text:
I wish I could come up with a title
…that would be as wonderful as the article I have to share with you today. I feel honored to "know" the woman who expressed everything I thought when I heard about this gaffe that President Obama made. Honestly, right now I couldn't have begun to do it the justice Johanna has. I feel blessed to count her among my "friends" – someone I've never met but know in a heartbeat I could reach out to. It's what makes us special parents even more special. An extended family as it were. I'd like to share this small piece by one of our own, our extended family. Johanna Mattern Allen.
Rethink your outlook on the disabled
By Johanna Mattern Allen
Posted: Mar. 23, 2009
Words cannot begin to express how disappointing it was to hear President Barack Obama's Special Olympics gaffe on "The Tonight Show with Jay Leno" on Thursday.
But it's not just Obama who needs to take a hard look at this. His remark on one of the most popular television shows in America is indicative of a culture that is all too comfortable disparaging individuals with a disability – I might add, individuals who never would disparage him or anyone.
Since he so decidedly put his foot in his mouth, here's my four-point plan for the president to make reparations for the cultural damage he caused last week:
* Pony up beaucoup bucks for the Special Olympics cause.
* Create a cabinet position for disability now. There currently isn't even a policy adviser for disability since Kareem Dale moved to an arts leadership position. With a disability population in the United States of 50 million-plus, and growing especially as our population ages, we need disability experts to work alongside our president.
* Create positions for self-advocates in the White House so the president and the world never forget about people who have to work harder than he ever can dream of working to achieve what they do.
* Urge every college, university and high school in America to teach disability history/cultural competency.
Obama isn't the only smart (read: well-rounded intellectual) person I know who knows jack about disability. And not all of us are as lucky as me to have my son, Jack (who has Down syndrome), for a teacher.
It's totally cool to not know, but do something about it instead of getting defensive, making excuses or ignoring it. Here's my simple, pain-free, four-point plan for the rest of us:
* Read some disability history. Read Paul K. Longmore.
* When interacting with an individual with a disability, presume competence. Always. Just because someone moves, communicates, sits, eats, breathes, walks, hears, sees, thinks or problem-solves differently, or doesn't do any of these things, he or she still experiences life, contributes to the world, has feelings and thrives and depends on relationships with others.
* Don't defend offensive language. Just because it comfortably rolls off one's tongue in mixed company or it's self-deprecating or we've always said it, that doesn't mean it's right. The next time you think "we're being too sensitive," think about how you sound clinging to an outdated term and defending it after the minority group being maligned has asked you to stop. If you need to be self-deprecating, use a thesaurus. Find the word or phrase you like and practice it before you need it – that's how habits get broken. Language influences culture, culture influences policy and, in my son's case, he can hear you (and so can I).
* Give us a break. No really. Take the time to be with a parent of a child with a disability or an individual with a disability. Encourage your children to have a play date with a child with a disability. Challenge the idea of why you might not have a friend with a disability. Reach out in friendship to those of us who are most marginalized. The great secret about disability is that each one of us is only a heartbeat away from it at all times.
The great tragedy of past generations is that there have been unspoken divides between the cultures of the disabled and those who are not. In the culture of disability, we're accustomed to cheering on individuals with great challenges to help them overcome great obstacles and odds.
We in the disability community know those of you who aren't disabled are able to learn more and know you are capable of using inclusive language and joining us in a 21st-century way of thinking.
Johanna Mattern Allen lives in Milwaukee. March 31 is the Special Olympics' "Spread the Word to End the Word Day," a national day of awareness calling for America to stop and reconsider the use of the "r" word: retard/ed. Go to www.r-word.org
No related posts.


text:
End Caps
Your review: These fit perfectly to the cut ends of our Raychem heating cables. Great product!
Call us 7 days a week
Item #: WBB49348
Ships From US, Add 3-4 days For Delivery Estimated lead times do not include weekends or holidays. Transit time is determined by shipping method and destination. Orders requiring credit information might take additional time.
Customers Who Viewed This Also Viewed
Documents are in PDF format
Product Specifications
Avg. Customer Rating: (1 customer reviews)

subset: commoncrawl


text:
According to optical fiber industry survey, at the beginning of 2011 ~ 2013, GPON fiber optic transceivers module such as SFP transceivers, CWDM SFP transceivers,DWDM SFP transceivers and so on and so forth, which increased by the $11, fell to about $7 for quotation at present, a drop of 35%, the relative EPON quotation to see bones, has fallen beyond compare, GPON product price decline striking. Industry to estimate, this year China purchasing will be very positive in GPON, and in the national policy guidance, improve the bandwidth will be built to replace EPON with GPON.
Due to smartphone, tablet, or even IPTV Internet TV set-top boxes, will provide more and more audio and video data. For telecom operators, cable operators was 10 ~ 50 MBPS speed broadband,which is a shortage, therefore, no amortization of depreciation pressure of telecom operators in the Asia-Pacific region, will also directly are Gbps level above the GPON, let the future GPON fiber optic network access.
In addition to fiber optic communication industry in mainland China can provide GPON chip. In the United States and Japan the shares are higher.
Indirect orders received the mainland system, industry evaluation, fiber optic network telecommunications equipment factory in March this year of GPON Huawei procurement, have gradually warming, is expected by the first quarter of cautious wait-and-see attitude, into the second quarter of optimism.
Due to the high speed railway, optical fiber is a national state policy, therefore is expected that China mobile (HK), telecom operators, will not reduce investment in the fiber optic communication.


text:
Kristen Bell is "extremely co-dependent".
The 35-year-old actress - who has daughters Lincoln, three, and Delta, 16 months, with husband Dax Shepard - has suffered from anxiety and depression from a young age and admits her "bubbly" personality stems from a desperation for people to like her.
She said: "I'm extremely co-dependent.
"I shatter a little bit when I think people don't like me. That's part of why I lead with kindness and I compensate by being very bubbly all the time because it really hurts my feelings when I know I'm not liked. And I know that's not very healthy and I fight it all the time."
The problems run through the 'Frozen' actress' family and she was advised early on by her mother, a nurse, that she could be susceptible to a "serotonin imbalance" and gave her tips on how to combat the feelings of depression.
Speaking on 'Off Camera with Sam Jones', she said: "[My mom's] a nurse and she had the wherewithal to recognise that in herself when she was feeling it and when I was 18 said, 'If you start to feel like you are twisting things around you, and you feel like there is no sunlight around you, and you are paralysed with fear, this is what it is and here's how you can help yourself."
While she feels there is still a stigma attached to mental illness, Kristen insists she isn't ashamed to ask for help if she needs it and still takes medication to keep her depression and anxiety at bay.
She said: "If you do decide to go on a prescription to help yourself, understand that the world wants to shame you for that, but in the medical community, you would never deny a diabetic his insulin. Ever.
"But for some reason, when someone needs a serotonin inhibitor, they're immediately crazy or something."


text:
03:48 PM - November 5, 2014 by MMChrisS
If you aren't careful, Anthony Davis can become a player with just about any team if the NBA 2K15 opposing GM AI has anything to say about it.
While the trade logic doesn't seem nearly as over-the-top bad as it was last year, there are still several superstar for superstar trades which don't make much sense in the real world. On that same token, some of the trades I have been offered as the GM of OKC have been downright silly.
In year one, I was offered James Harden and Dwight Howard for Russell Westbrook and Serge Ibaka.
While yes, that trade is far from a bad trade for anyone, you just normally don't see the faces of franchises being swapped like that -- especially not on a regular basis. Another potential area of concern is that I'm not sure how many teams have their original draft picks anymore, with a few teams having three and even four picks (the Celtics) in my second draft and a couple (Phoenix and the Lakers) having exactly 0 draft picks -- despite the fact both teams were atrocious in year two.
So what are you seeing? Is my case of excessive trading an anomaly or are the opposing GMs still a bit too quick to pull the trigger on trades in MyGM?


text:
Carragher alarmed at Suarez stance
Jamie Carragher believes it is "laughable" for his old Liverpool team-mate Luis Suarez to be complaining about the club's reluctance to sell him.
Suarez claims the club agreed to let him leave Anfield if Liverpool failed to qualify for this season's Champions League.
The club refute that, and while Carragher claims the Uruguayan is "too good" for both Liverpool and Arsenal, and better suited to the likes of Real Madrid, Barcelona and Bayern Munich, he expects greater loyalty from the striker.
Carragher said in his Daily Mail column: "It didn't surprise me that Brendan Rodgers has made him train on his own. I never thought Luis would down tools in the way Fernando Torres did when he wanted to leave Liverpool - his attitude over the final 18 months became progressively worse - but now Suarez has given me reason to think again.
"To see it come to this is very dispiriting. A lot of good people have stood by Luis since he joined the club. Rodgers and Kenny Dalglish could not have done more for him. In the darkest times, we wore T-shirts to show we were standing alongside him, even if it was ill-advised.
"So it is laughable to hear he is unhappy with Liverpool for not letting him join Arsenal."
Suarez was banned by the Football Association for racially abusing Manchester United defender Patrice Evra, and again towards the end of last season for biting Chelsea's Branislav Ivanovic.
Despite those offences, Liverpool still want him at the club, however Carragher claims he has seen at close quarters a change in Suarez's attitude to life at Liverpool.
He said: "I took part in a training session at Melwood before Steven Gerrard's testimonial last Saturday and I was alarmed by the Luis Suarez with whom I was sharing a pitch.
"From his first day at Liverpool, I had been used to seeing a warrior who loved his football and charged around with great enthusiasm.
"On this occasion, however, his attitude was completely different. He didn't want the ball and had no interest in getting involved. He stood on the periphery, with shoulders slumped."
While Carragher spent his entire career with Liverpool, before retiring in May, Michael Owen moved away to join Real Madrid in 2004.
And Owen, who also retired at the end of last season, can understand why Suarez, at the age of 26, would want to leave to join a club playing at the highest level in Europe.
Owen told the Daily Telegraph: "People have this stereotypical view of a footballer that he's just greedy, no loyalty, but you can bet your bottom dollar that if Suarez wasn't as good as he was then Liverpool wouldn't have stood by him [during his misdemeanours]. Of course, you have to bear in mind the things people have done for you along the way, and have some gratitude, but you have one shot.
"Luis has not come through the ranks. He doesn't have a huge affinity with Liverpool but the fans have a huge affinity with him and expect that to be reciprocated. But he wants to go and be at the best club he can.
"This is where I can't quite understand it. If it's Arsenal [that is] the only option for him, I can't really understand why he would want to leave given the way the Liverpool fans feel about him and the team is virtually built about him. If Real Madrid came in for him, I could certainly understand that."


text:
Another dangerous error is the doctrine that denies the Deity of Christ, claiming that he had no existence before his advent to this world .
This theory is received with favor by a large class who profess to believe the Bible, yet it directly contradicts the plainest statements of our saviour concerning his relationship with the Father, his divine character, and his pre-existence. it cannot be entertained without the most unwarranted wresting of the scriptures. It not only lowers man's conceptions of the work of redemption, but undermines faith in the Bible as a revelation from God.
While this renders it the more dangerous, it makes it also harder to meet. If men reject the testimony of the inspired scriptures concerning the Diety of Christ , it is in vain to argue the point with them, for no argument, however conclusive, could convince them.
The natural man receiveth not the things of the spirit of God: for they are foolishness unto him: neither can he know them, because they are spiritually discerned.(Corinthians 2:14).
None who hold this error can have a true conception of the character or the mission of Christ, or of the great plan of God for man's redemption. Still, another subtle and mischievous error is the fast-spreading belief that Satan has no existence as a personal being, that the name is used in scripture merely to represent man's evil thoughts and desires.
The teaching so widely echoed from popular pulpits, that the second advent of Christ is his coming to each individual at Death, is a device to divert the minds of men from his personal coming in the clouds of Heaven.
For years satan has thus been saying, Behold, he is in the secret chambers" (matthew 24:23-26). and many souls have been lost by accepting this deception.
While God has given ample evidence for Faith, he will never remove all excuse for unbelief.
Check my Blog tomorrow for topic eight.


text:
NEX-7 New User Initial Thoughts
I've been trying to pist this as a review, but the system keeps telling me that there is a link broken - so here goes:
I may update this as I have only had the camera a few days. Bought to replace a Canon 550D, which was my second camera to a 5D Mk 2. I wanted something smaller, lighter and more adaptable to street photography. It certainly fulfills these and image quality is very good - probably beating my 550D. I am also impressed by the autofocus - which seems fast and reliable, ability to handle highlights/shadows, and high ISO performance.
I got it with the 18-55 kit lens and bought a 55-210. Both seem at least OK (first impressions are that the 18-55 is the sharper). It is also very interesting that Sigma is making some prime lenses for NEX.
Camera ergonomics are good and the screen and EVF are good (the EVF is not as good as an optical viewfinder - IMHO).
The camera loses marks for three reasons:
Menu system. This is not good. There is a LONG settings menu, which hides things like turning off the steady shot (when you use a tripod - the lenses do not have switches), and bleep (discreet photography). You can add functions to custom buttons , but not these, so you have no choice but to root round in the menu.
Manual. Sony - come on please - you must have written many of these now. There is a paper starter manual, but the main manual is on PDF. Not good when you are out and about. The manual is also poorly index and explained. EG the explanation for steady shot is where it is on the menu and that you can turn it on and off. Cool. This takes one page and gives no advice - eg should you use it when the camera is on a tripod? This manual is unacceptable on any camera, let alone one costing £1,000.
RAW converter. VERY slow (on an i5 computer with 8GB ram and W7), clunky and does not seem to do conversions properly. Fortunately Adobe ACR handles the files well, but I may struggle getting a profile for the 55-210mm. The Sony software needs sorting.
Overall it is a very good camera, let down by the software - firmware and hard and soft manual. If Sony can sort these out - and these are not the rocket science parts - I would score the camera much more highly (I would give it 3.5 if I could).
Best regards
Vince C
|Post (hide subjects)
|Posted by
|Jul 3, 2012
|Jul 3, 2012
|Jul 3, 2012
|Jul 3, 2012
|Jul 3, 2012
|Jul 4, 2012
|Jul 4, 2012


text:
Using a 10,000-vehicle database, Fleet Systems Pty Ltd says that of the popular six-cylinder models, the 3.0-litre V6 Magna and Toyota Camry were more fuel efficient than the larger Holden Commodore and Ford Falcon.
According to the company's chief executive, David Stockley, the figures showed a significant improvement in running costs of all of the six-cylinder vehicles, minimising the difference between them and similar four-cylinder models (referring to both the Magna and Camry fours).
"For example, there is only a $510 per year cost difference between the Magna six cylinder and four cylinder, and a $150 per year difference between the Camry's six- and four-cylinder vehicles," he said.
These differences would be more than made up by the better pricing of the six-cylinder versions when it came to sell them on the second-hand market.
The fleet figures show that the fuel consumption of the latest VT Commodore is in line with the previous VS model; the higher maintenance costs of the Fleet Systems figures was more a reflection of the earlier model having covered a larger distance.
The 4.0-litre Falcon EL had the highest fuel consumption and maintenance costs in the six-cylinder categories, but again, the maintenance cost reflected the greater distance covered by the model, which was replaced late last year by the AU Falcon.
The AU is expected to have lower fuel consumption.
All models in the database had automatic transmission and air-conditioning fitted.
Statistics from Dec 1998 Fleet Systems 100,000 vehicle database
Falcon EL 4.0 litre: average odometer reading 48,000km, fuel consumption 12.4litres/100km, maintenance $22/1000km.
Falcon AU 4.0 litre: average odo 5600km, fuel consumption 12.2litres/100km, maintenance n.a.
Commodore VS 3.8 litre: average odo 52,900km, fuel consumption 12.2litres/100km, maintenance $19/1000km.
Commodore VT 3.8 litre: average odo 37,100km, fuel consumption 12.2litres/100km, maintenance $17/1000km.
Magna VF 3.0 litre: average odo 35,100, fuel consumption 11.4litres/100km. maintenance $15/1000km.
Camry 3.0 litre: average odo 42,600km, fuel consumption 11.9litres/100km, maintenance $20/1000km.
Ford Mondeo 2.0 litre: average odo 35,800km, fuel consumption 11.1litres/100km, maintenance $16/1000km.
Holden Vetra 2.0 litre: average odo 27,200km, fuel consumption 10.1litres/100km, maintenance $21/1000km.
Magna TF 2.4 litre: average odo 43,900km, fuel consumption 10.8litres/100km, maintenance $17/1000km.
Camry 2.2 litre: average odo 34,900, fuel consumption 11.2litres/100km, maintenance $19/1000km.
FOOTNOTE: Mitsubishi has introduced a new Magna since these statistics were compiled. It has a 3.5 litre engine standard above the base model, and the four-cylinder version has been dropped.


text:
Episode 53: The One Five Years after They Met
Barry talks about how getting an editorial letter triggers pretty much every neurosis he's got. (Morgan chortles.) Barry accidentally reveals the true extent of his ego. (Morgan chortles again.) Morgan talks scary movies. (Barry cowers.) Morgan reveals foreign parenting tips. (Barry freaks out.)


text: [In Regards to the Final Fantasy XIII-2 Hype Page.] Yeah, sorry about that whole final fantasy flame war thing. I was just in one of my moods and seeing the forum post was just the tip of the ice burg. In my ramblings I had no intention of "trolling" it just sorta came out that way due to my short, short fuse. I'm generally an agreeable person its just usually 9 times out of 10 when I do post it's when I'm in a bad mood and see something on the forum I don't like and let loose the dogs of war so to speak. But I don't try to, it's just hard to supress that feeling of rage when something you once loved so dear turning into a big pile of crap in front of your eyes. I mean, pretty crap, but crap none the less. My brain cannot comprehend certain things like why it's still so accepted even though it's clearly obviously garbage. Then my fingers go autopilot and my brain shuts off and all I'm after is blood at that point. So to whomever got in that cross fire I do Apologize for it. Shit was said That I didn't even mean to say I was just trying to get a rise from people. I really should be above this sort of behavior, One of the moderators (not naming names) spoke to me on irc to calm me down. I guess I was in deeper possible hot water then I realized. That's what really made me step back at the situation and see what I was doing. Just making myself look stupid and ignorant, and really I don't think anyone wants that for themselves if they have any kind of self respect. Actually I didn't even realize it was a big deal until said moderator stepped up and told me, I was blissfully unaware it was what you call a "flame war". I hear the term but don't really take thought into it's meaning. Thank Him he did cause honestly, I don't even realize I'm actually offending people even if I'm trying some times I don't realize that it might actually work. I wish I had a program on my computer that would slap me for going too far. Oh yeah that's what my girlfriend is for. ha ha.


text:
|Home | About | Journals | Submit | Contact Us | Français
Conceived and designed the experiments: DB PV MC FB. Performed the experiments: IB PF JA AF GB. Analyzed the data: PF JA AF GB MC FB. Contributed reagents/materials/analysis tools: CT FB PV ECJ CC GH JJ. Wrote the paper: MC FB.
Inflammatory breast cancer (IBC) is an aggressive form of BC poorly defined at the molecular level. We compared the molecular portraits of 63 IBC and 134 non-IBC (nIBC) clinical samples.
Genomic imbalances of 49 IBCs and 124 nIBCs were determined using high-resolution array-comparative genomic hybridization, and mRNA expression profiles of 197 samples using whole-genome microarrays. Genomic profiles of IBCs were as heterogeneous as those of nIBCs, and globally relatively close. However, IBCs showed more frequent "complex" patterns and a higher percentage of genes with CNAs per sample. The number of altered regions was similar in both types, although some regions were altered more frequently and/or with higher amplitude in IBCs. Many genes were similarly altered in both types; however, more genes displayed recurrent amplifications in IBCs. The percentage of genes whose mRNA expression correlated with CNAs was similar in both types for the gained genes, but ~7-fold lower in IBCs for the lost genes. Integrated analysis identified 24 potential candidate IBC-specific genes. Their combined expression accurately distinguished IBCs and nIBCS in an independent validation set, and retained an independent prognostic value in a series of 1,781 nIBCs, reinforcing the hypothesis for a link with IBC aggressiveness. Consistent with the hyperproliferative and invasive phenotype of IBC these genes are notably involved in protein translation, cell cycle, RNA processing and transcription, metabolism, and cell migration.
Our results suggest a higher genomic instability of IBC. We established the first repertory of DNA copy number alterations in this tumor, and provided a list of genes that may contribute to its aggressiveness and represent novel therapeutic targets.
Inflammatory breast cancer (IBC) [1] is one of the most lethal forms of breast cancer because of its high metastatic potential [2]. IBC is classified T4d in the TNM-UICC classification. Diagnosis is based on clinical signs: edema, erythema, pain, breast widening, and induration [2]. Most cases are associated with a ductal type and a high histological grade [3]. The presence of tumor emboli in dermal lymphatics, although not mandatory for the diagnosis, is a pathological hallmark of 50–75% of IBCs, Emboli are non-adherent cell clusters that rapidly spread by continuous passive dissemination [4], thus favoring both distant metastasis and local recurrence. Despite progresses due to the multi-modality treatment [2], [5], the prognosis remains poor with a 3-year survival of ~50% [6]. New molecular therapeutic targets need to be identified to improve treatment and increase patient survival.
Molecular mechanisms underlying IBC are poorly known (for review, see [7], [8]). IBCs are more frequently estrogen receptor (ER) and progesterone receptor (PR) negative, ERBB2 and EGFR positive. They frequently present P53 alterations and WISP3 loss-of-expression [9]–[15]. They show high angiogenic and angioinvasive capacities and express high levels of angiogenic factors [16]. They frequently overexpress RHOC [17]–[20], E-Cadherin [18], [21], [22], and NF-κB pathway-related proteins [23], [24]. Recently, a role for eIF4G1 has been suggested in the formation of tumor emboli, pointing to the importance of translation control in IBC [25].
High-throughput molecular approaches have led to important insights in the molecular heterogeneity of non-IBC (nIBC), notably by identifying biologically and clinically relevant subtypes (luminal A and B, basal, ERBB2-overexpressing, normal-like) [26]. More recently, IBCs have been studied at the mRNA level using DNA microarrays [27]–[33]. The results showed the presence of the five molecular subtypes in IBCs, and the definition of IBC versus nIBC gene expression signatures. But the studied series remain relatively small, with 35 IBC samples in the largest one [27], [28]. DNA copy number alterations (CNAs) may account for phenotypic and expression differences between IBCs and nIBCs. However, in contrast to nIBCs [34]–[39], genomic imbalances have not yet been analyzed in IBC by using recent techniques such as array-comparative genomic hybridization (aCGH) or SNP-arrays. The rare genomic studies performed to date used low resolution methods [40]–[42]. Similarly, whole-genome integrated studies (associating analysis of DNA CNAs and mRNA expression levels) have been done in nIBC [34], [39], [43], [44], but never in IBC. Such approaches provide opportunities to better elucidate IBC at the DNA and RNA levels.
Here, we have studied and compared DNA CNAs and mRNA expression deregulation on a whole-genome scale in a large series of IBCs and nIBCs. To our knowledge, this is both the first high-throughput aCGH analysis of IBC and the first whole-genome integrated analysis comparing IBC vs nIBC. This is also the largest series of IBC profiled using high-throughput molecular analyses.
Pre-treatment tumor tissues were collected from 197 patients with invasive adenocarcinomas, who underwent surgical biopsies or initial surgery at the Institut Paoli-Calmettes (IPC, Marseille, France) between 1987 and 2007. Each patient gave written informed consent and the study was approved by the IPC "Comité d'Orientation Stratégique". Tumor samples were macrodissected and frozen in liquid nitrogen within 30 minutes of removal. Before RNA extraction, tumor sections were reviewed by two pathologists (ECJ and JJ) and contained more than 60% of tumor cells. The 197 samples comprised 63 IBCs and 134 nIBCs. IBC tumors were selected upon clinical criteria as T4d tumor. The control group (nIBCs) represented a mixture of early (121 samples, including 68 with pathological axillary lymph node involvement).and locally-advanced stages (13 samples), as found in previous studies [27]–[29], [32], [33]. Using only locally-advanced cases as controls would introduce a bias toward the discovery of molecular differences related to the difference of growth kinetics between IBCs (sudden onset and rapid growth) and nIBCs (long-standing evolution with slower growth). Immunohistochemistry (IHC) status was available on most samples for ER (positivity cut-off: 10%), ERBB2 (0–3+ score, DAKO HercepTest, with >1+ defined as positive), and P53 (positivity cut-off: 1%). Patients were treated according to standard guidelines. The main histoclinical characteristics are listed in Table 1. As expected, IBCs were associated with more unfavorable prognostic features and poorer survival than nIBCs.
DNA and RNA were extracted from frozen samples by using guanidium isothiocynanate and cesium chloride gradient. DNA and RNA quality was respectively controlled on polyacrylamide gel electrophoresis and on Agilent Bioanalyzer (Agilent Technologies, Massy, France).
From the 197 samples analyzed, DNA was available for 173, including 49 IBCs and 124 nIBCs. Genomic imbalances of the DNA samples were determined by using high-resolution 244K CGH microarrays (Hu-244A, Agilent Technologies, Massy, France). A pool of 13 normal male DNA was used as reference. Scanning was done with Agilent Autofocus Dynamic Scanner (G2565BA, Agilent Technologies). Data analysis was done and visualized with CGH Analytics 3.4 software (Agilent Technologies). Extraction of data (log2 ratio) was done from CGH analytics, while normalized and filtered log2 ratio were obtained from "Feature extraction" software (Agilent Technologies). Data generated by probes mapped to X and Y chromosomes were eliminated. The final dataset contained 225,388 unique probes covering 22,509 genes and intergenic regions according to the hg17/NCBI human genome mapping database (build 35). Data were analyzed using circular binary segmentation (CBS) as implemented in the DNA copy R/Bioconductor package [45] with default parameters to translate intensity measurements in regions of equal copy number, each region being defined by at least five consecutive probes. Thus, each probe was assigned a segment value referred to as its "smoothed" value.
We used two different threshold values (log2 ratio >|0.5|, and |1|) to distinguish low level from high level CNAs [43]. DNA copy number patterns were classified as "simplex" (very few CNAs), "complex sawtooth" (many low-level CNAs), or "complex firestorm" (low-level CNAs and recurrent amplifications) [46], [47]. Unsupervised analysis was done with the Cluster program [48] using log2 ratio data, complete linkage and Pearson correlation as similarity metrics. Results were displayed using TreeView [48]. To determine the robustness of the resulting tumor clusters, we used R package Pvclust [49] with multiscale bootstrap resampling using 1,000 iterations. Approximately Unbiased (AU) p-values ≥90% were used to define the significant clusters. To identify altered regions, we used the GISTIC algorithm [50], which computes for each segment through the genome a score based on the frequency of CNA combined with its amplitude, with bootstrapping to calculate the significance level (p<0.05). To identify genes with different CNA frequencies between IBCs and nIBCs, the frequency of CNAs was computed for each probe locus as the proportion of samples showing an aberration therein. Alteration frequencies were compared using the Fisher's exact test and false discovery rate (FDR) was applied to correct the multiple testing hypothesis (p<0.05) [51].
Gene expression data of the 197 BCs and 4 normal breast (NB) samples, which represented 1 pool of 4 samples from 4 women, and 3 commercial pools of respectively 1, 2 and 4 normal breast RNA (Clontech, Palo Alto, CA), were quantified using whole-genome DNA microarrays (HG-U133 Plus 2.0, Affymetrix). Scanning was done with Affymetrix GeneArray scanner and quantification with Affymetrix GCOS software. Data were analyzed by the Robust Multichip Average method in R using Bioconductor and associated packages [52]. Robust Multichip Average did background adjustment, quantile normalization, and summarization of 11 oligonucleotides per gene. Before analysis, a first filtering step removed from the dataset the genes with low and poorly measured expression as defined by an expression value inferior to 100 units in all samples. All data was then log2-transformed for display and analysis.
Before clustering analysis, a second filtering retained the 12,813 probe sets with the most variable expression across all samples. Clustering was done with the Cluster program [48] using Pearson correlation as similarity metrics and centroid linkage clustering. Results were displayed using TreeView program [48]. The molecular subtypes of breast cancer were determined using the single sample predictor (SSP) classifier based on the list of 306 intrinsic genes as described [53]. The sample was attributed the subtype corresponding to the most correlated centroid. To develop a predictive model based on the expression of the 24 genes identified by integrated analysis (see below), we applied a logistic regression analysis using the glm function in R statistical package. Once defined, the model was applied to expression data to assign the "IBC-like" or the "nIBC-like" class: first for testing its robustness by leave-one-out cross-validation [54] in the learning set and by validation in an independent set of 24 IBC and nIBC samples, then for estimating its prognostic value in public nIBC datasets.
Genomic and expression data are MIAME-compliant (http://www.mged.org/Workgroups/MIAME/miame.html) and the raw data have been deposited in the MIAME-compliant GEO database (GSE23720).
Before integrated analysis, Affymetrix expression probe sets were matched with Agilent aCGH probes using the hg17/NCBI database. When multiple Affymetrix probe sets mapped to the same gene, the probe sets with an extension « _at », next « s_at », and followed by all other extensions were preferentially kept. When several probe sets with the best extension were available, the one with the highest median value was retained.
We analyzed the degree of correlation between RNA expression and CNAs for the genes and tumors (173 tumors) profiled with both platforms. For each gene with a CNA in at least two IBCs or nIBCs, a Student t-test compared the expression levels in samples with and without the corresponding CNA (FDR-corrected p<0.05). Comparative integrated analysis of IBCs and nIBCs was only applied to the genes with CNA frequencies significantly different between IBCs and nIBCs (628 genes). Genes had to satisfy three other criteria: i) frequencies of combined alterations (gain associated with overexpression vs other combinations, and conversely, loss associated with underexpression vs other combinations) different (Fisher exact test) between IBCs and nIBCs, ii) correlation (Student t-test) between CNA and expression in the 173 samples, and iii) expression different (Student t-test) between IBCs and nIBCs. In the first above-quoted step, overexpression and underexpression for a given gene were assigned using a threshold of |1| corresponding to twice the expression level found in the normal breast pool. These steps are summarized in Figure S1.
To test the prognostic performance of our 24-gene signature in nIBCs, we analyzed 6 public data sets collected from five publications [55]–[59], and from the UNC Microarray Database (Table S1). When different publications included the same patients redundancy was eliminated, resulting in 1,781 different patient samples available for analysis. Before analysis, we mapped hybridization probes for the 24 genes across the two oligonucleotide-based platforms used across the series. When multiple probes were mapped to the same GeneID (EntrezGene identification number), the one with the highest variance in a particular dataset was selected to represent the GeneID. Analysis of each data set (using available normalized data) was done separately to guarantee a larger number of genes common with our signature.
Correlations between sample groups and histoclinical factors were calculated with the Fisher's exact test for qualitative variables with discrete categories, and the Mann-Whitney test for continuous variables. Follow-up was measured from the date of diagnosis to the date of last news for patients without any event. Disease-free survival (DFS) was calculated from the date of diagnosis until the date of first relapse whatever its location (local, regional or distant) or date of death (when the relapse data was not available) using the Kaplan-Meier method. Survival was compared between groups with the log-rank test. Univariate and multivariate analyses were done using Cox regression analysis. The p-values were based on the Wald test, and patients with one or more missing data were excluded. All statistical tests were two-sided at the 5% level of significance. Analyses were done using the survival package (version 2.30), in the R software (version 2.9.1).
We first describe the results on the whole set of tumors before addressing the specific question of IBC. High-resolution aCGH was performed on 173 samples, including 49 IBCs and 124 nIBCs. Figure 1A (left) shows the frequency of low level CNAs in all samples. As previously reported [37], [44], the three most frequently gained regions were on 1q, 8q and 17q chromosomal arms, whereas the regions frequently lost were on 8p, 11q and 16q. The median percentage of probe sets displaying a CNA in a sample was 3.5%, with a great variability between samples (range, 0.03–44%). As expected, this percentage was higher in grade 3 tumors (2.1%) than in grade 1 tumors (0.5%; p=0.005; Mann-Whitney test).
To display the similarity of samples with respect to their genome profiles, unsupervised hierarchical clustering was applied to aCGH data of all 173 samples and all 225,388 oligonucleotide probes (excluding X and Y probes) (Figure 1B). Pvclust algorithm identified three robust tumor clusters, which correlated with the molecular subtypes of samples, and other histoclinical features (Table S2). No correlation existed with the IBC/nIBC status, suggesting similar whole-genome genomic profiles in IBCs and nIBCs, and a similar heterogeneity. Using a previous classification of genome profiles [39], [47], 55 cases (32%) were "simplex", 39 (23%) "complex sawtooth", and 79 (45%) "complex firestorm". This status correlated with the grade of tumors. Only 13% of grade 3 tumors were "simplex", whereas 87% were "complex". Conversely, 61% of grade 1 tumors were "simplex" and 38% "complex".
The 173 samples and 24 additional samples (197 including 63 IBCs and 134 nIBCs) were profiled using whole-genome mRNA expression microarrays. Figure S2 shows the hierarchical clustering of samples based on the expression of 12,813 probe sets. Samples were sorted into three major clusters, which strongly correlated with histoclinical features (grade, IHC data) and molecular subtypes, but not with the IBC status. As with aCGH analysis, differences due to molecular subtype greatly overcame differences due to clinical type. IBCs were scattered across the three clusters, intermixed with nIBCs, suggesting that, at the RNA level and on a whole-genome scale, they are as heterogeneous as nIBCs.
Our first objective was to establish the first repertoire of genome CNAs in IBC. Table S3 shows the regions with frequent low and/or high level CNAs in the 49 IBCs, defined using the GISTIC algorithm with a significance threshold of p<0.05. Sixty-five regions were gained/amplified. The top ten regions (in term of median GISTIC score) resided on 17q, 6p, 1q, 8q, 11q, 19q, and 8p chromosomal arms. Many of these regions contain genes involved in breast cancer, such as ERBB2, MYC, CCND1 and FGFR1. Thirty-four regions were gained in at least 15% of IBCs. The most frequently gained region (>40%) was 8q12.1–24.3, and contained 461 genes. A total of 216 regions were lost/deleted, with the top ten (in term of median GISTIC score) distributed in 4q, 3q, 1q, 9q, 15q and 2q. Only 4 regions were lost in at least 15% of IBCs.
At the gene level, 321 genes were amplified in at least 10% of IBCs (Table S4). As expected, all were located in the 65 regions gained/amplified reported above. They included validated or potential therapeutic targets such as ANGPT1, ERBB2, FGFR1, GRB7, MYC, PAK1, PNMT, PROSC, SQLE, and STK3. Other targets such as ADAM9 (6% of cases), EGFR, FNTA, and IKBKB (4%), AKT3, AREG, BRAF, EREG, FYN, and RET (2%) were less frequently amplified, but sometimes at a very high level (log2 ratio >|1|). The 15 genes that were the most frequently amplified are located in 17q12 and 8q24 (Table 2). Additionally, rare high-level amplifications targeted other potential therapeutic targets such as DCK, FGF3, FGF10, FLI1, IL6, MFHAS1, ROS1 and SRC. Conversely, 21 genes were deleted in at least 10% of IBCs (Table S4; Table 2), and all were included in the 216 lost/deleted regions.
We compared the genomic profiles of 173 samples (49 IBCs and 124 nIBCs). Globally, as shown in Figure 1A, IBCs looked like nIBCs, since both phenotypes showed similar altered regions with similar frequencies of alterations for most of them. However, some differences were visually apparent, such as the more frequent gain of 1q, 8q and 17q regions in IBCs, or the more frequent loss of 4p, 8p, 11q, and 16q regions in nIBCs. The median percentage of probe sets displaying a CNA for a sample was higher in IBCs (3.7%, range 0.01–14%) than in nIBCs (1.9%, range 0.01–26%; p=6.1.E–253, Fisher's exact test), even if a great variability between samples existed for both types. IBCs showed a higher proportion of "complex sawtooth" and "complex firestorm" profiles than nIBCs, which conversely showed a higher proportion of "simplex" profiles (p=0.0009, Fisher's exact test) (Figure S3).
The comparative analysis of regions with frequent low and/or high level CNA in IBCs vs nIBCs showed very similar results in term of number of altered regions (71 and 77 gained respectively, and 210 lost in both types) (Table S5). Many altered regions were similar. Some of them, such as 4q13.2 and 11q11, were similarly altered (similar GISTIC scores) whereas others, such as 1q21.2–q41, 8p11.21–p12, 8q11.1–q24.3, 11q13.33–q14.1, 17q11.1–q21.2, and 20q13.2–q13.33 were differentially altered (different GISTIC scores) either because of a higher frequency and/or higher amplitude of alteration. Other regions were specifically altered, such as 5p15.33 in IBCs or 3q26.1 in nIBCs.
The number of genes amplified or deleted was higher in IBCs than in nIBCs (Table S4). Only 26 genes were amplified in at least 10% of nIBCs, vs 321 in IBCs. As expected, these genes are located within amplicons classically described in breast cancer on 8p, 11q and 17q. Sixteen genes (vs 21 in IBCs) were deleted in at least 10% of nIBCs. Supervised analysis identified 9,148 probes, representing 628 unique sequences/genes, with different CNA frequency in IBCs and nIBCs (Tables S6A–B). A total of 514 genes displayed gains and/or amplifications differentially associated with IBCs vs nIBCs, including 484 genes gained/amplified more frequently in IBCs, and only 30 gained more frequently in nIBCs (Table S6). Compared to nIBCs, IBCs were associated with the gain of 382 genes scattered through 13 chromosomal arms (1p, 4q, 5p, 6p, 6q, 7p, 8q, 10q, 12q, 14q, 17q, 19p, and 20q). Figure S4 shows an example of genomic profiles for the 6q21 region. The frequently gained 8q21.2–24.3 region included more than 230 gained genes, including MTDH (8q22.1), RRM2B, AZIN1, FZD6 (8q22.3), ANGPT1, EIF3E (8q23.1), DCC1, MTBP (8q24.12) and ATAD2, SQLE (8q24.13). By contrast, only 30 genes, scattered through two chromosomal arms (3q and 16p), were more specifically gained in nIBCs, including MYH11, C16orf63 (16p13.11), ERCC4 (16p13.12), and A2BP1 (16p13.3). Thus, specific gains were more frequently observed in IBCs than in nIBCs. The mean frequency of tumors displaying a gain for one gene of these respective lists was 31% for IBCs (maximum 53% for genes located in the 8q22.3–24.11 region), and only 9% for nIBCs (maximum 10% for genes located in the 16p13.11 region). A total of 189 genes were more frequently amplified in IBCs than in nIBCs. Most of them were located within five chromosomal arms (1q, 8p, 8q, 17q, and 18p). Several genes such as RAD21 (8q24.11), MTBP (8q24.12), MYC, PVT1 (8q24.21), ERBB2 (17q12), and CASC3 (17q21.1) are known cancer-related genes. Ninety-nine of these 189 genes were amplified in at least 10% of IBCs. The mean frequency of IBCs with amplification for one of these genes was 14% (maximum 29% for genes such as ERBB2 located in the 17q12 region). In contrast, no gene was more frequently amplified in nIBCs than in IBCs.
A total of 114 genes displayed genomic losses and/or deletions differentially associated with IBCs vs nIBCs, including 68 genes lost/deleted more frequently in IBCs, and 46 more frequently in nIBCs (Table S7). Fifty-nine genes were more specifically lost in IBCs, affecting 7 chromosomal arms (4p, 5q, 6q, 12p, 15q, 19q, and 22q). Examples include EMB (5q11.1), RPS5, UBE2M (19q13.43), SLIT2 (4p15.31) and EZR (6q25.3). Conversely, 46 genes were more specifically lost in nIBCs, affecting two chromosomal arms (8p, and 16q). Examples include PKD1L2 (16q23.2), FOXC2 and FOXF1 (16q24.1). Loss and amplification of BC028701, FKSG2 and KCNU1 (8p12), were correlated with nIBCs and IBCs, respectively. The mean frequency of tumors displaying a loss for one gene of these respective lists was 11% for IBCs (maximum 24% for genes located in the 6q27 and 22q13.1 regions), and 20% for nIBCs (maximum 21% for genes located in the 16q24.1 region). In IBCs homozygous deletions (no copies assuming a modal diploid genome) targeted 9 genes located on five chromosomal arms (3q, 8p, 13q, 14q, and 18q). Four of them were deleted in at least 10% of IBCs. The mean frequency of IBCs with deletion for one of these genes was 10% (maximum 14% for genes located in the 8p23.1 region). No homozygous deletion was more frequently found in nIBCs.
Thus, among the 628 genes (514 gained/amplified and 114 lost/deleted) with CNAs differentially represented between IBCs and nIBCs, 552 (484 and 68; 88%) were associated with IBCs and 76 (30 and 46; 12%) with nIBCs.
We compared the degree of CNA-driven mRNA up and downregulation in 49 IBCs and 124 nIBCs profiled on both platforms by analyzing the 13,127 genes common to the two platforms and retained after filtering based on the expression levels.
In IBCs, from all genes with a CNA (gains/amplifications and losses/deletions) in at least two samples, 10.4% of gained genes exhibited mRNA expression correlated with the gain, and 1.5% of lost genes exhibited expression correlated with the loss (Table S8). In nIBCs, these respective features were 10.4% and 9.5%. For the gained genes, the percentages of correlations were similar in IBCs and nIBCS but for the lost genes, this percentage was 7-fold more important in nIBCs than in IBCs (p<1.E-12, Fisher's exact test; OR=0.14 (CI95% 0.09–0.20)).
To identify potential IBC-specific candidate genes, we compared in the 173 IBCs and nIBCs only the genes with deregulated mRNA expression in relation to CNA (Figure S1).
Out of the 628 genes identified by supervised analysis with CNA frequencies different between IBCs and nIBCs, 500 were present on the Affymetrix microarrays. They were represented by 748 probe sets on these microarrays, and 4,259 probes on the Agilent microarrays. From these 500 genes, we identified 24 genes whose expression was deregulated in relation to CNA with significant differences between IBCs and nIBCs (Table 3; Table S9; Figure 2). In all cases, the transcriptional deregulation was associated with IBCs only and corresponded to an overexpression related to a gain (21 genes) and/or amplification (13 genes). By definition, these 24 genes were also overexpressed in IBCs as compared to nIBCs. Twenty of these genes are located in 8q22–q24 and 17q21, including PAPBC1, RAD21, ATAD2, MTSS1, SQLE, ST3GAL1, C17orf37, ABCC3 and PTPN2, previously reported as cancer-related genes. In contrast, no candidate gene was both downregulated and loss-targeted in IBCs.
To validate the association of these 24 genes with the IBC/nIBC distinction, we analyzed the 24 remaining breast tumors (14 IBCs and 10 nIBCs) only profiled on our Affymetrix platform. None of them had been included in the 173 tumors from which the 24 gene-list had been derived. First, we defined an IBC/nIBC genomic classifier by applying logistic regression to the expression data of the 24 genes in the 173 samples (learning set). As expected, the rate of accurate resulting classification was high (86%). Leave-one-out cross-validation gave similar results with a rate of 82% (p=2.94E-11, Fisher's exact test; Figure 3A). Then, we applied the predictor to our validation set of 24 samples (Figure 3B): 75% of samples (10/14 IBCs and 8/10 nIBCs; p=0.03, Fisher's exact test) were correctly classified, suggesting the robustness of the classifier.
Because of the differences between IBC and nIBC regarding some histoclinical features such as grade, ER, ERBB2 and P53 status, we verified that our 24-gene signature was more associated to the IBC/nIBC distinction than to any of these features. This was done by comparing the p-values testing the correlations between the "IBC-like"/"nIBC-like" classes and each feature (Figure 3A–B). This was further confirmed by regression analysis on the whole set of samples testing and comparing the ability of each variable (24-gene signature, grade, ER, ERBB2 and P53 status) to discriminate IBC from nIBC (Table 4). Three features were significant in univariate analysis (24-gene signature, ERBB2 and P53 status), and two, including the 24-gene signature, remained significant in multivariate analysis. Altogether, these data indicated that our 24-gene signature is more linked to the distinction IBC/niBC than to other potential confounding variables.
Given the poor prognosis of IBCs, we hypothesized that the 24-gene predictor, if biologically relevant with respect to the IBC/nIBC distinction, might be prognostic in breast cancer. We thus tested its prognostic value in a series of 1,781 clinically annotated nIBCs. Using our 24-gene model (the number of genes common with each data set ranged from 17 to 24), we attributed to each sample an "IBC-like" or "nIBC-like" profile. All series were then pooled, resulting in 338 nIBCs with an "IBC-like" profile and 1,323 with a "nIBC-like" profile (Figure 3C).
We compared the DFS of these two nIBC classes for the 1,420 patients with available clinical outcome. With a median follow-up of 91 months after diagnosis (range, 1 to 220), the "nIBC-like" class had a 5-year DFS of 73%, better than the survival of the "IBC-like" class with a 5-year DFS of 61% (p=4.4.E-4, log-rank test). Survival curves are shown in Figure 3D.
We then performed univariate and multivariate DFS analyses (Table 5). In addition to the "IBC-like" or "nIBC-like" profile, we tested the variables most frequently annotated in the six data sets: patients' age, pathological tumor size, axillary lymph node status, and grade, and IHC ER and PR status (ERBB2 status not available). All features, except age and PR status, were significant in univariate analysis. The hazard ratio (HR) for relapse was 1.42 for "IBC-like" tumors compared to "nIBC-like" tumors ([95%CI 1.17–1.73], p=4.7.E-4). In multivariate analysis, these five variables, including the 24-gene model-based classification, retained significant prognostic value, suggesting that our IBC signature is an independent prognostic feature in nIBC, with a HR for relapse for "IBC-like" tumors vs "nIBC-like" tumors equal to 1.34 ([95%CI 1.06–1.70], p=0.015). This observation confirmed that the 24-gene signature contained some prognostic information, which might explain the worse prognosis of IBCs.
We applied high-throughput molecular analyses to a large series of IBCs and nIBCs. Because copy-number changes drive a considerable proportion of the transcriptional changes [34], we compared genome copy number and expression profiles to identify potential IBC-specific candidate genes. To our knowledge, this is the first high-resolution aCGH study of IBC, the first integrated genomic analysis for IBC vs nIBC comparison, and the largest series of IBCs profiled using high-resolution genomic analytic tools.
Because of the high number of CNAs observed in breast cancer, we used stringent log2 ratio threshold values to define the most specific genomic aberrations. We did not separate IBCs from nIBCs on the basis of whole genome profiles, which were globally very close, suggesting that IBCs are as heterogeneous as nIBCs at the genome level and that different obvious genome alterations are not what distinguish them. However, the number and frequency of CNAs were more important in IBCs than in nIBCs, as well as the proportion of "complex sawtooth" and "complex firestorm" profiles, clearly suggesting that the genomic differences between them are not due to a possible damping of the nIBC signal by higher contamination with normal tissue. Among the 628 genes with CNAs differentially represented between IBCs and nIBCs, 88% were associated with IBCs and only 12% with nIBCs. These results indicate a higher degree of genomic instability in IBCs, in agreement with their high grade, frequency of P53 mutations and their aggressiveness. Given this genomic complexity and heterogeneity of IBCs, and the low degree of differences observed with nIBCs globally, future studies should ideally compare IBCs and nIBCs within molecular subgroups defined by expression (molecular subtypes) and/or by CNA (simplex, complex sawtooth, complex firestorm) profiles as recently suggested [60]. In this context, and given the scarcity of IBC, international collaborations are underway for collecting enough IBC samples that should allow identifying genomic alterations perhaps more specific of IBC.
However, even if the number of profiled samples is relatively small, our series represents the largest tumor set reported in literature, and its study provides several interesting results. We identified 65 regions with gain/amplification and 216 with loss/deletion in IBCs. Few data are available in literature regarding the genomic imbalances in IBCs [40]–[42], and none has been done using high-resolution aCGH. A study of loss of heterozygosity at 71 microsatellite markers in 66 IBCs [41] reported deletions of various chromosomal regions including 3p14–p21.2, 6p, 8p22, 11q22–q23, 11q24–q25, 13q14, and especially 17q21, more frequently than in nIBCs. Our results show some overlap with these results (Table S3). We here identified four 17q21 genes, NSF, ARL17P1, ARL17 and KIAA1267 targeted by loss in 17% of IBCs (Table S3).
In both IBCs and nIBCs the number of regions with CNAs was very close and many altered regions were similar, suggesting that common genes are involved in both types. However, some common regions displayed higher frequency and/or higher amplitude of alteration in IBCs, and other regions were specifically altered in IBCs such as 5p15.33, previously associated with poor prognosis in breast [61] and bladder cancers [62].
At the gene level, the results were similar. Many genes were similarly altered in IBCs and nIBCs in terms of frequency and amplitude of aberrations. However, a larger number of genes displayed recurrent amplifications in IBCs than in nIBCs. Some genes, such as CCND1 and FGFR1, were amplified at the same frequency in the two types, whereas others (99 out of 321 recurrently amplified, 31%), such as ERBB2 and MYC, were more often amplified in IBCs. Of note, all high-level CNAs differentially represented between the two types were associated with IBCs, and none with nIBCs. Several of them code for validated or potential therapeutic targets, which could contribute to enlarge our therapeutic armament in IBC. The difference between IBCs and nIBCs was much less important regarding the genes recurrently deleted. The deletion of the RB1 tumor suppressor gene likely contributes to the genomic instability in IBC.
Whole-genome gene expression profiling did not distinguish IBCs and nIBCs more than did aCGH. To our knowledge, the information about the relationship between gene CNAs and mRNA expression in IBC is scarce in the literature. In the present study, we restricted the analysis of expression data to genes with DNA CNA. Having identified the genes with a CNA in at least two IBCs and in at least two nIBCs, we determined those whose mRNA expression correlated with the CNA. In both tumor types, ~10% of gained genes presented such correlation, in agreement with previous observations obtained with low resolution techniques and less samples in nIBC series [34], [63]. In contrast, this percentage for the genes with loss/deletion was smaller in IBCs (1.4%) than in nIBCs (9.5%), suggesting that epigenetic mechanisms might be more operational in IBCs than in nIBCs.
Integrated analysis of aCGH and expression data identified 24 genes as potential IBC-specific candidate oncogenes, whereas no IBC-specific gene inactivated by loss was found. This does not rule out the likely existence of IBC-specific tumor suppressor genes inactivated by other mechanisms, as well as the existence of a gene expression signatures identified by the sole comparison of whole-genome expression data of IBC vs nIBC. Importantly, the discriminative power of the predictive model - built from the expression levels of these 24 genes - was validated in an independent sample set. Furthermore, this model was an independent prognostic feature in a multicentric series of 1781 nIBCs, indirectly validating its association with IBC, known to be more aggressive than nIBC.
Whether these genes are causative or even predictive of the IBC phenotype in a biological sense or reflect aggressiveness or another associated phenomenon remain to be explored by further in-depth experimental analyses. Several encode proteins involved in the protein translation and transport: MRPL27, a component of mitochondrial ribosomes; VPS13B, involved in vesicle-mediated sorting and transport of proteins within the cell; ABCC3, an ATP-binding cassette transporter, and PABPC1 [64]. Increasing evidence points to a crucial role of translational regulation in cancer development and progression, notably in IBC [65]. PABPC1 is a poly(A)-binding protein (PABP) required for translation initiation. Its interaction with the translation initiation factor eIF4G is crucial for the translational stimulatory effect conferred by the poly(A) tail. In IBC, eIF4G1 reprograms the protein synthetic machinery for specifically increasing the translation of certain mRNAs, notably that encoding p120 catenin, resulting in an increased stabilization of E-cadherin, and that encoding VEGF [66, Silvera, 2009 #2199]]. E-cadherin stabilization maintains the structure of tumor emboli, allowing them to survive and to metastasize as entire structures. VEGF expression accounts for high levels of angiogenesis in IBC and resistance to hypoxia. Our result suggests that PABPC1 could also participate and potentiates this process, allowing IBC cells to adapt to the persistent hypoxia they experience as tumor emboli. Other genes are associated with cell cycle progression: RBM13/MAK16 [67], TAF2, ATAD2, UTP23, MTBP, and DSCC1. TAF2 is a general transcription factor particularly involved in the G2/M transition [68]. ATAD2, as target of E2F, ER and coactivator of MYC [69], links the 3 corresponding pathways, and likely contributes to the aggressiveness of disease through the enhancement of MYC-dependent transcription [70]. UTP23 [71] is a component of the small subunit processome, required for ribosome biogenesis and cell cycle progression at G1. MTBP regulates the E3 ubiquitin ligase activity of MDM2, a critical negative regulator of p53 function [72]. DSCC1 is a component of an alternative replication factor C complex that loads PCNA onto DNA during S phase. Genes of the signature encode proteins involved in RNA processing and transcription: the TAF2 and ATAD2 transcriptional regulators, ARID2, which facilitates ligand-dependent transcriptional activation by nuclear receptors [73], UTP23; INTS2 and INTS8, subunits of the integrator complex, which associates with RNA polymerase II and mediates 3-prime end processing of snRNAs (Baillat, D, Cell 2005). Other genes are associated with metabolism. ST3GAL1 encodes a glycosyltransferase that induces aberrant glycosylation of MUC1 in breast cancer [4], [74]. SQLE encodes a key enzyme of cholesterol biosynthesis, whose expression is associated with poor survival in nIBCs [75]. Two genes, RAD21 and RAD54B, are associated with DNA repair [76]. Finally, four genes (PTPN2, MTSS1, EPN3, and C17ORF37) are associated with cell migration and adhesion and/or poor prognosis of breast cancer. The PTPN2/TC-PTP phosphatase stimulates the ERK pathway [77], and its decreased expression markedly impairs IGF2 induced MCF7 migration [78]. The presence of MTSS1, which encodes for metastasis suppressor 1 [79] appears paradoxal, given its favorable prognostic impact in breast cancer [80]. However, it has been suggested that MTSS1 is unlikely to be a metastasis suppressor, but interacted with RAC, actin and actin-associated proteins to modulate lamellipodia formation [81]. Epsin 3 (EPN3) is involved in extracellular matrix-epithelial cells interactions [82]. C17orf37, whose expression correlates with grade and stage of nIBC [83], promotes invasion and migration of prostate cancer cells by enhancing secretion of uPA, MMP9 and VEGF through NF-kB pathway [84]. Altogether, these different processes (protein translation and transport, cell cycle, RNA processing and transcription, metabolism, cell migration) are consistent with the hyperproliferative and invasive phenotype of IBC. Some of them (protein processing, RNA translation, proliferation and lipid metabolism) have been previously reported as overrepresented among genes or pathways associated with IBC [29], [31].
In conclusion, we report the first description of genomic profiles of IBCs, on a large sample size and with a high-resolution aCGH platform, and the first integrated genomic analysis comparing IBC vs nIBC. This repertoire of whole-genome CNAs in IBCs may serve of basis for further investigations. We show the genomic complexity and heterogeneity of IBCs, which globally look like nIBCs. Many genes targeted by CNA - some of them specific of IBC - have not been previously reported in breast cancer. We have identified 24 IBC-specific potential oncogenes that could explain, at least partially, the IBC phenotype and its aggressiveness, and lead to the development of new therapeutic strategies. As such, they represent new candidates for further clinical and functional validation in IBC. One of them, PABPC1, is particularly interesting as it likely potentiates the role of an alteration recently discovered as essential in IBC pathogenesis. Our findings, as well as the comprehensive database of CNA and mRNA expression generated, constitute a novel step towards the goal of better understanding, and perhaps treating, IBC, even if other alterations of the tumor, as well as those of its microenvironment, at other molecular levels such as DNA mutations, epigenetic regulations, microRNAs [85], proteins and others [4], [25] still need to be identified.
Integrated comparative analysis of IBcs and nIBCs with the three successive steps numbered 1, 2 and 3).
Whole-genome expression profiling of IBCs and nIBCs. A) Hierarchical clustering of 197 samples and 12,813 probe sets with significant variation in mRNA expression level across the samples. Each row of the data matrix represents a gene and each column represents a sample. Expression levels are depicted according to the color scale shown at the bottom. Red and green indicate expression levels respectively above and below the median. The magnitude of deviation from the median is represented by the color saturation. The dendrogram of samples (above matrixes) represents overall similarities in gene expression profiles and is zoomed in B. Colored bars to the right indicate the locations of 6 gene clusters of interest (ECM means extra-cellular matrix). B) Dendrograms of samples. Top, three large groups of tumor samples (designated I to III) are evidenced and delimited by orange vertical lines. Below the dendrogram, are some histoclinical and molecular features of the samples: from top to bottom, clinical type (green for nIBC, and orange for IBC), SBR grade (white for grade I, grey for II, and black for III), IHC ER, ERBB2, and P53 status (white for negative, and black for positive), and intrinsic molecular subtypes (dark blue for luminal A, light blue for luminal B, red for basal, pink for ERBB2-overexpressing, and green for normal-like).
Proportion of genomic patterns in IBCs and nIBCs.
6q21 gains are more frequent in IBCs than nIBCs. Profiles of chromosome 6 show higher 6q21 gain frequency in IBC tumors than in nIBC (A). Regional genomic profiles were established with CGH analytics® software (Agilent Technologies), for IBC and nIBC cases (panels B and C) both within the genomic interval [105.9–114.7 Mb] of the long arm of the chromosome 6 (hg17 human genome mapping; build 35 from NCBI). Profiles are distinguishable by different colors corresponding to different cases. Several IBC cases showed 6q21 gain or regional or focused amplification (B), whereas only two nIBC cases displayed a regional amplification (C).
Description of the public nIBC data sets.
Histoclinical correlations of the three aCGH-clustered groups.
Regions with CNAs in IBCs.
Genes with high level CNAs in IBCs and/or nIBCs.
Regions with CNAs in nIBCs.
Genes with copy number gain/amplification frequencies significantly different between IBCs and nIBCs.
Genes with copy number loss/deletion frequencies significantly different between IBCs and nIBCs.
Correlation between DNA CNAs and RNA expression for the 13,127 genes present on both platforms, in IBCs, and in nIBCs.
List of 24 genes with gain or amplification correlated with overexpression showing significant frequency differences between IBCs and nIBCs.
Competing Interests: The authors have declared that no competing interests exist.
Funding: This work was supported by Inserm, Institut Paoli-Calmettes, and grants from Ligue Nationale Contre le Cancer (Label DB) and Institut National du Cancer (ACI2007, AO2008, AO translationnelle 2007 IBC07). IB has been supported by a fellowship from the Algerian Government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


text:
Sometimes it's not you – It's the app
When I installed the app, that was where the joysticks appeared by default. And yes, the screen is upside down. That was also the default. You can change the screen orientation and the joystick placement, but I didn't think to do so at first.
If you try to keep your left thumb on the yellow joystick, you inevitably grab the edge of the tablet with your hand. This means that your palm will be resting on top of the 3 standard Android menu buttons, which causes all sorts of bugs.
This design flaw slipped by me at first because I thought the app was simply buggy. It is buggy, but most of the problems came from where the controls were placed.
The reason I posted about this app was that I wanted to get you thinking about the apps you use. This was a very basic game and yet the developer still managed to muck up the interface. I'm hoping that this will inspire you to think about the apps you have trouble using. Perhaps it's not you; it could be a clunky interface design.


text:
|• Taylor Aronson is fifth in the league in plus-minus at +28.
• The Cyclones lead the league with 3.44 goals per game.
• Cincinnati hosts Kalamazoo on Wednesday and Wheeling on Friday and Saturday before traveling to Toledo on Sunday.
|• Mike Clemente is fourth with 1,100 saves.
• Evansville leads the league on the power play at 19.9 percent (53-for-267).
• The IceMen are at Fort Wayne on Friday and Kalamazoo on Saturday.
|• Steven Summerhays made 19 saves to record the win in his pro debut on Sunday.
• The Komets are the league's fifth-least penalized team with 13.19 penalty minutes per game.
• Fort Wayne hosts Evansville on Friday and Toledo on Saturday.
|• Jordan Binnington is third among rookie goaltenders with a 2.33 goals-against average.
• Kalamazoo has a league-leading eight wins when trailing after two periods.
• The K-Wings are at Cincinnati on Wednesday and Toledo on Friday before hosting Evansville on Saturday.
|• Hannu Toivonen has started 13 of the last 15 games.
• The Walleye are 2-1-1 in their last four home games.
• Toledo hosts Kalamazoo on Friday, visits Fort Wayne on Saturday and hosts Cincinnati on Sunday.


text:
News and Information
Vacation Policy 3433
2017-2018 Draft Calendar
A bill was passed in the most recent WV Legislative session which allows accrued instructional time to be used to make up snow days, late arrivals/early dismissals and to provide staff development. Although this legislation has been signed into law, County Boards of Education are still awaiting guidance from the West Virginia Department of Education on the proper way to incorporate the use of accrued instructional time into the school calendar. Therefore, PLEASE BE AWARE THAT THE 2017-18 CALENDAR POSTED ON THE MORGAN COUNTY SCHOOLS WEBSITE IS SUBJECT TO CHANGE ONCE THE GUIDANCE IS RECEIVED. A final version of the calendar will be published as soon as it is available.
Quick Links
Sprigeo Reporting


text:
Athens folk songsters The Warblers will celebrate the release of ''Mama's Waltz,'' the long-awaited follow-up to their self-titled 1992 debut CD.
They'll perform from 7-8:30 p.m. Thursday in Masters Hall at the Georgia Center for Continuing Education. Admission is $6; call (706) 542-9842 for more information.
Published in the Athens Banner-Herald on Tuesday, December 18, 2001.
© 2017. All Rights Reserved. | Contact Us


text:
Former Tennessee Rep. Harold Ford Jr. — a black Democrat who was chairman of the Democratic Leadership Council — is in the running to get a transportation or infrastructure post in the Trump Administration, according to a source.
The source said Ford is a good friend of Donald Trump Jr., Ivanka Trump and her husband Jared Kushner, and is itching to get back in government.
Ford served in Congress for five terms until 2007.
He's now working at Morgan Stanley.
"Harold is happy with what he's doing," a friend of Ford told The Post.
"But if the president-elect of the United States calls, you take the call and listen carefully. Anyone would," he added.
Ford, 46, is a frequent political commentator on MSNBC's "Morning Joe," a show watched by the president-elect.
Ford comes from a political family and his father, Harold Ford Sr., served 11 terms in Congress. Ford successfully ran in Tennessee for his father's congressional seat—upon his retirement—and easily won at the age of 26.
After losing a Senate race in his home state to Republican Bob Corker, who himself is said to be in the running for a Trump Administration position, Ford moved to New York City.
In 2010, Ford considered running against Sen. Kirsten Gillibrand, but ultimately decided not to.
Ford did not respond to a request for comment.


text:
The Millers' tale of style
FRESH from receiving critical praise for her return to the West End, Sienna Miller has launched a collection of clothes with older sister Savannah.
They were in Selfridges, central London, to present the latest range of their Twenty8Twelve label.
Miller ( 29) is in a production of Terence Rattigan's wartime play, 'Flare Path'. Miller was lambasted by critics when she played Cecilia in 'As You Like It' in 2005. But her portrayal of a former actress in a love triangle was described as "impeccable". She stars alongside Sheridan Smith, who won the award for Best Actress in a Musical at the Olivier Awards.
Irish Independent


text:
Progressive TV and radio star Thom Hartmann took time off from covering Van Jones and his "Rebuild the Dream" movement on Wednesday to briefly talk to this columnist about his relationship with the Vladimir Putin regime of Russia. The conversation quickly went sour when Hartmann objected to questions about how much he is being paid by Moscow. He grabbed my video camera, covering the lens briefly in the process, and stomped away, objecting to "gotcha" questions.
The entire spectacle was captured on camera and posted on YouTube.
Advertisement - story continues below
During another embarrassing incident, Van Jones refused to sign a copy of his Standing Together to Organize a Revolutionary Movement (STORM) manifesto when asked to autograph it. "No thank you," he sternly said before asking, "Who are you?," looking at my name badge, and walking away to the next adoring crowd of Van Jones groupies. Jones, a "former" communist, is the new face of the progressive movement that is backing the "Occupation Wall Street" protests and is trying to guarantee President Obama a second term. He was fired as Obama's "green jobs czar" when the truth about his extremist background and statements started to emerge in reports originally by blogger Trevor Loudon and then Glenn Beck on Fox News.
While Jones made remarks in his own conference speech about losing his White House job, he also mocked Beck, noting that he had lost his program on the Fox News Channel. Progressives demanded Beck's ouster after he began exposing billionaire hedge fund operator George Soros, a major funder of the liberal-left.
TRENDING: Military Hero Sings Anthem During Thanksgiving Game, Fans Notice NFL Player's Actions on the Sidelines
Soros, who operates an off-shore hedge fund with mysterious sources of cash, has just failed in his effort to have the European Court of Human Rights lift his conviction in an insider trading case in France.
Read More at Accuracy in Media By Cliff Kincaid, Accuracy in Media
Advertisement - story continues below


text:
Take a bow!
Going potty means you're a big chickie now!
A playful and helpful audiobook for all parents and children as they go through potty training!
©2014 Janee Trasler (P)2014 HarperCollinsPublishers
There are no reviews for this title yet.
Report Inappropriate Content
If you find this review inappropriate and think it should be removed from our site, let us know. This report will be reviewed by Audible and we will take appropriate action.


text:
Seiyou Kottou Yougashiten: Antique
Type TV
Airing Status Finished Airing
Duration 22 min. per episode
Air Date Jul 4, 2008
Season summer 2008
Producers Nippon Animation, SynergySP, Fuji TV, Right Stuf, Shirogumi, Sony Music Entertainment

A high school crush, a world-class pastry chef, a former middle-weight boxing champion... and a whole lot of cake!

 Ono has come a long way since the agonizing day in high school when he confessed his love to handsome Tachibana. Now, some 14 years later Ono, a world-class pastry chef and gay playboy has it all. No man can resist Ono's charms (or his cooking skills!) but he has just found a new position under a man named Tachibana. Can this be the only man who resisted his charms, and if so, will the man who once snubbed the "magically gay" Ono get his just deserts? And how in the heck did a former middleweight boxing champion wind up as Ono's cake boy?

 (Source: DMP)

Recent EpisodesAll Episodes
Anime Relations
No Relations Found.


text:
Seattle Sports Deluxe Deck Bag
Where to Buy
sponsored links
Help support this site by making your next gear purchase through one of the links above. Click a link, buy what you need, and the seller will contribute a portion of the purchase price to support Trailspace's independent gear reviews.
Retailers' Descriptions
Here's what other sites are saying:
With multiple tie-down points, bungees and side straps for added storage, the Seattle Sports Deluxe Deck Bag provides plenty of storage and easy access to your gear while you paddle.
Multiple tie-down points, bungees, and side straps provide room for more gear Dimensions: 6 x 13 x 14" Weight: 1lb, 12.8oz Capacity: 791 inches cubed RF-welded seams keep water out Reflective end and attachment point for a strobe mount provides more visiblility Features a clear window for finding items quickly High profile maximizes storage space Item Specifications keyword search Boat keyword search Canoe keyword search Kayak
- Mountains Plus
Seattle Sports Deluxe Kayak Deck Bag - The Deluxe Kayaking Deck bag by Seattle Sports features multiple tie-down points, bungees, and side straps for added storage. You will have plenty of room for added gear with this Deluxe Kayak Deck Bag. It features a clear window for easier gear location and a higher profile to maximize storage space. Fully RF-welded seams will keep water out, while a reflective end with lashing for strobe mount will help keep you visible
- Outdoorplay
Similar Products
Seattle Sports Deck Mate Deck Bag
Seattle Sports Mesh Deck Bag
Seattle Sports Parabolic Deck Bag
Seattle Sports Profile Deck Bag
Sea to Summit Solution Gear Access Deck Bag
$80 - $89
SealLine Baja Deck Bag
NRS Sea Kayak Mesh Deck Bag
$26 - $69
NRS Watershed Aleutian Deck Bag
Seattle Sports Deluxe Deck Bag
previously retailed for:
$19.93 - $63.90
$19.93 - $63.90
The Seattle Sports Deluxe Deck Bag is not available from the stores we monitor. It was last seen November 14, 2017 at REI.


text:
Rabbits Trailer (2002)
01 January 2002 Fantasy, Drama, Comedy 42 mins
A story of a group of humanoid rabbits and their depressive, daily life. The plot includes Jane ironing, Suzie sitting on a couch, Jack walking in and out of the apartment, and the occasional solo singing number by Suzie or Jane. At one point the rabbits also make contact with their "leader".
Watch the official Rabbits 2002 trailer in HD below or find more Rabbits videos on Vidimovie.


text:
Student Charter
Student Charter for International Programmes Students
The Student Charter states key mutual obligations between the University of London International Programme and it students. It has been developed in liaison with the Lead Colleges and through consultation with our students.
The Student Charter provides a guide to the services and behaviours which we believe that is reasonable for students and the International Programmes to expect from each other. The Student Charter is reviewed annually.
The 2015 Student Charter [PDF 5pgs,150KB] is available to download.
Please see the programme specification and detailed regulations for your programme. The University Regulations are available from University of London website [external link].


text:
Nothing new about insurers whining about cost of claims
Personal injury fraud is not the reason for rising premiums
We all know the ads. Emotive images showing what the insurers want you to believe is a culture of widespread personal-injury fraud. As a campaign, it is very effective, like the slick PR machine that the insurance industry employs to blame everybody but themselves for sharply-rising motor insurance premiums. It's time to debunk some of these myths.
Insurers are losing money, so they have to increase premiums
Like many of the excuses rolled out to explain hefty premium increases, they expect us to believe that the market is unprofitable.
Yet, the largest insurer, Aviva Ireland, has produced its best profits for five years, up 32pc to €91.2m, while continuing to put on the poor mouth.
Dorothea Dowling, ex-chair of the Injuries Board and of the Motor Insurance Advisory Board, has consistently asked the industry to provide evidence of the losses which they claim to have sustained on motor insurance. This would require disclosure and transparency. The silence has been deafening.
Court-awarded damages are increasing
This has no basis in fact and, indeed, it would appear that the opposite may be the case.
While reports of large court awards tend to attract media attention, the reality is a lot more mundane. The arrival of the Court of Appeal has had unexpected effect on awards in the High Court. In the last six months, four cases have been successfully appealed and resulted in plaintiff awards being effectively halved in each case.
While this may appear at first glance to be an indication that awards are too high, it actually has a deflationary effect in general terms. It is a signal that the new appellate court is being proactive in ensuring that compensation remains at a reasonable level.
The Setanta decision against MIBI is significant
Perhaps the most disingenuous claim of all. The Court of Appeal has determined that the Motor Insurance Bureau of Ireland (MIBI) is responsible for all 1,700 claims arising from the collapse of Setanta Insurance. While this has been appealed to the Supreme Court, it has not stopped hysterical claims of massive future premiums.
The Setanta claims will be paid for by either the MIBI (funded by all the insurers on a pro-rata basis), or the Insurance Compensation Fund (paid for by a 2pc State levy on all premiums). Either way, premiums go up, but only for this relatively small insolvency for which no one is responsible and will have a minimal effect on premiums. Depending on the Supreme Court ruling, if the MIBI lose, they just have to ask the Government to change the agreement to exclude future insolvencies and there is no reason for the State to refuse this request.
Whiplash damages are too high
Whiplash is a colloquial term often used in a pejorative sense to describe neck, shoulder and back injuries sustained in car accidents.
The condescending manner in which many commentators refer to what can often be a very debilitating injury is one of the more distasteful aspects of the debate.
Insurers whining about the cost of claims is cyclical. We were here in 2002 and their reward was the Injuries Board. Having got what they wanted, they are still not happy. Maybe this time, it is the consumer that should be casting a cynical eye on their exaggerated claims.
Stuart Gilhooly is senior VP of the Law Society and a litigation lawyer at HJ Ward & Co
Sunday Indo Business


text:
Canyons Of Night : Number 1 in series Paperback
by Jayne Castle
Part of the Rainshadow Island series
Charlotte Enright, owner of a small antiques shop called Looking Glass Antiques on Rainshadow Island, and Slade Attridge, the community's new chief of police, both have something in common: they possess strong paranormal talents.
They met several years ago when they were in their teens spending the summer on the island.
Slade saved Charlotte from a gang of drunken toughs, but then at the end of the summer Slade and Charlotte went their separate ways and started their adult lives. Now, fifteen years later, they have both been drawn back to Rainshadow Island.
They will discover the adult passion they have for each other and start to explore some of the mysteries of the forbidden section of the island known only as the Preserve.
- Format: Paperback
- Pages: 368 pages
- Publisher: Little, Brown Book Group
- Publication Date: 01/08/2011
- Category: Fantasy
Other Formats
- EPUB from £4.99
Free Home Delivery
on all orders
Pick up orders
from local bookshops
Showing 1 - 1 of 1 reviews.
Review by wyvernfriend
Honestly this doesn't read much like SF. Apart from the fluff bunny it really didn't come across as all that SFnal. Slade Attridge is the new chief of police on Rainshadow Island. His career investigating paranormal events for the government is over. His last case had his psychic abilities go funny on him and now he's unsure how much longer they're going to last. Charlotte Enright has taken over her aunt's antique shop, Looking Glass Antiques on the island. Trying to sort out the variety of antiques her aunt has left her with. They were attracted to each other when they were younger and now the attraction is probably stronger, after all, now they're adults.It's an interesting read, more about the couple than about the psionics and exploring more of the world they're living in. Interesting but there wes something that just didn't gel for me.
Also by Jayne Castle | View all
Also in the Rainshadow Island series | View all


text:
The Boating Public
Would you give your car keys to a guy that does not have a driver's license? You probably wouldn't. Next question, would you give your boat keys to a guy that does not have a boaters license. Again if you did not know any better your answer would be no. ". On average, approximately 700 people die in recreational boating accidents each year. It is the belief of boating safety experts that requiring recreational boaters to have boating safety instruction could save numerous lives each year" (uscgboating, 2008). I am arguing in favor of mandatory licensing or boating safety class certificate in order to operate a boat.
Since boating safety and education requirements have been a hot topic, mainly since the early seventies, there has never been a federally mandated requirement for mandatory boater education in order to operate a vessel. According to the 1988 Boating Safety Manual, there were no states that mandated operator licensing or boater education. Since then 33 states now require some form of boater education. The only states that require licensing are Alabama, Connecticut, and New Jersey (Moore, Sarver, 2000). Although these states have stepped up and made licensing a requirement they are age restricted. For example, Alabama's requirement is anyone born after 1954 is required to obtain a license to operate any type of boat in their waters. What they have done is affix a "V" designation to the normal driver's license to prove the person has passed the approved test. Basically a 55 year old man can go out and buy a boat and put it in the water and drive away with no form of boating course or anything. I bet if that same 55 year old man went in to get a car drivers license he would have to pass a written and practical test before he gets a license.
Why is it that we require people to get a license to drive a car and not a boat? They both are powered by an engine. There are even more aspects about driving a boat that make it...


text:
To quit smoking weed might be very easy…
…for some people, but a horribly difficult thing for others. Many people would like to quit smoking weed the sooner the better but simply just have no clue how to get started. Some try to quit anyway, without having any plan, or strategy or whatsoever – and fail grandiosely. But if you have smoked for a long time and want to free yourself from cannabis now, you need a working exit-strategy and I hope this article might give you some help and advice.
The debate about cannabis addiction is a very controversial one since many people are not able to differentiate between a real physical addiction and a mental dependency. But while you may face some physical withdrawal symptoms, quite similar to what cigarettes smokers experience, in most cases it´s rather the psychological aspects that most long-term users struggle with.
This is even truer when smoking joints each and every day has become a habit or routine to you and you have got used to live in a stoned mindset. So, even though you´re not really have a physical need for cannabis, you may quite likely have developed a mental need for being high. This one of the first things you need to get aware of: You're not addicted to weed, but you´re mentally addicted to be high.
From my personal experience these are the 3 most important steps if you want to quit smoking weed:
1. Set a Starting Date to Quit Smoking Weed
From I recommend setting your fix date for quit smoking weed, within maximum a month from now on. Setting a quit date allows you to get prepared to quit smoking weed both mentally and physically. Don´t make the mistake to keep it some kind of sketchy like next month, or when school starts again, or as soon as my bag is empty. This is how I´ve failed many times before I learned just how important the setting of an actual date is. I convinced myself more than once that this bag would be my very last, and blasted it just like there would be no tomorrow. But as soon as this bag has run out, I bought myself another last one, and another last, and so on… Know what I mean?
2. Prepare Your House to Quit Smoking Weed
Weed smokers often have loads of wonderful equipment. If you´re serious about quitting you want to get rid of it; either donate them to friends or simply throw them away. I know this is harsh but whatever you do, clean up your environment and eliminate all weed or pot related gadgets. Believe me, it´s much easier to resist the cravings that will arise with a certainty once there is nothing that reminds you about it right in front of you.
3. Prepare Yourself to Quit Smoking Weed
One of the most common reasons for folks to fail on quit smoking weed is because they´re not prepared for the withdrawal. But alone by setting yourself a fix quit date, you´ve gained a big advantage, since you know just when the withdrawal will begin, and you can prepare yourself.
Anxieties just like when you´ve run out of dope (but of course much stronger), a loss of appetite and problems to fall asleep are very common symptoms of withdrawal in the first week of quitting weed. Therefore, do some research on what to expect in order to be prepared mentally when you face these problems.
In most cases these symptoms disappear after a week or two and once you´ve made it this far, chances are you may succeed on quit smoking weed. Although this may not be the end of the road already, you´ve made the first and most important step towards a new and self-determined life. No one says it´ll be completely easy but with the right strategies everyone can quit smoking weed.
Looking for a proven strategy to quit smoking weed?
Then you may >>> check this out! <<<


text:
Search Results
Entries tagged with "
Advertising Standards"
November 29, 2016 4:25 PM
Waze Does Display Advertising Right tagged "Advertising Standards"
There was a time when I thought all display advertising was "meh." I have reasons (and deep) rationale to think this way. Few were around selling banner ads when they first came out, like I did. I was on the front... Read more
April 9, 201010:32 PM
The End Of Online Advertising Standards tagged "Advertising Standards"
Do online advertising standards work? Is there a real point to them? The online and interactive advertising industry will have you believe that this is the only way to get both advertisers and publishers on the same page (so to... Read more
February 9, 201010:49 AM
This Will Change Everything... Again tagged "Advertising Standards"
What do you make of articles that say, "this will change everything for Marketers?" The truth is, I'm just as guilty as the next Journalist, Blogger and Writer to publish thoughts like that. The truth is, those types of sayings... Read more


text:
Affiliate Program – join free and earn up to 30%
Affiliate programs are the best way to earn income online for many, many people. They do not require any capital expenses, do not require a website, and do not require the affiliate to have anything personally to sell.
All the development work, testing, e-commerce, etc., is done for you, already. With an affiliate program, you, the affiliate, simply advertise the company's products or services to your contacts or on your website, and the company providing the affiliate program does all the heavy lifting for you.
It's always a pleasure to help someone else on the road to success in the affiliate program.


text:
A One bedroom apartment set within this prestigious development in the heart of Stratford offering easy access into the Olympic Park, Westfield shopping centre and Stratford Station. The property benefits from spacious living throughout, offering 1 double bedrooms, master bathroom, open plan lounge with fitted integrated kitchen, luxury bathroom & good storage.
Building Description:
– Gym Facilities
– Assigned property manager to help with your day-to-day duties
– 24 hour concierge services for deliveries, visitors & emergencies
– Hyper-optic broadband
– Tesco right beside the building
– Minutes away from the Olympic village
– Huge Westfield shopping centre at a 10 minutes walk
– 10 minutes walking distance to Stratford Station
– 4 minutes walking distance to the nearest DLR station
– Other services available for an additional cost include parking, apartment cleaning, dry cleaning and furniture rental
Apartment Description:
– High end interior
– Fully furnished kitchen
– Kitchen comes with Bosch & Indesit integrated appliances with stylish crystal work surfaces
– Engineered wood flooring & carpets
– Bathroom contains tiles from the Laura Ashley Malvern collection and comes with Grohe bath/shower & basins
Stratford Halo is less than a 10-minute walk to the Westfield Stratford City, and with Stratford station less than a 10-minute walk away from home, you could be at work in Canary Wharf in just over 20 minutes or in The City in less than 30 minutes.
For more information or to book a viewing please:


text:
A walk anywhere!
I believe it is extremely important to know who you're dating before getting to close and personal, so for that reason walking or hiking is my ideal first date.
just get to know you.... go for a coffe or a long walk by the beach
TO be honest I'd prefer my first date should be on the beach to take long walks on the beach and get to know each other while having amazing conversations and having the chance to see the sun go down .
Taking a tour, swimming fun, cooking exhibition, sweating fun
I love to take a walk on the beach, and after then eat something good, this is a Good chance to speak to each other
My first date ideas, sit in front of him with smile, and be like a feathers in the wind or a floating leaf in a flowing rivers
I would love to go for a walk to one of my favorite local surf breaks in Goleta and have a picnic . A place that I visit often and means alot to me. Then go wine tasting after lunch or to a clippers game!
Dinner or the park I love swings guess that's my inner child...
Walking and hiking develops the mind mentally and physically while the hard work pays off spiritually
Let's take hike and find a secret place, a hidden waterfall, let's take a boat out and find an empty beach Or lets have a coffee and talk about life.
For a first date I would say going for a walk and getting to know each other.
Take a walk on the beach going to nice restaurant in listed to music jazz drink wine
Follow Us
About Us |
Contact Us / Career |
Service Agreement |
Privacy Policy |
Success Stories |
Luxury Lifestyle Blog |
Dating Advice & Safety Tips |
Press & Media |
MillionaireMatch does not conduct background checks on the members of this website.


text: Cress Williams played Lester in the episodes Undecideds and Election Day (Part I). He is best known for his recurring roles on the television programs Prison Break (starring Robert Knepper and William Fichtner), Hart of Dixie (starring Tim Matheson), State of Affairs, Friday Night Lights (starring Connie Britton), Grey's Anatomy, John Wells' ER. Veronica Mars, Providence, and Nash Bridges (starring Jeff Perry).


text:
Pizza Hut Basra Airport
Here's a link to this photo. Just copy and paste!
Here is direct link of this photo
Copy and paste the code below (HTML):
Copy and paste the code below (BBCode):
[URL=https://www.arrse.co.uk/community/gallery/photos/lets-hit-the-hut.1027/]Let's hit the hut[/URL] by [URL=https://www.arrse.co.uk/community/gallery/authors/pondlife.7695/]pondlife[/URL] on Army Rumour Service
Separate names with a comma.