text:
Stephanopoulos Spurns CBS
CBS News courted but couldn't land George Stephanopoulos, who recently signed a new deal, with a nice raise, to stay at ABC News for four more years.
Mr. Stephanopoulos, ABC News' chief Washington correspondent and moderator of ABC News' "This Week," is a rare commodity. He's young, he's articulate and he's telegenic. He knows Washington and he's got connections. He was a key member of Bill Clinton's 1992 presidential campaign team and first administration—but he singed some of his bridges with "All Too Human," the 1999 bestseller he wrote after leaving the White House. Perhaps equally important: He is represented by Alan Berger, the CAA agent who ushered Katie Couric to CBS as a $15-million-a-year "Evening News" anchor/managing editor and occasional contributor to "60 Minutes."
The interest in Mr. Stephanopoulos offers a glimpse into the mind of CBS News and Sports President Sean McManus, who has been identifying young talent he thinks has star potential since he took over CBS News last December with the declaration that "being No. 3 is not acceptable."
Sources close to Mr. Stephanopoulos say Mr. McManus made an aggressive pitch that included talk about multiple plum roles, including Washington bureau chief and chief political analyst, and a high-profile position as the chief on-air political foil for Ms. Couric. He is said to have been told that he would be able to contribute to "60 Minutes."
CBS News sources say Mr. Berger and Mr. Stephanopoulos initiated the talks and that the conversation never turned into an actual offer to Mr. Stephanopoulos. Both sides concede that the topic of bureau chief position arose, but the CBS sources suggested that Mr. McManus was cool to the idea. Janet Leissner has been CBS News VP and Washington bureau chief since 1998.
Both camps agree that Mr. McManus made clear he could not offer a Sunday newsmaker show in the short term because Bob Schieffer is assured that he will be moderator of "Face the Nation" for as long he wishes.
Mr. Schieffer proved himself a most valuable player when he stepped in as interim "Evening News" anchor in March 2005 and proceeded to improve viewership. Mr. Schieffer also was the only party willing to comment on the record about the CBS News-Stephanopoulos talks. "I was amused," he said, "but I understand this is all part of the game."
To wait an indeterminate time for "Face the Nation" would have left Mr. Stephanopoulos without the platform most crucial to building or maintaining TV clout in political and governmental circles.
Since Mr. Stephanopoulos took over "This Week" from Cokie Roberts and Sam Donaldson in 2002, ABC's Sunday newsmaker show has been passed by Mr. Schieffer and "Face the Nation." NBC's Tim Russert and "Meet the Press" is far and away the leader, while "Face the Nation" and "This Week" duke it out for second place.
However, Mr. Stephanopoulos and executive producer Katherine O'Hearn, hired in 2005, have become more aggressive about booking political newsmakers, even if they have to travel internationally to do it.
For "This Week," the second quarter of 2006 was one of growth. According to data from Nielsen Media Research, "This Week" was up 28 percent year to year among the 25- to 54-year-old viewers who are the key news demographic and was up 16 percent in total viewers. Indeed, the second quarter was "This Week's" best since 2002 in total viewers and since 1999 in the demo. And the gap between "This Week" and "Face the Nation" was the smallest since 2001.
Had Mr. Stephanopoulos not stayed, ABC News would have been in a bit of a pickle because there is no one in its stable who could bring all the same qualities to the table, and because crucial midterm campaigns and elections have begun.
Mr. McManus would appear to confront the same challenge and more if he still wants to expand his political team to add expertise that enhances Ms. Couric's political credibility without making her seem as daughterly as she might if appearing next to Mr. Schieffer.
Even if Mr. McManus didn't poach Mr. Stephanopoulos, he forced ABC News to pay more than it otherwise would have to keep the political pundit.
In TV circles that's a bit of gamesmanship that makes the game fun even when the game plan doesn't pan out.
HBO, ABC Lead Creative Arts Emmys
Gandolfini Signs Exclusive HBO Deal


text:
Friday Five: March Madness, political correctness, and more
Welcome to the Friday Five! Every Friday we share our top five articles, stories, and quotes for you to read and share with your friends and church families.
As always, share any ideas for stories with us on Facebook and Twitter.
1. The NCAA must once and for all stop extorting North Carolina.
"The NCAA is supposed to be a non-profit. Their attacks against North Carolinians are beyond their sport's interest. Worse still, the NCAA is double-dribbling in hypocrisy. In addition to segregating collegiate sports by male and female, and moving games to states with similar nondiscrimination laws, the NCAA uses gender assigned at birth to prevent certain transgenders from playing on women's teams. (Read more)
2. House Republicans file HB 328 to request the IRS to review the nonprofit status of NCAA as a result of their substantial lobbying and economic boycotts
"An N.C. House Republican says the NCAA and ACC have "stepped out of bounds" by moving sports championships out of North Carolina over House Bill 2." (Read more)
The NCAA's response: "The National Collegiate Athletic Association (NCAA) has responded after North Carolina lawmakers filed a bill accusing the organization of "excessive lobbying" in the wake of House Bill 2." (Read more)
3. Texas Senate passes SB6, their HB2-inspired privacy bill. Other states also following NC's lead
"I will tell you as a woman, this is not a joke. This is about dressing rooms, lockers, showers and restrooms. This is about privacy and protection for all people," Kolkhorst said. (Read more)
4. VIDEO: Washington grandmother and florist loses her business by following her conscience
"For years, we have warned about the harmful effects non-discrimination ordinances have by forcing business owners like Washington florist Barronelle Stutzman to violate their religious convictions. Washington's harmful non-discrimination law has caused this 72-year old grandmother to lose her livelihood and possibly her home just for following her conscience, and this points to the virtue of having HB2 in place in NC, so that government can't coerce violations of conscience and take away freedom." (Read more)
5. Political correctness reaches new and ridiculous heights on college campus
"You know political correctness has reached new heights when a college English department forgoes standards in English grammar to appease cultural elites:
"The university's writing center recently released a guide on 'Inclusive Language & Gender Neutral Pronoun Use,' offering students two options: change all their sentences so that the subject is plural (for example, switch out 'a student' for 'students'), or simply replace the singular pronoun 'he' or 'she' with 'they'. To avoid confusion, the guide recommends students who choose to use 'they' as a singular third-person pronoun place a disclaimer at the top of their papers. " (Read more)

redpajama_set_name: RedPajamaCommonCrawl


Thank you so much for the car it is great. It past all the compliance and drives like a dream. My friend will enjoy it for years to come. Thank you so much for your amazing service and all the help you gave us along the way. I hope you continue to have great success in your business as you are so good to deal with.
All the best and I will be sure to tell my friends about your company and your cars.
A huge thank you to all at JPC Trade. I couldn't be happier with my Integra Type R. Even with my specific requirements we were able to find me a car that fit the bill perfectly.
The whole process was made simple from start to finish which made things very easy for my first time importing a car. A very enjoyable experience and I certainly would not hesitate to use JPC Trade again in the future.
I'd just like to thank you personally Murata for all your help in dealing with me and my car. I'm very happy with the level of customer service I received through out the entire process. Thank you very much!
This is my first car import by JPC TRADE. Ozawa was very kind and professional. Car arrived in very good condition, same as the auction sheet. No problem at all, easy to pass the complaints. JPC website is also very easy to use, much better than other agent website. I will import another car again soon from your guys.
Thank you so much to Keima and the Team at JPC trading to help us find our his and hers minis.
We researched about 6 month before deciding to embark on buying a car direct from the Japanese auctions. JPC was recommended to us by a friend that also purchased his vehicle from JPC and was very happy. Still, we compared about 5 direct importers before we selected JPC. Keima-San from JPC took the time to call us and consult us on our needs absolutely obligation free. Keima-San who lived in New Zealand spoke excellent English and was quick to grasp what we were looking for. Compared to others, he provided us with the best upfront information and installed confidence that our expectations will be met.
Once we started, Keima-san made the process a breeze, he took care of all bidding, paperwork and the booking for international shipping. After our purchase using the JPC customer interface I was able to track the cars every step of the way until arrival at the local compliance center. Both cars came in better condition as expected and thanks to Keima-sans excellent advice during car selection compliance was very simple.
Keima-San provided fantastic service, before, during and even after the car arrival and made it a wonderful experience.
We look forward to next time.


February will mark the 2 year anniversary of Halal Food Guru. Our goal when we first started posting was to make it easier for travelers to find and enjoy Halal restaurants in Florida. We wrote detailed reviews of these Halal restaurants so visitors would know what to expect and gain some local insight about the spots to make the most of their experience. We introduced TOP 5 Lists to make it easier to find what you crave, which was well received. Over the past 2 years, we've written over 50 posts about Halal restaurants in Orlando, Tampa Bay, Jacksonville, Miami and other great cities across North America. The feedback has been overwhelmingly positive.
We are thankful for all the support that you have shown us as we grow. We appreciate everyone who follows our website and/or social media pages including Facebook, Instagram and Twitter. It means a lot to us when you take the time to like and share our content.
Understand that this is a passion project as outlined in our About Page. We really want you to enjoy your trip to the Sunshine state and food is a big part of that. By highlighting Halal restaurants in the area, you can get more out of your day and vacation stay.
For this post, we would like to highlight some of our favorite TOP 5 lists from around Orlando. These are by far the most popular posts. In these TOP 5 posts, we pick a food category and find the 5 best Halal spots in the city that offer it up.
We don't know where the obsession with cheesesteaks in Orlando came from, but almost every Halal Fast Food spot offers their interpretation. And some are definitely to die for.
This was one of our first TOP 5 lists and continues to get a lot of attention. It's great to help plan your meals when you visit the theme park. Let me know if you want us to do other TOP 5 lists around other landmarks.
The New York street food dish made a big splash in Orlando in recent years and has become a staple in the Halal Fast Food scene around town. It's created a lot of White Sauce addicts!
There is lots to do in Orlando during the day, but once the sun goes down the tummy can grumble. Thankfully there are several Halal spots open late so you can eat great.
Another fan favorite are the Guru's Stories. In these stories we recount some of our foodie adventures which had an interesting or unexpected twist. A lot of our followers really got a kick out of our storytelling. So below is a link to the stories we have posted so far.
First time visiting Halal Guys in New York City. When you are on the hunt for the Original Halal Guys Street Cart on the streets of New York City, anything can and will happen.
I want Pineapple Juice NOW. Pregnancy can be tough and chaotic. But Pregnancy Cravings can really throw a monkey wrench into the mix.
Again, thank you so much for your support and encouragement. We couldn't grow to this point without you. We look forward to continuing to provide you with in-depth, honest reviews of Halal restaurants across the state. Please continue to like and share our posts with your family and friends, especially if they are planning a trip to Florida.


text:
class Responder
  attr_reader :params, :parsed_params, :channel
  def initialize(channel, params)
    @params = params
    @channel = channel
    parse_params
  end

  def parse_params
    @parsed_params = JSON.parse(params).with_indifferent_access
  end

  def intent_class_name
    channel.split(".").last.classify
  end

  def intent_class
    Object.const_get("Intent").const_get(intent_class_name)
  end

  def intent
    @intent ||= intent_class.new
  end

  def apply
    intent.apply(parsed_params)
  end

end

redpajama_set_name: RedPajamaGithub


text:
Wes Tacos
12 games reviewed
68.3 average score
68 median score
16.7% of games recommended
Are you Wes Tacos? If so, email [email protected] to claim this critic page.
Sort by: Page 1 of 2
5.5 / 10.0 - Smash Hit Plunder
Despite its inherent flaws and strange design choices, it's not by any means a bad game. It's just really not a very good one, either. You can have some fun with it, and it might even be a good way to introduce new people to VR. But at the end of the day Smash Hit Plunder feels more like a tech demo stretched out into a full game, without adding anything in to justify that decision. And that makes me more ill than any motion sickness ever could.
7 / 10.0 - Toki
Toki isn't a perfect game -- hell, I'm not sure it's even a great arcade game. But it really hit me in the nostalgia-feels. It's a fantastic remake from a technical perspective, even if it's totally lacking in extra features. If you like good old-fashioned arcade games, you can do a whole lot worse than this one. Just make sure you temper your expectations, as a lot has changed in the gaming landscape in the 30 years since its original release.
6 / 10.0 - Mutant Football League Dynasty Edition
Overall, it just feels like there are a lot of missed opportunities in Mutant Football League: Dynasty Edition. The gameplay certainly works, but there's really nothing all that spectacular about it. The violence is there, but it just feels tame even in comparison to a bloodless 20-year-old competitor. The game modes are all genre-standard, but there's really nothing to separate it from a normal sports game. The aesthetics are cool, but there really aren't any differences between players and races apart from cosmetics. It's punny, but not really all that funny. It's a game that's ultimately dragged down by too many buts to really stand out and make a name for itself.
6.5 / 10.0 - The Low Road
The Low Road isn't a bad game -- it's perfectly competent and serviceable in basically every way. It just doesn't take its great setup to anything more than a perfectly competent and serviceable game, which results in more of a big letdown than an objectively bad game.
6.5 / 10.0 - Operation Warcade
Operation Warcade isn't a terribly innovative game, but it's fun enough if you're just looking for a very basic arcade shooter. Just don't expect a definitive VR experience from it.
8 / 10.0 - Hopalong: The Badlands
There are a few glitches here and there and the final level is more frustrating than it probably needs to be, but when you're speeding by a gang of dynamite-wielding hooligans shooting shotguns and smacking them with a pickaxe, you'll forget all about the minor small annoyances. Hopalong: The Badlands is a fun and intuitive arcade shooter that lasts just long enough for you to feel like you've truly mastered its controls, but not too long to overstay its welcome.
9 / 10.0 - Wailing Heights
I thought the game looked kind of silly and fun when I first saw it, but I honestly didn't expect to enjoy it as much as I did. Sure, it was frustrating at times as I ran around the town with each character trying to trigger the one specific thing I needed to do to advance the story, and yeah there are some minor bugs that cause your characters to freeze or disappear behind some level architecture. But after the game was over, I had forgotten most of my previous complaints and found that I had legitimately enjoyed playing it. This has somehow become one of my favorite games, and I couldn't be happier about that.
5.5 / 10.0 - State of Anarchy: Master of Mayhem
State of Anarchy: Master of Mayhem is a decent twin-stick shooter that more or less works in function and fun, but has many setbacks that prevent it from being anything other than a middling affair. It's certainly not a bad way to get some light arcade action, but you'll hardly feel like you've mastered any mayhem by the end.
7 / 10.0 - Sausage Sports Club
Sausage Sports Club isn't a particularly girthy game, and the motion of its ocean is sometimes a bit choppy, but ultimately this is a unique and charming party game that'll leave your group of friends sloppily coming back for seconds.
6.5 / 10.0 - Tanzia
Tanzia's disparate parts are held together with the chewed gum of love put into the project which manages to do just enough to keep it all from falling apart. It definitely has limited appeal, but fans of that relative lack of modern polish we now attribute to sixth generation games will find a competent little action-RPG romp that stands tall with its niche PS2 counterparts.


Mecp2 null mice model Rett syndrome (RTT) a human neurological disorder affecting females after apparent normal pre- and peri-natal developmental periods. Neuroanatomical studies in cerebral cortex of RTT mouse models revealed delayed maturation of neuronal morphology and autonomous as well as non-cell autonomous reduction in dendritic complexity of postnatal cortical neurons. However, both morphometric parameters and high-resolution expression profile of cortical neurons at embryonic developmental stage have not yet been studied. Here we address these topics by using embryonic neuronal primary cultures from Mecp2 loss of function mouse model.
We show that embryonic primary cortical neurons of Mecp2 null mice display reduced neurite complexity possibly reflecting transcriptional changes. We used RNA-sequencing coupled with a bioinformatics comparative approach to identify and remove the contribution of variable and hard to quantify non-neuronal brain cells present in our in vitro cell cultures.
Our results support the need to investigate both Mecp2 morphological as well as molecular effect in neurons since prenatal developmental stage, long time before onset of Rett symptoms.
Mecp2 encodes a methylated DNA-binding protein and is the causative gene of Rett syndrome  (RTT, MIM #312750) a progressive neurodevelopmental disorder affecting 1:10,000 females worldwide . Patients are apparently healthy during the first 6–18 months of life, then the disorder causes mental retardation, deceleration of head growth, seizures, motor dysfunction, hand stereotypies together with many other disabling symptoms . Mecp2 is ubiquitously expressed with 2 splicing isoforms, Mecp2A and B , . In the brain these isoforms are developmentally and regionally regulated, with Mecp2B (also known as Mecp2α or Mecp2_e1) more abundant in adulthood. Both in humans and mice the delayed onset of RTT symptoms is in accordance with the postnatal increase of the Mecp2B protein .
Neuronal morphology plays a significant role in determining how neurons function and communicate. Specifically, it affects the ability of neurons to receive inputs from other cells and contributes to propagation of action potentials. The morphology of neurites also affects on how information is processed by the brain , . Like many disorders associated with mental retardation , neuropathology of RTT reveals an increase in neural density due to decrease in cell size, together with reduced neurite length and branching in post mortem brains, as well as in Mecp2-loss of function mutant mice (Mecp2-null). Delayed development of neuronal morphological features and maintenance of functional networks  in Mecp2-null mice suggest that MeCP2 impacts neuronal maturation .
More recent studies have shown that RTT is not exclusively a neuronal disease. Despite the low level of MeCP2 expression in astroglia, oligodendroglia and microglia , , conditionally Mecp2 loss in brain non-neuronal cells detrimentally influences dendritic integrity, synapses and protein expression in neurons , , .
Availability of many RTT mouse models  contributed to disentangle MeCP2 functions and demonstrate that many RTT symptoms can be reverted by postnatal re-activation of MeCP2 expression .
Initially described as a transcriptional silencer through the binding to methylated promoters and histone deacethylases recruitment, extensive studies in mouse models have depicted MeCP2 as a multifunctional protein able to activate gene expression, model nuclear architecture, and regulate alternative splicing and translation , . In particular, the precise role of MeCP2 as a transcription modulator is still under investigation. Expression profiling of whole brain homogenates from Mecp2 null mice and postmortem brains from RTT patients, revealed only subtle differences. Most productive efforts to identify MeCP2 regulated-targets took into account the problem of brain heterogeneity, in terms of regionality and cellularity, as a dilution factor which leads to underestimate the actual number of deregulated genes .
Studies conducted till this date, only evaluate post-natal transcriptional profiling as well as neuronal morphological parameters in RTT mouse models. Current hypothesis claims that key embryonic and perinatal developmental steps are not altered until presymptomatic stage (3–5 weeks in Mecp2 null male mice). A candidate gene approach study has shown that expression of important proteins for neuronal maturation (i.e. ID1 and ID2, inhibitor of differentiation) is impaired at embryonic day 15 (E15) in the cerebral cortex of Mecp2 null male mice, transiently (ID2) or with a long-lasting effect (ID1) over postnatal stages . Furthermore, MeCP2 has been shown to promote heterochromatin reorganization during neural differentiation of embryonic stem cells and in mouse primary cortical neurons , . In turn, chromatin organization changes are essential for neuronal development, as they impact on gene expression.
In light of these findings, in this study we investigate the effects of Mecp2 loss of functions in earlier developmental stages than those analysed so far. By performing morphological analyses of primary embryonic cortical neurons dissected from Mecp2 null mouse brains we show reduced neuritic arborization due to Mecp2 loss of function. We profiled embryonic wild type (WT) and Mecp2-/Y (null) gene expression using RNA-sequencing assays. The availability of cell-type specific datasets, allowed us to use bioinformatics methods to isolate neuron enriched deregulated genes affected by MeCP2 deficiency. Results from our study suggest a role of non-neuronal brain cells variability for reproducibility and validity of gene expression profiling readouts.
We examined primary cultures of neurons dissected from E15 cerebral cortices of Mecp2 null embryos (Mecp2 tm1.1Bird/J strain) to investigate whether changes in neuronal morphology already occur during early brain development of RTT mouse models. Since Mecp2 starts to be expressed in cortical layers at E14.5  mostly in early neuronal committed cells , we chose this stage to dissect cortices of Mecp2 null and WT littermates. We considered embryonic neuronal primary cultures a useful tool to address our topic, as considerable information has been gathered on their neuronal development and maturation .
Primary cultures are often heterogeneous and require characterization of their cell types (neurons versus non-neuronal cells, i.e. glia) and neuronal sub-types. These latter may be identified through neurotransmitter synthesized- and/or region-specific markers. The phenotype of our cells was investigated by immunofluorescence experiments performed with cell-type specific antibodies. Cells were labeled with anti- β-tubulin-III (Tuj1, neuronal marker), GABAergic neurons with anti-GABA (γ- aminobutyric acid) and astrocytes with anti-GFAP (glial fibrillary acidic protein). More than 90 % of cortical cells in WT and Mecp2 null mouse cultures were neurons, as judged by counting the Tuj1 positive cells counterstained with the nuclear marker DAPI (Fig. 1). The percentage of GABAergic positive neurons over Tuj1 positive cell was about 30–35 %, whereas GFAP-positive cells were rare (about 1 % of DAPI stained nuclei). An additional figure file shows this in more detail [see Additional file 1].
Subsequently to characterization, cortical cells were stained with Tuj1 antibody and morphometrically analyzed. We will refer to neurites because cells were in vitro cultured for only three days, not enough time to distinguish axons from dendrites. The length of the neurites was estimated by measuring the distance from the cell soma to the end of the primary neurites (neurites that originate directly from soma). Neurite branching was estimated by Sholl analysis, one of the most commonly used methods to quantify neuronal dendritic complexity . We find that the length of neurites does not differ between WT and Mecp2 null neuronal cultures (Fig. 2a). On the contrary, total branching and primary branching (i.e. the number of secondary neurite portion/fragment growing from the main neurite extension) are significantly lower in Mecp2 null neurons compared to age-matched WT control (Fig. 2b).
Primary cell cultures from WT and Mecp2 null mice were profiled by RNA-sequencing to obtain a high- resolution dataset and search for cortical genes possibly affected by MeCP2 loss during early mouse development.
As Mecp2 is an X-linked gene, we collected only male embryos to avoid the confounding effect of X chromosome inactivation typical of females on gene expression profiles. The inbred C57Bl/6 J strain was used to minimize genetic background influence. In order to minimize inter-individual variability, RNA isolated from cells of single embryos was pooled accordingly to Mecp2-genotypes in three samples/genotype (see Methods) and RNA from each sample was sequenced.
We obtained 30 million of reads per sample with quality scores higher than Q20, mapping on average 96 % of them to Mus musculus reference genome sequence (GRCm38). A hierarchical clusterization based on correlation was performed to check the quality of the samples (an additional figure file shows this in more detail [see Additional file 2]). One null and one WT samples did not cluster properly (data not shown), thus they were excluded from further analysis.
Total reads related to Mecp2 were comparable among WT samples (RPKM values: 10.21 and 9.22 for WT2 and WT3 respectively) suggesting similar levels of expression. In null samples, few Mecp2 specific reads (RPKM values: 0.90 and 0.84 for N2 and N3, respectively) mapped exclusively downstream to the deleted part of the gene. In addition, we found intergenic reads spanning the 3 kb genomic region from Mecp2 3′UTR to Irak1 locus (an additional figure file shows this in more detail [see Additional file 3, left panel]). The counts per million (CPM) of the reads mapping in this region was 5.5 and 0.04 for Mecp2-/Y and WT samples, respectively. The observed difference was found to be statistically significant after Welch Two Sample t-test (p <0.05) and transcription of this region was confirmed by RT-PCR with two different sets of primers ([see Additional file 3, right panel]).
As for the overall transcripts data set, by quantitative comparison and using a q-value (FDR) ≤0.05 we find 490 differentially expressed (DE) genes in E15 cortical neurons of null mice versus WT littermates, with 394 up-regulated (80 %) and 96 down-regulated (20 %) genes. DE genes cluster in 7 classes (Fig. 3) according to their expression profiles and their average fold-change (FC). Down-regulated genes show a mean FC of −0.93, whereas most part of up-regulated genes shows a mean FC of 1.22. Among up- regulated genes belonging to Cluster_3, we find GFAP (ENSMUSG00000020932) commonly referred to as an astrocyte marker. Even if GFAP has been already reported as a Mecp2-target in RTT patients  and in Mecp2-knock down rat model  this finding was not fully expected. Indeed GFAP was not found deregulated in primary astrocyte cultures prepared from postnatal day 1 cerebral cortex of the same null mouse strain  we used in this study. In fact, transient upregulation of GFAP in Mecp2-deficient cells seems to be female specific at least in amygdala and hypothalamus , whereas it increases in the hypothalamus of a male transgenic mouse overexpressing MeCP2 under its endogenous promoter .
Aldh1L1 (ENSMUSG00000030088), another gene mapping to the same FC based cluster of GFAP (cluster_3), is up- regulated as well in null samples versus controls. Aldh1L1 strongly labels many more astrocytes than GFAP . This finding, together with the fact that primary cells were 90 % composed of neurons (see previous results above), suggested us to verify whether the expression of other non-neuronal markers occurred across profiled samples. Microglia and oligodendrocyte markers (CD11B-ENSMUSG00000030786 and NG2/CC1- ENSMUSG00000015478/ENSMUSG00000005871, respectively) are indeed expressed in our in vitro cellular system, even if their expression ranged from very low to slightly low levels (not shown).
We next wanted to verify the hypothesis that some of the differentially expressed genes that we identified could be of non-neuronal origin. To do this we bioinformatically sorted the cells using available gene expression microarray data from the main central nervous system (CNS) neural cell types . We map the 490 differentially expressed genes resulted from RNA- sequencing across three different neural cell types. Figure 4 shows that 34 % of DE genes found in this study may be collected specifically under the astrocyte-signature (167/490), 9 % (45/490) under oligodendrocyte- cell type, while 14 % (71/490) of genes may be enriched in neuronal cells. We classify the remaining 196 DE genes (40 %) as unmapped. Most of these latter genes (168/196, see Additional file 4) were present on the GeneChip microarray (GSE9566) used in Cahoy's study  but probably they have not any cell type specificity. Moreover, as the expression of microglia marker-CD11B/Itgam was found among the 196 unmapped genes, we compared them to the recently reported microglia sensome , without finding yet any overlapping gene.
Firstly, we compared the list of the 167 genes enriched in astrocytes with those specifically deregulated in postnatal primary cultures of Mecp2-/Y astrocytes . Only 3 genes overlap with our astrocyte-enriched dataset: Gabrg1 (ENSMUSG00000001260), encoding GABA-A receptor gamma 1 concordantly deregulated in both datasets; Aldoc (ENSMUSG00000017390) and Fgfbp3 (ENSMUSG00000047632) that are discordant with results of Yasui ([see Additional file 5]). This finding, together with the fact that two astrocyte markers such as GFAP and Aldh1L1 (see above) have been found both up-regulated in our Mecp2-null derived primary cultures versus controls, raises the possibility that a larger amount of GFAP+ and Aldh1L1+ cells (even if rare) were originally present in the null compared to the WT cultures from which RNA for sequencing was isolated.
Among the 490 DE genes only 9 % are enriched in oligodendrocytes (Fig. 4). We find comparable expression of NG2, a marker of oligodendrocyte precursor cells, and CC1, a marker of mature oligodendrocytes. These markers are not differentially expressed with the respect to the genotype of samples (see Additional file 5).
According to Gene Ontology (GO) analysis (An additional composite figure file shows this in more detail [see Additional file 6A]) the most significantly represented biological processes are those related to ensheathment of neurons (especially myelination) and regulation of action potential (GO: 0042552 and 0019228, respectively with p-value ≤ 0.05). Given that the transcriptome dataset  profiles cells in postnatal mouse, GO outcomes would have been biased towards biological process more active in postnatal stages, as myelination does.
Nonetheless, GO together with Panther analysis (see Additional file 6B) give us interesting directions of investigation to be tested, as the most enriched annotations in this category of genes are related to protein families involved in oligodendrocytes differentiation, such as Olig 1 and 2 , CD82  and CD9 , or to lipid synthesis ([see Additional file 6A]), such as ELOVL7  whose expression increases in null cells here sequenced ([see Additional file 5]). To be noticed, aberrant oligodendrocytes maturation could affect neuronal-non neuronal cells interactions  as reported for astroglia. Furthermore, among the 45 deregulated genes within oligodendrocyte- cell type no matches have been found in the cerebral cortex dataset of developmentally regulated genes  suggesting that their expression level might be constant during brain development.
Functional annotation of the 196 DE genes not mapped to neurons or glial cells shows that they are significantly enriched in biological process like the Tumor necrosis factor (TNF)-mediated signaling pathway (An additional figure file shows this in more detail [see Additional file 7). In response to numerous stimuli, TNF-pathway triggers downstream effects related to cell viability, gene transcription, ion homeostasis and synaptic plasticity. This pathway as well as activated microglia are key mediators of neuroinflammation and may contribute to neuronal dysfunction.
This functional outcome may have important molecular implications on the neuronal morphological phenotype here described, as it has been shown that postnatal Mecp2 null mouse microglia detrimentally affects dendrites and synapses by releasing an excess of glutamate .
In addition, as perturbation of embryonic microglial activity impairs normal assembling of cortical networks  there could be a causal relationship between MeCP2-deficient microglia and forebrain connectivity even during embryonic development.
The microglial marker CD11B indeed maps to this category but it results to be up-regulated in our Mecp2 null cell cultures versus WT control, possibly indicating a different distribution of CD11B+ cells across profiled samples, as another report indicates that CD11B is not altered by Mecp2 deficiency .
Of the 71 genes assigned to neuronal cells, forty-three genes (60 %) are more expressed in Mecp2 null cells than in WT while the remaining twenty-seven genes (40 %) are down-regulated ([see Additional file 5]).
Before further characterizing these genes we checked whether neuronal and GABAergic markers were equally expressed across samples. The overall number of neurons does not differ between WT and null samples, as previously shown by Tuj1-immunofluorescence staining (see above) and then confirmed at transcriptional level by RNA-sequencing (β-tubulin-III- ENSMUSG00000072235 RPKM values in WT samples are 1202.42, and 1081.06, respectively; in null samples 1043.82 and 1114.27, respectively). Similarly, there are no differences in expression level of: a) GABAergic marker-transcripts, such as GABA synthesis enzyme GAD2 (ENSMUSG00000026787, RPKM values in WT samples are 8.30 and 6.46, respectively; in null samples 6.51 and 6.58, respectively); b) GAD1 (ENSMUSG00000070880); c) genes encoding the GABA transporters, such as Slc32a1 and Slc6a1 together with Dlx1 and Arx  ([see Additional file 5]).
Assuming homogeneity of samples, the list of DE genes carrying a neuronal transcriptional signature was then analyzed, to search for functional categories more enriched and potentially targeted by Mecp2 loss. Functional clustering (Fig. 5) shows 7 significantly represented clusters (p-value < 0.05), with three molecular function terms (clusters A, C and D) and four biological process annotations (clusters B, E-G). Interestingly, the major functional themes assigned are specifically related to neuron specific features, such as neurogenesis and cell differentiation (cluster E) together with neuronal activities. Indeed, ion binding (cluster A), transport (cluster B) and channel activity (Cluster C, D and F) altogether sustain synaptic transmission. Among genes listed in these functional clusters, some contribute to shape neuronal morphology, such as Mef2C and Tiam1 (also identified in ). Another interesting gene found in these clusters is GDA encoding guanine deaminase, a protein involved in microtubule assembling  and regulation of dendrites number . To further validate the GO and functional annotation results, we carried out enrichments analysis of each GO and functional terms separately for the given list of neuronal specific genes. We obtained enriched GO and functional terms along with multiple test corrected p-values (An additional table file shows this in more detail [see Additional file 8]). Together with DAVID annotation, we carried out GO annotation analysis through AmiGO 2, obtaining significant GO terms using Bonferroni correction method (p-value < 0.05). The results (see Additional file 8) confirm the biological inference for the neuronal specific genes obtained by DAVID analysis.
To understand whether the neuronal genes mis-regulation we found at embryonic stage is conserved throughout development, we compared our data with a number of gene expression studies profiling various brain regions of postnatal Mecp2 mutant brain. To avoid any mouse background confounding effects, we focused our interest in regions dissected from the same Mecp2 null mouse strain ,  analyzed in this study. We find that the highest rate of sharing is achieved with hypothalamic genes: twenty-seven out of seventy genes (excluding Mecp2) are differentially expressed in postnatal hypothalamus of Mecp2 null mice  with 12 genes (44 %) deregulated in the same direction ([see Additional file 5]). Only two genes are concordantly shared with cerebellum of Mecp2 null mice, with Pls3 (ENSMUSG00000016382) gene also present in the hypothalamus list ([see Additional file 5]). The pairwise comparison of the 71 neuronal DE genes with those published by Zhao and colleagues  in symptomatic P60 (postnatal day 60) Mecp2 null mice (Bird strain) revealed only 4 genes in common (see Additional file 5). Nonetheless, all neuron related GO terms above mentioned are also shared between the two datasets, uncovering specific functions of MeCP2 regulated genes during development.
Screening the cerebral cortex dataset of healthy mice  we find 30 genes ([see Additional file 5]) merging with our neuronal enriched dataset. Notably, 90 % of them become more highly expressed in adult cortex during mouse development, including 7/8 deregulated genes concordant with hypothalamic findings.
Finally, we find that 50 % of neuronal enriched up-regulated genes (22/44, data not shown) are long genes (around or more than 100 kb) as already reported for MeCP2-repressed genes , .
Neuroanatomical studies in cerebral cortex of Mecp2 null mice showed delayed maturation of neuronal morphology and autonomous as well as non-cell autonomous reduction of dendritic complexity of postnatal cortical neurons. In addition, abnormal cell body size and density of both glutamatergic – and GABAergic neurons  has been reported for a RTT mouse model  as well as for inhibitory neurons differentiated from human embryonic stem cells lacking Mecp2 .
In this study, we describe effects of Mecp2 loss of function both on the morphology and transcription profile of the embryonic neocortical neurons. Interestingly, we find a reduced neurite branching in Mecp2 null neurons, whereas neurites length is not affected. Similar results have been recently described in inhibitory neurons differentiated from human embryonic stem cells lacking Mecp2 . In addition, we identify a neuronal subset of embryonic deregulated genes that are highly expressed in mouse adult cortex, possibly indicating that some subtle embryonic defects will be functionally amplified later in postnatal life when clinical RTT symptoms become evident. The findings in our study further support the hypothesis that Mecp2 has a role in structural plasticity of cortical neurons and highlight the effects on the transcription of a subset of neuronal genes. It has been recently reported that Mecp2 deficient mice display regional- and age-dependent variations in the GABA pathway and, to a lesser extent, in the glutamate pathway . Focusing on functional annotations significantly enriched in neuronal cells here profiled, we achieved additional as well as interesting results: MeCP2 deficiency should impair the same biological process even across different brain areas and developmental stages, as suggested by the overlapping of Gene Ontology terms with those described for striatal deregulated genes of postnatal Mecp2 null mice .
However, it would be interesting to measure in our cellular system the impact of non-neuronal cells on the transcription of neuronal genes. Indeed, the deregulated genes due to the Mecp2 loss only in the striatal GABAergic neurons partially overlap with those impaired by a constitutive Mecp2 loss .
Our bioinformatics approach on the list of differentially expressed genes in primary cultures from Mecp2 null and WT cortices has another important outcome: on one hand allows to focus on genes with a neuron transcriptional signature, while on the other identifies non-neuronal enriched genes contribution to the list. Overall findings from non-neuronal cortical cells may suffer from inter- individual variability and chance to survive in serum-free culturing medium. Prior RNA-sequencing, pooling of samples (merely based on genotype and gender of embryos) may have masked such variability, giving rise only to partial conclusions of Mecp2 loss of function effects. The simultaneous presence in our cultures of different neural populations allowed us to preserve cell-cell interactions between neurons and non-neuronal cells. This is a fundamental aspect in molecular and morphological studies of Mecp2 null cells, as neuron-glia and -microglia interactions have emerged as a crucial point to control RTT progression and for some of them to reverse symptoms.
There is an extensive reorganization of mouse transcriptome between embryonic and postnatal developmental stages , . Moreover, gene expression programs are highly divergent across cell types . Likely, not overlapping results of downstream effectors of Mecp2 deficiency can be also affected by the specific developmental window of profiling. In addition, it has been shown that sorting methods to profile specific cell types can themselves produce in some cases not comparable results . Despite this, a small subset of deregulated neuronal genes here identified has already been described in a previous study  at a postnatal stage and in a different brain region (hypothalamus). We speculate that Mecp2 loss may impact transcripts abundance of those genes in a space-temporal independent ways, starting from mid-gestation period. Interestingly, at E15 when those genes are deregulated in the cortex, MeCP2 protein is not detectable in hypothalamus .
Most part of neuron-enriched deregulated genes here found are up-regulated, whereas in previous studies most part of Mecp2 deregulated genes are repressed. This discrepancy may be likely linked to different developmental stages analysed but also to the technology utilized, given that we were able to measure even very low expressed genes (see Fig. 3). However the rate of up-regulated genes here found is consistent with that reported by Sugino  based on cell type specific approach. Moreover, concordantly to Sugino et al.  and Gabel and et al.  we also find that genes up-regulated by Mecp2 loss are biased toward long genes.
In addition, RNA-sequencing permitted us to confirm that Irak1 deregulation is causally linked to genetic structural manipulation of mouse Mecp2 locus, rather than to functional loss of Mecp2. Indeed Irak1 levels specifically increase in the Bird strain of Mecp2 null mice lacking exons 3 and 4 , whereas in the Jaenisch strain is normally expressed . In all brain regions to date profiled , , – and in this study, Irak1 is always up-regulated. Our mapping of reads in the intergenic region between Mecp2 and Irak1 in null samples may be due to spurious transcription occurring in the absence of terminal coding part of Mecp2 gene. It is thus reasonable to suppose that at least a subset of differentially regulated genes found in this study and by others should be considered as Irak1- dependent downstream effects rather than Mecp2 targets. Among downstream effectors of Irak1 signalling pathway there is NF-Kb, a transcription factor even acting as a modulator of neurite outgrowth . Moreover, the virtual cell sorting we performed assigned Irak1 to cells resembling activated microglia, characterized by TNF-mediated signal pathway enrichment. This finding supports an already proposed hypothesis of a constitutive activity of signalling pathways mediating the TNF- action, as Mecp2 null microglia releases significantly less TNF-alpha under specific stimulation . Remarkably, our findings motivate further studies focused on genes involved in shaping cell morphology, whose mis-expression can lead to altered formation, pruning and maintenance of neurites and synapses. Mef2c and Tiam1, already described as Mecp2 target genes, may be good candidates. Mef2c negatively regulates synapses number and function  and is up-regulated by Mecp2 loss (this study and . Tiam1, essential for Rac1 activation and neurite outgrowth of cortical neurons , is slightly down-regulated in the cellular system we analyzed. As Tiam1 manifests the opposite response to Mecp2 loss in postnatal hypothalamus , we can suppose that Mecp2 may exert regional and/or stage specific regulation on specific genes. On the contrary, the up-regulation of the GDA gene may be in accordance with a model of compensatory strategy of neuron to recover branching defects , whereas the up-regulation of thyrotropin releasing hormone (TRH)-degrading enzyme would have a direct impact on action potential shaping of cortical neurons.
To our knowledge dendritic complexity of embryonic cortical neurons, as well as the embryonic high- resolution expression profile was never examined in Mecp2 null mice. Using a very sensitive technology we were able to profile even genes transcribed at low level, showing that our close to pure primary neuronal cultures derived from embryonic cortices may in fact contain low and poorly detectable amount of non neuronal cells. The virtual a posteriori cell sorting here described allowed also to focus specifically on differentially expressed genes of neuronal origin. We are confident that the cellular system here used could be further investigated, providing a fine characterization of neuronal subpopulation together with non-neuronal cells.
We thus suggest that abnormal morphology and altered expression profile of neurons may be very early phenotypes of RTT mouse models. Furthermore, subtle embryonic defects will be functionally amplified later in postnatal life, when overt RTT phenotypes will be established.
Female Mecp2 heterozygous females  were purchased from Jackson Laboratories (strain name: B6.129P2(C) − Mecp2tm1.1Bird/J, stock number 003890). Mecp2 mutant hemizygous males (Mecp2 -/Y, also indicated as Mecp2 null) and wild type (WT) littermates used as controls were obtained by mating heterozygous females with WT males (C57BL6/J background) for at least 12 generations. The national or institutional guidelines were used for the care and use of animals, and approval for the experiments were obtained from the ethical committees of the Italian Ministry of Health and the UK Home Office.
Genomic DNA was extracted from tail tips of embryos and immediately genotyped by polymerase chain reaction to determine Mecp2 deletion (according to ) and mice gender. The gender was determined using primers mSRY.F 5′-CCCAGCAGAATCCCAGCATGC-3′ and mSRYl.R 5′-TCCTGTCCCACTGCAGAAGGT-3′ specific for the Y chromosome-linked Sry gene.
Cortices from each embryo were separately dissected from those of the other littermates.
Dissociated cells were prepared by dissecting out cortices from individual embryos at embryonic day 15 (E15). The day of insemination (i.e. the appearance of a vaginal plug) is designated as embryonic day 0 (E0).
Cortices were dissected under a stereoscope in sterile conditions and placed in phosphate buffered saline (PBS) without calcium and magnesium, supplemented with 33 mM glucose. Cells were then cultured as previously described . Briefly, the dissected areas were enzymatically dissociated by incubation for 20 min (min) at 37 °C in a papain solution (Warthington, 20 U/ml, Milan, Italy) in Earle's balance salts containing 1 mM EDTA (Sigma-Aldrich, Milan, Italy), 1 mM cysteine (Sigma-Aldrich) and 0,01 % pancreatic DNAse (Sigma-Aldrich). After addition of 1 mg/ml of bovine serum albumin (Sigma-Aldrich) and 1 mg/ml ovomucoid (Sigma/Aldrich) the cells suspensions were centrifuged 5 min at 800 rpm, resuspended in plating medium and counted . For the viable cell count, cell suspension was diluted 1:1 with 0,1 % trypan blue dye (Sigma Aldrich) and loaded into a disposable cell counting chamber-slide. Cell concentration was determined on the basis of the total cell count, the dilution factor and the trypan blue dye exclusion.
Dissociated cells were plated at a density of 1 × 105/cm2 in 2 cm2 cell cultures dishes (Corning) coated with 15 μg/ml of poly-D-lysine dissolved in water (Sigma-Aldrich).
Cultures were grown in serum-free Neurobasal medium (Life technologies, Milan, Italy), supplemented with B27 (Life technologies), 2 mM L-glutamine (Sigma-Aldrich), penicillin (50U/ml, Sigma-Aldrich) and streptomycin (50 μg/ml, Sigma-Aldrich). Cells were maintained for 3 days in vitro (DIV) at 37 °C in a humidified incubator in presence of 5 % CO2, before experimental manipulation. For each experimental point, cultures were prepared at least in independent triplicates, and were repeated using distinct culturing sessions.
Only primary cultures deriving from male embryos (see above) were then selected for subsequent analyses.
For immunocytochemical analyses cultured cells were fixed in 4 % paraformaldehyde in PBS, for 30 min at room temperature (RT), washed three times in PBS, and then permeabilized for 20 min in PBS containing 0,1 % Triton-X-100 and 10 % normal goat serum (NGS). Cells were treated with blocking solution [10 % NGS, 0,1 % bovine serum albumine (BSA) in PBS] at RT for 1 h and incubated with the primary antibody in antibody solution (PBS containing 0,1 % BSA) overnight at 4 °C. The following antibodies were used at the indicated dilutions: monoclonal antibody against neuron specific class III β-tubulin (Tuj1, Covance, Milan, Italy) 1:500, polyclonal rabbit antibody anti-γ-Aminobutyric acid (GABA, Sigma, 1:500) and polyclonal rabbit antibody against Glial Fibrillary Acidic Protein (GFAP, Dako, Z 0334) 1:450.
Cells were washed 3 times in PBS, and then incubated for 2 h at RT with fluorescent-labeled secondary antibodies (Alexa Fluor Goat anti-rabbit, and Alexa Fluor Goat anti-mouse, Life Technologies) diluted 1:400 in antibody solution.
Cells were then counterstained with DAPI (nuclear stain, 1:1000) for 10 min, washed with PBS and mounted with oil mounting solution (Mowiol). As negative controls, some cells were processed as described above, but without primary antibody.
For morphological analyses, cells fixed and permeabilized as above, were incubated with the monoclonal antibody against neuron specific class III β-tubulin (anti-Tuj1, 1:750, Covance) and with the fluorescent-labeled secondary antibody, both diluted in PBS containing 10 % NGS, for 2 h at RT. After Tuj1 staining, cell-culture slides were analyzed with a Leica microscope (Leica DM6000B) using the software Leica Application Suite (LAS AF). Images were obtained by using a 20x objective and captured with a HAMAMATSU Digital Camera C10600 ORCA R2. Measurements of neurites length have been performed by the image-processing software Image J. Images were pre-processed to optimize illumination and contrast. The length of the neurites was estimated by measuring the length of a line manually drawn from the soma to the end of the primary neurites (neurites that originate directly from soma) using the "Measure" function of the software.
The primary branching was measured by the Image J plugin "Sholl analysis". Its internal algorithm creates a series of concentric circles around the soma of the neuron and counts the number of neurites crossing these circles. Only clearly visible cells were analysed to prevent inaccurate scoring. For morphological analyses, a total of 15 fields for each cell-culture condition were used from at least three independent culture wells.
A total of 300 neurites/well were traced from Tuj1 positive neurons to measure their length. The analyses were carried out "blind" to avoid any subjective influences during measurements.
Total RNA was prepared from neuronal primary cultures using TRIZOL Reagent (Invitrogen) with addition of 150 μg of RNAase-free glycogen in order to maximize yield. RNA was then treated with RNase-free DNase (Ambion, DNA-free kit).
To minimize inter-individual variability, equal amount of quality checked RNAs from three individual embryos were mixed to constitute a pooled sample. With this procedure we obtained 3 samples/genotype with 1:1 proportion between RNAs derived from each Mecp2 null embryo (males) and those isolated from WT littermates. Samples were labeled as WT1-3 or N1-3.
RNA libraries were constructed using the TruSeq Stranded mRNA Sample preparation kit (Illumina), and single-strand sequenced on Illumina Hiseq2500 platform at the Istituto di Genomica Applicata (Udine, Italy). The first purification step isolates the poly-A containing mRNA molecules using poly-T oligo attached magnetic beads. During the second elution the poly-A RNA is also fragmented and primed for cDNA synthesis.
To verify the changes in expression of the intergenic region between Mecp2 and Irak1 loci, RT-PCR was performed on independent samples from additional animals. For each sample 1 μg of total RNA prepared from neuronal primary cultures was digested with DNase (Ambion, DNA-free kit) and reverse-transcribed by random-priming using SuperScript II first strand kit (Invitrogen). The following mouse primers were used for PCR analysis: inter5F 5′-GTCCCTAATGGGAGAACCGA-3′ and inter2R 5′-AGGTGAGTGGCAATGGCTAA-3′; inter3F 5′-CCCAGCTCCTCTCTTGAACAC-3′ and inter3R 5′-CTTTCAGCTGGCCAAACAAGG-3′. As loading control, GAPDH primers have been used: gapdhup 5′- TGCACCACCAACTGCTTAGC-3′ and gapdhlw 5′-TCTTCTGGGTGGCAGTGATG-3′.
Raw reads were processed with Trimmomatic  in order to remove low quality nucleotides and adapters. The minimum phred quality score for bases was set to 35 and only reads with a minimum length of 25 bp were retained after trimming. The high quality reads were aligned against the Mus musculus reference genome sequence (GRCm38) with TopHat  version 2.0.9. The resulting alignment files were used as input for HTSeq-count  version 0.5.4p2 together with the GRCm38 annotation file to calculate gene expression values (read counts). Genes with very low read counts or those that were too variable among the replicates were removed with HTSFilter . Differential expression analysis was performed in R with the package TCC  in combination with edgeR . The coordinates of the intergenic region between Irak1 (ENSMUSG00000031392) and Mecp2 (ENSMUSG00000031393) and the number of the mapped reads in each sample were obtained with bedtools (v2.17.0).
Expression profiles of the differentially expressed genes were used to perform a K-mean clustering with MeV  and 7 clusters were identified by using Pearson Correlation as distance metric.
Gene Ontology (GO) and functional annotation enrichment analysis was carried out using Database for Annotation, Visualization and Integrated Discovery (DAVID) bioinformatics resources 6.7 (http://david.abcc.ncifcrf.gov/) maintained by National Institute of Allergy and Infectious diseases (NIAID), NIH. We used both DAVID web interface as well as custom made Python script implementing DAVID-web services in our in-house built Transcriptator software .
Transcriptator helps in determining automatic and reproducible GO and functional annotation results for the differentially expressed list of genes derived from transcriptomic analysis of data. Currently, DAVID provides annotation for 40 different categories. In our computational pipeline we includes GO terms, protein-protein interactions, protein functional domains, bio-pathways, sequence general features and gene functional summaries. Using DAVID web application, we exploited functional annotation enrichment as well as functional annotation clustering tools to obtain clusters of differentially expressed genes based on common functionalities. For the GO and functional annotation enrichment, we defined a stringent Ease score (p- value) ≤0.05 and Count ≥5, which is basically a modified Fischer exact test p-value to examine more conservatively the enrichment situation.
For neuronal specific genes, we carried out functional annotation clustering based on the algorithm in DAVID, which hypothesizes that similar annotations should have similar gene members. It integrates the Kappa statistics to measure the degree of common genes between two annotations and fuzzy heuristic clustering to classify groups of similar annotations according to kappa values. In easier terms, the higher the number of shared annotations terms, the greater the probability that genes will be grouped together , .
For the functional annotation clustering implemented algorithm and utilized in DAVID web application we selected a stringent cut-off Ease score ≤0.05 and an enrichment score value ≥ 1. We also carried out GO annotation analysis through AmiGO 2 (version: 2.2.0 amigo2b) with GO database release 2015-06-06. Jvenn software was used to create Venn diagrams .
The datasets supporting the results of this article are included within the article as Additional file 5.
Publication costs for this article were funded by Associazione Italiana Rett project "Studio di due approcci farmacologici per ripristinare i difetti molecolari, morfofunzionali e neurocomportamentali in modelli di laboratorio preclinici sperimentali della Sindrome di Rett" to MDE.
The authors acknowledge dr. V. Colonna for critical reading of the manuscript.
MDE gratefully acknowledges Associazione Italiana Rett for supporting project "Studio di due approcci farmacologici per ripristinare i difetti molecolari, morfofunzionali e neurocomportamentali in modelli di laboratorio preclinici sperimentali della Sindrome di Rett (2014)".
This work has been partially funded by MSC-ITN "DISCHROM" Project 238242 (to MDE); by INTEROMICS Flagship Project, PON02-00612-3461281 and PON02-00619-3470457. MRG work is conducted at National Research University Higher School of Economics and supported by RSF grant 14-41-00039.
MV, LS, FS, FM and MM contributed mice sampling and carried out the molecular and cellular analysis. KPT, RAC and WS performed bioinformatics and statistical analyses. MV, LS, RAC and KPT contributed to write the manuscript. MV, CPC, MRG and MDE conceived the study. All authors read and approved the final manuscript.


During the 2001 Legislative session, the Utah State Legislature eliminated funding for all public art projects for the fiscal year 2002, terminating a public program that has benefited the community for over fifteen years.
This means that citizens of the State of Utah from all walks of life and every part of the State will not have the opportunity to experience art in public spaces for the new or remodeled State buildings that were approved for construction this year. Buildings like the new center for the Shakespearean Festival, the Museum of Natural History and the performing arts center at Snow College, among others, did not receive funding for public art projects.
Communities have lost an opportunity to bring in artists from all over Utah and the United States to create art specifically for them and the institution or agency that serve them. The Public Art Program and the Utah Arts Council mourn the loss of this opportunity to help build the communities of Utah economically, educationally and culturally.
The cultural loss is incalculable. The loss to artists and small business is enormous. The loss to community efforts in establishing a sense of place and pride could never be measured. It is known that businesses looking to locate in a new community often look for the visible commitment to culture and pride of place that public art demonstrates.
Public Art is the very embodiment of art for everyone. It is a direct contradiction to accusations of elitism so often cited by opponents to the public funding of art.
Since the inception of Utah's Public Art Program by the Utah State Legislature in 1985, over 200 artworks have been installed in 63 sites throughout Utah. Our public art collection has a wide variety of media from stone gardens, benches and fountains to 19' paintings depicting the majesty of Utah's landscape.
All of these works are in, on, or around the state buildings they were commissioned for from St. George to Logan.
The process of commissioning public art is for and by the community. Every project has a unique committee. The architect, the Division of Facilities Construction and Management, the Utah Arts Council, and the state agency being housed in the facility and the community are all represented.
The committee reviews every artist's proposal and awards the commission based on the aesthetic value, qualifications of the artist and how appropriate the proposal is for the agency and the community. This process insures the art is not "plopped" into a community but is created specifically for and with sensitivity to the community's history, culture and people.
The Utah State Legislature's Capital Facilities and Administrative Services Committee is the body through which all funding for State facilities is approved or denied. This committee will be revisiting the issue of funding public artwork in the next legislative session. Please join us in contacting your legislative representatives to explain the importance of art in public spaces for your community.
If you have any questions about Utah's Public Art Program or would like some literature about the program please call or email Jim Glenn at 801-533-3585 or jglenn@ucc.state.ut.us or Fletcher Booth at 801-533-3586 or fbooth@ucc.state.ut.us.
The Utah Arts Council web site at www.arts.utah.org can give you more information about all of the programs offered through the Utah Arts Council. The Public Art "Commissions" page lists public art calls for entries from both regional and national projects.
"Ascent Totem" 1991 (Painted aluminum 25′ x 12′ x 4′) located at the College of Eastern Utah, Price UT. Images copyright Utah Arts Council.
We believe we have benefited greatly from the Public Art Program. Our campus has several sculptures as a result of the program that add beauty and interest to the campus. We bring visitors to our campus whenever possible. I'm sure that some individuals are surprised to see art at a technical school – but art enhances the soul regardless of your career choice. The comments we receive from the public are very positive, especially with regard to the most recent sculpture. It is truly quite impressive. I found the process to be fair and appropriate. The staff were great – gave us a lot of support and unbiased input, and did not try to influence the process. We had significant community input but appreciated that we also had a somewhat greater say in what was selected, for after all the sculptures are located on our campus – we look at them every day. I am truly sorry that funding has been cut. I understand the economic realities – when times are tight, we need to preserve the basics. It is unfortunate that some of the first items to go are those that elevate the human spirit – and in these recent times, we sorely are in need of that.
When, in February 2001, the Utah Legislature's Executive Appropriations Committee proposed to eliminate funding for art in public buildings, Representative Ralph Becker attempted to save the Public Art Program by amending the bill to allow interest earnings from tax funds for construction projects to pay for the one percent for arts program. After committee discussion, however, Rep. Becker's motion was withdrawn.
Representative Becker sent 15 BYTES the following in regards to the upcoming 2002 Legislative session:"Art deserves support from our State Legislature, and should be viewed as an essential part of our state buildings. Public art creates an atmosphere, heritage, enjoyment, and sense of place. Public art creates a lasting statement that we, in Utah, value art as an integral part of our community.
We also provide support for artists in Utah. The art element in our State buildings should not be viewed as extraneous. In the 2001 Legislative Session, the 1% for the arts was eliminated in many state buildings. I objected to the elimination, and tried unsuccessfully to restore the cuts. I believe we were creating a terrible precedent that I hope won't be repeated.
"Untitled Trees" by Kenny Davis appears in the Ogden Courthouse courtesy of the Utah Public Art Program. The work was done in 1995, measures 12′ x 8′ and is oil on canvas. Copyright Utah Arts Council.
In Ogden we believe that public art adds value to the community, so five years ago we passed our own Percent for Art Ordinance to launch our local program. The State Percent for Art Program has also provided art pieces that enhance the State owned buildings in our community, notably the Courts Buildings, the School for the Deaf and Blind, and Weber State University. Donations of additional sculptural pieces have resulted from the stimulation of our Public Art Programs.
I strongly support the reinstatement of the State's Program to apply 1% of a building's cost for art in public buildings. This program has made a tremendously positive impact on the visual aesthetic in communities all across the State.
I was fortunate to be invited to join the Selection Committee for the piece installed recently at the Ogden-Weber Applied Technology College. The process worked very well with an introductory pre-proposal meeting, a meeting to review 15 written proposals and select semifinalists, and a final meeting to select the finalist from 3 who presented models with more specifically designed proposals. The Selection Committee was made up of client representatives and arts professionals from the community, and each participated thoughtfully throughout the process.
Jim Glenn and Fletcher Booth from the Utah Arts Council's staff prepared informative documents and agendas for each meeting. After careful consideration, the Committee recommended that Robert Sindorf create his, "Dance of Creation" for the site, just to the north of the Manufacturing Technologies Building. Mr. Sindorf is an architect who fully understood the appropriate scale for this piece and who used tooling techniques in creating this work that reflected activities in this training facility. It is a visually stunning addition to the inventory of public art in Ogden. The selection of this work adds variety that further enables our citizens and visitors to understand and appreciate art. Those in the community, especially at the ATC, have really embraced this work and are thrilled to have it installed in Ogden.


text: export * from './entry-editor-dialog.component';


text:
Optimizing Immersion
Targeted Input and the Competence Gap
Posted by Terry Waltz on August 16th, 2016
I am glad to see that Stephen Krashen has reconsidered the wording which previously stated that TPRS becomes AL-M when the input is targeted.
However, removing the inflammatory language hasn't changed the basic idea being put forth. Krashen says he is simply describing two types of input (targeted, as in coming from a curriculum, and untargeted, as in arising naturally from interaction with students), but there are significant side comments included that indicate that the "right" kind of input, in his opinion, seems to be "untargeted". TPRS/CI teachers using "targeted" input are dismissed as grammar-pattern pounding, uncreative types who are simply in there to repeat things over and over hoping for immediate acquired output.
My personal opinion is that either type of input is just dandy, as long as it's comprehensible and there's enough of it. However, we teach in the real world. The vast majority of teachers are hired as public school teachers to implement a curriculum. That curriculum document is the set of language our students are to have worked with by the end of the year. As TPRS teachers, we do not subscribe to the concept of "Today is September 24, so they 'know' the preterite now". TPRS in practice has evolved far away from the workshop ideal of 30-minutes-and-they-have-the-language. Demos work that way. Classes don't.
But in a very damaging way, since Krashen is viewed as such an impeccable source, some of the arguments put forth to undermine targeted input are simply ill-founded. There are mischaracterizations of TPRS/CI being made to make a point, and all the argument flows from these basic misunderstandings of what TPRS/CI is really about in the classroom.
From Krashen's description of targeted input:
"Targeting 1 (T1):
3. The source of the rules to be targeted is external, from a syllabus made by others. Our job is to find a story or interesting activity that will include lots of comprehensibe/interesting repetitions of these items. (This is why we get questions such as "do you know a story which I can use for teaching the conditional?"). "
That is not "our job" as TPRS teachers. With all due respect to a fine theoretician, that is not at all what a reasonably-skilled classroom teacher using TPRS does, and anyone who knows TPRS from inside a classroom will be well aware of that. If a teacher believes that is "the job" as a TPRS teacher, that teacher has not been adequately trained in TPRS, and what the teacher is doing is not really TPRS.
This is where Krashen's description becomes over-simplified and extreme. Sure, there are un(der)trained teachers trying to use TPRS or CI, who will think: I will try to "find a story" because I think it's about "telling stories" or I can't do story-asking in the first place, only easier variants that come with a pre-made story (soooo popular these days since they are quicker to implement and require less training and practice), and — here's the crux of the matter — I have never been taught how to reliably elicit a story from students. There are steps that can easily be used to teach teachers how to do this and we do it quite successfully all the time. But there is no logical relationship between pre-planned language input and the kind of teaching described here. It doesn't make sense to condemn the input type for the fault of incomplete training.
"Thus, Targeting 1 is a way of "contextualizing grammar," defined here as beginning with a target grammar rule and finding a context that will help make it comprehensible."
No, "targeting 1″ is a way of teaching the language one is contracted to teach, because the community (school board, etc.) has decided that is the language it wants the students to work with at that particular level or class.
"Targeting 1″ is a way of teaching the language that will enable students to adequately take on some other task, such as reading a particular book in a language where there are no cognates and the script is not phonetic. The concept of untargeted input soon breaks down when you deal with a language that does not have dozens of substantial pieces of level-appropriate writing available which can be read by students without any sort of problem because "hearing the language is seeing it".
Equating "targeted input" with "teaching grammar rules" is another leap that is not well-founded in the practice of TPRS/CI by trained teachers. A teacher who knows what s/he is doing is hardly going to start the day by saying "Today I make the imperfect tense comprehensible". It seems to me that these are all very extreme examples taken from teachers who really do not understand what CI is in the first place — which again points out insufficient training, not a flaw inherent in targeted input. The examination should be placed on trainers and training, and on the willingness of teachers to invest the time required to become competent. There's no sense in saying root canal is worthless if you're looking at my attempts at doing it at home instead of looking at the results obtained by someone with training. Even though I know how to brush and floss.
The fact that people do not seem to like is that teaching with TPRS/CI requires effort to master. It requires skills. Skills require practice. This is not something where a teacher can go to a workshop on Saturday and pop into the classroom on Monday and do TPRS/CI full-blown and fantastic.
It's interesting to me that in a day when "grit" is the catchphrase for our students, we are not calling teachers out when they malign the method for a lack of training and practice. If we control for the effect of teacher competence in TPRS/CI, the differences between targeting types will most likely disappear, just as controlling for poverty in the US eliminates differences in achievement.
« Engagement and the Left-Handed Language Learner
Substituting ingredients »
Wow, this list of 100 English fairy tales are great to use for Level 1 TPRS stories!
Too many voices
When do you do Step 1?
The case against 是 (shi4)
An Online Class or Workshop Booking
Copyright © Optimizing Immersion | Powered by WordPress


modern drop ceiling image of modern drop ceiling tiles modern drop ceiling lighting.
modern drop ceiling modern dropped ceiling modern dropped ceiling image of basement drop ceiling ideas home design software modern suspended ceiling ideas modern kitchen drop ceiling.
modern drop ceiling drop ceiling lighting ideas drop ceiling lighting as lights for living room modern suspended recessed basement suspended ceiling lighting ideas images of modern drop ceilings.
modern drop ceiling wood drop ceiling modern drop ceiling ideas medium size of wood drop ceiling modern wood modern wood drop ceiling.
modern drop ceiling drop down ceiling ideas modern suspended ceiling drop ceiling panels drop ceiling ideas images of modern drop ceilings.
modern drop ceiling modern drop ceiling modern drop ceiling tiles useful links modern suspended ceiling systems modern drop ceiling modern suspended ceiling lights.
modern drop ceiling modern drop ceiling ideas basement best drop ceiling ideas modern drop ceiling modern drop ceiling tile.
modern drop ceiling modern drop ceiling alternatives office modern kitchen drop ceiling.
modern drop ceiling get quotations a led ceiling lights living room modern flowers bloom after atmospheric dining hall images of modern drop ceilings.
modern drop ceiling decorative drop ceiling photo 4 of modern drop ceiling tiles decorative drop in ceiling tiles 4 decorative drop ceiling modern drop ceiling tile.
modern drop ceiling drop light new ceiling light modern drop ceiling lighting new led lighting for modern drop ceiling lighting.
modern drop ceiling modern ceiling tiles modern lines faux tin ceiling tile gallery modern drop ceiling tiles modern ceiling modern kitchen drop ceiling.
modern drop ceiling home depot dropped ceiling tiles modern drop ceiling tiles in waterproof quality homes decor 5 home modern suspended ceiling tiles.
modern drop ceiling dropped ceiling ideas drop ceiling ideas drop ceiling design interior design walls and suspended ceiling ideas dropped ceiling modern drop ceiling tile.
modern drop ceiling modern nded ceiling drop designs ideas modern drop ceiling basement.
modern drop ceiling modern dropped ceiling modern drop ceiling lighting modern dropped ceiling ideas drop ceiling modern design.
modern drop ceiling modern drop ceiling google search images of modern drop ceilings.
modern drop ceiling modern drop ceiling modern drop ceiling alternatives modern drop ceiling basement drop ceiling modern design.
modern drop ceiling modern drop ceiling ideas modern wood drop ceiling.
modern drop ceiling modern drop ceiling tiles vast acoustic perforated aluminum ceiling tiles acoustic perforated drop modern drop ceiling options.
modern drop ceiling image of modern wood ceiling planks faux drop ceiling modern design.
modern drop ceiling modern dropped ceiling drop ceiling tiles wood panels homes quality designs inside ideas 3 home design modern drop ceiling options.
modern drop ceiling suspended ceiling living room design with suspended ceiling modern suspended ceiling tiles.
modern drop ceiling ceiling lights led panels for false ceiling discount ceiling lights suspended ceiling light fixtures prices drop ceiling modern design.
modern drop ceiling living room false ceiling designs design trends premium images of modern drop ceilings.
modern drop ceiling elegant modern drop ceiling lighting calm dining room with modern drop ceiling combined recessed modern drop ceiling tile.
modern drop ceiling bathroom ceilings modern drop ceiling tile.
modern drop ceiling dropped ceiling led lights drop ceiling in bathroom ceiling lights led fascinating drop ceiling lights full dropped ceiling modern drop ceiling lighting.
modern drop ceiling modern dropped ceiling modern drop ceiling modern drop ceiling designs with modern suspended ceiling tiles modern drop ceiling options.
modern drop ceiling photo of a modern open plan office with suspended ceiling images of modern drop ceilings.
modern drop ceiling led flush mount kitchen lighting fresh ceiling light modern drop ceiling lighting new led lighting for modern drop ceiling options.
modern drop ceiling drop ceiling with irregular patches of squares and rectangles modern dropped ideas modern wood drop ceiling.
modern drop ceiling dropped ceiling design drop ceiling design dropped ideas bat lighting dropped ceiling pics dropped ceiling modern drop ceiling lighting.
modern drop ceiling modern drop ceiling tiles decent best acoustical ceiling images on modern suspended ceiling lights.
modern drop ceiling image of wood drop ceiling panels x ideas modern drop ceiling options.
modern drop ceiling modern drop ceiling modern drop ceiling panels modern suspended ceiling lights.
modern drop ceiling modern lighting ideas lighting ceiling design outstanding living room with modern drop ceiling combined rectangle interior modern drop ceiling tile.


text:
Набережный — поселок в Арбажском районе Кировской области.

География
Находится на левом берегу реки Пижма на расстоянии примерно 6 километров по прямой на юго-запад от районного центра поселка Арбаж.

История
Известен с 1939 года как льнозавод Арбажский. В 1950 году учтено дворов 22 и жителей 54, в 1989 году проживало 108 жителей . До января 2021 года входил в Арбажское городское поселение до его упразднения.

Население
Постоянное население составляло 100 человек (русские 86%) в 2002 году, 66 в 2010.

Примечания

Населённые пункты Арбажского района

redpajama_set_name: RedPajamaWikipedia


text:
\section{Supplemental Material}

The final lepton $p_T$ distributions for individual analysis channels
are shown in Figure~\ref{fig:metqt}.
The uncertainty from each systematic source as a percentage of the final
background estimate for each channel is provided in Table~\ref{tab:syst}.

\begin{figure}[!b]
\begin{center}
\mbox{
\includegraphics[width=1.62in]{EPAPS_fig1a_color.eps} \hfill
\includegraphics[width=1.62in]{EPAPS_fig1b_color.eps}
}
\mbox{
\includegraphics[width=1.62in]{EPAPS_fig1c_color.eps} \hfill
\includegraphics[width=1.62in]{EPAPS_fig1d_color.eps}
}
\mbox{
\includegraphics[width=1.62in]{EPAPS_fig1e_color.eps} \hfill
\includegraphics[width=1.62in]{EPAPS_fig1f_color.eps}
}
\caption{\label{fig:metqt}
Distributions of the (a) leading and (b) trailing electron $p_T$ in the $ee$ channel,
(c) electron and (d) muon $p_T$ in the $e\mu$ channel, and
(e) leading and (f) trailing muon $p_T$ in the $\mu\mu$ channel after final selection.
Data are compared to estimated signal, $\sigma(WW)=12$~pb, and background sum.
}
\end{center}
\end{figure}

\begin{table}
\caption{\label{tab:syst}
Uncertainty from each systematic source as a percentage of the final background
estimate for each channel.
}
\begin{tabular}{p{2.0in}rrr}
\hline\hline
Systematic Source 			& $ee$   & $e\mu$ & $\mu\mu$ \\
\hline
MC Statistics 				& 2.6\%  &  3.6\% & 12.8\% \\
MC Cross Section			& 2.6\%  &  2.2\% &  2.5\% \\
MC Corrections				& 2.0\%  &  6.1\% &  2.0\% \\
Electron ID				& 0.3\%  &  0.3\% &  ---   \\
Muon ID					& ---    &  0.4\% &  3.0\% \\
$\gamma\to e$ Mis-ID Rate		& 1.2\%  & 12.1\% &  ---   \\
$W+$jet Statistics			& 15.5\% &  7.5\% &  6.8\% \\
$W+$jet $e$ Efficiency \& Mis-ID Rate	& 3.4\%  &  2.9\% &  ---   \\
$W+$jet $\mu$ Efficiency \& Mis-ID Rate	& ---    &  1.4\% &  2.1\% \\
\hline
Total Background Uncertainty		& 16.4\% & 16.4\% & 15.3\% \\
\hline\hline
\end{tabular}
\end{table}

In Table~\ref{tab:syst}, the ``MC Statistics'' uncertainty is based on the number of MC events used
to estimate the number of background events after final selection.  The ``MC Cross
Section'' uncertainty is based on the PDF and scale uncertainties
associated with the theoretical cross sections used to scale the
MC-driven background estimations.  The ``MC Corrections'' uncertainty
accounts for data-driven reweighting of the MC that corrects for general
event characteristics such as the primary vertex $z$-position and
instantaneous luminosity distributions.
The ``$\gamma\to e$ Mis-ID Rate'' uncertainty is driven
by statistics in the data sample used to determine that misidentification rate.
Sources of systematic uncertainty for the $W+$jet background estimation
are separated into categories based upon their correlations
across analysis channels.
The ``$W+$jet Statistics'' systematic is based on the number of
events in data that pass a series of lepton identification requirements
in each analysis channel, independently.
The ``$W+$jet Lepton Efficiency \& Mis-ID Rate''
uncertainties are based on measuring the rate at which a lepton passes
loose and tight identification in $Z/\gamma^*\to\ell\ell$ candidate events
in data and the rate at which a jet is misidentified as a lepton with loose or tight
identification based on dijet data.

The statistical uncertainty due to the number of observed events in
each channel is the dominant source of final uncertainty.

\vfill

\end{document}

redpajama_set_name: RedPajamaArXiv


text:
New partnership with MedTech Actuator will drive Australia's future growth
by Turbo Dev | Oct 13, 2020 | News
Australia has long punched above its weight in scientific production. The most recent figures from the OECD ranked Australia 9th
globally in 2015. We must continue to build capacity to commercialise this scientific production to support a strong Australian economy – and recovery.
The Menzies Foundation and the MedTech ActuatorTM
have partnered to do just this. The MedTech Actuator Menzies Fellowship will create the jobs of the future while also recognising the potential for scientific research to drive Australia's future growth.
Two MedTech Actuator Menzies Fellowships – each valued at $70,000 – will be awarded to high-potential, career life sciences researchers, giving them training in entrepreneurship and leadership. As part of the 12-month program, recipients will be immersed in the MedTech Actuator's industry-led model of commercialisation. They'll receive mentoring, take part in a tailored accelerator program and receive a $20,000 stipend.
"Through our accelerator, the MedTech Actuator provides intensive mentorship, industry collaboration, and capital raising support to MedTech, HealthTech and BioTech startups allowing them to achieve in 15-months what can typically take three years or more," says MedTech Actuator CEO, Dr Buzz Palmer.
To move a medical technology innovation through commercialisation and onto the market requires a network of experts in many areas including regulatory and reimbursement strategy, intellectual property, clinical trials and product development. And this complexity is often where aspiring entrepreneurs come undone.
"The MedTech Actuator Menzies Fellowship will provide high-potential scientists with a grounding in the elements of commercialisation, but most importantly it will provide access to our dense network of partners – a network that spans hospitals, product development firms, multinational MedTech corporations and investors," says Dr Buzz Palmer.
The Menzies Foundation believes philanthropy can play a unique role in sparking discovery and innovation in Australia. We are passionate about investing in our country's future science leaders and giving them the runway to ensure that their research has an impact in the world. We look forward to sharing their entrepreneurial journey," Menzies Foundation CEO Liz Gillies said.
The aim of the program is to support new leaders, drive strong and lasting entrepreneurship training, ecosystem connections and exchange. "We hope that our MedTech Actuator Menzies Fellows will take what they have learnt and the contacts they have made back to their organisation. It is by developing these connections that we'll build the technologies, economy and jobs of tomorrow."
The MedTech Actuator Menzies Fellowship is supported by the Menzies Foundation as part of their Entrepreneurship in Science mission in partnership with the MedTech Actuator.


This poison may be either driven out or drawn out.
almost invariably ruin the digestive organs.
pores of the tender skin on the bottom of the feet.
The Drafts absorb this poison.
their regular practice, and recommending them.


text:
info@futureangler.org
Home ContactAdd RBFF Plugins to Your Website DONATE
Learn About Fishing through Hands-on Experiences

"Getting Families Fishing"

Fish Art Contest Casting For Entries
Date:February 17, 2021
Author:Future Angler
Comments:0
The deadline is fast approaching for the 2021 Art of Conservation™ Fish Art Contest, supported by Title Sponsor Bass Pro Shops. This free contest is open to youth in grades K-12 from across the globe. The deadline to enter is March 31st, start your entry today!
Students from around the world can win top honors, international recognition and prizes while learning about fish, fishing and aquatic conservation. The Art of Conservation® inspires young people to get involved in the great outdoors and ignites a passion to become lifelong activists for the natural world.
Misty Mitchell, Director of Conservation Programs at the Johnny Morris Foundation said, "The Fish Art Contest allows youth the opportunity to connect with nature, no matter where they live. From the inner city to rural America, the Art of Conservation has the power to transform lives. Johnny Morris's Wonders of Wildlife National Museum and Aquarium is proud to support to a program committed to shaping the future of the outdoors."
To enter, artists create an original illustration of their chosen fish from the Official Fish List. Students in 4th grade and above should also submit a one-page creative writing piece based on the behavior, habitat or conservation needs of their species. Artwork and Essays will both be judged and awarded.
Julia Luger, Education and Communication Director of Wildlife Forever said, "The Fish Art Contest is a great opportunity for young people to connect with the great outdoors. Teaching youth the Art of Conservation is the perfect way to inspire the next generation of stewards."
Winners will be awarded in four grade categories, K-3, 4-6, 7-9, and 10-12. Entries must be sent by Wednesday, March 31st, 2021. Judging will occur in April and winners will be announced in May. See our 2020 winners and learn more at FishArt.org.
About the Artwork: Jessie X, 1st place winner, 2020 Pennsylvania Fish Art Contest, 7th-9th Grade Division.
About the Fish Art Contest: The award-winning Wildlife Forever Fish Art™ Contest, with support from Title Sponsor Bass Pro Shops, the USDA Forest Service and Guy Harvey Ocean Foundation, brings children, art and aquatic conservation together! The annual contest reaches thousands of youth each year. New distance learning resources allow students to participate from home or classroom and complement a wide array of educational programming. To enter, young artists create an original illustration of any fish from the Official Fish list and written words detailing its habitat, and efforts to conserve it. Entries are categorized in four grade levels: K-3, 4-6, 7-9, and 10-12. Educators nationwide utilize Fish On!, the full-color Fish Art Lesson Plan, integrating the disciplines of science and art. Entries are due postmarked or emailed by March 31st each year. www.fishart.org.
About Wildlife Forever: Our mission is to conserve America's wildlife heritage through conservation education, preservation of habitat and management of fish and wildlife. Wildlife Forever is a 501c3 non-profit dedicated to investing resources on the ground. Recent audits reveal that 94% of every dollar supports our award-winning conservation programs. Please, Join Today and learn more about the Fish Art Contest®, Clean Drain Dry Initiative™ and Prairie City USA® at www.WildlifeForever.org.
About Bass Pro Shops®: Bass Pro Shops is North America's premier outdoor and conservation company. Founded in 1972 when avid young angler Johnny Morris began selling tackle out of his father's liquor store in Springfield, Missouri, today the company provides customers with unmatched offerings spanning premier destination retail, outdoor equipment manufacturing, world-class resort destinations and more. In 2017 Bass Pro Shops acquired Cabela's to create a "best-of-the-best" experience with superior products, dynamic locations and outstanding customer service. Bass Pro Shops also operates White River Marine Group, offering an unsurpassed collection of industry-leading boat brands, and Big Cedar Lodge, America's Premier Wilderness Resort. Under the visionary conservation leadership of Johnny Morris, Bass Pro Shops is a national leader in protecting habitat and introducing families to the outdoors and has been named by Forbes as "one of America's Best Employers."
Tags:Art ContestBass Pro ShopsFish ArtWildlife ForeverYouth Events
Women to Tackle Inshore Saltwater Fishing March 26-27
Ladies, Let's Go Fishing University Announced for South Florida
Betty Bauman Wins Women Rocking the Web Award
New Anglers Become Winners at "Ladies, Let's Go Fishing!" Keys University Oct. 15-17, 2021
Youth Feel Like They Were The Head2Head Fishing PWS Champion
The Future Angler Foundation has recently received numerous messages, emails, and phone calls from individuals regarding an organization called the National Anglers Association. We'd like to clarify to all individuals who are looking for National Anglers Association contact information that we have no affiliation with this organization. We currently do not have contact information for the National Anglers Association. If we are able to find such information we will share that information via our website and on social media within this alert.
Patrick Neu, President
Future Angler Foundation
Forestville, Wisconsin
Tournament Programs
Outreach and Education
Supporting Partner Programs
Into the Outdoors
Add RBFF Plugins to Your Website
© 2020 Future Angler Foundation. All rights reserved.