Earlier this year, Michigan-based Sintercore released what they called the first commercially-available 3D printed muzzle brake for the AR-15, and now the company has announced the availability of a 3D printed magazine extension for the Glock 43.
The original Glock handgun was created in 1982 by a curtain rod and military knife manufacturer, Gaston Glock, who decided to manufacture the new gun from scratch. After polling a wide range of gun experts in his native Austria, he arrived at what he believed would be the formula to improve the handgun and modernize the design of such weapons.
Glock’s gun would hold more ammunition, be more durable and reliable, easier to fire and simple to learn to operate. And it worked. The original Glock 17, a model adopted by the Austrian army, had just 36 parts and held 17 rounds in its novel magazine. The result, a lightweight, interchangeable model, captured the imagination of both military buyers and consumers.
The G43 is a single stack, 9mm pistol, and the makers say it’s “ultra-concealable, accurate and fantastic for all shooters regardless of hand size.” In the standard configuration, the gun includes a six-round magazine.
Called the 3DPlus2, this magazine extension was designed in collaboration with Eric Mutchler. Mutchler made headlines earlier this year when he created the Solid Concepts 1911, a 3D printed metal handgun.
The 3DPlus2, printed using a carbon-filled nylon material that Sintercore says is “not only durable but stronger than the pistol’s factory magazines,” adds additional rounds to the Glock 43 factory magazine.
Sintercore says the design is ergonomically correct, easy to install, and that no screws or fasteners are needed to install the device. The company says the magazine extension “simply snaps into place, working with the gun’s factory spring.” They add that it’s also easy to remove due to a hole in the bottom of the extension allows for compression of the factory spring.
According to the company, the extensions will be ready for shipment as of August, and pre-orders are being taken today for $19.95.
The Sintercore extension add two rounds to the Glock 43’s capacity up to 8+1 rounds and extends lower than the standard baseplate. The company says 3D printing technology was critical to the process, not just as it relates to rapid prototyping and proof-of-concept concerns, but because design to production time is radically shorter.
Sintercore says the 3DPlus2 features a correct, ergonomic angle, works with the factory magazine spring and snaps into place with no screws or fasteners.
What do you think of the impact 3D printing has had on gun and gun accessory manufacturing? Let us know in the Sintercore forum thread on 3DPB.com.
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Prior to becoming an member of the U.K. ASICS FrontRunner team, I previously purchased the Accelerate Jacket in Thunder Blue. But once I joined the programme I was given the same jacket in Jungle Green, which was a great bonus as I already was a fan of the jacket.
It's a lightweight hooded jacket that is wind proof and waterproof. Internally you will have slight moisture build-up on the main fabric as you sweat, but this is something that would occur for any fully waterproof jacket.
The jacket fits well, and the adjustable cords on the hood and sides of the lower jacket hem, help give a perfect fit. Particularly when it's raining I've found the adjuster keeps the hood tight to my face, whilst not restricting my movement and providing comfort.
The jacket has a full zip at the front, and the attention to detail with the zipper cover at the top helps prevents irritation on your neck and chin. Additionally there are two good sized zipped hand pockets to place your essentials like smartphone, keys, gels etc.
The two colours have good visibility during the day, furthermore there is a reflective panel on the hood, which make you easily visible in low lighting.
Following purchase of the original jacket, both items remain a staple of my running collection. To summarise this jacket provides good protection against the elements, fits well, and has more than adequate storage for my essentials, price wise it's sits in the middle of comparable jackets and offers really good value for money. I'd recommend it, so much so I bought my wife the same jacket.
Quantity Surveyor from London
Coach: John Starrett

domain: hobby


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Helicopter engine manufacturer Turbomeca and aircraft leasing firm Milestone Aviation Group yesterday at Heli-Expo 2012 inked a partnership on a fixed-hourly-cost maintenance program for Milestone customers flying Turbomeca-powered helicopters.
“Through the Global Support Partnership, Turbomeca and Milestone are jointly developing new engine support solutions and services for Milestone’s lease customers around the world,” the companies said. The first support program to be developed is the Milestone Support by the Hour program, which will provide “added value and other ancillary benefits to Milestone’s customers.” Turbomeca will manage this global support partnership using its worldwide network of support sites and numerous affiliate maintenance centers. “We are excited to announce this strategically important collaboration with Milestone,” said Turbomeca chairman and CEO Olivier Andries. “Milestone presents a new opportunity for us to expand our customer base.”
Milestone chairman Richard Santulli said the partnership “will help a large segment of our customers to control their operating costs.” He added that the announcement “is only the beginning.” Milestone COO Matthew Harris told AIN that his company intends to “provide more solutions” with Turbomeca, as well as additional OEMs and suppliers, “to continue to look for ways to add value for our customers.”

domain: economics


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Eva was sure she looked perfect that evening!
Eva Longoria was also present at the 95th Oscars. The actress appeared in an absolutely gorgeous dress with a daring neckline.
The actress is rarely invited to red carpets and social events, but whenever she appeares at any of them, she always shines on the red carpet. Eva worked on every detail of the dress together with her stylist. This time she was wearing a champagne-colored Zuhair Murad dress that perfectly suited her. The bohemian dress instantly caught the attention of all the photojournalists.
The maxi skirt was good after being lower virtually to the navel. The designer, recognized for his beautiful outfits, managed to maintain her steadiness on a slim line and left nothing else to spoil the boldness of the outfit.
Eva posed for the photographers absolutely conscious of her perfection for the night.

domain: entertainment


text:
Currys bounced higher as it became the latest firm to be caught up in takeover speculation.
Shares in the FTSE 250 electronics retailer jumped 8 per cent, or 7.45p, to 100.3p after financial news blog Betaville suggested a mystery buyer, possibly private equity, was circling the group.
Currys shares have fallen by around a third over the last 12 months, meaning the firm could potentially be eyed as a cut-price acquisition opportunity.
Takeover rumours: Shares in Currys jumped 8.4% after financial blog Betaville suggested a buyer, possibly private equity, was circling the group
It is the second major company to be drawn into takeover gossip since the end of last week, when shares in Rolls-Royce (up 4.1 per cent, or 4p, to 102.38p) soared nearly 20 per cent on similar rumours before dropping back sharply on Monday when additional information on a possible bid failed to emerge.
Richard Hunter, head of markets at Interactive Investor, noted that the UK market had ‘increasingly become an attractive investment destination for overseas buyers’, with bids flowing in for multiple companies including chemist chain Boots.
‘As such, an approach [for Currys] from a value-seeking investor cannot be ruled out,’ Hunter said.
The FTSE 100 was up 0.9 per cent, or 64.11 points, at 7537.25 and the FTSE 250 climbed 2 per cent, or 421.94 points, to 21491.97.
The blue-chip index managed to regain all the ground lost since the start of the war in Ukraine amid hopes of a breakthrough in peace talks as the Russian side showed signs of softening previous demands.
Travel stocks gained on the back of the growing optimism, with British Airways-owner IAG up 2.5 per cent, or 3.4p, to 140.72p, easyJet rose 4.4 per cent, or 22.6p, to 539p and Wizz Air ascended 4.2 per cent, or 106p, to 2658p.
‘The resilience of global stocks given the cocktail of risks facing the global economy is truly impressive,’ said AJ Bell investment director Russ Mould.
However, he warned that the market would face ‘continuing tests’ as the impact of price inflation and interest rate hikes by central banks began to pile on top of the ongoing geopolitical crisis.
Hopes of a diplomatic breakthrough helped lift shares in Ukraine-focused iron miner Ferrexpo, which gained 15.8 per cent, or 26.6p, to 194.5p.
It also updated on its humanitarian efforts in Ukraine, announcing that it had helped provide accommodation and assistance to over 1,400 refugees displaced by the Russian invasion.
Meanwhile, troubled gold miner Polymetal soared 36.9 per cent, or 65p, to 241p after confirming it was ‘evaluating various options’ to change its structure.
The admission followed reports earlier this week that the group was planning to split off its operations in Russia and Kazakhstan to protect itself from sanctions.
Oil prices headed lower with Brent crude dropping below $105 a barrel amid concerns new Covid-19 lockdowns in China would hit demand.
The decline pushed shares in BP down 2.6p per cent, or 10.2p, to 381.5p and Shell fell 1.9 per cent, or 39.5p, to 2022p.
Computer game maker TinyBuild jumped 4.5 per cent, or 6.5p, to 150p after posting full-year results that were better than expected.
The group reported a pre-tax profit of £9.5million, up 63 per cent year-on-year, while revenues jumped 39 per cent to £39.7million.
It also remained ‘confident’ about the coming year despite the disruption caused by the conflict in Ukraine.
AstraZeneca shares hit another record high, rising 1.5 per cent, or 152p, to 10046p after good news for Ondexxya.
Japan approved the drug that stops uncontrolled bleeding in adults taking medicines that prevent blood clotting.
Coca-Cola HBC, the drinks bottling giant which faced backlash over its decision not to exit its Russian business, added 5.8 per cent, or 92p, to 1681.5p as investors hunted for bargains.

domain: news


Optical laser displacement sensors are employed in front of the inlet for panels on straightening machines to prevent damage to the machine. The sensors enable the detection of double sheets and an accurate acquisition of upright edges.
919 787 9707
HEADQUARTER MICRO-EPSILON AMERICA8120 Brownleigh Dr.
Raleigh, NC firstname.lastname@example.org 787 9707919 787 9706


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2018 Idaho Statutes
Title 54 - PROFESSIONS, VOCATIONS, AND BUSINESSES
Chapter 10 - ELECTRICAL CONTRACTORS AND JOURNEYMEN
Section 54-1010 - INSTALLATIONS BY ELECTRICAL CONTRACTOR PERFORMED BY LICENSED JOURNEYMAN — PRIOR CERTIFICATE HOLDERS ENTITLED TO LICENSE — LIST OF ELECTRICIANS IN CONTRACTOR’S EMPLOY.
54-1010. INSTALLATIONS BY ELECTRICAL CONTRACTOR PERFORMED BY LICENSED JOURNEYMAN — PRIOR CERTIFICATE HOLDERS ENTITLED TO LICENSE — LIST OF ELECTRICIANS IN CONTRACTOR’S EMPLOY. (1) Any electrical contractor who works as a journeyman electrician, as herein defined, shall be required to have a journeyman electrician’s license or master electrician’s license issued under the provisions of this act. All installations of electrical wiring, equipment or apparatus made by an electrical contractor shall be done by or under the direct supervision of a licensed journeyman electrician or licensed master electrician.
(2) The individual owner of an electrical contracting business may act as his own journeyman electrician or master electrician provided that he has complied with the provisions of section 54-1002, Idaho Code, pertaining to journeyman electrician. Each electrical contractor in this state shall, upon request of the administrator or his authorized agent, furnish a list of journeyman electricians in said electrical contractor’s employ.
(3) Any individual working as an apprentice electrician, as defined in this act, must be registered with the division of building safety as an apprentice electrician, as provided in section 54-1007, Idaho Code; and it shall be unlawful for an individual to work as an apprentice electrician without possessing a current apprentice registration certificate.
(4) Any individual working as a limited electrical installer trainee, as defined in this chapter, must be registered with the division of building safety as a limited electrical installer trainee. It shall be unlawful for an individual to work as a limited electrical installer trainee without possessing a current registration certificate.
[54-1010, added 1961, ch. 311, sec. 13, p. 583; am. 1974, ch. 39, sec. 86, p. 1023; am. 1986, ch. 309, sec. 2, p. 761; am. 1996, ch. 421, sec. 46, p. 1434; am. 1999, ch. 367, sec. 7, p. 973; am. 2018, ch. 208, sec. 10, p. 465; am. 2018, ch. 209, sec. 10, p. 474.]
Disclaimer: These codes may not be the most recent version. Idaho may have more current or accurate information. We make no warranties or guarantees about the accuracy, completeness, or adequacy of the information contained on this site or the information linked to on the state site. Please check official sources.

domain: law


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Canadian electronic duo Purity Ring have announced extensive 2015 tour dates through North America and Europe.
When do Purity Ring tickets go on sale and what's the deal with pre-sale?
Live Nation, SongKick, and American Express pre-sales begin Wednesday, January 28. Each show may be different, but most tickets go on sale to the general public on Friday, January 30.
Purity Ring All Tour Dates and Tickets
Southwold, England, United Kingdom
Stockholm, Stockholms län, Sweden
København V, Denmark
Amsterdam, NH, Netherlands
Brighton, England, United Kingdom
O2 Ritz Manchester
Manchester, England, United Kingdom
Glasgow, Scotland, United Kingdom
Le Trabendo (Parc de la Villette)
Paris, Île-de-France, France
Saint-Josse-ten-Noode, Bruxelles, Belgium
Bristol, England, United Kingdom
London, England, United Kingdom
For Purity Ring's latest music, news, and tour dates, check out their Zumic artist page.

domain: geography


While the rest of the world is trying to live more sustainably and find ways to reduce its trash output, LG Labs has put its resources into designing yet another pod-based instant coffee machine with a twist: it actually uses two pods to make a single cup of coffee, doubling the amount of trash generated with every cup brewed.
We’ve explained time and time again why pod-based coffee machines from companies like Keurig and Nespresso are a bad idea. Sure, they’re more convenient than having to manually lift a spoonful of freeze-dried instant coffee and dump it into a mug, but unless you’re using a refillable pod (based on how many varieties of pre-filled pods are available in stores, most users clearly aren’t filling their own) these machines generate a lot of unnecessary trash. And while both brands have worked to ensure their respective pods are fully recyclable, that doesn’t guarantee that all consumers will be responsible enough to dispose of them properly.
LG Labs has decided that what’s more important for the Earth is giving consumers the chance to feel like they’re coffee shop baristas. The Duobo, which looks like a three-legged alien robot, is designed to use two Nespresso-style pods for every cup of coffee it produces, allowing users to mix flavors to create their own custom blends. Using buttons on the Duobo’s base, or a mobile app that wirelessly connects to the coffee maker, users can also adjust the “extraction temperature” of the water and the amount of pressure the machine uses to fine-tune the brew to their preferences.
Somehow, an alien-like design or churning through twice the number of coffee pods per cup isn’t the Duobo’s weirdest feature. The machine’s base includes an LCD screen that’s used to display custom content including coffee-related PSAs, promotions for coffee-related events, and ads for coffee brands, while the user waits for their beverage to brew. Putting screens on appliances isn’t a new idea, there are lots of fridges with the feature, but it’s confusing how it’s been implemented here, as the Duobo’s display is located right where you’re supposed to place your coffee cup to be filled. The display is going to be obscured every time the machine is used, and it’s doubtful anyone’s going to want to hang around it when they’re not seeking a caffeine fix.
That probably explains why this LG Labs creation is making its way to consumers through a Kickstarter crowdfunding campaign, instead of the company churning out thousands that could collect dust on store shelves. Early bird backers of the campaign can pre-order the Duobo, with delivery expected sometime in December, for a hefty $399, which is actually half the expected full retail price of $799. Oof. With most crowdfunded products there’s the risk of nothing ever materializing, or considerable shipping delays, but here it seems like LG Labs is using the platform as a way to gauge interest in the Duobo so the company has a better idea of how many to manufacture. With just 70 backers so far, it doesn’t seem like Keurig has much to worry about.


Khaleegy (Gulf Region)
This typical women’s dance comes from the Gulf Region, (e.g., Persian Gulf/Arabian Peninsula, Saudi Arabia, Kuwait, Bahrain, Oman, Qatar (United Arab Emirates). Because this dance comes from the Arabic gulf region, the khaleej, explains why it is often called khaleeji but it is not the only dance they have for this region. The locals in the Emirates call it raks na’ashat to distinguish the women’s dance from the various men’s dances. In Kuwait it can be referred to as Samra/Samri and in Saudi Arabia it can be referred to as Saudi. It is said that this dance was “imported”. There are also a variety of dance forms in the Khaleej mostly performed by men (some with a variety of weapons).
Khaleegy is also known as the Hair Dance. The women often dance in groups, toss their hair and gracefully make small gliding steps, but it is much more than a “Hair Dance.” There are a few basic steps that can be explained as “limping” or “gliding”. These steps are used with variations of manipulating their large dresses, executing small shoulder shimmies, head slides and occasional accents, as well as the use of hand gestures. Some hand gestures are very typical while some may have a different meaning. There is very little hip work involved, some hip circles with one hip and small hip drops. Learning raks na’ashat would be beneficial to Oriental dancers as many pieces of Classical music for Raks Sharki include a small Khaleegy part.
The traditional dress for this dance is known as a Thobe Al Nasha’ar. Nasha’ar translated means “to spread out, to unfold, open, or to hoist, thus explaining the various movements executed with the dress and the manner in which it is held. Currently there are two types of Thobe Al Nasha’ar also known as darrah zerri . The traditional style of dress is longer in the back than in the front. The other is a more modern style which is cut floor length all the way around. The traditional style of dress is the one most used for this dance. The under dress for the traditional dress is whatever the women have on at the time to include jeans. The modern version is worn over a matching galabia with long sleeves. For a more theatrical look, you can have a long sleeved galabia from gold or silver fabric so it shines through the thobe as they are somewhat sheer.
The Khaleej is very rich in rhythms and aside from what is referred to as the Khaleegy/Saudi 2/4 rhythm that is commonly used, (Dum Dum teka Dum Dum), there are many others, e.g., Ashory, Emerati, Daza, Bastah, Shara. The most popular songs used “Leyla Leyla” aka “Aba’ad” (Boad) as it is called originally and “Wah Wah” (“And And”).
NOTE: There is a hair dance/trance dance called NACH in North Africa (Tunisia) that is very similar to the khaleegy. The women dance mostly in a kneeling position on the floor.


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VOLKSWAGEN GROUP RAISES INVESTMENTS IN FUTURE TECHNOLOGIES TO EUR 73 BILLION
>> Planning Round 69 includes capex and R&D investments for electrification, hybrid powertrains and digital technology for the next five years
>> Share of investments in future technologies raised to about 50 percent out of the total EUR 150 billion from 40 percent in previous planning round
>> Investments in digitalization to be doubled to around EUR 27 billion
The Volkswagen Group is pressing ahead with its transformation into a digital mobility company. As decided in Planning Round 69, the Group will spend around EUR 73 billion on electrification, hybrid powertrains and digital technology over the next five years. This was announced following today’s Supervisory Board meeting. Investments in Capex and R&D for future technologies will be raised to 50 percent from 40 percent of the Group’s total investments of around EUR 150 billion. Investments in digitalization will double to EUR 27 billion by mid-decade, reflecting the Group’s strong focus on building up software capabilities. Approximately EUR 35 billion will be spent on battery-electric vehicles. A further approximately EUR 11 billion has been earmarked for the development of hybrid vehicles of existing models.
"As part of Volkswagen Group’s investment planning, the Board of Management and Supervisory Board today set the cornerstones for securing the Group’s future success. The transformation of the Group and its brands and the strategic focus on the core areas of mobility will be consistently implemented. Considering the enormous challenges we face in the coming years, our financial basis is very solid," said Hans Dieter Pötsch, Chairman of the Supervisory Board of the Volkswagen Group.
"Having set the course for a battery-electric future in the Volkswagen Group early on, we are now a global leader with our electric platforms and a broad range of electric vehicles," said Herbert Diess, Chief Executive Officer of the Volkswagen Group. "In the coming years, it will be crucial to also reach a leading position in car software in order to meet people’s needs for individual, sustainable and fully connected mobility in the future. To that end, we have doubled our digitalization spend."
Bernd Osterloh, Chairman of the General Works Council, commented: "The investments demonstrate that our Group walks the talk when it comes to transformation, electric mobility and digitalization. We will go fully on the offensive in the coming years. Yet, just as important as billion-euro budgets are a common understanding of the transformation and adequate planning reliability. This is where all of us – employees, managers, the Board of Management and also policy-makers – come in. As regards the latter, I am thinking in particular of Berlin and Brussels and hot topics such as charging infrastructure and high-speed Internet. Volkswagen is fully committed to climate-friendly, highly networked mobility. But for this, we and our customers need a reliable environment."
The Planning Round is based on the expectation that the global economy will grow moderately over the next five years. Moderate growth with regional differences is also anticipated for the individual markets. The increase in productivity of 30 percent and savings in administration are to secure the transformation financially. In addition, the Group is systematically working on the optimization of its product portfolio. Thus, model variants, engine-transmission combinations and features that are less in demand will be streamlined in order to reduce the complexity and increase the efficiency of the portfolio. The planning excludes the joint venture companies in China, since they are not part of the consolidated group and finance investments in plants and products from their own resources.
Electric mobility and hybrid technologies
Volkswagen was the first automaker to commit to the Paris Agreement and aims to become climate neutral by 2050. Over the next ten years, the Group intends to launch approximately 70 all-electric models by 2030. Around 20 of these are already in production, with 50 more to follow. In addition, around 60 hybrids are planned by the end of the decade, slightly over half of which are already being manufactured.
The Planning Round envisages production of approximately 26 million fully electric cars by 2030. Some 19 million of these vehicles will be based on the Modular Electric Drive Toolkit (MEB), with most of the remaining seven million to use the high-performance PPE platform. The Group estimates production of around seven million hybrid vehicles over the same period.
Significantly greater emphasis on the key topic of digitalization
Due to the crucial role that software will play for Volkswagen’s future success, capital spending on digitalization will total around EUR 27 billion, double the amount set in the previous Planning Round. This includes spending on the Group’s Car.Software organization, which was successfully launched earlier this year. The goal is to build a proprietary software stack, which will be deployed in Audi’s Artemis project from 2024. The company's own share in software is to increase from 10 to 60 percent. In addition, a large share of the funds earmarked for digitalization will be invested in the mission-critical fields of artificial intelligence, autonomous driving and digitalization of significant business processes.
Transformation of Germany as a center of industry to be pushed
By investing in its German sites, Volkswagen, as one of the country’s largest industrial players, is contributing to a strong German economy,while continuing to push the country’s transformation into a more sustainable and more digital future with long-term job security. After Saxony, Lower Saxony will become Germany’s second center of electric mobility.
In Salzgitter, the Group will invest around one billion euros in the strategically important battery technology. In conjunction with its Swedish joint venture partner Northvolt, Volkswagen will build a cell production facility there that will start operation in 2024 and increase the competitiveness of Germany as an industrial location. This is a further key element in the successful transformation of the Volkswagen Group Components division, initiated back in 2015.
At the individual locations, the strategy of consolidating product families at multi-brand plants will be continued in order to leverage even more synergies between the brands and achieve further efficiency gains.
Wolfsburg will add a model to its portfolio: another SUV targeting the European market will roll off the assembly line from 2024. The site already produces the entire Golf family with all its derivatives, the Volkswagen Tiguan and the SEAT Tarraco.
In future, Volkswagen Commercial Vehicles will build the all-electric ID. BUZZ at its Hanover location, along with three fully electric D-SUV models for other Group brands. The Hanover site is thus accelerating the transformation to electric mobility that has already commenced.
At the Volkswagen’s brand plant in Emden, the transformation is also progressing rapidly: while the construction work for the switch to electric mobility is fully on track, the second electric model for the site has now been defined. In addition to the ID.4, the four-door all-electric Volkswagen Aero is scheduled to be manufactured there from 2023.
At the same time, the Passat currently built in Emden will be produced in Bratislava, Slovakia, starting in 2023 – together with the ŠKODA SUPERB family – which will leverage synergies within the vehicle family here, too. Moving production of the SUPERB from the Czech plant in Kvasiny to Bratislava will give ŠKODA the necessary capacity in the Czech Republic to carry out the brand’s growth plan.


Building on the legacy of the original, first-generation SCCA Trans Am-series contender, the 2015 Camaro Z/28 is our most track-capable model in its history. And now you can add many of its high-performance parts to your gen-five Camaro. Majority of the components from our special Z/28 collection are the same factory-engineered and -validated parts used on the production vehicles.
The components are designed for compatibility with Camaro SS. Some additional modifications and fabrication may be required with some components adaptable to V6 models. Check out the entire collection and take your Camaro’s performance to the next level.
The Z/28 is the first high-volume production road car to employ racing-derived DSSV® (Dynamic Suspensions Spool Valve) damper technology from Multimatic.™ The dampers rely upon a pair of self-piloted spool valves to control fluid through tuned port shapes rather than conventional deflected disc dampers. The design of the inverted-monotube front strut and aluminum-body monotube rear hydraulic dampers offers maximum response, stiffness, and tuning optimized for the track, with the highest level of damper predictability, accuracy, and repeatability. The unique dampers are the center of a comprehensive suspension
package that also includes higher-rate coil springs, smaller-diameter solid stabilizer bars, and higher-rate suspension bushings. The dampers are matched with optimized stiff spring rates and stabilizer bars in both the front and rear to reduce body movement in hard cornering and acceleration. Also included in the package are front ride-and-handling links, rear upper control arms, rear trailing links, and toe links which have upgraded stiffer bushings designed to reduce cornering compliance and increase durability on the track.
This kit includes the rear lower control arms and hardware necessary to properly install the stability bars included in the 1LE and Z/28 suspensions. This kit is needed for 2010 and 2011 model year Camaros only, as the size of the stabilizer bar clamp stud changed starting in 2012 for both the front and the rear bar clamps.
The Z/28 high-flow exhaust manifolds will provide increased airflow to the Camaro LS3/L99 6.2L engine. This kit enables bolt-on installation of the Z/28 dual-mode exhaust system. However, it will not work with the production LS3/L99 exhaust system.
Cat-back exhaust kits are a great way to increase power and performance with a simple bolt-on installation. This kit comes complete with pipes, mufflers, hangers, and exhaust tips assembled as a single unit. It features 2.75" pipes, with 26% less back pressure than a stock LS3 system.
Bolts an LS7 Z/28 cat-back exhaust to LS3 Gen-V Camaro. This is the only kit necessary to clamp the LS7 Z/28 exhaust (P/N 22906768) right up to your production LS3-equipped SS. The Z/28 exhaust system provides a 26% reduction in back pressure compared with an LS3-equipped Camaro.
The Camaro Z/28 is equipped with an RDM cooling system that circulates cooled transmission fluid through a heat exchanger integrated into the RDM cover. This innovative system reduces oil temperature by more than 100°F, helping the differential maintain cool, stable performance throughout the most aggressive track driving sessions. The Z/28 system has greater heat-rejection capability than competitors’ air-cooled RDMs, helping to prevent differential component damage from high-temperature oil degradation. The integral heat exchanger design also eliminates the weight, noise, and complexity of a dedicated external pump and its associated hardware, seen in many aftermarket systems. This Z/28 based RDM cooling kit can be applied to any 2010 and later Camaro SS with a manual transmission.
A unique open-air box intake system is used with the Z/28’s LS7 engine, featuring a replaceable conical air filter and delivering the highest airflow performance of any production Camaro filter system. The air cleaner seals around the bottom of the hood, reducing the chance that recirculated hot air will be drawn into the engine. It is a direct replacement for the intake system on the Camaro SS. The Z/28 air intake provides a 51% reduction in air restriction when compared with a standard LS3-equipped Camaro. Includes hardware that supports 1LE and SS PCV systems. Please note: This product is non-emission compliant and cannot be sold in the state of California. For off road use only. Not recommended for use on L99-equipped Camaros.
Brake ducts decrease “brake fade,” increase brake component life, create secondary cooling of brake fluid, and sustain brake pedal feel for longer periods of time. This is completed by a supply of constant fresh air to the back of rotor area. This air supply improves convection of the brake rotor vanes, reducing heat build up compared with conventional airflow, thus increasing brake performance. Fits 2014-2015 SS- and 1LE-equipped Camaros.
The rear spoiler kit contains everything you need to boost the downward force you’re looking for. The wicker blade spoiler extension rigidly attaches with 7 bolts to help increase durability. The spoiler and extension wicker can add more than 110 lbs at speeds above 125 mph. The kit includes instruction sheet, rear wicker extension, installation drill template, and attaching fasteners. Compatible with the Rear Blade Spoiler (sold separately).
Select components from the Z/28 Camaro’s downforce-enhancing exterior are offered in this package of bolt-on components, including a front splitter and wheel flare moldings that fit a rear spoiler with a wicker bill. The wheel flare moldings are designed to cover wide tires and feature deflectors at the bottom-front corners that contribute to downforce-producing aerodynamics, while the unique rear spoiler’s wicker bill is a small, vertical tab at the top edge. On the Z/28, it adds approximately 28 counts of drag, improving rear lift performance by 70 counts. That allows the Z/28 to handle turns at higher speeds and delivers greater overall high-speed stability.
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] [Thread Prev
Re: o.t. READING
The most recent things I've read (besides the stack of Electronic Musician, Mix, etc...)
American Gods - Neil Gaiman
The Berserker Wars - Fred Saberhagen
Time Enough for Love: The Lives of Lazarus Long. - Robert Heinlein
The Nightmare Factory - Thomas Ligotti. (Great creepy stuff if you like Lovecraft)
Then there's the 1/2 dozen books on fly fishing that get read a chapter here and there.

domain: library


|Li Keqiang and Prime Minister of Serbia Jointly Attend Completion Ceremony of Bridge Across Danube River in Belgrade, Stressing to Build Friendship Bridge Linking Hearts of Chinese and Serbian Peoples and Cooperation Bridge Bringing Mutual Benefits and Win-win Results for China and Europe
Premier Li Keqiang and Prime Minister Vučić then jointly unveiled the monument marking the completion of the bridge……
|Li Keqiang Arrives in Belgrade to Attend Meeting of Heads of Government of China and Central and Eastern European Countries and Pay an Official Visit to Serbia
In the evening of December 15 local time, upon the invitation of Prime Minister Aleksandar Vucic the Republic of Serbia……
|Li Keqiang Attends Third Meeting of Heads of Government of China and Central and Eastern European Countries
In the afternoon of December 16 local time……
|Li Keqiang Holds Talks with Prime Minister of Serbia, Stressing Deepening of Mutually Beneficial Cooperation to Realize Common Development and Promotion of China-Serbia Strategic Partnership to a New Level
In the afternoon of December 17 local time, Premier Li Keqiang held talks with Prime Minister Aleksandar Vučić of Serbia in Belgrade……
|• Chinese premier hails bright prospect of infrastructure cooperation with CEE countries
Chinese Premier Li Keqiang said Thursday China and Central and Eastern European(CEE)countries enjoy broad prospect in their cooperation on infrastructure construction.
|• Premier Li receives Belgrade honorary citizen charter
Chinese Premier Li Keqiang(R)receives the Belgrade honorary citizen charter at the Belgrade City Assembly in Belgrade, Serbia, Dec.18.
|• China, CEE countries eye land-sea express passage to speed up delivery
Chinese Premier Li Keqiang(2nd R)meets with Serbian Prime Minister Aleksandar Vucic(2nd L), Hungarian Prime Minister Viktor Orban(R)and Macedonian Prime Minister Nikola Gruevski in Belgrade, Serbia...
|• The Belgrade Guidelines for Cooperation between China and Central and Eastern European Countries
On 16 December 2014, the 3rd Meeting of Heads of Government of China and Central and Eastern European Countries was held in Belgrade, Serbia.
|• China pledges 3-billion-USD investment fund for CEE countries
Chinese Premier Li Keqiang said Tuesday that his country will create an investment fund of 3 billion U.
|• China, CEE countries pledge closer cooperation on interconnectivity
China and the Central and Eastern European(CEE)countries on Tuesday agreed to enhance cooperation on interconnectivity, according to fresh guidelines issued following the third leaders'meeting betw...


76ers trade Furkan Korkmaz to Pistons
The Pistons appear willing to trade Saddiq Bey ahead of the trade deadline. According to James L. Edwards III of the Athletic, he might even be a more likely trade candidate the Bojan Bogdanovic or Alec Burks.
For the Sixers, this trade accomplishes two things: it nets them a rotation-caliber wing in Bey, and it sheds salary to help save ownership a few tax dollars. Philadelphia has a reported interest in shedding salary and a reported interest in getting better. In this trade, they accomplish both.
Bey has struggled with his efficiency over the years, but he’s a capable volume shooter and multi-position defender on the wing. One has to imagine his shooting numbers might creep in the right direction with a more streamlined role playing off of Joel Embiid and James Harden.
At 23 years old, Bey is in the third year of his rookie contract and is due $4.5 million next season. That’s a small price for a rotation player with upside. Bey would give the second unit more juice offensively and he’s more postseason-friendly, at least on paper, than Matisse Thybulle or Georges Niang.
The Pistons, meanwhile, get second-round picks (including the Charlotte pick, which figures to land very low in the 30s), a flier on Korkmaz, and Jaden Springer, the Sixers’ most recent first-round pick.


text:
Courtney Flatt, Northwest Public Radio
The federal government is standing by its previous plans for managing the Columbia River to prevent the extinction of its salmon and steelhead. That means little would change for dam operations on the West’s biggest river — but only if it wins court approval.
Officials Friday released the finalized plan, known as the biological opinion or BiOp. It guides dam operations to assure they do not lead to the extinction of 13 species of salmon and steelhead that are protected under the Endangered Species Act. The plan has been the subject of more than 20 years of legal conflict between people who want to protect salmon and people who want the dams to produce hydroelectricity and maintain shipping traditions.
The National Oceanic and Atmospheric Administration is the lead agency in developing the biological opinion. It says the current plan is on track to meet Endangered Species Act goals for the federally protected fish. NOAA officials say the plan may better protect some fish than previously thought.
“The actions outlined in the biological opinion, and the operation of the hydro system, is designed to move us in the direction towards recovery and avoid jeopardy, and this program does that,” said NOAA’s Barry Thom. “It actually does improve the status of the populations over time. But it is not designed to achieve ultimate recovery of the population.”
Officials say the 610-page plan will protect and improve habitat, with specific attention paid to tributaries and estuaries of the Columbia and Snake rivers.
“A major focus of the tributary habitat program is to help us buffer against potential effects of climate change in the system, so that the habitat projects … are designed to maintain and protect the cool water inputs into the system,” Thom said during a conference call with reporters.
NOAA released a draft version of this plan in September.
In 2011, U.S. District Judge James A. Redden rejected the plan and asked the Obama administration to consider more ways to recover the endangered fish.
Redden’s suggestions included spilling more water over the dams to help juvenile salmon safely make it downriver to the ocean, changing reservoirs to help fish passage, and removing the Snake River dams altogether.
The case has been transferred to Judge Michael H. Simon. He has yet to set a court date for the plan’s sponsors and opponents to argue it. He’ll then decide if the plan is adequate to protect Columbia River salmon and steelhead.
Now that the previous version of the plan is partway completed, supporters say a trend toward larger salmon runs shows the plan is working. Terry Flores is with Northwest RiverPartners, which represents commerce and industry groups that defend the presence of hydroelectric dams on the Columbia-Snake system.
“This plan is amazing. It’s the most comprehensive plan we can find anywhere in this country by far,” Flores said.
Environmental groups say they are disappointed with this finalized plan. Gilly Lyons, with advocacy group Save Our Wild Salmon, said the group is frustrated.
“The federal agencies in charge here have re-isuued a slightly tweaked, but largely status quo federal salmon plan that repeats a lot of the same mistakes over the past decade or so that kept them in court and bound them in litigation over these dams and the salmon that they impact,” Lyons said.
Lyons said it is too soon to tell if environmental groups will file another lawsuit.
“With all the stuff that we see in the plan, or that’s not there, as the case may be, it sure looks like the federal government would like to go back to court,” Lyons said.
NOAA officials said there will be a few years before they have to start writing a new 10-year plan beyond 2018.
“One main priority is to carry out this existing biological opinion,” Thom said. “There will be a period of time between [the 2018 discussions] and the next couple of years where I would like to focus our efforts on talking about long-term recovery. … As opposed to focusing either on A) the litigation or B) the details of a new biological opinion beyond 2018.”


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Open Access This article is
- freely available
Molecules 2018, 23(2), 333; doi:10.3390/molecules23020333
The Fungal Metabolite Eurochevalierine, a Sequiterpene Alkaloid, Displays Anti-Cancer Properties through Selective Sirtuin 1/2 Inhibition
Laboratoire de Biologie Moléculaire et Cellulaire du Cancer, Hôpital Kirchberg, 9, rue Edward Steichen, L-2540 Luxembourg, Luxembourg
Department of Pharmacotherapy and Pharmaceutics, Faculté de Pharmacie, Université Libre de Bruxelles, Boulevard du Triomphe, 1050 Brussels, Belgium
Service de Neurochirurgie, Hôpital Erasme, Université Libre de Bruxelles, 808 Route de Lennik, 1070 Brussels, Belgium
Department of Pharmacy, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, 1 Gwanak-ro Gwanak-gu, Seoul 08826, Korea
Dipartimento di Scienze Chimiche, Università di Napoli Federico II, Via Cintia 4, 80126 Napoli, Italy
Instituto de Ciências Biomédicas de Abel Salazar (ICBAS) and CIIMAR, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal
College of Pharmacy, Chung-Ang University, 84 Heukseok-ro Dongjak-gu, Seoul 06974, Korea
Retired – previously Director of Research at the Fonds National de la Recherche Scientifique (FRS-FNRS; Belgium)
Authors to whom correspondence should be addressed.
Received: 12 January 2018 / Accepted: 31 January 2018 / Published: 5 February 2018
NAD+-dependent histone deacetylases (sirtuins) are implicated in cellular processes such as proliferation, DNA repair, and apoptosis by regulating gene expression and the functions of numerous proteins. Due to their key role in cells, the discovery of small molecule sirtuin modulators has been of significant interest for diverse therapeutic applications. In particular, it has been shown that inhibition of sirtuin 1 and 2 activities is beneficial for cancer treatment. Here, we demonstrate that the fungal metabolite eurochevalierine from the fungus Neosartorya pseudofischeri inhibits sirtuin 1 and 2 activities (IC50 about 10 µM) without affecting sirtuin 3 activity. The binding modes of the eurochevalierine for sirtuin 1 and 2 have been identified through computational docking analyses. Accordingly, this sequiterpene alkaloid induces histone H4 and α-tubulin acetylation in various cancer cell models in which it induces strong cytostatic effects without affecting significantly the viability of healthy PBMCs. Importantly, eurochevalierine targets preferentially cancer cell proliferation (selectivity factor ≫ 7), as normal human primary CD34+ stem/progenitor cells were less affected by the treatment. Finally, eurochevalierine displays suitable drug-likeness parameters and therefore represent a promising scaffold for lead molecule optimization to study the mechanism and biological roles of sirtuins and potentially a basis for development into therapeutics.
Keywords:cancer; epigenetics; HDAC; sirtuin inhibitor; natural compound; eurochevalierine; cytostatic compound
The European Union  and the United States of America (USA)  as well as most developed countries are facing an increasing health problem with respect to cancer. In the European Union, cancer represents the second most important cause of death and morbidity with one out of four deaths . In the USA, about 1.7 millions new cancer cases and about 610,000 cancer deaths are projected for 2018 .
There is increasing evidence that, besides genetic lesions, epigenetic alterations including DNA methylation, small non-coding RNAs, and histone modifications are regulating essential steps of carcinogenesis [3,4,5,6]. Among the enzymes responsible for regulating epigenetic marks, histone deacetylases (HDACs), the enzyme responsible for the removal of lysine acetyl groups, became an interesting target for anti-cancer therapy [7,8]. There are 18 members in the HDAC family classified into four major groups. Class I, II, and IV HDACs are zinc-dependent, whereas Class III HDACs, also called sirtuins (SIRTs), require nicotinamide adenine dinucleotide (NAD+) as a co-factor [9,10]. Sirtuins (SIRT1-7) share extensive homologies with the yeast HDAC Silent Information Regulator 2 (Sir2). SIRT1 is essentially nuclear and modulates acetylation of histone H1, H3, and H4, whereas SIRT2 is predominantly cytoplasmic and regulates α-tubulin, together with HDAC6 . HDAC inhibitors (HDACi) of class I, II, and IV are the most widely used for basic research as well as in the clinic [12,13]; however, sirtuin inhibitors (SIRTi) represent a novel, attractive approach for anti-cancer drug development [6,7]. Hence, numerous reports demonstrated that SIRT1, SIRT2, or dual SIRT1/2 inhibitors may generate anticancer effects. However, depending on the inhibitors and cancer cell models, the observed antitumor effects and cellular outcomes are not always comparable and can be mediated by various mechanisms . For instance, the SIRT1/2 inhibitor sirtinol triggers growth arrest in human breast and lung cancer cells . Similarly, the benzopyran-based compound 18 inhibits SIRT1/2 activities and acts as a cytostatic agent in glioblastoma cells , whereas the dual SIRT1/2 inhibitor tenovin-1 induces apoptosis in sarcoma cells . Other specific sirtuin inhibitors (SIRTi) have been reported to trigger cancer cell death, including the SIRT1 inhibitors NCO-01 and -04, and the SIRT2 inhibitors AEM1 and AEM2, which induce cell death in T-cell leukemia/lymphoma  and in non-small cell lung cancer , respectively. Nevertheless, several reports have demonstrated that the knockdown of SIRT1, SIRT2, or SIRT1/2 genes affected less cancer cell viability but essentially proliferation and/or invasion .
Natural products and their semisynthetic or synthetic derivatives represent more than 50% of the current armamentarium to combat cancer [20,21,22,23,24,25]. Various natural compounds display anti-HDAC activity [8,10,26] and the current study details the anti-HDAC activity of eurochevalierine, a sequiterpene alkaloid (Figure 1), which is a fungal metabolite isolated from Eurotium chevalieri Mangin that is an imperfect stage of Aspergillus chevalieri (Mangin) Thorn and Church and that belongs to Phylum Ascomyceta . We previously showed that eurochevalierine is a cytostatic but not cytotoxic compound in vitro with respect to the growth of various cancer cell models . Eurochevalierine does not induce apoptosis in cancer cells and displays its cytostatic effects even in apoptosis-resistant cancer cells .
The aim of this work was to further investigate epigenetic regulatory mechanisms that could account for the observed inhibition of cancer cell proliferation. Based on computational docking experiments combined with in vitro HDAC activity assays and the examination of the profile of acetylation of α-tubulin and several histone marks in various solid cancer models, we established that eurochevalierine leads to specific inhibition of SIRT1 and 2.
We previously published a series of secondary metabolites from Neosartorya pseudofischeri, among which eurochevalierine (Figure 1) displayed interesting anti-cancer properties against multiple cell lines of various origins . More recently, another group further confirmed the relevant cytostatic activity of eurochevalierine in glioblastoma and non-small cell lung cancer cells . Accordingly, in the current study, we sought to further investigate the anti-cancer properties of this sequiterpene alkaloid. Although eurochevalierine displays cytostatic effects in various cancer models, we did not observe eurochevalierine-induced modifications in the cell cycle distribution of cancer cells as revealed by flow cytometry analyses. Nevertheless, a shift of the DNA histograms was observed in eurochevalierine-treated A-549 cancer cells as if treated cells integrated more propidium iodide than control cells (Figure 2A). DNA histogram analyses also revealed that eurochevalierine did not increase the sub-G1 fraction in A-549 lung cancer, U-373 glioblastoma, or B16-F10 melanoma cells, even after 72 h of treatment with a concentration as high as 50 µM (Figure 2B), which nearly corresponds to two times the mean IC50 value determined in a panel of cancer cell lines regarding growth inhibition . These results confirmed that eurochevalierine does not behave as a cytotoxic compound and exerts its anti-cancer activity via cytostatic properties.
Previous studies already revealed that the DNA conformation  and modifications in histone H1 concentration  influences DNA stainability by propidium iodide as revealed by flow cytometry analyses. It is well established that histone modifications including lysine acetylation and its regulation by HDACs exert major roles in DNA structure [3,26,32]. Thus, we evaluated the effect of eurochevalierine on in vitro HDAC activity. Eurochevalierine inhibited only about 33% of total HDAC activity at a concentration of 100 µM (Figure 3A).
In order to determine whether this effect results from a weak inhibition of multiple HDAC isoenzymes or a strong inhibition of a limited number of selected isoforms, we further tested the effect of eurochevalierine on the deacetylase activity of selected HDAC members. At the concentration of 100 µM, eurochevalierine remained without effect on HDAC1, 2, 8, 10 and 11, inhibited about 36 and 51% of HDAC6 and 11 activities, respectively, and induced HDAC3 activity by 1.5-fold (Figure 3B). Interestingly, eurochevalierine had IC50 values of 9.8 and 10 µM against SIRT1 and 2, respectively, whereas it failed to reach the IC50 for SIRT3 at 100 µM (Table 1). Such values are comparable to the one obtained with suramin used as a potent positive control for in vitro SIRT1 and SIRT2 inhibition with IC50 values of 2.8 and 13 µM, respectively, and was inactive against SIRT3, which is in agreement with previously published studies . The natural SIRT inhibitor nicotinamide was also included as a reference compound for in vitro SIRT3 inhibition . Besides suramin, additional references SIRT1/2, SIRT1, and SIRT2 inhibitors, namely, sirtinol, EX-527, and AGK2, respectively, performed in a similar range or less efficiently .
To further explore the potential of eurochevalierine as an SIRT inhibitor, we compared the docking affinity between eurochevalierine and known reference inhibitors (sirtinol, EX-527, and AGK2) using AutoDock Vina . A carboxamide derivative and SirReal2 were used as control docking molecules for SIRT1 and 2, respectively, to validate our docking experiments. The control docking results were very close to the actual binding mode and the overall root-mean-square deviation (RMSD) values between the actual binding mode and simulated docking pose of the carboxamide derivative on SIRT1 and of SirReal2 on SIRT2 were 1.3 Å and 0.4 Å, respectively (Supplementary Materials Figure S1). Qualitative molecular docking experiments were performed with eurochevalierine in comparison with sirtinol and EX-527 against three available human SIRT1 crystal structures that contain two static water molecules in the binding site that would be influential both in the actual inhibition and in the docking experiment [36,37]. The resulting lowest docking affinities are shown in Table 2.
In Figure 4a, the docking pose of eurochevalierine with the lowest binding affinity was selected as the most favorable binding mode in the binding pocket of SIRT1 [PDB code 4ZZI], and the carboxamide derivative, the inhibitor in the original SIRT1 crystal structure, was superposed for comparison . Even though the binding affinity for eurochevalierine was higher than two reference molecules, sirtinol and EX-527, eurochevalierine was predicted to interact with SIRT1 primarily through hydrophobic effects and to occupy the hydrophobic cage formed by residues Ala262, Phe273, Phe297, Ile347, Ile411, Val412, Phe413, and Phe414, as the control molecule binds to SIRT1. Notably, eurochevalierine exhibits π–π stacks with the side chain of Phe273 like the carboxamide derivative.
In addition, similar docking experiments were performed with eurochevalierine in comparison with sirtinol and AGK2 against three available human SIRT2 crystal structures that contain one static water molecule in the binding site [38,39]. The resulting lowest docking affinities are shown in Table 3. The calculated binding affinity could not correlate our experimental data, such as the IC50 values in all aspect, which is mainly caused by the limitation of qualitative docking simulations such as protein flexibility, water/buffer treatments, and the less specific prediction of hydrophobic effects. In Figure 4b, the docking pose of eurochevalierine with the lowest binding affinity was selected as the most favorable binding mode in the binding pocket of SIRT2 (PDB code 4RMH), and SirReal2, the inhibitor in the original SIRT2 crystal structure, was superposed for comparison . Eurochevalierine was predicted to occupy the pocket adjacent to the NAD+-binding site like SirReal2 (ref. accession number: 25672491) and to interact with SIRT2 primarily through hydrophobic effects stabilized by the hydrophobic cage (residues Phe96, Ile118, Phe119, Phe131, Ala135, Leu138, Tyr139, Pro140, Phe143, Ile169, Phe190, Ile232, Val233, and Phe234).
To assess the HDAC inhibitory activity of eurochevalierine in cellulo, we analyzed the acetylation status of SIRT1 and 2 substrates including histone H4, histone H3 at lysine 56 and α-tubulin in SK-MEL-28, U-373, and A-549 cell lines. Since suramin, an SIRT1 and 2 inhibitor reported in the literature, failed to induce any increase in histone or α-tubulin acetylation in our models (Figure S2), cells treated with the pan-HDACi suberoylanilide hydroxamic acid (SAHA) served as a positive control for the induction of protein acetylation. Eurochevalierine induced a robust and dose-dependent increase in the acetylation of sirtuin targets (Figure 5), in agreement with the in vitro inhibition of SIRT1 and 2 activities.
As a potential candidate for further anti-cancer drug development, we then evaluated the biological activity of eurochevalierine in healthy cell models. First, we tested the effect of eurochevalierine on the viability of peripheral blood mononuclear cells (PBMCs) from healthy donors treated for up to 48 h. Results (Figure 6) revealed that eurochevalierine does not trigger any acute toxicity on this model even at a concentration of 100 µM.
Since eurochevalierine exerts its anti-cancer activity as a cytostatic agent, we further tested the impact of this natural compound on a proliferating healthy cell model represented by primary human CD34+ stem/progenitor cells. We observed a maximum growth inhibition of around 30% (Figure 7A) upon 200 µM eurochevalierine even after 72 h of treatment associated with a minor impact on cell viability (Figure 7B). Considering that the mean IC50 value for the growth inhibition of cancer cells is 28 µM , and that, in health cells, a concentration of 200 µM is not sufficient to reach the IC50 (Figure 7A), eurochevalierine displays an important selective toxicity (factor >> 7) for cancer cells compared to normal proliferating cells.
Finally, we wanted to evaluate the drug-likeness properties of eurochevalierine in comparison to other reference SIRTi. Accordingly, we checked whether compounds obey to the extended “rule of five” [molecular weight comprised between 180 and 500 kDa, miLogP ≤ 5, rotatable bonds ≤ 10, topological polar surface area (TPSA) ≤ 140, hydrogen-bond donors ≤ 5 and hydrogen-bond acceptors ≤ 10) combined to an in silico assessment of ADMET (absorption, distribution, metabolism, elimination, and toxicity)] properties (Table 4 and Table S1).
Collectively, such predictive analyses indicate that eurochevalierine is a suitable drug candidate with more favorable parameters compared with other reference SIRT inhibitors, exhibiting high probability of good oral bioavailability and absorption and a low risk of toxicity. Indeed, the number of rotatable bonds in the eurochevalierine molecule, which defines the molecular flexibility for membrane permeation, was just above the defined limit, and the favorable topological polar surface area, which predicts drug transport properties, is in agreement with a favorable intestinal absorption parameter (92.6%). Unlike eurochevalierine, other compounds such as sirtinol, EX-527, and AGK2 display an unfavorable plasma protein binding potential, which predicts a reduced bio-availability of these compounds. Furthermore, the bio-availability of sirtinol and AGK2 might be further limited due to the high lipophilicity (i.e., high miLogP value). Suramin is a very large molecule with very unfavorable drug-likeness parameters.
It is well accepted that sirtuins play an essential role in cancer cell proliferation and metabolism as well as in aging and inflammation processes . Therefore, modulating sirtuin activities could have therapeutic value. For instance, upregulation of SIRT1 has been reported in multiple cancer cell lines, indicating that SIRT1 inhibitor may be useful as therapeutic agents. Furthermore, non-sirtuin HDACi as well as SIRT2 inhibitors may be of interest to reduce neurodegenerative processes . Here, we identified that the sequiterpene alkaloid eurochevalierine displays a selective sirtuin 1/2 inhibitory profile, which provide a novel chemical scaffold for the lead compound development of SIRT1- and 2-based cancer therapy. Nonetheless, the type of inhibition (i.e., reversible or irreversible) and mechanism (e.g., competitive or noncompetitve) remains to be investigated.
Suramin, originally used for the treatment of trypanosomiasis and onchocerciasis, was described as a potent in vitro SIRTi (SIRT1, 2, and 5) . Accordingly, this symmetric polyanionic naphthylurea compound is frequently used as a positive control for in vitro sirtuin activity assays; however, to the best of our knowledge there is no report attesting to the capacity of suramin to trigger increased lysine acetylation of sirtuin targets. Conversely, we showed in this study that suramin failed to increase the acetylation level of two well characterized sirtuin targets (Figure S2). These results could be explained by the incapacity of suramin to reach in cellulo its cytoplasmic and/or nuclear sirtuin targets due to its relatively high molecular weight or by its inactivation through metabolic transformation. Such hypothesis is in agreement with the unfavorable predicted parameters for suramin drug-likeness (Table 4). In contrast, eurochevalierine was able to inhibit SIRT1 and 2 activities in vitro with IC50 values in the same range as suramin but also to induce the acetylation of a cytosolic (α-tubulin) as well as a nuclear target (histone H4) of SIRTs. Furthermore, eurochevalierine appears as a suitable drug candidate with more favorable parameters compared to the other tested reference SIRTi.
Sirtinol  as well as cambinol  described as selective SIRT1 and 2 inhibitors were previously described to act as potent anti-inflammatory agents. Further study suggested that the anti-cancer properties of sirtinol are linked to decreased proliferation capacities mediated by the development of a senescence-like growth arrest phenotype in human breast cancer MCF-7 and lung cancer H1299 cells . Remarkably, Peck et al. reported that the SIRT1 and 2 inhibitors sirtinol, nicotinamide, and salermide decrease proliferation of cancer cells without affecting cell cycle distribution as observed here with eurochevalierine (Figure 2A), whereas the SIRT1-specific inhibitor EX-527 triggers G1 cell cycle arrest . Similarly, AK-1, a specific SIRT2 inhibitor, induces G1 cell cycle arrest via the upregulation of p21 in colon HT29 cells [45,46]. Recently, it has been shown that EX-527 and AGK2 decrease Hela cervical cancer cell proliferation associated with G1 phase arrest and the downregulation of CDK4 and/or CDK6 . Moreover, effects leading to increased α-tubulin acetylation and modulation of microtubule dynamics and architecture could also contribute to impaired mitosis and proliferation . Altogether these results indicate that the inhibition of SIRT1 and SIRT2 activities by eurochevalierine may account for its cytostatic anti-cancer properties observed previously .
It should be noted that eurochevalierine also exhibited inhibitory effects against HDAC6 and 11 activities, albeit at higher concentrations. Considering the major role of HDAC6 in regulating many cellular processes and its implication in cancer progression, over the past few years, HDAC6 has emerged as an attractive pharmacological target, whose inhibition alone or in combination treatments displays promising anti-cancer properties in various cancer subtypes including multiple myeloma [8,48]. Thus, we cannot rule out the possibility that these combinational inhibitory effects may contribute to the selective anti-cancer properties of eurochevalierine.
4. Materials and Methods
Eurochevalierine was isolated from solid culture of Neosartorya pseudofischeri according to a procedure previously published . Eurochevalierine, nicotinamide (Sigma, Bornem, Belgium), suramin (Sigma), and SAHA (Cayman, Bio-connect, Huissen, The Netherlands) were dissolved in DMSO.
4.2. Cell Culture and Viability Assay
Cancer cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA), from the European Collection of Cell Culture (ECACC; Salisbury, UK), and from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ; Brauschweig, Germany). These cell lines include the human SK-MEL-28 melanoma (ATCC code HTB-72), the U-373 glioma (ECACC code 08061901), the A-549 non-small-cell lung cancer (DSMZ code ACC107), the K-562 chronic myeloid leukemia (DSMZ code ACC10), and the murine B16-F10 melanoma (ATCC code CRL-6475) models.
PBMCs were isolated as previously reported .
Primary CD34+ progenitor/stem cells were isolated from human umbilical cord blood obtained from the Clinique Bohler (Hôpitaux Robert Schuman, Luxembourg, Luxembourg) with the written informed consent of parents with the approval of the National Research Ethics Committee of Luxembourg. First, mononucleated cells were isolated by Ficoll™ (GE Healthcare, Roosendaal, The Netherlands) density gradient centrifugation. CD34+ cells were then purified out from mononucleated cells using magnetic cell sorting following the manufacturer’s instructions (MACS Miltenyi, Utrecht, The Netherlands).
High purity CD34+ cells were treated after 3 days of amplification in Stemline® II Hematopoietic Stem cell Expansion Medium (Sigma) supplemented with 1% antibiotic-antimycotic (BioWhittaker®, Lonza, Verviers, Belgium), 4 mM l-glutamine (BioWhittaker®), 50 ng/mL stem cell factor (ReliaTech, Wolfenbüttel, Germany), and 10 ng/mL interleukin 3 (Reliatech, Wolfenbüttel, Germany). Other cell models were cultured in RPMI1640 (BioWhittaker®) supplemented with 10% heat-inactivated fetal calf serum (BioWhittaker®) and 1% antibiotic-antimycotic. Cell lines were cultured at 37 °C in a humid atmosphere and at 5% CO2.
PBMCs and CD34+ cells were processed through a semi-automated image-based cell analyzer (Cedex XS Innovatis, Roche, Luxembourg), which provides information about cell concentration and viability based on the Trypan Blue exclusion method.
4.3. Flow Cytometry
Cells were collected, fixed, and stained with the APO AF TUNEL detection kit (BD, Erembodegem, Belgium) allowing analysis of both apoptosis and cell cycle at the same time . The experiment was conducted accordingly to the procedure of the manufacturer. Data were obtained through flow cytometry with a Cell Lab Quanta apparatus (Analis, Suarlee, Belgium).
4.4. In Vitro HDAC Activity Assay
HDAC assays were carried out as previously described .
4.5. Docking Studies
Docking studies were performed as previously described  with a docking grid size of 25 Å × 25 Å × 25 Å, which encompassed the entire inhibitor binding pocket of SIRT1 and 2 structures. To prepare the SIRT coordinates for docking simulation, ligands and all water molecules were removed except static water molecules in the binding site that could play important roles in the actual inhibition.
4.6. Protein Extraction and Western Blotting
Total proteins were extracted using the MPER® reagent (Thermo-scientific, Erembodegem-Aalst, Belgium) supplemented by 1× protease inhibitor cocktail (Complete EDTA-free, Roche, Prophac, Luxembourg, Luxembourg) according to manufacturer’s instructions. Histones were extracted as previously described . Western blots were carried out as previously reported .
Anti-acetylated α-tubulin (sc-23950) was from Santa Cruz Biotechnology (Tebubio, Boechout, Belgium); anti-acetylated histone H4 (06-866), anti-acetylated histone H3 lysine 56 (04-1135), anti-histone H1 (05-457) were from Millipore (Brussels, Belgium); anti-β-actin (A5441) was from Sigma. Corresponding secondary antibodies were from Santa Cruz Biotechnology.
4.7. Calculation of Drug-Like Properties
Statistical analyses were carried out using the GraphPad Prism 6.0 software (GraphPad Software, La Jolla, CA, USA). One-way ANOVA followed by the Holm–Sidak multiple comparison tests were used for statistical comparisons. p-values less than 0.05 were considered statistically significant.
The following are available online; Figure S1: Control docking results; Figure S2: Suramin fails to induce histone H4 or α-tubulin acetylation in K-562 cells; Table S1: Global drug-likeness parameters calculated for reference sirtuin inhibitors and eurochevalierine.
M.S. was supported by a “Waxweiler grant for cancer prevention research” from the Action Lions “Vaincre le Cancer.” This work was supported by Télévie Luxembourg, the «Recherche Cancer et Sang» foundation and the «Recherches Scientifiques Luxembourg» association. The authors thank the «Een Häerz fir Kriibskrank Kanner» association and the Action Lions “Vaincre le Cancer” for generous support. M.Die. and B.W.H. are supported by the Tumor Microenvironment GCRC (2011-0030001) from the National Research Foundation funded by the Ministry of Science and ICT of Korea. R.K. is a director of research with the Fonds National de la Recherche Scientifique (FNRS; Belgium).
M.S. and J.Y.J. designed and performed the experiments, analyzed the data, and wrote the paper; V.M. and F.L. performed the experiments; M.M., A.K., and A.E. provided the compound eurochevalierine; H.-J.K., R.K., M.Dic., B.W.H., and M.Die. conceived/discussed the experiments and wrote the paper.
Conflicts of Interest
The authors declare no conflict of interest.
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Figure 1. Structure of eurochevalierine.
Figure 2. Effect of eurochevalierine on cell cycle distribution. Cells were treated with 50 µM eurochevalierine for 72 h and then stained with propidium iodide (PI), and DNA content was measured by flow cytometry. (a) Cell cycle distribution profile in A-549 cells. Results of three independent experiments are represented on the same graph. (b) Quantification of the number of cells in sub-G1 fraction as a percentage of the total cell population. Results are expressed as mean ± SD of at least three independent experiments.
Figure 3. Eurochevalierine displays a moderate effect on in vitro non-sirtuin HDAC activities. (a) In vitro total HDAC activity was evaluated using K-562 nuclear extracts in the presence of various concentrations of eurochevalierine. (b) Selective in vitro HDAC activity assays were carried out using selected recombinant HDAC proteins in the presence of various concentrations of eurochevalierine. Results are reported as percent of DMSO vehicle control and expressed as the mean ± SD of three independent experiments. * and ** indicate p < 0.05 and p < 0.01 versus untreated cells, respectively.
Figure 4. Eurochevalierine docked into human SIRT1 and SIRT2. (a) Docking pose of eurochevalierine on the crystal structure of SIRT1 (bright orange; PDB code 4ZZI). The carboxamide derivative (light magenta), the inhibitor in the original SIRT1 crystal structure, was superposed. (b) Docking pose of eurochevalierine on the crystal structure of SIRT2 (green; PDB code 4RMH). SirReal2 (yellow), the inhibitor in the original SIRT2 crystal structure, was superposed. Close-up views on the right show that eurochevalierine (cyan) binds to the active site of the SIRT1 and 2 structures. The residues involved in hydrophobic effects with eurochevalierine are represented as gray-colored sticks. Static water molecules are shown as yellow dots.
Figure 5. Effect of eurochevalierine on the acetylation status of sirtuin targets. SK-MEL-28, U-373, and A-549 cells were treated with 5 and 50 µM eurochevalierine (EU) for 8 h. Acetylation of histone H4 (AcH4), histone H3 at lysine 56 (AcH3K56), and α-tubulin (Ac α-tubulin) was analyzed via Western blot. β-actin and histone H1 were used as loading controls for the analysis of total and acid extracts, respectively. SAHA (S, 1 µM) was used as a reference HDAC inhibitor. Blots are representative of three independent experiments.
Figure 6. Eurochevalierine does not alter the viability of normal cells. The viability of PBMCs from healthy donors was assessed after 24 and 48 h of treatment at the indicated concentration of eurochevalierine. Results correspond to mean ± SD of three independent experiments.
Figure 7. Only very high concentrations of eurochevalierine impair the growth of healthy primary human CD34+ stem/progenitor cells. CD34+ cells were incubated with the indicated concentrations of eurochevalierine and cell growth (a) and viability (b) were assessed after 72 h of treatment. Results correspond to the mean ± SD of three independent experiments with * and ** indicate p < 0.05 and p < 0.01 versus control.
Table 1. In vitro inhibitory activity of selected inhibitors and eurochevalierine on SIRT1, -2, and -3 activities.
|Compound||IC50 (µM) 1|
|Nicotinamide 2||97 ± 15||27 ± 3||67 ± 10|
|Suramin 3||2.8 ± 0.3||13 ± 1||>100 4|
|Sirtinol 3||82.5 ± 7.1||47.1 ± 4.0||ND|
|EX-527 3||0.10 ± 0.06||20.1 ± 4.2||ND|
|AGK2 3||98.1 ± 2.4||2.8 ± 1.0||ND|
|Eurochevalierine||9.8 ± 2.0||10.2 ± 3.9||>100 5|
1 IC50 values represent the mean ± SD of three independent experiments. ND: not determined. 2 Nicotinamide was use as in vitro reference SIRT3 inhibitor. 3 Suramin and sirtinol, EX-527, and AGK2 were used as in vitro reference inhibitors of SIRT1/2, SIRT1, and SIRT2, respectively. Data from  obtained in the same experimental conditions. 4 70% and 5 59% are the remaining enzymatic activity at 100 µM compared to the control set to 100%.
Table 2. Qualitative molecular docking results of eurochevalierine against human SIRT1 1.
1 Binding affinity energy values (kcal/mol) with SIRT1 proteins in the indicated protein data bank (PDB) codes. Sirtinol and EX-527 were used as reference inhibitors of SIRT1/2 and SIRT1, respectively.
Table 3. Qualitative molecular docking results of eurochevalierine against human SIRT2 1.
1 Binding affinity energy values (kcal/mol) with SIRT2 proteins in the indicated PDB codes. Sirtinol and AGK2 were used as reference inhibitors of SIRT1/2 and SIRT2, respectively.
Table 4. In silico drug-likeness parameters calculated for reference sirtuin inhibitors (SIRTi) and eurochevalierine.
|Rule of 5||n-atoms||20 ≤ x ≤ 70||38||86||9||30||17||30|
|MW (KDa)||180 ≤ x ≤ 500||526.63||1297.3||122.13||394.47||248.71||434.28|
|Absorption||BBBP||0.1 ≤ MA ≤ 2||0.10||0.04||0.34||3.10||4.10||0.08|
1 BBBP: blood-brain barrier penetration; IA: intestinal absorption; miLogP: octanol-water partition coefficient; MW: molecular weight; n-atoms: number of atoms; n-OHNH: number of hydrogen bond donors; n-ON: number of hydrogen acceptors; n-rotb: number of rotatable bonds; PPB: plasma protein binding; TPSA: topological polar surface area. EU: eurochevalierine; MA: middle absorption; NA: not applicable.
© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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The Algerian delegation, led by Mr. Rachid BLADEHANE, Secretary General of the Ministry of Foreign Affairs presented the Voluntary National Review on the implementation of the Sustainable Development Goals (SDGs) in Algeria, the Ministerial segment of the High Level Political Forum for sustainable development of the United Nations Economic and Social Council (ECOSOC).
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MANILA, Philippines - The Philippines will close its retail Treasury bonds (RTB) issue ahead of schedule and expects to raise a record P150 billion to P180 billion ($3.5 billion) as small investors become more confident with longer-term placements, Deputy Treasurer Eduardo Mendiola said yesterday.
“Total retail bond sales could easily reach P150 billion,” Mendiola said. The country had raised P110 billion at its last retail bond sale in October, which was a record at the time.
The response from retail investors shows they are no longer afraid of long-term investments, as a liquid secondary market in the paper offers them an exit route, he said over the telephone.
Total bond sales reached P134 billion as of Wednesday, including P49.6 billion sold at an auction on Tuesday and purchases by state firms of P16 billion, Mendiola said.
The Treasury was no longer keen on offering a second retail bond issue this year, he added.
The nine-day public offer for 5.375 percent 2027 bonds and 5.875 percent 2032 bonds was originally set to end on Feb. 29.
One of the government’s underwriters, First Metro Investment Corp. had earlier said Manila could easily top the P110 billion raised from October’s retail bond sale.
“We are looking at setting a new high here,” said First Metro Investment Corp (FMIC) president Juanchito Dispo.
FMIC is one of the issue managers tapped by the government for the RTB sale.
National Treasurer Roberto Tan said the government is very pleased with the strong demand. “ We are very pleased with the robust demand. The strong build up of volume shows deep appetite by investors for government securities that provide good returns and liquidity,’’ Tan said.
The Philippines relies heavily on local and foreign borrowing to fund its budget deficit, expected to hit 2.6 percent of GDP this year, or P279 billion year, from an estimated two percent in 2011.
Mendiola reiterated the retail bond offer was meant to provide small investors with safe investment options and should not alter the government’s local borrowing plan for the year.
Apart from First Metro Investment, BDO Capital, BPI Capital, and Deutsche Bank are issue managers of the bonds.Landbank of the Philippines, Development Bank of the Philippines, Philippine National Bank, China Banking Corp. are joint coordinators. - By Iris C. Gonzales