opens in new window)Click to print (Opens in new window)last_img read more

Read More

Benitez ‘content’ with Newcastle’s January transfer business

first_img Rafael Benitez is in no rush to sign a new contract at Newcastle Rafael Benitez is “content” with Newcastle United’s business in the January transfer window.Benitez was reportedly demanding the club make at least two signings during the window or he would walk away from at the end of the season. Ronaldo warned Lukaku how hard scoring goals in Serie A would be before Inter move Which teams do the best on Boxing Day in the Premier League era? Top nine Premier League free transfers of the decade Where Ancelotti ranks with every Premier League boss for trophies won 3 LATEST FOOTBALL NEWS MONEY ADVICE RANKED huge blow We are delighted to announce the signing of forward Miguel Almirón from @ATLUTD. The 24-year-old has penned a five-and-a-half-year deal which will keep him at St. James’ Park until June 2024.Full story: https://t.co/5NxxCEHhNx #NUFC pic.twitter.com/a5MV5QBTum— Newcastle United FC (@NUFC) January 31, 2019He said: “I didn’t say if we don’t sign players, I will leave. I will not say now that because we have signed two players, I will sign.“We have plenty of time to talk. The most important thing is to concentrate on the next games. My focus was to make sure we had new players.”Whether Ashley and managing director Lee Charnley can clinch Benitez’s signature remains to be seen, but the manager professed himself “happier” at the end of a week which also brought a morale-boosting victory over champions Manchester City. Every time Ally McCoist lost it on air in 2019, including funny XI reactions Oxlade-Chamberlain suffers another setback as Klopp confirms serious injury REVEALED BEST OF The 23-year-old full-back was signed until the end of the season from Ligue 1 side Monaco.Speaking in his press conference ahead of Newcastle’s match at Tottenham Hotspur on Saturday, Benitez said: “Always I use a word that is crucial in life, but especially in football, and that is balance. Rafael Benitez has spoken about Newcastle’s January business Son ban confirmed as Tottenham fail with appeal to overturn red card Almiron signed for Newcastle in a £20million deal The 58-year-old’s squad is largely the same as the one that got promoted from the Championship in 2017 and owner Mike Ashley had been reluctant to splash the cash, as he had hoped to sell the club before the end of the transfer window.Newcastle broke their transfer record on deadline day to sign striker Miguel Almiron from Atlanta United for £20million and also brought in Antonio Barreca on loan. 3 “I am quite pragmatic. I am happier now than one week ago. Why? Because we won against Manchester City and we have brought in two players.”“These two players, they give us something that we didn’t have.“We needed to fill some gaps in terms of positions and in terms of characteristics of players.”The Spaniard also insisted there was “plenty of time” to talk about his future at the club following the end of the transfer window.Benitez’s contract runs out at the end of the season and he has, so far, not agreed a new deal. REVEALED Forbes list reveals how much Mayweather, Ronaldo and Messi earned this decade 3 no dice While Barreca is available to play for the Magpies at Tottenham on Saturday, Almiron will have to wait a little longer as his work permit application is processed.His fee finally broke the record the Magpies set when they signed Michael Owen for £16million in 2005, but Benitez has warned he could take time to settle in English football.He said: “I remember signing a player at Liverpool that people thought he had to work from day one, but I said we had signed him for five years.“We need Almiron right now, but we have not signed him just for this month. He has the desire, ambition and conditions to do well.”last_img read more

Read More

Why Liverpool shouldn’t underestimate tough draw against Villarreal

first_imgA quick search on certain social media platforms is all it takes to get a picture of how Liverpool fans reacted upon being drawn with Villarreal in the semi-finals of the Europa League. Spoiler: while some showed the respect that a team currently placing fourth in La Liga deserves, others were more than a tad dismissive or Marcelino’s side.The latter section may get a nasty shock when the two teams lock horns in the coming fortnight. Let’s be clear: if they defend in the semis the same way they did during large stretches of their quarter-final against Borussia Dortmund, Liverpool will be eliminated.Their slack pressing in the second leg, particularly in the midfield, is exactly the kind of play Villarreal love to lap up. Dortmund’s opener at Anfield came from Liverpool making a mistake while constructing a move, losing the ball high up the pitch and allowing the German side to break quickly (Jürgen Klopp complained after the match about how susceptible his team was to the counter-attack). Right now, there are few teams in Europe better at capitalising upon those sort of situations than the Yellow Submarine. Napoli, one of last year’s Europa League semi-finalists, found that out the hard way in February. So too did Bayer Leverkusen, a good side on their day that has taken positive results from Bayern Munich and FC Barcelona this season. The Reds can’t rely on out-scoring the Spaniards the way they did against Dortmund, either. Real Madrid, Barcelona and Atlético have all dropped points after failing to score more than Villarreal in previous meetings this season. With all respect to Liverpool, those three teams have greater attacking quality than the Premier League’s seventh best.Castellon’s finest are a very, very difficult nut to crack. Deceptively comfortable while riding out periods of opposition dominance of possession, players are well-instructed and aware of the spaces they need to defend, while a firm spirit of sacrifice in the squad helps ensure defensive cover for teammates is swift and effective.Opponents keep knocking on the door, but Villarreal keep absorbing the pressure, until in the blink of an eye, they are at the other end of the pitch with the ball in the net. The second leg of their last Europa League tie with Sparta Prague was a prime example, the Czech supporters at one point letting out “oles” in the stadium, lulled into a false sense of security by the greater share of possession they had at that moment. A few minutes later Samu Castillejo had smashed a shot into the home net, completing a quick move that involved accurate, effective build-up play and a well-timed finish.Denis Suarez in action for Villarreal. John Walton/EMPICS Sport The winger, like Denis Suarez on the opposite flank, is the kind of quick-thinking footballer that excels when given space to work in. If Alberto Moreno in particular gets caught out of position as often as he did in the quarter-finals, they will capitalise on it, and that means opportunities for Cedric Bakambu to score. The striker has found the back of the net six times in his previous four European games, so he isn’t exactly short of confidence.Rather than the path of least resistance to St Jakob-Park as some overly confident supporters seem to think, Liverpool’s next European opponents are perhaps the team remaining in the competition that is best built to take advantage of their weaknesses. Getting past them will likely require a season-best performance for the English side in this competition – certainly one with much more discipline than the Dortmund games. Fail to correct the mistakes from the previous round, and Villarreal, not Liverpool will be on their way to the Europa League final.last_img read more

Read More

Investigation Leads to Meth Arrest of Lanesville Mom

first_imgOn April 1, 2015 troopers from the Indiana State Police Meth Suppression Section assisted officers from the Harrison County Probation Department while conducting a home visit at Berkley’s home in Lanesville.During their investigation troopers obtained a search warrant for the residence and methamphetamine, drug paraphernalia and precursors to manufacture methamphetamine were located.Berkley’s minor children were removed from the residence in accordance with Indiana’s Drug Endangered Children Policy and were placed in the custody of a family member. A Lanesville mother was arrested Wednesday on multiple charges after a search uncovered methamphetamine and meth lab materials at her residence.Amanda Berkley, 33, 7426 Main Street in Lanesville, was arrested on the following charges: Possession of Precursors with the intent to Manufacture – Level 6 Felony, Possession of Methamphetamine – Level 6 Felony, Maintaining a Common Nuisance – Level 6 Felony, Neglect of a Dependent – Level 6 Felony, Possession of Drug Paraphernalia – Class A Misdemeanor.last_img read more

Read More

Mini-antibodies discovered in sharks and camels could lead to drugs for cancer and other diseases

first_img Dooley dispenses 10 milliliters of shark blood into a vial for research studies. Heavy chains Targetmolecule Molecular weight: ~150 kilodaltons (kDa) Typical antibody Small antibody Nanobody binding Nanobody A labmadefragment of acamel or llamasmall antibodythat sharesits binding ability. After anesthetizing a meter-long nurse shark, Dooley carefully lays it out so she can take a blood sample. Miniature antibodies are also starting to prove their worth in patients. Later this year, the first nanobody treatment—derived from a llama small antibody—is expected to receive approval in some countries for use in people with a rare clotting disease. More than 40 similar therapies are in the works for diseases as varied as lupus, lung infections, and cancer. Conventional antibodies are mainstays of diagnosis and staple therapies for a host of diseases, but molecular biologist Nick Devoogdt, also of VUB, predicts that their petite cousins will take over in uses “where conventional antibodies are less optimal.”Small antibodies first came to light by accident, when students at VUB objected to analyzing human blood for a lab exam because they worried about the possibility of contracting a disease. They also refused to kill a mouse to obtain its blood, recalls Serge Muyldermans, a molecular biologist at the university. Some rummaging around, Muyldermans says, turned up an alternative: a stash of frozen dromedary camel serum collected to study the animals’ parasites.But another apparent problem cropped up when the students finished analyzing the blood. Along with normal antibodies, they had sifted out what appeared to be an undersize version of the molecules. “We thought that they had done something wrong,” Muyldermans says. So he and other scientists at the university investigated further. An analysis of blood from zoo animals in the same evolutionary family, including a Bactrian camel and a llama, revealed that all had the same diminutive antibodies.The researchers unveiled their discovery in a 1993 Nature paper and pointed out a key difference between the small antibodies and their full-size counterparts. A conventional antibody consists of four protein strands—two heavy chains that form the backbone of the Y-shaped molecule and two light chains that cling to the outside of the prongs. The mini-antibodies retain roughly the same shape but are missing the light chains. How the unusual molecules might benefit the animals wasn’t clear, but when Dooley learned about them a few years later, she was fascinated. For her Ph.D. research at the University of Aberdeen in the United Kingdom, she set out to investigate those immune oddities. Downsizing antibodies Human blood teems with conventional antibodies—bulky, Y-shaped proteins that home in on bacteria and viruses. The small antibodies produced by sharks and the camel family differ from those immune molecules not only in size, but also in their structure and binding ability. Matt Roth Matt Roth Matt Roth Some researchers are fascinated by unusual immune proteins found in camels and sharks, and this curious nurse shark, surfacing in a tank at the Institute of Marine and Environmental Technology in Baltimore, Maryland, is about to lend a hand—or fin. Like Kobilka and Steyaert, researchers who need a supply of small antibodies or nanobodies often start with llamas, camels, or sharks that have been immunized with particular antigens. In response, the animals’ immune cells begin to make small antibodies that recognize and fasten onto the antigen. Blood cells isolated from the animals allow scientists to obtain the genes for those antibodies, which they can then insert into bacteria or other microorganisms to synthesize large quantities of the immune molecules. But the animals can take a month or more to respond to the antigen and spawn new small antibodies, and llamas and their relatives won’t manufacture versions that target some human proteins that are very similar to their own, notes structural biologist Andrew Kruse of Harvard Medical School in Boston.Earlier this year, he and his colleagues unveiled a synthetic nanobody library that they say overcomes those limitations. Instead of relying on animals as the source of antibody genes, the researchers synthesized DNA sequences to craft their own. Using the published structures and corresponding amino acid sequences of known nanobodies as a guide, they created more than 100 million custom nanobody genes. They slipped the genes into yeast cells, which served as factories for the molecules. Testing showed that “the fully synthetic library is performing at least as well as animal immunization,” Kruse says. He and his colleagues will send it to any academic researchers who pay the shipping costs. Kruse says they are dispatching about 10 to 15 packages a week.Muyldermans and Steyaert, however, contend that animal-derived nanobodies bind to their targets better than entirely labmade alternatives, so they’re sticking with the old-fashioned method. “You never change a winning team, and these llamas are on my team,” Steyaert says.Llamas—along with sharks and camels—may soon be aiding patients. The Ghent, Belgium–based company Ablynx, a spinoff from the original group that discovered the unorthodox antibodies, has already completed a phase III effectiveness trial of one such protein, caplacizumab, for the rare disease acquired thrombotic thrombocytopenic purpura, in which many blood clots can trigger strokes, organ failure, or death. The llama-derived molecule works by latching onto and inactivating a blood protein named von Willebrand factor that promotes clotting—and it’s much more clingy than traditional antibodies, notes Edwin Moses, Ablynx’s CEO. The company, which presented the positive results of the trial in December 2017 at the American Society of Hematology conference in Atlanta, has applied for approval to sell the drug in Europe and plans to do the same in the United States later this year.At least seven other small antibody–derived treatments have reached clinical trials—targeting diseases such as rheumatoid arthritis, psoriasis, and lupus—and more than 30 other treatments are under development. Most of those molecules are derived from the antibodies of camels and their relatives, but the first shark-based drug, produced by the Melbourne, Australia–based firm AdAlta, should enter clinical trials later this year, says Mick Foley, the company’s chief scientific officer. The drug is meant to alleviate lung fibrosis, a stiffening of the organs caused by the buildup of scar tissue. ‹› Typical antibodySed ut perspiciatis unde omnis iste natus error sit voluptatem accusantium doloremque laudantium, totam rem aperiam. Sed ut perspiciatis unde omnis. Helen Dooley admits that she often gets puzzled responses when she describes her work. “People say, ‘You bleed sharks for a living?’”That’s an overstatement, but every couple of weeks she and a helper drop by several large fiberglass tanks at the Institute of Marine and Environmental Technology on the Inner Harbor in Baltimore, Maryland. They net a cat shark or nurse shark and wrestle it into a small pool of water that contains a mild sedative. The drug calms the shark so they can lift it from the water and puncture a vein in its tail. Drawing a few milliliters of blood “doesn’t take more than a few seconds,” Dooley says, after which they return the animal to its aquarium to recover. “They’re usually swimming about perfectly normally, and looking for food, after only a minute or so.”Dooley, an immunologist at the University of Maryland (UMD) School of Medicine in Baltimore, has been tapping shark blood for 2 decades for the same reason that other researchers have been draining the veins of llamas, camels, and their relatives. All those animals pump out unusual, diminutive antibodies that are only about half the size of the conventional versions.Sign up for our daily newsletterGet more great content like this delivered right to you!Country *AfghanistanAland IslandsAlbaniaAlgeriaAndorraAngolaAnguillaAntarcticaAntigua and BarbudaArgentinaArmeniaArubaAustraliaAustriaAzerbaijanBahamasBahrainBangladeshBarbadosBelarusBelgiumBelizeBeninBermudaBhutanBolivia, Plurinational State ofBonaire, Sint Eustatius and SabaBosnia and HerzegovinaBotswanaBouvet IslandBrazilBritish Indian Ocean TerritoryBrunei DarussalamBulgariaBurkina FasoBurundiCambodiaCameroonCanadaCape VerdeCayman IslandsCentral African RepublicChadChileChinaChristmas IslandCocos (Keeling) IslandsColombiaComorosCongoCongo, The Democratic Republic of theCook IslandsCosta RicaCote D’IvoireCroatiaCubaCuraçaoCyprusCzech RepublicDenmarkDjiboutiDominicaDominican RepublicEcuadorEgyptEl SalvadorEquatorial GuineaEritreaEstoniaEthiopiaFalkland Islands (Malvinas)Faroe IslandsFijiFinlandFranceFrench GuianaFrench PolynesiaFrench Southern TerritoriesGabonGambiaGeorgiaGermanyGhanaGibraltarGreeceGreenlandGrenadaGuadeloupeGuatemalaGuernseyGuineaGuinea-BissauGuyanaHaitiHeard Island and Mcdonald IslandsHoly See (Vatican City State)HondurasHong KongHungaryIcelandIndiaIndonesiaIran, Islamic Republic ofIraqIrelandIsle of ManIsraelItalyJamaicaJapanJerseyJordanKazakhstanKenyaKiribatiKorea, Democratic People’s Republic ofKorea, Republic ofKuwaitKyrgyzstanLao People’s Democratic RepublicLatviaLebanonLesothoLiberiaLibyan Arab JamahiriyaLiechtensteinLithuaniaLuxembourgMacaoMacedonia, The Former Yugoslav Republic ofMadagascarMalawiMalaysiaMaldivesMaliMaltaMartiniqueMauritaniaMauritiusMayotteMexicoMoldova, Republic ofMonacoMongoliaMontenegroMontserratMoroccoMozambiqueMyanmarNamibiaNauruNepalNetherlandsNew CaledoniaNew ZealandNicaraguaNigerNigeriaNiueNorfolk IslandNorwayOmanPakistanPalestinianPanamaPapua New GuineaParaguayPeruPhilippinesPitcairnPolandPortugalQatarReunionRomaniaRussian FederationRWANDASaint Barthélemy Saint Helena, Ascension and Tristan da CunhaSaint Kitts and NevisSaint LuciaSaint Martin (French part)Saint Pierre and MiquelonSaint Vincent and the GrenadinesSamoaSan MarinoSao Tome and PrincipeSaudi ArabiaSenegalSerbiaSeychellesSierra LeoneSingaporeSint Maarten (Dutch part)SlovakiaSloveniaSolomon IslandsSomaliaSouth AfricaSouth Georgia and the South Sandwich IslandsSouth SudanSpainSri LankaSudanSurinameSvalbard and Jan MayenSwazilandSwedenSwitzerlandSyrian Arab RepublicTaiwanTajikistanTanzania, United Republic ofThailandTimor-LesteTogoTokelauTongaTrinidad and TobagoTunisiaTurkeyTurkmenistanTurks and Caicos IslandsTuvaluUgandaUkraineUnited Arab EmiratesUnited KingdomUnited StatesUruguayUzbekistanVanuatuVenezuela, Bolivarian Republic ofVietnamVirgin Islands, BritishWallis and FutunaWestern SaharaYemenZambiaZimbabweI also wish to receive emails from AAAS/Science and Science advertisers, including information on products, services and special offers which may include but are not limited to news, careers information & upcoming events.Required fields are included by an asterisk(*)Researchers have known about those tiny proteins since the late 1980s, when scientists at the Free University of Brussels (VUB) stumbled across them. But, “Since 2012, the field has really taken off,” says biochemist Hidde Ploegh of Boston Children’s Hospital. Compared with conventional antibodies, the molecules and even tinier fragments of them, often called nanobodies, are easier for researchers to make, more durable, and more soluble. Small antibodies can work inside cells, and their size allows them to wend deep into tissues, which regular antibodies have a hard time penetrating.Those qualities have established the molecules as valuable research tools. “As a biochemist, I find them exceedingly useful,” Ploegh says. He and his colleagues have deployed them for tasks as diverse as tracking a key immune protein in the body, neutralizing plant viruses, and labeling cancer cells. But the antibodies’ most illuminating research role may be binding to and stabilizing wobbly proteins so that researchers can probe their architecture. “What they’ve been fantastic for is determining crystal structures,” says evolutionary immunologist Martin Flajnik, also of UMD’s School of Medicine. Indeed, a llama nanobody was key to structure work that won a recent Nobel Prize. Matt Roth NanobodySed ut perspiciatis unde omnis iste nataus error sit voluptatem accusantium doloremque accusantium natus error. Nanobody bindingSed ut perspiciatis unde omnis iste natus error sit voluptatem accusantium doloremque accusantium doloremque. That hasn’t stopped scientists from putting them to work. For most medical and research applications, they prune the small antibodies, leaving only the antigen-binding tip. Nanobodies, as they’re called if they come from camels and their relatives, have several handy attributes, Ploegh says. For one, the well-studied bacterium Escherichia coli can make them. “You can express them in E. coli in exceptionally high yield, and they are easy to purify,” he says. In contrast, producing working full-size antibodies in E. coli has proved to be difficult, so researchers typically generate them from more expensive cultures of mammalian cells.Nanobodies also remain functional within cells, whereas conventional antibodies typically fall apart in the cytoplasm. “Suddenly we can do smart things [inside cells] with antibodies,” says developmental biologist Markus Affolter of the University of Basel in Switzerland. One clever use, he says, is “a completely new way to manipulate proteins.”He and his colleagues, for example, harnessed nanobodies to eliminate specific proteins from cells. The team started by engineering the cells to produce a nanobody that, on its nonbinding end, carried molecules that direct proteins to the cell’s garbage disposal. The other end of the nanobody recognized and attached to a specific protein. Stimulating a cell to manufacture the nanobody caused the protein target to disappear in as little as 3 hours, the researchers reported in 2012.Depleting a protein by inducing mutations in its gene, a standard approach in basic research, banishes it for good. In contrast, the nanobody technique allows scientists to delete and then restore the molecule, probing its role in more detail. In 2015, for example, a team led by researchers at Baylor College of Medicine in Houston, Texas, engineered fruit flies to generate a nanobody that grabs the protein Dunce—which is essential for insects’ ability to learn and remember—and promotes its destruction. The researchers then trained the flies to avoid a certain odor. When the team spurred the flies to start making the nanobody against Dunce, levels of the protein plummeted and the insects became dumber—they had a harder time learning to avoid the odor. When the team shut off production of the antibody, Dunce’s abundance bounced back, and the flies became smart again.Because nanobodies reach deep into the nooks and crannies of molecules they bind to, the fragments have also become favorites for researchers trying to stabilize floppy molecules so they can be crystallized, a first step in determining their structure. In perhaps their most impressive performance, nanobodies enabled Brian Kobilka of Stanford’s School of Medicine and his colleagues to capture the first structure of an activated G protein–coupled receptor (GPCR). Those cell membrane proteins relay a multitude of signals into the cell: The adrenaline that makes your heart race, the morphine that eases a patient’s pain after surgery, the bitter flavor in a cup of coffee—all act through GPCRs, which also are the targets of about one-third of drugs. Researchers had tried for about 20 years to get crystal structures of typical, switched-on GPCRs, but the molecules were too shifty.Then Kobilka met structural biologist Jan Steyaert of VUB at a conference, and the two decided to try nanobodies. The researchers and their colleagues injected a llama with the human β2-adrenergic receptor, which responds to adrenaline, and from the animal’s blood prepared a set of nanobodies that clamped onto the receptor. In a 2011 Nature paper, the scientists revealed that one nanobody locked the GPCR into an active shape, allowing them to determine its crystal structure. “That was a devastatingly effective use” of the molecules, Garcia says, as evidenced by the fact that in 2012, Kobilka shared the Nobel Prize in Chemistry. Dooley works quickly to rejuvenate the anesthetized shark once it is back in the water.  She must manually mimic the shark’s swimming movements to generate water flow over the gills. No light chains ~15 kDa Matt Roth Deepbindingpocket Dooley works quickly to rejuvenate the anesthetized shark once it is back in the water.  She must manually mimic the shark’s swimming movements to generate water flow over the gills.center_img Matt Roth By Mitch LeslieMay. 10, 2018 , 12:15 PM Two light and two heavy chains intertwine to make a protein that can identify and affix to bits of pathogens or other molecules. C. BICKEL/SCIENCE Mini-antibodies discovered in sharks and camels could lead to drugs for cancer and other diseases Small antibodies and nanobodiesSed ut perspiciatis unde omnis iste natus error sit voluptatem accusantium doloremque laudantium, totam rem aperiam, eaque ipsa quae ab illo inventore veritatis et quasi architecto beatae vitae dicta sunt explicabo. Nemo enim ipsam voluptatem quia voluptas sit aspernatur aut odit aut fugit. Helen Dooley periodically captures sharks in research aquaria to obtain small antibodies. Matt Roth Some researchers are fascinated by unusual immune proteins found in camels and sharks, and this curious nurse shark, surfacing in a tank at the Institute of Marine and Environmental Technology in Baltimore, Maryland, is about to lend a hand—or fin. The diet of the research sharks is mainly fish, but they get treats of squid and shrimp. Because of its binding style, a nanobody can fit into crevices on molecules. ~90 kDa Light chains She asked VUB researchers for help obtaining samples, and they arranged for colleagues in Morocco to immunize a few camels and send her some blood. After the first shipment arrived, however, the animals mysteriously disappeared, possibly because they were stolen, she says. “I was a year into my Ph.D., and I had no Ph.D. project.” But she soon found an alternative to camels. In 1995, Flajnik and colleagues fished an unconventional antibody out of blood from a nurse shark. Like the camel versions, the shark antibody was smaller than the regular variety and lacked light chains. Dooley contacted Flajnik and finished her Ph.D. research on the shark molecules. Then she continued the work as a postdoc in his lab. “That’s when we started to dig into the nitty-gritty of shark antibodies,” she says.Humans and mice occasionally churn out antibodies that contain only heavy chains, but researchers think they are duds produced by malfunctioning B cells, the immune factories for such proteins. In contrast, the undersize antibodies of sharks and the camel family are not half-finished rejects. Even though they lack the light chains that help regular antibodies recognize and grab antigens, they can bind tightly to their targets with great specificity, and they appear to be a key part of the animals’ response to pathogens. When Dooley and Flajnik injected immune-stimulating antigens into nurse sharks, they discovered that within a few months—the shark immune system is slower to react than ours—the animals were churning out a variety of small antibodies that targeted the foreign molecules. “It’s the major antibody used in the nurse shark,” Flajnik says.He and other researchers speculate that the minute antibodies enable immune systems to counteract a broader range of pathogens. Conventional antibodies excel at sticking to flat surfaces on viral and bacterial molecules. Heavy-chain–only antibodies “are skinny and might be able to penetrate canyons and crevices that regular antibodies couldn’t get into,” says structural immunologist K. Christopher Garcia of the Stanford University School of Medicine in Palo Alto, California.Small antibodies must have an important role, Muyldermans notes, because they emerged independently in as many as three different lineages: sharks, camels, and probably another group of sharklike fishes. But just what advantage they confer on those animals is a mystery. Scientists know little about shark pathogens, for example, and aren’t even sure how to tell when sharks are sick, Dooley says. “They don’t just curl up in the corner.” Even in camels and their kin, whose diseases are better understood, researchers haven’t identified which pathogens the small antibodies combat. At the moment, Muyldermans says, “We have no clue” why the molecules evolved. Helen Dooley started studying the small antibodies of sharks when she lost access to blood samples of camels, which have similar immune proteins. Small antibodySed ut perspiciatis unde omnis iste natus error sit voluptatem accusan-tium doloremque. Matt Roth Helen Dooley of the University of Maryland School of Medicine in Baltimore feeds the sharks. She studies their immune responses, specifically the small antibodies they produce. This slimmed-down variety lacks light chains but can still bind to its targets. Matt Roth Matt Roth Researchers also hope the unique properties of small antibodies will enable them to pry open the blood-brain barrier, an obstacle to treating many brain diseases because it rebuffs most large molecules, including standard antibodies and many other drugs. Ossianix, a Philadelphia, Pennsylvania–based biotech, has crafted a shard of a shark small antibody that binds to a receptor controlling access across the barrier. By stimulating the receptor, the fragment might open the way for drugs such as rituximab, a cancer-killing conventional antibody, to cross into the brain, says Ossianix CEO Frank Walsh.Other applications beckon. Researchers can equip small antibodies with therapeutic cargoes, such as other drugs or cancer-killing radioactive compounds, without making them so massive that they can’t infiltrate tissues or tumors. In addition, the kidneys rapidly filter the molecules out of the blood for excretion—a benefit when the cargo is radioactive. Diminutive antibodies can also be fused to radioactive or fluorescent tracers to illuminate tumors or even guide surgery. To visualize a tumor, for example, “you need small molecules that bind tightly” to the cancer, Devoogdt says. Rapid clearance of a tracer is a plus, as well, because it reduces background noise that can leave the tumor harder to discern. He and his colleagues, along with several other groups, have performed safety trials on nanobodies as tumor-visualizing agents, and they hope to start testing the molecules’ therapeutic usefulness in patients next year.Small antibodies do have some drawbacks for medicine, researchers caution. The rapid excretion of the molecules can be a downside if they leave the body before patients receive their full benefit. Another shortcoming, notes biochemist Jan Gettemans of Ghent University, is that unlike many important drugs, including statins and the anti-HIV drug azidothymidine, the antibodies can’t enter cells on their own. Researchers can get them into the cytoplasm by genetically altering cells to produce them, but that’s not feasible for most treatments. And although brief studies suggest small antibodies are safe for patients, “there is no clear experience in humans … for a long period of time,” Gettemans says. But he and other researchers think the small antibodies’ advantages outweigh their shortcomings and are confident that they “will be part of the armamentarium,” as Walsh puts it.Meanwhile, the mysteries they pose continue to entrance Dooley. She did a stint in drug development—4 years working on a pharmaceutical company’s small-antibody project—but these days when she takes a tube of shark blood back to her lab, she’s usually trying to answer basic questions such as how the animals switch on their antibody-manufacturing cells. She has become fond of the sharks, too. “They are beautiful animals to work with.” If someone did steal the camels she was originally planning to study, she says, “they really did me a favor.”*Correction, 11 May, 2:10 p.m.: An earlier version of this story misstated Hidde Ploegh’s affiliation.last_img read more

Read More