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Wax Seal Stamp Rose Square
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Seal only or Gift Box Set with Wax
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Gift Set Seal and 1 Gold Wax stick [Add $3.00]
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Brass Wax Seal with attached handle and Rose engraving to make a detailed impression.
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Imported from Florence and made by Italian craftsmen, this seal is 2" tall with a Fleur de Lys Handle and a square 3/4" impression.
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Wax Seal Stamp Rose Square
August 28, 2013
Reviewer: Jennifer L Joy from Portland, OR United States
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Thirsty Owl Riesling
Thirsty Owl
- New York
- United States
- Finger Lakes
Riesling grapes are very rarely blended with others in the development of wines, and for good reason. These pale grapes which originated in the cool Rhine Valley of Germany are notable for their 'transparency' of flavor, which allows the characteristics of their terroir to shine through in wonderful ways. The result of this is a wine which carries a wide range of interesting flavors quite unlike those found in other white wines, finished off with the distinctively floral perfume Riesling supplies so well. Many wineries in Germany and elsewhere tend to harvest their Riesling grapes very late often as late as January in order to make the most of their natural sweetness. Other methods, such as encouraging the noble rot fungus, help the Riesling grape varietal present some truly unique and exciting flavors in the glass, and the variety of wines this varietal can produce mean it is one of the finest and most interesting available anywhere.
To get some idea of how well established many of the wineries in New York state are, one only has to look to certain locations around the Hudson River area of the region, where there are several wineries which have vineyards dating back over three hundred years. This relatively long tradition of wine-making for a New World country has resulted in some truly exceptional wines, making the most of the imported grape varietals which thrive in the cooler, damper climatic conditions which characterize the state. New York today produces a wide range of wine styles, but is mostly known for its white wines made from such esteemed varietals as Riesling and Pinot Noir, and a few superb sparkling wines which demonstrate the strengths of the Chardonnay grapes grown there.
Of all the New World wine countries, perhaps the one which has demonstrated the most flair for producing high quality wines - using a combination of traditional and forward-thinking contemporary methods - has been the United States of America. For the past couple of centuries, the United States has set about transforming much of its suitable land into vast vineyards, capable of supporting a wide variety of world-class grape varietals which thrive on both the Atlantic and the Pacific coastlines. Of course, we immediately think of sun-drenched California in regards to American wines, with its enormous vineyards responsible for the New World's finest examples of Cabernet Sauvignon and Merlot based wines, but many other states have taken to viticulture in a big way, with impressive results. Oregon, Washington State and New York have all developed sophisticated and technologically advanced wine cultures of their own, and the output of U.S wineries is increasing each year as more and more people are converted to their produce.

subset: commoncrawl


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Skydiver Smoke
$28.00 As low as: $25.20The Skydiver Smoke canister is designed for the demanding needs of skydivers. Double sleeved and with a recessed smoke ejection port, the Skydiver can be attached directly to your boot, or with a mount. A tremendous 30,000 cubic feet of smoke unleashed in only 60 seconds is sure to leave a great smoke trail and enhance any demonstration jump.
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Business and Industry Petitions - Page 762
Online petitions related to business and industry, hosted at iPetitions, the leading site for online activism and grassroots democracy. Online petitions in this category include petitions on corporate activity, petitions demanding greater business accountability, and petitions drawing attention to industrial or corporate wrongdoing. Use an online petition to hold a company accountable and to change corporate behavior.
Create your own petition- Donkey Kong Team Alliance
Donkey Kong Team Alliance is a Nicktoon I was planning to make. I want to create an animated series ... - FIRE AT WILL
In the state of Maryland there's a policy set in play called "fire at will". The the average ameri... - Support The Brick Yard Outdoor Seating!
Dear Friends,Please help support The Brick Yard Restaurant & Bar on Union Street and its applica... - buis
laooz - 東大街的啓示 - 給監督的信
給 監 督 的 信 致 : 呂立有 (政府律師), 香港金鐘道66號, 金鐘道政府合署高座2樓, 律政司 及 建築事務監督 陳小雲 (屋宇署) 及 屋宇署署長 ... - 給監督的信 - 東大街的啟示 - 建築物條例第24(1)及14條的魔鬼
給 監 督 的 信 致 : 呂立有 (政府律師), 香港金鐘道66號, 金鐘道政府合署高座2樓, 律政司 及 建築事務監督 陳小雲 (屋宇署) 及 屋宇署署長 ... - Stop The Destruction Of Maximum Ride!
Maximum Ride has been a loved series for the past years. However, a group of loyal FanFictioners and... - Save TAB (Torah Academy of Brooklyn)
Save Our School I am an alumnus of Torah Academy High School of Brooklyn and a member of the TAB Alu... - Move the Eastern APA to the first week of January
For decades, philosophers have been trekking to various east coast US cities on Dec.27 for the Easte... - TEAM NATICAKES: Bring naticakes to Palomar Centre!
In a world not so far away, a little girl named Nati giggles with delight as she anticipat... - TEAM NATICAKES: Bring naticakes to Brighton Place Shop!
In a world not so far away, a little girl named Nati giggles with delight as she anticipat... - This Is Our Voice!
To Senate President Michael Waddoups, House Speaker Becky Lockhart and the Utah Legislature: We, ... - Caesary Official Mirror (Kong)
Sign if you wish to have an official mirror or copy of the official Caesary game, on Kongregate.com ... - Entertainment and Media
Hey my name is Ben Warren and I am awsome - Bring Back the Bath & Body Works Temptations Line of Fragrances.
The Temptations fragrances made by Bath & Body Works have been discontinued, much to the disma...


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Most Recent Articles In Fashion Scoops
Latest Fashion Scoops Articles
- Katy Perry Launches NFL Women's Apparel Sweater
- Katy Perry's RVN Revolution
- Phoebe English to Launch E-Commerce
More Articles By
RICH PICKINGS: Azerbaijan just gained a foothold on Paris' Place Vendôme. The State Oil Fund of the Republic of Azerbaijan (SOFAZ) has bought the building housing the Dior and Mikimoto fine jewelry boutiques, in addition to Valentino's headquarters, for 135 million euros, or $178.4 million, according to real estate adviser Cushman & Wakefield. The building was sold by real estate manager Axa Real Estate on behalf of one of its clients, it added, without providing further details. Cushman & Wakefield's Capital Markets Group advised SOFAZ on the acquisition, its first in France. Established by the father of Azerbaijani president Ilham Aliyev, SOFAZ is charged with collecting and managing Azerbaijan's revenues from the implementation of oil and gas agreements. Officials at Dior and Valentino were not immediately available for comment.


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You must earn 10,000 points to post other members' whiteboard
I will miss you buddy! My deepest sympathies to your family. RIP Dave
I posted this on the Gasbuddies Group's Facebook page: ~ EXPAPERMAKER, Dave, developed swolen ankles on Fri. the 23rd of January. He didn't want to go to the doctor, but his wife took him to the emergency room that evening. He was diagnosed with congestive heart failure, COPD and pneumonia. Saturday night, he died in his sleep. Dave's wife will not be keeping the site active but will read messages left there.
God bless you and your love ones.
RIP Expapmaker, your job here is done and it is time for you to move on. God's Blessings. Ed.
RIP David
Blessings through the tears for those who loved Dave...rejoice in KNOWING he has reached the Promised Shore that beckons us all, IF we receive God's Blessed Salvation!
Good bye to a great buddy! Dave Rest in Peace! You're in a better place now, no more pain and suffering! My condolences along with our thoughts and prayers to your family! Ron
RIP Dave
My condolences fro the family, Dave is no longer in pain , He was one of my first buddies and sure will be missed, Rest in Peace my buddy


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WMC - Larry ElderDavid Horowitz Freedom CenterWednesday, January 30, 2013 from 11:00 AM to 1:25 PM (PST) |
Who's Going
Loading your connections...
3 of your friends are going.
Share it to get more people on board! Share
Sara Haughty
San Francisco, California
San Francisco, California
Luke Picard
London, United Kingdom
London, United Kingdom
Rick Vespucio
Florence, Italy
Florence, Italy
When & Where
300 S. Doheny Dr.
Los Angeles,
Wednesday, January 30, 2013 from 11:00 AM to 1:25 PM (PST)
Add to my calendar


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Ms. Joanna C. Anders
Family Medicine, Davies Clinic PC
- 345 N Grant St
Canby, OR 97013 - 1 Office Location
- (503) 266-2066
Ms. Anders works in Canby, OR and specializes in Family Medicine. Ms. Anders is affiliated with Providence Willamette Falls Medical Center. Ms. Anders speaks English and Spanish.
Gender: Not Available
Years in Practice: Not Available
Languages Spoken: English, Spanish
Medical School: Not Available
Graduation Date: Not Available
Hospital Affiliations: Providence Willamette Falls Medical Center
Specialties: Family Medicine


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Science China Chemistry
Science China Chemistry is an academic journal co-sponsored by the Chinese Academy of Sciences and the National Natural Science Foundation of China, and published by Science China Press. The journal publishes high-quality, original results in both basic and applied research.
Categories of articles include:
Reviews that summarize representative results and achievements in a particular topic or an area, comment on the current situation, and advise on research directions
Research papers reporting on important original results in all areas of chemistry
Brief reports presenting short, timely reports of the latest important results
- 19 Volumes
- 154 Issues
- 2,951 Articles
- 1997 - 2015 Available between
Latest Articles
N-phosphoryl amino acid models for P-N bonds in prebiotic chemical evolution
Feng Ni, Chuan Fu, Xiang Gao, Yan Liu, Pengxiang Xu, Liu Liu… (January 2015)
Yufen Zhao (January 2015)
Defect density engineering for better graphene performance
Shengli Chen (January 2015)
- Journal Title
- Science China Chemistry
- Coverage
- Volume 40 / 1997 - Volume 58 / 2015
- Print ISSN
- 1674-7291
- Online ISSN
- 1869-1870
- Publisher
- Science China Press
- Additional Links
- Topics
- Industry Sectors
Previous Title | Print ISSN | Online ISSN |
Science in China Series B: Chemistry | 1006-9291 | 1862-2771 |
Continue reading...
To view the rest of this content please follow the download PDF link above.


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- Episode Description
Jim tries to blackmail Dana when he finds out she's having erotic dreams about him.
- Cast & Crew
Billi Bruno
Courtney Thorne-Smith
James Belushi
Larry Joe Campbell
Taylor Atelian
- Fan Reviews (0)
- Be the first to write a review!
- Trivia & Quotes
Quotes (2)
Dana: (sleep talking) All right, come on, come on, take me, let's go to the Gazebo. Oh, yeah, the Gazebo. Yeah, come on, take me, Jim. Oh, Jim, you naughty boy.
Dana: What are you doing? Jim: Good morning, Dana. Sleep well? Dana: Why aren't you working on the nursery? Jim: I'll get to that. You know what? I was thinking about something that would be perfect for your home, an new addition. Dana: Your grave? Jim: I love the way we banter. No, I was thinking of something a little more romantic, like a Gazebo. Dana: Gazebo? Jim: Yeah, a Gazebo. Come on. Haven't you ever dreamt of a romantic Gazebo. Dana: Gazebo? Gazebo! Oh. Jim: Dana, why, you look as you just saw a ghost, a very sexy ghost. Dana: I'm fine, just leave me alone, go saw something. Jim: I'm sorry. Have I been a naughty boy? Dana: Oh! You know! Jim: Know what, Dana? Know what, That you had a dirty dream about me? Dana: Oh god! Oh god! Jim: I think it was more like "Oh, Jim, oh, Jim!" But I answer to both! Dana: Jim, you can not tell anyone about this. Jim: Hmm, what's in it for me? Dana: Nothing. Jim: Oh, okay. Citizens, Dana's got the hots for her brother-in-law! Dana: Hey! God! Fine! At do you what? Jim: I'll tell you what I want, I want no more complaining, I want no more changes! Dana: Fine, fine, whatever, okay! Jim: All right. Your secret remains with me.
Notes (2)
ABC repeated this episode on February 28, 2006 & August 22, 2006.
Ratings: This episode was watched by 8.00 million viewers.
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1st case of a child coming down with GB Syndrome after H1N1 shot
Comments (1)
Posted 11/13/2009
"Arlene Connin said she was eager to protect Jordan and his brother, Lleyton, 7, against the flu. When she took the boys to their pediatrician for seasonal flu shots on Nov. 5, the provider said H1N1 vaccine was available, too."
Just curious.... do we know if they ALSO got the seasonal flu shot? The article is not clear on this aspect. It doesn't say that they only got the H1N1 unless I read this wrong.
Mom to Maya & Noah
What's on your mind?
Create a new postYour growing child
Your child: 9 years and up
BabyCenter focuses on the development of kids age 8 and younger. But we also have lots of great articles, tools, and other resources to help you raise your older child. Read More


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Credits & Info
- Uploaded
- May 4, 2012 | 9:39 AM EDT
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Author Comments
WASD or Arrows to move
J or CLICK to attack
N and M to change from range to melee weapon
Rated 5 / 5 stars
lol its funny cuz they are litte bitty guys like the snalis and i like it! (5 of 5 starts)
Rated 3 / 5 stars
good overall game
3 out of 5 stars
for whatever reasson i think theres a glitch i got 2 helmets and started switching them,as i switched it lowered my health,(i wasn't taking any enemy damage) and i had lowered my health to nothing by the time i stopped...
Rated 0.5 / 5 stars
0.5 Star Cant give more or less the game work but it has no sort of balance cant get past the first lvl without using more then 1 nuke also the looting are terrible Never found a new bow only 2 kind of helmet that i found and 3 weapons well to make sort make sure the game have everything to work and entertain everyone and placing a good balance first its easy a handful of villager then they nuke once i get 10 of them and once its start to rain the monster its over by the way the jump is over power taking no damage when landing so this is my review sorry for taking time to make it


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We've all seen ads for diet programs that make losing lots of weight seem as quick and easy as posing for the "before and after" photos. But as anyone who has tried to lose weight knows, it can be a struggle.
It's easy to feel frustrated when we set a weight-loss goal and then don't drop all the pounds we planned to lose. That's why the best way to lose weight — and keep it off — is not to go on a diet. (Yes, you read that right!) Instead, make being healthy your ultimate goal, not a specific number on the scale.
The ideal approach to losing weight is to focus on making small, specific changes that are easy to stick with in the long run — like not eating while watching TV, replacing a lunchtime soda with water, or walking to the bus instead of getting a ride.
So instead of going on an all-out diet or exercising until your knees shake, try making one small change. Use our personal plan to guide you.
When you're finished, you can print a goal sheet to inspire you as you work to turn your change into something that feels like a natural part of your life. This personal plan tool also offers examples of how other people started their own small-change transformations.
When you've worked on your first change long enough that it has become a habit, make another change. Then another. These small changes all add up to a more successful way to lose weight.
Note: All information is for educational purposes only. For specific medical advice, diagnoses and treatment, consult your doctor.
© 1995–2014 The Nemours Foundation/KidsHealth. All rights reserved.
Find out by selecting your child's symptom or health condition in the list below:
In This Issue
Download Winter 2015 (PDF)
Miracle Season, hosted by Dan Lewis and Molly Shen, aired Dec. 7, 2014 on KOMO 4 TV. The annual holiday special celebrates the remarkable lives of Seattle Children's patients. To donate now, visit https://giveto.seattlechildrens.org
Play Video
This 30-second video features Dr. Samuel R. Browd, attending neurosurgeon and medical director of Seattle Children's Sports Concussion Program, defining concussion, listing the signs and urging you to get medical care right away if you think your child may have a concussion.
In early 2014, Microsoft reached an incredible giving milestone with Seattle Children's – $5 million donated! Microsoft supports our efforts in so many ways: matching employee gifts, corporate grants, in-kind contributions and more. Thanks to the generosity of this amazing company, headquartered in our community, the Science Adventure Lab has the...


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Jolie is certainly a beautiful kitty and now she's looking for her forever home! This 8-month-old beauty is also quite the rambunctious little lady who loves playtime. All she needs is her favorite toy and someone to love her. Could that be you? Jolie is spayed, micro chipped and up-to-date on vaccinations. Visit her today at the Kentucky Humane Society's Main Campus or call 502.366.3355 for more information. View all adoptable pets at www.kyhumane.org.
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If the game freezes, just click inside it (games can get unresponsive if you click elsewhere on the page).
Yes |
No |
Shoot the incoming minions of hell. Shoot them in the head to save bullets. Don't let them chop you!
DanyshaMcshay20 16 days ago
Cant get pass level 7
astroid 3 months ago
it is scary But still i like it.
doomedspirit 4 months ago
lol this game
Natsumi 4 months ago
Like! ^_^
abdullah 6 months ago
hell is full so iam back
maizee 7 months ago
run sombodey
maizee 7 months ago
WILLY 8 months ago
what shall i do to deaft them
WILLY 8 months ago
WILLY 8 months ago
gamer1234 8 months ago
i love this game so much <3
vinz34 10 months ago
so hard
sift_heads about 1 year ago
i really like this game
IndraProBoyz about 1 year ago
WTF? is so hard
IndraProBoyz about 1 year ago
I won before to level 10
mys about 1 year ago
i like to kill someone
haji about 1 year ago
...tsoii pa gwapo pa ako na 2.......
chanchan about 1 year ago
level 8 is so very hard
Amin about 1 year ago
entenit lembab ...!!! aduhh.. apew cellop
Amin about 1 year ago
roy x3 hati kau keras...!!
NurJoker5 about 1 year ago
is a really nice game to play :) BUT SOMETIMES SO CREEPY !
shimi about 1 year ago
hindi ko po maintindihan
xxxxROOM1xxxx about 1 year ago
leoleo1020013 about 1 year ago
scary but not that scary
William about 1 year ago
this game is fanyastic
This game is part of a series: 13 Days in Hell
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Copy this code to your page |
Advertise here: Trafficflux


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The Wii U GamePad gives players all sorts of options and features when it comes to gaming. And with Miiverse, the built-in social network, players can share not only messages, but drawings and hand-written notes. We've collected some impressive Miiverse drawings made by various Wii U users out there. Check out the full gallery below, and share some of your own favorites in the comments section. Enjoy!
Continue reading:
- Pokken Tournament has a water stage
- Apex 2015 halted due to roof collapse
- Frogmind's Badland is headed to Wii U
- Sega is restructuring its business away from physical games
TAGS: miiverse


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Michael Andrew Bodree Sr, 62, of Robertsdale, AL, passed away on October 13, 2013, in the care of Covenant Hospice at West Florida Hospital in Pensacola, FL.
Michael is survived by his parents Mr. and Mrs. Bernard Bodree Jr, his sister Gay (Tony) Brewton, his children Rocky (Lindsay), Ben (Tiffany), Kit (Blakleigh) and Dakota (Kayla), five grandchildren, two nieces Brittney (David) Cobb, and Summer Brewton, one nephew Austin Brewton, and other family and friends.
The funeral service will be held on October 18, 2013 at First Baptist Church of Pensacola. The funeral will be held at noon with visitation beginning at ten.
Funeral arrangements are being handled by Pensacola Memorial Gardens.
In lieu of flowers memorial donations may be given to your charity of choice. The family of Michael Andrew Bodree wishes to thank Doctor Maged N. Nashed, the nursing staff at West Florida Hospital, the staff at Life Care center of Pensacola, and the angels at Covenant Hospice.


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Ask Fmany of the chickies F7in my B♭mpen They'll E♭7tell you I'm the biggest mother A♭hen I C♯love them all and all of them love B♭mme G Because the Fmsystem works the Fdimsystem called C7reciFdimprociCty C7 FmStandard guitar tuning:EADGBENo capoC7 G
C7 Fm
FGot a little motto FmAlways sees me through C7When you're good to Mama Mama's good to you FmC7 G
FThere's a lot of favors FmI'm prepared to do C7You do one for Mama She'll do one for youFmF G
FThey B♭say that life is B♭m6tit for tat And Fthat's the way I live So DmI deserve a Flotta tat For G7what I've got to give C7 FDon't you know that this hand FmWashes that one too C7When you're good to Mama C♯7Mama's C7(♯5)good to youFm C7C7 Fm
FIf you want my gravy FmPepper my ragout C7Spice it up for Mama She'll get hot for youFmC7
FWhen the pass that basket FmFolks contribute to C7You put in for Mama She'll put out for youFmF G
FThe B♭folks atop the B♭m6ladder Are the Fones the world adores So Dmboost me up my Fladder kid And G7I'll boost you up yoursC7 FLet's all stroke together FmLike the Princeton Crew C7When you're strokin' Mama C♯9Mama's C9strokin' yoF7u So B♭what's the one conG7/Bclusion I can Fbring this numbeD7r to? Gm7When you're good to G7Mama C♯7Mama's C7(♯5)good to FmyouGm7 C7Fm
Gm7 C7 C7(♯5)
Terms and Privacy | API | Just Added Tabs | Tags | How to Read Guitar Tab | Music Theory | Contact/Support
© 2015, Songsterr.com.
© 2015, Songsterr.com.


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- melix is available for meet-ups
- female from Cyprus
- Photographer from Cyprus, Larnaca. I speak Dutch, Greek and English. I love painting and reading books. My blog: http://laughingtriangles.tumblr.com/ Facebook: http://www.facebook.com/groups/melixph…
- flowerpunk
- female () from Nicosia, Cyprus
- Hey. I'm new here, and I already love it
- magissoulla33
- female (25) from Cyprus


text:
Winnipeg Free Press - PRINT EDITION
Better access brings relief
Pain sufferer sparks Handi-Transit review
When Diane Driedger beat breast cancer in 2009, she also lost her Handi-Transit service, even though chronic fibromyalgia often still laid her low with pain and fatigue.
At the time, the former head of the Manitoba League of Persons with Disabilities was working on her PhD at the University of Manitoba. When she had a flare-up, getting to campus by regular transit, especially in the winter, often left her so drained she had trouble concentrating in class.
That prompted Driedger to complain to the Manitoba Human Rights Commission, alleging Handi-Transit did not adequately accommodate people with episodic conditions -- illnesses such as lupus, multiple sclerosis or arthritis that flare up badly one day, but can be tolerable the next. People like Driedger often failed to meet the key test for Handi service -- they were able to walk 175 metres outside, but not all of the time and not without a disabling level of pain and fatigue.
On Monday, after four years of negotiations, the commission announced it had brokered a deal between Driedger and the city that will expand Handi-Transit's assessment process to include episodic pain and fatigue as barriers to mobility. Later this fall, when Handi-Transit rewrites its assessment manual, it will better reflect the effects of illnesses with occasional flare-ups, which could allow more people with chronic illnesses to access the specialized service.
-- Tanya Misseghers, of the Manitoba office of the Arthritis Society
It's not clear how many more people may now apply for or receive Handi-Transit approval, or how the service's budget may be affected. Driedger says she knows of several people with chronic illnesses who, like her, were denied Handi-Transit despite confirmation from a doctor.
But city spokeswoman Michelle Bailey said the city is not bracing for a wave of new applicants.
"We do not anticipate an increase in registrations for Handi-Transit services, as we have always considered and enrolled applicants with episodic disabilities including, but not limited to, individuals with arthritis, multiple sclerosis, lupus, cancer, diabetes and fibromyalgia," she said in an emailed statement.
Driedger jokes that because she's a "sturdy Mennonite," people often don't realize she suffers from a chronic disability because her pain and fatigue are largely invisible.
Asked how the city will ensure people using Handi-Transit are legitimately having a flare-up, Driedger said the honour system should work.
"I think we have to trust people here to define their own situation," she said.
Typically, Handi-Transit is not a convenient way to get around. Many trips must be booked in advance, limiting flexibility, and wait times can be frustrating. And, many people with episodic illnesses can cope with regular transit, have their own car or can arrange rides.
"Any step forward to increasing accessibility of services for people with inflammatory types of arthritis is very important to us and we consider that very good news," said Tanya Misseghers, the community engagement and marketing co-ordinator for the Manitoba office of the Arthritis Society.
Of the 250,000 Manitobans with arthritis, about 20 per cent suffer from the inflammatory type that can be episodic and debilitating but often invisible.
Disability advocates have long criticized the city's rigid Handi-Transit eligibility rules. The latest deal brokered by the Manitoba Human Rights Commission is at least the third time the commission has intervened on behalf of people with disabilities in disputes with the city.
Earlier this year, a 71-year-old woman with intellectual and physical disabilities won a $2,000 settlement after the commission ruled she had been unfairly denied Handi-Transit services that helped her get to work.
Following a complaint several years ago, the commission ordered Handi-Transit must serve people with Alzheimer's disease and other forms of dementia. In that case, negotiations between the commission the city and the complainant also took four years.
Republished from the Winnipeg Free Press print edition August 26, 2014 0
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6 thoughts on "Dark Horse Comics Is Brought to Google Play"
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Influence of age and 17beta-estradiol on kisspeptin, neurokinin B, and prodynorphin gene expression in the arcuate-median eminence of female rhesus macaques.
ABSTRACT The neuropeptides kisspeptin, neurokinin B, and dynorphin A (collectively abbreviated as KNDy) are, respectively, encoded by KiSS-1, NKB, and PDYN and are coexpressed by neurons of the hypothalamic arcuate nucleus (ARC). Here, using quantitative real-time PCR, we examined age-related changes in the expression of genes encoding KNDy and associated receptors G protein-coupled receptor 54 (encoded by GPR54), neurokinin 3 receptor (encoded by NK3), and kappa-opioid receptor (encoded by KOR), in the female rhesus macaque ARC-median eminence (ARC-ME). Expression of KiSS-1 and NKB was highly elevated in old perimenopausal compared with young or middle-aged premenopausal animals. To test whether these age-related changes could be attributed to perimenopausal loss of sex steroids, we then examined KNDy, GPR54, NK3, and KOR expression changes in response to ovariectomy (OVX) and exposure to 17beta-estradiol (E(2)). Short-term (7 months) OVX (with or without 1 month of estrogen replacement) failed to modulate the expression of any of the KNDy-related genes. In contrast, long-term ( approximately 4 yr) OVX significantly increased KiSS-1 and NKB expression, and this was reversed by E(2) administration. Finally, we examined the expression of KNDy-related genes in young adult females during the early follicular, late follicular, or midluteal phases of their menstrual cycle but found no difference. Together, the results suggest that short-term alterations in circulating E(2) levels, such as those occurring during the menstrual cycle, may have little effect on the ARC-ME expression of KNDy and associated receptors. Nevertheless, they clearly demonstrate that loss of ovarian steroid negative feedback that occurs during perimenopause plays a major role in modulating the activity of KNDy circuits of the aging primate ARC-ME.
- [Show abstract] [Hide abstract]
ABSTRACT: Kisspeptin neurons in the rostral periventricular area of the third ventricle (RP3V) play a key role in relaying the positive feedback effects of estradiol that activate gonadotropin-releasing hormone (GnRH) neurons and drive a surge in the GnRH/luteinizing hormone (LH) level. However, the precise role of kisspeptin neurons during female reproductive senescence remains unclear. Focusing on middle-aged intact female mice with irregular estrous cycles, we found a parallel decline in c-Fos-positive kisspeptin neurons and c-Fos-positive GnRH neurons at the time of the GnRH/LH surge. Furthermore, in kisspeptin neurons, the expression of estrogen receptor α (ERα), but not progesterone receptor (PR), decreased with age. Interestingly, some kisspeptin neurons in the RP3V, but none of the GnRH neurons in the rostral preoptic area (rPOA), had a characteristic cellular senescence in middle-aged mice and old mice. These data suggest that, among the groups of neurons involved in reproductive control, the kisspeptin neurons in the RP3V are likely among the earliest to undergo aging processes and thus participate in initiating the early reproductive decline.Neurobiology of aging 10/2013; · 5.94 Impact Factor - SourceAvailable from: Germano GuerraM Guida, F Zullo, B Buonomo, M L Marra, V Palatucci, R Pascale, F Visconti, G Guerra, Ml Spinelli, A Di Spiezio Sardo[Show abstract] [Hide abstract]
ABSTRACT: Menopause is characterized by depletion of ovarian follicles, a reduction of ovarian hormones to castrate levels and elevated levels of serum gonadotropins from the anterior pituitary gland. Although this process has significant repercussions throughout the body and affects a large proportion of our society, the neuroendocrine control mechanisms that accompany menopause are poorly understood. This review aims to examine rigorously the most accredited literature to provide an update about our current understanding of the role of the hypothalamic-pituitary axis in the onset of and transition into female reproductive senescence, focusing on the role of some specific neuropeptides in regulating the HPG axis and on their effects on several menopausal symptoms, especially referring to the cardiovascular risk, to open up new horizons for new therapeutic strategies.Translational medicine @ UniSa. 05/2012; 3:25-41. - [Show abstract] [Hide abstract]
ABSTRACT: In many species, sexual activity varies on a seasonal basis. Kisspeptin (Kp), a hypothalamic neuropeptide acting as a strong activator of GnRH neurones, plays a critical role in this adaptive process. Recent studies reported that two other neuropeptides, namely neurokinin B (NKB) and dynorphin (DYN), are co-expressed with Kp (and therefore termed KNDy neurones) in the arcuate nucleus and these peptides are also thought to influence GnRH secretion. The aim of this study was to establish whether hypothalamic NKB and DYN expression is photoperiod-dependent in a seasonal rodent, the Syrian hamster, which exhibits robust seasonal rhythms in reproductive activity. The majority of Kp neurones in the arcuate nucleus co-express NKB and DYN and expression of all three peptides is decreased in short, compared to long, photoperiod leading to a 60% decrease in the number of KNDy neurones in photo-inhibitory conditions. In seasonal rodents, RFRP neurones of the dorsomedial hypothalamus are also critical for seasonal reproduction. Interestingly, NKB and DYN are also expressed in the dorsomedial hypothalamus but do not co-localise with RFRP-immunoreactive neurones, and expression of both NKB and DYN is higher in short photoperiod, which is opposite to the short day inhibition of RFRP expression. In conclusion, this study shows that NKB and DYN display different photoperiodic variations in the Syrian hamster hypothalamus. In the arcuate nucleus, NKB and DYN, together with Kp are down-regulated in short photoperiod, whereas in the dorsomedial hypothalamus NKB and DYN are up-regulated in short photoperiod. This article is protected by copyright. All rights reserved.Journal of Neuroendocrinology 06/2014; · 3.51 Impact Factor
Page 1
Influence of Age and 17?-Estradiol on Kisspeptin,
Neurokinin B, and Prodynorphin Gene Expression in
the Arcuate-Median Eminence of Female Rhesus
Dominique H. Eghlidi, Gwendolen E. Haley, Nigel C. Noriega, Steven G. Kohama,
and Henryk F. Urbanski
Division of Neuroscience (D.H.E., G.E.H., S.G.K., H.F.U.), Oregon National Primate Research Center,
Beaverton, Oregon 97006; Department of Physiology and Behavior (N.C.N.), University of California
Davis, Davis, California 95618; and Departments of Physiology and Pharmacology (H.F.U.), and
Behavioral Neuroscience (G.E.H., H.F.U.), Oregon Health and Science University, Portland, Oregon 97239
The neuropeptides kisspeptin, neurokinin B, and dynorphin A (collectively abbreviated as KNDy)
are, respectively, encoded by KiSS-1, NKB, and PDYN and are coexpressed by neurons of the hy-
pothalamic arcuate nucleus (ARC). Here, using quantitative real-time PCR, we examined age-
related changes in the expression of genes encoding KNDy and associated receptors G protein-
coupled receptor 54 (encoded by GPR54), neurokinin 3 receptor (encoded by NK3), and ?-opioid
receptor (encoded by KOR), in the female rhesus macaque ARC-median eminence (ARC-ME). Ex-
pression of KiSS-1 and NKB was highly elevated in old perimenopausal compared with young or
middle-aged premenopausal animals. To test whether these age-related changes could be attrib-
uted to perimenopausal loss of sex steroids, we then examined KNDy, GPR54, NK3, and KOR
expression changes in response to ovariectomy (OVX) and exposure to 17?-estradiol (E2). Short-
term (7 months) OVX (with or without 1 month of estrogen replacement) failed to modulate the
expression of any of the KNDy-related genes. In contrast, long-term (?4 yr) OVX significantly
increased KiSS-1 and NKB expression, and this was reversed by E2administration. Finally, we ex-
amined the expression of KNDy-related genes in young adult females during the early follicular,
late follicular, or midluteal phases of their menstrual cycle but found no difference. Together, the
results suggest that short-term alterations in circulating E2levels, such as those occurring during
the menstrual cycle, may have little effect on the ARC-ME expression of KNDy and associated
receptors. Nevertheless, they clearly demonstrate that loss of ovarian steroid negative feedback
the aging primate ARC-ME. (Endocrinology 151: 3783–3794, 2010)
the hypothalamic-pituitary-gonadal (HPG) axis. In
contrast to rodents, the age-associated decline in circu-
lating sex-steroid hormone concentrations in primates
is thought to stem primarily from a depletion of ovar-
ian follicles, rather than from sex-steroid-independent
changes within the hypothalamus (1). Even so, hypotha-
lamic changes do occur in perimenopausal women and
enopause marks the end of reproductive function
for women and is characterized by a disruption of
may have a major impact on the activity of the HPG axis
as well as other physiological functions. Currently, there
is clear evidence that a population of cells located in the
arcuate nucleus (ARC) of the hypothalamus coexpress
kisspeptin, neurokinin B, and dynorphin A (2); here, we
refer to these neuropeptides collectively by the abbrevia-
tion KNDy (2). Importantly, these neurons play a major
role in modulating the release of GnRH from the hypo-
thalamus and also show marked changes in their activity
ISSN Print 0013-7227
Printed in U.S.A.
Copyright © 2010 by The Endocrine Society
doi: 10.1210/en.2010-0198 Received February 18, 2010. Accepted May 6, 2010.
First Published Online June 2, 2010
ISSN Online 1945-7170Abbreviations: ARC, Arcuate nucleus; AVPV, anteroventral periventricular nucleus; E2,
17?-estradiol; EF, early follicular; HPG, hypothalamic-pituitary-gonadal; KNDy, kisspeptin,
neurokinin B, and dynorphin A; LF, late follicular; MBH, medial basal hypothalamus; ME,
median eminence; ML, midluteal; P4, progesterone; OVX, ovariectomized.
Endocrinology, August 2010, 151(8):3783–3794endo.endojournals.org
Page 2
after menopause (1). Consequently, it is likely that ARC
KNDy neurons serve as conduits for estrogen's action on
GnRH neurons. Although GnRH neurons appear to ex-
press estrogen receptor ? (3) they do not express estrogen
receptor ? (ER?) (4–6), whereas kisspeptin neurons do
type that mediates estrogen negative feedback on GnRH
neuronal activity (16, 17).
Kisspeptin and neurokinin B appear to be necessary
for normal GnRH secretion and reproductive function,
because mutations in their receptor genes, G protein-
coupled receptor 54 (encoded by GPR54) (18, 19) and
neurokinin 3 receptor (encoded by NK3) (20), respec-
tively, result in hypogonadotropic hypogonadism.
Also, it has been shown in rodents, sheep, and nonhu-
man primates that a prepubertal rise in GPR54 and
KiSS-1 expression leads to the activation of GnRH,
which in turn stimulates the pubertal rise of LH release
from the anterior pituitary gland (21–25). Kisspeptin
and neurokinin B neurons of the ARC appear to exert
their influence directly on GnRH neurons via GPR54
and NK3 receptors, respectively, located in the median
eminence (ME) (26–28), thereby modulating LH re-
lease (29, 30). Dynorphin A (encoded by PDYN) is also
thought to play a role in mediating sex-steroid feedback
to GnRH neurons (31), by acting through the ?-opioid
receptor (encoded by KOR) to inhibit LH release
Although expression of KiSS-1, NKB, and PDYN (col-
lectively referred to here as KNDy) in the ARC clearly
plays an important role in controlling the HPG axis, its
precise function during primate aging is less clear. In ro-
hypothalamus precede the onset of reproductive senes-
cence (35, 36). For example, reduced estrogen-mediated
drive from kisspeptin neurons in the anteroventral
periventricular nucleus (AVPV) to GnRH neurons ap-
function (37). However, an AVPV kisspeptin neuronal
population is not conserved in the sheep, nonhuman pri-
medial basal hypothalamus (MBH) (38–40).
insights into the role of hypothalamic KNDy neuropep-
tides during aging. Using the female rhesus macaque as a
translational clinical model, we focused on two specific
questions. 1) How does the expression of KNDy and as-
sociated receptor-encoding genes change in the ARC-ME
during aging? 2) What role do sex steroids play in medi-
ating these age-related changes?
Materials and Methods
Adult female rhesus macaques (Macaca mulatta) were cared
for by the Division of Animal Resources at the Oregon National
Primate Research Center (ONPRC) in accordance with the Na-
tional Research Council's Guide for the Care and Use of Labo-
ratory Animals. The animals were housed indoors under con-
trolled conditions: 24 C temperature; 12-h light, 12-h dark
photoperiods (lights on at 0700 h); regular meals at 0800 and
1500 h (Purina High Protein Monkey Chow; Purina Mills, Inc.,
the following Institutional Animal Care and Use Committee-
from the ARC-ME and quantification of KiSS-1, NKB, PDYN,
GPR54, NK3, and KOR expression using real-time PCR.
Experiment 1: effect of age in ovary-intact animals
Postmortem rhesus macaque brain tissue was obtained
through the ONPRC Tissue Distribution Program. The ovary-
intact females had previously been involved in a cross-sectional
2.1 yr; n ? 5); middle-aged premenopause, regular cycling
(25.8 ? 0.4 yr; n ? 4). Serum, collected at time of necropsy, was
(P4). Mean E2concentrations were 40 ? 6, 71 ? 35, and 62 ?
16 pg/ml for the young, middle-aged, and old animals, respec-
low or undetectable (?0.2–0.8 ng/ml) P4concentrations, most
of the animals were assumed to be in the follicular phase of their
menstrual cycle. The one exception was a young animal that
showed elevated P4concentrations (2.4 ng/ml) and so was as-
sumed to be in the midluteal phase; all of the animals in the old
Experiment 2: effect of short-term E2treatment in
young ovariectomized (OVX) animals
Adult rhesus macaques were bilaterally OVX and approxi-
mately 7 months later were assigned to one of following exper-
imental groups: untreated controls (OVX; mean ? SEM age,
9.7 ? 0.3 yr; n ? 4) or treated with E2for 1 month (OVX?E2;
mean ? SEM age, 9.6 ? 0.8 yr; n ? 4). Terminal serum E2con-
gene expression levels of these two groups were analyzed to-
gether with those of the young gonad-intact animals from ex-
periment 1 (13.9 ? 2.1 yr, n ? 5 per group), which showed a
mean serum E2concentration of 40 ? 6 pg/ml at necropsy.
Experiment 3: effect of long-term E2treatment in
old OVX animals
Old female macaques were bilaterally OVX and approxi-
mately 2.5 months later were assigned to one of the following
experimentalgroups:untreatedcontrols(OVX;mean? SEMage,
mean ? SEM age, 27.0 ? 0.9 yr; n ? 4). Serum E2concentrations
concentrations were either undetectable or just above the detec-
Eghlidi et al.
E2Effects in the Aging PrimateEndocrinology, August 2010, 151(8):3783–3794
Page 3
to the undetectable levels, the mean concentration was 26 ? 2
pg/ml. In contrast, the mean E2concentration shown by the
OVX?E2group was 124 ? 11 pg/ml. For reference, the gene
expression levels of these two groups were analyzed together
experiment 1 (mean ? SEM age, 25.8 ? 0.4 yr; n ? 4), which
showed a mean serum E2concentration of 62 ? 16 pg/ml at
Experiment 4: effect of menstrual cycle phase
Adult ovary-intact macaques were monitored daily for men-
strual cycle regularity, and cycle phase was corroborated with
serum assays for E2and P4. Necropsies were performed at three
the start of the last menses); the animals were 10.5 ? 0.6 yr old
(n ? 4) and showed a characteristic rise in serum E2concentra-
tions (65 ? 19 pg/ml) and undetectable (?0.2 ng/ml) P4con-
centrations; 2) the late follicular phase (LF) (11.8 ? 1.3 d after
the start of the last menses); the animals were 12.2 ? 0.2 yr old
43 pg/ml) and the start of a P4rise (0.5 ? 0.1 ng/ml); and 3) the
midluteal phase (ML) (15.8 ? 1.7 d after the start of the last
characteristic low E2concentrations (39 ? 4 pg/ml) and high
P4concentrations (4.9 ? 0.8 ng/ml). Phases of the menstrual
cycle were further corroborated at necropsy by histological
examination of the endometrium: EF, thin endometrium and
ciliated oviduct; LF, thin, proliferative endometrium and ovi-
duct ciliated and fully secretory; and ML, thickened endome-
trium (41).
Experiment 5: 24-h changes in gene expression in
young OVX animals
To examine whether gene expression differs between the day
and night, we compared ARC-ME gene expression at 0100 h
the same 12-h light, 12-h dark photoperiods. During the night-
time tissue collections, the animals' eyes were covered (to shield
from light exposure).
In experiments 2 and 3, the E2was administered via sc
viously described (42). The resulting circulating hormone con-
centrations were similar to those typically observed during the
late follicular phase of the menstrual cycle. In experiment 3, the
implants were replaced after each year of treatment to ensure
sustained long-term delivery of the steroid.
Hormone assays
Blood samples were centrifuged and the supernatant serum
was stored at ?80 C until time of assay. Serum E2and P4con-
centrations were determined by the ONPRC Endocrine Tech-
nology and Support Core using a chemiluminescence-based au-
tomatic clinical platform (Immulite 2000; Siemens Healthcare
was 20 pg/ml and 0.2 ng/ml, respectively. The intra- and inter-
assay coefficients of variation were less than 15% for both
Tissue collection and RNA extraction
Postmortem brain tissue used in this study was obtained
through the ONPRC Tissue Distribution Program. The ani-
mals were sedated with ketamine (15–25 mg/kg im) followed
by pentobarbital sodium (25–30 mg/kg iv), a procedure con-
sistent with the recommendations of the American Veterinary
Medical Association's Panel on Euthanasia. Each brain was
flushed with 1 liter of 0.9% saline via a cardiac catheter, and
the MBH was removed and preserved in RNAlater (Ambion,
Austin, TX) for 2 wk. A coronal slice encompassing the
ARC-ME was removed and stored frozen at ?80 C. The
boundaries for this tissue block included the exterior ventral
edge of the ME, lateral cuts midway between the third ven-
tricle and the optic nerve, an anterior cut along the posterior
edge of the optic chiasma, a posterior cut just anterior to the
mammillary bodies, and a cut 1 mm dorsal to the base of the
third ventricle (i.e. based on stereotaxic coordinates, this rep-
resents the border between the ARC and the ventromedial
hypothalamus). Subsequently, each ARC-ME block was in-
dividually homogenized using a PowerGen rotor-stator ho-
mogenizer (Fisher Scientific, Pittsburgh, PA), and RNA was
extracted using a QIAGEN (Valencia, CA) RNeasy Mini Kit.
Integrity of the RNA was verified using spectrophotometry
(Agilent 2100 Bioanalyzer Nanodrop; Agilent Technologies,
Santa Clara, CA), and acceptable A260/A280ratios were used
for the determination of RNA concentrations. For each
sample, 1 ?g RNA was then converted to cDNA using
random hexamers and the SuperScript III First-Strand Syn-
thesis System for RT-PCR (Invitrogen Life Technologies,
Carlsbad, CA).
Primers and probes
Table 1 depicts the TaqMan primers and probes that were
used for detection of the neuropeptide genes (KiSS-1, NKB,
and KOR) as well as for two housekeeping genes (ALG9 and
the rhesus ARC-ME across various sex-steroid environments
(43). The nucleotide sequences for these genes either were ob-
tained from previously reported studies or were designed using
Primer Express 2.0 software [Applied Biosystems (ABI), Foster
City, CA] and were based on NCBI rhesus macaque reference
gen and Applied Biosystems (Carlsbad, CA), respectively. RT-
PCR, using ARC-ME and ovarian RNA samples (as positive
controls), was used to validate each of the primer sets by pro-
ducing amplicons of a predicted size.
TaqMan quantitative RT-PCR
Quantitative real-time PCR data were obtained using a
7900HT Fast Real-Time PCR thermal cycler and sequence de-
tection system software (version 2.2.1; ABI). Initially, pooled
cDNA was used to create standard curves for each gene, and the
experimental samples were subsequently diluted accordingly so
as to fall within the linear part of the curve. The PCR mixtures
contained 5 ?l TaqMan Universal PCR Master Mix, 0.3 ?l of
each specific forward and reverse primer (300 nM final concen-
tration), 0.25 ?l of specific probe (250 nM final concentration),
Endocrinology, August 2010, 151(8):3783–3794 endo.endojournals.org
Page 4
and 2 ?l of cDNA. The reaction sequence included 2 min at 50
C, 10 min at 95 C, and 50 cycles of 15 sec at 95 C and 1 min at
60 C. Automatic baseline and threshold levels were determined
by ABI sequence detection system software (version 2.2.1.), and
the final expression values are expressed as ratios relative to the
average of two reference genes, ALG9 and RPL13A. Individual
examined on separate plates. A negative control included the
omission of cDNA templates from the reaction mixture.
Statistical analysis
between group means. In experiment 5, differences between the
two groups were analyzed using the Mann Whitney U test. P ?
0.05 was considered to be statistically significant.
KiSS-1, NKB, and PDYN
(P ? 0.05) in old perimenopausal animals compared with
the young and middle-aged premenopausal animals (Fig.
1, A and B). PDYN expression levels did not differ be-
tween the experimental groups (Fig. 1C).
In the young animals (experiment 2), the expression
levels of KiSS-1, NKB, or PDYN were not significantly
gonadal status (ovary-intact vs. OVX) or short-term E2
replacement (OVX?E2) (Fig. 2, A–C). In contrast, in the
and NKB showed a significant (P ? 0.01) increase after
long-term OVX, and this was blocked by long-term E2
replacement (OVX?E2) (Fig. 2, D and E). As with the
of gonadal status or E2treatment on PDYN expression
(Fig. 2F).
No differences in KiSS-1, NKB, or PDYN expression
were observed across the menstrual cycle of young adult
animals (Fig. 3, A–C).
GPR54, NK3, and KOR
in the young, middle-aged premenopausal, and old peri-
menopausal ovary-intact animals (Fig. 4, A–C). Further-
more, there was no obvious effect of gonadal status or
these receptor-encoding genes (Fig. 5, A–F). Similarly, no
served across the menstrual cycle of young adult animals
(Fig. 6, A–C).
Gene expression at 0100 and 1300 h
We considered the possibility that our failure to detect
any significant changes in PDYN expression may have
TABLE 1. TaqMan real-time PCR primers and probes
Gene name
Primer sequence (5?–3?) Probe sequence (5?–3?)
accession ID
Reference sequences for each of the targeted Macaca mulatta genes can be accessed via GenBank accession ID.
Eghlidi et al.
E2Effects in the Aging PrimateEndocrinology, August 2010, 151(8):3783–3794
Page 5
stemmed from performing necropsies at a time of day
when transcription was at a diurnal low point (44, 45),
with less obvious between-group differences. As a result,
we compared the expression of PDYN as well as KiSS-1,
NKB, GPR54, NK3, and KOR at two different times of
day (Table 2). Although there was a tendency for some of
the genes to show diurnal differences in expression, only
GPR54 showed a statistically significant (P ? 0.05) dif-
1300 h than at 0100 h (i.e. they were higher at the ap-
proximate time of day when all of the necropsies in ex-
periments 1–4 were performed). Importantly, PDYN ex-
pression was similar regardless of whether the necropsies
were performed in the middle of the day or in the middle
of the night.
Like women, old female rhesus macaques undergo meno-
concentrations during the perimenopausal period (46).
which to examine the neuroendocrine mechanisms asso-
ARC increases significantly during the perimenopause.
This observation is consistent with previous findings that
hypothalamic KiSS-1 expression increases in postmeno-
pausal women and macaques (47, 48). However, in the
present study, we measured serum E2and P4concentra-
be sure whether the three different groups had sustained
periods of differential serum sex-steroid concentrations;
furthermore, based on individual animal menstruation
records and very low or undetectable P4concentrations,
be in the early follicular phase of the menstrual cycle or to
be acyclic and hence showed a less dramatic age-related
from an attenuation of cyclic ovarian steroid concentra-
tions. This possibility was examined in experiment 3,
which showed that long-term removal of E2, through
OVX, significantly enhances KiSS-1 and NKB gene ex-
pression in the ARC-ME. Overall, this finding is in agree-
in neuronal hypertrophy in the ARC (49–51). Impor-
tantly, our data also demonstrate that the post-OVX in-
blocked by treatment with E2. Furthermore, because the
was highly correlated with that of NKB (r2? 0.95), it is
likely that sex-steroid control of these two neuropeptide
systems is mediated through a common pathway. Collec-
tively, findings from the present and previously published
NKB expression stem directly from changes in the sex-
steroid environment rather than from aging per se. Con-
sequently, these gene expression changes are likely to be
FIG. 1. Age-related changes in KNDy expression in the ARC-ME of
adult female rhesus macaques, revealed by real-time PCR. The values
represent the means (?SEM) from young (13.9 ? 2.1 yr), middle-aged
(22.2 ? 0.4 yr), and old perimenopasual (25.8 ? 0.4 yr) animals. A
and B, Expression of KiSS-1 and NKB increased significantly in the old
animals relative to the other adults groups; C, PDYN showed no age-
related change in expression. *, P ? 0.05, ANOVA followed by
Student-Newman-Keuls test.
Endocrinology, August 2010, 151(8):3783–3794 endo.endojournals.org
Page 6
associated with a consequence rather than with a root
cause of nonhuman primate and human reproductive
In contrast to these KiSS-1 and NKB responses shown
after long-term OVX and E2replacement, the responses
observed in our short-term, 1-month es-
trogen treatment model (experiment 2)
were less obvious; although there was a
similar overall trend and some individu-
als clearly showed changes similar to
those of animals in experiment 3, the re-
plausible that larger group sizes would
have disclosed more clear-cut between-
group differences, but the present results
may also suggest that short-term manip-
ulation of the sex-steroid environment is
simply too subtle to significantly modu-
late KiSS-1 and NKB expression in the
ARC of young adults. This view is cor-
roborated by our observation that nei-
ther gene showed a significant change in
expression level when examined at three
mediately after the LH surge), and ML
phases. Because we failed to observe an
increase in ARC KiSS-1 expression dur-
conclude that kisspeptin plays no role in
the kisspeptin (i.e. protein) preovulatory
LH surge of primates. However, such a
conclusion may not be warranted, espe-
cially in view of the recent report of pre-
ovulatory elevated Kiss-1 expression in
experiment 4 were correctly assigned to
the three different phases of the men-
strual cycle, based on multiple criteria,
including days after menstruation, uter-
ine histology, and serum E2and P4con-
centrations, the serum E2concentrations
have been expected. This raises the pos-
sibility that some of the animals in this
group had already ovulated and were in
surge. Consequently, we cannot rule out
the possibility that in primates, a transi-
tory increase in KiSS-1 and/or NKB oc-
curs relatively early during the LF phase
of the menstrual cycle, before the pre-
ovulatory LH surge. Nevertheless, our
data emphasize that there is no obvious
sustained modulation of KiSS-1 or NKB expression across
the natural menstrual cycle, which is in contrast to the
marked changes observed during aging and after experi-
mental manipulation of the sex-steroid environment.
FIG. 2. Effect of short-term (A–C) or long-term (D–F) estrogen replacement on KNDy
expression in the ARC-ME of young adult and old female rhesus macaques. The values
represent the means (?SEM) from ovary-intact (black bars), OVX (white bars), and OVX?E2
(gray bars) animals. In the young age group (A–C), the expression level of KNDy was similar
regardless of treatment. In the old animals (D and E), the expression of KiSS-1 and NKB
was significantly elevated after OVX and restored to ovary-intact levels by the E2
treatment; the expression of PDYN (F) in the old animals was not different between
treatments. **, P ? 0.01, ANOVA followed by Student-Newman-Keuls test.
Eghlidi et al.
E2Effects in the Aging PrimateEndocrinology, August 2010, 151(8):3783–3794
Page 7
A subtle decrease in PDYN expression has recently
been observed in the ARC neurons of postmenopausal
women (53). Furthermore, there is evidence that PDYN
expression decreases after OVX in sheep (54) and that E2
treatment appears to enhance ARC PDYN expression in
sheep (54, 55). Although we detected a trend for PDYN
levels to decrease after OVX (experiments 2 and 3), the
results were not statistically significant. It is unclear
whether this reflects subtle differences between species or
sensitivity to effectively detect a change. It is also possible
that PDYN expression changes are more pronounced at
times of the day that were not examined in our study.
Using gene microarray analysis, we previously found that
many genes have a clear 24-h expression pattern (44, 45).
Consequently, we were concerned that the collection of
postmortem tissue in the present study may have been
performed at a suboptimal time of the day (i.e. during the
daily nadir of KNDy gene expression) and that this may
have hampered our efforts to detect changes in PDYN.
FIG. 3. Expression of KNDy in the ARC-ME of adult female rhesus
macaques across the menstrual cycle. The values represent the means
(?SEM) of KiSS-1 (A), NKB (B), and PDYN (C) during the EF, LF, and ML
phases of the cycle. No changes in the level of KNDy expression were
detected. P ? 0.05, ANOVA followed by Student-Newman-Keuls test.
FIG. 4. Expression of KNDy receptor genes in the ARC-ME of adult
female rhesus macaques during aging. The values represent the means
(?SEM) of GPR54 (A), NK3 (B), and KOR (C) from young (13.9 ? 2.1
yr), middle-aged (22.2 ? 0.4 yr), and old perimenopausal (25.8 ? 0.4
yr) animals. No changes in the level of GPR54, NK3, or KOR were
detected. P ? 0.05, ANOVA followed by Student-Newman-Keuls test.
Endocrinology, August 2010, 151(8):3783–3794 endo.endojournals.org
Page 8
Therefore, experiment 5 was designed to compare the ex-
pression of ARC genes during the middle of the day to the
expression occurring during the middle of the night. Al-
though limited in its scope, the experiment nevertheless
failed to disclose any significant diurnal differences in
been a major factor in our failure to detect changes in
expression. It should also be noted that
the present study does not rule out the
lational, rather than at the transcrip-
ative findings, we need to be cautious
before concluding that E2has no regu-
latory effect on dynorphin A in the pri-
mate ARC.
In addition to examining KNDy
genes, the present study also compre-
hensively examined the effects of age
and E2on the ARC-ME expression of
KNDy-associated receptor genes. The
expression of GPR54, NK3, and KOR
was similar in young, middle-aged, and
old perimenopausal animals and also
did not change after experimental ma-
nipulation of the sex-steroid environ-
ment or show changes across the men-
strual cycle. Hypothalamic GPR54
expression has been reported to in-
crease in postmenopausal macaques
but not in young adult animals after
OVX (48). These findings, combined
with the present results suggest that
GPR54 expression in the macaque hy-
pothalamus increases only during very
old age and not necessarily in response
ingly, GPR54 was the only gene asso-
ciated with KNDy neuropetide path-
ways that showed an obvious diurnal
expression pattern. GPR54 levels were
significantly higher in the middle of the
day (which is when tissues for experi-
ments 1–4 were collected) than in the
middle of the night. Consequently, it is
plausible that diurnal GPR54 expres-
sion plays a role in maintaining normal
circadian functions within the hypo-
thalamus and that changes in GPR54
expression after menopause (48) con-
tribute to perturbed circadian rhythms
in the elderly. Our finding that GPR54
was more highly expressed in the middle of the animals'
subjective day (i.e. at 1300 h) than in the middle of the
subjective night (i.e. at 0100 h) may have implications for
the daytime (i.e. during the animals' subjective night),
FIG. 5. Effect of short-term (A–C) or long-term (D–F) estrogen replacement on KNDy
receptor gene expression in the ARC-ME of young adult and old female rhesus macaques.
The values represent the means (?SEM) from ovary-intact (black bars), OVX (white bars), and
OVX?E2(gray bars) animals. No changes in the level of GPR54, NK3, or KOR were detected.
P ? 0.05, ANOVA followed by Student-Newman-Keuls test.
Eghlidi et al.
E2Effects in the Aging PrimateEndocrinology, August 2010, 151(8):3783–3794
Page 9
which may not be the optimal time of day for disclosing
changes in Gpr54 expression.
Although the exact role of KNDy neuropeptides in the
primate hypothalamus is unclear, there is evidence for its
rodent and sheep studies have shown coexpression of
and nonhuman primates (1), and these neurons innervate
GnRH fibers in the ME to regulate the secretion of LH (2,
24, 27, 28, 31, 56). It has also been shown that KNDy
neurons coexpress NK3 and KOR in the rodent ARC,
effects of estrogen on GnRH pulsatile secretion via auto-
crine mechanisms (15). In primates, the transition to
menopause is associated with increased pulsatile GnRH
secretion and LH release (57, 58). In addition, E2effec-
women (59), and E2and P4treatment attenuates gonad-
otropin pulse frequency (60). Because ARC-ME KNDy
neurons express estrogen and progesterone receptors
(7–15), and show ER?-mediated negative feedback of es-
trogen on GnRH (16, 17), it is likely that in primates, as
in rodents and sheep, these neurons act at the level of
GnRH neuronal terminals to mediate steroid negative
size that this negative feedback loop is likely to be func-
tional even in old perimenopausal animals and thus
responsive to hormone replacement therapy.
A small population of kisspeptin neurons resides in the
regulation of gonadotropin secretion in the rodent and
sheep (52, 61). However, it is unlikely that these neurons
in primates, which appear to rely on neuronal circuits
mediation of the positive and negative feedback effects of
ovarian hormones (38–40, 62, 63). This conclusion is
pausal rhesus macaques, which reported significant age-
related changes in KiSS-1, GPR54, and GnRH-1 expres-
sion in the MBH but not in the preoptic area (48).
Although our study focused on age-related loss of es-
rodent experiments have also examined how positive es-
trogen feedback is mediated to GnRH neurons via Kiss-1
in the AVPV (37). However, this AVPV kisspeptin neu-
there is no evidence that it also exists in sheep, nonhuman
FIG. 6. Expression of KNDy receptor genes in the ARC-ME of adult
female rhesus macaques across the menstrual cycle. The values
represent the means (?SEM) of GPR54 (A), NK3 (B), and KOR (C) during
the EF, LF, and ML phases of the cycle. No changes in the level of
GPR54, NK3, or KOR were detected. P ? 0.05, ANOVA followed by
Student-Newman-Keuls test.
TABLE 2. Diurnal differences in gene expression in the
rhesus macaque ARC-ME
0100 h 1300 h
0.35 ? 0.18
0.24 ? 0.17
0.59 ? 0.12
0.09 ? 0.01
0.05 ? 0.04
1.10 ? 0.40
1.58 ? 0.96
1.82 ? 0.75
0.61 ? 0.28
0.40 ? 0.10a
1.25 ? 0.51
0.45 ? 0.13
Values represent the mean ? SEM from three to four ovariectomized
female adults and are expressed relative to the expression of RPL13A.
aP ? 0.05.
Endocrinology, August 2010, 151(8):3783–3794 endo.endojournals.org
Page 10
of the sheep during the E2-induced LH-surge (52), but it
remains to be determined whether a similar positive feed-
back mechanism also exists in primates.
cycles and declining levels of circulating E2(46). The
present study further elucidates the role of negative feed-
to contribute to age-associated neuroendocrine alter-
ations within the hypothalamic ARC-ME. The primary
sion, which was already detectable during the perimeno-
from the ovaries starts to decline (46). No corresponding
genes were observed, suggesting that the primary modu-
lation of the hypothalamic KNDy pathways is mediated
through changes in expression of the ligands rather than
the receptors.
Support Core, Yibing Jia and Corey Singleton of the ONPRC
Molecular and Cell Biology Core, and Sharon Kryger of the
Dr. Ov Slayden for his help with the uterine histology.
185th Avenue, Beaverton, Oregon 97006. E-mail: urbanski@
This work was supported by National Institutes of Health
Grants T32 AG-023477, R01 AG-029612, U54 HD-018185,
R01 HD-029186, and P51 RR-000163.
D.H.E. is now a member of the Department of Behavioral
Neuroscience, Oregon Health & Science University, Portland,
Disclosure Summary: The authors have nothing to declare.
1. RanceNE2009Menopauseandthehumanhypothalamus:evidence
for the role of kisspeptin/neurokinin B neurons in the regulation of
estrogen negative feedback. Peptides 30:111–122
2. ChengG,CoolenLM,PadmanabhanV,GoodmanRL,LehmanMN
2010 The kisspeptin/neurokinin B/dynorphin (KNDy) cell popula-
tion of the arcuate nucleus: sex differences and effects of prenatal
testosterone in sheep. Endocrinology 151:301–311
3. HrabovszkyE,SteinhauserA,Baraba ´sK,ShughruePJ,PetersenSL,
Merchenthaler I, Liposits Z 2001 Estrogen receptor-? immunore-
brain. Endocrinology 142:3261–3264
4. Shivers BD, Harlan RE, Morrell JI, Pfaff DW 1983 Absence of oes-
tradiol concentration in cell nuclei of LHRH-immunoreactive neu-
rones. Nature 30:345–347
5. Herbison AE, Theodosis DT 1992 Localization of oestrogen recep-
tors in preoptic neurons containing neurotensin but not tyrosine
hydroxylase, cholecystokinin or luteinizing hormone-releasing hor-
mone in the male and female rat. Neuroscience 50:283–298
6. Skinner DC, Caraty A, Allingham R 2001 Unmasking the proges-
colocalization with gonadotropin-releasing neurons. Endocrinol-
ogy 142:573–579
7. Smith JT, Cunningham MJ, Rissman EF, Clifton DK, Steiner RA
2005 Regulation of Kiss1 gene expression in the brain of the female
mouse. Endocrinology 146:3686–3692
8. Smith JT, Dungan HM, Stoll EA, Gottsch ML, Braun RE, Eacker
SM, Clifton DK, Steiner RA 2005 Differential regulation of Kiss1
mRNA expression by sex steroids in the brain of the male mouse.
Endocrinology 146:2976–2984
9. Rance NE, McMullen NT, Smialek JE, Price DL, Young 3rd WS
gen receptor gene in the human hypothalamus. J Clin Endocrinol
Metab 71:79–85
10. Goubillon ML, Forsdike RA, Robinson JE, Ciofi P, Caraty A,
Herbison AE 2000 Identification of neurokinin B-expressing neu-
rons as an highly estrogen-receptive, sexually dimorphic cell group
in the ovine arcuate nucleus. Endocrinology 141:4218–4225
11. PillonD,CaratyA,Fabre-NysC,BruneauG2003Short-termeffect
nucleus of ewes. J Neuroendocrinol 15:749–753
12. Dellovade TL, Merchenthaler I 2004 Estrogen regulation of neu-
rokinin B gene expression in the mouse arcuate nucleus is mediated
by estrogen receptor ?. Endocrinology 145:736–742
13. Franceschini I, Lomet D, Cateau M, Delsol G, Tillet Y, Caraty A
arcuate nucleus co-express estrogen receptor ?. Neurosci Lett 401:
14. Gottsch ML, Navarro VM, Zhao Z, Glidewell-Kenney C, Weiss J,
Jameson JL, Clifton DK, Levine JE, Steiner RA 2009 Regulation of
and nonclassical estrogen receptor pathways. J Neurosci 29:9390–
15. Navarro VM, Gottsch ML, Chavkin C, Okamura H, Clifton DK,
Steiner RA 2009 Regulation of GnRH secretion by Kiss1/dynor-
phin/neurokinin B neurons in the arcuate nucleus of the mouse.
J Neurosci 29:11859–11866
16. Dorling AA, Todman MG, Korach KS, Herbison AE 2003 Critical
role for estrogen receptor ? in negative feedback regulation of go-
nadotropin-releasing hormone mRNA expression in the female
mouse. Neuroendocrinology 78:204–209
17. Glidewell-Kenney C, Hurley LA, Pfaff L, Weiss J, Levine JE, Jameson
JL 2007 Nonclassical estrogen receptor ? signaling mediates negative
feedback in the female mouse reproductive axis. Proc Natl Acad Sci
18. de Roux N, Genin E, Carel JC, Matsuda F, Chaussain JL, Milgrom
19. Seminara SB, Messager S, Chatzidaki EE, Thresher RR, Acierno Jr
AG, Zahn D, Dixon J, Kaiser UB, Slaugenhaupt SA, Gusella JF,
O'Rahilly S, Carlton MB, Crowley Jr WF, Aparicio SA, Colledge
WH 2003 The GPR54 gene as a regulator of puberty. N Engl J Med
20. Topaloglu AK, Reimann F, Guclu M, Yalin AS, Kotan LD, Porter
KM, Serin A, Mungan NO, Cook JR, Ozbek MN, Imamoglu S,
Akalin NS, Yuksel B, O'Rahilly S, Semple RK 2009 TAC3 and
TAC3R mutations in familial hypogonadotropic hypogonadism re-
Eghlidi et al.
E2Effects in the Aging PrimateEndocrinology, August 2010, 151(8):3783–3794
Page 11
veal a key role for neurokinin B in the central control of reproduc-
tion. Nat Genet 41:354–358
21. Han SK, Gottsch ML, Lee KJ, Popa SM, Smith JT, Jakawich SK,
Clifton DK, Steiner RA, Herbison AE 2005 Activation of gonado-
tropin-releasing hormone neurons by kisspeptin as a neuroendo-
crine switch for the onset of puberty. J Neurosci 25:11349–11356
22. Seminara SB 2005 Metastin and its G protein-coupled receptor,
GPR54: critical pathway modulating GnRH secretion. Front Neu-
roendocrinol 26:131–138
23. Shahab M, Mastronardi C, Seminara SB, Crowley WF, Ojeda SR,
Plant TM 2005 Increased hypothalamic GPR54 signaling: a poten-
Sci USA 102:2129–2134
24. ClarksonJ,HerbisonAE2006Postnataldevelopmentofkisspeptin
neurons in mouse hypothalamus; sexual dimorphism and projec-
tions to gonadotropin-releasing hormone neurons. Endocrinology
25. KeenKL,WegnerFH,BloomSR,GhateiMA,TerasawaE2008An
luteinizing hormone-releasing hormone-1 release in the stalk-me-
dian eminence of female rhesus monkeys in vivo. Endocrinology
26. Messager S, Chatzidaki EE, Ma D, Hendrick AG, Zahn D, Dixon J,
Thresher RR, Malinge I, Lomet D, Carlton MB, Colledge WH,
Caraty A, Aparicio SA 2005 Kisspeptin directly stimulates gonado-
Proc Natl Acad Sci USA 102:1761–1766
27. Krajewski SJ, Anderson MJ, Iles-Shih L, Chen KJ, Urbanski HF,
Rance NE 2005 Morphological evidence that neurokinin B modu-
lates gonadotropin-releasing hormone secretion via neurokinin 3
receptors in the rat median eminence. J Comp Neurol 489:372–386
28. RamaswamyS,GuerrieroKA,GibbsRB,PlantTM2008Structural
interactions between kisspeptin and GnRH neurons in the medio-
revealed by double immunofluorescence and confocal microscopy.
Endocrinology 149:4387–4395
29. ClarksonJ,d'AnglemontdeTassignyX,MorenoAS,ColledgeWH,
Herbison AE 2008 Kisspeptin-GPR54 signaling is essential for pre-
ovulatory gonadotropin-releasing hormone neuron activation and
the luteinizing hormone surge. J Neurosci 28:8691–8697
30. Sandoval-Guzma ´nT,RanceNE2004Centralinjectionofsenktide,
an NK3 receptor agonist, or neuropeptide Y inhibits LH secretion
amus. Brain Res 1026:307–312
31. Foradori CD, Amstalden M, Goodman RL, Lehman MN 2006 Co-
localisation of dynorphin A and neurokinin B immunoreactivity in
the arcuate nucleus and median eminence of the sheep. J Neuroen-
docrinol 18:534–541
32. Schulz R, Wilhelm A, Pirke KM, Gramsch C, Herz A 1981 ?-En-
dorphin and dynorphin control serum luteinizing hormone level in
immature female rats. Nature 294:757–759
33. KinoshitaF,NakaiY,KatakamiH,ImuraH1982Suppressiveeffect
of dynorphin-(1-13) on luteinizing hormone release in conscious
castrated rats. Life Sci 30:1915–1919
34. ChavkinC,JamesIF,GoldsteinA1982Dynorphinisaspecificendog-
enous ligand of the kappa opioid receptor. Science 215:413–415
35. Wise PM, Smith MJ, Dubal DB, Wilson ME, Rau SW, Cashion AB,
Bo ¨ttner M, Rosewell KL 2002 Neuroendocrine modulation and
repercussions of female reproductive aging. Recent Prog Horm Res
36. Downs JL, Wise PM 2009 The role of the brain in female repro-
ductive aging. Mol Cell Endocrinol 299:32–38
37. Lederman MA, Lebesgue D, Gonzalez VV, Shu J, Merhi ZO, Etgen
AM, Neal-Perry G 2010 Age-related LH surge dysfunction corre-
lates with reduced responsiveness of hypothalamic anteroventral
back in middle-aged rats. Neuropharmacology 58:314–320
38. Krey LC, Butler WR, Knobil E 1975 Surgical disconnection of the
medial basal hypothalamus and pituitary function in the rhesus
monkey. I. Gonadotropin secretion. Endocrinology 96:1073–1087
39. Knobil E 1980 The neuroendocrine control of the menstrual cycle.
Recent Prog Horm Res 36:53–88
40. Plant TM, Krey LC, Moossy J, McCormack JT, Hess DL, Knobil E
lactin secretion in the female rhesus monkey (Macaca mulatta). En-
docrinology 102:52–62
41. Brenner RM, Carlisle KS, Hess DL, Sandow BA, West NB 1983
Morphology of the oviducts and endometrium of cynomolgus ma-
caques during the menstrual cycle. Biol Reprod 29:1289–1302
42. KohamaSG,BetheaCL1995Steroidregulationoftyrosinehydrox-
ylase messenger ribonucleic acid in dopaminergic subpopulation of
monkey hypothalamus. Endocrinology 136:1790–1800
43. Noriega NC, Eghlidi DH, Garyfallou VT, Kohama SG, Kryger SG,
Urbanski HF 2010 Influence of 17?-estradiol and progesterone on
GABAergic gene expression in the arcuate nucleus, amygdala, and
hippocampus of the rhesus macaque. Brain Res 1307:28–42
44. Urbanski HF, Noriega NC, Lemos DR, Kohama SG 2009 Gene
expression profiling in the rhesus macaque: experimental design
considerations. Methods 49:26–31
45. LemosDR,DownsJL,UrbanskiHF2006Twenty-four-hourrhyth-
mic gene expression in the rhesus macaque adrenal gland. Mol En-
docrinol 20:1164–1176
46. DownsJL,UrbanskiHF2006Neuroendocrinechangesintheaging
Reprod 75:539–546
47. Rometo AM, Krajewski SJ, Voytko ML, Rance NE 2007 Hyper-
trophy and increased kisspeptin gene expression in the hypotha-
lamic infundibular nucleus of postmenopausal women and ovari-
ectomized monkeys. J Clin Endocrinol Metab 92:2744–2750
48. Kim W, Jessen HM, Auger AP, Terasawa E 2009 Postmenopausal
increase in KiSS-1, GPR54, and luteinizing hormone releasing hor-
monkeys. Peptides 30:103–110
49. Rance NE, Young 3rd WS 1991 Hypertrophy and increased gene
expression of neurons containing neurokinin-B and substance-P
messenger ribonucleic acids in the hypothalami of postmenopausal
women. Endocrinology 128:2239–2247
50. Abel TW, Voytko ML, Rance NE 1999 The effects of hormone
replacement therapy on hypothalamic neuropeptide gene expres-
sion in a primate model of menopause. J Clin Endocrinol Metab
51. Sandoval-Guzma ´nT,StalcupST,KrajewskiSJ,VoytkoML,Rance
NE 2004 Effects of ovariectomy on the neuroendocrine axes regu-
lating reproduction and energy balance in young cynomolgus ma-
caques. J Neuroendocrinol 16:146–153
52. Smith JT, Li Q, Pereira A, Clarke IJ 2009 Kisspeptin neurons in the
ovine arcuate nucleus and preoptic area are involved in the preovu-
latory luteinizing hormone surge. Endocrinology 150:5530–5538
53. Rometo AM, Rance NE 2008 Changes in prodynorphin gene ex-
pression and neuronal morphology in the hypothalamus of post-
menopausal women. J Neuroendocrinol 20:1376–1381
54. Foradori CD, Goodman RL, Adams VL, Valent M, Lehman MN
2005 Progesterone increases dynorphin a concentrations in cere-
brospinal fluid and preprodynorphin messenger ribonucleic acid
levels in a subset of dynorphin neurons in the sheep. Endocrinology
55. GoodmanRL,LehmanMN,SmithJT,CoolenLM,deOliveiraCV,
Jafarzadehshirazi MR, Pereira A, Iqbal J, Caraty A, Ciofi P, Clarke IJ
dynorphin A and neurokinin B. Endocrinology 148:5752–5760
56. Smith JT, Coolen LM, Kriegsfeld LJ, Sari IP, Jaafarzadehshirazi
MR, Maltby M, Bateman K, Goodman RL, Tilbrook AJ, Ubuka T,
Bentley GE, Clarke IJ, Lehman MN 2008 Variations in kisspeptin
and RFamide-related peptide (RFRP) expression and terminal con-
Endocrinology, August 2010, 151(8):3783–3794endo.endojournals.org
Page 12
novel medium for seasonal breeding in the sheep. Endocrinology
57. Woller MJ, Everson-Binotto G, Nichols E, Acheson A, Keen KL,
Bowers CY, Terasawa E 2002 Aging-related changes in release of
growth hormone and luteinizing hormone in female rhesus mon-
keys. J Clin Endocrinol Metab 87:5160–5167
58. Gore AC, Windsor-Engnell BM, Terasawa E 2004 Menopausal in-
creases in pulsatile gonadotropin-releasing hormone release in a non-
human primate (Macaca mulatta). Endocrinology 145:4653–4659
59. Gill S, Lavoie HB, Bo-Abbas Y, Hall JE 2002 Negative feedback
effects of gonadal steroids are preserved with aging in postmeno-
pausal women. J Clin Endocrinol Metab 87:2297–2302
60. Gill S, Sharpless JL, Rado K, Hall JE 2002 Evidence that GnRH
decreases with gonadal steroid feedback but increases with age in
postmenopausal women. J Clin Endocrinol Metab 87:2290–2296
61. Neal-Perry G, Lebesgue D, Lederman M, Shu J, Zeevalk GD, Etgen
AM, 2009 The excitatory peptide kisspeptin restores the luteinizing
hormone surge and modulates amino acid neurotransmission in the
62. Kesner JS, Wilson RC, Kaufman JM, Hotchkiss J, Chen Y,
Yamamoto H, Pardo RR, Knobil E 1987 Unexpected responses of
the hypothalamic gonadotropin-releasing hormone "pulse genera-
tor" to physiological estradiol inputs in the absence of the ovary.
Proc Natl Acad Sci USA 84:8745–8749
63. O'Byrne KT, Chen MD, Nishihara M, Williams CL, Thalabard JC,
Hotchkiss J, Knobil E 1993 Ovarian control of gonadotropin hor-
key: duration of the associated hypothalamic signal. Neuroendo-
crinology 57:588–592
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Wellcome Trust and MRC can opt for open access
Eghlidi et al.
E2Effects in the Aging PrimateEndocrinology, August 2010, 151(8):3783–3794


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We surely appreciate any Ubuntu tips that come our way. If you are an author of an app that you would like an article written about, or maybe you want to tell us about a new Gnome theme or can design some awesome wallpapers, feel free to email a tip or just to say hi! We would love to hear from you & promise to return every email.
You can contact us at:
contact-info [at] iheartubuntu [dot] com
Contact and Ubuntu Tips
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on Sunday, May 23, 2010


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The Perry and Spickett 2015 will be held on 6th and 7th June at Whitchuch Rugby Club.
Please send applications to represent the Western Area at the Perry (Open) and Spickett (Ladies) in pairs to WWBA Secretary, Jennifer Wardell firstname.lastname@example.com by 20th January 2015.
The Celtic Congress 2015 will be held at the Fishguard Bay Hotel on 29th - 31st May 2015. The Congress will be directed by the very popular Diarmuid Reddan.
Click here for brochure
Click Here for the Draft details of the new Teams League in West Wales
Click here for latest results 28/11/14
Looking for something to do on Saturday evenings?
Swansea Bridge Club now holds a 'Dragon' night on the 1st and 3rd Saturdays of the month at 7p.m. All are welcome - £4 with prizes.
From October 2014 Area Competitions and National Qualifying Events will be held at Tumble Memorial Hall at 11 a.m. The Hall is approximately one mile from the Cross Hands roundabout and will be easily accessible from both the West and East of the Area. There is a car park directly opposite the Hall.
The WWBA is still looking for a volunteer to take on the post of A.T.O. In the meantime please contact the Secretary, Jennifer Wardell, with your entries. email@example.com
If you are visiting West Wales - our beautiful land of coast and castles - then hopefully the site will help you find a bridge venue anywhere from just south of Bridgend up to Newport Pembs. You will be made very welcome at any of our Area Clubs, - especially if you are over-tired and worn out by sun, sea, cliffs and hill walking.
Congratulations to our Area winners Tony Haworth and Rob Charlesworth and we are very pleased to see that they are also the National winners in the Qualifiers.
For the National results please go to http://news.welshbridgeunion.org
Congratulations to the winners Chris Morley & Jennifer Wardell and to the runners up Bob Alderdice & Eleanor Price.
Congratulations to the winners Tony Haworth & Rob Charlesworth and to the very close runners up Wendy Thornton & Wally Heaton.
It was an enjoyable competition, and the new venue and the hot meal proved to be popular.
Congratulations to Beryl Warner and Lee Collier who were the winners of the Area Ladies' Pairs Final. Congratulations also to the runners up, Marjorie Morris and Linda Pinnock, and a big thank you to Diana Panniers for running the event for us.
Full results appear on the Swansea Bridge Club website www.swanseabridgeclub.org.uk


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I have found this wort chiller in a local shop.. I am just wondering how it work....I hook it up to the water source and put it in the wort? If yes, then one thing concerns me.....how can I use water in this chiller that is from another vessel (a vessel full of icy water)? Is there a way to make gravity push the icy water through it and thus shortening the cooling time?
You can either hook it up to your home water supply via garden hose connections or if you are using a big tub of ice water, you would need a pump to push the ice water through the chiller.
You can hook it up to a garden hose or faucet and then have a drain hose or you can buy a submersible pump to pump ice water from another vessel. I live in Minnesota and our tap water rarely gets above 60 degrees so it cools very nicely hooked up to a garden hose.
This might be the beer talking....but I really love beer!
U could also use it the opposite way if u have a pump.. Put that in a large bucket full of ice and add some water and some salt to keep it extra cold. then pump the wort from ur BK thru it and into the carboy.
With that one you hook up one end to a cold water source and the other end drains - into a sink, yard... I made mine. I have two coils from Home Depot. One is smaller than the other so I can put both in the wort. or I can put the smaller one in a bucket of cold water and ice. The big trick is to keep the wort moving. If it is still the wort immediately next to the chiller cools but it takes a long time to chill the rest. I plan someday to get an electric motor the will turn slowly and add something to stir the wort.
I use 60lb of ice to chill 10 gal of wort. I put 5 gal of water in it and use an immersion chiller. I pump the water in and put the output water into my HLT for after brew cleaning. As needed I pour return water over the ice to keep chilled water on hand. Works pretty good, 15 mins an I'm at pitching temps, plus I got plenty of water to heat and clean with. pb
I use the outlet water from my chiller to sanitize my tools and fermenting vessel. I add 1 oz of Star-san, and turn the chiller on slow. The outlet tube goes into the airlock grommet and the lid is loosely set on top of the bucket. When foam appears the chill is close to done and I have a fresh clean batch of sanitizer right where I need it for long enough to work well. I throw my spoon and anything else like the airlock, thermometer and whatever tools I need in the bucket as it is filling. You can also use an inexpensive pond or hydroponics pump to push the ice cold water through the chiller too. If I use Ice water I add a bit of salt to keep the temp below 32 and still make a batch of sanitizer with the outflow too. Wheelchair Bob
What do you mean "no Kidneys"???, WTF now I gotta drink less beer... Join the Automation sub forum in Electric brewing for a discussion of components and control systems. I did!!!!


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Angela, superb idea! My husband's record file was lost in transit after the fire. But because he had a set of promotion orders with serial numbers he was able to retrieve other documents from other files that listed his name. The Records Center is now rebuilding his file from those other documents. Thanks for getting the word out. American and personnel history is vital. Notify Administrator about this message?
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