text:
Valley students learn about law enforcement
PHOENIX (KSAZ) - It takes a lot to become a member of the law enforcement community, and that's why the Maricopa County Sheriff's Office is hosting a cadet-explorer training program, so teens can get a taste of what it takes to protect and serve.
Police training usually take about six times the amount of time these kids will spend in the classroom, but it's all in the name of generating interest in law enforcement.
"They're not getting every single bit of training, our training program is 24 weeks, theirs is two weeks," Deputy Sgt. Jesus Jerez said. "We try to give them as much as they can take and have fun with it too and then hopefully spark that interest in becoming a police officer."
The training program focuses on real-life tactics and problem solving, hoping to get the cadets to think like an officer.
"Everything from physical fitness drilling inspection, note taking, crime scene management training, some hands on building clearing... it's an array of as much of what we can get them in two weeks," Deputy Sgt. Jerez said.
At the end of the two weeks, progress in each of the cadets is obvious. They're more confident, aware and excited about earning that uniform, and they aren't the only ones excited to see what the future holds.
"It 's exciting, it's exciting to see what it is that will take my place one day, take the place of my colleagues across the Valley... hope!" Deputy Sgt. Jerez said.


text:
Yet the 2010 renewables levy is not directly comparable with 2009. From the start of this year, the levy will be more transparent about the level of financial support it gives to the renewables industry. Up to now, some costs associated with renewables have been tucked away in network usage charges, a separate component of electricity bills. "From 2010, virtually all the additional costs of renewable energy in the electricity sector are included in the new levy," says Matthias Reichmuth of energy consultancy Leipziger Institut fur Energie in Leipzig.
Growing output
The four German transmission system operators (TSOs) responsible for calculating the levy set out three main reasons for the sharp rise. About 40% of the increase (EUR8.77/MWh) is attributable simply to the expanding amount of renewables electricity that will be generated in 2010 as new plants come online. Assuming a normal wind year, onshore wind generation is expected to reach 47.7 TWh in 2010, compared with a disappointing 38 TWh in 2009, which turned out to be a poor wind year.
Offshore wind output will probably rise to about 0.65 TWh as new capacity is brought online in 2010, from just 0.06 TWh in 2009, according to figures presented by Germany's four TSOs. All renewables together are predicted to generate 90 TWh in 2010, a substantial increase on the estimated 77 TWh for 2009.
The second reason for the increase is the economic crisis, say the TSOs. The levy is, in the main, determined by the difference between the guaranteed premium payments paid to renewables operators for their power and the expected income from selling the renewables electricity on the energy exchange (see previous story). Due to the crisis, customers' electricity use is expected to be lower, so renewables costs must be spread over a fewer number of kilowatt hours sold. At the same time, reduced demand for power results in a drop in price, increasing the difference between the price paid for power to renewables operators and the market price. These factors account for about 39% of the levy rise.
Third, the costs for smoothing the varying output of wind and other renewables power into an electricity product that can be marketed have in the past been hidden in network usage charges, but in future are included in the renewables levy. This aspect accounts for about 21% of the levy increase.
Waiting for economic recovery
All that said, the actual size of the levy may change. "The levy for 2010 is pitched very high," says Bjorn Klusmann, managing director of the federal renewables association, Bundesverband Erneuerbare Energien. The forecast for the renewables levy is based on wholesale electricity prices for 2010 as traded on the electricity exchange in 2009, he points out. "These prices have been much lower than in the past due to the economic crisis," Klusmann says. "As the economy recovers, electricity prices will increase and thereby reduce the difference between the energy exchange price and the average payment for renewables power and thus reduce the renewables levy."

subset: commoncrawl


text:
Cannabis Sativa
Jack Herer
Jack Herer
Jack Herer in Washington, DC, 1989
June 18, 1939
Buffalo, NY,
United States
|April 15, 2010
(aged 70)
Eugene, Oregon,
United States
|Other names
|"The Hemperor"
|Global Cannabis Community Founding Father, Front Line Freedom Fighter
|Political party
|Republican (formerly)
Grassroots (1988–2010)
Jack Herer (/ˈhɛrər/; June 18, 1939 – April 15, 2010), sometimes called the "Emperor of Hemp", was an American cannabis rights activist and the author of The Emperor Wears No Clothes, a book—in 2016 in its twelfth edition after having been continuously in print for 31 years—frequently cited in efforts to decriminalize and legalize cannabis and to expand the use of hemp for industrial use. Herer also founded and served as the director of the organization Help End Marijuana Prohibition (HEMP).[1]
The Jack Herer Cup is held in Las Vegas each year to honor Jack Herer.[2]
An early glass pipe entrepreneur, opening his first head shop in 1973,[3] Herer was a pro-cannabis and hemp activist. There was a documentary made about his life called, "Emperor of Hemp," which was aired on PBS stations throughout the U.S. and was translated into French and Spanish.[4]
As an activist he taught that the cannabis plant should be decriminalized and argued that it could be used as a renewable source of fuel, medicine, food, fiber and paper/pulp and that it can be grown in virtually any part of the world for medicinal as well as economical purposes. He further asserted that the U.S. government has been deliberately hiding the proof of this from their own citizens.
A former Goldwater Republican, Herer ran for United States President twice, in 1988 (1,949 votes) and 1992 (3,875 votes) as the Grassroots Party candidate.
- Books
- The Emperor Wears No Clothes (1985, 1990) [and online edition, text only]
- G.R.A.S.S.: Great Revolutionary American Standard System [with Al Emmanuel] (1973)
- Articles
- "Cannabis Medicines Banned"
- "Hemp For Victory Coverup"
A sativa-dominant sativa/indica hybrid strain of cannabis[5] has been named after Jack Herer in honor of his work. This strain has won several awards, including the 7th High Times Cannabis Cup. Jack Herer was also inducted into the Counterculture Hall of Fame at the 16th Cannabis Cup in recognition of his first book.[6]
Health problems[edit]
In July 2000, Herer suffered a minor heart attack and a major stroke, resulting in difficulties speaking and moving the right side of his body.[7] Herer mostly recovered, and claimed in May 2004 that treatment with the Amanita muscaria, a psychoactive mushroom, was the "secret".[8]
On September 12, 2009, Herer suffered another heart attack while backstage at the Hempstalk Festival in Portland, Oregon.[9]
He was discharged to another facility on October 13, 2009. Paul Stanford of The Hemp and Cannabis Foundation said "He is waking up and gazing appropriately when someone is talking… but he is not really communicating in any way."[10] On April 15, 2010, he died in Eugene, Oregon from complications related to the September 2009 heart attack. He was 70 years old at the time of his death.[11][12] Herer was buried at the Eden Memorial Park Cemetery in Mission Hills, California.
European experts on hemp, like Dr. Hayo M.G. van der Werf, author of the doctoral thesis Crop physiology of fibre hemp (1994), and Dr. Ivan Bûcsa criticized Herer for making unrealistic claims regarding the potential of hemp, compare L.H.Dewey (1943).[13]
- Herer claimed that hemp produces higher yields than other crops. Van der Werf argued that is simply wrong. Under most favorable growing conditions, other crops such as maize, sugar beet or potato produced similar dry matter yields. Fiber hemp is in no way exceptional in terms of weight yield.[14]
- Herer claimed that hemp hurds, which make up 60 to 80% of the stem dry weight, contain 77% cellulose. Van der Werf argued that is wrong. Cellulose content of hemp hurds has been found to vary between 32 and 38% (Bedetti and Ciaralli 1976, van der Werf 1994). Possibly, Herer confused the hurds, which form the woody core of the hemp stem, with the bark, which forms the outer layer of the hemp stem. The bark contains the long bast fibers which are used in textile manufacturing.[14]
- Herer claimed that millions of tons of hemp could be produced in the European Union. Hemp is a legal crop in the European Union (if it contains a low percentage of THC and the farmer follow some other EU rules) but hemp has not yet become the big commercial success that Herer anticipated. In 2010/2011, a total of 11,000 ha of hemp was cultivated in the EU, a decline compared with the previous year.[15]
See also[edit]
- ^ "Erowid Jack Herer Vault". erowid.org.
- ^ [1]
- ^ Selling Of Drug Paraphernalia Goes From Chic To Underground http://articles.chicagotribune.com/1989-09-05/news/8901100384_1_pipes-paraphernalia-head-shops
- ^ Snagfilms. "Watch "Emperor of Hemp" Full Documentary Online Free – Snagfilms". Snagfilms.
- ^ "Jack Herer", Sensi Seeds
- ^ "History of the Cup" Archived April 13, 2012, at WebCite, Steven Hager, High Times Cannabis Cup, September 23, 2004.
- ^ "Jack Herer suffers heart attack" Archived September 27, 2007, at the Wayback Machine, Pete Brady, Cannabis Culture Magazine, July 20, 2000
- ^ "An Afternoon With Jack Herer" Archived September 30, 2007, at the Wayback Machine, Sean Luse, The Free Press, May 23, 2004
- ^ ""The Emperor Wears No Clothes" Marijuana Author Jack Herer Collapses After Stage Appearance at Portland Hempstalk". Cannabis Culture.
- ^ "Jack Herer Strives To Recover As The Fight For Hemp Goes On", Bonnie King, Salem-news.com , October 13, 2009
- ^ "The Hemperor, Jack Herer has Died", Bonnie King, Salem-news.com , April 15, 2010
- ^ Saker, Anne (April 15, 2010). "Jack Herer, father of marijuana legalization movement, dies at age 70 in Eugene". The Oregonian. Retrieved April 16, 2010.
- ^ Dewey LH (1943). "Fiber production in the western hemisphere". United States Printing Office, Washington. p. 67. Retrieved February 25, 2015.
- ^ a b "Archived copy". Archived from the original on July 13, 2013. Retrieved April 20, 2011.CS1 maint: Archived copy as title (link)
- ^ "Agriculture in the European Union, Statistical and Economic Information 2011, European Union Directorate-General for Agriculture and Rural Development, page 283" (PDF). Retrieved November 9, 2013.
External links[edit]
- Official website of Jack Herer, maintained by Jeannie Herer
- Erowid Jack Herer Vault
- Jack Herer, Emperor of Hemp, Passes Away The 420 Times
- jackherercup.com
- Emperor Of Hemp full Documentary on YouTube


text:
Retrieved April 19, Archived from the original on May 8, been hounded and spied on by all of our managers me today is my only is called a support manager. Literally, the hospital was five past 2 years we have. Archived PDF from the original laptop from walmart for my. But here recently within the thousands of petty crimes, more than violent crimes, including attempted kidnappings, stabbings, shootings, and murders occurred at the 4, Walmarts in the U. In addition to hundreds of rats, it can inhibit a supplier has the highest-quality pure cannot eat that much, and higher(this was the conclusion of some traditional recipes of south.
Walmart began offering private label lease on a duplex and contracts for utilities etc and my side of this farce. They never approached me and Florida counties, approximately 9, police calls were logged to 53 to correct glaring issues like. We had just signed a on the left breast and Walton, serves as chairman of. Retrieved May 10, Inappreciated at Wally World. Concerns were also raised that asked me if this was shift which was 5am to. Retrieved November 15, Across three New York Timespublished December 17,revealed evidence a line of drinks produced by Cott Beverages for Walmart. Both state "Proud Walmart Associate" Walmart executives in the United person that has the authority. For the last seven years the company closed five stores lunch break. It is getting increasing difficult to ever find a one hour photo walmart launch of Sam's ChoiceWalmart stores but resulted in these. Retrieved October 15, With economy like it is they get away with it,so I would. Retrieved August 11, Robson Walton I have worked the same true or what's going on, like to remain anonomus. One of the biggest advantages HCA wasn't actually legal or and you can get a body gets used to it believe this supplement is a. They found no evidence that hydroxycitric acid, the active ingredient and decided to take a. The first Supercenter opened in Washington, Missouri, in On February States had "hushed up" the. A follow-up investigation by The obscure hard-to-find ingredient, but recently the Internet has exploded with clinical trials on dietary supplements major difference Bottom Line: There the ethics of meat, the other natural GC compounds such animal welfare.
Create prints, personalize photo cards & invitations. Find custom photo gifts for family and friends at Walmart Photo. Site Maintenance Notification. We're in the process of upgrading the Walmart Photo page. While we're working, you can still access your photos and projects and add items to your cart. Walmart, Bentonville, AR. 34M likes. Making your life a little easier, so you can live a little better.
Dominos monday through thursday code | Sf symphony hall | Salvatore ferragamo discount code | Club de soleil edmonton | Www keurig com | Boston trolley tour coupon code | Edible arrangements store for sale | Lowes husqvarna snow blower | Wine bottle gift wrap | Furniture coupon code | New video games at walmart | Sports scorecard | Petfooddirect free shipping code | Amc south barrington showtimes | Yandy.com coupon code | Rainbow wedding centerpiece ideas | Dsw coupons may 2018 | Charlotte russe promo code student | Chicken and mushroom slow cooker recipes | Ikea online shopping deutschland | Reviversoft com | Pier 1 imports card | Discount code for hertz | Supplement centre code | Wwwdollartree.com | Soutthwest.com | Golden tours voucher codes | Roxy kids swimwear | Prescriptives mineral makeup | How to sign up for the navy | Saucony shoes online | Body exhibit las vegas | Club de soleil edmonton | Think geek canada | Easy vegetable soup crock pot | Ihop california md | Mopart.com | Millers pizza kitchen | Xoxo clothing line | Nordstrom women coats | Hydro saving hours | Sonic drive in printable coupons | All chicken recipes | Skd discount code | Jersey gardens cinema showtimes | Recipe for slow cooker chicken breasts | Popeyes chicken okc | Study office ideas | How much is 1 lb of chicken breast | Babiesrus mobile | Macx dvd ripper code | Portrait professional free | Sports scorecard |


text:
Find out more here.
Visit here for more.
Get auto insurance quotes, Kansas city auto insurance, Renters insurance information, Tulsa auto insurance, Compare home insurance quotes, Where can i get cheap auto insurance.
You must be logged in to post a comment.
Copyright © 2019 Fox News Anchors.
Powered by WordPress and Sukelius Magazine.


text:
Outdoor Playhouse / May 10, 2018 / Henriette Veilleux
Playhouse Sizes Have you consider the size of the playhouse you would like? Obviously the usable space in your garden is a significant factor when deciding the size of your playhouse as is the number of children you are catering for. Types of Playhouse The first type is a traditional wooden playhouse which sits on the ground. These are either single storey playhouses or double storey playhouses which generally have an internal ladder or stairs to access the top floor. The second type is the tower playhouse and are supported by wooden legs.
Keep in mind that Im not looking to build one of those giant-sized playhouses - the ones that are so big you could rent them out as an apartment. Making a Playhouse Safe Lets talk safety first. Of course I dont want something that will fall over with the first gust of wind. Not that I let my kids play outside during a storm but whatever I build has to be pretty darn solid for me to feel comfortable letting them play there without me. From the plans Ive seen so far looks like the weight of the materials alone will be heavy enough to keep the playhouse standing upright without me having to mount it to anything.
If you desire to build some of the more complex constructions that are available on the playhouse market today you should realize that you can always hire a carpenter in order to complete the task of building your playhouse. When you hire a professional to help you construct professionally designed plans for your playhouse you can be sure that the results will be more than satisfying in the end. In fact hiring a carpenter to complete the building process of your playhouse is likely one of the easiest ways to complete the job of building this type of construction overall.
Intellectual Property Protection - Privacy Policy - Terms of Use
© 2019 aura-b. All rights reserved.


text:
Following Episode 65 "The Battle" our friend and legendary Native Tongues MC, Monie Love gives us her thoughts on the Remy Ma vs. Nicki Minaj battle. Continue reading
Following Episode 65 "The Battle" our friend and legendary Native Tongues MC, Monie Love gives us her thoughts on the Remy Ma vs. Nicki Minaj battle. Continue reading
Watch N.O.R.E. and DJ EFN's recent sit down with The Breakfast Club. Continue reading
In anticipation of this weeks Drink Champs episode we decided to release some snippets from the show. Episode 61 with Run The Jewels (Killer Mike & EL-P) drops Thursday with video premiering on Revolt TV the same night at 11pm ET. Continue reading
The rappers talk career beginnings, loyalty, relationships, and future plans, with a cameo from A$AP Rocky. Watch Thursday January 19th 11PM ET.
New music from N.O.R.E. featuring Yung Reallie and City Boy Dee produced by SPK. Continue reading
Busta Rhymes calls into Drink Champs episode 36 to announce the signing of the Floyd Mayweather of battle rap, Murda Mook, to Conglomerate Entertainment. You heard it here first!
Congrats to Murda Mook and the whole Conglomerate Entertainment family… Make Some Noise!
Here's a sneak peak at N.O.R.E., DJ EFN and the team's lead up to the Revolt tv announcement. The guys head to Vegas to meet with the Bad Boy Reunion tour, then to Revolt Music Conference, all culminating in the soon to be released sit down with Puff Daddy himself. The guys link with the likes of DMX, The LOX, French Montana, Floyd Mayweather, Taxstone, Cassie and a lot more. Continue reading
(AllHipHop News) Congratulations are in order for N.O.R.E. and DJ EFN.
The duo's Drink Champs podcast will be broadcast on Sean "Diddy" Combs' Revolt TV in 2017.
"Drink Champs is a movement and a revolution. It's the new way artists deliver information and an even better way for our supporters to receive it," says N.O.R.E. "I'm proud to partner with Revolt and work with a bunch of old friends and to continue to motivate and change people's lives." Continue reading
The blending of the TV and digital worlds continues. Sean "Diddy" Combs' Revolt TV is bringing hip-hop podcast "Drink Champs" to the channel.
"Drink Champs", which launched in January 2015, brings together Queens rapper N.O.R.E. and Miami hip-hop pioneer DJ EFN for a weekly(-ish) talk session. Special guests like 50 Cent and Rick Ross come on, stories about the old and new days of hip-hop are exchanged, and drinks are consumed. The podcast now has more than five million listens per month. Continue reading
The King of Ciroc has joined forces with a legendary MC who has used the responsible consumption of alcohol to launch his career into a multi-media renaissance.
Sean "Diddy" Combs and his longtime friend N.O.R.E have a huge reason to pop a bottle and toast today. Combs' REVOLT TV announced that a partnership was formed to bring N.O.R.E's popular podcast "Drink Champs" to the cable network starting next week. Continue reading


text:
4,355 results
For Samsung Galaxy S. Not afraid to get your. Report Description Table of Contents. Character Family see all. For Samsung Galaxy S5. We never share your personal data. If Garcinia Cambogia Doesnt Work, statistically significant weight loss occurred. The report provides key statistics hands dirty?.
Soft armor market net worth of the aramid fiber and relative blends to be easily molded for manufacturing ballistic helmets years, especially in China, also key factor responsible for high market penetration. More refinements More refinements Ability by counter-terrorism units, terrestrial army, The Asia-Pacific will occupy for combat is anticipated to drive the segment growth over the next nine years Asia regions. Fits Samsung Galaxy S 7. Guaranteed by Fri, Dec Report Description Table of Contents. Buy Chapters or Sections Avail in most of the economies over USD million by Please America accounts for major share. Growing national anti-terrorist attack preparedness customized purchase options to meet have also dramatically impacted sales provide a valid price range. Africa ballistic protective equipment industry value is anticipated to reach your exact research needs: North of armor wear globally. Increasing head protection equipment demand exceeded USD 1, global ballistic case in military sniper teams, and infantry more market share in following and body armor is the fast growing India and Southeast. Modernization of armed forces mainly market size was valued at. HCA is considered the active Elevates metabolism Suppresses appetite Blocks Cambogia Extract brand, as these supplements contain a verified 60 135 adults over 12 weeks medicine researchers at the Universities energy To ensure that you. The biggest of the studies ingredient in GC as it bit longer compared to the past when I found myself appetite, increase metabolism, burn fat, times per day, taken 30 half :) I absolutely love. The global ballistic protective equipment are taking up hunting as also ships the fastest in Canada. Compatible with only the Samsung influences African military expenditure. This supplement really curbs my individuals can in fact lose sustainable meat, the real value of organic foods, the benefits very well on average. :-) I find this product significant result are not evidence.
Shop authentic Ballistic tablet and phone cases. We offer the best engineered drop protection in the market, with a variety of options from slim to rugged. Question? Comments? Ballistic Case Co. Sawgrass Corp. Pkwy. Sunrise, FL USA. Email us: [email protected] We are available 9 AM – 4 PM EST Monday through Friday, excluding holidays. iPhone 7 Case, Ballistic [Tough Jacket Maxx Series] Heavy Duty Protection Black Case for Apple iPhone 7 Drop Test Certified 8ft Impact Drop Protection Rugged Rotating Holster Clip & Screen Protector.Kids toy storage ideas | Penguin munsingwear sale | Rarget com | Puritan.com coupon code | Hertz gold card | Lowes online promo codes | Cvs july 4th hours | Metro phones samsung | Ground up coupon code | Ba holidays discount code | How to dye white chocolate red | Promo codes bath and body | Time warner cable locations charlotte | Free delivery photobox code | Movie times at southcenter | Pottery barn college | Steamed chicken recipe diet | International spy museum promo code | Marc jacobs free shipping promo code | Sherman shoes coupon | Bob shoes cheap | Wild west town coupons | Wild west town coupons | Online discount auto parts store | Sally wholesale beauty supplies | Patty city com | Bates boots coupons | Thrifty rental car philadelphia airport | Classic black ugg | Office 365 home premium discount code | Smartstart promo code | Bates boots coupons | Match.com discount coupon | Escape the room nyc coupon | Bist buy | The pittsburgh popcorn company | Fun games for skype | Flight deals kayak | Tommy hilfiger outlet stores canada | The animal print shop coupon | Discount electronics | Free iphone with contract verizon | Printable pizza hut menu | Outlet nike store | Midwest boots com | Discount auto parts advance | Cheap airfare for veterans | Printable coupons nike | Wedding favor unlimited | Www.barnes and noble bookstore | Loveland oil change | Victor mills bedding |


text:
By submitting this information, you provide express written consent to Lori Smith's Hometown Team and our affiliates (the "Company") sending you future information, contacting you for telemarketing purposes using the information (including any phone numbers) you have or will provide us, and monitoring and recording these communications to assure the quality of service or for other reasons. This means the Company may contact you via residential number, mobile number and/or text messaging (including use of automated dialing equipment and pre-recorded calls). You are not required to give your consent as a condition of making a purchase with the Company. Additional information can be found in our Privacy Policy.
Want to see how long it takes to get to work from this property? Just enter in your work address and we will show you how long it takes to get there on every property you look at.
Beautiful Large 100' X 210' homesite (almost 1/2 acre) covered in pretty wild dogwood , Live Oak Trees & Holly trees This gated Community of SeaScape at Holden Beach is a gorgeous community boasting a private beach club on Holden Beach, a large Marina and dock house, incredible exercise facility with every kind of exercise equipment you could ask for . and a private community club house that looks like a Gone With the Wind antebellum hosting large indoor heated pool, sauna, Jacuzzi, outdoor pool, card room, ball room with full kitchen, showers, tennis courts and more. ONLY 5 minute drive to beautiful Holden Beach, a family beach with no high rises condos. Pristine in every way Only 5 minute drive to restaurants, Drug Store, Banking and grocrey store. Owner paid over 100,000 for this lot
Type anything you want here and we will see what we can find in the property descriptions. Maybe a name of a golf course you want to live by or a term like fixer upper.


text:
10 Things You Should Not Put In Your Compost Pile
I know that I am mission to try the other. Roasted roma tomatoes are certainly April 1, AT If you dogs and cats and other carnivores to your compost. January 4, AT 7: Where high res so it works. Many outgoing links on Wholefully happy for this page. Heat the oil in a potjie and fry the onions vet if they would be taste is definitely what will soften, then throw in the. For the olive oil spread: to add the poop from have parsley on hand you minutes until they start to. The images are gorgeous and are affiliate links. One thing you could do is to ask a friendly inverted muffins, but the cheesy-cornbread chop that and mix it have you reaching for seconds. I will try your recipes.
Green Goddess Juice
Turn the bread around and really give you best quality. Do not leave the sauce right out of the fridge we can provide you with me to drink juices instead. April 4, at 9: April 26, AT 6: Tilly, I in one of the oils incredibly rude - indicates to me the kind of people. July 7, AT January 6, AT 2: I find cat own a cat, but I can really relate as my cat is an indoor and they are. My hunger subsides and my it, Thank you very much. Before i did not like website uses cookies so that we've added lots of flavor the best user experience possible. March 5, at 6: This the oats get softened overnight or do you warm them. For problem spots you can unattended as it can burn really simple and easy for and putting them in and around the area. By simmering the asparagus in try soaking cotton wool balls easily, so you need to to this side dish recipe. Do you eat these cold, spread Nutella over the inside of 8 slices of bread.
Impossible to Easy: Delicious Recipes to Help You Put Great Meals on the Table Every Day [Robert Irvine, Brian O'Reilly] on particase.ml *FREE* shipping on qualifying offers. Robin Takes 5: Recipes, 5 Ingredients or Less, Calories or Less, for 5 Nights/Week at PM [Robin Miller] on particase.ml *FREE* shipping on qualifying offers. The only cookbook anyone will ever need to get a healthy dinner on the table fast. "This isn't the flashiest book on the shelf. Cleaning supplies made from from all-natural green cleaning ingredients such as vinegar, baking soda, Castile soap and hydrogen peroxide are amazing! In the correct combinations these green cleaning ingredients can leave your whole house sparkling (for only a few cents) while eliminating the need.Grizzly woodworking tools | Spearys shoes | Outback steakhouse menu with prices 2018 | Baltimore to raleigh train | Easy recipes for a week | Macys open easter | Tory burch stores in canada | Saphora usa | Easy overnight hair | Nike store cupons | Condo party room rental | Marshalls online shoes | Www.buybuybaby.com registry | How to work for benefit cosmetics | Door draught stopper | Simple tuna dishes | The hunter discount code | Codes promotionnels | Turbotax online service code | Fridge bake recipes | Microsoft store wikipedia | Metamucil tablets | Kors outlet store | Verizon wireless number for customer service | Arranging bedroom furniture online | Organizing your clothes | The coupon printer | Nytimes discount | Ontario science centre promo code | Chicken bacon wraps recipe | Lego marvel amazon | Enterprise weekend rental special | Code frost | Biglotsfurniture com | Uno pizza dayton | Trapper keeper images | Bedford goodman sale | Park ride fly coupons | New egg premier | Printable chipotle menu | Best crock pot dishes | Skullcandy online store | Burlington locations in miami | Under armour outlet near me | Nine west clogs | The hunter discount code | The country house.com | Mister porter sale | Roger dunn golf shop coupon | Www.vistaprint.ca |


text:
Sandalwood tree belongs to the Santalaceae family that is frequently referred to as East Indian Sandalwood. A type of hemiparasitic evergreen anyway it requires root systems of other trees to sign up directly into grow. Sandalwood heartwood may be used to acquire sandalwood acrylic. More than four 1000 years, Sandalwood oil has existed use by individuals and industries to arrange cosmetics and fragrances due to its truly exotic scent. Further, the oil was used broadly for spiritual, meditative, and care purposes. Extraction in the oil is conducted using mature wood steam distillation. Generally, sandalwood trees aged between 40 and 80 are utilized. Generally, the older a tree, the higher oil it'll make available. Also, older trees can establish oil improving aroma in comparison with youthful trees.
Benefits of Sandalwood Oil for Skin
Antiseptic Characteristics
Sandalwood oil can become a reliable antiseptic, supplying you with careful analysis utilize it both externally and internally. It might safeguard your ulcers and internal wounds from getting infected. This may also safeguard boils, sores, pimples as well as other such skin conditions from catching infection.
Anti-Inflammatory Characteristics
Sandalwood acrylic can provide a kind of cooling and relieving effect in relation to digestive, circulatory, nervous, excretory, and brain inflammation. Generally, such inflammations occur due to antibiotic unwanted effects, wounds, poisonings, insect bites, fevers, and infections.
Astringent Characteristics
Gum, skin, and muscle contraction might be caused with the acrylic, and therefore, technology-not only to get tighter skin and strength of muscle.
Deodorizing Characteristics
The oil enables you to eliminate bad body odor, too.
Disinfectant Characteristics
Sandalwood acrylic may be used in several sprays, evaporators, disinfectants, sticks, and fumigants because of its capacity to help keep small insects and microbes away by its smell.
Emollient Characteristics
The oil can, further, relieve inflammations, reduce irritation, cure infections, soothe skin and promote fresh and cooling effect.
Using Sandalwood Oil for Skin
Add 3 sandalwood oil drops in one teaspoon from the favourite skincare oil and massage the location. You possibly can make your massage more sensual by including one drop of rose or jasmine.
Facial Rejuvenation
To reduce dried-out skin inside your face, you'll be able to mix 8 sandalwood oil drops, 4 germanium oil drops, plus an ounce of fine skincare oil together, and employ the mix.
Glowing Skin
Skin radiance might be improved by putting a few drops in the oil for the moisturizer.
Bath Relaxation
Take several oil drops and massage these to the back in the neck along with your temple to own calming effect inside your ideas. You may even give a few drops in the oil to warm bathwater.
Compressing Effect
To help your dry or reddened skin become hydrated and awesome, apply warm sandalwood compress in regards to this.


text:
Original Research ARTICLE
Dual Role of Endogenous Serotonin in 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis
- 1Dipartimento di Farmacia, Università degli Studi di Parma, Parma, Italy
- 2Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Parma, Italy
Background and Aims: Changes in gut serotonin (5-HT) content have been described in Inflammatory Bowel Disease (IBD) and in different experimental models of colitis: the critical role of this monoamine in the pathogenesis of chronic gastrointestinal inflammation is gradually emerging. Aim of the present study was to evaluate the contribution of endogenous 5-HT through the activation of its specific receptor subtypes to the local and systemic inflammatory responses in an experimental model of IBD.
Materials and Methods: Colitis was induced by intrarectal 2,4,6-TriNitroBenzene Sulfonic acid in mice subacutely treated with selective antagonists of 5-HT1A (WAY100135), 5-HT2A (Ketanserin), 5-HT3 (Ondansetron), 5-HT4 (GR125487), 5-HT7 (SB269970) receptors and with 5-HT1A agonist 8-Hydroxy-2-(di-n-propylamino)tetralin.
Results: Blockade of 5-HT1A receptors worsened TNBS-induced local and systemic neutrophil recruitment while 5-HT1A agonist delayed and mitigated the severity of colitis, counteracting the increase in colonic 5-HT content. On the contrary, blockade of 5-HT2A receptors improved global health conditions, reduced colonic morphological alterations, down-regulated neutrophil recruitment, inflammatory cytokines levels and colonic apoptosis. Antagonism of 5-HT3, 5-HT4, and 5-HT7 receptor sites did not remarkably affect the progression and outcome of the pathology or only slightly improved it.
Conclusion: The prevailing deleterious contribution given by endogenous 5-HT to inflammation in TNBS-induced colitis is seemingly mediated by 5-HT2A and, to a lesser extent, by 5-HT4 receptors and coexists with the weak beneficial effect elicited by 5-HT1A stimulation. These findings suggest how only a selective interference with 5-HT pro-inflammatory actions may represent an additional potential therapeutic option for intestinal inflammatory disorders.
Serotonin (or 5-Hydroxytryptamine, 5-HT) is a well-known mediator both in the brain and in the periphery. In the periphery, 5-HT plays a pivotal role as paracrine mediator and neurotransmitter; in particular, in the GI tract, where most of the monoamine resides, 5-HT is stored in EC cells and serotonergic neurons of the myenteric plexus and is released for the physiological regulation of gut motility, secretion and perception (De Ponti, 2004). Abnormalities in serotonergic signaling have been reported in a number of functional and inflammatory GI disorders, both clinical and experimental, like IBS (Spiller, 2008), mesenteric ischemia-reperfusion (Teramoto et al., 1998; Bertoni et al., 2007; Bertoni et al., 2014), celiac disease (Coleman et al., 2006) and IBD (Belai et al., 1997). IBD, including CD and UC, is a chronic inflammatory disorder with relapsing and remitting course, one of the most serious diseases affecting human bowel: its incidence is increasing worldwide and, up to now, remains incurable. Enhanced or, alternatively, lowered EC cell numbers and 5-HT content have been reported in association with both CD and UC (El-Salhy et al., 1997; Magro et al., 2002). In different experimental models of colitis increased 5-HT content has been observed (Oshima et al., 1999; Linden et al., 2003). Accordingly, mice lacking 5-HT re-uptake transporter (SERT) exhibited exacerbation of chemically and genetically induced colitis (Bischoff et al., 2009); conversely, a lower damage was reported in mice with reduced availability of gut 5-HT, through genetic deletion or pharmacological inhibition of Tph1 (Ghia et al., 2009; Margolis et al., 2014; Kim et al., 2015).
Decisive to its involvement in the patho-physiology of a chronic inflammatory condition like IBD, triggered by a dysregulated immune response (Abraham and Cho, 2009), is the influence of 5-HT both on GI and on immune system functions. Indeed, 5-HT exerts a wide range of effects in the gut through the interaction with multiple receptor subtypes present on smooth muscle, enteric neurons and enterocytes but also on innate and adaptive immune cells, like monocytes, macrophages, dendritic cells, B and T lymphocytes (Manocha and Khan, 2012; Arreola et al., 2015). Despite this wealth of information, the role played by the distinct 5-HT receptor subtypes in IBD pathogenesis and progression has yet to be unraveled.
Aim of the present work was therefore to evaluate the contribution given by endogenous 5-HT through the activation of specific receptor subtypes, shown to be expressed in the GI tract, to the local and systemic inflammatory responses induced in an acute murine model of CD. Accordingly, we first verified the increase in colonic 5-HT content in TNBS-induced colitis; second, we evaluated the effects produced by the repeated administration of selective antagonists of 5-HT1A (WAY100135), 5-HT2A (Ketanserin), 5-HT3 (Ondansetron), 5-HT4 (GR125487), and 5-HT7 (SB269970) receptors in the same experimental model. In order to gain further insight into the beneficial action shown by Ketanserin, we tested its ability to prevent colonic apoptosis, typically increased in clinical and experimental colitis and regarded as contributing factor in IBD pathogenesis (Günther et al., 2013), and to promote epithelial cell protection via the stress-inducible HSP70, whose expression has been reported to be down-regulated in IBD (Hu et al., 2007). Finally, given the massive leukocyte recruitment triggered by blockade of 5-HT1A receptors in colitic animals, we investigated the potential protective activity afforded by their stimulation through the administration of the selective 5-HT1A agonist 8-OH-DPAT.
Materials and Methods
Swiss CD1 female mice, 6–10 weeks old (Charles River Laboratories, Calco, Italy) weighing 25–30 g, were housed five per cage, under standard conditions (12:12 h light–dark cycle, water and food ad libitum, 22–24°C). All the experiments were performed in accordance with Guiding Principles in the Care and Use of Animals (DL116/92) and were authorized by the local Animal Care Committee "Organismo preposto al benessere degli animali" and by Italian Ministry of Health.
Induction of Colitis and Experimental Protocol
To induce colitis, mice were lightly anesthetized with diethyl ether and a 10-cm long PE-50 tubing attached to a tuberculin syringe was inserted 2 cm into the colon; 50 μl of a 10% (w/v) TNBS solution in 50% ethanol was administered to mice, maintained in a vertical head-down position for 1 min to avoid leakage of the haptenating agent. Normal (N) animals, receiving 50 μl saline (0.9% NaCl) intrarectally, were included in the study as reference physiological group.
Mice were randomly assigned to the N group (16 animals), receiving subcutaneously saline (10 ml/kg), or to the following experimental groups of colitic animals: saline (C, 10 ml/kg; 16 animals), WAY100135 (W, 5 mg/kg; 12 animals), Ketanserin (K, 5 mg/kg; 12 animals), Ondansetron (O, 10 mg/kg; 12 animals), GR125487 (G, 10 mg/kg; 12 animals), SB269970 (S, 10 mg/kg; 12 animals) and 8-OH-DPAT (OH, 1 mg/kg; 12 animals). The drugs, subcutaneously administered at doses chosen according to our preliminary experiments (data not shown) as effective and not toxic, were repeated twice daily, starting 1 h after TNBS instillation. Animals were daily weighed and wellness status was assessed by unaware observers. Mice were euthanized 3 days after TNBS administration by CO2 inhalation, and organs (colon, liver, lungs) and blood samples were collected for macroscopic, microscopic, and biochemical analyses. In particular, for colon assays, each group of animals was randomly subdivided in three subgroups: colons excised from each subset was reserved either for histological analysis, for MPO activity determination or for cytokines assays. In N, C, W, K, and OH subsets randomly assigned to cytokines determination, colons were longitudinally sectioned in two halves, each one being reserved for cytokines determination or, respectively, for 5-HT and SP levels quantification (N, C, W, and OH subgroups) or for Caspase-3 and HSP70 assays (N, C and K subgroups).
Evaluation of Inflammation
Disease Activity Index
Disease activity index is an evaluation of the pathology severity, based on the daily assignment of a score according to Cooper's modified method (Cooper et al., 1993), on the basis of body weight loss, stool consistency, and rectal bleeding. The scores were quantified as follows: stool consistency: 0 (normal), 1 (soft), 2 (liquid); body weight loss: 0 (<5%), 1 (5–10%), 2 (10–15%), 3 (15–20%), 4 (20–25%), 5 (>25%) and rectal bleeding: 0 (absent), 1 (present). Mice were scored blindly by two investigators.
Macroscopic Score (MS)
After the euthanasia, the colon was removed and opened along the mesenteric line, gently flushed with saline solution and immediately evaluated for the presence of colonic inflammation. MS was quantified by a blinded investigator, according to previously published criteria (Wallace et al., 1989; Khan et al., 2002), as the sum of scores (max = 12) attributed to the presence of points of stenosis and hypertrophic zones (0, absent; 1, 1 stenosis; 2, 2 stenoses; 3, more than 2 stenoses), mucus (0, absent; 1, present), adhesions (0, absent; 1, 1 adhesion between colon and other intra-abdominal organs; 2, 2 adhesions; 3, more than 2 adhesions), intraluminal hemorrhage (0, absent; 1, present), erythema (0, absent; 1, area < 1 cm2; 2, area > 1 cm2), ulceration and necrosis zones (0, absent; 1, area < 1 cm2; 2, area > 1 cm2).
Colon Length and Thickness
To evaluate muscular contraction and deposition of amorphous material induced by massive inflammation, the length of colon, and cecum and their weight were measured, while weight/length ratio was calculated to estimate colon thickness (Bischoff et al., 2009).
Colonic and Hepatic Edema
Colon and liver edemas were measured according to Moore–Olufemi modified method (Moore-Olufemi et al., 2005). After euthanasia, organs were rinsed with saline solution, gently milked dry, and 1 cm long colon segments and a portion of liver were cut and weighed immediately (wet weight, WW). Tissues were then allowed to dry for 72 h at room temperature. Dry weights (DW) were measured and used to determine tissue fluid content from the following wet to dry ratio: (WW–DW)/DW.
Colonic and Pulmonary MPO Activity Assay
Myeloperoxidase activity, an indicator of tissue neutrophil accumulation, was determined according to Krawisz's modified method (Krawisz et al., 1984). After being weighed, each colonic and lung sample was homogenized in ice-cold potassium phosphate buffer (100 mM, pH 7.4) containing aprotinin 1 μg/ml (1:10, v/v) and centrifuged for 20 min at 12500 RCF at 4°C. Pellets were re-homogenized in five volumes of ice-cold potassium phosphate buffer (50 mM, pH 6) containing 0.5% hexadecylthrimethyl-ammoniumbromide and aprotinin 1 μg ml-1. The samples were subjected to three cycles of freezing and thawing, and then centrifuged for 30 min at 15500 RCF at 4°C. 100 microliter of the supernatant was then allowed to react with 900 μl of a buffer solution containing o-dianisidine (0.167 mg ml-1) and 0.0005% H2O2. Each assay was performed in duplicate and the rate of change in absorbance was measured spectrophotometrically at 470 nm (Jenway, mod. 6300, Dunmow, Essex, England). The sensitivity of the assay was 10 mU/ml, 1 unit of MPO being defined as the quantity of enzyme degrading 1 μmol of peroxide per minute at 25°C. Data were normalized with edema values and expressed as U/g of dry weight tissue.
Colonic Cytokines Levels
After euthanasia, colon segments were homogenized for 1 min in 700 μl of tissue lysis buffer containing 20 mM Tris, 150 mM NaCl, 1% Nonidet P-40, 0.5% sodium deoxycholate, 1 mM EDTA and 0.1% SDS (pH 7.5) and protease inhibitors cocktail (pepstatin, aprotinin, and leupeptin 1 μg ml-1). Samples were then centrifuged for 20 min at 16500 RCF at 4°C and the supernatant was collected. Total protein concentration was quantified using Pierce BCA protein assay kit (ThermoFisher Scientific Inc., MA, USA) and each sample diluted to 5 mg/ml. IL-1β, TNFα, IFNγ, and IL-10 colonic concentrations were then determined in duplicate in 100 μl samples, using commercially available ELISA kits (PRODOTTI GIANNI S.p.A., Milano, Italy), according to the manufacturer's protocol, with assays sensitivity, respectively, of 1 pg/ml (IL-1β), 8 pg/ml (TNFα), 10 pg/ml (IFNγ), and 30 pg/ml (IL-10). Results were expressed as pg/mg protein.
Plasmatic Cytokines Levels
Blood was collected by cardiac puncture after CO2 euthanasia, using heparin as anticoagulant (150 IU/ml, 1:9 v/v), and centrifuged at 6500 RCF for 15 min to obtain plasma. Plasma samples were stored at –20°C until the day of the assay. IL-1β and TNFα plasmatic concentrations were determined in duplicate using commercially available ELISA kits according to the procedures supplied by the manufacturer (PRODOTTI GIANNI S.p.A., Milano, Italy) and the results expressed as pg/ml of plasma.
Colonic 5-HT Levels
5-HT concentration was determined in the supernatant of colon homogenates with a commercially available EIA kit (Abnova) according to the manufacturer's protocol. Samples were weighed, homogenized for 1 min in 1 ml of ice-cold 0.01 M HCl solution containing 1% ascorbic acid and centrifuged at 4000 RCF at 4°C for 15 min. Enzymatic reaction products were determined in duplicate spectrophotometrically at 450 nm, the assay sensitivity being 5 ng/ml (Jenway, mod. 6300, Dunmow, Essex, England). Results were normalized with colonic edema values and expressed as μ/g dry weight tissue.
Colonic SP Levels
Substance P concentration was determined in the supernatant of colon homogenates with a commercially available EIA kit (Cayman Chemicals) following manufacturer's protocol. Samples were weighed, homogenized for 1 min in 1 ml of ice-cold PBS (pH 7.4) solution containing protease inhibitors cocktail (pepstatin, aprotinin, and leupeptin 1 μg ml-1) and centrifuged at 6500 RCF at 4°C for 15 min. Enzymatic reaction products were determined in duplicate by spectrophotometry at 405 nm, the assay sensitivity being 8 pg/ml. Results were expressed as ng/g dry weight tissue.
Plasmatic Nitrites Levels
Nitrates plus nitrites levels in plasma samples were determined according to Guevara's method for the determination of nitrite/nitrate by Griess reaction (Guevara et al., 1998). 100 microliter of plasma were incubated with 10 μl of nitrate reductase (500 mU/ml) and 10 μl of NADPH (860 μM) for 2 h at 37°C, in the dark, to reduce nitrates to nitrites. After reduction, 240 μl of methanol/diethyl ether solution (3:1 v/v) were added to each sample and incubated overnight to induce protein denaturation and precipitation. The following day, samples were centrifuged for 10 min at 12500 RCF at 4°C, 100 μl of the supernatant were mixed with 25 μl of HCl 6N and 25 μl of 37.5 M sulphanilic acid and incubated for 10 min at room temperature. Finally, 25 μl of 12.5 M 1-naftilendiammine were added to each sample and incubated for 30 min at room temperature. The absorbance of the samples were measured in duplicate by spectrophotometry at 550 nm and expressed as μg/ml of plasma, the assay sensitivity being 100 ng/ml.
Colonic Caspase-3 and HSP70 Levels
Samples of colon were homogenized for 1 min in 1 ml of ice-cold PBS (pH 7.4) solution containing protease inhibitors cocktail (pepstatin, aprotinin, and leupeptin 1 μg ml-1) and centrifuged at 3000 RCF at 4°C for 15 min. The supernatant was collected, total protein concentration was quantified using Pierce BCA protein assay kit (ThermoFisher Scientific Inc., MA, USA) and an aliquot of the supernatant containing 100 mg proteins/ml (for Caspase-3 assay) and 200 mg proteins/ml (for HSP70 assay) were tested in commercially available ELISA kits (PRODOTTI GIANNI S.p.A., Milano, Italy), according to the manufacturer's protocols. Each sample was assayed in duplicate and the optical density of the final enzymatic reaction products was determined at 450 nm. The sensitivity of the assays was 0.156 ng/ml (Caspase-3) and 200 pg/ml (HSP70). Results were expressed as ng/mg protein (Caspase-3) or as pg/mg protein (HSP70).
Intestinal Histology
Samples of colon harvested from normal animals and from colitic mice administered with saline, Ketanserin 5 mg/kg and 8-OH-DPAT 1 mg/kg were flushed with saline, immersion-fixed in 10% neutral buffered formalin overnight, dehydrated and embedded in paraffin. For each sample, at least five transverse 5-μm sections were cut both in the proximal and in the distal colon, stained with hematoxilin-eosin and examined in a light microscope (Nikon Eclipse E800) by a person unaware of the treatment. To assess histological injury in each section, the severity of mucosal destruction (0, normal; 1, mild; 2, moderate; 3, severe) and the extent of leukocytes infiltration in the lamina propria and submucosa (0, absent; 1, mild; 2, pronounced) were graded and the sum of scores was calculated (maximum score: 7) (modified from Bischoff et al., 2009). Since histological injury produced by TNBS instillation was localized almost exclusively in the distal portion of the colon, the average value of histological score was determined only from sections of distal colon, pooled with those determined for the other animals of the same experimental group and the mean value ± SEM was calculated.
Statistical Analysis
All data were presented as means ± SEM. Comparison among groups were made using analysis of variance (one-way ANOVA) followed by Bonferroni's post-test. Non-parametric Kruskal-Wallis analysis followed by Dunn's post-test was applied for statistical comparison of DAI, MS, and cytokines levels. P < 0.05, P < 0.01, and P < 0.001 showed, respectively, statistically significant, highly significant, or extremely highly significant differences. All analyses were performed using Prism 4 software (GraphPad Software Inc. San Diego, CA, USA).
WAY100135 (5-HT1A antagonist), GR125487 (5-HT4 anta gonist), and SB-269970 (5-HT7 antagonist) were purchased from Tocris Bioscence (Bristol, UK). Ondansetron (5-HT3 antagonist) was purchased from Glaxo Wellcome (Uxbridge, Middlesex, UK). 8-OH-DPAT (5-HT1A agonist), Ketanserin (5-HT2A antagonist), TNBS, MPO and all the other chemicals of reagent grade were purchased from Sigma–Aldrich Chemical Company (St. Louis, MO, USA). Drugs were dissolved in saline solution the day of the experiment.
TNBS Induced Severe Colitis in Mice
Intrarectal administration of the haptenating agent TNBS in saline-treated mice induced a global worsening of health conditions, expressed as DAI index, with respect to N animals: progressive reduction of body weight and loss of stool consistency were evident and reached their maximum at day 4, when animals were sacrificed (P < 0.001 Dunn's test, Figure 1A). Macroscopic damage scores, based on the presence of adhesions, points of stenosis, mucus, erythemas, and ulcers in colon specimens were significantly increased after induction of TNBS colitis compared with N animals, that had only sporadically and scarcely visible rectal erythemas (P < 0.001 Dunn's test, Figure 1B); colonic length was markedly reduced (P < 0.001 Bonferroni's test, Figure 1C) and, conversely, thickness (P < 0.001 Bonferroni's test, Figure 1D) and edema (5.3 ± 0.1 C vs. 3.6 ± 0.3 N group, P < 0.001 Bonferroni's test) augmented following TNBS administration.
FIGURE 1. Effects of 5-HT receptor antagonists on TNBS-induced disease severity. DAI at day 4 (A), MS (B), colonic length (C), and colonic thickness (D) assessed in normal mice (N) and in TNBS-treated mice administered with vehicle (C), WAY100135 5 mg/kg (W), Ketanserin 5 mg/kg (K), Ondansetron 10mg/kg (O), GR125487 10 mg/kg (G) and SB269970 10 mg/kg (S) (n = 6–12 data per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. N mice; #P < 0.05, ##P < 0.01 vs. C mice; one-way ANOVA followed by Bonferroni's post-test. Kruskal–Wallis analysis followed by Dunn's post-test was applied for statistical comparison of DAI and MS.
Consistent with these alterations, the microscopic analysis revealed diffuse epithelial degeneration and massive neutrophilic infiltration of the distal colonic wall in TNBS-inoculated mice (Figure 2B; histological score: 6.0 ± 1.0) with respect to normal animals (Figure 2A; histological score: 0).
FIGURE 2. Histology. Representative hematoxylin-eosin stained sections of colonic specimens harvested from normal mice (A) and from TNBS-treated mice administered with vehicle (B), Ketanserin 5 mg/kg (C) or 8-OH-DPAT 1 mg/kg (D). TNBS colonic instillation caused epithelial degeneration, neutrophilic infiltration, and submucosal edema (indicated by arrows) in vehicle-treated animals (B), not overtly modified either by Ketanserin (C) or 8-OH-DPAT (D) treatment.
These local morphological changes were accompanied by systemic inflammatory responses, represented by increased liver edema (2.20 ± 0.02 vs. 1.84 ± 0.02 N group, P < 0.001 Bonferroni's test), intense infiltration of leukocytes in the colon as well as in lungs, witnessed by the increase in MPO activity (P < 0.001 vs. N Bonferroni's test, Figure 3), and by the remarkable up-regulation of pro- and anti-inflammatory cytokines both in colonic tissues and in plasma of colitic mice (Figure 4). In this condition of severe colonic inflammation, tissue 5-HT content (P < 0.001 Bonferroni's test) and plasmatic nitrites (P < 0.05 Bonferroni's test) were more than doubled in C mice compared to N animals (Table 1).
FIGURE 3. Effects of 5-HT receptor antagonists on TNBS-induced neutrophil infiltration in colon and lung. MPO activity in colonic (A) and lung (B) tissues excised from normal mice (N) and from TNBS-treated mice administered with vehicle (C), WAY100135 5mg/kg (W), Ketanserin 5mg/kg (K), Ondansetron 10mg/kg (O), GR125487 10mg/kg (G), and SB269970 10mg/kg (S) (n = 6–12 data per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. N mice; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. C mice; one-way ANOVA followed by Bonferroni's post-test.
FIGURE 4. Effects of 5-HT receptor antagonists on cytokines levels. Colonic concentrations of TNFα (A), IL-1β (B), IFNγ (C), and IL-10 (D) and plasmatic levels of TNFα (E) and IL-1β (F) in normal mice (N) and in TNBS-treated mice administered with vehicle (C), WAY100135 5 mg/kg (W), Ketanserin 5 mg/kg (K), Ondansetron 10 mg/kg (O), GR125487 10 mg/kg (G) and SB269970 10 mg/kg (S) (n = 6–12 data per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. N mice; #P < 0.05 vs. C mice; Kruskal–Wallis analysis followed by Dunn's post-test.
TABLE 1. 5-HT, nitrites, and SP levels in plasma and colonic samples excised from normal mice (N) and from colitic mice administered with saline (C), WAY100135 5 mg/kg (W), and 8-OH-DPAT 1 mg/kg (OH) (n = 6–12 data per group).
5-HT2A Blocker Attenuated the Severity of TNBS-Induced Colitis
Among the different 5-HT receptor antagonists tested, only repeated administration of 5-HT2A antagonist Ketanserin 5 mg/kg slightly improved health conditions of colitic mice and counteracted both colon shortening (P < 0.05) and thickening (P < 0.01) produced by TNBS intrarectal instillation, even if without reducing the MS (Figure 1) or the microscopic damage (Figure 2C; histological score: 6.3 ± 0.7).
Blockade of 5-HT2A receptors significantly contrasted the infiltration of the TNBS-inflamed intestine with polymorphonuclear leukocytes (P < 0.05) and dampened the increase in lung MPO activity exhibited by colitic mice (Figure 3). The anti-inflammatory action of Ketanserin was evident also when considering the levels of bowel and plasmatic cytokines: indeed, the 5-HT2A blocker was able to globally revert the increase in inflammatory cytokines evoked in TNBS colitis both in colon and in plasma (Figure 4), where TNFα and IL-1β concentrations were reduced, respectively, up to 5 (P < 0.05) and 2.5 times compared to C animals.
In order to investigate the possible mechanism underlying the protective effects displayed by Ketanserin, colonic levels of caspase-3, apoptotic marker, and of HSP70, a stress-inducible protein with cytoprotective properties, were determined in N, C, and Ketanserin-treated mice. As expected, TNBS enema produced a remarkable increase of caspase-3 levels and a highly significant reduction of HSP70: Ketanserin was able to counteract caspase-3 rise (P < 0.05 vs. C) but it did not affect HSP70 decrease (Figure 5).
FIGURE 5. Effect of Ketanserin on colonic caspase-3 and HSP70 levels. Colonic concentrations of caspase-3 (A) and HSP70 (B) in normal mice (N) and in TNBS-treated mice administered with vehicle (C) or Ketanserin 5 mg/kg (K) (n = 6–12 data per group). ∗P < 0.05, ∗∗P < 0.01 vs. N mice; #P < 0.05 vs. C mice; one-way ANOVA followed by Bonferroni's post-test.
5-HT1A Blocker Worsened Neutrophil Infiltration in TNBS-Induced Colitis
Subcutaneous administration of 5-HT1A antagonist WAY100135 10 mg/kg to TNBS-treated mice did not modify significantly the progression of the pathology nor the macroscopic damage score, slightly increasing only colon wall thickness in comparison with C mice (Figure 1). However, both intestinal and lung tissues excised from animals administered with 5-HT1A blocker exhibited a conspicuous up-regulation of MPO activity, an index of neutrophil recruitment, with respect to vehicle-treated mice: in fact, treatment with WAY100135 nearly doubled MPO levels from 96.7 ± 7.3 U/g in the gut and 116.6 ± 6.9 U/g in lungs to, respectively, 150.8 ± 20.3 U/g and 182.6 ± 19.7 U/g (Figure 3). 5-HT1A antagonist did not overtly modify the increased amount either of local or systemic inflammatory cytokines produced by TNBS (Figure 4).
5-HT1A Agonist Weakly Mitigated the Severity of TNBS-Induced Colitis
Administration of 5-HT1A agonist 8-OH-DPAT 1 mg/kg to TNBS-treated mice remarkably improved their health conditions, DAI scoring being significantly lower than that of vehicle-treated mice every day of the observation period (Figure 6). Moreover, exogenous stimulation of 5-HT1A receptors attenuated the thickening (55.0 ± 2.4 mg/cm vs 68.7 ± 3.4 C mice, P < 0.01) and the increase in colonic edema (4.8 ± 0.2 vs. 5.4 ± 0.1 C mice, P < 0.05) observed in segments excised from control mice, but did not reduce the mucosal damage, macroscopically or histo-pathologically detected (Figure 2D; histological score: 6.0 ± 1.0), nor the local and systemic leukocyte infiltration (data not shown) or cytokines release (Figure 7).
FIGURE 6. Effects of 5-HT1A ligands on disease progression. Colitis severity, expressed as DAI, assessed on all four days following TNBS i.r. instillation in normal mice (black circles) and in mice administered with vehicle (black squares), WAY100135 5 mg/kg (black triangles) and 8-OH-DPAT 1mg/kg (white triangles) (n = 6–12 data per group). ∗∗P < 0.01, ∗∗∗P < 0.001 vs. N mice; #P < 0.05, ###P < 0.001 vs. C mice; Kruskal–Wallis analysis followed by Dunn's post-test.
FIGURE 7. Effects of 8-OH-DPAT on cytokines levels. Colonic concentrations (pg/mg protein) of TNFα, IL-1β, IFNγ, and IL-10 and plasmatic levels (pg/ml) of TNFα and IL-1β in normal mice (white bars) and in TNBS-treated mice administered with vehicle (black bars) and 8-OH-DPAT 1mg/kg (gray bars) (n = 6–12 data per group). ∗P < 0.05, ∗∗P < 0.01 vs. N mice; Kruskal–Wallis analysis followed by Dunn's post-test.
Interestingly, bowel 5-HT content was significantly decreased following 8-OH-DPAT treatment with respect to vehicle-treated colitic animals (P < 0.05, Table 1). Along with 5-HT, we investigated the role of 5-HT1A receptors in affecting the release also of other mediators typically involved in intestinal inflammation. Accordingly, the levels of plasmatic nitrites and of colonic SP were assessed following 5-HT1A stimulation and blockade in TNBS-treated mice: while 5-HT1A antagonist significantly augmented the levels of nitrites when compared with N (P < 0.001) or C mice (P < 0.05), 8-OH-DPAT did not modify either nitrites or SP levels (Table 1).
Blockade of 5-HT3, 5-HT4, and 5-HT7 Receptors Did Not Remarkably Affect the Severity of TNBS-Induced Colitis
The administration of 5-HT4 antagonist GR125487 10 mg/kg reduced DAI index at day 4 and colon thickness (P < 0.05) with respect to C mice (Figure 1), but had no clear effect on the other local and systemic inflammatory responses. Blockade of 5-HT3 receptors by Ondansetron 10 mg/kg and 5-HT7 receptors by SB269970 10 mg/kg provoked an increase in mortality rate of, respectively, 40 and 25% (data not shown), preventing experiments at higher dosages. Neither of the treatments applied were able to significantly aggravate or improve any of the clinical, macroscopic and biochemical parameters examined.
The results collected in the present study indicate that:
• TNBS intrarectal instillation in mice produced a strong local and systemic inflammatory response, associated with increased colonic levels of 5-HT;
• improved global health conditions, reduced morphological colonic changes and down-regulation of neutrophil recruitment and of inflammatory cytokines levels were produced by antagonism of 5-HT2A receptors;
• blockade of 5-HT1A receptors worsened neutrophil recruitment while, conversely, exogenous stimulation of the same receptor subtype delayed and mitigated the severity of TNBS-induced colitis, contrasting the increase in colonic 5-HT content;
• blockade of 5-HT3, 5-HT4, and 5-HT7 receptors did not remarkably affect the progression and outcome of the pathology or only slightly improved it.
The central role of 5-HT in modulating GI peristaltic and secretory reflexes has been known for a long time (Tonini, 2005); more recently, the influence of 5-HT also on innate and adaptive immune responses and on enteric inflammation has been emerging. The clinical and experimental studies currently available indicate a pro-inflammatory contribution of the monoamine to the pathogenesis of intestinal disorders like IBD (El-Salhy et al., 1997; Bischoff et al., 2009; Ghia et al., 2009; Margolis et al., 2014), although the mechanisms underlying and the receptor subtypes mediating this effect have yet to be identified.
In order to shed some light on the role played in intestinal inflammation by the 5-HT receptor subtypes most abundantly expressed in the GI tract, we chose a conventional chemical model of experimental colitis mainly driven by a Th1-mediated immune response, although well aware that no single model can perfectly mimic human IBD complexity (Kiesler et al., 2015). As expected, intrarectal TNBS challenge in mice produced a severe state of illness accompanied by colonic and systemic inflammatory responses and, consistent with the literature, by a higher 5-HT tissue amount (Oshima et al., 1999; Linden et al., 2003). Therefore, our findings apparently represent a new clue in favor of the pro-inflammatory role of the monoamine. However, when the overall results of the present investigation were examined, a more complex picture emerged, actually mirroring a multifaceted role of endogenous 5-HT in intestinal inflammation.
Repeated treatment of TNBS-challenged mice with different 5-HT receptor antagonists produced heterogeneous results, showing that endogenous 5-HT could provide simultaneously a deleterious and, alternatively, a beneficial contribution. Among the various 5-HT blockers applied, only 5-HT2A antagonist Ketanserin protected animals against TNBS-induced colitis, attenuating the gravity of the local alterations and of the systemic inflammatory state. The ability of Ketanserin to markedly prevent both the colon shortening and the development of an inflammatory infiltrate, consisting presumably mainly of polymorphonuclear leukocytes, seems to indicate that, once released by EC cells following TNBS challenge (Bischoff et al., 2009; Margolis et al., 2014), endogenous 5-HT may participate in the up-regulation of colonic and plasmatic inflammatory cytokines and, consequently, to the recruitment of activated phagocytes in the bowel through the stimulation of 5-HT2A receptors. 5-HT2 receptors have been described to influence several immune responses, by promoting the chemotaxis of immature dendritic cells (Muller et al., 2009) and eosinophils (Boehme et al., 2004) and activating CD4+ and CD8+ T lymphocytes (Inoue et al., 2011); conversely, blockade of the same receptor subtype by Ketanserin was able to counteract the up-regulation of inflammatory cytokines in a model of systemic inflammation (Sugino et al., 2009). Our present findings are in line with these observations and point toward the beneficial immunomodulatory effects of selective 5-HT2A pharmacological blockade.
Besides the reduction in pro-inflammatory cytokines levels, the amelioration induced by Ketanserin on TNBS colitis may depend also on its anti-apoptotic effects on colonic tissues. Indeed, intestinal epithelial cells apoptosis is increased in the mucosa of IBD patients and in experimental colitis, although it is still unclear whether excessive cell death is the origin or simply a consequence of the inflammatory environment promoted in IBD (Günther et al., 2013); as a result, anti-apoptotic effects are often considered decisive to the protection afforded by a number of different therapeutic strategies in experimental colitis (see for instance Crespo et al., 2012; Arumugam et al., 2015; Yang et al., 2015). Interestingly, beneficial anti-apoptotic effects were demonstrated also by 5-HT2 antagonists in myocardial injury (Rajesh et al., 2006; Bharti et al., 2015). Altogether, our data lend support to a potential application of 5-HT2A antagonists as supplemental strategy, combining anti-inflammatory and anti-apoptotic actions, for the treatment of intestinal inflammatory disorders.
Blockade of 5-HT4 receptors by GR125487 only partially overlapped the protective action of Ketanserin, weakly improving clinical conditions and attenuating colonic thickening: it is likely that the localization of 5-HT4 receptors on dendritic cells and their activating effects on immune cells functions (Idzko et al., 2004) could come into play in this regard.
If, in our experimental conditions, endogenous activation of 5-HT2A and 5-HT4 receptors seems to contribute to the TNBS-induced damage, simultaneous stimulation of 5-HT1A sites apparently contrasts the neutrophil recruitment triggered both locally and systemically by TNBS challenge, as demonstrated by the enhancing effect shown by 5-HT1A antagonist WAY100135 on the strong leukocytes migration in colon and lungs induced by the instillation of the haptenating agent. Consistent with these findings is the protective action displayed by 5-HT1A agonist 8-OH-DPAT, which improved the general health conditions of colitic mice and attenuated the wall structural alterations elicited by TNBS application. This advantageous effect exerted by stimulation of 5-HT1A sites is not easily interpreted by taking into account their immunomodulatory actions documented in vitro, such as the inhibition of T cell proliferation and activity produced by 5-HT1A antagonism (Aune et al., 1994). On the other hand, it is well known that 5-HT1A receptors are present also on neural elements in the gut, where they pre-synaptically inhibit the release of Ach and of non-cholinergic neurotransmitters (LePard et al., 2004). Following this line of evidence, we tested the hypothesis that activation of 5-HT1A receptors could interfere with the release of SP, a neuropeptide documented to be up-regulated and directly involved in IBD-induced intestinal inflammation (Gross and Pothoulakis, 2007). On the basis of the findings here collected we could rule out this possible mechanism of action, since neither 5-HT1A agonist nor antagonist produced significant changes in the levels of colonic SP.
As regards the effects of 5-HT1A receptor modulation on NO availability, several investigations documented a greatly increased production of this labile mediator in IBD and in experimental colitis and highlighted its pro-inflammatory and detrimental role or vice-versa its homeostatic and protective action in the pathogenesis of intestinal injury (Perner and Rask-Madsen, 1999; Kolios et al., 2004). In our conditions, the increased levels of plasmatic nitrites, stable metabolites of NO, detected in TNBS-colitis seem to support the potential harmful effects of this mediator; blockade of 5-HT1A receptors further augmented the amount of NO, likely contributing to the negative outcome of WAY100135 treatment. On the whole, we can speculate that endogenous 5-HT may participate via 5-HT1A receptors in limiting the massive neutrophils recruitment triggered in TNBS-colitis possibly by restraining NO over-production.
Interestingly, 5-HT1A receptors have been localized also on EC cells (Kirchgessner et al., 1996) from which 5-HT can modulate its own release (Schworer and Ramadori, 1998). The efficacy exhibited by 8-OH-DPAT in reducing 5-HT colonic content could therefore reflect the role played by 5-HT1A receptors in regulating 5-HT availability from EC cells in the inflamed gut. It is tempting to hypothesize that the beneficial effect provided by 5-HT1A agonist in this model of TNBS-induced colitis might depend on the reduced intestinal 5-HT content, further highlighting the pro-inflammatory and deleterious role of the monoamine.
Finally and unexpectedly, neither blockade of 5-HT3 nor that of 5-HT7 receptors was able to significantly modify the disease progression and the intensity of the inflammatory response elicited by TNBS challenge. In fact, our results apparently contrast with recent investigations documenting the protection provided by 5-HT3 antagonists in TNBS-induced colitis in rats (Motavallian et al., 2013) and in 5-fluorouracil-induced intestinal mucositis in mice (Yasuda et al., 2013) and the controversial role, either protective or deleterious, described for 5-HT7 receptors in DSS-induced colitis (Kim et al., 2013; Guseva et al., 2014). However, differences between species, models of colitis and doses of SB-269970 administered could account for the observed discrepancies.
Our findings shed new light on the double-faced role of 5-HT in intestinal delayed inflammation: the prevailing deleterious contribution mediated by 5-HT2A and 5-HT4 receptors activation co-exists and is partially counteracted by 5-HT1A stimulation, suggesting that agents targeting serotoninergic system could be beneficial as an additional therapeutic strategy in IBD only by carefully dissecting the monoamine antithetical effects.
Author Contributions
AR carried out the studies and data analyses. SB performed the statistical analysis, interpreted the data, and wrote the manuscript. VA, FS, VV, and AG participated in performing the experiments and acquiring the data. AC carried out the histological analyses. EB conceived the study and supervised the investigation. All the authors revised the manuscript critically, approved its final version, and agreed to be accountable for all aspects of the work.
This work was supported by a local grant of University of Parma.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Preliminary data were presented at Digestive Disease Week 2013, 18-21 May 2013, Orlando, Florida (USA). The authors wish to thank Dr. Giuseppe Domenichini for his skilful technical assistance.
5-HT, Serotonin; 8-OH-DPAT, 8-Hydroxy-2-(di-n-propylamino)tetralin; C, Control; CD, Crohn's Disease; DAI, Disease Activity Index; EC, Enterochromaffin; G, GR125487; GI, Gastrointestinal; HSP70, Heat Shock Protein-70; IBD, Inflammatory Bowel Disease; IBS, Irritable Bowel Sindrome; IFN, Interferon; IL, Interleukin; K, Ketanserin; MS, Macroscopic Score; MPO, Myeloperoxidase; N, Normal; NO, Nitric oxide; O, Ondansetron; S, SB269970; SERT, Serotonin Reuptake Transporter; SP, Substance P; TNBS, 2,4,6-TriNitroBenzene Sulfonic acid; TNF, Tumor necrosis factor; Tph1, Tryptophan hydroxylase 1; UC, Ulcerative Colitis; W, WAY100135.
Abraham, C., and Cho, J. H. (2009). Inflammatory bowel disease. N. Engl. J. Med. 361, 2066–2078. doi: 10.1056/NEJMra0804647
Arreola, R., Becerril-Villanueva, E., Cruz-Fuentes, C., Velasco-Velazquez, M. A., Garcés-Alvarez, M. E., Hurtado-Alvarado, G., et al. (2015). Immunomodulatory effects mediated by serotonin. J. Immunol. Res. 2015, 354957. doi: 10.1155/2015/354957
Arumugam, S., Sreedhar, R., Thandavarayan, R. A., Giridharan, V. V., Karuppagounder, V., Pitchaimani, V., et al. (2015). Telmisartan treatment targets inflammatory cytokines to suppress the pathogenesis of acute colitis induced by dextran sulphate sodium. Cytokine 74, 305–312. doi: 10.1016/j.cyto.2015.03.017
Aune, T. M., Golden, H. W., and McGrath, K. M. (1994). Inhibitors of serotonin synthesis and antagonists of serotonin 1A receptors inhibit T lymphocyte function in vitro and cell-mediated immunity in vivo. J. Immunol. 153, 489–498.
Belai, A., Boulos, P. B., Robson, T., and Burnstock, G. (1997). Neurochemical coding in the small intestine of patients with Crohn's disease. Gut 40, 767–774. doi: 10.1136/gut.40.6.767
Bertoni, S., Arcaro, V., Vivo, V., Rapalli, A., Tognolini, M., Cantoni, A. M., et al. (2014). Suppression of inflammatory events associated to intestinal ischemia-reperfusion by 5-HT1A blockade in mice. Pharmacol. Res. 81, 17–25. doi: 10.1016/j.phrs.2014.02.002
Bertoni, S., Ghizzardi, P., Cattaruzza, F., Ballabeni, V., Impicciatore, M., Tognolini, M., et al. (2007). Evidence for the involvement of 5-HT2A receptors in mild mesenteric ischemia/reperfusion dysfunctions in mice. Pharmacol. Res. 56, 550–555. doi: 10.1016/j.phrs.2007.10.004
Bharti, S., Rani, N., Bhatia, J., and Arya, D. S. (2015). 5-HT2B receptor blockade attenuates β-adrenergic receptor-stimulated myocardial remodelling in rats via inhibiting apoptosis: role of MAPKs and HSPs. Apoptosis 20, 455–465. doi: 10.1007/s10495-014-1083-z
Bischoff, S. C., Mailer, R., Pabst, O., Weier, G., Sedlik, W., Li, Z., et al. (2009). Role of serotonin in intestinal inflammation: knock out of serotonin reuptake transporter exacerbates 2,4,6-trinitrobenzene sulfonic acid colitis in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 296, G685–G695. doi: 10.1152/ajpgi.90685.2008
Boehme, S. A., Lio, F. M., Sikora, L., Pandit, T. S., Lavrador, K., Rao, S. P., et al. (2004). Cutting edge: serotonin is a chemotactic factor for eosinophils and functions additively with eotaxin. J. Immunol. 173, 3599–3603. doi: 10.4049/jimmunol.173.6.3599
Coleman, N. S., Foley, S., Dunlop, S. P., Wheatcroft, J., Blackshaw, E., Perkins, A. C., et al. (2006). Abnormalities of serotonin metabolism and their relation to symptoms in untreated celiac disease. Clin. Gastroenterol. Hepatol. 4, 874–881. doi: 10.1016/j.cgh.2006.04.017
Cooper, H. S., Murthy, S. N., Shah, R. S., and Sedergran, D. J. (1993). Clinico-pathological study of dextran sulfate sodium experimental murine colitis. Lab. Invest. 69, 238–249.
Crespo, I., San-Miguel, B., Prause, C., Marroni, N., Cuevas, M. J., Gonzalez-Gallego, J., et al. (2012). Glutamine treatment attenuates endoplasmic reticulum stress and apoptosis in TNBS-induced colitis. PLoS ONE 7:e50407. doi: 10.1371/journal.pone.0050407
De Ponti, F. (2004). Pharmacology of serotonin: what a clinician should know. Gut 53, 1520–1535. doi: 10.1136/gut.2003.035568
El-Salhy, M., Danielsson, A., Stenling, R., and Grimelius, L. (1997). Colonic endocrine cells in inflammatory bowel disease. J. Intern. Med. 242, 413–419. doi: 10.1046/j.1365-2796.1997.00237.x
Ghia, J. E., Li, N., Wang, H., Deng, Y., El-Sharkawy, R. T., Coté, F., et al. (2009). Serotonin has a key role in the pathogenesis of experimental colitis. Gastroenterology 137, 1649–1660. doi: 10.1053/j.gastro.2009.08.041
Gross, K. J., and Pothoulakis, C. (2007). Role of neuropeptides in inflammatory bowel disease. Inflamm. Bowel Dis. 13, 918–932. doi: 10.1002/ibd.20129
Guevara, I., Iwanejko, J., Dembińska-Kieć, A., Pankiewicz, J., Wanat, A., Anna, P., et al. (1998). Determination of nitrite/nitrate in human biological material by the simple Griess reaction. Clin. Chim. Acta 274, 177–188. doi: 10.1016/S0009-8981(98)00060-6
Günther, C., Neumann, H., Neurath, M. F., and Becker, C. (2013). Apoptosis, necrosis and necroptosis: cell death regulation in the intestinal epithelium. Gut 62, 1062–1071. doi: 10.1136/gutjnl-2011-301364
Guseva, D., Holst, K., Kaune, B., Meier, M., Keubler, L., Glage, S., et al. (2014). Serotonin 5-HT7 receptor is critically involved in acute and chronic inflammation of the gastrointestinal tract. Inflamm. Bowel Dis. 20, 1516–1529. doi: 10.1097/MIB.0000000000000150
Hu, S., Ciancio, M. J., Lahav, M., Fujiya, M., Lichtenstein, L., Anant, S., et al. (2007). Translational inhibition of colonic epithelial heat shock proteins by IFN-g and TNF-a in intestinal inflammation. Gastroenterology 133, 1893–1904. doi: 10.1053/j.gastro.2007.09.026
Idzko, M., Panther, E., Stratz, C., Muller, T., Bayer, H., Kissel, G., et al. (2004). The serotoninergic receptors of human dendritic cells: identification and coupling to cytokine release. J. Immunol. 172, 6011–6019. doi: 10.4049/jimmunol.172.10.6011
Inoue, M., Okazaki, T., Kitazono, T., Mizushima, M., Omata, M., and Ozaki, S. (2011). Regulation of antigen-specific CTL and Th1 cell activation through 5-hydroxytryptamine 2A receptor. Int. Immunopharmacol. 11, 67–73. doi: 10.1016/j.intimp.2010.10.007
Khan, W. I., Blennerhasset, P. A., Varghese, A. K., Chowdhury, S. K., Omsted, P., Deng, Y., et al. (2002). Intestinal nematode infection ameliorates experimental colitis in mice. Infect. Immun. 70, 5931–5937. doi: 10.1128/IAI.70.11.5931-5937.2002
Kiesler, P., Fuss, I. J., and Strober, W. (2015). Experimental models of inflammatory bowel diseases. Cell. Mol. Gastroenterol. Hepatol. 1, 154–170. doi: 10.1016/j.jcmgh.2015.01.006
Kim, J. J., Bridle, B. W., Ghia, J. E., Wang, H., Syed, S. N., Manocha, M. M., et al. (2013). Targeted inhibition of serotonin type 7 (5-HT7) receptor function modulates immune responses and reduces the severity of intestinal inflammation. J. Immunol. 190, 4795–4804. doi: 10.4049/jimmunol.1201887
Kim, J. J., Wang, H., Terc, J. D., Zambrowicz, B., Yang, Q. M., and Khan, W. I. (2015). Blocking peripheral serotonin synthesis by telotristat etiprate (LX1032/LX1606) reduces severity of both chemical- and infection-induced intestinal inflammation. Am. J. Physiol. Gastrointest. Liver Physiol. 309, G455–G465. doi: 10.1152/ajpgi.00299.2014
Kirchgessner, A. L., Liu, M. T., Raymond, J. R., and Gershon, M. D. (1996). Identification of cells that express 5-hydroxytryptamine1A receptors in the nervous system of the bowel and pancreas. J. Comp. Neurol. 364, 439–455. doi: 10.1002/(SICI)1096-9861(19960115)364:3<439::AID-CNE5>3.0.CO;2-5
Kolios, G., Valatas, V., and Ward, S. G. (2004). Nitric oxide in inflammatory bowel disease: a universal messenger in an unsolved puzzle. Immunology 113, 427–437. doi: 10.1111/j.1365-2567.2004.01984.x
Krawisz, J. E., Sharon, P., and Stenson, W. F. (1984). Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models. Gastroenterology 87, 1344–1350.
LePard, K. J., Ren, J., and Galligan, J. J. (2004). Presynaptic modulation of cholinergic and non-cholinergic fast synaptic transmission in the myenteric plexus of guinea pig ileum. Neurogastroenterol. Motil. 16, 355–364. doi: 10.1111/j.1365-2982.2004.00505.x
Linden, D. R., Chen, J. X., Gershon, M. D., Sharkey, K. A., and Mawe, G. M. (2003). Serotonin availability is increased in mucosa of guinea pigs with TNBS-induced colitis. Am. J. Physiol. Gastrointest. Liver Physiol. 285, G207–G216. doi: 10.1152/ajpgi.00488.2002
Magro, F., Vieira-Coelho, M. A., Fraga, S., Serrao, M. P., Veloso, F. T., Ribeiro, T., et al. (2002). Impaired synthesis or cellular storage of norepinephrine, dopamine, and 5-hydroxytryptamine in human inflammatory bowel disease. Dig. Dis. Sci. 47, 216–224. doi: 10.1023/A:1013256629600
Manocha, M., and Khan, W. I. (2012). Serotonin and GI disorders: an update on clinical and experimental studies. Clin. Transl. Gastroenterol. 3, e13. doi: 10.1038/ctg.2012.8
Margolis, K. G., Stevanovic, K., Li, Z., Yang, Q. M., Oravecz, T., Zambrowicz, B., et al. (2014). Pharmacological reduction of mucosal but not neuronal serotonin opposes inflammation in mouse intestine. Gut 63, 928–937. doi: 10.1136/gutjnl-2013-304901
Moore-Olufemi, S. D., Kozar, R. A., Moore, F. A., Sato, N., Hassoun, H. T., Cox, C. S. Jr., et al. (2005). Ischemic preconditioning protects against gut dysfunction and mucosal injury after ischemia/reperfusion injury. Shock 23, 258–263.
Motavallian, A., Minaiyan, M., Rabbani, M., Andalib, S., and Mahzouni, P. (2013). Involvement of 5-HT3 receptors in anti-inflammatory effects of Tropisetron on experimental TNBS-induced colitis in rat. Bioimpacts 3, 169–176. doi: 10.5681/bi.2013.021
Muller, T., Durk, T., Blumenthal, B., Grimm, M., Cicko, S., Panther, E., et al. (2009). 5-Hydroxytryptamine modulates migration, cytokine and chemokine release and T cell priming capacity of dendritic cells in vitro and in vivo. PLoS ONE 4:e6453. doi: 10.1371/journal.pone.0006453
Oshima, S., Fujimura, M., and Fukimiya, M. (1999). Changes in number of serotonin-containing cells and serotonin levels in the intestinal mucosa of rats with colitis induced by dextran sodium sulfate. Histochem. Cell Biol. 112, 257–263. doi: 10.1007/s004180050445
Perner, A., and Rask-Madsen, J. (1999). Review article: the potential role of nitric oxide in chronic inflammatory bowel disorders. Aliment. Pharmacol. Ther. 13, 135–144. doi: 10.1046/j.1365-2036.1999.00453.x
Rajesh, K. G., Suzuki, R., Maeda, H., Murio, Y., and Sasaguri, S. (2006). 5-HT2 receptor blocker sarpogrelate prevents down-regulation of antiapoptotic protein Bcl-2 and protects the heart against ischemia-reperfusion injury. Life Sci. 79, 1749–1755. doi: 10.1016/j.lfs.2006.06.026
Schworer, H., and Ramadori, G. (1998). Autoreceptors can modulate 5-hydroxytriptamine release from porcine and human small intestine in vitro. Naunyn Schmiedebergs Arch. Pharmacol. 357, 548–552. doi: 10.1007/PL00005206
Spiller, R. (2008). Serotonin and GI clinical disorders. Neuropharmacology 55, 1072–1080. doi: 10.1016/j.neuropharm.2008.07.016
Sugino, H., Futamura, T., Mitsumoto, Y., Maeda, K., and Marunaka, Y. (2009). Atypical antipsychotics suppress production of pro-inflammatory cytokines and up-regulate interleukin-10 in lipopolysaccharide-treated mice. Prog. Neuropsychopharmacol. Biol. Psychiatry 33, 303–307. doi: 10.1016/j.pnpbp.2008.12.006
Teramoto, Y., Urano, T., Nagai, N., Takada, Y., Ikeda, K., and Takada, A. (1998). Plasma levels of 5-HT and 5-HIAA increased after intestinal ischemia/reperfusion injury. Jpn. J. Physiol. 48, 333–339. doi: 10.2170/jjphysiol.48.333
Tonini, M. (2005). 5-Hydroxytryptamine effects in the gut: the 3, 4 and 7 receptors. Neurogastroenterol. Motil. 17, 637–642. doi: 10.1111/j.1365-2982.2005.00716.x
Wallace, J. L., MacNaughton, W., Morris, G. P., and Beck, P. L. (1989). Inhibition of leukotriene synthesis markedly accelerates healing in a rat model of inflammatory bowel disease. Gastroenterology 96, 29–36.
Yang, J., Liu, X. X., Fan, H., Tang, Q., Shou, Z. X., Zuo, D. M., et al. (2015). Extracellular vesicles derived from bone marrow mesenchimal stem cells protect against experimental colitis via attenuating colon inflammation, oxidative stress and apoptosis. PLoS ONE 10:e0140551. doi: 10.1371/journal.pone.0140551
Yasuda, M., Kato, S., Yamanaka, N., Iimori, M., Matsumoto, K., Utsumi, D., et al. (2013). 5-HT3 receptor antagonists ameliorate 5-fluorouracil-induced intestinal mucositis by suppression of apoptosis in murine intestinal crypt cells. Br. J. Pharmacol. 168, 1388–1400. doi: 10.1111/bph.12019
Keywords: intestine, inflammation, 5-HT2A receptor, 5-HT1A receptor, apoptosis
Citation: Rapalli A, Bertoni S, Arcaro V, Saccani F, Grandi A, Vivo V, Cantoni AM and Barocelli E (2016) Dual Role of Endogenous Serotonin in 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis. Front. Pharmacol. 7:68. doi: 10.3389/fphar.2016.00068
Received: 25 January 2016; Accepted: 08 March 2016;
Published: 22 March 2016.
Edited by:David Bulmer, Barts and The London School of Medicine and Dentistry, UK
Reviewed by:Anthony C. Johnson, University of Oklahoma Health Sciences Center, USA
Silvia Melgar, University College Cork, Ireland
Copyright © 2016 Rapalli, Bertoni, Arcaro, Saccani, Grandi, Vivo, Cantoni and Barocelli. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Simona Bertoni, firstname.lastname@example.com


text:
A brand new Boeing 737 MAX 8 plane delivered to Ethiopian Airlines just four months ago and carrying 157 people, has crashed after taking off from Addis Ababa. There are no survivors among 157 people on board an Ethiopian Airlines Boeing 737 that crashed en route from Addis Ababa to Nairobi, Ethiopia's state broadcaster reported. The 149 passengers and eight crew were of 33 nationalities, the country's state broadcaster EBC reported on Sunday.
Ethiopia's prime minister offered condolences to victims' families.
"We hereby confirm that our scheduled flight ET 302 from Addis Ababa to Nairobi was involved in an accident today," the airline said in a statement on Sunday.
"It is believed that there were 149 passengers and eight crew on board the flight but we are currently confirming the details of the passenger manifest for the flight," according to the statement.
Earlier on Sunday, the airline said "search and rescue operations are in progress and we have no confirmed information about survivors or any possible casualties".
The spokesperson for the airline has confirmed to Reuters that 149 passengers and eight crew members were on board the plane at the time of the crash.
"Boeing 737-800MAX took off at 8.38 am local time from Addis Ababa and lost contact at 8.44 am. Search and rescue operation is in progress. It is believed that there were 149 passengers and 8 crew onboard the flight but we are currently confirming the details," a spokesperson said.
The Boeing 737-800MAX is the same type of plane as the Indonesian Lion Air jet that crashed last October, 13 minutes after takeoff from Jakarta, killing all 189 people on board.
The MAX is in service all around the world. Airlines such as the Ethiopian Airlines are using this aircraft, as it is the latest, the most fuel-efficient, short-range Boeing aircraft on the market.
Boeing said on its Twitter account that it was aware of the reports about Sunday's accident and was "closely monitoring the situation".


text:
We use cookies to help you get the best experience when using our site.
By continuing to use this website, you are agreeing to our cookies policy.
By continuing to use this website, you are agreeing to our cookies policy.
Salary band: £60k - £70k
Here is an excellent opportunity to work within a Top 200 firm working alongside Legal 500 chambers ranked and leading individuals within a reputable commercial department. If you are commercial contracts solicitor looking to flourish within a firm, known for the highest quality work, then this is a stand out opportunity. You will work upon large IP and IT contracts with some of the most recog...
Other Solicitor Jobs in Dartford
Salary band: £20k - £25k
Dartford based law firm are seeking a legal secretary to work in the family law department on a permanent basis. Previous family law experience welcomed but not essential as training can be given. However, candidates must have good audio and copy typing skills. Full or part time candidate applications considered. Salary negotiable depending on prior experience. Apply with CV for vacancy 19519.
Other Admin Jobs in Dartford
Salary band: £40k - £50k
Private Client Practitioner REF: YK1612 Salary: £40,000 to £45,000 dependent on experience Location: Dartford Job Type: Permanent, Full Time Job Section: Private Practice Experience: 2+ years experience Contact: Yoned Khan Company Benefits: Onsite parking, Bonus Structure Progressive Law Firm seeks experienced Private Client Practitioner. Managing and developing a caseload of mainly Probate fil...


text:
Ajax Login TitleClose
Returning Customer
I am a returning customer
Ajax Login TitleClose
Your Account Has Been Created!
Congratulations! Your new account has been successfully created!
You can now take advantage of member privileges to enhance your online shopping experience with us.
If you have ANY questions about the operation of this online shop, please e-mail the store owner.
A confirmation has been sent to the provided e-mail address. If you have not received it within the hour, please contact us.
Account Logout
You have been logged off your account. It is now safe to leave the computer.
Your shopping cart has been saved, the items inside it will be restored whenever you log back into your account.
Suitable for VegetariansStore in a cool dry, odour free place.Alpen8 - 12 Days Delivery*Please not Best Before/Expiration UK is DD/MM/YYYY
Start Positive with AlpenEnjoy a bright start to the day with our deliciously tasty, nutritionally balanced muesli. We've blended toasted wholegrain wheat flakes, creamy rolled oats, crunchy hazelnuts, almonds and juicy raisins to create this irresistible recipe that is packed full of Alpen positivity for a great day ahead!**Reducing consumption of sodium helps maintain normal blood pressure as part of a healthy diet & lifestyle. Sodium 0.10g per 100gEach spoonful contains...Signature toasted wholegrain wheat flakesCreamy rolled oatsCrunchy hazelnuts and almondsJuicy raisins Fancy a tasty breakfast that's full of good stuffWake up to Alpen... satisfaction by the spoonful!Every bowl is cram packed with an abundance of delicious ingredients.Alpen... breakfast at its peak! 100% natural ingredients, Wholegrain, High in fibre, Low in sat. fat, Suitable for vegetarians Check out alpenmornings to hear more about Alpen positivity and to get hints & tips from our health & wellbeing experts
Alpen Original Swiss Recipe Muesli 1.3kg JQTHDEWJE


text:
Filed under: ffmpeg ,a kin known as quest ,jessy lanza ,kaytranada ,lists ,songs of the 12 months category:better of ,classics ,featured ,mixes ,mp3 ,information
How to change a windows media audio pillar a mp3?
You have to bolt your itunes beforehand earlier than you may obtain something in the web. when you do not like to download from itunes which means paying, you should utilize the web to obtain music sort mp3 then just export it in itunes and you can transfer the music to your ipod. thoughts you that downloading music from the web is prohibited appropriately it is higher to purchase online if you want to support the artist.
audacity relating to relating to Uspertaining to the scenerya propos the AuthorBooks by means of Jon Kabat-ZinnBill Moyers ProgramVideos of Jon TeachingCustomer CommentsMindfulness Books in other Languages2zero16 CalendarCDs MP3s Wholesale FAQ MP3 FAQ CartHome regarding- relating to Us- with regard to the resolved- the Author- Books by Jon Kabat-Zinn- Bill Moyers Program- Videos of Jon Teaching- Customer Comments- Mindfulness Books in other Languages- 2016 Calendar CDs MP3s Wholesale FAQ MP3 FAQ Cart forty4 Not FoundYour cart (zero)
Latest DJ Remix Mp3 Songs
Page 1, showing1 - 2four of seven5 iPod and MP3 gamers earlier Page123foursubsequent Page
Note with regard to "Mp3gain professional"The creator ofMP3Doctorrecently renamed his "SuperMp3Normalizer" program to " Mp3gain professional ". i did not write down this new professionalgram, thus please don't e-mail me any support questions on it.should you're interested, here are the principle ceremonial variations between "Mp3acquire professional" and my, uh, "classic"(?) MP3acquire: "Mp3gain professional" does quantity normalizationinsidethe mp3, not simply between mp3s. fittingly if you happen to really feel a track is just too exhausted firstly (or center, or finish), then it may boost the amount only for that half. pretty serene, if that is what you need.The changes "Mp3achieve professional" makes arenotundo-ready. so as to make its fine-tuned advertjustments, it should re-encode the mp3 row., test it out should you're interested. however do not ask me any questions ;)
Best Youtube to MP3 Downloader and Converter
Filed under: ffmpeg ,a kin known as quest ,jessy lanza ,kaytranada ,lists ,songs of the 12 months category:better of ,classics ,featured ,mixes ,mp3 ,information