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Dokka, pioneering Louisiana scientist, dies
NEW ORLEANS A Louisiana State University geologist who shed light on the deep underground forces that contribute to the state's rapidly vanishing coastal wetlands has died.
Roy Dokka died Monday, LSU officials confirmed Wednesday. An exact cause of death was not immediately released. He was 59.
Dokka gained prominence by questioning the science underpinning much of Louisiana's massive and urgent coastal restoration plans. He complained that large-scale plans were moving forward even though many of the processes causing devastating land loss and subsidence in coastal Louisiana were, in his view, not acknowledged.
Coastal Louisiana, the home of New Orleans, is at the center of a vigorous scientific and socio-political debate about rising sea levels, flooding and the future of one of the nation's most important port and oil regions. Dokka's work was considered important in that debate. Since the 1930s, Louisiana has lost about 2,100 square miles of coast due to a host of problems, from levee building to deforestation.
To this debate, Dokka added his studies of the deep geologic forces at play under the soft delta muds of Louisiana. He argued that deep-seated faulting and the sheer load of sediment coming down the Mississippi River had caused Louisiana to slowly sink into the Gulf of Mexico. His theories posed a problem for Louisiana because he foresaw that the sinking would continue indefinitely.
Virginia R. Van Sickle-Burkett, a climate change scientist at the U.S. Geological Survey who specializes in the Louisiana delta, called Dokka "a prolific and highly regarded scientist in his field."
"Roy kept us all honest," she said. "He postulated things that other scientists had not considered. He was a pioneer."
But his message wasn't exactly pleasing to hear for folks in Louisiana.
"He was very pessimistic about the future of south Louisiana," said Len Bahr, a coastal scientist who had advised governors. "The bottom-line is that he says that levees are the only long-term solution, and they would not be a solution. I think he had a big part of the picture, but not the whole picture."
Bahr added: "There has not been enough emphasis on the underlying geophysics."
There is lively debate among scientists about whether coastal Louisiana, made up of the fragile delta of the Mississippi River, can be restored by artificial means, such as diverting rivers and sediment to restore marsh and barrier islands that have been eroding at alarming rates.
After Hurricane Katrina struck in 2005, Dokka worked closely with federal officials to develop a network of state-of-the-art benchmarks to accurately measure the height of structures such as bridges, levees and roads using global positioning system technology. By the time of his death, he had established a comprehensive network to track land changes. He was the director of the Center for Geoinformatics and the Louisiana Spatial Reference Center at LSU.
"The GPS stations are one the most important technological improvements since Katrina," said Windell Curole, a manager who runs a system of levees that Dokka studied and measured south of New Orleans. "In Louisiana, elevation is everything. Before (Dokka) it would take months and hundreds of thousands of dollars to get your elevation."
Tim Osborn, the regional manager for the Office of Coast Survey at the National Oceanic and Atmospheric Administration, said Dokka "was able to paint the big picture why our landscape was changing" and uncovered numerous inaccuracies in elevation across the coast.
Van Sickle-Burkett called that work "a game changer" for Louisiana.
"He showed dramatic rates of subsidence of roads and levees," she said. "Had we known some of that 20 years ago, there might have been a different layout for evacuation routes. The rates he came up with were stunning."


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Dale A. LarmieMay 14,2013Dale A. Larmie
STOCKBRIDGE A memorial gathering and potluck for Dale Larmie, 59, who died March 12, 2013, will be held from 2 to 4 p.m. Saturday, May 18, 2013, in the field at the junction of Routes 100 and 107 in Stockbridge. (BYOB and no dogs, please.)MORE IN ObituariesBeverly J. Barbour
SPRINGFIELD Beverly J. Full StoryDale W. Stafford
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text: I am running Vista MCE on my machine and have setup to output standard analog speaker to connect directly to TV input? I am using Nvidia Purevideo and set the DVD playback to S/PDIF audio outputs but it was not working. I have to go back to Vista MCE audio setting to set it S/PDIF audio outputs and then it started working. The issue is that I dont want to turn on the AMP all the time for the standard TV playing. It will not like Vista MCE allow audio to output to multiple output. How can I only setup Vista MCE to send DVD playback to S/PDIF audio outputs? Thanks for any help.


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The latest TV spot for Man of Steel continues the trend of focusing on one character, this time it's Kevin Costner that offers up some words of wisdom for his adopted Kryptonian son, Clark (Henry Cavill), as alien ships blast the Earth with blue lazer beams (that's new) and Superman gets all ROARRRR as he zips around the atmosphere and into Lois Lane's (Amy Adams) arms.
This is probably the most footage I have watched of this film yet and I like the look and the tonal vibe I'm getting and I really am hoping this thing delivers. With Christopher Nolan behind the scenes it certainly gives me a greater level of comfort.
Give it a watch below.
A young boy learns that he has extraordinary powers and is not of this Earth. As a young man, he journeys to discover where he came from and what he was sent here to do. But the hero in him must emerge if he is to save the world from annihilation and become the symbol of hope for all mankind.
Henry Cavill plays Superman in director Zack Snyder's "Man of Steel." The film also stars Amy Adams as Daily Planet journalist Lois Lane, and Laurence Fishburne as her editor-in-chief, Perry White. Starring as Clark Kent's adoptive parents, Martha and Jonathan Kent, are Diane Lane and Kevin Costner.
Squaring off against the superhero are two other surviving Kryptonians, the villainous General Zod, played by Michael Shannon, and Faora, Zod's evil partner, played by Antje Traue. Also from Superman's native Krypton are Lara Lor-Van, Superman's mother, played by Ayelet Zurer, and Superman's father, Jor-El, portrayed by Russell Crowe. Rounding out the cast are Harry Lennix as U.S. military man General Swanwick, as well as Christopher Meloni as Colonel Hardy.


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We can cater to large and small projects globally. Our speciality is providing Eco Architectural Products to suit any landscaping and building project.
Our aim is to design and manufacture a uniquely Australian product, that is created locally with passion and care, and is passed from our family to yours. At Kyoto Design we look forward to assisting you in making your dreams become reality.


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08:00 AM, Friday, May 27
PETTUS – Michael Enriquez has waited a decade and a half to be a head coach. He's poured his blood, sweat and tears into helping build winning programs throughout the South Texas area. And now, after 16 years as an assistant coach at seven differe...
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Trojan diamondmen rally on the road
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text: The Fabled Lands project was great fun, and it launched us into the work we've done since, including our own gamebook Arcana Agency and our upcoming Kickstarter for The Way of the Tiger. However, it by itself was not a financial success. Thus we're taking the lessons learned (and personal connections made) and moving on. Sorry to disappoint you, but also thank you for your interest!


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noel_of_spike - and the fahbulous sueworld2003 who is now modding it and doing a great job - thanks Sue.I hammered a few nails into my broken brain to steady it a bit (screws always go loose so quickly...) and came up with this for my offering for the wonderful
Happy Holidays, everyone!!!
Happy Holidays, everyone!!!
- Current Location:down in the dumps
- Current Mood:don't come to me for any fucking Christmas joy - spread my share amongst yourselves
- Current Music:keeping the Leonard Cohen locked up
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Army of TWO: The Devil's Cartel Coming From Visceral Games in 2013
Written Thursday, August 02, 2012 By Richard WalkerView author's profile
EA has today announced the third Army of TWO game, titled Army of TWO: The Devil's Cartel. It'll "reset" the series with Dead Space developer Visceral Games taking the reins for the third instalment in the franchise that will boast the same 2-player co-op gameplay with an added mature and grittier edge. This time it'll also be powered by DICE's Frostbite 2 Engine.
You'll play as T.W.O. (Tactical Worldwide Operations) operatives Alpha and Bravo who find themselves on the streets of Mexico, slap-bang in the middle of a drug war on an assignment mired in deceit and corruption. And as you'd expect, you'll need to work together to bring the vicious drug cartels down.
"We wanted to refresh the Army of TWO franchise for a new generation of action gamers, and at the same time, stay true to the core concept that really made the first two games fan favourites," said Julian Beak, Executive Producer at Visceral Games. "With meaningful, seamless co-op gameplay at its core and explosive new tag-team features, gamers will have to work together tactically and strategically as they push through a city dominated by one of Mexico's deadliest drug cartels."
As well as supporting online co-op, Army of TWO: The Devil's Cartel will also enable you to play in local 2-player split-screen, and you'll be able to customise and upgrade your character to distinguish them from your friend's custom operative. There'll be a deep customisation system with more options, enabling you to upgrade weapons and customise your distinctive mask.
Army of TWO: The Devil's Cartel will be releasing in March 2013.


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I have been diagnosed of GERD. I get a lot of acid reflux and if feels there is a burp which is stuck inside and if i try harder to burp, i may vomit. One doctor prescribed Rabeprazole while another gave famotidine. can I take both of them; if yes, what time of day should i take each?
Can Rabeprazole and famotidine both be taken together for GERD?
- Posted:
- 26 Aug 2009 by adi_agg
- Topics:
- famotidine, rabeprazole, gerd
Answers (1)
5 Sep 2009
You should only take both tablets if the same doctor has prescribed them both to you. Both drugs are used to treat GERD but they work in different ways. Do not take them both.
See http://www.drugs.com/enc/gastroesophageal-reflux-disease.html
Further Information
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Is there a difference between Gastroesophageal reflux and ulcer?
1 answer • 27 Apr 2010
What is the difference between Novo-Rabeprazole EC; Teva-Rabeprazole EC and PMS-Rabeprazole EC?
I have been on Rabeprazole EC for a few years for GERD and was wondering why lately the drugstore has changed it from Novo to Teva and now PMS.
1 answer • 20 Oct 2010
Can a patient take Famotidine and omeprazole for reflux and heartburn or is it too much?
This person is taking famotidine 20mg plus omerprazole 20mg is it not applied for the same thing, heartburn or acid reflux
3 answers • 22 Aug 2011
Will famotidine work for acid reflux?
Famotidine 20 mg
3 answers • 30 Sep 2013
Can I use Famotidine 20mg for acid reflux every night forever ?!?
Is there any special side effect for a long period time usage ? because when I use it every night I have no any reflux and when I stop it I'll ...
3 answers • 4 Dec 2013


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TEHRAN: Iran on Wednesday looked to expelling the British ambassador in an angry retaliation for fresh Western sanctions over its nuclear programme, which has been slammed by both China and Russia.
Iranian lawmakers, some crying "Death to Britain", adopted an emergency bill to be voted on Sunday that would downgrade diplomatic relations to the level of charge d'affaires if passed, the legislature's website said.
The bill also said parliament could take action "on other countries that behave in a manner similar to that of Britain," according to the Fars and Mehr news agencies.
Britain this week, in coordination with the United States and Canada, announced the new sanctions on Iran. They cited as justification in a report by the UN atomic energy watchdog this month suggesting Tehran was researching nuclear weapons.
Britain said it was "ceasing all contact" between its financial system and that of Iran.
The United States and Canada said they would also clamp down on the sector, including on Iran's central bank.
France has called for a freeze on Iranian central bank assets and an embargo on oil exports.
Britain, Canada and France have embassies in Tehran.
Canada's is headed only by a charge d'affaires; the other two by ambassadors.
The United States does not have a diplomatic mission, having closed its embassy after Islamic students took its diplomats hostage in 1979 following Iran's revolution.
US interests are handled by the Swiss embassy.
China on Wednesday criticised the Western sanctions, saying they would "exacerbate" the stand-off over Iran's nuclear activities.
"We believe pressuring and sanctions cannot fundamentally solve the Iranian nuclear issue. On the contrary, they will complicate and exacerbate the issue and intensify confrontation," said foreign ministry spokesman Liu Weimin.
Russia on Tuesday blasted the sanctions as "unacceptable and against international law."
China and Russia have used their weight on the UN Security Council to block any possibility of the sanctions being more broadly imposed through a UN resolution.
Iran is already subject to four sets of UN sanctions.
It has rejected the UN atomic energy agency's report, and insists its nuclear programme is exclusively for peaceful, civilian purposes.
The latest sanctions put more pressure on Iran's financial sector, with the US and Britain invoking anti-terrorist laws to target the central bank and other financial institutions.
They aim to make it more difficult for Iran to be paid for its oil exports, and put pressure on Iran's currency.
They stop short, however, of hitting the central bank with more draconian measures, which Western officials and analysts feared could cause a spike in oil prices, worsening the global economic downturn and providing Iran with a revenue windfall.
Iran's representative in OPEC, Mohammad Ali Khatibi, told ISNA news agency his country could "adopt special stances" in terms of using its vast oil exports as a political tool if "emergencies and special situations demand."
He stressed, though, that Iran was not at this time changing its approach in the global oil market.
Pressure on Iran looked likely to be raised a notch on December 1, when EU foreign ministers were expected to announce additional sanctions on some 200 Iranian firms and individuals, according to diplomats.
US President Barack Obama said in a statement Monday as the latest sanctions were unveiled: "As long as Iran continues down this dangerous path, the United States will continue to find ways, both in concert with our partners and through our own actions, to isolate and increase the pressure upon the Iranian regime."


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Get ready for more hot young gym dudes tricked out in athletic wear in Get Your Ass In Gear! Pt. 2. We've stripped away all distractions to focus on the fetish of chiseled hung hunks in jockstraps, shoulder pads, football helmets, knee pads, and protective cups getting it on like pros. These guys swallow hot cocks and fuck big bubble-butts like they're going for the gold! Put on your favorite singlet, jockstrap, or whatever it takes to get you off and Get Your Ass In Gear! Sample Scene
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This video is 70 megs - a few minutes to download to your computer...but worth every second !
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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Molecular and Functional Characterizations of Gastrula Organizer Cells Derived from Human Embryonic Stem Cells†‡§
Version of Record online: 5 APR 2011
DOI: 10.1002/stem.621
Copyright © 2011 AlphaMed Press
Volume 29, Issue 4, pages 600–608, April 2011
Additional Information
How to Cite
Sharon, N., Mor, I., Golan-lev, T., Fainsod, A. and Benvenisty, N. (2011), Molecular and Functional Characterizations of Gastrula Organizer Cells Derived from Human Embryonic Stem Cells. STEM CELLS, 29: 600–608. doi: 10.1002/stem.621
- †
Disclosure of potential conflicts of interest is found at the end of this article.
- ‡
Author contributions: N.S.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing; I.M.: collection and/or assembly of data; T.G.: collection and/or assembly of data; A.F.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; N.B.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript.
- §
First published online in STEM CELLSEXPRESS February 15, 2011.
Publication History
- Issue online: 5 APR 2011
- Version of Record online: 5 APR 2011
- Accepted manuscript online: 15 FEB 2011 04:04PM EST
- Manuscript Accepted: 4 FEB 2011
- Manuscript Received: 27 SEP 2010
- Human embryonic stem cells;
- Human development;
- Gastrulation;
- Organizer
- Top of page
- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
The Spemann-Mangold organizer is the structure that provides the signals, which initiate pattern formation in the developing vertebrate embryo, affecting the main body axes. Very little is known about axial induction in the gastrulating human embryo, as research is hindered by obvious ethical restrictions. Human embryonic stem cells (hESCs) are pluripotent cells derived from the pregastrula embryo that can differentiate in culture following a program similar to normal embryonic development but without pattern formation. Here, we show that in hESC-derived embryoid bodies, we can induce differentiation of cells that harbor markers and characteristics of the gastrula-organizer. Moreover, genetic labeling of these cells enabled their purification, and the discovery of a comprehensive set of their secreted proteins, cell surface receptors, and nuclear factors characteristic of the organizer. Remarkably, transplantation of cell populations enriched for the putative human organizer into frog embryos induced a secondary axis. Our research demonstrates that the human organizer can be induced in vitro and paves the way for the study of pattern formation and the initial regulation of body axis establishment in humans. STEM Cells 2011;29:600–607
- Top of page
- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
In their milestone experiments, Spemann and Mangold [1] identified the cells at the dorsal lip of the amphibian blastopore as responsible for the patterning of the anterior-posterior and dorsal-ventral axes of the developing gastrula embryo and termed them the "organizer." This role was realized through their ability to induce dorsal structures, and particularly neural precursors, when transplanted heterotopically. On the molecular level, the expression of several genes is characteristic of the amphibian organizer during gastrulation, the most prominent of which is the paired-type homeodomain transcription factor goosecoid (GSC). The induction of the organizer cells is regulated by the costimulation of the transforming growth factor beta (TGFβ) and the WNT pathways [2]. Consequently, the organizer itself secretes numerous proteins that form intricate regulatory networks and eventually induce the TGFβ pathway and inhibit the BMP and WNT pathways. Among these are Nodal related proteins [3], which are members of the TGFβ family, and Dickkopf 1 (DKK1), which inhibits the WNT pathway. Cerberus 1 (CER1), another prominent marker of the organizer, is a tripartite inhibitor of all three pathways.
Numerous studies have shown high evolutionary conservation of the function and molecular basis of the organizer among vertebrates, specifically in fish, frog, chick, and mouse [4]. However, significant differences do exist. For instance, in mouse, the gastrula organizer activity seems to have been divided between two GSC expressing regions in the cup shaped early embryo, as opposed to the single structure in amphibians [5–7]. As the human gastrula differs from its murine counterpart in many aspects, the human organizer is expected to exhibit unique features. However, obvious ethical restrictions prevent the direct study of early human embryos.
Human embryonic stem cells (hESCs) are pluripotent cells derived from the pregastrula embryo [8]. In vitro, hESCs can be aggregated to form embryoid bodies (EBs) [9]. This induces their differentiation into progenitors and derivatives of the three embryonic germ layers, following a sequence similar to normal embryonic development, but without pattern formation [9–12].
Here, we study the differentiation of hESCs into cells that harbor markers and characteristics of the gastrula-organizer. Through genetic labeling of hESCs for GSC expression, we could purify and determine the gene profile of the putative human organizer cells. Remarkably, transplantation of GSC expressing cells into frog embryos induced secondary axes, suggesting human EBs harbor organizer-like cells.
- Top of page
- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
H9 hESCs were cultured using standard procedures [10]. In vitro differentiation into EBs was performed by withdrawal of bFGF from the growth media, and factors were added with the initiation of EBs formation. ActivinA and DKK1 were purchased from PeproTech (Rocky Hill, NJ, www.peprotech.com). SB-431542 was purchased from Tocris Bioscience (Bristol, U.K, www.tocris.com) and from Cayman Chemical (Ann Arbor, MI, www.caymanchem.com). Total RNA was extracted using RNeasy Mini or Micro (Qiagen, Valencia, CA, www.qiagen.com). RNA was reverse transcribed by random hexamer priming (Promega, Madison, WI, www.promega.com) and TaqMan probes (Applied Biosystems, Warrington, U.K., www.appliedbiosystems.com) was used for real-time polymerase chain reaction (PCR). Immunostaining of cryosectioned EBs was performed using primary antibodies against GSC (Abnova, Taipei city, Taiwan, www.abnova.com), FOXA2 (Abcam, Cambridge, MA, www.abcam.com), NOGGIN (Santa Cruz Biotechnology, Santa Cruz, CA, www.scbt.com), NODAL (Santa Cruz Biotechnology), CER1 (Sigma-Aldrich, Saint Louis, MO, www.sigmaaldrich.com), and β-CATENIN (Cell Signaling, Danvers, MA, www.Cellsignal.com). Cy3 and Cy2 conjugated antibodies were used as secondary antibodies, and nuclear staining was performed with Hoechst 33258 (Sigma). For DiI labeling, we used Vybrant multicolor cell labeling kit (Invitrogen, Carlsbad, CA, www.invitrogen.com). Global gene expression analysis was performed using Affymetrix Gene ST1.0 microarray. Data was normalized, and genes upregulated in GSC-GFP+ cells were identified by being of higher expression in the GFP+ population in both 2- and 3-day EBs by more than 1.5 log values and by having expression above array average in the GFP+ cells. Dendrogram was made using the Expander integrative program suite [13]. Genetic labeling for GSC and CER1 expression was made using the recombineering technique [14], and a fragment containing the sequence for eGFP and the neomycin resistance was used to replace the GSC open reading frame (ORF) in RP11-179A9 BAC or the CER1 ORF in RP11-696E8 BAC (BACPAC resources, Oakland, CA, http://bacpac.chori.org). The first modified BAC was linearized by digestion with AgeI (New England Biolabs, Ipswitch, MA, www.neb.com) and the second with Kpn2I (Fermentas, Hanover, MD, www.fermentas.com). After restriction, the BAC was electroporated into H9 cells. Fluorescent activated cell sorting (FACS) analysis and sorting was performed using FACSCalibur and FACSAria Cell-Sorting Systems (Becton Dickinson, Franklin Lakes, NJ, www.bd.com), respectively, after EB dissociation. Einsteck procedure was performed in Steinberg's solution. A small puncture was made ventrally in late blastula Xenopus embryos using delicate tweezers, and the chorion was removed. EBs were washed thoroughly to remove residual Activin A and dissected to expose their interior. A piece was introduced into the embryo's blastocoel through the puncture. In the refined Einsteck procedure, sorted cells were resuspended in phosphate buffered saline (PBS) with 50% Matrigel (BD Biosciences) to an estimated concentration of about 150,000 cells per μl. Using a fine glass needle, each embryo was injected with 20–40 nl of the mixture. Treated embryos were allowed to develop further in 17°C. 5-bromo-4chloro-3-indolyl-phosphate (BCIP) or magenta phosphate was used to identify digoxigenin labeled RNA probes that hybridized to endogenous frog transcripts by in situ hybridization, as described previously [15].
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- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
TGFβ and WNT Pathways Induce Differentiating hESCs to Express Organizer Related Genes
Numerous studies have shown high evolutionary conservation of the function and molecular basis of the organizer among vertebrates, particularly in fish, frog, chick, and mouse [4]. Similar to the Xenopus organizer, it seems that both TGFβ and WNT pathways have a crucial role in mouse organizer formation [16–20]. To test whether these two pathways affect the establishment of the human gastrula organizer, hESCs were harvested and allowed to aggregate into EBs in the presence of Activin A and LiCl, activators of the TGFβ [21] and WNT [22] pathways, respectively. mRNA was extracted 2 days after EB formation, and changes in organizer-related gene expression were analyzed using real-time PCR. Indeed, most of the genes examined were affected by the treatments, and three groups of genes could be discerned. The first group, containing genes that responded to administration of Activin A only, included GSC, CER1, LIM homeobox 1 (LIM1), hematopoietically expressed homeobox (HHEX), and NODAL [23] (Fig. 1A). The second group, which included BRACHYURY [24, 25] and CXCR4 [26], was upregulated by either Activin A or LiCl (Fig. 1B). The third group, which was represented by CHORDIN [27], showed no response to either treatment (Fig. 1C). As the role of WNT pathway in organizer formation is well-established, we speculated that baseline levels of endogenous WNT activity mask the activation of the pathway by exogenous factors. To test this hypothesis, we formed EBs in the presence of recombinant DKK1, a WNT inhibitor. This indeed brought about downregulation of genes from all three groups, including GSC, BRACHYURY, and CHORDIN (Fig. 1F). This shows that in hESCs, too, the WNT pathway is necessary for the induction of organizer related genes. Addition of the TGFβ inhibitor SB-431542 similarly downregulated organizer related genes (Fig. 1F). Altogether, the effect of TGFβ and WNT signaling on hESC differentiation indicates that the mechanisms underlying the induction of the human organizer are similar to those of other vertebrates.
GSC As a Marker for Putative Human Gastrula Organizer Cells
Of the genes examined, the paired-type homeodomain transcription factor GSC is expressed in the amphibian dorsal lip and in both anterior primitive streak (APS) and anterior visceral endoderm (AVE) of the mouse [5, 6, 28] and is therefore considered as a prominent marker of the gastrula organizer [29, 30]. Here, we show that many of the GSC expressing cells also express other proteins related to the organizer, such as Forkhead box A2 (FOXA2), CER1, NODAL, and NOGGIN (Fig. 1E). GSC mRNA was substantially upregulated by Activin A (more than 10-fold when compared with untreated EBs, p = .04), and immunostaining corroborated the effect of Activin A on GSC at the protein level (Fig. 1D). Therefore, we chose to focus on GSC as a marker for putative human gastrula organizer cells and on Activin A as an inducer of this system.
Genetic Labeling and Characterization of GSC Expressing Cells
To better characterize the GSC expressing cells, hESCs were genetically labeled to monitor GSC expression. A bacterial artificial chromosome (BAC) in which the GSC ORF was replaced with enhanced green fluorescent protein (eGFP) was introduced into hESCs to establish GSC-GFP hESC reporter clones (Fig. 2A). EBs from GSC-GFP hESCs treated with Activin A contained several clusters of GFP+ cells (Fig. 2B) which, in contrast to mouse ESCs, did not seem to localize to a single pole within the structure [31]. Cell sorting of dissociated 2–3 days old EBs treated with Activin A showed that GSC mRNA levels were more than 7.5 times higher in GFP+ cells when compared with GFP− cells (p < .005; Fig. 3B), confirming the reliability of the F3 genetic labeling with the BAC construct.
FACS analysis of dissociated EBs 2 days after aggregation shows that GSC is indeed induced by Activin A (Fig. 2C). In the absence of Activin A, approximately 3% of the cells showed GFP expression. On addition of Activin A at increasing concentrations, GFP+ cells became gradually more abundant until a plateau was reached at 60 ng/ml Activin A, with an average of approximately 20% of the cells positive for GFP (Fig. 2D). DKK1 addition abrogated Activin A induced GFP expression, suggesting that endogenous WNT activity facilitates GSC expression (Supporting Information Fig. 1B, 1C).
Activin A treated EBs started expressing GFP in a small number of cells beginning 1 day after EB aggregation, and a peak (>40%) was reached after 4 days. By the 7th day, the number of GFP+ cells declined dramatically (Fig. 2E). In control EBs, the number of GFP+ cells remained low throughout the entire week (<3%). The transient expression of GSC in vitro corresponds to a transient population of cells, which function at the earliest stages of human gastrulation. To obtain putative gastrula organizer cells, we chose to focus on GSC-GFP+ cells that appear early after the induction of differentiation, that is, no more than 3 days after EB formation.
Transcriptome Analysis of GSC+ Cells Derived from Differentiating hESCs Reveals the Molecular Constitution of the Organizer
To elucidate the identity of the GSC expressing cells, we performed whole transcriptome microarray analysis on mRNA extracted from sorted GFP+ and GFP− cells isolated 2 or 3 days after EB formation, in the presence of Activin A. Seventy-five genes were found to have substantially higher expression levels in the GSC-GFP+ cell population, and for several transcription factors and secreted proteins, this was verified by real-time PCR (Fig. 3B). We compared the expression of genes enriched in the GSC-GFP+ cell population with the expression of their frog homologs in the dorsal side of stage 10.5 Xenopus embryos [32]. This analysis showed that both the hESC derived GSC-GFP+ cells and the frog dorsal region express the classical organizer markers (GSC, DKK1, and CER1), alongside other organizer related genes (Supporting Information Fig. 2). Furthermore, analysis of available literature revealed that the genes enriched in the GSC-GFP+ cell population show an overwhelming enrichment of organizer-related genes. Nine of eleven (82%) transcription factors (Fig. 3A, I) are known from other vertebrates to have a role related with organizer function or to be expressed in the organizing regions. These include GSC itself, FOXA2, MIXL1, LIM1, SOX17, EOMES, and others. EOMES is also known to be involved in epithelial to mesenchymal transition, a hallmark of organizer function, and indeed the GSC-GFP+ cell population showed higher N-CADHERIN expression and lower E-CADHERIN levels when compared with the GFP− cells obtained from 2 days old EBs. Among the secreted molecules strongly coexpressed with GSC in the GFP+ cells (Fig. 3A, II), 5 of 12 (41%) are organizer-related and so are 5 of 19 (26%) receptors (Fig. 3A, III). The Xenopus organizer is known to secrete inhibitors of both the WNT and the BMP pathways, in parallel to activation of the Nodal/Activin pathway. Indeed, the genes upregulated in the GSC-GFP+ cells include two members of the DKK family of WNT inhibitors (DKK1 and DKK4), alongside with CER1, a tripartite inhibitor of WNT, BMP, and TGFβ [33]. Also upregulated in the GSC-GFP+ cells are NODAL, the Nodal/Activin activator, and its coreceptor, teratocarcinoma-derived growth factor 1 (TDGF1). Although it activates the competing BMP pathway, we found BMP2 to be upregulated in the GSC-GFP+ cells. This may correlate with its expression, together with anti-dorsalizing morphogenic protein (ADMP), at the Xenopus organizer, where it seems to have a role in limiting organizer expansion [34–36].
The genes upregulated in the GSC-GFP− cells represent a more heterogeneous population, as they cannot be attributed to any particular known cell type (Fig. 3A, IV). Follistatin is the only organizer related molecule enriched in them. However, during early frog development, Follistatin seems to be excluded from the GSC expressing cells within the organizer [37], further emphasizing the striking resemblance between amphibian and human GSC+ cells. Gene ontology based functional annotation [38] reveals a significant enrichment of genes related to neural development within the GSC-GFP− cells, including CRABP1, DLK1, OLIG3, PAX3, and others. As neural induction is a hallmark of organizer function, it is possible that the GSC expressing cells had induced this fate on neighboring hESCs during their initial differentiation.
To further verify the molecular identity of the GSC+ cells, we established additional lines of hESCs that were now labeled for expression of CER1, a secreted molecule related with the organizer (Supporting Information Fig. 3). Although not identical to GSC+ cells, the CER1+ cells show very high similarity with their molecular composition. As GSC expression is probably shared by a few cell populations, this similarity shows that the subpopulations are highly similar, and most probably differ in the expression of only a low number of genes.
Transplantation of GSC-GFP+ Cells into Frog Gastrula Induces a Secondary Axis
After demonstrating that hESCs can be induced to differentiate into cells with the molecular signature of the gastrula organizer, we wanted to see if the same culture conditions could also establish the organizer function. Previously, Blum et al. [29] showed that transplantation of the GSC expressing distal tip of gastrula stage mouse embryos into Xenopus embryos induced partial secondary dorsal axes—thus identifying the tip as the area containing the murine gastrula organizer. To demonstrate a human gastrula organizer function, differentiated hESCs enriched for GSC expressing cells were transplanted into late blastula (st. 8–8.5) Xenopus embryos using the Einsteck procedure [39]. A small puncture was made in the ventral animal cap, and a fragment from an Activin A treated EB was first thoroughly washed to remove residues of the factor, and then inserted through the incision into the blastocoel cavity (Fig. 4A, I). The embryos were allowed to develop for an additional 24–36 hours. In situ hybridization was performed with probes specific for various frog axial markers including NCAM (neural tube). Transplantation of early (1–2 days old) EBs treated with Activin A induced a secondary axis in more than 25% of the transplanted embryos (15 of 57) (Fig. 4A, II–VI; Supporting Information Table 1A). If the transplanted EB was grown in the absence of Activin A, the frequency of axial structure induction was significantly lower (5 of 51, p = .045, Fisher's exact test), similar to the results obtained from sham operated embryos (3 of 46, 6.5%). The latter is probably caused by scarring inflicted by the intrusive Einsteck procedure. This, we assume, may also be the cause for some of the locally restricted patches of staining observed in several cases after in situ hybridization.
To better investigate the organizing activity of the GSC expressing cells within the EBs, we chose to refine the Einsteck procedure (Fig. 4B, I). For that purpose, GSC-GFP genetically labeled cells were aggregated into EBs in the presence of Activin A and allowed to differentiate for 2 days. The EBs were dissociated and either GFP+ or GFP− sorted cells were injected through a fine needle into the blastocoel cavity of Xenopus embryos (approximately 3,000 cells per embryo). On reaching st. 25–26, the embryos were examined for the induction of secondary axial structures by in situ hybridization with a probe specific to the frog NCAM (Fig. 4B, II–VII). Embryos injected with GFP+ cells contained an induced secondary axis in 22% of the cases (11 of 48 and Supporting Information Table 1B), whereas the GFP− injected embryos showed axial inductions in less than 5% of the embryos (2 of 45), a difference which proved statistically significant (p < .015, Fisher's exact test, two independent experiments). Notably, many of the structures induced by the GFP+ cells were composed of clear elongated axes, none of which was present in the embryos injected with the GFP− cells. Histological examination showed that the GFP+-cells could induce an axis in which the epithelial cells fold to form the typical tubular morphology of a neural tube (Fig. 4B, VI,VII).
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- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
Gastrulation involves both the differentiation into the three embryonic germ layers and the patterning of the main body axes. Previous studies in hESCs have related mostly to the first aspect and regarded GSC as one of several markers for definitive endoderm [40, 41]. Here, for the first time, we isolate human GSC expressing cells formed during the earliest stages of hESCs differentiation and examine their role as the putative human organizer. We show that these cells are induced by molecular pathways known to induce the organizer in other vertebrates, and that they express genes related to the establishment of the early embryonic axes. Finally, we show that these cells posses the function of the gastrula organizer, as they can induce the formation of a neural tube when transplanted ventrally to blastula stage frog embryos.
The effect of TGFβ and WNT signaling on hESC differentiation indicates that the mechanisms underlying the induction of the human organizer are similar to those of other vertebrates. The amphibian organizer can be subdivided into more specialized organizers which pattern the head, the trunk, and the tail [42]. In mouse embryos, the gastrula organizer activity seems to have been divided between two GSC expressing regions in the cup shaped early embryo (approximately 6.5 days post coitum). In the posterior part of the embryo, the organizing center is the APS [5] that contains mesendoderm and is the physiological equivalent of the amphibian dorsal lip of the blastopore. In the embryo's anterior region, the extra embryonic cells at the AVE behave as an organizing center [6, 7]. Accordingly, our results point to the fact that the GSC+ cell population is heterogeneous (Fig. 1). However, as observed by the similarity between GSC+ and CER1+ cells, these subpopulations are expected to differ only slightly. Future work should focus on identifying the components of the human organizer, and intraspecies transplantations can provide means to examine their distinct roles.
A comprehensive transcriptome analysis of isolated GSC+ cells revealed that many organizer related genes are coexpressed with GSC. The varying proportion of known organizer related genes among different functional groups observed in genes upregulated in GSC+ cells conforms with the current knowledge regarding axis formation. Most of our information is of transcription factors involved in the process, as these stand at the top of the process, and easily show a phenotype when manipulated. Similarly, as the organizer secretes strong inducers to perform its function [43], many of its secreted molecules are known. However, little is known about the receptors on the surface of organizer cells. These receptors are assumed to be involved, among other things, in the regulation of organizer localization and in directing cell migration during axis formation. Indeed, the receptors upregulated in GSC+ cells include, among others, PLXNA2 [44] and SEMA5A [45], known to take part in cellular position determination during axonal guidance. The genes upregulated in the GSC expressing cells may, therefore, posses a yet unknown role in the patterning of the mammalian and particularly the human axis.
When injected ventrally into frog blastula-stage embryos, GSC+ cells induced differentiation of a secondary neural tube. Although the axes induced by direct injection of GSC+ cells seem more complete than those obtained by EB transplantation, they are still partial in comparison to those reported when frogs are transplanted with organizers of their own species. However, it should be noted that in interspecies transplantations, most secondary axes are incomplete as in the case of the mouse distal tip [29]. We assume that the suboptimal conditions for hESCs growth in the frog embryo hampered their further proliferation and differentiation and thus may have also reduced their axis inducing ability. However, the induction of host cells to posses molecular and morphological hallmarks of a neural tube clearly demonstrates that the human GSC+ cells act as a gastrula organizer. Functional manipulation of either the host embryo or the transplanted human organizer cells (using Morpholinos or small molecules) should allow further understanding of the molecular interactions, which facilitate the organizer.
Understanding the mechanisms regulating early shape induction in the embryo is important for both the ability to promote organ differentiation in vitro and for deciphering early human embryogenesis. For the first time, we demonstrate that hESCs can differentiate into cells with the molecular signature and function of the gastrula organizer and present an experimental model-system that should allow the study of early body plan patterning in the human embryo. Molecular analysis of GSC+ cells and their ability to induce a secondary axis in frog embryos emphasize the extraordinary evolutionary conservation in organizer function between human and other vertebrates. Future work should focus on characterizing the subpopulations within the GSC+ cells and on the roles of the genes, we identified as organizer-related, in axis formation.
- Top of page
- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
Human embryoid bodies derived from embryonic stem cells express markers characteristic of the gastrula-organizer. Genetic labeling of the organizer cells enabled their purification and molecular characterization. Interestingly, transplantation of these cells into frog embryos induced a secondary axis. The discovery of the human gastrula organizer within human embryoid bodies paves the way for the study of pattern formation and body axis establishment in humans.
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- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
We thank Drs. Naomi Melamed-Book and Adi Mizrahi for assisting with microscopic analysis; Dr. Michael Zeira and Dan Lehmann for assisting with cell sorting; Dr. Graciella Pillemer for assisting with in situ hybridizations; and Drs. Danny Kitsberg and Yoav Mayshar for critically reading the manuscript. N.B. is the Herbert Cohn Chair in Cancer Research. This research was supported partially by the ISF-Morasha Foundation (grant no. 943/09; to N.B.), the Israel Science Foundation, and the Wolfson Family Chair in Genetics (to A.F) and a grant from the Legacy Heritage Fund of New York (to N.B).
Disclosure of POTENTIAL CONFLICTS OF INTEREST
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- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
The authors indicate no potential conflicts of interest.
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- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
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Supporting Information
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- Abstract
- Acknowledgements
- Disclosure of POTENTIAL CONFLICTS OF INTEREST
- Supporting Information
Additional supporting information available online.
|Supplementary Figure 1: Endogenous WNT activity facilitates baseline activation of GSC in early EBs. a. Effect of Activin A and DKK1 on the percentage of GFP+ cells obtained from dissociated 2-3 day-old GSC-GFP EBs. Bars show fold induction compared to the control, and error bars represent standard errors. n=3. b-d. FACS analysis of dissociated 3 day old EBs. b. Control EBs. c. EBs treated with 67ng/ml ActivinA. d. EBs treated with 67ng/ml ActivinA and 200ng/ml DKK1.
|Supplementary Figure 2: Major organizer related genes upregulated in both GSC-GFP+ cells and dorsal pieces obtained from st.10.5 Xenopus laevis embryos. the graph presents the log values of the change in expression of the genes. For each gene, its enrichment is presented in GSC-GFP+ vs. GSC-GFP- hESC derived cells; and in dorsal vs. ventral pieces obtained from st. 10.5 frog embryos.
|Supplementary Figure 3: Genetic labeling and molecular analysis of CER1+ cells. a. A scheme of a BAC containing the CER-GFP reporter construct. The two exons of CER1 ORF were replaced with the sequence coding for eGFP adjacent to the neomycin resistance gene under constitutive SV40 regulation. b. Early EBs made of CER-GFP cells show clusters of GFP expressing cells. Left - bright field; Center - dark field; Right - overlay of bright and dark fields. c. FACS analysis of dissociated 2 days old EBs. Left- CER-GFP cells; Right - CER-GFP cells treated with 67ng/ml Activin A. d. Effect of Activin A concentration on the percentage of GFP+ cells obtained from dissociated 2 days old CER-GFP EBs. Error bars represent standard errors. Asterisk indicates p-value≤0.05 under 2 sided paired t-test (n=2). e-g. CER-GFP cells were aggregated into EBs in the presence of 67ng/μl Activin A and sorted after 2 days. e. Relative CER1 expression in the sorted populations, as deduced from real time PCR. f. Heat map analysis of gene expression in GFP+ and GFP- populations obtained from Activin A treated EBs of GSC-GFP and CER-GFP cells, 2 or 3 days after aggregation. The main organizer related genes upregulated in the GSC+ cell population show a similar expression pattern in the CER1+ cells when mRNA extracted from them is hybridized to Affymetrix Gene ST1.0 microarray. g. Gene expression pattern of CER1+ cells is highly similar to that of GSC+ cells. A dendogram analysis by Pearson correlation was performed on the genes which showed highest substantial differences (>1 log difference, either upregulated or downregulated) between GSC+ and GSC- cells obtained from 2 day old EBs treated with Activin A. CER1+ and GSC+ cells from 2 day old EBs showed the highest similarity. Adult tissue average expression levels are based on data published by Affymetrix.
|Supplementary Table 1: a. Analysis of the number of secondary axes obtained from transplantations of EBs into frog embryos. b. Analysis of the number of secondary axes obtained from injection of GSC-GFP cells into frog embryos
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The area of interest is a small, fenced-in backyard located in a residential building complex. The area has not been maintained by its previous owner. It is covered in weeds with no grass, and two full grown trees are present. No mulch bed is present. My goal is as follows:
1.)Strip the weedy/grass mix
2.)Bring in fresh lawn soil
3.)Make a bedding area around the perimeter of the fence
4.) Plant shrubs/plants that are most appropriate for the environmental and soil conditions
5.) Put down a preventative tarp in the bed to keep weeds from popping up
6.) Lay mulch
7.) Add a fertilizer to the soil to ensure sod growth
5.) Install sod (estimated 25 2x4 strips of sod
6.) Finally, power wash the small deck and clean up all the stripped grass and dirt.
Another thing to keep in mind to help make a more accurate estimate is that the only entrance to the backyard is through the building, as the yard is surrounded by neighbors and there is not an opening in the fence. That would mean that materials and clean up would have to be done manually into and out of the building.
Fence and landscaping on roof.


text:
Three to See: Suggestions for every day of the Big Gig
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Drunk fest commences.
This Sunday -- 7/7, A.M. Taxi is is MUST see.
Kathy Griffin, Widespread Panic among new concert announcements
Milwaukee movie guide: What's playing where
Now streaming: What's new, what's leaving streaming services this week
Concert Announcement: Jenny Lewis' new supergroup NAF opening for M. Ward at Pabst Theater
Local Beat: Seven top summer concerts featuring Milwaukee bands
Restaurant-bar Wall Street Drink Exchange due in August in Elm Grove
WITI-TV leads overall, barely, in May sweeps; WISN-TV newscast tops at 5, 6 p.m.
Milwaukee movies off the grid: May 27-June 2
Beer Here: Kicking off summer by the pint
20 top Milwaukee concerts this summer
TAP'd In Podcast: Talking top summer concerts with Lil Wayne, Kenny Chesney, more
Top shows this week: WebsterX, Curren$y, Milk Carton Kids and more
Movies: Opening this week — 'X-Men: Apocalypse,' 'Alice' sequel, more
On Tap Today: Bavarian Bierhaus grand opening
Documentary 'Milwaukee 53206' to have world premiere here
Throwback Thursday — Comics Edition: 'Bloom County' gave us Opus, Bill the Cat — then took them away
Concert Announcement: Julieta Venegas' Riverside Theater show Saturday canceled
The 24-tap ABV Social opens June 1 at the Mayfair Collection in Tosa
Concert Announcement: Lakefront Festival of Arts lineup announced
Skylight Music Theatre artistic director Viswa Subbaraman to step down
Concert Review: The 1975 new and improved at the Rave's Eagles Ballroom
Latest Festival News
- Free concerts in the Milwaukee area this summer
- Church festivals, fairs and other events in the Milwaukee area this summer
- County fairs in the Milwaukee area this summer (2)
- Guide to Milwaukee area summer festivals 2016
- Pitbull headlining Summerfest June 30 (8)
- State Fair announces lineup for Bank Mutual Amphitheater


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They recognized Kosova which means they should also recognize Nagorno, Abkhazia and South Ossetia.
They recognized Kosova which means they should also recognize Nagorno, Abkhazia and South Ossetia.
|Thumbs Up/Down
|Received: 21/2
Given: 0/0
Actually, why doesn't even Armenia recognize it?On December 10, 1991, as the Soviet Union was dissolving, a referendum held in the NKAO and the neighboring Shahumian region resulted in a declaration of independence from Azerbaijan as the Nagorno-Karabakh Republic. The country remains unrecognized by any UN-member state, including Armenia. Only Transnistria, South Ossetia, and Abkhazia, UN non-members, recognize the state.
|Thumbs Up/Down
|Received: 51/10
Given: 1/0
|Thumbs Up/Down
|Received: 5/0
Given: 4/0
State recognition is a political game. The US (or any other state for that matter) only recognises a state if it is in its interest to do so.
|Thumbs Up/Down
|Received: 39/9
Given: 0/0
I personally think most of that territory has little to do with Azerbaidjan and some sort of independence solution is most favourable.
|Thumbs Up/Down
|Received: 2,279/75
Given: 2,099/113
This Garuda column of Vasudeva (Vishnu), the god of gods, was erected here by Heliodorus, a worshiper of Vishnu, the son of Dion, and an inhabitant of Taxila, who came as Greek ambassador from the Great King Antialkidas to King Kasiputra Bhagabhadra, the Savior, then reigning prosperously in the fourteenth year of his kingship. Three important precepts when practiced lead to heaven: self-restraint, charity, conscientiousness.
-Heliodorus Pillar 113 BCE
|Thumbs Up/Down
|Received: 405/194
Given: 353/256
Because that region has nothing to offer US.
|Thumbs Up/Down
|Received: 2/1
Given: 1/0
Because South Ossezia, Nagorno-Karabakh and Abkhazia haven't the petroil
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|veradriane .
May 19
An Upfront Honest Review of The Great Gatsby
So a week ago, I got a glimpse at "The Great Gatsby". Didn't watch the full movie back then, tho, as getting up early for a meeting was my main goal that day. And before you start judging me on my obvious lack of film taste, ... 0
|rajatsingh .
Mar 21
Benefits of Using the Braided Copper Wire
You will find wires of different shapes and sizes and it is important to weigh the pros and cons of each variety when deciding on the type of wire to use in making an electrical product. So, the braided copper wire is o... 0
|Ckervin .
Jan 29
[url="http://www.flixist.com/blogs/Ckervin/the-future-is-looking-pretty-rough--220323.phtml"]The future is looking pretty rough[/url] in these films!
|Ckervin .
Jan 29
Well, this one will be interesting!
|rajatsingh .
Jan 12
Despite Certain Drawbacks - Quality Aluminum Wires Continue To Be In Demand
Both aluminum and copper are used as conductor material. But for users the choice is governed by a compromise between electrical properties, mechanical properties and price. Historically, copper had an edge over... 0
|scoatparkey .
Jan 05
What Makes VPS Hosting in India Work?
Well, shared hosting is indeed the cheapest hosting solution available today. However, there would be a time when you would outgrow shared hosting and will not be able to put up with its resource limitations. This is when VPS... 0
|scoatparkey .
Jan 05
What Makes VPS Hosting in India Work?
|Retrofraction .
Dec 19
Well super excited to see Hateful Eight sometime soon, but does anyone have and recommendations for some good indie films?
|rajatsingh .
Dec 10
How is buying Nickel Copper Wire Beneficial?
More and more electrical good manufacturing companies are using nickel copper wire because of the intrinsic advantages associated with it. Greatest benefit is surely its capability of resisting high corrosion. Moreover, elect... 0
|imran haider .
Dec 02
Hire the Services of Quality Server Colocation Services Providers in India
Owning a data center,even a small one,is beyond the technical and financial capabilities of large number of organizations. Forms may have to spend a large amount of money ain building their own data centre and hiring expert I... 0
|kajolsadef .
Nov 08
vdzczsddc 0
|Charlie Cooke .
Nov 05
SPECTRE Review
James Bland The first three Daniel Craig James Bond films exist as a kind of trilogy, telling the story of Bond's evolution from blunt killing machine to suave, super-spy. By the end of Skyfall, Bond was in a recognisable p... 0
|Ashley Elizabeth .
Sep 03
Four types of office desks That Can Increase Productivity
The right office furniture can increase employee productivity and improve quality of work. The interior of an office should be designed in a way to provide maximum comfort and warm environment for employees to work. The most ... 0
|Ashley Elizabeth .
Sep 02
Solar Panels in UK
The sun is a powerful source of energy. You'd be surprise to know that humans are able to harness just 0.001 percent of sun's energy. However, in recent years, the use of solar energy for electricity generation ha... 0
|Ashley Elizabeth .
Aug 26
Types of Surety Bonds To Choose From
Surety bonds are the best way to prevent/manage risks in construction projects. Surety bonds ensure that projects will be completed as per specified terms & conditions and on time & budget by the contractor. A contra... 0
|Niko Niko .
Aug 23
|Niero .
Aug 11
Testing Quickposts! You can now leave a short note on Flixist about anything, and continue/convert it to a blog later
|Retrofraction .
May 04
Did you like Captain America 1 or 2 better?
I know this is probabbly the wroung question to be asking here. Both movies are AAA buddget Hollywood at its best. But if you had to sit down and watch one of them over again which one would you rather watch again? Perso... 0
|TheeHeadAche .
Apr 13
Don Hertzfeldt�s World of Tomorrow is sublime.
The splendid tragedy of Don Hertzfeldt's projected future in World of Tomorrow cuts to the core of many sentiments and fears within an individual today. In fifteen minutes he manages to craft a story so thoughtful, fun... 0
|TheeHeadAche .
Mar 16
My Adventure into Film Perversion.
I am a fan of film. I am also a fan of perverse film. I'm such a fan of these things, that I think my compass for what is cinematically perverse or strange may be off, but thats not the problem. The reason I bring ... 0
|TheeHeadAche .
Mar 13
Why I LOVE East of Eden
One good brother. One bad. Elia Kazan's 1955 film, East of Eden, is a masterpiece. Top-notch performances, smooth pacing and flawless shot composition make this my favorite film of all time. Kazan's status a... 0
|Xzyliac .
May 19
The Flixist XCast 3: X3/Origins: Wolverine
Title:�X-Men: The Last Stand Release Date:�May 26, 2006 (Theatrical) Director:�Brett Ratner Writers:�Simon Kinberg and Zak Penn Starring:�Hugh Jackman, Patrick Stewart, Ian McKellan, Kelsey Grammer, Anna Paquin, Rebecca Rom... 0
|Xzyliac .
Apr 01
The Flixist XCast Episode II: X2: X-Men United (2003)
Title:�X2: X-Men United Release Date:�May 2, 2003 (Theatrical) Director:�Bryan Singer Writers:�Michael Dougherty, Dan Harris, David Hayter Starring:�Hugh Jackman, Patrick Stewart, Ian McKellan, Anna Paquin, Rebecca Romijn-S... 0
|Xzyliac .
Mar 17
The Flixist XCast 2 Records Tonight
I've tried to write this blog twice in the last 24 hours. Third time's gotta be the charm. Now that the first episode is out of the way (and the reaction has been great), we're moving on to XCast #2! Centered around 2003's... 0
|Xzyliac .
Mar 03
The Flixist Xmencast Episode I Records Tonight!
After working out the scheduling kinks, we're finally recording the first Xmencast tonight! Are you pumped? I'm pumped. Let's get pumpeder. Tonight we'll be discussing the one that started it all. The first X-Men film star... 0
|Xzyliac .
Feb 27
The Flixist Xmencast: The Genetically Superior Podcast
Dang, it's been a while. Hello Flixist Nation. Xzyliac is the name and even if you forgot about me, trust me I did not forget about you. And like the Phoenix Force itself, I have returned bearing what will most likely be a ... 0
|TheManatee .
Jan 04
Gratuitous Violence and Awkward Silence!
I had no idea what to expect when heading in to the theatre to watch the Ryan Gosling movie Drive. I had heard it was a terrible movie from some less than reputable sources (namely a couple friends with wildly different taste... 0
|TheManatee .
Dec 06
The (hopefully) everlasting zombie genre
I feel the zombie genre, over the past year or so, has sort of burnt itself out and is starting to slow down, which leaves me disappointed. Despite what people say I will always have a soft spot for the undead; from movies li... 0
|Antwhan .
Nov 15
8th Annual Teaneck International Film Festival This Weekend, My Film's In It!
Hey Everybody! My Thesis Film is playing this weekend Sunday evening during the http://www.teaneckfilmfestival.org/� It's Part of the NJ/NY Short Film Series at 6:15 pm on this coming Sunday, November 17th! If any of you are in the area, I'd love it if you came out to catch the film, alongside some other pieces that should be good fun as well! 0
|TheManatee .
Nov 13
Is Marvel's Agents of S.H.E.I.L.D. slowly getting better?
So I started watching this show trying to keep my expectation low, but the first episode got me a little excited as it built off of the extremis idea from Iron Man 3. Since then I feel it has tried to check off as many clich... 0


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ProClub Line Multi Power Rack
The SMR1000 commercial multi rack system is the perfect machine for providing all size users a safe and comfortable platform to maximize squats and pressing motions. ¼" solid steel gun rack style liftoffs provide multiple positions for a comfortable starting position. Rock solid adjustable safeties allow users to workout safely and maximize their load. The 2"x3" 11-gauge steel frame holds up in any heavy use environment. The back frame is extended to allow spotters easy access. Integrated weight storage keeps the plates right where you need them. The SMR1000 has integrated pegs for strength bands for extreme plyometric training. The integrated chinning bar allows users to incorporate pull ups, chin ups, inversion, ab slings and other body weight resistance systems for amazing versatility. The SMR1000 is an uncompromised monster free weight system. The SMR1000 multi-rack system is perfect for heavy use, heavy load environments, like schools, military, police and fire departments.
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A tip from a neighbor led members of the Clark-Skamania Drug Task Force to a Felida house on Sunday morning, where they arrested a 25-year-old man and seized 292 marijuana plants.
It was the thought to be the sixth grow house bust since Operation Gang Green raided 56 houses on Oct. 13, said Sgt. Pat Moore, a Vancouver police officer who is a supervisor with the drug task force.
"The house was being rented," Moore said. "The gentleman had been living here about five months."
Moore said Michael Y. Chen allowed officers into the house at 12103 N.W. 41st. Ave in Messner Estates, a development of 30 houses in the upscale neighborhood. Chen was arrested for manufacturing marijuana, a felony.
One room contained the 292 plants and four others were outfitted with growing equipment, Moore said. "This house was just being set up," he said.
The plants were 6 to 12 inches high, perhaps six weeks from being mature, Moore said. At maturity, each plant can bring $800 to $1,000 on the street, he added.
Moore said there was several thousand dollars in equipment, including light shields, carbon filters with fans, ballasts and timers.
The sergeant said the marijuana and equipment were taken in the task force "raid van." He said grow house busts in Clark County continue and, "It's getting crazy."
Terry Hiller, who lives next door and developed the neighborhood, said his wife, Maureen, suspected a grow operation and alerted police.
"The guy rented it," Terry Hiller said. "Next thing I knew, they had the garage windows blocked out."
He said he only saw Chen about once a month. "He was a really nice kid," Hiller said.
On Sunday morning, Hiller saw Chen at the house, and called Sgt. Moore. Hiller said he went to do errands and his wife called him a bit later and said, " 'There were cops all over the neighborhood.' "
Clark County property records indicate the house is owned by Falcons Run LLC, which has a Washougal address.
Talking about the operation, Moore said wires revealed that power was being stolen from Clark Public Utilities before the electricity reached the house power box.
The plants in the upstairs room were each in pots about six inches deep and six inches wide and thick.
"We're trying to take a zero-tolerance policy on these grows," Moore said. He said many homeowners end up being victims because of damage to the rented houses.
He said the drug task force is not after only marijuana grows. He said members are fighting against all illegal narcotics, including methamphetamine and heroin.
Chen was in the Clark County Jail on Sunday night with no bail allowed.
On Oct. 13, 49 people were arrested when an estimated 300 law officers swept across Clark County and netted about 6,800 marijuana plants. It was thought to be the largest drug bust in Clark County history.


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|product name: Clinelle BLEMISH CLEAR
Blackhead Control Gel (15ml)
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|ative from: 2013-03-16 00:00:00
|product size: 15 ml
|country of origin: USA
|function: Blackhead RemovingDeep CleansingPeeling
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Clinelle Blackhead Control Gel is a control gel formulated to clear blackheads and tighten your pores for a smooth and flawless complexion
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Oleanolic acid, Alpha Bisabolol, Geranium, Essential Oil of Tangerine


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1709 search results for animation
Comcast is buying DreamWorks Animation for $3.8 billion
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Type: Article | Date: Wednesday, May 18, 2016The two-time Oscar winner is in the movie based on the popular game app


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Ceramics Okehampton
3 results for Ceramics in Okehampton
Avon Miniatures20 Brandize PkEX20 1EQ Okehampton 01837 53237
The fine 1 12 scale collectable dollshouse china is handmade by Avon Miniatures in Devon England All of our ceramic ware is made from fine white porcelain which then under goes several firing stages ... pottery, vases, DUCK, bowls, Chelsea plate, CERAMICS, dollhouse, tumblers, Mail Order Catalogue...
The Artful RogersUnit 1, 3 West StreetEX20 1HQ Okehampton 01837319059
... crafts, mostly one off's to suit many different tastes and styles. From intricately painted ceramics to crocheted chairs, from vintage cake stands to crazy lampshades, it's all here, an unusual mix...
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Are you looking for Ceramic Products Manufacturers & Suppliers ?Use our "Request for Quotation" service to get customized offers from companies in Okehampton, tailored for your exact needs!
Versatile CeramicsThe Carlton Suite St James StreetEX20 1DW Okehampton 01837 536 00
... Manufacturing
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Hatherleigh Pottery20 Market StEX20 3JP Hatherleigh 01837 810624
A working pottery with showrooms near the centre of Hatherleigh in the heart of beautiful Devon. Jane Payne runs the Hatherleigh pottery producing practical domestic stoneware. We produce oil burners ... pottery, oil burners devon, michael taylor ceramics, jane payne hatherleigh pottery, oil burners...
Jennie HaleLongham PotteryEX20 4AA Coryton 01822 860287
... make drawings and paintings of the flora and fauna that intrigued me, these would feed my ceramics. When Andrew Osborne and I, where offered the chance of setting up a pottery in a isolated...
Penny SimpsonThe Studio, 44A Court StreetTQ13 8LG Moretonhampstead 01647 440708Closed
Penny Simpson started making pots whilst living in Japan in the 1970s. She was captivated by Japanese food and its presentation colours and textures of pots chosen to complement the food and the...
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Helen Bailes Ceramics Torrington14 The Gallery, The Pannier MarketEX38 8HD Torrington
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Cornish Stoneware CraftGarfield, Tavistock RoadPL15 9HD Launceston 01566 775099
... MNFRS, Pottery Manufacturers, SUPPLIERS, ceramics supplier, pottery products, Pottery Mnfrs...
Monkleigh PotteryBearhaineEX39 5LE Monkleigh 01805 623194
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Holiday Flourishes & Swirls Patterned Paper 8"X8" (36 sheets)
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Using elements from the popular stamp sets Peace Be Still and Fancy Flourishes, Heather Nichols has designed a breathtaking patterned paper collection. Holiday Flourishes & Swirls features sheet music, snowflakes, flourishes, stripes and dots all with a lovely vintage theme. This collection complements Scarlet Jewel, Ripe Avocado, Spring Moss and Fine Linen. 36 sheets per package (6 sheets each of 6 patterns)
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- 2-hour Brooklyn Navy Yard tour
- Learn all about New York's naval and martime history from your guide
- Visit a 19th-century dry dock that's still used today for ship repair
- Check out the country's first multi-story 'green' industrial building and first wind- and solar-powered street lamps
- See historical photographs and listen to audio clips of WW2-era workers, manufacturers and farmers
- Great for history buffs, urban planners, and people interested in sustainable architecture and industry!
Recent Photos of This Tour
What You Can Expect
Customer Reviews
March 2016
The tour guide was OUTSTANDING!
November 2015
GREAT tour, the tour guide was very knowledgeable and warm. Enjoyed all the detailed information he gave us. I was happy to add additional information since I had worked at the Brooklyn Navy Yard in the 50s.
July 2015
This tour was highly interesting and well guided! I wish we could have entered some of the buildings and seen more of the adaptive re-use and sustainability aspects but overall, quite satisfied!
July 2015
Excellent tour
Excellent guide
Will recommend the tour to friends
Really fascinating
Celeste B USA
May 2015
August 2014
Guide was fun,knew the material and it was an enjoyable experience.
January 2014
Fascinating and a worth while trip whole heatedly recommend it to anyone who's new to NYC or just never taken one of these trips ever even if you aren't new here.
December 2013
Very well organized. Excellent tour guide, knowledgeable and pleasant. Presentation well-paced and informative.
April 2013
Excellent tour guide, interesting tour, wish I could have had access to some of the buildings.


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2 Parks near Vista, California
Carlsbad, California 6.96 miles SW
Escondido, California 9.13 miles SE
Oceanside, California 9.51 miles SW
Camp Pendleton Marine Base, California 10.15 miles NW
Valley Center, California 10.72 miles NE
Vista, California (map) | (760) 295-9243
Sites: 60 | Latest Rate: $30.00 | 14 Reviews
|Clean Restrooms:
|Clean Showers:
Oct 2015 - $30.00
This is a standard RV Resort. Sites are concrete and each has a tree and ground cover as well as a patio area and picnic table. (some leveling required as all of the sites are graded down hill, however, the park provides levelers).
Pool is clean and good size. There is also a very large Jacuzzi.
Bathrooms are large and clean. Showers were great!
There is a laundry room with soap machines, etc.
One of the reasons we stayed at this park is that it is located near all major highways and freeway systems. Close to shopping and restaurant's! - jmjack
May 2015 - $20.00
Very friendly young managers. Great wifi, clean restrooms, warm pool and our site was fairly flat concrete. Just one night as passing through but would stay again. Only complaint was office closing at 4pm which is very early. - kiwitravelers
Apr 2015 - $36.00
This is a very nice RV park, but I wouldn't call it a resort. The park is quite clean and well maintained. There are few trees which are too small to provide shade. The roads are paved and the RV pad is concrete. Spaces are fairly tight and there is really not much space for chair or table. The RV pad is level side to side but lengthwise it has such a slope that the park provides leveling blocks. My front tires were a foot off the ground. The pad is barely wide enough for the RV; the slides overhang the planter on one side and the tiny sandy patio area on the other. The campsite has 20/30/50 electric but no picnic table or fire pit. The water is city supplied and fairly decent at about 60 psi. Basic cable is free. Internet wifi is free with a strong signal throughout the park; good enough to stream. The laundry is small with 3 machines, but in good condition. The showers are clean and in good condition; did not use. There is a small pool with spa. The office staff is a helpful young lady who is quite friendly. The park has a few activities, but it's such a small park that events really aren't very feasible. The park is located in a city and close to anything you could want. Noise, even from the commuter train, is minimal. Traffic is always an issue when going anywhere. This is a pleasant park to stay at for doing things in the area, but not as a destination park, unless you're just looking for a place to live. - Wiyndog
Vista, California (map) | (760) 724-1968
Sites: 19 | Latest Rate: $24.00 | 1 Reviews
|Clean Restrooms:
|Clean Showers:
Mar 2016 - $24.00
Contrary to the prior review, this is an Elk member ONLY campground. We had FHU's (30amp) and excellent Wi-Fi is included. - FLtravelers
Jan 2012 - $23.00
An Elks lodge RV park that allows non-members access and takes reservations. Parking is on dirt/gravel on a level lot. Large covered picnic area for groups. Laundry available at lodge building. Narrow sites for rigs with slides on both sides you are only 4 feet from your neighbor when his slides are extended. Big rigs should watch for dragging turning into lodge coming from the north. Host on site. - Bucky Beaver


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Came as a recommendation from the builder. He purchased it from a distributor here in Penna, near Philadelphia.
I bought the reddish color, and it is fading quickly, as well as the high mold marks.
- Forum >
- Lawn & Garden >
- Correct Deck composite
I bought the darkest and it is now light red with gray and mold. Does your builder feel any responsibility for recommending this product?
He didn't feel any responsibility for all the drywall tape that peeled off every corner of the room and ceiling, starting within two weeks after completion of the project, although after 9 months of letters and e-mails, and finally the threat of a law suit, he sent in someone to repair the damage. I didn't even talk to him with regards to this issue.
The person who will likely be our GC for the building of our deck (a SUBSTANTIAL project) just left our house and had recommended Correct Deck CX. Having not heard of this particular brand of composite decking, I did a quick google and found myself in this forum. I'm obviously quite concerned about going this way now, not only for the mold issues, but also for the lack of customer service. My question, though, is are all of you who are experiencing the mold using Correct Deck or Correct Deck CX? Besides the customer service issues (which weigh significantly of course), are you aware of similar mold issues (or any negative issues for that matter) with the CX.
Can't say you haven't been warned. They don't provide customer service on their correct deck material, why would you believe they would provide it on the new and improved(their words) CX?
Correct Deck is perhaps the most irresponsible company I have ever had the misfortune of trying to deal with. They have no concern for their customer won't even reply to phone and e-mail messages. Have you read their 25 year warranty, it doesn't cover anything, not mold, fading, or anything else. You would be so much better off just using wood. This Correct Deck needs more maintenance than my old wood deck and you can't treat it or restain it. It was a total waste of money. Please don't throw your money away. I wish someone would have warned me.
Mojobradley/Jemcge, Thanks for coming back to me so quickly. If you could do it again, which composite product would you go for? I checked out the Ultradeck that someone linked to on this forum (http://www.midwestmanufacturing.com/), but they don't appear to operate in my area (North Jersey). Is there a composite out there that resists stains, mold, mildew, etc? It seems that this is a problem with composites in general.
we are in new jersey the rep i spoke to also said enviroment, balony i have no mold on nothing else in my yard so why just the deck? even if i didnt complain about all the mold the deck was a clay color now its almost white so is that enviroment too like i said there fulll of balony.
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got it today
checked out Moultrie's web site the other day and what a sale. got a D55-IR for $29. they list for $129. any body need a new TC better get there quick and get one before there gone. several other models also
May be late but gonna check em out I love my D-55IR great camera
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