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Net Promoter Score® (NPS®) and service delivery styles
Transactional Net Promoter Score (NPS), where you ask customers to indicate their willingness to recommend your processes rather than their overall relationship with your organisation,
There are several options and tools for NPS, many that you may have already come across in everyday interactions with the companies that serve you. This includes surveys over the phone, survey machines in-store, and digital surveys sent via email and text.
Transactional NPS involves the collection of scores immediately after an interaction. It's particularly useful for gathering qualitative data directly tied to a particular stage of the customer journey. It also allows a business to react more quickly to customer feedback so that decisions for improvement can be made based on direct feedback.
There are pros and cons to each method. For example, a customer is less likely to give honest negative feedback at the end of a phone call to the same customer representative that conducted the call under review, so outsourcing the feedback call straight away to a call centre would be a better option, albeit more costly.
Sending out a survey alone to all your customers is not a hard task. But getting NPS to work is beyond just sending out the survey.
You'll need to send out a survey on a system that will automatically collect the results so that your team can spend their time actioning any negative feedback as quickly as possible instead of spending hours on manual input.
Automatically being alerted to negative feedback will also allow you to quickly close the loop before a bad review hits the public forum.
If you are looking to execute on a one time survey to test the benefit of an NPS feedback survey, then yes, you could try it out without buying into a system.
The process itself can be planned months in advance to ensure that data is collected properly if manual input is necessary. But if you are looking to benefit from transactional NPS, you could be sending out and reacting to thousands of personalised surveys every day. For that, you will definitely need end-to-end NPS tool which will automate the most resource-draining parts of the process.
Understanding the way tools work in NPS can only be achieved if you understand the different service types that require different tools.
'The Dynamics of Service' a book by Barbara Gutek talks about the two different types of customer delivery, one being a relationship service delivery and the other being an encounter delivery.
The former involves a longer-term relationship for the duration of the service while the other is as the name suggests, merely an encounter, much like the person serving you a meal at McDonald's.
Understanding that each type of service has different requirements and expectations is integral to a successful feedback process. An NPS tool such as Customer Gauge can differentiate between the two to ensure the right questions are being asked and the right kinds of data being collected.
Transactional Net Promoter Score (NPS), where you ask customers to indicate their willingness to recommend your processes rather than their overall relationship with your organisation,
With the expanding use of Transactional Net Promoter Score by a range of organisations, there is a growing need to collect large volumes of survey
You've read about it and convinced senior management that Net Promoter is the way to go. You know you will need a change management process to
Implementing a system that will automate the data collection and reporting elements of a Net Promoter Score (NPS®) process is an important step in integrating


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After surveying 1,125 people, the researchers found that 77% of those born between the early 1980s and mid-1990s rank ESG as their top priority when considering investment opportunities.
Anticipated returns, past performance, risk tolerance and other traditional factors are also important for investors, but were only cited by 10% or less of millennial respondents.
Nigel Green, CEO of the deVere Group, said: "Investments that score well in terms of ESG credentials often outperform the market and have lower volatility over the long run.
"For this reason, and because the biggest-ever generational transfer of wealth – likely to be around $30trn (£23trn) – from baby boomers to millennials will take place in the next couple of years, ESG investing is set to grow exponentially in the 2020s.
"As responsible investing becomes increasingly mainstream, we can also expect institutional investors, such as pension funds, to pile into ESG over the next few years."
The survey was carried out across Europe, North America, Australia, India, Asia and the Middle East.
It comes after the Thinking Ahead Institute found that assets managed in ESG mandates by the world's 500 largest fund managers rose by almost a quarter in 2018.
The findings show that ESG mandates grew by 23.3% in 2018, while total assets under management fell 3% from the previous year to $91.5trn.
The study also found that assets managed according to ESG principles grew 17.8% in 2018, while client interest in sustainable investing, including voting, rose 83%.
"Millennials appear to be leading the charge in socially responsible and impactful investing," Green continued.
"They understand that it is perfectly possible – and increasingly necessary – to make a profit while positively and proactively protecting people and the planet.
"These principles will fundamentally reshape the retail and institutional investment landscape in the next decade."
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2016 marks the 75th anniversary of Franklin D. Roosevelt's "Four Freedoms" speech. Franklin Roosevelt was elected president for an unprecedented third term in 1940 because at the time the world faced unprecedented danger, instability, and … [Read more...] about Franklin Roosevelt and the Four Freedoms Speech
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10 Ways to Lose Your Campaign
Some consultants will try and tell you they have a sure-fire way to win a campaign. There are no sure things in politics, but here are 10 things that will almost guarantee a loss on Election Day. Be warned: you still might win if your opposition is … [Read more...] about 10 Ways to Lose Your Campaign
Come on people, how hard is it to put "paid for by …" on something???
A lawyer friend who does a lot of campaign-related work emailed me this week with a list of her concerns about some candidates not understanding campaign laws/regulations. For experienced campaign workers, it was nothing complex or arcane, just … [Read more...] about Come on people, how hard is it to put "paid for by …" on something???
Letter Visibility for High Impact Signage
Yet here's a sign... Out, damned signs! Out, I say!" Anyone who has worked on campaigns for any period has a love-hate relationship with signs. I won't get into the pros and cons of sign today, but if your campaign is going to invest heavily in … [Read more...] about Letter Visibility for High Impact Signage
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*Prescription Required: Vetsource®, our pharmacy provider, may need to contact your veterinary hospital for prescription verification.
Bravecto® Topical for Cats kills adult fleas and treats and prevents flea infestations for 12 weeks. Bravecto® Topical for Cats also treats and controls black-legged tick infestations for 12 weeks and American dog tick infestations for 8 weeks.
Side Effects
Side effects may include vomiting, itching, diarrhea, hair loss, decreased appetite, lethargy, and scabs/ulcerated lesions.Other side effects may occur. If you notice anything unusual, contact your veterinarian.
Cats (at least 6 months of age and weighing at least 2.6lbs)
Bravecto® Topical for Cats is the only flea and tick treatment to last 12 weeks with one convenient topical dose.
Bravecto® Topical for Cats should be administered topically as a single dose as directed by your veterinarian.
Bravecto® Topical for Cats is not effective against American dog ticks beyond 8 weeks of dosing. For topical use only. Avoid contact with application site until dry. Avoid oral ingestion. The safety of Bravecto® Topical for Cats has not been established in breeding, pregnant and lactating cats. Use with caution in cats with a history of neurologic abnormalities. Neurologic abnormalities have been reported in cats receiving Bravecto® Topical for Cats, even in cats without a history of neurologic abnormalities.
Make sure to tell your veterinarian what other medication you are giving to your pet. Quite often your veterinarian may prescribe two different medications, even if a drug interaction may occur. In this case, your veterinarian may vary the dose and/or monitor your pet more closely. Contact your veterinarian if your pet experiences any unusual reactions when different medications are given together.
Allergic reactions to medications may occur. Be sure to inform Vetsource and your veterinarian if your pet has any known drug sensitivities or allergies. If your pet displays symptoms of an allergic reaction, call your veterinarian immediately or go to a veterinary emergency clinic. Symptoms may include but are not limited to: swollen lips, tongue, face, airways; difficulty breathing; agitation; profuse salivation; vomiting; widespread hives and itching.


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- Home /
- Cooking /
- Ranges /
- Gas: Freestanding /
- GE Profile 30" Free-Standing Gas Convection Range
4.5 32 Reviews
GE Profile 30" Free-Standing Gas Convection Range
Model# PGB911ZEJSS
Available Colors/Finishes
Product Inquiry Form
All fields marked * are required.
Quick Specs
Style: Gas: Freestanding
Width: 30 inches
Depth: 29 inches
Number Of Burners: 4
Ignition Type: ELECTRONIC
Continuous Grate: No
Self Clean: No
Dual Fuel: No
Down Draft: No
Convection Oven: Yes
Proof Mode: No
Warming Drawer: No
Storage Drawer: Yes
Burner Type: Sealed
Sabbath Mode: Yes
Edge-to-edge cooktop
A large cooking surface gives you extra room for pots and pans of all sizes and lets cookware move easily from burner to burner
Extra-large, integrated reversible cast iron grill/griddle
Enjoy ultimate cooking versatility. Cook up to six grilled cheeses on the industry's largest integrated griddle, and then flip it over to grill your favorite foods.
Two extra-large 18K Power Boil burners
Get twice the high-heat cooking power with two up-front extra-large burners
Gas convection oven
Circulates heat for high-quality results
Chef Connect
Three options with a single touch. For instant convenience, choose when to automatically sync clocks and the range burners with the microwave's surface light and vent.
3 oven racks
Enjoy extra oven space with three adjustable racks
Center oval burner
Large oval fifth burner provides ideal coverage for cooking with the reversible grill/griddle
Self-clean with Steam Clean option
Clean your oven the way you want
Heavy duty, dishwasher safe grates
Easily and safely clean grates in the dishwasher or place in the bottom of the self-clean oven
Black gloss oven interior
Enjoy a clean and sleek appearance
5.6 cu. ft. oven capacity
Enough room to cook an entire meal at once
|Total Capacity (cubic feet)
|App Filter - Cleaning Type
|Steam and Self-Clean
|App Filter - Control Type
|Electronic touchpad
|App Filter - Drawer Type
|App Filter - Cooktop Type
|Continuous Grates with Griddle
|App Filter - Cooking Technology
|App Filter - Cabinet Width
|Cooking 30 in. (Standard)
|App Filter - Product Type
|Ranges Free-Standing Single Oven
|App Filter - Convection
|App Filter - Fuel Type
|App Filter - Edge-to-Edge Cooktop
|Control Knobs
|Stainless Steel Appearance
|Handle Color
|Stainless Steel
|Handle Design
|Designer Style
|Cooktop Burner Grate Color
|Matte Black
|Oven Door Features
|Big View Oven Window
|Color Appearance
|Stainless Steel/Gray
|Top Burner BTU Rating - LP (000's BTU's)
|(1) 18.0; (1) 5.0; (1) 18.0; (1) 9.5; (1) 8.0
|Bake/Broiler BTU Rating - Nat. Gas (000's BTU's)
|16.0 Bake / 13.5 Broiler
|Top Burner BTU Rating - Nat. Gas (000's BTU's)
|(2) 18.0; (1) 10.0; (1) 9.5; (1) 5.0
|120v; 60Hz; 15A
|Bake/Broiler BTU Rating - LP (000's BTU's)
|16.0 Bake / 12.0 Broiler
|Warranty Notes
|See written warranty for full details
|Parts Warranty
|Limited 1-year entire appliance
|Labor Warranty
|Limited 1-year entire appliance
|Griddle Accessory
|Reversible Integrated Grill/Griddle (Included)
|Reversible Integrated Grill/Griddle (Included)
|LP Conversion Kit
|Center Grate
|Net Weight
|Height to Cooking Surface (in.)
|36 - 36 1/2
|Depth Without Handle
|Overall Depth
|Depth with Door Open
|Oven Interior Dimensions (W x H x D) (in.)
|24 x 19-1/2 x 20-5/8
|Overall Width
|Overall Height
|Approximate Shipping Weight
|Cabinet Width
|Burner - Right Front
|18,000 BTU Power Boil
|Burner - Left Rear
|5,000 BTU Precise Simmer
|Oven Cleaning Type
|Self-Clean with Steam Clean Option
|Cooktop Burner Type
|Burner - Right Rear
|Oven Rack Features
|3 Oven Racks
|Fuel Type
|Natural Gas (factory set)
|Burner - Center
|10,000 BTU Oval
|Cooking Technology
|Cooktop Surface
One-Piece Upswept
Stainless Steel
|Range with Storage Drawer
|Burner - Left Front
|18,000 BTU Power Boil
|Cooktop Burner Grate Material
|Heavy Cast Iron
|Control Type
|Electronic Touch
|Oven Cooking Modes
|Convection Bake
Convection Roast
|Ignition System
|Oven Control Features
|Audible Preheat Signal
Auto Self Clean
Certified Sabbath Mode
Chef Connect
|Product Type
|Free-Standing Single Oven
|Drawer Type
|Interior Oven Light
|1 Auto Incandescent
|Cooktop Type
|Continuous Grates with Griddle
|Gas: Freestanding
Product Downloads
|Installation Instructions
|Click here to download
|LP Conversion Kit
|Click here to download
|Click here to download
|Quick Specs
|Click here to download
No rebates currently available for this model.
out of 5 (32 Reviews)
Slick and exceptional performance
by: Anonymous
Yes, I would recommend this to a friend.
Source: ge profile.com
Submitted: 11-29-2018
It actually works really well.
by: Anonymous
Yes, I would recommend this to a friend.
Source: ge profile.com
Submitted: 11-15-2018
Great Stove
by: Anonymous
No, I would not recommend this to a friend.
Source: ge profile.com
Submitted: 10-30-2018
Great features
by: Anonymous
Yes, I would recommend this to a friend.
Source: ge profile.com
Submitted: 10-04-2018
Love all the features
by: Anonymous
Yes, I would recommend this to a friend.
Source: ge profile.com
Submitted: 09-30-2018
Streamline your kitchen's look with the modern styling from GE Profile appliances. This sleek line of appliances is as innovative as it is beautiful, featured in lustrous finishes like the bold new black stainless steel, smooth stainless steel, and the revolutionary smudge-proof Slate™ and Black Slate™. Customize the look of your kitchen with integrated appliances such as the counter-depth refrigerators featuring showcase LED lighting.
- Slate - A warm matte alternative to stainless steel that is both smudge-proof and magnetic.
- Black Slate - A deep charcoal take on stainless steel featuring a matte finish to hide fingerprints and smudges.
- Stainless Steel - A classic, neutral tone designed for every kitchen style.
- Black Stainless Steel - A sleek bold statement featuring brushed metal stainless steel finish and a glossy black hue.
Innovative Features
- Keurig ® K-Cup Brewing System - The advanced water filtration system gives you cleaner tasting water, even the water you brew your next coffee at your GE Café™ refrigerator with the Keurig® K-Cup® Brewing System
- Advantium Technology - The True European Convection with Direct Air found in GE Profile wall ovens and over-the-counter microwaves that feature GE Advantium technology will quickly and evenly bake your dishes to perfection every time
- WiFi Connect - From monitoring your refrigerator to adjusting your oven's temperature, you can control your kitchen with Wi-Fi connected appliances featured in GE Profile ranges, wall ovens, dishwashers, washers, and dryers.
- Hands-free French Door Autofill - Now you can schedule your hot water with Hot Water Scheduling then let Hands-Free Autofill fill your container for you.
- Bottle-Wash Jets - Enjoy the ultimate clean with over 140 cleaning jets found in GE Profile dishwashers, including dedicated bottle-wash and deep-clean silverware jets, exclusive wash zones for true half loads, and a versatile third rack for additional space and flexibility. .


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What is PostKnife?
PostKnife is a subscription to have professionally sharpened knives, delivered to your door on a routine basis (commonly referred to as a 'knife service'). Two sets of knives are continuously being exchanged. On Knife Day your freshly sharpened knives arrive in the mail. Remove the sharp knives them from the box, then put the used knives into that same box. Adhere the enclosed return shipping label, and put the box out in the next day's mail.
How can I contact PostKnife?
Call us at 1 (608) 285-2623 -or- email us at email@example.com
If questions or concerns arise, please contact us. We're here to help.
Can I choose a custom set of knives?
Yes. Contact us to get started, and a PostKnife expert will help you choose a custom set.
How does PostKnife shape an edge?
At PostKnife, we sharpen a lot of knives, and need to be able to shape a professional edge efficiently. Our process is water cooled to retain the temper of the steel. If the temper is lost, the integrity is lost, and no matter how well the edge is shaped, it will not be durable. The first and most aggressive step in our sharpening process is called 'hollow grinding'. Hollow grinding is a thinning process that simultaneously removes steel from both sides of a blade. Hollow grinding requires skill to ensure that material is removed consistently and at an adequate rate along the length of the blade. After a blade has been hollow ground to the proper thickness, it is ready to be edged.
The 'edging' process at PostKnife is achieved by free hand flat grinding on a stand up grindstone. This is the traditional method of edging that has been used by my family for four generations. A stand-up grind stone can efficiently shape a precision edge on the vast majority of knives and blades, however, grinding with this equipment is a skill, and the operator must be experienced to use it effectively.
Once the edge has been shaped, we move on to buffing on a 220 grit buffing wheel. Buffing removes some of the deeper grinding marks/scratches, helps de-burr the edge, and contributes to shaping a finer edge overall.
The final step in our process is honing. Some knives are honed by hand on a flat stone while others can be finished in a honing machine. Honing is the least aggressive step in the process with stones at 440 grit. The burr which is created by shaping the edge, is completely removed while honing, and the result is a durable yet fine, razor sharp edge.
Can you sharpen my personal knives?
Yes. In fact, if you're signed up as a customer with PostKnife, we'll sharpen up to 2 personal knives per exchange at no additional charge. If you're not a PostKnife customer, click here to visit our sister company; they can help you out.
What happens if I damage or lose a knife?
We understand that accidents happen. If lost or damaged knives becomes a problem, we'll contact you.
What should I do if I can't find all of the used knives to return?
Don't worry, just return all of the old PostKnife knives you can find. When the others turn up, send them back in a future shipment.
When should I send back the used set of knives?
The knife service works best when you swap out the knives as soon as you receive a fresh set (fresh knives out of the box, used knives into that same box). The next day, put the box with your outgoing mail, and your mail carrier will pickup the box for return.
When do I get charged for the service?
The knife rental service works on a subscription format. You are charged on a recurring basis, once every two weeks. Contact us if you're interested in receiving service on a different schedule.
What happens if I forget to return the used set?
If you do not return the used set of knives, your subscription will still renew and you will be charged the regular service fee on your scheduled date, but we will not ship out any fresh knives until we receive the used set.
How much does it cost to start the knife service?
The cost of your first shipment is all you have to pay to get started. There are no additional startup costs, and you can cancel at any time.
Is shipping included?
Yes! All shipping is included in the service.
What shipping service does PostKnife use?
PostKnife ships via the US Postal Service (USPS). The shipping address that you provide, must be a physical address or PO Box that USPS delivers to. Please contact us if USPS delivery is not an option for you.
I love my knives. Why would I use PostKnife?
We love knives just like you do; properly maintaining and restoring knives is our passion. The PostKnife subscription provides your kitchen with a full set of sharp knives for everyone to use. That way, you can rest assured that sharp knives are available for your entire crew, and they can focus on the food. Sharpening knives professionally is all that we do; we'd love the opportunity to show you our work.
I've never heard of a knife service. Is this for real?
Yes! We have deep family roots to Trento, Italy, a region that is globally recognized as the birthplace of the knife grinding trade. For generations, delivering sharp knives to restaurant kitchens, chefs and meat cutters, has been our family's livelihood. Thousands of restaurants utilize a knife service to always have sharp knives.
Are these your knives or my knives?
The knives belong to PostKnife. Only knives owned by PostKnife are used in the service.
Do you carry any other types of knives?
Yes. We carry many different styles of knives. Contact us if you would like to choose your own custom set.
What is the phone number for PostKnife customer service?
1 (608) 285-2623
What is the mailing address for PostKnife?
S11251 Fairview Rd., Spring Green, WI 53588
If my business is not a restaurant, can I still get the PostKnife service?
Yes. PostKnife services a wide range of establishments including restaurants, butcher shops, grocery stores, bakeries, caterers, cafés, institutional kitchens, sandwich shops, dining hall kitchens, hotels, taco trucks, pizzerias and more. If you need sharp knives all the time, we're here to serve you.
How do I cancel service?
Easy. Just log into your account profile and under the service options tab choose to 'cancel'. When you cancel the knife service we can ship you a box and prepaid return shipping label, so you can easily return the PostKnife knives.
If the knives are not returned, PostKnife does reserve the right to charge you for the knives, and for shipping costs incurred.
What happens if I cancel service, but don't send the knives back?
All knives are owned by PostKnife, so you are responsible for returning them. If you would like to keep the knives, we'll be happy to sell them to you.
What kind of knives do you use?
All knives are PostKnife brand, manufactured with high quality stainless steel blades and poly handles.
Can I get knives delivered more or less often than every two weeks?
Absolutely. PostKnife offers weekly, biweekly and triweekly (once every 3 weeks) delivery service as well. Just contact us to customize your delivery schedule.
Is it really cheaper to use PostKnife than to buy knives and sharpen them in house?
Yes. Knife service costs less than continually buying knives, trying to maintain them in house, and unfortunately using dull knives much of the time. The value that PostKnife adds to a kitchen greatly exceeds the cost.
Safety - preventing one accident can pay for a year of subscription service. Take into consideration that an injury with a dull knife is usually much worse than a cut with a sharp knife. Loss in production occurs when an injured staff member has to stop working and sometimes go to the hospital. Often times an additional employee has to take the injured employee to the hospital, so that's 2 employees down! On top of that the injured employee is often needs a day or more off of work for recovery. At the end of the day a hospital bill or a workers comp claim will also be in order. The costs add up quickly. A dull knife is a dangerous knife.
Efficiency – reduce down time and food waste directly related to dull knives and trying to sharpen them. Doing prep work takes less time with sharp knives. Food orders can go out more efficiently; during a rush that translates into shorter wait times and more customers being served.
Consistency - with PostKnife you know that every knife in the kitchen is sharp all the time; that piece of mind makes staffing easier and enables your crew to focus on the food!
Other Knife Costs – NEVER BUY KNIVES AGAIN; PostKnife eliminates all knife purchase costs, and costs of sharpening stones, machines and gadgets.


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balance, harmony & nature
SAVA products were created to provide a solution for those who would like to nourish themselves with all natural and organic products. We use the best quality ingredients including Pure Essential Oils. All of our products are hand made in Switzerland with love and are created to nourish & revitalize your body, mind and soul.
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Consign Your Best Items with Lelands. We Also Pay Cash on the Spot! Learn More Here.
On July 24, 1934 at the cavernous Baker Bowl in Philadelphia, the Phillies defeated the Pittsburgh Pirates 9-0. Bucs first baseman Gus Suhr hit a foul ball which is was signed by 13 members of the Pirates including Hall of Famers Arkie Vaughan, Waite Hoyt, Pie Traynor and coach Honus Wagner. The signatures are written in black ink and all rate at least an "8" out of a possible 10. "Cookie' Lavagetto signed on the sweet spot. The ball is marked with the date and final score as well as the words, "Foul Ball By Suhr". This lot comes with LOA.
Other past auction items that may be of interest to you.
May 2002
December 2002


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Each year, Tennis Resorts Online publishes its rankings of the best tennis resorts and camps world wide. We based this year's list on reviews submitted between May 1, 2018 and April 30, 2019 by tennis vacationers like yourselves, and we then used that data to determine the rankings. There are a number of prestigious tennis resorts and tennis academies around the world offering the best tennis camps for adult players at all levels. John Newcombe Tennis Ranch, Texas; Saddlebrook Tennis Resort, Florida; Cliff Drysdale Tennis: Amelia Island Florida, La Costa California and Las Palmas California. Jonathan Markson Tennis.
Nike Adult Tennis Camps Immerse yourself thoroughly in tennis! Nike Adult Tennis Camps offer specific tennis improvement programs for beginning, intermediate and advanced levels.. Learn from America's finest coaches; Intensive tennis instruction, drills and match play. Adaptive tennis is about accommodating the way tennis is played and taught for the unique needs of players with differing abilities. Tennis is a sport for everyone. Adaptive tennis emphasizes inclusivity by providing everyone, including those with disabilities, the opportunity to play, have fun and fall in love with our lifetime sport.
Tennis enthusiasts of all levels flock to Saddlebrook's Tampa tennis camps for adults to experience our intense and demanding program.Location: 5700 Saddlebrook Way, Wesley Chapel, 33543, FL. Tennis camps are an awesome way to get fit while on holiday and travel to exotic destinations around the world at the same time. Designed to enhance your performance on the court, sharpen your skills, or simply to learn the basic rules of the game, adult tennis camps are bound to help you shed stress and meet new friends.
10 rows · As with the Adult Tennis Clinics, our Advanced Adult Tennis Clinics are offered in a . Located just north of Chicago, in Evanston, IL, Northwestern Tennis Camps offers year-round tennis programs. Our junior and adult tennis program includes classes for kids 4+ and adults of all abilities. Our summer tennis camps for kids 4-18 years old have been rated amongst the best on the North Shore.


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Area 86 is a smooth physics-based puzzle and exploration game set in space. Control your robot in an interactive environment. Find, combine, activate, pick up and throw different objects to solve puzzles.
Only registered members can share their thoughts. So come on! Join the community today (totally free - or sign in with your social account on the right) and join in the conversation.
Area 86 is a physics-based escape room style game set in space.
Find, pick, throw, activate & destroy items to escape.


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Title: Water harvesting from Soils by Solar-to-Heat Induced Evaporation and Capillary Water Migration
(Submitted on 22 Nov 2019)
Abstract: Fresh water scarcity is one of the critical challenges for global sustainable development. Several novel water resources such as passive seawater solar desalination and atmospheric water harvesting have made some progress in recent years. However, no investigation has referred to harvesting water from shallow subsurface soils, which are potential huge water reservoirs. Here, we introduce a method of solar-driven water harvesting from soils, which can provide cheap fresh water in impoverished, arid and decentralized areas. The concentrated solar energy is used to heat the soils to evaporate the soil moisture. Then vapors flow to the condenser through tubes and condense as freshwater. Sustainable water harvesting is realized by water migration due to capillary pumping effect within soils. In the laboratory condition, an experimental setup is designed and its water-harvesting ability from soils is investigated. The maximum water mass harvesting rate was 99.8 g h-1. In about 12 h, the total harvesting water could be as high as about 900 ml. The whole process is solar-driven and spontaneous without other mechanical or electrical ancillaries. The water harvesting rate under one sun energy flux (1 kW m-2) is estimated to be about 360 g h-1 with a 1 m2 solar concentrator. Our proposal provides a potential onsite and sustainable fresh water supply solution to deal with the water scarcity problem.
Submission historyFrom: Xiaotian Li [view email]
[v1] Fri, 22 Nov 2019 03:03:53 GMT (926kb)
Link back to: arXiv, form interface, contact.


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Home » Artwork » Manga and Super Deformed » Mirasawa Kuno
Mirasawa Kuno
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Kuno was the character of one of the players in a Magic Knight Rayearth fan roleplay we once did.
Kuno is a 14-year old schoolboy. After he gets teleported to the magical land of Cephiro, he develops magical powers and commands the element of fire to do his bidding as the first male magical girl of all times...
As his element is fire he should actually wear red pants but the player opted against that.
This too is from my very first days of digital colorizing so I don't have a better version to offer.
Friday, 20 December 2013
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Antioxidant and Anti-Osteoporotic Activities of Aromatic Compounds and Sterols from Hericium erinaceum
Korean Medicine (KM) Application Center, Korea Institute of Oriental Medicine, Daegu 41062, Korea
Department of Food and Nutrition, Hannam University, Daejeon 34430, Korea
College of Pharmacy, Chungnam National University, Daejeon 34134, Korea
Authors to whom correspondence should be addressed.
Academic Editor: Derek J. McPhee
Molecules 2017, 22(1), 108; https://doi.org/10.3390/molecules22010108
Received: 14 November 2016 / Revised: 2 January 2017 / Accepted: 2 January 2017 / Published: 11 January 2017
(This article belongs to the Section Natural Products Chemistry)
Hericium erinaceum, commonly called lion's mane mushroom, is a traditional edible mushroom widely used in culinary applications and herbal medicines in East Asian countries. In this study, a new sterol, cerevisterol 6-cinnamate (6), was isolated from the fruiting bodies of H. erinaceum together with five aromatic compounds 1–5 and five sterols 7–11. The chemical structures of these compounds were elucidated using chemical and physical methods and comparison of HRESIMS, 1D-NMR (1H, 13C, and DEPT) and 2D-NMR (COSY, HMQC, HMBC, and NOESY) spectra with previously reported data. The antioxidant and anti-osteoporotic activities of extracts and the isolated compounds 1–11 were investigated. All compounds exhibited peroxyl radical-scavenging capacity but only compounds 1, 3, and 4 showed potent reducing capacity. Moreover, compounds 1, 2, 4, and 5 showed moderate effects on cellular antioxidant activity and inhibited the receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastic differentiation. These results suggested that H. erinaceum could be utilized in the development of natural antioxidant and anti-osteoporotic nutraceuticals and functional foods.
Keywords: Hericium erinaceum; hericiaceae; antioxidant; anti-osteoporosis; sterol; aromatic compound
1. Introduction
Oxidative stress is caused by an imbalance in the generation of reactive oxygen species (ROS) and the activity of antioxidant defenses. Severe oxidative stress has been implicated in many chronic and degenerative diseases, including osteoporosis, cancer, aging, and neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis [1]. Several natural antioxidants have been reported, including flavonoids, coumarins, anthocyanins, and phenolic acids. Some natural flavonoids and coumarins with antioxidant activity, including scopoletin and baicalein, have been found to exert potent anti-osteoporotic effects through the suppression of osteoclast formation and tartrate-resistant acid phosphatase (TRAP) [2,3,4]. Therefore, interest in novel natural antioxidants has increased. Osteoclasts are tissue-specific macrophage polykaryons formed by the differentiation of monocyte/macrophage precursor cells at or near the bone surface in response to osteoclastogenic factors, particularly the receptor activator of nuclear factor κB ligand (RANKL), which performs important functions in osteoclast differentiation, including mediation and activation [5,6]. This developmental process results in TRAP expression in the differentiating cells, followed by their fusion to produce multinucleated cells capable of resorbing a mineralized matrix [7].
In our continuing search for antioxidant and anti-osteoporotic components from natural sources, we found that the MeOH extract of Hericium erinaceum showed potent effects. H. erinaceum, commonly called lion's mane mushroom, is an edible medicinal fungus found in China, Korea, and Japan, and is used as a raw material in a wide range of East Asian foods. Its fruiting bodies are used to treat gastric ulcers, chronic gastritis, and other digestive tract-related diseases [8]. The constituents of H. erinaceum include polysaccharides, heteropolysaccharides, diterpenoids, and aromatic compounds [9]. Previous pharmacological studies on compounds isolated from H. erinaceum also showed their stimulatory effects on nerve growth factor synthesis and antitumor activity [10,11]. However, no studies have investigated the active components of H. erinaceum with antioxidant and anti-osteoporotic activities. In this study, bioassay-guided isolation of the CHCl3 fraction of the MeOH extract of H. erinaceum yielded 11 compounds 1–11, which were evaluated for antioxidant activity using oxygen radical absorbance capacity (ORAC), cupric ion reducing antioxidant capacity (CUPRAC), and cellular antioxidant capacity (CAC) assays. Moreover, the anti-osteoporotic activities of these compounds were evaluated by investigating their inhibitory effects on osteoclastic differentiation.
2. Results and Discussion
Using combined chromatographic separations, 11 compounds, including five aromatic compounds 1–5 and six sterols 6–11, one of which is new (cerevisterol 6-cinnamate, compound 6), were isolated from the CHCl3 fraction of the MeOH extract of H. erinaceum fruiting bodies (Figure 1). The known compounds were identified as hericenone J (1) [12], N-dephenylethyl isohericerin (2), 4-[3′,7′-dimethyl-2′,6′-octadienyl]-2-formyl-3-hydroxy-5-methyoxy-benzylalcohol (3) [13], isohericerin (4) [14], hericerin (5) [14], (22E,24R)-ergosta-7,22-diene-3β,5α,6β,9α-tetraol (7) [15], cerevisterol (8) [16], blazein (9) [16], 3β,5α-dihydroxy-6β-acetoxy-ergosta-7,22-diene (10) [17], and ergosta-7,22-diene-3β,5α,6α-triol (11) [18]. The purity of all isolated compounds was confirmed to be >95% using HPLC. Compound structures were elucidated by comparison of spectroscopic data with reported values. Six compounds (6–11) were isolated from H. erinaceum for the first time.
Compound 6 was obtained as a yellowish oil. The molecular formula C37H52O4 was assigned based on a pseudomolecular ion peak at m/z 561.3944 [M + H]+ (calcd. 561.3938) in the HR-ESI-MS spectrum. The 1H-NMR spectrum of 6 (Table 1) contained peaks corresponding to six methyl groups [δH 0.53 (s, Me-18), 0.76 (d, J = 6.5 Hz, Me-26, 27), 0.84 (d, J = 6.7 Hz, Me-28), 0.95 (d, J = 6.5 Hz, Me-21), and 1.08 (s, Me-19)], two oxymethines [δH 4.02 (m, H-3α) and 4.93 (d, J = 5.4 Hz, H-6α)], and three olefinic protons at δH 5.09 (dd, J = 15.3, 7.7 Hz, H-22), 5.14 (dd, J = 15.3, 7.0 Hz, H-23), and 5.26 (d, J = 5.4 Hz, H-7). Additionally, cinnamic acid residue signals were observed at δH 6.33 (d, J = 16.0 Hz, H-2′), 7.30-7.31 (m, H-6′, 7′, 8′), 7.46 (d, J = 8.0 Hz, H-5′, 9′), and 7.58 (d, J = 16.0 Hz, H-3′). The 13C-NMR spectrum (Table 1) contained 28 resonances corresponding to the sterol moiety, which were assigned to six methyl groups (δC 12.5, 17.6, 18.5, 19.7, 20.3, and 21.2), seven methylene groups (δC 22.2, 22.8, 27.9, 30.7, 32.6, 39.3, and 39.4), eight sp3 methine groups (δC 33.1, 40.4, 43.4, 55.0, 56.0, 67.5, and 73.5), three sp2 methine groups (δC 114.3, 132.2, and 135.4), and four quaternary carbon atoms (δC 37.3, 43.9, 75.5, and 145.9). Signals from the cinnamic acid residue revealed a carbonyl group [δC 166.4 (C-1′)], two olefinic carbon atoms [δC 118.4 (C-2′) and 145.1 (C-3′)] and six aromatic carbon atoms [δC 128.2 (C-5′, 9′), 128.9 (C-6′, 8′), 130.4 (C-7′), and 134.4 (C-4′)]. The above data showed that the sterol moiety 6 was a Δ7-ergostane derivative, which was similar to compound 8 (cerevisterol) [19], but had a cinnamic acid residue attached at C-6. This finding was supported by a key HMBC correlation between H-6α (δH 4.93) and C-1′ (δC 166.4) (Figure 2). Alkaline hydrolysis of 6 with 0.5% NaOH yielded cerevisterol and cinnamic acid, which were identified by TLC analysis (CHCl3–acetone: 2:1; Rf = 0.35, 0.80) with known standards (see Supplementary Materials).
The relative configuration of 6 was determined using a NOESY NMR experiment and empirical pyridine-induced deshielding effects. NOE correlations were observed between H-6α (δH 4.93) and Heq-4 (δH 1.69), H-3α (δH 4.02) and Heq-4 (δH 1.69), and Me-19 (δH 1.08) and Hax-4 (δH 1.87), which indicated that the two hydroxyl groups were in 3β,6β-orientations (Figure 2) [18,19]. The chemical shifts of Hax-3, Hax-4, H-6, and Me-19 measured in pyridine-d5 were shifted downfield, which indicated the presence of a 3β,5α,6β-trihydroxylated system in compound 6 [15,20]. A comparison of the chemical shifts of C-26, C-27, and C-28 of 6 with those of compound 8 established a 24R configuration in 6 [21]. The large coupling constant (15.3 Hz) between H-22 and H-23 was indicative of an E configuration for the double bond [22]. Therefore, compound 6 was identified as cerevisterol 6-cinnamate [(22E,24R)-ergosta-7,22-dien-3β,5α,6β-triol 6-cinnamate].
The MeOH extract and CHCl3 and water fractions of H. erinaceum were evaluated for their antioxidant activity at concentrations of 1.0 and 10.0 μg/mL. The MeOH extract and CHCl3 fraction showed potent effects at 10.0 μg/mL in the ORAC and CUPRAC assays (Figure 3A,C). In order to identify the active antioxidant and anti-osteoporotic components of H. erinaceum, 11 compounds (1–11) from the CHCl3 fraction were evaluated using ORAC, CUPRAC, CAC, and TRAP assays. The antioxidant activities of compounds 1–11 were investigated by ORAC and CUPRAC assays. The ORAC assay is based on hydrogen atom transfer while the CUPRAC reducing capacity assay depends on single electron transfer [23]. The results showed that compounds 1, 3, and 6 had potent peroxyl radical-scavenging activities at 10 μM with ORAC values of 6.05, 4.98, and 8.01, respectively. Other compounds showed negligible activities with ORAC values below 4.00 (Figure 3B). In structure-activity analysis of the isolated compounds, the aromatic compounds 1 and 3 showed more potent activity than the other aromatic compounds (2, 4, and 5). The same hydroxyl group is present in the aromatic ring but compounds 2, 4, and 5 also contain a nitrogen atom. Therefore, it appears that the nitrogen atom in the aromatic ring exerts a negative influence on peroxyl radical-scavenging activity. Compound 6 exhibited the strongest activity among the isolated sterols (6–11). Based on structural analysis, the cinnamic acid residue at C-6 seems to be an important element for peroxyl radical-scavenging activity.
The ability to transfer single electrons was examined by reducing capacity assay. The reducing capacity of all compounds was evaluated based on the production of Cu(I) ions from Cu(II) ions in the presence of each isolated compound. The results revealed that only compounds 1 and 3 exhibited an obvious reducing capacity at 10.0 μM, with CUPRAC values of 6.16 and 5.16, respectively (Figure 3D). These results indicated that the ability of compounds 1 and 3 to donate hydrogen atoms or electrons to peroxyl radicals, and to convert them into relatively stable compounds, may contribute to their peroxyl radical-scavenging capacity.
The intracellular antioxidant capacities of compounds 1–11 were investigated by a CAC assay. First, cell viability was tested by an MTT assay. The results showed that compounds 1–11 were not significantly cytotoxic to HepG2 cells at the tested concentrations. Following treatment with AAPH, intracellular oxidative stress in HepG2 cells increased to 213.8% of the control value. As shown in Figure 4, all 11 compounds decreased the intracellular oxidative stress caused by AAPH to 111.6%–177.7% of the control value. Thus, all the isolated compounds may permeate the cell membrane and suppress the intracellular oxidative stress induced by peroxyl radicals.
The MeOH extract and CHCl3 fraction also exhibited moderate TRAP activity at 10.0 μg/mL (Figure 5A). We also investigated the anti-osteoporotic activity of compounds 1–11 by evaluating their inhibitory effects on TRAP activity in osteoclasts differentiated from RAW 264.7 cells. Osteoclast differentiation was induced by RANKL, a key cytokine regulating osteoclastogenesis and bone resorption. RANKL treatment strongly induced osteoclast formation from RAW 264.7 pre-osteoclasts and markedly increased TRAP activity [24]. TRAP is highly expressed in osteoclasts and is widely used as a phenotypic marker of osteoclasts. Based on the reduction in TRAP activity, the inhibitory activities of compounds 1–11 against osteoporosis are shown in Figure 5. At a concentration of 10 μM, compounds 8–11 showed strong inhibition of RANKL-induced TRAP activity, reducing it to 28.1%, 74.8%, 81.8%, and 138.9%, respectively, from a value of 332% in RANKL-treated control cells. The potent inhibitory effects of compounds 8–11 may be due to their strong cytotoxicity. In contrast, compounds 1, 2, 4, and 5 showed moderate inhibition at 10 μM (Figure 5B), which indicated that the presence of an α,β-unsaturated carbonyl moiety in the structure may be involved in the inhibitory effects on the TRAP activity of osteoclasts.
Recent studies have suggested that intracellular antioxidant activity related to ROS scavenging may be responsible for increased osteoblast differentiation in MC3T3-E1 pre-osteoblastic cells and suppression of the differentiation of RAW 264.7 pre-osteoclast cells into osteoclasts [2,25]. However, a comparison of the anti-osteoporotic and antioxidant activities of compounds 1–11 indicated their anti-osteoporotic activity appeared to be associated with cellular antioxidant activity instead of in vitro antioxidant activity, but was not proportional to cellular antioxidant activity. These results can be explained by the different plausible mechanisms that may contribute to cellular antioxidant activities, such as the induction of phase II detoxifying and antioxidant enzymes as an indirect source of antioxidant activity. In addition, the important role of the α,β-unsaturated carbonyl moiety in the indirect cellular antioxidant activity of these compounds (1, 2, 4, and 5) can be ignored. However, more research is required to determine whether the potent anti-osteoporotic activities of compounds 1, 2, 4, and 5 result from indirect antioxidant activity.
3. Materials and Methods
3.1. General Information
Optical rotations were determined using a DIP-370 automatic polarimeter (Jasco, Easton, MD, USA). The FT-IR spectra were measured using a Jasco Report-100 infrared spectrometer; The NMR spectra were recorded using an ECA 600 spectrometer (1H, 600 MHz; 13C, 150 MHz, JEOL, Tokyo, Japan). Mass spectra were recorded on an LCQ advantage trap mass spectrometer (Thermo Finnigan, San Jose, CA, USA) equipped with an electrospray ionization (ESI) source. High-resolution electrospray ionization mass spectra (HR-ESI-MS) were obtained using a 6530 Accurate-Mass Q-TOF LC/MS system (Agilent, Santa Clara, CA, USA). Column chromatography was performed using silica gel (Kieselgel 60, 70–230, and 230–400 mesh, Merck, Darmstadt, Germany) and YMC RP-18 resins (Merck), and thin layer chromatography (TLC) was performed using pre-coated silica-gel 60 F254 and RP-18 F254S plates (both 0.25 mm, Merck).
3.2. Fungal Material
Dried fruiting bodies of H. erinaceum were purchased from an herbal market in Kumsan, Chungnam Province, Korea in August 2013. The identity of the specimens was confirmed by one of authors (Young Ho Kim). A voucher specimen (CNU 13110) was deposited at the Herbarium of the College of Pharmacy at Chungnam National University.
3.3. Fermentation, Extraction and Isolation
Dried fruiting bodies (2.5 kg) of H. erinaceum were extracted with MeOH (5 L, × 3) under reflux. The MeOH extract (320.0 g) was suspended in water (5.0 L) and partitioned with CHCl3 (5.0 L) yielding CHCl3 (90.0 g) and water (220.0 g) fractions. The CHCl3 fraction (90.0 g) was subjected to silica gel (5.0 cm × 30 cm) column chromatography with a gradient of n-hexane–EtOAc–MeOH (25:1:0, 9:1:0, 5:1:0, 2.5:1:0, 1:1:0.1, 1:1:0.3, 0.5:1:0.5; 4 L for each step) to give eight fractions (Fr. 1A–1H).
Fraction 1C was separated on a silica gel column with a gradient of n-hexane–EtOAc (20:1 to 10:1, 10 L) to give 11 sub-fractions (Fr. 1C-1–1C-11). Fraction 1C-8 was subjected to YMC (1.0 × 80 cm) column chromatography eluted with a MeOH–acetone–H2O (3:3:1, 4:4:1, 6:6:1; 1.2 L for each step) gradient to give compounds 1 (43.6 mg) and 5 (86.2 mg). Fraction 1D was subjected to silica gel (2.5 cm × 30 cm) column chromatography with a gradient of n-hexane–EtOAc–MeOH (8:1:0.15, 6:1:0.15, 4:1:0.15, 3:1:0.15, 2:1:0.15, 1.5:1:0.15; 2.5 L for each step) to give 10 fractions (Fr. 1D-1–1D-10). Fraction 1D-7 was subjected to YMC (1.5 cm × 80 cm) column chromatography eluting with a MeOH–acetone–H2O (1.5:1:1, 3:1.5:1, 6:3.5:1, 9:5:1; 1.0 L for each step) gradient to give compound 3 (21.4 mg). Fraction 1D-8 was subjected to silica gel (1.0 cm × 80 cm) column chromatography with a n-hexane–EtOAc–acetone (3:1:0.15; 750 mL) mixture as eluent to give compound 4 (13.4 mg). Fraction 1D-9 was subjected to YMC (2.5 cm × 80 cm) column chromatography eluting with MeOH–acetone–H2O (1:1:1, 2:2:1, 3:3:1, 5:5:1, 8:8:1, 10:10:1; 3 L for each step) to give eight sub-fractions (Fr. 1D-9-A–1D-9-H). Fraction 1D-9-E was subjected to silica gel (1.5 cm × 80 cm) column chromatography with a CHCl3–acetone (7:1, 9:1, 12:1; 1 L for each step) eluent to give compounds 9 (12.0 mg), 10 (7.0 mg), and 11 (22.0 mg). 1D-10 was subjected to YMC (2.5 cm × 80 cm) column chromatography eluting with MeOH–H2O (1:1, 2:1, 3:1, 5:1, 7:1, 9:1, 11:1, 13:1, 15:1; 2.5 L for each step) to give seven sub-fractions (Fr. 1D-10-A–1D-10-G) and compound 7 (42.0 mg). Fraction 1D-10-F was subjected to silica gel (1.0 cm × 80 cm) column chromatography with CHCl3–acetone (2.3:1; 1 L) as elution solvent to give compounds 6 (7.4 mg) and 8 (19.0 mg). Fraction 1E was separated using YMC (1.0 cm × 80 cm) column chromatography with MeOH–H2O mixtures (1:1, 3:1, 5:1, 7.5:1, 9:1, 15:1; 300 mL for each step) as eluent to give compound 2 (11.8 mg).
3.4. Product Characterization
Cerevisterol 6-cinnamate (6): Yellowish oil; C12H12O5; [α]: −21.8 (c 0.2, MeOH); 1H-NMR (CDCl3, 600 MHz) and 13C-NMR data (CDCl3, 150 MHz), see Table 1; HR-ESI-MS (C37H52O4): m/z 561.3944 [M + H]+ (calcd. 561.3938).
3.5. Cell Toxicity Assay
Cell-Counting Kit (CCK)-8 (Dojindo, Kumamoto, Japan) was used to analyze the effect of compounds on cell toxicity according to the manufacturer's instructions. Cells were cultured overnight in a 96-well plate (~1 × 104 cells/well). Cell toxicity was assessed after the addition of compounds on dose-dependent manner. After 24 h of treatment, 10 μL of the CCK-8 solution was added to triplicate wells, and incubated for 1 h. Absorbance was measured at 450 nm to determine viable cell numbers in wells.
3.6. Oxygen Radical Absorbance Capacity (ORAC) Assay
The assay was performed on a Tecan GENios multi-functional plate reader (Tecan, Salzburg, Austria) with fluorescent filters using excitation and emission wavelengths of 485 and 535 nm, respectively. In the final assay mixture, 40 nM fluorescein was used as a target of free radical attack with 20 mM 2,2-azobis dihydrochloride (AAPH) as a peroxyl radical generator [26]. Trolox (1.0 μM) was used as a control standard and prepared fresh daily. The analyzer was programmed to record the fluorescence of fluorescein every 2 min after the addition of AAPH. Fluorescein (40 nM) in potassium phosphate buffer was used as the blank. All fluorescence measurements were expressed relative to the initial reading. Final results were calculated based on the difference in area under the fluorescence decay curve between the blank and each sample. ORACROO· was expressed as micromoles of Trolox equivalents (TE). One ORAC unit is equivalent to the net protection area of 1.0 μM of Trolox.
3.7. Reduction Capacity
The reducing abilities of isolated compounds were determined according to the CUPRAC assay [27]. A total of 40 μL of different concentrations of the compounds in distilled water were mixed with 160 μL of a mixture containing 0.5 mM CuCl2 and 0.75 mM neocuproine in 10 mM phosphate buffer, pH 7.4. The absorbance was measured using a micro-plate reader at 454 nm for 1 h. The increase in the absorbance of the reaction mixture was indicative of increased reducing power. Distilled water was used as the blank.
3.8. Cellular Antioxidant Capacity
Cellular oxidative stress due to ROS generated by AAPH was measured spectrofluorometrically using the DCFH-DA method [28]. DCFH-DA diffuses through the cell membrane and is enzymatically hydrolyzed by intracellular esterase to non-fluorescent DCFH, which is rapidly oxidized to highly fluorescent DCF in the presence of ROS. HepG2 cells were first cultured in 96-well plates (5 × 105/mL) with DMEM for 24 h. After the cells were incubated with different concentrations of extracts and isolated compounds dissolved in DMSO for 30 min, the medium was discarded, and the wells were gently washed twice with PBS. HBSS, which is fluorescently stable, was then added to each well instead of normal medium, and AAPH was used as an oxidative stress inducer. After the cells were treated with 60 μM AAPH for 30 min, DCFHDA was added to the culture plates at a final concentration of 40 μM and incubated for 30 min at 37 °C in the dark. Trolox (10 μM) was used as the positive control. After the incubation, the cells were washed with HBSS, and DCF fluorescence intensity was measured at an excitation wavelength of 485 nm and an emission wavelength of 535 nm using a Tecan GENios fluorometric plate reader. DMEM medium was used as the control.
3.9. Tartrate-resistant Acid Phosphatase (TRAP) Activity
RAW264.7 cells were seeded in 96-well plates (1 × 104 cells/well) containing DMEM medium plus 10% fetal bovine serum (FBS) and the medium was replaced with test samples in a differentiation medium containing 50 ng/mL RANKL. After differentiating the RAW 264.7 cells into osteoclasts for 5 days, the medium was removed, and the cell monolayer was gently washed twice using ice-cold PBS. The cells were fixed in 3.5% formaldehyde for 10 min and ethanol–acetone (1:1) for 1 min. Subsequently, the dried cells were incubated in 50 mM citrate buffer (pH 4.5) containing 10 mM sodium tartrate and 6 mM p-nitrophenylphosphate (PNPP). After 1 h incubation, the reaction mixtures were transferred to new well plates containing an equal volume of 0.1 N NaOH. Absorbance was measured at 408 nm using an ELISA reader. Control was treated with only DMEM medium and treated control was treated with DMEM medium containing 50 ng/mL RANKL. TRAP activity was expressed as a percentage of the control.
4. Conclusions
In this study, a new sterol (cerevisterol 6-cinnamate; 6), together with 10 known compounds 1–5 and 7–11 were isolated from the fruiting bodies of H. erinaceum. All compounds exhibited peroxyl radical-scavenging capacity, while compounds 1, 3, and 4 showed reducing capacity too. Moreover, some sterols showed moderate effects on cellular antioxidant activity and inhibited RANKL-induced osteoclastic differentiation. To the best of our knowledge, this is the first report on the antioxidant and anti-osteoporotic activities of aromatic compounds and sterol components from H. erinaceum. These results provide a scientific basis for the development of novel antioxidant and anti-osteoporotic agents from this mushroom.
Supplementary Materials
The following are available online at: https://www.mdpi.com/1420-3049/22/1/108/s1, The NMR (1D and 2D) data of compound 6 are available as supporting information.
This research was supported by Chungnam National University and a grant (K17281) awarded to the Korean Institute of Oriental Medicine by the Ministry of Education, Science and Technology (MEST), Korea.
Author Contributions
W. Li performed the isolation, structure elucidation of the constituents, and prepared the manuscript. J.Y. Ma contributed to the revision of this manuscript. S.H. Lee and H.D. Jang conducted the bioassay experiments. The whole research was performed based on the planning of Y.H. Kim. All authors approved the final version of the manuscript.
Conflicts of Interest
The authors declare no competing financial interest.
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- Sample Availability: Samples of the compounds are available from the authors.
Figure 3. Peroxyl radical-scavenging capacity ((A,B)) and reducing capacity ((C,D)) of the extracts and compounds 1–11 from H. erinaceum. Data are expressed as the mean ± standard deviation of three individual experiments. Statistical significance is determined by one-way ANOVA followed by Dunnett's multiple comparison test (p < 0.05).
Figure 4. Cellular antioxidant capacity of compounds 1–11 against oxidative stress induced by 2,2'-azobisisobutyramidinium chloride (APPH) (Con.: control). Data are expressed as percentages of the value of untreated cells (mean ± standard deviation of three individual experiments). Statistical significance is determined by one-way ANOVA followed by Dunnett's multiple comparison test (p < 0.05).
Figure 5. Inhibitory effects of the extracts (A) and compounds 1–11 (B) on TRAP activity of osteoclastic RAW 264.7 cells. Data are expressed as percentages of the values obtained with untreated cells. (Con.: control, which was not treated; TC: treated control, which was treated with RANKL).
Table 1. 1H (600 MHz) and 13C-NMR (150 MHz) spectroscopic data of compound 6 (CDCl3, δ (ppm), J (Hz)).
|1.57, m
|1.99, m
|1.97, m
|1.41, m
|0.95, d, 6.5
|4.02, m
|5.09, dd, 15.3, 7.7
|1.69, m
|5.14, dd, 15.3, 7.0
|1.87, m
|1.80, m
|4.93, d, 5.4
|1.40, m
|5.26, d, 5.4
|0.76, d, 6.5
|0.76, d, 6.5
|1.94, m
|0.84, d, 6.7
|1.55, m
|6.33, d, 16.0
|1.27, m
|7.58, d, 16.0
|1.68, m
|1.87, m
|7.46, d, 8.0
|1.35, m
|7.30–7.31, m
|1.51, m
|1.21, m
|7.30–7.31, m
|1.67, m
|1.22, m
|7.30–7.31, m
|0.53, s
|7.46, d, 8.0
|1.08, s
© 2017 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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De Bruid: In a very special way the bride is being introduced to her father in law. Colorfully illustrated by Nanne Meulendijks with Dutch text.
Auhtor: Herman Pieter de Boer (1928-2014) was a Dutch writer, journalist and songwriter. He wrote poems, lyrics for radio and television and cabaret programs. Famous songs from his hand are 'Visite' (sung by Lenny Kuhr), 'Op een onbewoond eiland' and 'Ik heb zo waanzinnig gedroomd', wrtitten for 'Kinderen voor Kinderen'. De Boer also wrote the text 'Oh Waterlooplein' (1969), under the pseudonym Johnny Austerlitz.
Illustrator: Nanne Meulendijks is an independent artist and quickly reached the top in Holland as an illustrator. Her work appears in nationwide newspapers such as NRC Handelsblad, NRC Next, AD, Trouw and Volkskrant. She is working as well for the publishing firm Lannoo (Children's books).


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The Public Utilities Board (PUB) review aimed at auto insurance reform in the province is long over, yet the board is still receiving correspondence from the two main adversaries that fought it out during the review.
In fact, both the Insurance Bureau of Canada (IBC) — the association that represents insurers— and a group of personal injury lawyers in the province that formed the Insult To Injury Campaign aimed at protecting accident victims, have written the PUB in recent weeks slamming each other.
The PUB's review had looked into the reasons behind increasing claims costs for private passenger vehicles and taxi operators, and looked at options to reduce these costs. The board had been specifically asked to examine the impact on rates and implications for claimants of introducing a monetary cap on claims for non-economic loss for minor injuries or continuing with the current deductible of $2,500 or increasing the deductible.
The PUB presented its report to the provincial government on Jan. 29.
In the House of Assembly in its current sitting, Service NL Minister Sherry Gambin-Walsh stated her department has already implemented a number of measures to help strengthen the taxi industry in relation to auto insurance.
Measures for other drivers are to follow in the coming weeks.
"Effective July 1, 2018, there were policy amendments focusing on skilled drivers, experience in driving in provincial road conditions, driving history, and passenger safety and safer vehicles," the minister said. "A zero tolerance policy for drugs or drugs, and alcohol when operating a taxi, came into effect on Dec. 18, 2018.
"In Budget 2018, government approved a one-third reduction in the tax on auto insurance over four years. This will particularly benefit those paying the highest rates, such as taxi operators. Our government remains committed to working with stakeholders, and bringing forward changes to legislation during this sitting of the House that will benefit consumers of insurance in Newfoundland and Labrador."
During the PUB hearings, the IBC has stated that the imposition of a $5,000 minor injury cap, plus other reforms that will give accident victims faster access to health care and a quicker recovery, is the best option to stabilize insurance premiums and one day see a reduction in rates in Newfoundland and Labrador.
The Insult To Injury Campaign, however, argued that if a minor injury cap is imposed, accident victims will lose the right to sue for fair compensation and premiums will not go down and likely continue to rise.
The letters to the PUB came after a CBC News report stated that insurance premiums in New Brunswick have been dramatically increasing the last couple of years despite a minor injury cap in place in that province.
The Insult To Injury Campaign wrote the PUB suggesting that the IBC, in its filings with the board and throughout the hearings, continued to provide and rely on 2016 and earlier data from other Atlantic Provinces in support of its argument that a cap on minor injuries in New Brunswick and Nova Scotia was an example of a mechanism for controlling claims costs and providing rate stability.
"All the while, the IBC was well aware that rates in New Brunswick had begun to escalate to a significant degree after 2016 and did nothing to bring this to the attention of the board," the lawyers' letter stated.
The lawyers asked the PUB to denounce in writing "in the strongest possible terms" the conduct of the IBC uncovered in the news report.
Amanda Dean, IBC's vice-president Atlantic, however, fired off a letter to the PUB in response. She said the rate filings in New Brunswick are posted to the New Brunswick Insurance Board website and publicly available.
"In reference to the use of 2016 data during the hearing instead of 2017, IBC only obtained the 2017 GISA (General Insurance Statistical Agency) data upon its public release by GISA in July of 2018," Dean wrote.
"The 2017 numbers referenced in the PUB report were arrived at through the actuarial work of Oliver Wyman, and included additional GISA information to which IBC did not have access.
"It is of note to recognize that in 2017 the average written premium in New Brunswick was $819, while the average written premium in Newfoundland and Labrador was $1,132."
She said IBC takes great issue with the assertion it deliberately misled the PUB hearings.
The province's consumer advocate, Dennis Browne, has stated it is difficult to conclude that the introduction of a cap would be an improvement or translate into long-term, stable rates for consumers.
He prefers the option of increasing the current deductible of $2,500 to $10,000.
In a recent letter to The Telegram, Browne stated that:
"The purpose of the current $2,500 deductible was to stabilize and reduce rates, but that purpose has not been fulfilled. A deductible regime in the $10,000 range may be one method to ensure reduced, stable and affordable premiums. In introducing the new regime, a period of 24 months may be appropriate for a rate freeze after which time an assessment can be made to see if the $10,000.00 deductible is having the intended effect."