text:
Serbian Orthodox Cathedral, Zagreb
|Serbian Orthodox Cathedral in Zagreb|
Cathedral of the Transfiguration of the Lord
Zagreb Orthodox Cathedral from Petar Preradović Square
Serbian Orthodox Cathedral in Zagreb
Shown within Croatia
|45°48′46″N 15°58′26″E / 45.81265°N 15.9739°ECoordinates: 45°48′46″N 15°58′26″E / 45.81265°N 15.9739°E|
|Previous denomination||all Eastern Orthodox communities|
|Former name(s)||Church of St. Peter and Paul (in 1794)|
|Dedication||Transfiguration of the Lord|
Metropolitan Emilijan Marinović (1969-1977)
|Heritage designation||Register of Cultural Goods of Croatia|
|Architect(s)||Franjo Klein and Hermann Bollé|
|Style||Historicist interpretation of Romanesque and Byzantine architecture|
|Archdiocese||Metropolitanate of Zagreb and Ljubljana|
Zagreb Orthodox Cathedral or Cathedral of the Transfiguration of the Lord (Serbian Cyrillic: Храм преображења Господњег, Croatian: Hram preobraženja Gospodnjeg) is a Serbian Orthodox Cathedral located on the Petar Preradović Square in Zagreb, Croatia. It was built in 1865–66 according to designs of architect Franjo Klein. It is ecclessiastically part of the Metropolitanate of Zagreb and Ljubljana and its cathedral.
Old St. Marguerite church
A wooden Catholic church dedicated to St. Marguerite was located on the place of the modern day cathedral in the 14th century. The church was restored in the 16th and 17th century. Between 1372 and the 19th century, the annual St. Marguerite fair was organized on the square. In the 18th century the church was burned down in a fire and in its place a new one was built with bulbous steeple.
Duding the Josephinism period the state implemented significant reforms that affected life of religious communities. In 1781 Patent of Toleration extended religious freedom to non-Catholic Christians living in Habsburg lands and was followed by 1782 Edict of Tolerance. By city government decision the old church was offered at auction and sold to the Zagreb Orthodox Parish for 4000 Austro-Hungarian forint. In 1848, during Revolutions of 1848, the Orthodox Parish added the suffix Serbian in its name since by that time the Serbs significantly outnumbered local Greeks and Aromanians.
Construction of new church
In 1861 initiative was launched to build new church on the site of a dilapidated old St. Marguerite church. Project was awarded to the architect Franjo Klein. In the same period when the Serbian Orthodox Cathedral was built, Zagreb Synagogue was also built according to the Franjo Klein project. Church was completed on 21 October 1866, and synagogue on 27 September 1867.
In 1897, after completion of urbanization of square south to the church, architect Hermann Bollé proposed plan of monumental reorganization of church. This plan was never implemented, but the same architect developed a plan for restoration of bell tower in its modern-day shape in 1899, and in 1913 based on his plan façade was restored.
World War II
During World War II collaborationist Croatian fascist Ustaše regime of Independent State of Croatia seized all property Serbian Orthodox Church and determined that cathedral will be central church of Croatian Orthodox Church what was part of widespread persecution of Serbs.
First church iconostasis was placed in front of the altar in 1795. This iconostasis was donated to Church of St. George in Varaždin in 1884 when new current iconostasis was built. Orthodox Cathedral iconostasis comprises a total of 34 icons and 4 free-standing walnut pillars.
In Serbian Orthodox Cathedral in Zagreb Metropolitan Jovan Pavlović was buried in 2014 after he held position in 1982-2014 period.
- church at night
- Zagreb Cathedral
- History of Zagreb
- Orthodoxy in Croatia
- Serbs of Croatia
- Kantakuzina Katarina Branković Serbian Orthodox Secondary School
- Pravoslavna crkva na preradovićevom trgu, PhD Dragan Damjanović, Zagreb-moj grad, pages 11-13, Issue 28, year IV, May 2010
- Preradovićev (Cvjetni) trg-ogledalo urbaniteta, PhD Snježana Knežević, Zagreb-moj grad, pages 4-9, Issue 28, year IV, May 2010
- Blic. "U Sabornom hramu u Zagrebu sahranjen mitroplit Jovan" (in Serbian). Retrieved 2 May 2015.
- Преображени храм (Живопис храма Св. Преображења Господњег у Загребу)-Храм преображенный (Роснись храма св. Преображенуя Господня б Загребе)-Transfigured church (Fresgues of the church of St. Transfiguration of the Lord in Zagreb), Irina Buseva Davidova and Dragan Damjanović, Zagreb 2008, trilingual publication in Serbian, Russian and English, NSK CIP 673785
|Wikimedia Commons has media related to Cathedral of the Transfiguration of the Lord in Zagreb.|
Media related to Cathedral of the Transfiguration of the Lord in Zagreb at Wikimedia Commons
- Povijest Trga Petra Preradovića: Iz 19. u 21. stoljeće? (Croatian)
- Arhitektura ikonostasa u opusu Hermana Bolléa, Prostor, Znanstveni časopis za arhitekturu i urbanizam br. 18 (2010), 1 (39); Zagreb, 2010., str. 62 – 79.

domain: geography


It was only last week that Channing Tatum was sharing his love for wife Jenna Dewan Tatum on her 36th birthday, and Jenna, herself, responded with adoration for her husband and Step Up co-star. Now, Jenna is opening up in a frank interview, describing what the experience of falling in love with Channing was for her and also sharing details about their lovemaking, which Ms. Dewan Tatum describes as “primal” at one point. While Jenna and Mr. Tatum have long been adored by a growing mass of followers, the couple has kept their most intimate moments private, until now.
Jenna Dewan Tatum On Falling In Love With Channing Tatum
It’s a question often asked of couples and rarely pinpointed as specifically as Jenna has done, but, as Brides reports, Ms. Dewan Tatum reveals the story of falling in love with Channing. She recalls with clarity the occasion that caused her to be won over by Tatum’s charm, leaving her heart vulnerable and ready to accept Channing’s love.
Fans know that Jenna Dewan and Channing Tatum met on the set of Step Up, their now famous 2006 dance drama, but, until now, the details of how that romance blossomed have been kept private. In describing the momentous day, Jenna recalls that, while they were filming some steamy scenes for Step Up, their off-screen relationship was having trouble getting started. In fact, Jenna reveals that Channing might not have ever talked to her, if he hadn’t enjoyed the party a little too much.
“It was two nights of being weird,” Dewan Tatum says. “We hadn’t even kissed. And then he went out partying with a bunch of dancers. His room was right above mine, and he came down drunk with a sombrero on, banging on my door. He was like, ‘I couldn’t stop thinking about you. Let’s do this. I just want to be with you.’ From that point forward, we were together.”
Jenna Spills The Details On Her “Primal, Earthy” Sex With Channing Tatum
While Jenna Dewan Tatum opened up about sex with her husband for her latest interview with Cosmopolitan, she isn’t the only one to have given insight into their most intimate moments. A Facebook Live chat got steamy, when Channing opened up on his own views about sex, telling fans that he sees sex as something animalistic, something that helps human beings connect at the most basic of levels.
“It’s something that enriches your life… It’s one of the clearest ways that we communicate, it boils things down to a very primal level,” Channing said, during that earlier Q&A session.
Now, in her own interview, Jenna echoed those sentiments as she shared her own thoughts on getting intimate with Channing and on the sexual experience in general.
“I’ve always been a very sexual person,” Jenna explains. “We definitely have a very happy and healthy [sex life]. Something about being a dancer connects you to your physical body. It’s primal, earthy, sexual energy by nature. You feel your body in a certain way. Channing is very much the same way. He’s very in tune with that.”
While our society still tends to resist treating sexuality with an open and healthy attitude, it seems Jenna and Channing have embraced their physical needs in a way that enhances the sexual experience for both of them. From Dewan Tatum’s own words, it can certainly be assumed that the steamy love scenes in Step Up are nothing compared to what Tatum and Jenna experience in their private lives.
Jenna Dewan Tatum recently appeared on an episode of The CW comedy, No Tomorrow.
Channing Tatum will be starring in Logan Lucky, along with Katherine Waterston and Adam Driver, which is due to hit theaters on October 13, 2017.
[Featured Image by Alberto E. Rodriguez/Getty Images]


When most of the working world went online almost 18 months ago, a good computer monitor became essential. Scrolling through tabs has become exhausting, so having another screen for video calls and online multitasking can make working life easier. Despite a country reopening, a good monitor with a practical webcam is an investment worth making, especially if you are self-employed or your workplace has switched to a hybrid model.
Why do you need a monitor with a built-in webcam?
Whether the monitor is for work or personal use, a built-in camera eliminates the need to buy another device and add more wires to your setup. Many webcams these days include facial recognition, providing a more secure option for signing in to your computer and other accounts.
What are the best monitors with webcams?
So whether you’re zooming in, gaming, or just want more screen, here are five of the best monitors with webcams to upgrade your home office or gaming setup.
If you purchase an independently rated product or service through a link on our website, we may receive an affiliate commission.
1. Lenovo ThinkVision T27hv-20 27-20 QHD VoIP Monitor
Lenovo monitors offer many additional specifications for their price, and the Model T27hr-20 is no different. At 27 inches, there’s a solid, good-quality amount of screen to boot. It works great as a work-from-home display, and we love that the camera automatically dims when you step away or when it detects someone looking over your shoulder.
Lenovo has also installed an “ON AIR” light, to alert passers-by that you are in a video conference or need a little privacy. And, perhaps most importantly, it comes with speakers preinstalled, so you don’t have to buy any external audio devices.
2. Asus BE24EQK 23.8 ″ Full HD LCD Monitor
.A great budget option, Asus 23.8 inch HD LCD monitor has been specially designed for videoconferencing. The built-in 2-megapixel HD webcam is adjustable and can be tilted ten degrees in either direction and combined with the anti-echo microphone, the monitor delivers crystal-clear video and sound for live streaming and video calling.
This device features a frameless IPS panel that offers 178 degree horizontal and vertical viewing angles, so you can enjoy stunning visuals in all directions. The image quality remains solid, as it offers Full HD resolution of 1920 x 1080. It also comes equipped with a standard 100 x 100 millimeter VESA mount so you can hang it on the wall or on an arm. screen.
3. Philips Brilliance 499P9H 49 ″ monitor
the Philips Brilliance is not so much a monitor as a cinematic experience. With a width of 49 inches and a curved UHD 5K LED surface, the screen is immersive without being overwhelming. The Brilliance can shave its screen in half, functioning effectively as two monitors in one, with seamless transitions between the two. Although it may need some adjustment, for working speed it is more or less unmatched. It also has a resolution of 5120 x 2440, the most impressive on this list.
The webcam exits smoothly from the monitor and features secure facial recognition that can log you into your Windows devices in under two seconds. When you are done with video calls, you can easily put the webcam back into the unit.
4. Dell P2418HZm 23.8 ″ Monitor
Most of the options considered on this list are full-featured, versatile monitors that can do a little bit of everything. And while Dell P2418HZm Video Conferencing Monitor perhaps more niche, he certainly excels in his specialty. Everything about the 23.8-inch monitor is personalized and optimized for the job. The webcam’s infrared scanners automatically recognize your face and turn on the screen without asking.
Voice control technology works smoothly with a noise-canceling microphone and responsive speakers. And, finally, the thin bezels mean you can stack an additional monitor alongside the Dell without tedious delays to switch between screens. For the basics, as a work from home screen, this is the way to go.
5. HP EliteDisplay E243m 23.8 ″ Monitor
HP EliteDisplay E243m Monitor measures 23.8 inches and is one of the best monitors with a webcam. The pop-up camera swivels in different directions and can be lowered when not in use. It also offers facial recognition to ensure safety.
It certainly looks the room: sleek and backed by a solid stand. It also delivers in terms of performance, thanks to a solid 1000: 1 contrast ratio, as well as a Full HD 1080p display, the color and picture quality doesn’t leave much to be desired.


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Chloe x Halle is an American R&B duo composed of sisters Chloe and Halle Bailey.
Chloe and Halle Bailey were born in Atlanta, Georgia, and raised in Los Angeles, California. They first performed as Chloe x Halle by posting covers of pop songs on YouTube. They launched their YouTube channel at the ages of 13 and 11, respectively, with a cover of Beyoncé's “Best Thing I Never Had”. Their rendition of “Pretty Hurts” by Beyoncé went viral and drew the attention of the singer herself and in 2015, she signed the sister duo to her management company, Parkwood Entertainment.
Chloe x Halle's debut EP, Sugar Symphony, was released April 29, 2016. The EP was accompanied by its singles "Drop" and "Fall". Later in 2016, they made cameo appearances in Beyoncé's Lemonade visual album. They later served as Beyoncé's opening act for the European leg of The Formation World Tour.
In the spring of 2017, Chloe x Halle released their critically acclaimed project, The Two of Us, which featured new music mostly written and produced by the duo. The mixtape was featured on Rolling Stone magazine's Best R&B Albums of 2017 list.
In December 2017, Chloe x Halle released the theme song for the TV series Grown-ish, entitled "Grown". They were added to the cast as series regulars, after initially signing on for recurring roles. Their song "The Kids Are Alright" was featured in the series debut.
On March 23, 2018, the duo released their debut album, The Kids Are Alright. Later, the album and Chloe x Halle were nominated for two Grammy Awards, "Best Urban Contemporary Album" and "Best New Artist", respectively.
"Our debut album The Kids Are Alright means so much to us, it's three years of self-discovery, vulnerability, and growth, all put into one album. We've put our hearts and souls into every lyric, every beat, and we cannot wait for the world to hear."
On December 4, 2019, at a special performance at the Fonda Theatre, the duo announced that they’re sophomore album is Titled “The Ungodly Hour.” They also performed 2 new songs from their new album named “Do it,” and “Rest of your life.”
Chloe x Halle write and arrange all of their songs in their home studio. They also play their own instruments.
Halle considers herself a “jazz-head” and has been listening to Billie Holiday since she was five years. She has cited the singer as one of the major influences on her vocals.
Chloe is inspired by Alternative artists such as Grimes and Björk and loves R&B music.
The duo considers their sound as a combination of their music taste.
“My sister and I like to make music that has interesting sounds and nuances."
On February 3, 2019, Chloe x Halle sang "America the Beautiful" at Super Bowl LIII.
In July 2019, Disney announced that Halle Bailey had been cast as Ariel in the upcoming live-action remake of The Little Mermaid, set to be directed by Rob Marshall. The film is to be released sometime in 2021. Halle's casting created a small controversy on social media, with some claiming the character of Ariel, a fictional mermaid, could not be African American. However, the reactions to the diverse casting choice has been mostly positive.
|Title||Album details||Peak chart positions|
|The Kids Are Alright||
|The Two of Us||
|"I Say So"||2017||Non-album single|
|"Grown" (from Grown-ish)||2018||The Kids Are Alright|
|"The Kids Are Alright"|
|"Happy Without Me" (featuring Joey Bada$)|
|"Who Knew" (from Grown-ish)||2019||Non-album single|
|"Thinkin Bout Me" (from Grown-ish)|
|"Bougie Party"||2017||Dear White People|
|"Warrior"||2018||A Wrinkle in Time (Original Motion Picture Soundtrack)|
|"Shine Bright"||Trolls: Original Motion Picture Soundtrack|
|"Wolf at Your Door"||For the Throne|
|"Enchanted"||El Camino: A Breaking Bad Movie|
|"Lonely Road"||2017||Willow Smith||The 1st|
|"Silver Bells"||2019||Dionne Warwick||Dionne Warwick & The Voices of Christmas|
- Beyoncé – The Formation World Tour (2016)
- Andra Day – Cheers to the Fall Tour (2016)
- Jay-Z and Beyoncé – On the Run II Tour (2018)
Awards and nominations
|2017||NAACP Image Awards||Outstanding New Artist||Themselves||Nominated|
|2018||BET Awards||Best Group||Themselves||Nominated|
|BET Her||"The Kids Are Alright"||Nominated|
|MTV Video Music Awards||Best New Artist||Themselves||Nominated|
|Push Artist of the Year||Themselves||Nominated|
|MTV Europe Music Awards||Best Push Artist||Themselves||Nominated|
|Soul Train Music Awads||Album Of the Year||The Kids Are Alright||Nominated|
|2019||Grammy Award||Best New Artist||Themselves||Nominated|
|Best Urban Contemporary Album||The Kids Are Alright||Nominated|
|Berlin Music Video Award||Best Editing||The Kids Are Alright||Won|
|2020||NAACP Image Awards||Outstanding Supporting Actress in a Comedy Series||Grown-ish||Pending|

domain: artistic


Former U.S. First Lady Michelle Obama will soon be able to be seen in a new cooking show on the Netflix streaming service. Jeroen Meus need not worry though: the cooking show is aimed at children.
The ex-first lady is both the star and executive producer of the new Netflix series Waffles + Mochi, which will be available on the streaming platform next month. The series is aimed at a younger audience and is about two dolls, Waffles and Mochi, who are trying to become chefs. Obama plays a supermarket owner who helps them with that.
“ I am so happy to be a part of this hilarious and heartwarming show. And I’m not saying this just because of the flying basket, ”Obama says. ‘I wished that Waffles + Mochi been there when my daughters were growing up because its the kind of show that is fun to watch with the family. It gives parents peace of mind when they know their little ones are learning something too.
Healthy eating habits
Obama further describes the series as an extension of his work as the former first lady to promote healthy lifestyles among children. “We know everyone loves good food and a good story. And that’s exactly what our curious and adventurous heroes offer, whether they explore the possibilities of the potato, the world of canning or the art of cleaning a cracked egg.
With the help of Obama and a flying magic cart as a guide, Wafer and Mochi embarked on a world tour, Netflix said in a statement. On this journey, they discover new ingredients, different cuisines from around the world, restaurants, farms and households around the world. With renowned chefs, home cooks, children and celebrities, they learn how to prepare dishes with everyday ingredients.
Obama Production House
The show was created by Higher Ground Productions, the new production house of Michelle and her husband Barack Obama. In 2018, the Obamas signed a multi-year contract with Netflix for an unknown but arguably high amount to produce films, series and documentaries.
Waffles + Mochi can be seen on Netflix starting March 16 and currently has ten twenty-minute episodes.


The grandfather of the accused gunman in Tuesday's deadly school shooting in Uvalde, Texas, said he would have reported the weapons if he'd known about them.'I cannot be around weapons' »
On Monday's The Lead With Jake Tapper, a portion of Tapper's interview with Trevor Reed and his family aired, in which they voiced displeasure with the actions of Republican Rep. Marjorie Taylor Greene. Reed is the former Marine who returned to the U.S. following a prisoner swap with Russia last month after spending 985 days in a Russian prison on bogus charges. Legislation pushing for Reed's release was delayed last year due to Greene and other House Republicans.
In 2016, the NRA said on its blog that the European country had one of the lowest murder rates in the world while still having millions of privately owned guns and a few hunting weapons that don't even require a permit. But the Swiss have some specific rules and regulations for gun use. Insider took a look at the country's past with guns to see why it has lower rates of gun violence than the US, where gun death rates are now at their highest in more than 20 years, and the leading cause of death for children and adolescents.
Jack Dorsey cofounded Twitter in 2006 and the company made him a billionaire. From fighting armies of bots to quashing rumors about sending his beard hair to rapper Azealia Banks, Twitter founder Jack Dorsey leads an unusual life of luxury. After cofounding Twitter on March 21 2006, he was booted as the company's CEO two years later, but returned in 2015 having set up his second company, Square — which he rebranded as Block in 2021.
Outgoing Philippine President Rodrigo Duterte sharply criticized Russian leader Vladimir Putin for the killings of innocent civilians in Ukraine, saying while the two of them have been tagged as killers, “I kill criminals, I don't kill children and the elderly.” Duterte, who openly calls Putin an idol and a friend, voiced his rebuke for the first time over Russia's invasion of Ukraine in remarks aired Tuesday where he blamed the three-month old war for the spike in global oil prices that has battered many countries, including the Philippines.
CNN anchor Alisyn Camerota on Wednesday confronted a Texas Republican lawmaker over inaction on gun violence, directly asking him why the state's legislature seemingly puts more vigor toward stripping women's access to abortion than it does in protecting schoolchildren. “What we want to know is what your solution is,” Camerota asked state Rep. James White while standing in front of Robb Elementary School in Uvalde, Texas, where 19 students were killed on Tuesday. Camerota was referencing the state's restrictive 2021 law, which bans abortions after six weeks of pregnancy and allows private individuals to file civil lawsuits against those who perform or facilitate an abortion procedure.
The average gallon of gas in the US hit a high of $4.59 Tuesday, about 51% higher than a year ago. Costs and a demand slowdown could dash expectations for a driving season resembling pre-COVID times. Pain at the pump has gotten so bad that demand for gasoline is dropping just as the summer driving season is about to begin.
Russian and Chinese bombers flew joint patrols near Japanese and South Korea air defense zones on Tuesday in a pointed farewell to U.S. President Joe Biden as he concluded a trip to Asia that rankled Beijing. The patrols came hours after Biden angered China by saying he would be willing to respond militarily to defend Taiwan if it came under Chinese attack, and as he discussed responses to Russia's invasion of Ukraine with leaders of the Quad, which groups the United States with Australia, India and Japan. Japan said it scrambled jets after Russian and Chinese warplanes neared its airspace while Tokyo was hosting the Quad leaders.
In her resignation email—which was sent to 1,400 employees and contractors at Shell—she said despite warnings from the U.N. and the International Energy Agency that there was no safe level of new oil and gas extraction, Shell was planning to explore and extract “much more. I can no longer work for a company that ignores all the alarms and dismisses the risks of climate change and ecological collapse,” Dennett said. I want Shell execs and management to look in the mirror and ask themselves if they really believe their vision for more oil and gas extraction secures a safe future for humanity.
When Gail Curley began her job as Marshal of the U.S. Supreme Court less than a year ago, she would have expected to work mostly behind the scenes: overseeing the court's police force and the operations of the marble-columned building where the justices work. Earlier this month, however, Curley was handed a bombshell of an assignment, overseeing an unprecedented breach of Supreme Court secrecy, the leak of a draft opinion and apparent votes in a major abortion case. People who know Curley, 53, described the former Army colonel and military lawyer as possessing the right temperament for a highly charged leak investigation: smart, private, apolitical and unlikely to be intimidated.
His attack came before school shootings had become something schools prepared for with drills and plans—and before they were routinely carried out by other students. In that innocent time, when the Dunblane school principal heard loud bangs from another part of the school, he thought it was building work that he hadn't been told about. After a teacher burst into his office and told him what was happening, he he ran to the gym, where he witnessed “a scene of unimaginable carnage, one's worst nightmare,” he told the inquiry later.
Doctors and medical associations have spoken up about the shooting in Uvalde, Texas. Gun violence is the leading cause of death for children and teens, and it's been declared a public health crisis. The American College of Physicians has called for background checks and safety trainings for gun owners.
Unfounded claims that the gunman was an immigrant living in the U.S. illegally, or transgender, quickly emerged on Twitter, Reddit and other social media platforms. They were accompanied by familiar conspiracy theories suggesting the entire shooting was somehow staged. The claims reflect broader problems with racism and intolerance toward transgender people, and are an effort to blame the shooting on minority groups who already endure higher rates of online harassment and hate crimes, according to disinformation expert Jaime Longoria.
Here's what Fine said on Twitter: “I have news for the embarrassment that claims to be our president — try to take our guns and you'll learn why the Second Amendment was written in the first place. He made this wretched comment after President Biden responded to the horrific massacre at Robb Elementary School in Uvalde, Texas, on Tuesday, in which 19 children and two adults died. The president said, in part: “When we passed the assault weapons ban, mass shootings went down.
Greene called Senate Republican Leader Mitch McConnell one of Congress' “failed leaders” and said to loud applause that she's part of a “majority who even now wants to see Hillary Clinton in jail. Sending me back to Washington will send a message to the bloodsucking establishment: It is we who will set the policy agenda for the next decade and not them,” Greene said. She added: “We're going to start speaking the truth more forcefully and more loudly than ever before.
House Speaker Nancy Pelosi pushed back Tuesday on the decision by San Francisco's conservative Catholic archbishop to deny her Communion over her support of abortion rights, saying she respects that people have opposing views but not when they impose them on others. Pelosi made her comments on MSNBC's “Morning Joe.” In a letter last week to Pelosi, Archbishop Salvatore Cordileone said he would refuse her Communion after she vowed to codify into law the Supreme Court's Roe vs. Wade decision establishing a constitutional right to abortion.
Los Angeles Clippers guard Norman Powell shared a video of a white woman bothering him at the boxing gym, reported TMZ. The situation escalated resulted in Las Vegas police getting involved and charging her with a misdemeanor. Tuesday, Powell began an Instagram live video to explain how he was interrupted while working out at the local gym.
Big snakes have even bigger appetites, but one large, swimming serpent in Indiana was recently found dead with a “huge” fish lodged in its throat, photos show. Amber Conley and Brad Walton were fueling up at Brookville Lake, about 76 miles southeast of downtown Indianapolis, when they spotted the snake, Conley said in a May 22 Facebook post. “While fueling up today, we came across a dead snake that had choked on a huge fish,” Conley said.
Four weeks ago, Germany agreed to send dozens of anti-aircraft tanks to help defend Ukraine from Russia's invasion, part of what it called a turning-point after decades of military restraint. Berlin says it can deliver the first Gepard tanks in July. That's too slow, a Ukrainian parliamentarian said on Tuesday, as Russian forces launched an assault on the country's east.
A cause has been discovered in the mysterious death of a weeks-old wild horse on North Carolina's Outer Banks. The colt, named Charlie, was the first born of the season and “showed no sign of illness, struggle, or distress” prior to being found dead April 14, the nonprofit Corolla Wild Horse Fund reports. Charlie was just three weeks old at the time.
A Chinese car company is gaining attention for its marketing of a new electric vehicle designed for women. The battery-powered Ora Ballet Cat is made by Great Wall Motors, a Baoding-based automobile manufacturer in Heibei Province, and is loaded with features “specially designed for oriental women,” according to the company. When you look up, the intimate LED super-sized makeup mirror is in front of you, and the light source gear specially designed for oriental women illuminates the skin, and the makeup effect is more perfect,” Great Wall Motors' Ora brand wrote.
China wants 10 small Pacific nations to endorse a sweeping agreement covering everything from security to fisheries in what one leader warns is a “game-changing” bid by Beijing to wrest control of the region. A draft of the agreement obtained by The Associated Press shows that China wants to train Pacific police officers, team up on “traditional and non-traditional security" and expand law enforcement cooperation. China also wants to jointly develop a marine plan for fisheries — which would include the Pacific's lucrative tuna catch — increase cooperation on running the region's internet networks, and set up cultural Confucius Institutes and classrooms.
Since 2014, satirical website The Onion has published virtually the same article — each with the same headline, “'No Way To Prevent This,' Says Only Nation Where This Regularly Happens” — 21 times, each one following a major gun massacre in the United States. On Wednesday, The Onion posted all 21 of the articles on its homepage, calling stark attention to the unabated epidemic of mass gun killings in the country. “UVALDE, TX — In the hours following a violent rampage in Texas in which a lone attacker killed at least 21 individuals and injured several others, citizens living in the only country where this kind of mass killing routinely occurs reportedly concluded Tuesday that there was no way to prevent the massacre from taking place,” reads the latest article.
UVALDE, Texas ― In a stunning moment on Wednesday, former Rep. Beto O'Rourke (D-Texas) confronted Texas Gov. Greg Abbott (R) over gun control policy at a press conference where officials were giving updates on the mass shooting at an Uvalde elementary school. Lt. Gov. Dan Patrick (R) told O'Rourke he was “an embarrassment.”
India's Oil Minister Hardeep Singh Puri told the World Economic Forum in Davos on Tuesday that a crude oil price of $110 a barrel was not sustainable, as the world faces an energy price crisis which is contributing to rising global inflation. Oil prices have surged this year, with Brent crude hitting $139 a barrel in March for its highest price since 2008, after Russia's invasion of Ukraine exacerbated supply concerns. O/R] As countries around the world struggle with the impact of inflation on disposable income, India's Commerce Minister Piyush Goyal said on the same WEF panel that food inflation in the South Asian country was at a "manageable level".
“It could give millions a chance to finally buy a house or start their business or help their kids get an education, too.”
“You signed a contract…If you don’t want to deal with the financial pressure of debt, don’t take out the loan.”
“Without action from Biden, Black student debt will hinder his agenda with respect to eliminating racial inequities.”
“Blanket relief could end up routing too much relief to those who do not need it and too little to those who do.”
“Unlike a number of other policy issues, student debt relief doesn’t need to be pushed through a narrowly controlled Congress.”


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For daily news updates and web-exclusive news items, visit the “AggBeat Online” section of our website at www.aggman.com A new transportation bill is being referred to as ‘devolution lite.’ Two years of funding provides a reprieve, but a lack of federal ...
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text:
|Reviews for I Know What I Like|
| skyeryder01 chapter 1 . 8/26
Hot! Would love a sequel to this!
| SnapeLover2005 chapter 1 . 1/4
*fans self* Whew! That was hott! #HAFHPotterhead
| batcat4eternity chapter 1 . 6/10/2015
Never read this pairing before, but this was immaculate hotness! Loved it.
I also loved Dramione/Draco that was thrown in there. I did not miss it at all :D :D :D
| LilyYumikoRockMe chapter 1 . 3/10/2015
| Dirty girl chapter 1 . 1/18/2015
wow i cummed
| partialtopotter chapter 1 . 12/22/2014
Please write more Pansy/Hermione! I don't care if it's another story, I just need more! You're so great at it!
| SlashboyandboyLover4ever4life chapter 1 . 11/7/2014
AMAZING JOB I LOVE IT! I WANT MORE
| ssweetnothings13 chapter 1 . 5/1/2014
Hot, hot, hot! Love it! You are an amazing writer, keep it up!
| MadTrout chapter 1 . 4/20/2014
Great story! Pansy played her role and stayed in character the whole time. Very plausible. I wish this w as an actual story rather than just a oneshot.
| Kali-blue chapter 1 . 12/9/2013
Hot fun. Thanks, Kali
| tainted-tash chapter 1 . 10/28/2013
I'm not normally into Femme stuff, but this was amazing! And who would have thought of a pairing for Hermione and Pansy?
| Claudilla chapter 1 . 5/19/2013
wow so hot! I adore Pansy and you've made this lesbian oh so happy. Though I have to say we girls get off riding one another too and we do enjoy kissing!
| articcat621 chapter 1 . 12/5/2012
OH. i enjoyed this. :) never thought of the two of them together, but me likey. although i'm sure it has to do with your magical ability to make characters work together. also, you are the queen of lemons. legit, fantastic. :)
| I Stood in Neither chapter 1 . 12/3/2012
*Drools* that's all I can say about darling piece of yumminess ;D
| InmyInk chapter 1 . 11/17/2012
Wow, and I always hate to Pansy, that was delicious, but I agree with what think Hermione, Pansy barely begins to know what is to be with a know-it-all bookworm in sexual information, poor Pansy, if Mione exploded, Pansy will be a nuclear bomb between the legs of an expert to know how to do ... all!
Umm ... delightful, of course I say this as Hermione, as a rookie.!
IF you know well, spare experience

domain: entertainment


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The Marlowe Youth Company (11-14yrs)
Regular theatre making workshops allow junior Youth Company members to express themselves, increase confidence and learn new skills in a fun and friendly environment.
In our first two terms of the year – Autumn 2023 and Spring 2024 – our Junior Youth Companies will explore a range of skills introducing them to all-things performance including movement, musical theatre, script work, storytelling, theatre design, technical skills and circus.
Our Summer 2024 term will focus on bringing the group’s new skills together to create their own piece of theatre for a summer festival showcase.
Please note, due to popular demand we have added a second 11-14yrs class. The other takes place Saturdays 1.30pm-3pm.
Autumn Term Thu 14 Sep – Thu 7 Dec 2023
Spring Term Thu 4 Jan – Thu 21 Mar 2024
Summer Term Thu 18 Apr – Thu 11 Jul 2024


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Linderman told (www.mmajunkie.com/radio) that he's vacated his Cage Warriors heavyweight title and wants the Legacy FC titles in the Light heavyweight & Heavyweight divisions.
Linderman is set to face Robert Drysdale at Legacy FC 19 which takes place April 12 at Allen Event Center in Allen, Texas.
"I'm really looking forward to owning both titles at Legacy,"
"With a win over Drysdale, I should have my shot at either the heavyweight or the light-heavyweight title,"
"I know Drysdale is probably working with some of the best guys in Vegas," Linderman said. "He's watched my videos; he knows I like to stand up and bang. I've been switching a lot of things up that most heavyweight and light heavyweights don't do."

domain: sports


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I found Bonsai Kitty to be a truly amazing site. The ideas this site suggested were really amazing to me; I had never considered cats as an art form before. I would like to share with you all three examples I'm quite proud of.
One of my favorite kinds of bonsai is the informal upright, a tree which looks very similar to a "natural" tree, without the twisted and constrained forms that are sometimes used. I attempted this, with excellent results. Maya is about three years old, and at this point, we don't need to keep her wrapped in copper wire to maintain an excellent and expressive form.
That's about two and a half feet long, and she's only about an 8-pound cat. She is a graceful fixture, and I don't know what we would keep on that shelf if we didn't have her.
Many bonsai cats (and trees, as well) are unfortunately too regular; they fill the space allotted to them so perfectly that some elegance is lost. Without asymmetry, you cannot have true beauty. This one has been carefully kept on one side of the sink we grew him in, with only the tail crossing to the other side, representing the forces of nature. Graystoke is a wonderful and affectionate cat, and we never need to water him!
I always thought carved marble kitty bookends were cute, but lacked some of the warm, fuzzy quality that makes a cat such a great pet. This took a long time, let me tell you, but the posture is wonderful, and with his claws out, he can stick on the shelf wherever you need him, if you don't have a full shelf's worth of books.
Of course, not all my experiments worked... here's a few pictures to show what can go wrong with Kitty Bonsai.


CLEVELAND (AP) — The prospects for construction of a multibillion-dollar petrochemical plant in Ohio appear to have dimmed after a U.S. subsidiary of a South Korean company announced it was withdrawing as a partner in the project.
A joint statement issued early Tuesday by Daelim Chemical USA and the U.S. subsidiary of Thailand's PTT Global Chemical cited the impacts of the COVID-19 pandemic and oil price volatility for yet another delay in deciding whether the plant along the Ohio River would be built.
PTT Global Chemical America President and CEO Toasaporn Boonyapipat in a separate statement said the project remains a “top priority” as the company searches for a new equity partner and works toward "a final investment decision.”
The plant would convert — crack — molecules of ethane, a byproduct of natural gas drilling, into raw materials used to manufacture plastic and other products.
PTTGCA announced its partnership with Daelim in March 2018. A PTTGCA spokesman said Tuesday that the company has thus far spent around $200 million on engineering and planning.
JobsOhio, the state’s private nonprofit economic development arm, has contributed just over $70 million, including a $20 million grant in February to be paid directly to Bechtel Corp. to complete site engineering and site preparation at the site in Belmont County.
“JobsOhio continues working closely with PTTGCA to bring the project to a successful investment decision,” JobsOhio spokesperson Matt Englehart said in a statement Tuesday.
The project is seen as a potential economic savior for an Appalachian region that has long struggled economically from the loss of jobs in steel, aluminum and glass manufacturing decades ago. The plant would create thousands of construction jobs and hundreds of permanent positions. The hope among state and local officials is the plant will spur further development and revitalize the region.
Larry Merry, executive director of the Belmont County Port Authority, said he is not concerned about the news of Daelim's withdrawal and looks forward to an announcement late this year or early next year that the plant will be built.
“We've been dealing with PTT all along, and they're still committed and working every day to move the project forward," Merry said.


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Stable isotopes are essential tools in biological mass spectrometry. Historically, 18O-stable isotopes have been extensively used to study the catalytic mechanisms of proteolytic enzymes1-3. With the advent of mass spectrometry-based proteomics, the enzymatically-catalyzed incorporation of 18O-atoms from stable isotopically enriched water has become a popular method to quantitatively compare protein expression levels (reviewed by Fenselau and Yao4, Miyagi and Rao5 and Ye et al.6). 18O-labeling constitutes a simple and low-cost alternative to chemical (e.g. iTRAQ, ICAT) and metabolic (e.g. SILAC) labeling techniques7. Depending on the protease utilized, 18O-labeling can result in the incorporation of up to two 18O-atoms in the C-terminal carboxyl group of the cleavage product3. The labeling reaction can be subdivided into two independent processes, the peptide bond cleavage and the carboxyl oxygen exchange reaction8. In our PALeO (protease-assisted labeling employing 18O-enriched water) adaptation of enzymatic 18O-labeling, we utilized 50% 18O-enriched water to yield distinctive isotope signatures. In combination with high-resolution matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS/MS), the characteristic isotope envelopes can be used to identify cleavage products with a high level of specificity. We previously have used the PALeO-methodology to detect and characterize endogenous proteases9 and monitor proteolytic reactions10-11. Since PALeO encodes the very essence of the proteolytic cleavage reaction, the experimental setup is simple and biochemical enrichment steps of cleavage products can be circumvented. The PALeO-method can easily be extended to (i) time course experiments that monitor the dynamics of proteolytic cleavage reactions and (ii) the analysis of proteolysis in complex biological samples that represent physiological conditions. PALeO-TimeCourse experiments help identifying rate-limiting processing steps and reaction intermediates in complex proteolytic pathway reactions. Furthermore, the PALeO-reaction allows us to identify proteolytic enzymes such as the serine protease trypsin that is capable to rebind its cleavage products and catalyze the incorporation of a second 18O-atom. Such "double-labeling" enzymes can be used for postdigestion 18O-labeling, in which peptides are exclusively labeled by the carboxyl oxygen exchange reaction. Our third strategy extends labeling employing 18O-enriched water beyond enzymes and uses acidic pH conditions to introduce 18O-stable isotope signatures into peptides.
19 Related JoVE Articles!
Demonstration of Proteolytic Activation of the Epithelial Sodium Channel (ENaC) by Combining Current Measurements with Detection of Cleavage Fragments
Institutions: Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU).
The described methods can be used to investigate the effect of proteases on ion channels, receptors, and other plasma membrane proteins heterologously expressed in Xenopus laevis
oocytes. In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant cleavage sites. Proteolytic activation is a characteristic feature of the amiloride-sensitive epithelial sodium channel (ENaC). The final activating step involves cleavage of the channel’s γ-subunit in a critical region potentially targeted by several proteases including chymotrypsin and plasmin. To determine the stimulatory effect of these serine proteases on ENaC, the amiloride-sensitive whole-cell current (ΔIami
) was measured twice in the same oocyte before and after exposure to the protease using the two-electrode voltage-clamp technique. In parallel to the electrophysiological experiments, a biotinylation approach was used to monitor the appearance of γENaC cleavage fragments at the cell surface. Using the methods described, it was demonstrated that the time course of proteolytic activation of ENaC-mediated whole-cell currents correlates with the appearance of a γENaC cleavage product at the cell surface. These results suggest a causal link between channel cleavage and channel activation. Moreover, they confirm the concept that a cleavage event in γENaC is required as a final step in proteolytic channel activation. The methods described here may well be applicable to address similar questions for other types of ion channels or membrane proteins.
Biochemistry, Issue 89, two-electrode voltage-clamp, electrophysiology, biotinylation, Xenopus laevis oocytes, epithelial sodium channel, ENaC, proteases, proteolytic channel activation, ion channel, cleavage sites, cleavage fragments
A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
Institutions: Peter MacCallum Cancer Centre.
The serine protease Granzyme B (GzmB) mediates target cell apoptosis when released by cytotoxic T lymphocytes (CTL) or natural killer (NK) cells. GzmB is the most studied granzyme in humans and mice and therefore, researchers need specific and reliable tools to study its function and role in pathophysiology. This especially necessitates assays that do not recognize proteases such as caspases or other granzymes that are structurally or functionally related. Here, we apply GzmB’s preference for cleavage after aspartic acid residues in a colorimetric assay using the peptide thioester Boc-Ala-Ala-Asp-S-Bzl. GzmB is the only mammalian serine protease capable of cleaving this substrate. The substrate is cleaved with similar efficiency by human, mouse and rat GzmB, a property not shared by other commercially available peptide substrates, even some that are advertised as being suitable for this purpose. This protocol is demonstrated using unfractionated lysates from activated NK cells or CTL and is also suitable for recombinant proteases generated in a variety of prokaryotic and eukaryotic systems, provided the correct controls are used. This assay is a highly specific method to ascertain the potential pro-apoptotic activity of cytotoxic molecules in mammalian lymphocytes, and of their recombinant counterparts expressed by a variety of methodologies.
Chemistry, Issue 93, Granzyme B, serine protease, peptide thioesters, BOC-Ala-Ala-Asp-S-Bzl, colorimetric substrate, hydrolysis, asp-ase activity
Sigma's Non-specific Protease Activity Assay - Casein as a Substrate
Institutions: Sigma Aldrich.
Proteases break peptide bonds. In the lab, it is often necessary to measure and/or compare the activity of proteases. Sigma's non-specific protease activity assay may be used as a standardized procedure to determine the activity of proteases, which is what we do during our quality control procedures. In this assay, casein acts as a substrate. When the protease we are testing digests casein, the amino acid tyrosine is liberated along with other amino acids and peptide fragments. Folin and Ciocalteus Phenol, or Folin's reagent primarily reacts with free tyrosine to produce a blue colored chromophore, which is quantifiable and measured as an absorbance value on the spectrophotometer. The more tyrosine that is released from casein, the more the chromophores are generated and the stronger the activity of the protease. Absorbance values generated by the activity of the protease are compared to a standard curve, which is generated by reacting known quantities of tyrosine with the F-C reagent to correlate changes in absorbance with the amount of tyrosine in micromoles. From the standard curve the activity of protease samples can be determined in terms of Units, which is the amount in micromoles of tyrosine equivalents released from casein per minute.
To view this article in Chinese, click here
biochemistry, Issue 19, protease, casein, quality control assay, folin and ciocalteu's reagent, folin's reagent, colorimetric detection, spectrophotometer, Sigma-Aldrich
The ITS2 Database
Institutions: University of Würzburg, University of Würzburg.
The internal transcribed spacer 2 (ITS2) has been used as a phylogenetic marker for more than two decades. As ITS2 research mainly focused on the very variable ITS2 sequence, it confined this marker to low-level phylogenetics only. However, the combination of the ITS2 sequence and its highly conserved secondary structure improves the phylogenetic resolution1
and allows phylogenetic inference at multiple taxonomic ranks, including species delimitation2-8
The ITS2 Database9
presents an exhaustive dataset of internal transcribed spacer 2 sequences from NCBI GenBank11
. Following an annotation by profile Hidden Markov Models (HMMs), the secondary structure of each sequence is predicted. First, it is tested whether a minimum energy based fold12
(direct fold) results in a correct, four helix conformation. If this is not the case, the structure is predicted by homology modeling13
. In homology modeling, an already known secondary structure is transferred to another ITS2 sequence, whose secondary structure was not able to fold correctly in a direct fold.
The ITS2 Database is not only a database for storage and retrieval of ITS2 sequence-structures. It also provides several tools to process your own ITS2 sequences, including annotation, structural prediction, motif detection and BLAST14
search on the combined sequence-structure information. Moreover, it integrates trimmed versions of 4SALE15,16
for multiple sequence-structure alignment calculation and Neighbor Joining18
tree reconstruction. Together they form a coherent analysis pipeline from an initial set of sequences to a phylogeny based on sequence and secondary structure.
In a nutshell, this workbench simplifies first phylogenetic analyses to only a few mouse-clicks, while additionally providing tools and data for comprehensive large-scale analyses.
Genetics, Issue 61, alignment, internal transcribed spacer 2, molecular systematics, secondary structure, ribosomal RNA, phylogenetic tree, homology modeling, phylogeny
Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells
Institutions: University of Texas Health Science Center at Houston.
Luminescence Resonance Energy Transfer, or LRET, is a powerful technique used to measure distances between two sites in proteins within the distance range of 10-100 Å. By measuring the distances under various ligated conditions, conformational changes of the protein can be easily assessed. With LRET, a lanthanide, most often chelated terbium, is used as the donor fluorophore, affording advantages such as a longer donor-only emission lifetime, the flexibility to use multiple acceptor fluorophores, and the opportunity to detect sensitized acceptor emission as an easy way to measure energy transfer without the risk of also detecting donor-only signal. Here, we describe a method to use LRET on membrane proteins expressed and assayed on the surface of intact mammalian cells. We introduce a protease cleavage site between the LRET fluorophore pair. After obtaining the original LRET signal, cleavage at that site removes the specific LRET signal from the protein of interest allowing us to quantitatively subtract the background signal that remains after cleavage. This method allows for more physiologically relevant measurements to be made without the need for purification of protein.
Bioengineering, Issue 91, LRET, FRET, Luminescence Resonance Energy Transfer, Fluorescence Resonance Energy Transfer, glutamate receptors, acid sensing ion channel, protein conformation, protein dynamics, fluorescence, protein-protein interactions
High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli
Institutions: Aix-Marseille Université, Commissariat à l'énergie atomique et aux énergies alternatives (CEA) Saclay, France.
Escherichia coli (E. coli)
is the most widely used expression system for the production of recombinant proteins for structural and functional studies. However, purifying proteins is sometimes challenging since many proteins are expressed in an insoluble form. When working with difficult or multiple targets it is therefore recommended to use high throughput (HTP) protein expression screening on a small scale (1-4 ml cultures) to quickly identify conditions for soluble expression. To cope with the various structural genomics programs of the lab, a quantitative (within a range of 0.1-100 mg/L culture of recombinant protein) and HTP protein expression screening protocol was implemented and validated on thousands of proteins. The protocols were automated with the use of a liquid handling robot but can also be performed manually without specialized equipment.
Disulfide-rich venom proteins are gaining increasing recognition for their potential as therapeutic drug leads. They can be highly potent and selective, but their complex disulfide bond networks make them challenging to produce. As a member of the FP7 European Venomics project (www.venomics.eu), our challenge is to develop successful production strategies with the aim of producing thousands of novel venom proteins for functional characterization. Aided by the redox properties of disulfide bond isomerase DsbC, we adapted our HTP production pipeline for the expression of oxidized, functional venom peptides in the E. coli
cytoplasm. The protocols are also applicable to the production of diverse disulfide-rich proteins. Here we demonstrate our pipeline applied to the production of animal venom proteins. With the protocols described herein it is likely that soluble disulfide-rich proteins will be obtained in as little as a week. Even from a small scale, there is the potential to use the purified proteins for validating the oxidation state by mass spectrometry, for characterization in pilot studies, or for sensitive micro-assays.
Bioengineering, Issue 89, E. coli, expression, recombinant, high throughput (HTP), purification, auto-induction, immobilized metal affinity chromatography (IMAC), tobacco etch virus protease (TEV) cleavage, disulfide bond isomerase C (DsbC) fusion, disulfide bonds, animal venom proteins/peptides
An Affordable HIV-1 Drug Resistance Monitoring Method for Resource Limited Settings
Institutions: University of KwaZulu-Natal, Durban, South Africa, Jembi Health Systems, University of Amsterdam, Stanford Medical School.
HIV-1 drug resistance has the potential to seriously compromise the effectiveness and impact of antiretroviral therapy (ART). As ART programs in sub-Saharan Africa continue to expand, individuals on ART should be closely monitored for the emergence of drug resistance. Surveillance of transmitted drug resistance to track transmission of viral strains already resistant to ART is also critical. Unfortunately, drug resistance testing is still not readily accessible in resource limited settings, because genotyping is expensive and requires sophisticated laboratory and data management infrastructure. An open access genotypic drug resistance monitoring method to manage individuals and assess transmitted drug resistance is described. The method uses free open source software for the interpretation of drug resistance patterns and the generation of individual patient reports. The genotyping protocol has an amplification rate of greater than 95% for plasma samples with a viral load >1,000 HIV-1 RNA copies/ml. The sensitivity decreases significantly for viral loads <1,000 HIV-1 RNA copies/ml. The method described here was validated against a method of HIV-1 drug resistance testing approved by the United States Food and Drug Administration (FDA), the Viroseq genotyping method. Limitations of the method described here include the fact that it is not automated and that it also failed to amplify the circulating recombinant form CRF02_AG from a validation panel of samples, although it amplified subtypes A and B from the same panel.
Medicine, Issue 85, Biomedical Technology, HIV-1, HIV Infections, Viremia, Nucleic Acids, genetics, antiretroviral therapy, drug resistance, genotyping, affordable
Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)
Institutions: Louisiana State University Health Sciences Center.
Genetic screens conducted using Drosophila melanogaster
(fruit fly) have made numerous milestone discoveries in the advance of biological sciences. However, the use of biochemical screens aimed at extending the knowledge gained from genetic analysis was explored only recently. Here we describe a method to purify the protein complex that associates with any protein of interest from adult fly heads. This method takes advantage of the Drosophila
GAL4/UAS system to express a bait protein fused with a Tandem Affinity Purification (TAP) tag in fly neurons in vivo
, and then implements two rounds of purification using a TAP procedure similar to the one originally established in yeast1
to purify the interacting protein complex. At the end of this procedure, a mixture of multiple protein complexes is obtained whose molecular identities can be determined by mass spectrometry. Validation of the candidate proteins will benefit from the resource and ease of performing loss-of-function studies in flies. Similar approaches can be applied to other fly tissues. We believe that the combination of genetic manipulations and this proteomic approach in the fly model system holds tremendous potential for tackling fundamental problems in the field of neurobiology and beyond.
Biochemistry, Issue 82, Drosophila, GAL4/UAS system, transgenic, Tandem Affinity Purification, protein-protein interaction, proteomics
Quantitative FRET (Förster Resonance Energy Transfer) Analysis for SENP1 Protease Kinetics Determination
Institutions: University of California, Riverside .
Reversible posttranslational modifications of proteins with ubiquitin or ubiquitin-like proteins (Ubls) are widely used to dynamically regulate protein activity and have diverse roles in many biological processes. For example, SUMO covalently modifies a large number or proteins with important roles in many cellular processes, including cell-cycle regulation, cell survival and death, DNA damage response, and stress response 1-5. SENP, as SUMO-specific protease, functions as an endopeptidase in the maturation of SUMO precursors or as an isopeptidase to remove SUMO from its target proteins and refresh the SUMOylation cycle 1,3,6,7
The catalytic efficiency or specificity of an enzyme is best characterized by the ratio of the kinetic constants, kcat
. In several studies, the kinetic parameters of SUMO-SENP pairs have been determined by various methods, including polyacrylamide gel-based western-blot, radioactive-labeled substrate, fluorescent compound or protein labeled substrate 8-13
. However, the polyacrylamide-gel-based techniques, which used the "native" proteins but are laborious and technically demanding, that do not readily lend themselves to detailed quantitative analysis. The obtained kcat
from studies using tetrapeptides or proteins with an ACC (7-amino-4-carbamoylmetylcoumarin) or AMC (7-amino-4-methylcoumarin) fluorophore were either up to two orders of magnitude lower than the natural substrates or cannot clearly differentiate the iso- and endopeptidase activities of SENPs.
Recently, FRET-based protease assays were used to study the deubiquitinating enzymes (DUBs) or SENPs with the FRET pair of cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) 9,10,14,15
. The ratio of acceptor emission to donor emission was used as the quantitative parameter for FRET signal monitor for protease activity determination. However, this method ignored signal cross-contaminations at the acceptor and donor emission wavelengths by acceptor and donor self-fluorescence and thus was not accurate.
We developed a novel highly sensitive and quantitative FRET-based protease assay for determining the kinetic parameters of pre-SUMO1 maturation by SENP1. An engineered FRET pair CyPet and YPet with significantly improved FRET efficiency and fluorescence quantum yield, were used to generate the CyPet-(pre-SUMO1)-YPet substrate 16
. We differentiated and quantified absolute fluorescence signals contributed by the donor and acceptor and FRET at the acceptor and emission wavelengths, respectively. The value of kcat
was obtained as (3.2 ± 0.55) x107
of SENP1 toward pre-SUMO1, which is in agreement with general enzymatic kinetic parameters. Therefore, this methodology is valid and can be used as a general approach to characterize other proteases as well.
Bioengineering, Issue 72, Biochemistry, Molecular Biology, Proteins, Quantitative FRET analysis, QFRET, enzyme kinetics analysis, SENP, SUMO, plasmid, protein expression, protein purification, protease assay, quantitative analysis
Use of the Protease Fluorescent Detection Kit to Determine Protease Activity
Institutions: Sigma Aldrich.
The Protease Fluorescent Detection Kit provides ready-to-use reagents for detecting the presence of protease activity. This simple assay to detect protease activity uses casein labeled with fluorescein isothiocyanate (FITC) as the substrate.
Protease activity results in the cleavage of the FITC-labeled casein substrate into smaller fragments, which do not precipitate under acidic conditions. After incubation of the protease sample and substrate, the reaction is acidified with the addition of trichloroacetic acid (TCA). The mixture is then centrifuged with the undigested substrate forming a pellet and the smaller, acid soluble fragments remaining in solution. The supernatant is neutralized and the fluorescence of the FITC-labeled fragments is measured.
The described kit procedure detects the trypsin protease control at a concentration of approximately 0.5 μg/ml (5 ng of trypsin added to the assay). This sensitivity can be increased with a longer incubation time, up to 24 hours. The assay is performed in microcentrifuge tubes and procedures are provided for fluorescence detection using either cuvettes or multiwell plates.
Basic Protocols, Issue 30, Protease Fluorescent Detection Kit, Protease Detection, serine proteases, cysteine proteases, metallo-proteases, aspartic proteases
Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry
Institutions: University of Heidelberg.
All cellular processes depend on the functionality of proteins. Although the functionality of a given protein is the direct consequence of its unique amino acid sequence, it is only realized by the folding of the polypeptide chain into a single defined three-dimensional arrangement or more commonly into an ensemble of interconverting conformations. Investigating the connection between protein conformation and its function is therefore essential for a complete understanding of how proteins are able to fulfill their great variety of tasks. One possibility to study conformational changes a protein undergoes while progressing through its functional cycle is hydrogen-1
H-exchange in combination with high-resolution mass spectrometry (HX-MS). HX-MS is a versatile and robust method that adds a new dimension to structural information obtained by e.g.
crystallography. It is used to study protein folding and unfolding, binding of small molecule ligands, protein-protein interactions, conformational changes linked to enzyme catalysis, and allostery. In addition, HX-MS is often used when the amount of protein is very limited or crystallization of the protein is not feasible. Here we provide a general protocol for studying protein dynamics with HX-MS and describe as an example how to reveal the interaction interface of two proteins in a complex.
Chemistry, Issue 81, Molecular Chaperones, mass spectrometers, Amino Acids, Peptides, Proteins, Enzymes, Coenzymes, Protein dynamics, conformational changes, allostery, protein folding, secondary structure, mass spectrometry
High-throughput Fluorometric Measurement of Potential Soil Extracellular Enzyme Activities
Institutions: Colorado State University, Oak Ridge National Laboratory, University of Colorado.
Microbes in soils and other environments produce extracellular enzymes to depolymerize and hydrolyze organic macromolecules so that they can be assimilated for energy and nutrients. Measuring soil microbial enzyme activity is crucial in understanding soil ecosystem functional dynamics. The general concept of the fluorescence enzyme assay is that synthetic C-, N-, or P-rich substrates bound with a fluorescent dye are added to soil samples. When intact, the labeled substrates do not fluoresce. Enzyme activity is measured as the increase in fluorescence as the fluorescent dyes are cleaved from their substrates, which allows them to fluoresce. Enzyme measurements can be expressed in units of molarity or activity. To perform this assay, soil slurries are prepared by combining soil with a pH buffer. The pH buffer (typically a 50 mM sodium acetate or 50 mM Tris buffer), is chosen for the buffer's particular acid dissociation constant (pKa) to best match the soil sample pH. The soil slurries are inoculated with a nonlimiting amount of fluorescently labeled (i.e.
C-, N-, or P-rich) substrate. Using soil slurries in the assay serves to minimize limitations on enzyme and substrate diffusion. Therefore, this assay controls for differences in substrate limitation, diffusion rates, and soil pH conditions; thus detecting potential enzyme activity rates as a function of the difference in enzyme concentrations (per sample).
Fluorescence enzyme assays are typically more sensitive than spectrophotometric (i.e.
colorimetric) assays, but can suffer from interference caused by impurities and the instability of many fluorescent compounds when exposed to light; so caution is required when handling fluorescent substrates. Likewise, this method only assesses potential enzyme activities under laboratory conditions when substrates are not limiting. Caution should be used when interpreting the data representing cross-site comparisons with differing temperatures or soil types, as in situ
soil type and temperature can influence enzyme kinetics.
Environmental Sciences, Issue 81, Ecological and Environmental Phenomena, Environment, Biochemistry, Environmental Microbiology, Soil Microbiology, Ecology, Eukaryota, Archaea, Bacteria, Soil extracellular enzyme activities (EEAs), fluorometric enzyme assays, substrate degradation, 4-methylumbelliferone (MUB), 7-amino-4-methylcoumarin (MUC), enzyme temperature kinetics, soil
Protein WISDOM: A Workbench for In silico De novo Design of BioMolecules
Institutions: Princeton University.
The aim of de novo
protein design is to find the amino acid sequences that will fold into a desired 3-dimensional structure with improvements in specific properties, such as binding affinity, agonist or antagonist behavior, or stability, relative to the native sequence. Protein design lies at the center of current advances drug design and discovery. Not only does protein design provide predictions for potentially useful drug targets, but it also enhances our understanding of the protein folding process and protein-protein interactions. Experimental methods such as directed evolution have shown success in protein design. However, such methods are restricted by the limited sequence space that can be searched tractably. In contrast, computational design strategies allow for the screening of a much larger set of sequences covering a wide variety of properties and functionality. We have developed a range of computational de novo
protein design methods capable of tackling several important areas of protein design. These include the design of monomeric proteins for increased stability and complexes for increased binding affinity.
To disseminate these methods for broader use we present Protein WISDOM (https://www.proteinwisdom.org), a tool that provides automated methods for a variety of protein design problems. Structural templates are submitted to initialize the design process. The first stage of design is an optimization sequence selection stage that aims at improving stability through minimization of potential energy in the sequence space. Selected sequences are then run through a fold specificity stage and a binding affinity stage. A rank-ordered list of the sequences for each step of the process, along with relevant designed structures, provides the user with a comprehensive quantitative assessment of the design. Here we provide the details of each design method, as well as several notable experimental successes attained through the use of the methods.
Genetics, Issue 77, Molecular Biology, Bioengineering, Biochemistry, Biomedical Engineering, Chemical Engineering, Computational Biology, Genomics, Proteomics, Protein, Protein Binding, Computational Biology, Drug Design, optimization (mathematics), Amino Acids, Peptides, and Proteins, De novo protein and peptide design, Drug design, In silico sequence selection, Optimization, Fold specificity, Binding affinity, sequencing
Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
Institutions: The Scripps Research Institute, City College of New York.
The 3’ end of mammalian mRNAs is not formed by abrupt termination of transcription by RNA polymerase II (RNPII). Instead, RNPII synthesizes precursor mRNA beyond the end of mature RNAs, and an active process of endonuclease activity is required at a specific site. Cleavage of the precursor RNA normally occurs 10-30 nt downstream from the consensus polyA site (AAUAAA) after the CA dinucleotides. Proteins from the cleavage complex, a multifactorial protein complex of approximately 800 kDa, accomplish this specific nuclease activity. Specific RNA sequences upstream and downstream of the polyA site control the recruitment of the cleavage complex. Immediately after cleavage, pre-mRNAs are polyadenylated by the polyA polymerase (PAP) to produce mature stable RNA messages.
Processing of the 3’ end of an RNA transcript may be studied using cellular nuclear extracts with specific radiolabeled RNA substrates. In sum, a long 32
P-labeled uncleaved precursor RNA is incubated with nuclear extracts in vitro
, and cleavage is assessed by gel electrophoresis and autoradiography. When proper cleavage occurs, a shorter 5’ cleaved product is detected and quantified. Here, we describe the cleavage assay in detail using, as an example, the 3’ end processing of HIV-1 mRNAs.
Infectious Diseases, Issue 87, Cleavage, Polyadenylation, mRNA processing, Nuclear extracts, 3' Processing Complex
Isolation and Quantification of Botulinum Neurotoxin From Complex Matrices Using the BoTest Matrix Assays
Institutions: BioSentinel Inc., Madison, WI.
Accurate detection and quantification of botulinum neurotoxin (BoNT) in complex matrices is required for pharmaceutical, environmental, and food sample testing. Rapid BoNT testing of foodstuffs is needed during outbreak forensics, patient diagnosis, and food safety testing while accurate potency testing is required for BoNT-based drug product manufacturing and patient safety. The widely used mouse bioassay for BoNT testing is highly sensitive but lacks the precision and throughput needed for rapid and routine BoNT testing. Furthermore, the bioassay's use of animals has resulted in calls by drug product regulatory authorities and animal-rights proponents in the US and abroad to replace the mouse bioassay for BoNT testing. Several in vitro
replacement assays have been developed that work well with purified BoNT in simple buffers, but most have not been shown to be applicable to testing in highly complex matrices. Here, a protocol for the detection of BoNT in complex matrices using the BoTest Matrix assays is presented. The assay consists of three parts: The first part involves preparation of the samples for testing, the second part is an immunoprecipitation step using anti-BoNT antibody-coated paramagnetic beads to purify BoNT from the matrix, and the third part quantifies the isolated BoNT's proteolytic activity using a fluorogenic reporter. The protocol is written for high throughput testing in 96-well plates using both liquid and solid matrices and requires about 2 hr of manual preparation with total assay times of 4-26 hr depending on the sample type, toxin load, and desired sensitivity. Data are presented for BoNT/A testing with phosphate-buffered saline, a drug product, culture supernatant, 2% milk, and fresh tomatoes and includes discussion of critical parameters for assay success.
Neuroscience, Issue 85, Botulinum, food testing, detection, quantification, complex matrices, BoTest Matrix, Clostridium, potency testing
Specificity Analysis of Protein Lysine Methyltransferases Using SPOT Peptide Arrays
Institutions: Stuttgart University.
Lysine methylation is an emerging post-translation modification and it has been identified on several histone and non-histone proteins, where it plays crucial roles in cell development and many diseases. Approximately 5,000 lysine methylation sites were identified on different proteins, which are set by few dozens of protein lysine methyltransferases. This suggests that each PKMT methylates multiple proteins, however till now only one or two substrates have been identified for several of these enzymes. To approach this problem, we have introduced peptide array based substrate specificity analyses of PKMTs. Peptide arrays are powerful tools to characterize the specificity of PKMTs because methylation of several substrates with different sequences can be tested on one array. We synthesized peptide arrays on cellulose membrane using an Intavis SPOT synthesizer and analyzed the specificity of various PKMTs. Based on the results, for several of these enzymes, novel substrates could be identified. For example, for NSD1 by employing peptide arrays, we showed that it methylates K44 of H4 instead of the reported H4K20 and in addition H1.5K168 is the highly preferred substrate over the previously known H3K36. Hence, peptide arrays are powerful tools to biochemically characterize the PKMTs.
Biochemistry, Issue 93, Peptide arrays, solid phase peptide synthesis, SPOT synthesis, protein lysine methyltransferases, substrate specificity profile analysis, lysine methylation
A Protocol for Computer-Based Protein Structure and Function Prediction
Institutions: University of Michigan , University of Kansas.
Genome sequencing projects have ciphered millions of protein sequence, which require knowledge of their structure and function to improve the understanding of their biological role. Although experimental methods can provide detailed information for a small fraction of these proteins, computational modeling is needed for the majority of protein molecules which are experimentally uncharacterized. The I-TASSER server is an on-line workbench for high-resolution modeling of protein structure and function. Given a protein sequence, a typical output from the I-TASSER server includes secondary structure prediction, predicted solvent accessibility of each residue, homologous template proteins detected by threading and structure alignments, up to five full-length tertiary structural models, and structure-based functional annotations for enzyme classification, Gene Ontology terms and protein-ligand binding sites. All the predictions are tagged with a confidence score which tells how accurate the predictions are without knowing the experimental data. To facilitate the special requests of end users, the server provides channels to accept user-specified inter-residue distance and contact maps to interactively change the I-TASSER modeling; it also allows users to specify any proteins as template, or to exclude any template proteins during the structure assembly simulations. The structural information could be collected by the users based on experimental evidences or biological insights with the purpose of improving the quality of I-TASSER predictions. The server was evaluated as the best programs for protein structure and function predictions in the recent community-wide CASP experiments. There are currently >20,000 registered scientists from over 100 countries who are using the on-line I-TASSER server.
Biochemistry, Issue 57, On-line server, I-TASSER, protein structure prediction, function prediction
A High-throughput-compatible FRET-based Platform for Identification and Characterization of Botulinum Neurotoxin Light Chain Modulators
Institutions: The Scripps Research Institute, The Scripps Research Institute.
Botulinum neurotoxin (BoNT) is a potent and potentially lethal bacterial toxin that binds to host motor neurons, is internalized into the cell, and cleaves intracellular proteins that are essential for neurotransmitter release. BoNT is comprised of a heavy chain (HC), which mediates host cell binding and internalization, and a light chain (LC), which cleaves intracellular host proteins essential for acetylcholine release. While therapies that inhibit toxin binding/internalization have a small time window of administration, compounds that target intracellular LC activity have a much larger time window of administrations, particularly relevant given the extremely long half-life of the toxin. In recent years, small molecules have been heavily analyzed as potential LC inhibitors based on their increased cellular permeability relative to larger therapeutics (peptides, aptamers, etc.
). Lead identification often involves high-throughput screening (HTS), where large libraries of small molecules are screened based on their ability to modulate therapeutic target function. Here we describe a FRET-based assay with a commercial BoNT/A LC substrate and recombinant LC that can be automated for HTS of potential BoNT inhibitors. Moreover, we describe a manual technique that can be used for follow-up secondary screening, or for comparing the potency of several candidate compounds.
Chemistry, Issue 82, BoNT/A, botulinum neurotoxin, high-throughput screening, FRET, inhibitor, FRET peptide substrate, activator
Institutions: UVP, LLC, Keck Graduate Institute of Applied Life Sciences.
Immunoblotting (western blotting) is a rapid and sensitive assay for the detection and characterization of proteins that works by exploiting the specificity inherent in antigen-antibody recognition. It involves the solubilization and electrophoretic separation of proteins, glycoproteins, or lipopolysaccharides by gel electrophoresis, followed by quantitative transfer and irreversible binding to nitrocellulose, PVDF, or nylon. The immunoblotting technique has been useful in identifying specific antigens recognized by polyclonal or monoclonal antibodies and is highly sensitive (1 ng of antigen can be detected). This unit provides protocols for protein separation, blotting proteins onto membranes, immunoprobing, and visualization using chromogenic or chemiluminescent substrates.
Basic Protocols, Issue 16, Current Protocols Wiley, Immunoblotting, Biochemistry, Western Blotting, chromogenic substrates, chemiluminescent substrates, protein detection.

domain: chemistry


text:
|Your safe-trip checklist|
|Your safe-trip checklist|
Nothing spoils a vacation like unexpected health problems. Being prepared can take some of the sting out of getting sick while you’re away. So before you leave. . .
- Prepare a first-aid kit. Pack travel-size packages of your usual over-the-counter pain reliever, cold and stomach medicine or any other nonprescription drugs you are likely to need; adhesive bandages; antibacterial ointment; and soothing aloe-based lotions.
- Pack any prescription medications you take regularly. Carry a few extra doses in case you happen to be waylaid, and ask your doctor for a copy of your regular prescriptions.
- Check medication labels for warnings that might affect outdoor fun. For example, some medications require that you stay out of the sun.
- If you wear eyeglasses or contact lenses, take along a spare pair as well as a copy of your prescription.
- If you have a chronic illness, ask your doctor for the name of a physician you can call on while you’re at your vacation destination.
Ah…the promise of summer: sunny days and lazy evenings, barbecues and picnics, getaways and good times. To make sure those times really are good, heed the seasonal safety tips on these two pages.
A great tan may look good, but staying pale is definitely better for your health since skin cancer is directly related to sun exposure. Your best defense is to avoid the sun between 10 a.m. and 3 p.m. If that puts a crimp in your summer plans, put your second best defense into action: Apply plenty of sunscreen with an SPF of at least 15; keep your head and body covered in light, loose layers; and wear a wide-brimmed hat and sunglasses.
If you do get a sunburn, apply aloe vera gel right away and take acetaminophen to ease minor discomfort. You’ll probably be a little dehydrated so be sure to drink lots of water. Cool showers, baths or compresses can soothe the burn and prevent it from worsening. Call your healthcare provider if your skin blisters or if you develop chills, fever or nausea.
If you or your child is stung by a bee, wasp or hornet, remove the stinger and venom sac by scraping it with your fingernail or a credit card. Do not squeeze and pull or you’ll release more venom. Wash the site with soap and water and apply an ice pack to stop the swelling. Be alert for signs of allergic reaction, such as wheezing, labored breathing, nausea or vomiting. Go to an emergency room should any of these symptoms develop. (If you or your child is allergic to bees, ask your healthcare provider about prescribing an emergency kit and keep it handy when you are outdoors.) To avoid getting stung in the first place, wear white or khaki-colored long-sleeved shirts and long pants, avoid scented products and shiny jewelry and never swat at a stinging insect.
Don’t leave your common sense at home when you head beach-, lake- or poolside. Make sure a lifeguard is always present and keep a close eye on children in the water. Make sure all family members are wearing life jackets when riding in a boat.
The best way to avoid contact with poison ivy, poison sumac or poison oak is to know what they look like. The distinctive feature of poison ivy is that its smooth, shiny leaves always occur in groups of three. Poison oak is similar; however, it grows in shrubs and its leaves more closely resemble oak leaves. Poison sumac is far less common but far more toxic. It resembles a shrub or a small tree, and its leaves are arranged in groups of seven to 13 pointed leaflets. From late summer to early spring, the plant has clusters of white berries. If you come into contact with any of these plants, wash immediately with soap and water. Rhuligel, an over-the-counter preparation, may help ease itching, and Benadryl, an oral antihistamine, may also bring relief. Note: Wash in chlorine-based detergent any clothing that comes in contact with the plants.
If you’re planning a family hike, find out if the wooded area you’ll be traveling to is Lyme disease country. Deer ticks carrying the virus hang out on plants and leaves close to the ground, so it’s easy to pick one up without realizing it. To avoid doing that, wear long-sleeved shirts and tuck pants legs into knee socks. If you develop a red, bull’s-eye rash after an outing in a wooded area or develop flu-like symptoms, see your doctor immediately. A course of antibiotics within the first 72 hours will stop the progression of Lyme disease, but left untreated it can cause heart, joint or nervous system problems.
A summer road trip is an American tradition, but be careful not to overdo it. Take a five-minute break every hour to stretch your legs. The break will also help you avoid highway fatigue, a kind of hypnosis caused by driving long stretches of flat, empty road. If you know you’ll be driving through desolate territory, keep a cooler with provisions in your trunk and make sure you have basic safety equipment, such as flares, a spare tire and a jack. And, of course, don’t forget to make sure you and your kids buckle up!

domain: medical


text:
Perfect for families who want the peace and quiet of the countryside but with many amenities within a short walk. Spend your days lazing in the garden or relaxing by the fire with good company.
A gracefully converted schoolhouse brimming with artistic touches situated in a leafy corner of Bakewell; this elegant hideaway is ideal for multigenerational families or groups of friends seeking a luxurious retreat in the Peak District. Saddle up and peddle through the serene green countryside across the impressive Headstone Viaduct bridge on the traffic-free Monsal Trail, before stretching out weary feet in the ambient glow of the dual-aspect wood burner.
This beautifully appointed property boasts light-flooded rooms, bold interior styling, and opulent jewel tones throughout. Step through the front door into the contemporary kitchen with sleek handless cabinets and feature tiling, a wonderful space to whip up some post-adventure tipples and snacks. Pull out the wooden bench seat and tuck into a spread of local farm treats or pop a bottle and gather in the dining room for a decadent dinner. Retreat to the lounge and sink onto the plush velvet sofa or slide open the Bifold doors and let the balmy breeze drift inside. For chillier evenings, fire up the wood burner and unwind with a movie or group games. Each of the four bedrooms is thoughtfully styled with soft pillows and throws, consisting of a super-king, two kings, and a twin room. For a moment of peace, sink into the curved copper hot tub and sip on a cold drink – bliss! Head outside and recline on the patio or gear up for garden games.
Saunter into the bustling hub of Bakewell and browse the independent eateries, local shops, galleries, and traditional pubs, less than 500 metres. Trek along the open moorland on the Cork Stone and Nine Ladies Circle trail, 6 miles, or venture to Chatsworth House and stroll the gardens and pick up some treats from the farm shop, 5 miles.

domain: travel


ALBANY, N.Y. (NEWS10) — A Troy man has pled guilty in a theft case, according to the United States Attorney’s Office. Michael McDermott, 30, now faces up to 10 years in prison, a fine of up to $250,000, and a term of supervised release of up to 3 years.
McDermott admitted to breaking into the Glenmont Walmart on May 10 while the business was closed, smashing the glass gun case, and stealing six firearms. He also admitted to pointing a firearm at employees upon them chasing him into the woods.
McDermott was arraigned on May 23 and his trial began at the end of July. His sentencing is scheduled for February 26, 2024.


Chelsea manager, Thomas Tuchel, will have a strong squad to make his selection from when they face Liverpool in the Carabao Cup final on Sunday.
Tuchel is looking to add more silverware to the Blues’ trophy cabinet.
They go into the clash at Wembley following a 2-0 win over Lille in the Champions League round of 16 first leg.
Ahead of the game, Tuchel has delivered a very positive update, stating that everyone has participated in the club’s latest training session, barring Ben Chilwell.
Right-back Reece James therefore could be set to feature after a lengthy spell out on the sidelines, according to Football London.
Goalkeepers: Edouard Mendy, Kepa Arrizabalaga, Marcus Bettinelli.
Defenders: Antonio Rudiger, Malang Sarr, Trevoh Chalobah, Marcos Alonso, Reece James, Cesar Azpilicueta, Thiago Silva, Kenedy.
Midfielders: Mason Mount, N’Golo Kante, Mateo Kovacic, Saul Niguez, Jorginho, Ross Barkley.
Forwards: Kai Havertz, Hakim Ziyech, Christian Pulisic, Callum Hudson-Odoi, Romelu Lukaku, Timo Werner.