text: tag:blogger.com,1999:blog-121780022018-10-08T04:01:34.229-05:00Penile Code AvengerPenileCodeAvengerhttp://email@example.com:blogger.com,1999:blog-12178002.post-1963356118685183952014-05-25T14:52:00.002-05:002014-05-25T14:52:41.987-05:00#YesAllWomenIn case you haven't heard of it yet, #YesAllWomen is blowing up Twitter! Details on what set off the conversation can be found here and here.Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-61632994281270832952013-08-08T09:20:00.000-05:002013-08-08T09:20:00.207-05:00Tough LoveQueendom's study of 1,234 parents who took their Parenting Style Test reveals that healthy parent-child relationships and well-adjusted children require a different view of "tough love." "Authoritative parents are able to create a balance. They don't over-compensate, like Permissive parents, and unlike Authoritarian parents, they can be strict without being harsh," explains Dr. Ilona Jerabek, Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-58788081294576361882013-04-20T02:57:00.002-05:002013-04-20T02:57:27.596-05:00April Is Sexual Assault Awareness MonthApril is Sexual Assault Awareness Month, so please take the time to educate yourself so that you can educate the children in your life. Slip Of A Girlhttps://email@example.com:blogger.com,1999:blog-12178002.post-53294432197196017302013-02-24T04:17:00.002-06:002013-02-24T04:45:59.011-06:00The More Things Change, The More They Stay The Same: Let's Talk About Domestic ViolenceNobody wants to talk about domestic violence; but somebody has to. Before we begin, here's a reminder; read it, heed it, before you comment or I'll delete it. In the news... At The Daily Beast, Barbie Latza Nadeau writes about Men Who Hate Women. While this article focuses on the increase in domestic violence against women in Southern Europe, there's, again, the reminder that the most dangerousPenileCodeAvengerhttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-23143737086459468712012-06-09T22:42:00.000-05:002012-06-09T22:42:36.632-05:00Making Noise Making Noise Sometimes reading the paper is just not good for one's blood pressure. I bought today's paper and right at the top was a headline "Googling lands in court". Ok, Google always seems to be in some kind of stir so curiosity being one of my traits I started reading. The first line stopped me cold. It read, "In a first-of-its-kind ruling, a … County Judge has ordered that a young Rickhttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-6953404027698944882012-05-16T12:42:00.000-05:002013-02-24T02:45:56.423-06:00Links & News Regarding Abuse & Domestic ViolenceMainly I provide (or curate) links to news at Snip.It. Collections pertaining to this site can be primarily found in the following collections: Feminism, Herstory, Crimes Against Humanity, and Kinsanity. Here are just a few of the recent stand outs: Science exposes the fact that violence & neglect actually changes the DNA of children. If you want to understand the differences between Deanna Dahlsadhttps://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-20000713506813970632012-05-15T13:24:00.001-05:002012-05-15T13:24:08.973-05:00Stop The Fake VAWA!The republican's H.R. 4970 is not a real Violence Against Women Act. It's not only incredibly exclusionary but puts survivors at risk! Find out how you can take action before the vote on May 16, 2012.Slip Of A Girlhttps://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-69169270510799675432012-04-20T03:43:00.001-05:002012-04-20T03:43:54.490-05:00Study Finds Reasons Why Victims & Survivors Are Stigmatized & Victimized TwiceRecent research on how families communicate when faced with serious health issues brought on by "slow moving technological disasters," like environmental disasters, may shed some insight into what happens to families and friendships after domestic violence, abuse and/or other assaults are shared. While this study specifically focused on families dealing with asbestos-related disease, I think Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-17307977171075201852012-04-18T10:02:00.002-05:002012-04-18T10:02:02.109-05:00Domestic Violence, Abuse & The EconomyAlessia's back blogging: What We Can Learn From Teen Dating Violence.Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-61624261241421089102012-03-18T13:29:00.001-05:002012-03-18T13:29:00.447-05:00Get InvolvedSee the facts and read the list here. Deanna Dahlsadhttps://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-67511683788882752722012-03-13T23:54:00.000-05:002012-03-13T23:54:47.708-05:00Dear SonBecause of the violence in our family at the hand of your father, I never really know what to do, what to believe, when you act funny and strange when I ask you about the scratches and bruises on your body... Are you just a boy uncomfortable with his mother's mothering? Or are you hiding something deeper... None of them look like hand prints or fists, but still, I worry.Slip Of A Girlhttps://email@example.com:blogger.com,1999:blog-12178002.post-8117655051100101672011-12-16T01:20:00.002-06:002011-12-16T01:27:33.504-06:00This Has Got To Stop!I opened the local paper today and the headline article read "1 in 5 women hurt by rape". Now before some scoffmiester starts whining that it's just another inflammatory bogus survey by a biased group, keep reading. The survey was run by the CDC with the help of the National Institute of Justice and the Department of Defence. The article goes on to say that 1 in 4 were the subject of domestic Rickhttp://email@example.com:blogger.com,1999:blog-12178002.post-16165488209499044962011-11-30T18:16:00.001-06:002011-11-30T18:16:56.011-06:00The Holidays For Separated Domestic Violence FamiliesThe holidays are here. For those of us who have survived domestic violence and share children from that partnership, this is an especially trying time. Forgetting, for now the increased financial difficulties of travel, holiday gift giving costs and child support, there's the very real problem of visitation times themselves. For most of us, there are no protections; courts no longer feel Slip Of A Girlhttps://email@example.com:blogger.com,1999:blog-12178002.post-65179654459377852292011-11-15T02:40:00.003-06:002011-11-15T02:48:58.494-06:00SilenceCan you hear silence?An odd question perhaps but the idea for that question came to me several days ago. I was in a restaurant that I go to often. The woman who waited on our table that day was one of the owners. Over time we, my wife and I and the woman have become casual friends. I have met her husband who we frequently see at the restaurant also. As she came to the table with our menus I Rickhttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-10723275684419563162011-11-11T00:23:00.001-06:002011-11-11T00:25:25.910-06:00A Home must respect YouOnly the second blog and I'm changing the sequence already. I just couldn't make it work, writing about being supportive without first talking about respect. So, here we go!A Home must respect youA Home is or should be an environment of mutual support and respect. The Home is where you need to feel valued most of all. When you walk in that door you need to know that you are in a place where you Rickhttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-32487989770048246242011-10-18T01:58:00.002-05:002011-10-20T16:50:58.263-05:00What A Home Should, MUST BeWhat a Home should, MUST be. I want to start this off by making a few points about what this is and is not. These are my feelings on what is necessary to have a thriving and happy Home. I had to arrange them into individual topics just to keep me from rambling too much. In an effort to not cover the same ground over and over I should let you know what the topics will be (as of now). A Home mustRickhttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-83769886622027766192011-10-17T17:30:00.001-05:002011-10-17T17:31:05.357-05:00"There Is No Honor In This Shit!"This video is of United States Marine Corps. Sgt. Shamar Thomas from Roosevelt, NY, who recently went toe-to-toe with the New York Police Department as an activist in the Occupy Wall Street movement. Thomas voiced his opinions of the NYPD police brutality that had and has been plaguing the #OWS movement. When he gets to shouting, "There is no honor in this shit!" and saying, "It takes a cowardDeanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-59746197054806337992011-10-17T16:13:00.001-05:002011-10-17T16:14:00.931-05:00Of Jail & Wails; Sorry, No Puppy Dog TailsAt Hexletive, Hex worries about "Choosing Sides" when domestic violence hits your friends -- and you are friends with both parties. There's so much to comment on in this post... Before I begin, it should be said that Hex or Hexy is a survivor herself. Not wishing to squash or second-guess a fellow survivor, I do have to take issue with a few things. And also, to help you follow along, "M" is Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-51082163266774110212011-08-09T14:10:00.001-05:002011-08-09T14:10:11.827-05:00"Certain Things Hang On Forever"In Framing Recovery from Child Sexual Abuse, Lisa Wade discusses anti-child abuse ads which might go too far... The exhibit for discussion (above) is an Italian ad designed to draw attention to the seriousness of child sexual abuse. Does the ad send a "If you have been sexually abused, you will be broken for the rest of your life" message? I'm not a survivor or child abuse, so maybe I shouldPenileCodeAvengerhttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-18562746800522980162011-06-04T06:01:00.001-05:002011-10-17T17:33:29.945-05:00I've Been AngryI've been angry. At certain moments in your life you seem either to be faced with what's probably best described as the fight or flight response: you can give in (to others, depression, etc.) or get good and angry and fight back. I don't mean with your fists -- at least not at another person; I mean you can fight for yourself, your children, your family, your rights. As a recovering victim of Slip Of A Girlhttps://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-86833485520841023132011-05-20T07:36:00.000-05:002011-05-20T07:36:10.026-05:00Victim BashingThese are recent cases of victim bashing in cases of rape, but as domestic violence is less significant in terms of media coverage... The Latest Victim-Blamers: The New York Times? What's HIV Got to Do With It? New York Post Violates Alleged Victim's Rights in Strauss-Kahn Case Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-70934600172579457182011-04-28T06:15:00.001-05:002011-04-28T06:15:34.486-05:00Skating On Thin IceAfter a traumatic event, how you see things changes. Quite literally. When you survive domestic violence, you often aren't allowed this fact of life. Recognizing or acknowledging to others how you've been affected by the trauma(s) is a "luxury" you can't afford because the perceptions & projections -- along with observable deeds and actual words of other people already accuse you of not getting Deanna Dahlsadhttps://email@example.com:blogger.com,1999:blog-12178002.post-52971967577358732912011-03-13T14:18:00.001-05:002011-10-17T17:34:52.837-05:00A Note To My SonYou're too young to understand all this, and despite what your father does -- including you in his rants about subjects which are not for children -- I'm not going to tell you this... When you are older and can understand it all, there will be time for the realities. For now, I simply must say it. Baby, I know it's been months since I've seen you -- and it probably will be a few more. I Slip Of A Girlhttps://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-81906336643083411182011-02-10T23:02:00.003-06:002012-04-20T02:01:43.384-05:00Breaking The Terrible Painful SilenceWhen I was asked to join Penile Code Avenger, and its sister-site, More Than Surviving, Thriving, I was excited to begin. I even wrote about it at my personal blog. But when it came time to actually participate... I felt like I was on stage, on the exam table, feet in the stirrups, speculum in place, under a spotlight -- with the whole show up on the JumboTron for those folks in the cheap seats.Miss FussyPantshttp://firstname.lastname@example.com:blogger.com,1999:blog-12178002.post-16350206723914068142010-05-17T00:38:00.002-05:002011-02-10T23:48:40.337-06:00A Quick Note (UPDATED: Now It's Not So Short)I'm watching the "very special domestic violence" episode of Army Wives right now. It's 37 minutes through and I'm pretty sure that this is going to focus and end on the usual cliches. UPDATE: OK, like I thought, the whole focus of the show was on how a woman "should be big and brave enough to leave." Yeah, we've heard that message for decades; that's not the problem. The problem is what PenileCodeAvengerhttp://email@example.com


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Mumbai Crime: Duo nabbed at T2 trying to smuggle 183 live turtles
Customs officials nabbed Sayyad Ibrahim Sikandar and Shahjahan Naina Mohammed before they boarded a flight to Kuala Lumpur on Saturday night; said the turtles are worth Rs 1.83 lakh
Officials of the Air Intelligence Unit (AIU) nabbed two passengers, who were supposed to fly to Kuala Lumpur on Saturday night, and rescued 183 spotted pond turtles from their possession. The arrested smugglers were identified as Sayyad Ibrahim Sikandar and Shahjahan Naina Mohammed. Both hail from Chennai.
The Customs officials said the rescued turtles, valued at Rs 1.83 lakh, are listed under Schedule-I of the Wildlife Protection Act, 1972. The smugglers were supposed to board Malindo Air's OD 216 flight at 11:15 pm from T2. After the primary investigations, the Customs officials handed over the turtles to the Thane Forest Department officials.
The accused, both from Chennai, were nabbed at T2 before they boarded the flight to Kuala Lumpur with the turtles
"This is not the first time that a protected species has been rescued, and neither will it be the last. But what remains a matter of concern is lack of rehabilitation centres for the rescued animals. The Forest Department needs to take up this issue on a war footing. We had informed the Principal Secretary of Forests Vikas Kharge about this particular issue and he had promised to deal with it immediately.
However, it's been a month since we met him, and there seems to be no progress in this matter," said Shakuntala Mazumdar, president, Thane Society for the Prevention of Cruelty to Animals (TSPCA).
Expert speak
Pawan Sharma, founder, Resqink Association for Wildlife Welfare (RAWW) said, "The spotted pond turtle is a protected species. They deserve the same level of protection given to leopards, lions or tigers. Confiscation of such a large number is shocking. Wildlife crimes are rising with each passing day, especially against lesser-known species.
This is a matter of great concern and needs to be countered immediately." Veterinary doctor Deepa Katiyal, to whom these turtles were handed over, said, "While most of the turtles are healthy, a few need treatment because they've suffered eye injuries.
The smallest turtle weighs 100 grams, while the largest ones are about 600 grams. There is a possibility that these turtles were to be slaughtered and sold as exotic meat in Kuala Lumpur."
DISCLAIMER: mid-day and its affiliates shall have no liability for any views, thoughts and comments expressed on this article.
Martyn Ford fitness secrets

subset: commoncrawl


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The new iPad newspaper The Daily will be featured in a Super Bowl commercial today. The Daily has tweeted that The Daily will be the first app to be featured in a Super Bowl ad. AllThingsD has reported about The Daily Super Bowl commercial first. Now with the tweet it is not a rumor anymore, but fact.
Indirectly the Apple iPad will now be featured in a Super Bowl commercial today and can counter a bit the Motorola Xoom Super Bowl Commercial. The Motorola Xoom Android Tablet Super Bowl ad directly attacks the iPad.
There are no details yet about the The Daily Super Bowl commercial. Maybe it will be in tune with Apple ads, but it could also be totally different. The Daily needs some advertising as the first iPad newspaper has some usability issues according to Daring Fireball.
Just like last year, I4U News will be featuring again the Super Bowl for Geeks Guide. Of course the Super Bowl 2011 for Geeks Guide will be all about the Super Bowl Commercials and not the big game.
Watch the first 2011 Super Bowl Ads now.
The Super Bowl XLV will take place in the Dallas Cowboys stadium on Sunday, Feb. 6th 2011.
For a list of the current official and rumored Super Bowl ads for 2011 check out our Super Bowl Ads 2011 Guide.
See also our Super Bowl XLV Facts and Times Guide.


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Welcome to West Palm Beach located in Palm Beach county Florida, home of the sandy white beaches, the Professional Golfers Association and one of the world's largest wetlands, the Florida Everglades. Most travelers think you have to go all the way down to Miami to see the Everglades and thats just not the case, just 45 minutes from down town West Palm Beach lays the north eastern boundaries of the Everglades known as the Loxahatchee Preserve in Parkland.
The Florida Everglades are home to 67 different threatened or endangered species.
Our airboat rides are private tours of the everglades that run year round 7 days a week with the exception of Thanksgiving, Easter and Christmas day, our airboat tours are by appointment only.
Treat yourself or your family to an amazing airboat ride thru the River of Grass on one of our custom airboats. We offer different packages for your convenience ranging from one hour to three hour charters tailored to your needs. All of our Everglades tours are private, so who ever you bring is the only persons on the trip . We have fan boat rides as visitors refer to for everyone, couples, families and corporate groups.
Our airboat rides give you an in depth look at the vast ecosystem of the Florida Everglades as well as the aquatic plants and bird life. We cant forget the famous American alligators that we will encounter throughout the trip. We incorporate a little bit of everything into our airboat rides, you will not only have a great ride but our Captains will teach you about the echo system, history of the everglades as well as wildlife viewing.
Our airboats are safe, fun, and very comfortable with padded seats for your comfort, we spare no expense in providing some of the finest airboats in the industry with top of the line equipment and high performance motors for those of you that like to go a little faster. We cater to the customers needs if you want more echo at a slow pace or your a thrill seeker we have the boat and package to fit your needs. All of our Captains are U.S.C.G certified Captains who have grown up in the everglades and provide years of experience in the everglades. We are proud to be a family owned business that can provide one on one airboat tours with our customers.
One out of every three Floridians relies on the water supply from the Everglades.
Safety is a number one priority for our customers, all of our boats are to U.S.C.G. standards and meet all of the regulations to operate on a commercial basis, the boats are equipped with all approved safety equipment, life vests, hearing protection, and marine radios.
Airboat Rides west palm beach also can be hired for sight work, photo shoots of the everglades, biology studies and film or print work. We have worked with many photographers and producers on major film shoots in south Florida as well as government agencies for sight work or university studies. please call us for quotes for this type of work.
One of the most exciting family attractions in west palm beach is an airboat ride through loxahatchee preserve in parkland. Please read through are web-site for airboat ride packages as well as location and directions to our boat ramp. Please remember we are by appointment only and it is always best to book your everglades tour in advance by emailing us or calling us at 1-561-252-4030.


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Recapping Kentucky's 59-14 win over Tennessee-Martin at Commonwealth Stadium on Saturday.
Patrick Towles completed his four passes but the game-opening drive by UK stalled at the UT Martin 35-yard line. On UK's second drive, a 44-yard strike from Towles to Javess Blue set up a 2-yard TD run by Jojo Kemp. The UK offense looked sharp. It was 14-0 when Nebraska transfer Braylon Heard popped his first UK carry for 73 yards. The next time Heard touched the ball, he went 43 yards for a score. A 90-yard TD drive ended with 29-yard pass to freshman Blake Bone. A 91-yard drive ended with an 18-yard TD run by freshman Mikel Horton.
Halftime: Kentucky 35, Tennessee-Martin 0.
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Thumbs up: The UK offense was fast, effective and used a lot of weapons.
Thumbs down: It is being picky, but at times UK looked a little soft up the middle on defense.
UK's J.D. Harmon forced a fumble on the second-half kickoff but the Cats failed to take advantage. A Fred Tiller interception got the ball back for the Cats. This time, Towles got loose and ran 23 yards for a score, marking the first time since 2008 UK rushed for five scores. An Austin MacGinnis field goal made it 45-0. A 79-yard completion set that up, most of it coming from DeMarco Robinson after the catch. The next UK score came when freshman Kendall Randolph forced a fumble and Khalid Henderson romped 89 yards to score. It was 52-0 after three quarters.
Final: Kentucky 59, Tennessee-Martin 14.
Thumbs up: Patrick Towles: It was a nice debut for the sophomore quarterback who won the job in a close battle. He was 20-for-29 passing for 377 yards, no interceptions, 30 yards rushing and no sacks.
Thumbs down: Kentucky did not get its first shutout since 2009 (but the defense did make big plays).


text: I have done a ton of research and spent time on tuturials, so i decided that I should give it a shot and try to develop my first application. I need the user to choose whether he/she has a new or existing project, and if it is a new project, then fill out all the information. But if they have an existing project created, then they can use a drop down button in order to choose which project they wish to open, and it opens it just as they left it when they were done editing it before. I think i need to create a database but the more i try the more lost i get. If someone can steer me in the right direction it would be great. I've tried googling my problem, but i really dont know how to ask it. I hope I have given you enough information to help me. p.s. I have already created the home screen and the new project and existing project activities, but they contain no content. Im also using eclipse Thanks again Laffy


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Call me old-fashioned, but when an industry group throws a spread to promote its latest research findings, shouldn't one expect those findings to be insightful and revealing?
The Online Publisher's Association (OPA) recently unveiled its "A Sense of Place: Why Environments Matter" study. Some of the findings:
- "A site's content is strongly correlated with how its advertisers are perceived."
- "The more trusted, relevant, and timely the content, the greater the willingness to recommend the site to others."
- "The more reputable, relevant and respected are the advertised brands." (Those brands that advertise on the site).
Not particularly surprising information, if I might opine.
Don't environments matter in most everything we do? Doesn't a restaurant with a pleasant ambience attract patrons to it? Don't we want to live in a house that's aesthetically pleasing? Or drive a car that looks nice?
Environments do matter…but, as trite as it may sound, it's what's on the inside that counts. If the food at the restaurant isn't good, the patrons won't return. If the house is a shambles inside, people won't want to visit. And, although that car might be beautiful to look at, it may be uncomfortable to drive.
What I think the OPA was getting at was that online sites that offer both environment and content are the ones where its readers are more likely to have a higher regard for the site and for the advertisers who appear there. (Also inherent in all this is site navigability. A site with good content and an environment that a user finds difficult to navigate will affect the user's perception of that site.)
But, isn't that the way magazines have been positioning themselves for decades? Don't advertisers put ads for certain products in editorial environments that will favorably reflect on them? Aren't you known by the company you keep? In an attempt to spur online ad sales, it seems like the OPA was stating the obvious. But, environments and content have always mattered.
— Dave Capano, Director of Media Services


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Although microbial infections can alter steady-state hematopoiesis, the mechanisms that drive such changes are not well understood. We addressed a role for IFN-γ signaling in infection-induced bone marrow suppression and anemia in a murine model of human monocytic ehrlichiosis, an emerging tick-borne disease. Within the bone marrow of Ehrlichia muris-infected C57BL/6 mice, we observed a reduction in myeloid progenitor cells, as defined both phenotypically and functionally. Infected mice exhibited a concomitant increase in developing myeloid cells within the bone marrow, an increase in the frequency of circulating monocytes, and an increase in splenic myeloid cells. The infection-induced changes in progenitor cell phenotype were critically dependent on IFN-γ, but not IFN-α, signaling. In mice deficient in the IFN-γ signaling pathway, we observed an increase in myeloid progenitor cells and CDllbloGr1lo promyelocytic cells within the bone marrow, as well as reduced frequencies of mature granulocytes and monocytes. Furthermore, E. muris-infected IFN-γR–deficient mice did not exhibit anemia or an increase in circulating monocytes, and they succumbed to infection. Gene transcription studies revealed that IFN-γR–deficient CDllbloGr1lo promyelocytes from E. muris-infected mice exhibited significantly reduced expression of irf-1 and irf-8, both key transcription factors that regulate the differentiation of granulocytes and monocytes. Finally, using mixed bone marrow chimeric mice, we show that IFN-γ–dependent infection-induced myelopoiesis occurs via the direct effect of the cytokine on developing myeloid cells. We propose that, in addition to its many other known roles, IFN-γ acts to control infection by directly promoting the differentiation of myeloid cells that contribute to host defense.
As most mature blood cells have only a limited lifespan, hematopoiesis, under homeostatic conditions, acts to maintain the continual production of new cells. However, during immunological stress, caused by factors such as chemotherapy or microbial infection, the demand for immune cells is met through enhanced production and mobilization of blood cells. This requirement for new blood cells can, in turn, result in the proliferation and differentiation of hematopoietic progenitor cells. Although much of our basic knowledge of hematopoiesis has been obtained from studies of steady-state hematopoiesis, a variety of viral, bacterial, and fungal infections can alter the cellular composition and/or phenotype of bone marrow progenitor cells (1–3). The mechanisms underlying such changes are likely multifactorial and infection-dependent; however, the factors that regulate hematopoiesis during microbial infections have not been characterized in depth.
Studies that have addressed the effects of infections and inflammation on hematopoiesis have revealed that these processes can result in the enhanced production of granulocytes (4–8). This response, termed "emergency granulopoiesis" (4), functions to increase neutrophil output in response to infection, thereby facilitating innate host defense. Factors that promote emergency granulopoiesis include the CSFs, IL-3, and IL-6 (9–11), and the transcription factor CCAAT/enhancer-binding protein β (12). Thus, inflammation, mediated by soluble factors, promotes the production of granulocytes and augments the innate immune response. Despite knowledge on how these mature bone marrow cells can respond to infection-induced hematopoietic stress, less is known regarding how myeloid progenitor cells respond to inflammatory conditions.
IFN-γ plays a critical role in innate immunity to intracellular pathogens, including the ehrlichiae, and is essential for host defense via its ability to activate macrophages and to promote Th1 responses. A lesser known function of IFN-γ is its ability to act as a moderator of hematopoiesis. A historical paradigm has been that IFN-γ suppresses hematopoiesis, as was shown in several in vitro culture models (13–15). A role for IFN-γ in governing infection-induced myelopoiesis was first demonstrated in mycobacterial infection (16), where it was proposed that IFN-γ acts to suppress myelopoiesis. Other studies have shown that IFN-γ–induced genes are critical for hematopoietic stem cell (HSC) survival/maintenance (17, 18), also suggesting that IFN-γ plays a direct role in hematopoiesis. IFN-α, a related and important antiviral cytokine, was shown to activate dormant HSCs, demonstrating that IFNs can act directly on HSCs (19). In a model of malaria infection, it has been shown that IFN-γ is essential for the generation of a unique progenitor population that gives rise to mostly myeloid cells (20). Thus, IFN-γ is emerging as a factor in the regulation of infection- or stress-induced hematopoiesis.
To address a role for IFN-γ in infection-induced hematopoiesis, we have used a murine model of human monocytic ehrlichiosis (HME), an emerging infectious disease caused by the tick-borne pathogen Ehrlichia chaffeensis. The ehrlichiae are obligate intracellular bacteria that infect mononuclear cells (21). HME is a febrile illness and patients exhibit a severe loss in hemoglobin and platelets (21, 22). Infection of C57BL/6 mice with a closely related pathogen, Ehrlichia muris, causes hematological abnormalities, including anemia and thrombocytopenia (23), similar to what is observed in HME patients. We have proposed that the hematological changes that occur during ehrlichiosis are a consequence of alterations in hematopoiesis (23). In this study, we extend our previous studies by demonstrating that ehrlichial infection induces changes in the phenotype and function of myeloid progenitor cells and augments the production of granulocytes and monocytes, and that these changes require direct IFN-γ signaling. These studies highlight a novel and important role for this factor in host defense during bacterial infection.
Materials and Methods
The mice used in these studies were obtained from The Jackson Laboratory (Bar Harbor, ME) or were bred in the Animal Care Facility at the Wadsworth Center under microisolator conditions. C57BL/6 mice were used as a congenic control strain. The following transgenic and gene-targeted strains were used: IFN-αR–deficient mice (a gift of Dr. Susan Swain, Trudeau Institute, Saranac Lake, NY), IFN-γR1–deficient mice (B6.129S7-Ifngr1tm1Agt/J, referred to as IFN-γR–deficient mice), enhanced GFP (EGFP)-transgenic mice (C57BL/6-Tg(ACTB-EGFP)1Osb/J), and a C57BL/6 congenic strain of mice (B6.SJL-Ptprca Pepcb/BoyJ). All animal studies were performed in accordance with Wadsworth Center Animal Care and Use Committee guidelines.
Bacteria and infections
Mice were infected, via i.p. injection, between 6 and 12 wk of age, with 50,000 copies of the E. muris bacterium. The inoculum was generated from spleen mononuclear cells that were harvested from infected mice and stored at −80°C, as previously described (24). Bacterial copy number was determined by real-time quantitative probe-based PCR, as previously described (23, 25).
Flow cytometry and Abs
In vitro hematopoietic progenitor cell assays
Bone marrow cells were plated at 2.5 × 104 cells per 35-mm tissue culture dish, in duplicate, and cultured in methylcellulose media (MethoCult GF M3434; StemCell Technologies, Vancouver, BC, Canada) along with recombinant cytokines for colony assays of murine cells, following the manufacturer's instructions. Total colonies derived from granulocyte-macrophage (CFU granulocyte, macrophage) and multipotential (CFU granulocyte, erythrocyte, macrophage, megakaryocyte) progenitor cells were scored after 7 d incubation at 37°C in 5% CO2.
Generation of bone marrow chimeric mice
C57BL/6 and IFN-γR–deficient (CD45.2+) mice were lethally irradiated (950 rad administered in two doses, 3 h apart). After irradiation, the hematopoietic compartment of the irradiated mice was reconstituted with 2 × 106 bone marrow cells obtained from CD45.1 congenic mice. To generate the mixed chimeras, CD45.1+ mice were irradiated as above and reconstituted with a 1:1 ratio of bone marrow (2 × 106 cells) harvested from EGFP-expressing and IFN-γR–deficient mice (both CD45.2). After irradiation and reconstitution, all mice were administered antibiotics in their drinking water for 1 wk, and 6 wk postreconstitution, the mice were bled and screened to determine the degree of hematopoietic chimerism. The chimeric mice were infected with E. muris ∼8 wk postreconstitution.
Flow cytometric cell sorting
Promyelocytes were identified in bone marrow by low surface expression of CDllb and Gr-1, using CDllb-FITC and Gr-1-PE. Bone marrow cells were purified by flow cytometry from day 15 E. muris-infected C57BL/6 mice and IFN-γR–deficient mice, using an Aria II equipped with FACSDiva software (BD Biosciences).
Gene expression analyses
RNA was extracted from flow cytometrically sorted CDllbloGr-1lo cells by the RNeasy method (Qiagen, Valencia, CA). RNA integrity was analyzed with an Agilent bioanalyzer (Agilent Technologies, Foster City, CA), and only RNA preparations with an RNA integrity number of 8.0 or higher were used for gene expression studies. Genomic DNA was removed from RNA samples with TurboDNA-free DNAse (Ambion, Austin, TX), and cDNAs were generated with a first-strand synthesis kit (SABiosciences, Frederick, MD). Gene expression analyses were performed using an IFN-γ gene array assay (SABiosciences; APMM-064).
Statistical analyses
Statistical analyses were performed with a Student t test, using GraphPad Prism software (GraphPad Software, La Jolla, CA); a p value <0.05 was considered significant.
Type II IFN-dependent alterations in bone marrow progenitor cells during infection
E. muris infection of C57BL/6 mice provides a model in which to investigate the pathogenesis of HME (21). In C57BL/6 mice, bacterial burden is maximal between days 8 and 9 postinfection in the spleen, and it declines to very low numbers by day 30 postinfection (21, 26). E. muris causes marked anemia and thrombocytopenia, two hallmarks of HME (21, 22). We previously demonstrated that these blood abnormalities, which are maximal during the second week of infection, occur concomitantly with a decrease in bone marrow colony-forming cells (23). Additional studies revealed that infection induced the mobilization and/or depletion of B220+ lymphocytes from the bone marrow, which was coincident with an increase in the frequency of immature and mature granulocytes at this site (23). Significant changes in the cellular composition of the bone marrow begins by day 8 postinfection and persists through day 15 postinfection, which prompted us to use these time points for our analyses. In this study, we have extended our previous observations by examining hematopoietic progenitor cell populations in the bone marrow.
In uninfected mice, expression of IL-7Rα (CD127), Sca-1, and c-Kit on lineage-negative (Lin−) bone marrow cells can distinguish common lymphoid progenitors (CLPs; IL-7Rα+c-KitloSca-1+) and myeloid progenitors (IL-7Rα−c-KithiSca-1−) (27, 28). Using this criteria, we determined that E. muris infection caused a modest reduction in phenotypically defined CLPs in C57BL/6 mice: a small decrease in frequency was observed on day 8 postinfection (Fig. 1A), although the number of CLPs was unaffected (data not shown). In contrast, there was almost a complete loss of the IL-7Rα−c-KithiSca-1− myeloid progenitor cells by day 8 postinfection, and a population of Sca-1+ cells emerged.
Sca-1 expression is known to be regulated by IFN-γ signaling (29), and recent studies have implicated a role for IFNs in driving the differentiation of hematopoietic progenitor and stem cells (19, 30, 31). To determine whether IFN signaling was responsible for the loss of IL-7Rα−c-KithiSca-1− cells and the emergence of the Sca-1+ population, we examined the bone marrow of infected mice deficient for either the IFN-α or IFN-γR. In the absence of IFN-α signaling, little effect was noted on the phenotype of Lin−IL-7Rα− progenitor cells during E. muris infection, as compared with infected C57BL/6 mice (Fig. 1B). However, in the absence of IFN-γ signaling, reduced frequencies and numbers of IL-7Rα−c-KithiSca-1+ cells were observed postinfection, relative to infected C57BL/6 mice (Fig. 1C, 1D). Moreover, we observed an ∼8-fold decrease in the frequency and numbers of IL-7Rα−c-KithiSca-1− cells in both C57BL/6 and IFN-αR–deficient mice postinfection, but no significant decreases in this population in IFN-γR–deficient mice. Thus, the alterations in bone marrow progenitor cells observed during infection were dependent on IFN-γ, but not IFN-α, signaling.
Infection-induced alterations in bone marrow progenitor cells are correlated with anemia and changes in leukocyte frequencies in the peripheral blood
To further investigate the role of IFN-γ signaling on infection-induced hematopoiesis, we next examined the bone marrow progenitor cell populations in greater detail. During infection there was a paucity of Sca-1− cells in C57BL/6 mice. As Sca-1 is known to be induced by IFN signaling, we used two additional markers, CD34 and FcγRII/III (CD16/32), to characterize myeloid progenitor subsets within the Lin−IL-7Rα−c-Kithi Sca-1− and Sca-1+ fractions of cells (as shown in Fig. 1B). In uninfected mice, cells that are Lin−IL-7Rα−c-KithiSca-1− and CD34−FcγRII/III− are megakaryocyte-erythrocyte progenitors (MEPs) and the CD34loFcγRII/III− fraction are common myeloid progenitors (CMPs); the CD34+FcγRII/III+ population has granulocyte and monocyte potential and are thus referred to as granulocyte monocyte progenitors (GMPs) (28). We observed that, in mock-infected mice, expression of FcγRII/III is very low on the Lin−IL-7Rα−c-KithiSca-1+ cells but is increased during infection in both C57BL/6 and IFN-γR–deficient mice. We gated on total Lin−IL-7Rα−c-KithiSca-1− cells and quantified the numbers of classically defined MEPs, CMPs, and GMPs (Fig. 2A, 2B). We observed that the frequency and number of each of these myeloid populations was reduced during E. muris infection, with significant reductions in the frequency of CMPs and GMPs in C57BL/6 mice. A significant reduction in the number of CMPs was also noted in IFN-γR–deficient mice. We next examined the frequency and numbers of infection-induced Sca-1+ MEPs, CMPs, and GMPs (Fig. 2C, 2D). As Sca-1 is not normally expressed on MEPs, CMPs, and GMPs, each population in this study is referred to as "infection-induced." The numbers of infection-induced MEPs, CMPs, and GMPs were significantly increased in C57BL/6 mice during infection, whereas IFN-γR–deficient mice exhibited an increase in the CMP population only. The increase in Sca-1+ myeloid progenitor cells mirrored the loss of Sca-1− cells, which suggested that the surface expression of Sca-1 was induced on the classically defined myeloid progenitors, as was previously documented during bacterial infection (2). Thus, IFN-γ signaling contributed to the infection-induced loss of classically defined myeloid progenitors and the emergence of a Sca-1+ population of progenitor cells.
Because the identification of myeloid progenitor cell subsets relies on Sca-1 expression, which can be altered during inflammation, we next evaluated bone marrow colony formation to determine whether myeloid progenitor cells were altered in the absence of IFN-γ–dependent signaling. E. muris infection suppresses bone marrow colony formation (23), which is consistent with the reduced numbers of classically defined myeloid progenitors that we have observed. In contrast to C57BL/6 mice, E. muris infection did not inhibit colony forming activity in IFN-γR–deficient mice; in fact, bone marrow harvested from E. muris infected IFN-γR–deficient mice exhibited increased colony formation in vitro (Fig. 3A). These data suggest that the IFN-γ–dependent changes in the phenotype of myeloid progenitor cells observed during infection are associated with functional differences in these bone marrow cells.
We next addressed whether the alterations in progenitor populations within the bone marrow correlated with changes in the peripheral blood during infection. Hemoglobin levels began to diminish on day 8 postinfection, and they were significantly reduced by day 15 postinfection in C57BL/6 mice but not in IFN-γR–deficient mice (Fig. 3B). Platelet numbers were also reduced on days 8 and 15 postinfection in C57BL/6 mice, but platelets did not decrease substantially in the IFN-γR–deficient mice until day 15 postinfection (Fig. 3C). In C57BL/6 mice, hemoglobin concentration and platelet numbers were restored to nearly normal levels by day 30 postinfection (23). Thus, in IFN-γR–deficient mice the onset of anemia and thrombocytopenia occurred around the time of death in infected mice. These data reveal that thrombocytopenia was associated with reduction or loss of classically defined MEPs in C57BL/6 mice but not in IFN-γR–deficient mice, and they suggest that the loss of the progenitor cell population is at least in part responsible for the decline in platelet number in the periphery.
We also observed changes in the frequency of WBC populations in the peripheral blood of infected mice. Most striking was the observation that infection induced an apparent expansion of blood monocytes and neutrophils, as well as a contraction of lymphocytes, in the C57BL/6 mice (Fig. 3D). In contrast, IFN-γR–deficient mice exhibited a much reduced expansion of the blood monocytes, but increased neutrophils. These data are consistent with our hypothesis that the observed change in progenitor cell phenotype during infection is a consequence of IFN-γ–dependent changes in hematopoietic progenitor cell function and altered myeloid differentiation.
Altered infection-induced myelopoiesis in mice lacking IFN-γ signaling
During E. muris infection, the bone marrow becomes dedicated to the production of granulocytes (23). To determine whether the IFN-γ–dependent change in the number and phenotype of myeloid progenitors had a direct impact on the differentiation of myeloid cells, we examined the bone marrow of C57BL/6 and IFN-γR–deficient mice for its ability to support granulocyte production and differentiation. To determine the maturation status of the bone marrow granulocytes, we monitored cell-surface expression of CD11b and Gr-1 (Fig. 4A). Promyelocytes express intermediate levels of both cell surface molecules (i.e., are CDllbloGr-1lo), whereas immature neutrophils are CDllbloGr-1hi and mature neutrophils are CDllbhiGr-1hi (7). The maturation status of these populations from mock and infected mice was confirmed cytologically (data not shown).
During the first 2 wk of infection, the frequency of granulocytes in the bone marrow increased from ∼40% to nearly 50% in C57BL/6 mice (Table I). However, due to the decrease in bone marrow cellularity during infection (Ref. 23 and data not shown), the change in number of total granulocytes was not significantly different between uninfected and infected mice. We observed a significant decrease in the number of CDllbloGr-1lo cells in C57BL/6 mice on day 8 postinfection, although the numbers returned to normal by day 15 postinfection (Fig. 4B). In contrast, the frequency of granulocytes increased from 37% to nearly 80% in the bone marrow of IFN-γR–deficient mice (Table I). In infected IFN-γR–deficient mice, we observed a significant increase in the numbers of CDllbloGr-1lo promyelocytes in the bone marrow by day 15 postinfection (Fig. 4B). In the infected IFN-γR–deficient mice, the CDllbloGr-1lo promyelocytic cell population constituted more than half of all cells within the bone marrow. The apparent expansion of the CDllbloGr-1lo cells was only noted during infection, as uninfected C57BL/6 and IFN-γR–deficient mice exhibited very similar frequencies of granulocytes at each developmental stage. It is possible that the infection-induced expansion of the CDllbloGr-1lo cells could have resulted from increased proliferation, decreased cell death, and/or a block in differentiation. Infected IFN-γR–deficient mice had fewer B220+ B cells in the bone marrow, as was observed in C57BL/6 mice (Ref. 23 and data not shown), indicating that the infection-induced alterations in bone marrow B lymphocytes were unaffected by the loss of IFN-γ signaling. Thus, our data indicate that IFN-γ signaling is required for myeloid, but not lymphoid, differentiation during infection.
The infection-induced alterations in the bone marrow were reflected in the periphery, as infection increased the frequency and number of granulocytes in the spleens of both C57BL/6 and IFN-γR–deficient mice (Fig. 4C, 4D, Table I). Neutrophils normally represent a small fraction (<2%) of the cells in the spleens of both C57BL/6 and IFN-γR–deficient mice. However, E. muris infection induced a 1.5- to 2-fold increase in the frequency of spleen granulocytes in C57BL/6 mice, but this corresponded to a 10- to 12-fold increase in the numbers of CDllbloGr-1lo and CDllbloGr-1hi cells due to splenomegaly (Fig. 4D). This difference was more pronounced in infected IFN-γR–deficient mice, which exhibited an ∼40-fold increase in the number of CDllbloGr-1lo promyelocytic cells, relative to uninfected mice; ∼15% of the cells in the spleens of IFN-γR–deficient mice were CDllbloGr-1lo cells. Thus, ehrlichial infection causes an expansion of immature and mature granulocyte populations, and these changes are greatly exacerbated in the absence of IFN-γ signaling. These findings are consistent with related studies of Mycobacterium-infected mice (16).
Cytological examination of neutrophils in spleen samples from the infected mice revealed that the splenic neutrophils in the IFN-γR–deficient mice, but not C57BL/6 mice, harbored inclusions containing intracellular ehrlichiae (Fig. 4E). This observation was unexpected, as the ehrlichiae are largely monocytotropic and have never been reported to infect neutrophils (22). Given that many more phenotypically immature neutrophils were detected in the spleens of IFN-γR–deficient mice, the altered ehrlichial cell tropism in the IFN-γR–deficient mice may also explain the higher degree of bacterial infection. We interpret the altered bacterial tropism in the IFN-γR–deficient mice to suggest that neutrophils that develop during infection, in the absence of IFN-γ signaling, can phagocytose but not kill the intracellular bacteria. An alternative explanation is that the cells identified as neutrophils in the spleens of infected IFN-γR–deficient mice have some characteristics of monocytes, making them permissible to infection with E. muris; however, these possibilities remain to be addressed.
A direct role for IFN-γ signaling in infection-induced hematopoiesis
It was possible that the persistence of myeloid progenitors, as well as the enhanced granulopoietic response observed in the absence of IFN-γ signaling, was mediated indirectly, perhaps by signaling on nonhematopoietic cells within the bone marrow. To address whether our observations were due to direct IFN-γ signaling on hematopoietic cells, we first generated bone marrow chimeric mice by irradiating IFN-γR–deficient and C57BL/6 mice and reconstituting them with congenic wild-type bone marrow. Eight weeks post-reconstitution, the chimeric mice were infected with E. muris and bone marrow cells were analyzed on day 8 postinfection for progenitor cell phenotype. We found that the increased frequency of Lin−c-Kit+Sca-1+ cells that emerged in infected wild-type mice was observed in both C57BL/6 and IFN-γR–deficient recipients (Fig. 5), supporting the idea that IFN-γ acts directly on hematopoietic cells, and not indirectly, via interaction with nonhematopoietic stromal bone marrow cells.
It was possible that the enhanced granulopoietic response observed in the absence of IFN-γ signaling could also be a consequence of higher bacterial infection and inflammation in the IFN-γR–deficient hosts. To address this concern we next generated radiation-induced mixed bone marrow chimeric mice. Irradiated congenic mice were reconstituted with a 1:1 ratio of GFP-expressing C57BL/6 (βACT-EGFP; CD45.2) and IFN-γR–deficient (CD45.2) bone marrow. Donor cells and host cells were identified by CD45 allelism; wild-type cells (CD45.2+GFP+) were distinguished from IFN-γR–deficient cells (CD45.2+GFP−) on the basis of GFP expression (Fig. 6A). When mixed with C57BL/6 cells, the bone marrow-derived IFN-γR–deficient cells were fully capable of reconstituting recipient mice, and at 6 wk postreconstitution, IFN-γR–deficient cellsdominated in the peripheral blood (40.85 ± 6.38% wild-type versus 55.91 ± 6.43% IFN-γR–deficient). Moreover, IFN-γR–deficient T lymphocytes were found at higher frequency than wild-type T cells and, notably, double-positive CDllb and Gr-1 myeloid cells were predominantly of wild-type origin (data not shown).
The chimeric mice were infected with E. muris, and the frequency and phenotype of wild-type and IFN-γR–deficient cells were analyzed within the bone marrow. Bacterial infection in the chimeric mice was similar to those in C57BL/6 mice, indicating that the chimeric mice controlled infection (Fig. 6B). We next addressed whether the donor cells were of wild-type or IFN-γR–deficient origin in the myeloid progenitor cell populations. In mock-infected mice, we observed an increased frequency of IFN-γR–deficient donor cells, relative to wild-type donor cells, within the Lin−IL-7Rα−c-KithiSca-1− (i.e., CMP) population (Fig. 6C, 6D). However, a greater proportion of the Lin−IL-7Rα−c-KithiSca-1+ cells was of wild-type origin. Postinfection, we found that the frequency of wild-type cells was greatly diminished within both the c-KithiSca-1− and c-KithiSca-1+ fractions of Lin−IL-7Rα− cells; these two populations of Lin−IL-7Rα− myeloid progenitors were instead composed almost entirely of IFN-γR–deficient cells. These data suggest that hematopoietic cells unable to signal via the IFN-γ pathway are retained as immature cells within the bone marrow, perhaps as a consequence of their inability to undergo proper differentiation. In contrast, wild-type cells, capable of receiving IFN-γ–induced signals, were not retained in the bone marrow, possibly because they differentiate and/or egress from this site. Thus, we propose that in the absence of IFN-γR signaling, myeloid cells are prevented from completing their differentiation in response to infection-induced stress.
We next examined the bone marrow and spleens of mock-infected and E. muris-infected chimeric mice to determine the proportions of wild-type and IFN-γR–deficient cells within immature and mature granulocytic populations. In mock-infected chimeric mice, IFN-γR–deficient cells predominated within the CDllbloGr-1lo bone marrow population, but postinfection wild-type cells increased in proportion (Fig. 6F). There was little change in the composition of the CDllbloGr-1hi population postinfection, and the IFN-γR–deficient cells predominated. However, within the mature granulocyte population (CDllbhiGr-1hi) in the bone marrow, we found that E. muris infection resulted in a significant increase in the frequency of wild-type cells. Moreover, the higher proportion of wild-type CDllbhiGr-1hi cells was also observed in the periphery: CDllbhiGr-1hi cells were almost entirely of wild-type origin in the chimeric mice, both prior to and after infection (Fig. 6H). There was little effect of IFN-γ signaling on lymphoid lineage chimerism in mock-infected and E. muris-infected mice (data not shown), suggesting that IFN-γ acts on myeloid, but not lymphoid, differentiation during infection. Our data indicate that IFN-γ signaling is not required for the generation of CDllbloGr-1lo cells, but is important for mediating the terminal differentiation of neutrophils.
IFN-γ–mediated transcriptional control of infection-induced myelopoiesis
Although our data suggested that IFN-γ acts directly on hematopoietic progenitor cells, we wanted to address whether the IFN-γ signaling pathway was activated in the bone marrow promyelocytic CD11bloGr-1lo cells that accumulated during infection. Therefore, CD11bloGr-1lo cells were purified by flow cytometry from mock- and day 15-infected wild-type and IFN-γR–deficient mice (>89% purity). Next, these cells were evaluated for the expression of genes known to be regulated by IFN-γ, including the IFN regulatory factors (IRFs), a family of transcription factors that are critical for various aspects of hematopoiesis and immune cell function. The expression of several irf genes was induced during infection, including irf-1, irf-3, irf-4, irf-5, irf-7, and irf-8. Compared to their expression in wild-type CD11bloGr-1lo cells, transcription of several of the irf genes was not induced in the IFN-γR–deficient cells harvested from infected mice (Fig. 7A), in particular irf-1 and irf-8. We also examined expression of a number of additional IFN-γ–regulated genes, including adenosine deaminase acting on RNA (Adar)1, Irgm1, Ifng, Ifngr1, and others (Fig. 7B). Most of these genes, known to be regulated by IFN-γ, were induced in wild-type CD11bloGr-1lo promyelocytes during infection, but not in IFN-γR–deficient cells. Thus, infection-induced IFN-γ directly enhances myelopoiesis, likely via its ability to induce the expression, in developing promyelocytes, of transcription factors known to be involved in hematopoiesis and granulopoiesis. These data also illustrate that IFN-γ is solely responsible for inducing transcription of irf-1, irf-8, Adar, Irgm1, Cxcl10, Mx1, Csf3, and Csf2rb during infection, as these genes were not induced in cells that could not signal via this pathway.
In this study, we demonstrate that an intracellular bacterial infection causes major changes in the differentiation of hematopoietic progenitor cells via direct IFN-γ signaling. We conclude that while homeostatic production of monocytes and granulocytes is not dependent upon IFN-γ signaling, infection elicits the production of both populations of myeloid cells, and that this infection-induced process is critically dependent on IFN-γ signaling. These findings reveal a proposed, but largely unrecognized, role for IFN-γ in the direct modulation of hematopoiesis during bacterial infection. Because many pathogens induce IFN-γ production, these findings have implications for understanding host defense against a wide range of microbial infections. This conclusion is supported by a report that IFN-γ induces the expansion and proliferation potential of Lin−Sca-1+c-Kit+ cells (30). Moreover, studies of hematopoiesis during malaria infection revealed the emergence of a unique population of Lin−IL-7Rα+c-Kithi progenitor cells that was found to require IFN-γ signaling (20). We propose that these observations together provide a paradigm whereby inflammatory cytokines, such as IFN-γ, act to modulate the early differentiation of hematopoietic progenitor cells, thereby enhancing the production of the mature blood cells required to combat infection.
Although our findings have demonstrated an important role for IFN-γ in vivo, they contrast with earlier studies that described a negative effect of IFN-γ on hematopoiesis. In those studies, IFN-γ inhibited in vitro colony formation of human granulocyte-macrophage progenitor cells (14, 15, 32), and even low levels of IFN-γ inhibited hematopoiesis in long-term bone marrow cultures (13). We observed that the frequency of in vitro bone marrow colony-forming cells decreased during infection in wild-type mice. Although this finding would appear to be consistent with the earlier studies that demonstrated hematopoietic suppression, we propose instead that in vitro colony formation potential decreases because IFN-γ promotes hematopoietic progenitor cell differentiation, thus depleting the bone marrow of progenitor cells. In our studies, the classically defined myeloid progenitor cells were diminished postinfection in wild-type mice but not in IFN-γR–deficient mice, and in the wild-type mice, the frequency of Lin−IL-7Rα−c-KithiSca-1+ progenitors increased. Thus, one possible explanation is that the Sca-1− cells have a greater ability to form colonies in vitro, whereas the infection-induced Lin−IL-7Rα−c-Kithi Sca-1+ population requires additional signals not provided in the standard in vitro culture conditions. Furthermore, in our studies of hematopoiesis in mixed bone marrow chimeric mice, we observed that infection caused a loss of wild-type cells within the Lin−IL-7Rα−c-Kithi Sca-1− and Sca-1+ populations; in these mice the progenitor cells were mostly derived from IFN-γR–deficient donor cells. We think that this finding is consistent with our interpretation that the wild-type cells differentiated in response to infection-induced inflammation. An alternative explanation is that these cells were mobilized and emigrated from the bone marrow. We also observed high frequencies of wild-type–derived granulocytes within the bone marrow and spleen of infected mice, suggesting that the wild-type cells that were capable of IFN-γ signaling proliferated and differentiated. Under these same conditions, differentiating promyelocytes that could not signal via IFN-γ accumulated in the bone marrow and in the spleen. Thus, we propose that IFN-γ promotes, not suppresses, hematopoietic differentiation in vivo.
Our studies reveal an important role for IFN-γ in directing the development of both granulocytes and monocytes during infection. Although our previous studies of the bone marrow granulocyte population prompted our investigation of these cells, we noted alterations to the monocyte population as well. The expansion of blood monocytes observed in C57BL/6 mice was reduced in IFN-γR–deficient mice, suggesting that IFN-γ is required for the production of infection-induced monocytes. IFN-γR–deficient mice also exhibited pronounced neutrophilia postinfection, as compared with C57BL/6 mice, suggesting that granulopoiesis does not require IFN-γ signaling. However, more careful analysis of the infection-induced granulocytes in the IFN-γR–deficient mice revealed that this population of cells was defective, as these cells were infected with E. muris. Additionally, wild-type cells that expressed the IFN-γR dominated the population of mature granulocytes in the chimeric mice. Thus, our data support a requirement for IFN-γ–dependent signaling in the terminal differentiation of both granulocytes and monocytes during ehrlichial infection.
These studies also highlight a critical role for IFN-γ in driving the transcriptional regulation of hematopoiesis during infection. IFN-γ mediates a range of signaling events within target cells. As our knowledge of IFN-γ signaling has come primarily from studies of macrophages and other differentiated cell types in vitro, how such signals are regulated in hematopoietic progenitor cells is unknown. CDllbloGr-1lo cells from infected IFN-γR–deficient mice expressed reduced amounts of irf-1 and irf-8 mRNA, relative to cells from wild-type mice. IRF-1 and IRF-8 (also known as IFN consensus sequence binding protein) are both known to play critical roles in hematopoiesis (33). IRF-1 deficiency results in increased numbers of immature granulocytic precursors (33), an observation that is similar to what we observed in infected IFN-γR–deficient mice. IRF-8 is required for normal monocyte and dendritic cell development (34, 35), which is consistent with our observation that fewer blood monocytes, but increased neutrophils, were detected postinfection in the IFN-γR–deficient mice. IRF-8 deficiency also results in myeloid hyperplasia (36) and in increased numbers of myeloid-derived suppressor cells (37); additionally, human myeloid malignancies show much lower than normal IRF-8 expression (38). IRF-8 also regulates the proliferation-inducing effects of GM-CSF by inducing the expression of neurofibromin 1, a tumor suppressor (39, 40). IRF-8 is also required for host defense: IRF-8–deficient mice are highly sensitive to Mycobacterium tuberculosis and Plasmodium chabaudi infections (41, 42). IRF-8 acts to promote phagocyte development and maturation by inducing the expression of the phagocyte oxidase proteins gp91phox and gp67phox (43). Our observation that a high frequency of granulocytes contained morulae in the spleens of IFN-γR–deficient mice is consistent with the role of IRF-8 in promoting phagocyte development during infection. Thus, the induction of irf-8 expression during ehrlichia infection likely plays an important role in regulating myelopoiesis by controlling proliferation, differentiation, and oxidative activity of phagocytes, and it is likely to be directly responsible for the defects in myelopoiesis in our infection model. Additionally, our studies show that IFN-γ–mediated signaling is critical for regulating irf-8 gene expression during infection.
IFN-γ–driven hematopoiesis can be viewed as a favorable response to infection, due to the production of pathogen-killing leukocytes. However, several studies have suggested that IFN signaling in HSCs may also have a negative effect. In this study, we have reported direct action of IFN-γ on myeloid progenitor cells, but it is likely that IFN-γ also acts on HSCs during ehrlichial infection, as several IFN-γ–inducible genes are vital to HSC maintenance (17, 18). The immunity-related GTPase Irgm1 (also known as Lrg-47) was induced in HSCs during infection and was required for the maintenance of the HSC pool (17). The IFN-γ–inducible gene Adar1 was also shown to be critical for HSC maintenance: loss of ADAR1 in hematopoietic cells led to rapid apoptosis and the induction of IFN-responsive genes (18). These studies provide strong evidence that IFN-γ acts directly on HSCs, and it was recently shown that IFN-γ signaling leads to HSC exhaustion during chronic mycobacterial infection (31). This conclusion is consistent with the observation that HSCs lacking IRF-2, a transcriptional repressor of type I IFN signaling, undergo exhaustion when chronically stimulated with IFN-α (44). Thus, although IFN-γ signaling may be an important driving force in responding to infection, this factor may also deplete stores of stem and progenitor cells within the hematopoietic compartment.
Although it is not clear how Sca-1 functions in progenitor cells, it is thought that Sca-1 can regulate different signaling pathways by modulating lipid raft composition, and it has been shown that overexpression of Sca-1 can inhibit myeloid differentiation (45). This observation is inconsistent with our hypothesis that IFN-γ drives myelopoiesis during infection. We propose that other genes, expressed during infection, modify the effects of Sca-1 signaling in vivo, as it is well known that IFN-γ signaling is context-dependent. Indeed, the observation that Lin−Sca-1+c-Kit+ cells can be induced to proliferate in response to IFN-γ (30) suggests that the outcome of IFN-γ signaling may differ in less differentiated progenitor cells. In support of this idea, it has been shown that Sca-1 is required for IFN-α–driven HSC proliferation (19).
Several reports have also revealed that TLR signaling is critical for HSC and progenitor cell differentiation (1, 3, 46–48), indicating that some pathogens interact directly with stem and progenitor cells. For example, monopoiesis was impaired in Listeria monocytogenes-infected MyD88-deficient mice (48). We observed that MyD88-deficient mice exhibited a phenotype similar to that of E. muris-infected IFN-γR–deficient mice with respect to the reduced Sca-1 upregulation on bone marrow progenitor cells (K. C. MacNamara and G. M. Winslow, unpublished observations). However, E. muris is not known to encode canonical TLR ligands (i.e., LPS and peptidoglycan), and we failed to detect any differences in hematopoiesis in infected mice deficient in TLR2, TLR4, or TLR9 (K. C. MacNamara and G. M. Winslow, unpublished observations). The TLR-associated signaling molecule MyD88 forms a physical association with the IFN-γR, and MyD88 can stabilize IFN-γ–induced gene transcripts (49). Thus, we propose that an alternative TLR-independent role for MyD88 in infection-induced hematopoiesis is via the IFN-γ pathway or in IL-1β or IL-18 signaling.
The role for IFN-γ specifically during periods of stress and infection is highlighted by the fact that under steady-state conditions the bone marrow, blood, and spleen have nearly identical frequencies of leukocytes in both IFN-γR–deficient mice and C57BL/6 mice. However, infection of IFN-γR–deficient mice resulted in enhanced frequencies and numbers of myeloid progenitor cells, as compared with C57BL/6 mice. In one model of why myelopoiesis is enhanced in the absence of IFN-γ, it was proposed that IFN-γ acts to suppress the innate immune response by acting on myeloid progenitor cells (16). This model is in conflict with our finding that in mice containing both wild-type and IFN-γR–deficient cells, most mature neutrophils were found to be of wild-type origin. Our data support the notion that IFN-γ induced during infection is required for the production of mature neutrophils. However, an alternative possibility is that IFN-γ acts to increase the lifespan of mature neutrophils, thus favoring the accumulation of wild-type cells in the spleen. We propose a model whereby IFN-γ acts directly on progenitor cells and immature cells, thereby accelerating their movement out of the pool of earlier progenitor cells. This would explain why the pool of Lin−IL-7Rα−cKithi cells is composed of IFN-γR–deficient cells in the infected chimeric mice: in the absence of IFN-γ signaling they fail to differentiate and, thus, dominate this pool.
Our data suggest that the hematological alterations observed during ehrlichiosis are due to the direct effects of inflammation on hematopoietic stem and progenitor cells. We propose that anemia and thrombocytopenia, which are hallmarks of ehrlichiosis, occur at the expense of promoting the production of granulocytes. The observation that the onset of these hematological abnormalities is delayed in IFN-γR–deficient mice, where we also see a reduced loss of classically defined myeloid progenitor cells, supports our conclusion that anemia and thrombocytopenia are a direct result of altered functional capacity of myeloid progenitor cells. The cost of producing more granulocytes may therefore be the transient loss of hemoglobin and platelet production; this outcome may be favorable to the host, as wild-type mice survive infection and IFN-γR–deficient mice are unable to control the infection.
Most infectious diseases cause inflammation and induce the expression of IFN-γ, so similar alterations in hematopoiesis likely occur in many infections. A better understanding of how infections alter hematopoiesis and progenitor cell function and capacity is critical not only for our overall understanding of immunity, but for our general knowledge of how hematopoiesis is modulated in the face of inflammation and immunologic stress.
We thank Susan Wittmer (Trudeau Institute) and Naomi Walker (University of California, Davis) for excellent technical assistance. We also acknowledge Renjie Song, in the Wadsworth Center Immunology Core Facility, and Chang-Uk Lim, in the Ordway Research Institute Flow Cytometry Facility, for excellent technical expertise.
Disclosures The authors have no financial conflicts of interest.
This work was supported by U.S. Public Health Service Grants R01AI064678 (to G.M.W.) and F32AI077242 (to K.C.M.).
Abbreviations used in this article:
- adenosine deaminase acting on RNA
- common lymphoid progenitor
- common myeloid progenitor
- enhanced GFP
- granulocyte monocyte progenitor
- hematopoietic stem cell
- human monocytic ehrlichiosis
- IFN regulatory factor
- Lin
- lineage
- megakaryocyte-erythrocyte progenitor.
- Received June 8, 2010.
- Accepted November 10, 2010.


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German Execs Risk Prison in Telecom Case
Associated Press Writer
The head of Deutsche Bank and five other former top officials of German wireless operator Mannesmann risk up to 10 years in prison if convicted of charges they arranged illegal payoffs during the company's 2000 takeover by Britain's Vodafone, prosecutors said Tuesday.
Detailing charges announced last week, prosecutors in the city of Duesseldorf alleged that former Mannesmann executives unlawfully enriched themselves during the buyout battle.
They listed multiple counts of grave breach of trust against six former Mannesmann supervisory board members or managers. No trial date has been set.
Those charged included Deutsche Bank chief executive Josef Ackermann, head of Germany's largest bank; Klaus Zwickel, head of Germany's powerful industrial union IG Metall; former Mannesmann chief executive Klaus Esser; former Mannesman supervisory board chairman Joachim Funk and two other former Mannesmann officials.
The case flowed from Esser's decision to end his opposition to Vodafone's hostile bid, followed by about 250 million marks ($137 million) in payments to Esser and other Mannesmann officials once the deal went through.
Esser conspired with Funk to authorize special payments for himself and other Mannesmann executives "in return for their support for a 'friendly' takeover of Mannesmann AG by Vodafone AirTouch PLC" during the takeover battle, prosecutor Hans-Reinhard Henke alleged.
Esser alone received about 32 million marks ($17 million) under the arrangement, which Ackermann and Zwickel approved as supervisory board members, prosecutors said.
Henke said "the payments served purely for the enrichment of the beneficiaries in connection with Mannesmann's takeover, not the company's good."
Prosecutors said the payoffs and bonuses drained up to 56.8 million euros ($61 million) from Mannesmann. That allegation underpins the breach of trust charges, which Henke said are punishable by up to 10 years in prison _ though any penalty is likely to be much lower.
Esser has repeatedly denied wrongdoing, arguing that the majority of Mannesmann shareholders favored a takeover by the time he gave up the fight. The stock swap deal was valued at $180 billion _ at the time, the largest corporate merger ever.
Ackermann, Zwickel and Funk also denied any wrongdoing. Ackermann has criticized the investigation, questioning whether Germany _ Europe's biggest economy _ should allow judges to set guidelines for corporate pay.
Zwickel expects to be cleared if the case goes to trial. In a statement issued by his union, he said he abstained in the supervisory board vote that cleared the way for the payouts.


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I took this photo using the only camera I have which is my camera phone! :'( I'm in tears. I'm missing my camera big time and badly! It's such a wrong timing and it sucks. But I really have to move on. Ugh!
Behind us playing the coolest mix songs is DJ Callum! Wasn't able to have a solo photo with him since he was running busy!
Grabbed this photo from SM Lanang Premier's album on facebook. Credits! :-)
So I was talking earlier, by 6pm he was at the atrium playing some mixes and the meet and greet sesh with his adorbs fans! :-) He's really humble and note this, VERY HANDSOME!
This guy is so adorable I could die! Hahaha! Kidding! Well, he really is adorbs wearing DJ CALLUM <3 DAVAO shirt. I think Souvenir Shop Davao gave this to him :-)If you wanna visit SM Lanang Premier which is located in J.P. Laurel Avenue, Davao City you can see this huge lettering of DAVAO in the Fountain Court. You can also watch their Waterworks (Dancing fountain) for free! See the poster below.
How was your weekend my dearly readers? Mine was so much fun since I just celebrated my 17th Birthday last March 14. Which was my camera was also damaged. But then, I had fun. To sum it up, my friends made me so happy and I wish this days wouldn't end but sadly, we have to say goodbye and face the back-to-normal days which is school. I am so ready for summer, in case you don't know. The heat we are experiencing now is insane! Well, talking back about the Spectrum Party, it was so nice seeing DJ Callum play and he's really handsome. Very, rather! Anyway, if you know me and took a photo of me/us last night please do tag us on Twitter @eishellecellona or on Facebook: Eishelle Cellona. Thank you! Kisses! :*
I hope you enjoyed my post as much as I did even though the quality of the photos aren't that great but I really promise you that as soon as I get my DSLR back, I'm gonna flood you with so much of the great photos! Enjoy your week ahead, sunnies!


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Stream of Consciousness
A steady flow of thoughts about advertisingand marketing from True Creek.
Posted by truecreek on November 5, 2012 under The Work | Be the First to Comment
Couple of layouts from a Dust Wall project at Dulles International Airport.
Tags: advertising agency in Northern Virginia, consumer-marketing
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© 2019 Stream of Consciousness
Balance of Blue
by Brandon Cox.


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Almost all of us have studied history in school. Some, I'm sure, enjoyed it more than others, and some of you are currently in education trying to remember how many wives Henry VIII had. But sift through your history textbooks, how much of an emphasis is put on Welsh history? Not enough you might say? A fair amount maybe? Perhaps even too much?! Well its place on the curriculum may very well change after the education minister, Leighton Andrews, ordered a review into the way it's taught in school. A little over a month on it's time to look closer at what place Welsh history should be given.
Where's the Welsh History?
The major exam body in Wales, the WJEC, suggests that of the eighteen optional GCSE units in the its syllabus, the five units that deal with Germany or the USA account for 72% of registrations in 2012/13. On the other hand the six units that deal with 'Wales and England' or Wales only, account for less than 15% of registrations.
Last year around 10,000 pupils in Wales sat the WJEC GCSE history paper. All these candidates would have sat at least one unit, 25%, on aspects of the history of Wales and England. The WJEC do offer one option specifically devoted to Welsh history in the twentieth century. Only 23 candidates sat the paper, all from one centre. Although these figures only account for the WJEC GCSE history syllabus over one year, they make for interesting reading.
At the same time over the last decade there has been a significant growth in interest in the history of Wales, culminating in the BBC's The Story of Wales with Huw Edwards. Online educational resources to support the study of Welsh history have also continued to grow. The Welsh Government has invested heavily in digitization projects through the National Library of Wales and Cymal for instance. And on November 13th The Story of Wales Director announced via Twitter that they're versioning the series for classroom use. What better time to consider the place of Welsh history within the curriculum?
The Review
The review group, led by Dr Elin Jones, will explore and advise on the future development of the history curriculum and report back in July 2013. The 13 member group was handpicked by Dr Jones and is made up of key individuals with experience in Welsh history and its teaching. The other members of the panel are:
- Prof Angela John, Aberystwyth University;
- Dr Sian Rhiannon Williams, Cardiff Metropolitan University;
- Sion Jones, Ysgol Syr Hugh Owen;
- Dr Hugh Griffiths, Ysgol Bro Myrddin;
- Paul Nolan, History advisor;
- Nia Williams, Education Coordinator, National Museum and Galleries of Wales;
- Frank Olding, Blaenau Gwent Heritage Officer;
- Dr Stephanie Ward, Cardiff University;
- David Stacey, Olchfa Comprehensive School;
- Dr Martin Johnes, UC Swansea;
- William Rogers, Queen Street School, Blaenau Gwent;
- Nicola Thomas, Cornist Park School
'Lack of Resources'
So is there a lack of Welsh history in the current curriculum? Andrea May, history teacher and Assistant Head at Brynteg Comprehensive School in Bridgend said: "If you actually look at the kind of materials that you get [on Welsh history] they're not engaging for the students. You've got things like NGFL which I think will probably grow even more, but that relies in many respects on teachers feeding into it as well. And if it isn't being taught you're not going to get that share in the resources."
She added that as children went through the year groups there is a decline in how much Welsh history is studied. In Brynteg their A-level course offers only one aspect of Welsh history, and that as part of a unit on Britain.
Is the teaching of Welsh history being overlooked then? Dean Powell, a local historian from Llantrisant believes it is, especially at comprehensive level. Drawing from his own experiences he said: "By the time I went to Ysgol Gyfun Llanhari [a Welsh medium comprehensive] the curriculum was far far wider and then all of a sudden you're learning everything from the French to the Russian revolution and the rise of Nazism… but actually what you ended up losing was the Welsh history aspect."
A School Trip
As Dean explains, today's children are incredibly lucky to have so many resources at their disposable. To see how history is brought to life I joined Year 3 from Ysgol Gynradd Bro Eirwg in Cardiff on their school trip to St Fagans National History Museum and spoke to their teacher Mrs Bethan Elis.
Ethnic History
When the review was announced Oliver Hides on his Morning Call show on BBC Radio Wales spoke to Val Williams, a social historian and former teacher who said:
"Take Cardiff for instance; there are kids living in Cardiff whose families came from Somalia – what brought them here? What kept them here? How do they see the past of the city around them?" (Read the BBC story here)
One man who agrees with Val is Ibrahim Harbi, the National Co-ordinator of the Somali Integration Society who had this to say:
Whether the curriculum changes or not, the opportunities for school children are endless. This map shows the number of museums and places of historical interest in South East Wales. If they don't inspire our kids then what will?!


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I bought a huge bag of sweet glutinous rice to make Andy Ricker's Sticky Rice with Mango and Salty-Sweet Coconut Cream (https://food52.com/blog...), which was incredible. What should I do with the rest of this glutinous rice? Are there any ways to use it that aren't just coconut sticky rice (not that there's anything wrong with that...)
I often use glutinous rice when making rice porridge (jook). It makes a creamier porridge.
This is yummy: http://thewoksoflife.com/2015/04/sticky-rice-with-chinese-sausage/
Another version:
Just an FYI, TheWoksOfLife.com is one of my favorite blogs.
Sweet rice is stickier than short grain rice but is very tasty. You can mix it with long grain rice like jasmine or Basmati rice and use it like a short grain rice in a recipe. I usually one cup of sweet rice to two cups of long grain rice.
Indonesian Lemper (easier version): http://dailycookingquest.com/by-cuisine/indonesian/lemper-ayam-steamed-glutinous-rice-with-spiced-chicken-floss
could you make sweet white rice flour (via food processor)? have seen that as an ingredient in gluten free baking


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A powerful logo opener with cinematic synth elements, deep percussion, distorted beats and strings.
You get three separate edits with this purchase:
- Cinematic Power Logo 01 – Full (duration 0:19)
- Cinematic Power Logo 01 – Beat Start (duration 0:11 – starts at 0:08 in the preview)
- Cinematic Power Logo 01 – End Impact (duration 0:06 – starts at 0:13 in the preview)
If you like this track, do rate or leave a comment – I'd love to hear your thoughts.
Also be sure to check out my other intros and openers here.
Videohive Authors, you're very welcome to use the watermarked version of this track in your projects if you link to this item – and do let me know about it, so I can feature your work.


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Featuring data transfer speeds of up to 30MB/s, Transcend's Class 10 SDXC/SDHC cards provide an impressive level of performance for high-megapixel digital cameras and Full-HD camcorders. With impressive write speed, these cards provide enhanced camera response for high-speed consecutive shooting as well as smooth Full HD video recording.
View or download the product sheet for detailed product and ordering information.
Find drivers, manuals, FAQs, or software updates for your product.
Check the compatibility of your Transcend product with your computer system or electronic devices.


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If you're studying Stone Age to Iron Age Britain, what better way to learn than to be out in the woods living like Stone Age people or visit a real Iron Age hillfort?
Children will experience 500 and 5000 BC when they come to Wendover Woods, and get to compare the Stone Age with the Iron Age.
You will spend half the day in the Mesolithic, around 5000 BC. In the woods, you will learn how to build shelters, make fire and cordage and make flint tools.
In the other half of the day you will scoot forward in time to 500 BC, the Iron Age, take a tour of a Boddington hillfort and then come back to the homestead to do some chores, like grinding grain on a quern, making butter and braiding wool. You'll finish the day with recreating an attack on the hillfort with slingshot (grapes!)
- One class, full day £250
- Two or three classes, full day £450
If you'd like to book, use our contact form to get in touch.


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Dec. 7 vs. Nashville Predators at Verizon Center
Time: 7:00 p.m.
Radio: Capitals Radio Network
Nashville Predators (13-13-3)
Washington Capitals (14-12-2)
Washington concludes a quick two-game homestand on Saturday night when it hosts the Nashville Predators at Verizon Center. For the Capitals, the game is the front end of the team's third set of back-to-back games in as many weekends and its fourth set of back-to-backs in the last five weekends.
Although Washington is tied for second place in the NHL's Metropolitan Division and the Predators are tied for last in the Central Division, the Caps are only one point ahead of the Preds overall.
The Caps' modest two-game winning run came to a halt on Tuesday when they absorbed a 4-1 loss at the hands of Metro foe Carolina. Although the Caps were happy with the way they played for the first 30 minutes of the game, they trailed 1-0 at the midpoint of the contest. Carolina blew it open with three goals in less than four minutes just past the midpoint of the second, enabling the Hurricanes to take a 4-0 lead into the third period.
Washington spoiled Justin Peters' shutout bid early in the third when Caps defenseman Mike Green scored on a power play, his first goal of the season.
When the Capitals returned from a tough set of weekend back-to-back games on the road in Phoenix and Colorado just over a month ago, the team looked ahead to a favorable stretch of scheduling that had the Caps playing 23 straight games in the Eastern time zone and going 47 nights without being on the road for consecutive nights.
"It's big for us," says Caps right wing Troy Brouwer of the team's schedule. "You saw that first month of the season where we spent a lot of time in the central and the western time zones, we struggled a little bit with the time changes. So being in our own time zone, being around our families and being home a lot, it's an advantage for us. And we've got to take advantage of it.
"I know guys are excited to be home and have a lot of home games around the holidays. It's always tough when that time of year comes around and you're on the road a lot. We've been fortunate in our scheduling, and I know [Caps general manager] George [McPhee] works hard on that to make sure that we have as favorable as possible of a schedule."
It looked as though the Caps were poised and primed to take advantage of that stretch of games, which comprises more than a quarter of the season. Washington won three straight games, putting itself in position to take over first place when the Penguins visited on Nov. 20. But Pittsburgh whitewashed the Caps 4-0, setting them off on their current 2-4-1 tailspin.
Although the Capitals are 9-5-2 since the start of November, they go into the Nashville game with just one regulation win in their last 13 games. That covers a span of 31 days.
Goaltender Braden Holtby didn't make it through the Carolina game on Tuesday; he was pulled in favor of rookie Philipp Grubauer after the second period. Holtby surrendered four goals on 23 shots. A hallmark of Holtby's résumé over his brief NHL career is the ability to bounce back after bad losses and/or difficult outings.
In 20 starts that have followed games in which Holtby was either pulled or allowed four or more goals, he is 14-4-1 with two shutouts, a 2.11 GAA and a .930 save pct.
The only time this season that Holtby went consecutive starts without helping the Capitals to earn at least a point was Oct. 5-10 when he dropped consecutive regulation decisions to Dallas (2-1) and Carolina (3-2), respectively.
Caps goalie Michal Neuvirth has been sidelined since Nov. 29 when he stepped on a puck as he took the ice for warm-ups before a scheduled start against the Montreal Canadiens. On Friday, Neuvirth made it through his first full practice since sustaining the injury.
"It's getting better every day," says Neuvirth. "I'm happy I made it through a first practice. It's a good first step. I've just got to keep moving forward and getting better."
Neuvirth doesn't believe he'll be ready for game action this weekend. With a second game in as many nights on Sunday against the Rangers in New York, the Caps could turn to Grubauer to start that second game. Washington bench boss Adam Oates says the decision will likely be based on how things go on Saturday against the Predators.
As mentioned, Washington is tied for second place in the Metropolitan with the New York Rangers. But the Caps have been idle since that loss to the Hurricanes on Tuesday. Depending on the outcome of NHL games between now and puck drop against the Predators on Saturday night, Washington could have the company of two other teams in that second-place tie with the Rangers, and the whole group of teams could fall beneath Carolina in the standings.
New Jersey (28 points) hosts Detroit on Friday night, and Philadelphia (28 points) visits Dallas on Saturday afternoon. Carolina (29 points) could move into sole possession of second if it is able to author a win on home ice against San Jose on Friday. Seventh-place Columbus (25 points) plays host to Minnesota on Friday. If the Jackets win, they'll still be in seventh. But they'd be only three points shy of Washington.
This weekend's set of back-to-backs looms crucial for the Capitals.
Nashville comes to town having lost four straight games (0-3-1). The Predators scored a combined total of five goals in those four losses, the most recent of which was a 5-2 setback at the hands of the Carolina Hurricanes on Thursday night. The slide encompassed a four-game homestand in which Nashville managed just one of a possible eight points.
With an average of 2.24 goals per game, the Predators are tied for 24th in the NHL in that category and Nashville's .71 ratio of five-on-five goals for and against is 28th in the league.
While Nashville is traditionally a defensively stingy team, they suffered for a while from the loss of goaltender Pekka Rinne, sidelined since October with a hip infection. The Preds hope to have Rinne back practicing later this month.
In Rinne's absence, the Predators initially turned to first-year journeyman Carter Hutton. Hutton had a few good games early in the season, but then endured a stretch of six straight appearances in which he surrendered three or more goals.
Enter Marek Mazanec. The Preds' sixth-round choice (179th overall) in the 2012 NHL Draft, Mazanec was recalled from AHL Milwaukee on Nov. 5. Less than a month later, he was named the NHL's rookie of the month for November. In his first month in the circuit, Mazanec posted a 5-4-1 record to go along with two shutouts, a 2.00 GAA and a .932 save pct. in 11 appearances (10 starts).
Mazanec has dropped his first two decisions of December.
"They're one of the teams that really stick together," says Caps forward Eric Fehr of the Predators. "They play as a five-man unit and it makes it difficult; it doesn't matter who they have in the lineup there. They're tough. If you beat one guy, you've got to beat two and so on. They play a good game and obviously we're going to have to try to work around it."
The Predators got some good news on Friday when franchise defenseman Shea Weber practiced with the team for the first time since taking a puck to the eye in a Nov. 28 game against the Edmonton Oilers. The Preds went on to lose the game against the Oilers, and they went 0-2-1 in the three games Weber missed. Weber is tied for the league lead in goals among defensemen, and the Preds are 7-0-0 when he scores this season.
Saturday's game marks the return of ex-Caps forward and fan favorite Matt Hendricks. Last July, Hendricks signed a four-year contract with Nashville as an unrestricted free agent. He spent the previous three seasons with the Capitals, totaling 18 goals, 42 points and 278 penalty minutes in 203 games with Washington.


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Press release
CAP Reform: Dual use will continue to be allowed in 2015
Farmers and landowners in England with Environmental Stewardship agreements will be able to maintain their existing dual use arrangements in 2015.
Farmers and landowners in England with Environmental Stewardship agreements will be able to maintain their existing dual use arrangements in 2015. The Rural Payments Agency's (RPA) External Relations Director Arik Dondi, said: "This decision, announced this week, covers all existing Environmental Stewardship agreements, including those remaining to be signed up this year, and means the continuation of two different beneficiaries receiving support under two separate CAP schemes for the same piece of land at the same time, provided the different parties can each meet the rules of the different schemes.
"We are committed to making information available to help people understand how the new CAP will affect them as soon as decisions have been taken and I am sure our Basic Payment Scheme customers will welcome this latest detail.
"We are determined to implement the CAP in a way that is as simple, affordable, and effective as possible and to ensure that the countdown to the new scheme is as smooth as possible for farmers and landowners."
Tenants or landlords whose land will be in dual use in 2015 should be aware that with the introduction of greening requirements, where land is let out under a tenancy agreement (Agricultural Holdings Act or Farm Business Tenancy), the tenant will need to include that land when working out their greening requirement. The landlord would not include that land as part of their greening requirement.
It is important that anyone planning to claim a Basic Payment Scheme (BPS) payment next year for land that someone else is claiming for under another scheme has a written record they have the land at their disposal and that they are eligible for the scheme. They will also need to read the leaflet called Extra guidance on dual use of land.
Further information about how dual use will work for the new environmental land management scheme will be released at a later date.
Up-to-date information about CAP Reform is available through the GOV.UK website at www.gov.uk/cap-reform.


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Western Montana 3/4 Mile Riverfront
81 Brockway Lane : Superior, MT 59872
Mineral County, Montana
Land Description
Western Montana Clark Fork Riverfront land with 3/4 mile river frontage, Beautiful riverside level benches building sites overlooking the river and mountains. Great area for fishing, hunting, rafting. Beautiful 95.61 acres on the Clark Fork River of Western Montana with approximately 3/4 mile of river frontage. Beautiful riverside building sites on level benches well above the floodplain with stunning river and mountain views. The land is a nice mix of meadows, trees and river frontage. Great location just a few miles east of Superior with grocery and hardware store even a hospital and paved airstrip. Less than an hour to the Missoula Airport and all the amenities including the University of Montana and Costco. Excellent access with power adjacent to property. Area of excellent trout fishing and rafting. Wildlife include Waterfowl, Eagles, Osprey, Deer, Bear and Elk have all been known to frequent the property. Contact your Realtor or John Horton for a tour of this very nice riverfront land.


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- All Jewelry
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Popular & New
This item is available for backorder and will ship within 2 to 8 weeks. Read more
This item is available for pre-order and will ship within 2 to 8 weeks. Read more
"I was inspired by the beautiful nature that surrounds us, especially the extravagant unusual shape of seahorses," confides Felix Carballo about this ocarina vessel flute. The Nicaraguan artisan crafts the ceramic ocarina by hand detailing the seahorse with a bicycle spike and eyes with a fine paint brush. Carballo prepares the different colors by pulverizing stones and using a brush to splash the color, successfully achieving an aged effect. This ocarina features two tonal perforations on the front, two on the back, and the mouth piece is under the crown.
Ocarinas are one of the oldest musical instruments on the planet. Originally designed like a vessel with up to 13 perforations, ocarinas are now crafted in a variety of styles.
"I never went to school, but I learned to read and write because I wanted to. I even developed an acceptable calligraphy, and that makes me very happy."


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This kid I've known for five or so years, but literally haven't talked to since 7th grade, just asked me on a date a few days ago. It will be both of our first dates. I used to like him, a long time ago back in 7th grade when we were friends, and then he found out. He stopped talking to me. I thought he hated me because he has literally never talked to me up until now, three years later. He talks to a lot of girls, and is good friends with a lot of girls, excluding me, so I think it's pretty peculiar that he randomly asked me out, of all people. I dont even consider us friends. not to mention this date is a big popular formal dance, very similar to prom, and he did the whole cute prom-posal and everything. A few of my friends said he had been asking them beforehand if I have a boyfriend or not, and told them he wanted to ask me to this dance and that he thought I was really nice and kind. Its super awkward that he's been saying all these things since we never talk, unless he actually has a crush on me. But I need help deciding. Maybe he just wants to be friends. We texted about our plans a little and he did say he didn't want me to be nervous and that it will be a good opportunity to make new friends. I dont know what to think and how to act around him so please give your opinions!
Most Helpful Girl
I say give him a chance! Unless you plan on asking someone else to the dance. It doesn't mean you have to be in a relationship with him.


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19 Jan 2018
Brazil Simplifies Import Procedures for Goods Subject to Sanitary Oversight
As part of a strategy to increasingly focus its enforcement efforts on higher-risk products, the Brazilian National Health Agency (ANVISA) has simplified in various ways the procedures to import goods subject to sanitary oversight. Among other changes, the import licencing process has been simplified and importers will no longer be required to submit certain documentation that was only available after their shipments had arrived in Brazil, thereby lowering storage costs. Some of the requirements that have been eliminated include the linkage of tariff lines to specific procedures, a provision that required signature authentication and recognition, a provision that required import declarations to include certain data and registration information, the batch declaration requirement, and certain certification and analysis requirements for food imports.


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Please join us in praying for the lost youth in our area and for the mission of Youth For Christ.
Current Prayer Requests
Please join us in praying for:
- Our staff as they build relationships with lost youth in the community
- New adult volunteers to expand our ministry
- Positive working relationships with state officials and local schools
- The beginning of new area ministries
Join our Prayer Support Team
"And this is the confidence that we have toward Him, that if we ask anything according to His will He hears us." 1 John 5:14 Many dedicated volunteers serve by regularly praying for our work. Join join our prayer team sign up below. Team members receive regular emails from the staff members, detailing their prayer needs.


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very good content
Navodaya Vidyalaya Samiti P.G.T. Exam. 2014 History
- Suresh
- Posted on:
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Solved Paper
Navodaya Vidyalaya Samiti P.G.T. Exam. 2014
(Held On 1-6-2014)
►Options given in 'Bold' are the answers.
1. Which of the following is not one of the Harappan sites related to the maritime
activates of the Harappan people?
a) Lothal
b) Balakot
c) Sokta kob
d) Desalpur
2. Which of the following findings helps us most to understand the religious and social
life of the Indus Valley age?
a) Burial grounds
b) Terracotta figures and seals
c) Furniture and pottery
d) Remains of houses and other buildings
3. Which of the following was not a religious belief of the Harappan people?
a) Belief in evil spirits and life hereafter
b) Cult and fire
c) Adoration of trees and animals
d) Erection of large prayer houses
4. Evidence of ploughing the land is found from?
a) Rangpur
b) Harappa
c) Kalibangan
d) Roper
5. Which one of the following is not true about Janapada?
a) Its literal meaning is the place where people place their feet
b) A Janapada was essentially an agricultural settlement
c) The Janapadas were often named after the dominant Kshatruya clans
d) The establishment of Janapada show that trade had become the
most important factor in the contemporary life
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