text:
VATICAN CITY -- Pope Benedict XVI urged the world's bishops on Sunday to try to bring back Catholics who have left the church as he opened a three-week meeting to reinvigorate the church's evangelization mission.
Some 262 cardinals, bishops and priests from around the world are in Rome for the meeting, or synod, called to give impetus to the pope's efforts to re-evangelize parts of the world where Catholicism has fallen by the wayside.
The synod coincides with the 50th anniversary of the start of the Second Vatican Council, the 1962-65 meetings that modernized the church.
Benedict has long lamented that in Europe and the Americas, Catholics no longer practice their faith or pass it on to their children. That concern is reflected in the synod's working document that will form the basis of discussion over the next three weeks.
"There is a clear link between the crisis in faith and the crisis in marriage," the pope said.
The so-called "new evangelization" is a top priority for Benedict, who routinely laments how cultures in Europe and the West that were once profoundly Christian have become increasingly secular.
The church has been beset by competition from rival Protestant churches in Latin America, dissent from Catholics who oppose church teaching on homosexuality and desertions in the United States and Europe from Catholics fed up by years of sex abuse scandals.
"The church exists to evangelize," Benedict said in his homily, urging a new missionary spirit among the church's pastors to reawaken the faith among Catholics who have been baptized but no longer practice their faith.


text:
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subset: commoncrawl


text:
Located just a short drive from the entrance to the Walt Disney World park, this complex is close to Highway 192, where there is a diverse selection of bars, restaurants and shops. The complex has plethora of amenities which offer the rewards of a resort, coupled with the privacy and comfort of a private villa. Further attractions in the surrounding area include SeaWorld Orlando, Universal Studios, and the Florida Mall Shopping Centre. This beautiful, Caribbean-style community is ideal for families or groups visiting Orlando.
Guests arriving from Orlando Airport should take the South exit from the airport, signposted State Road 417, Kissimmee. Join SR 417 heading West (this is a toll road). Exit at junction 2 signposted Celebration, and at the end of the exit ramp, after passing through the toll booth, turn right onto Celebration Avenue and drive 0.5 km to the junction with US Highway 192. Here turn left onto Highway 192, and then right at the 2nd set of traffic lights onto Parkway Boulevard. Take the first left turn and the Welcome Centre where check-in will take place is on the right.
Room Facilities:
Direct dial telephone
Satellite cable TV
Tea Coffee
Washing machine
Iron and board
Central airconditioned
Central heating


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March 7 2011, 7:14 AM
RUBBER Review
Niels Matthijs
; Antwerp, Belgium
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Sometimes a film needs years before it reaches cult status, others attain it only based on a simple web trailer released (think Iguchi's Machine Girl). Rubber falls in the latter category. Ever since word got out on this film, pulp and genre fans have been eagerly awaiting the new Dupieux feature. Luckily it stands up to all expectations and delivers everything you might have wished for, and a little extra. Brace yourself to enter the life of a killing tire.
You heard that right ... this film is about a killer tire on the loose. The piece of rubber even has a name: Robert. Though it sounds pretty insane, every now and than a film like this does pop up. Not too long ago I watched Battle Heater Kotatsu (1990), a film about a killer table stove. These things just happen I guess. What's
crazy about Rubber is that Dupieux found the capital and people to turn this into something more than a juvenile, semi-amateurish feature length film. Now there's something that completely boggles my mind.
While the name Quentin Dupieux might not ring a bell, maybe his moniker Mr Oizo does. He's the electro/trash producer that made fame with Flat Eric (the yellow, head-bopping puppet) when it was first featured in a Levis commercial. He kept busy producing electro over the years, now he's also ventured into the world of film, Rubber being already his second feature (in 2007 he made Steak but that one completely fell off the radar). He did most of the work on Rubber by himself, granting him writing, music, cinematography and even editing credits. I guess he likes to keep control over the end result.
While the trailer and all other advertising material might have you believe this is a film about a killing tire, it's only half the truth. Robert is definitely the main attraction, but there's a whole absurdist addition of a live audience following the "film" from a distance (using binoculars to get a better view). A welcome diversion as 70 minutes of rolling tires would've been a bit much.
Visually Rubber turned out to be much better than I expected. There are a fair few typical genre shots here, showcasing Dupieux's love and knowledge of the genre, most notably a "behind-the-back" shot when the tire is looking into a hotel room. Little visual references like that add something valuable to the overall feel of the film. Rubber also convinces on a technical level. While Robert's traveling shots might not seem too impressive at first, it's actually quite an accomplishment to move the tire through these rough surroundings in such a believable way, even playing around with the lighting of the sun in the meantime. Good stuff.
The soundtrack too is pretty cool. Awesome sound effects whenever Robert is blowing stuff up using his telekinetic powers, some very nice tunes throughout and even some lovely misplaced music just for the fun of it. Dupieux's background in music is obvious, of which he makes good use. The entire soundtrack could've been a little tighter as a whole, but the result is more than satisfying.
The acting is the only really weak part of the film, not surprisingly the part where Dupieux had the least control. Stephen Spinella does a decent job, the speech at the start of the film is quite unforgettable indeed, but the rest of the cast does very little with what they are given. For what is basically a deadpan comedy, Dupieux should've picked a cast with a bit more talent for comedy. Guess he still needs some training when it comes to pushing his actors in the right direction.
Rubber's promotional material is somewhat misleading in the sense that it tries to sell the film as pure nonsensical pulpy fun, while the actual film is actually much more related to deadpan comedy heroics like Dai-Nipponjin. The humor comes from slow pacing and the somewhat serious approach towards its actual core story (a tire coming to life). Don't expect to see much beyond that in the way of pulp, because you might leave the film terribly disappointed.
The extra abstract layer is fun but could've been much better if Dupieux had used a more experienced cast. While the absurdity of the whole situation is pretty ingenious, the humor comes in stilted dialogues and badly acted scenes, doing absolutely no justice to the ideas behind it. A real shame, certainly considering the first monologue by Spinella and the absolute potential it held.
No doubt Dupieux's film is going to reach a substantial audience based on its trailer and some good hype alone. But if you go in expecting what the promo material seems to promise chances are you'll leave a disappointed man. Go with the flow of the film and you'll see that there is a lot more to Rubber than a tire going loose on a killer spree. Hidden behind some subpar acting work is an accomplished little abstract comedy that delivers more than it promises.
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text:
About the Book
An insight into my portfolio developed by myself, Joanna Cooper.
It consists of photography, graphics, imaging and some artwork I have previously created.
"All our dreams can come true - if we have the courage to persue them"
Features & Details
- Category Arts & Photography
Standard Landscape, 10×8 in, 25×20 cm
26 Pages
- Publish Date Jan 27, 2014
- Language English
- Tags Portfolio, art, university, photography, education, Joanna, Cooper, ICT
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Let us know when your receive great advice so we can thank our lawyers and point others with similar questions in the right direction.Got it!
My childrens father was sentences to 4yrs. in prison for robbery class 2 1 count he has severd 5mos. He was just approved for work release I was hoping you could tell me how long he may have to be in the work release program? We have made plans to move from this state to Georgia for a new start but I am going to need to know how to go about doing so? I need to know how I would go about doing so?What do I have to do legal wise to get this started?
The situations you are describing come under the authority of the Interstate Compact pertaining to the transfer of Supervision. Typically, this is initiated by the State where the person who is on release is currently supervised, who then contacts the State where you want to move. Unfortunately, the individual States can refuse to accept the transfer, and in our experience do when the person being supervised has never been a resident of that State. I suggest you need to tell the person who is being supervised to discuss this with his parole officer who can give him more specific information on how IL will want to proceed. Good luck.
This answer does not, nor is it intended to, create an attorney-client relationship; or, constitute either legal advice or attorney advertising. Rather, given the nature of this forum, it is offered solely for information purposes, as a starting point for you to use when speaking directly to a lawyer in your State. Do not assume that the legal conclusions I mention that pertain to NJ are applicable in your State. Since the facts of each case are different, it is critical for you to consult with qualified counsel with whom information can be shared and assessed under an attorney-client privilege, so that competent advice can be obtained on which you can make informed decisions. Contact an experienced criminal defense lawyer admitted to practice in your State before making any decisions about your case.
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text:
Return to Fiction
the Sister of Scarlet's Servant
Author's note: Completely random, but drafted. Was partially inspired by Blargel's fan-fic, Wanderings of a Misplaced Stranger. This is not the best fan-fiction in the world. However, I worked very hard on it, and even introduced a new character with a new ability that could plausibly fit into a Touhou storyline. The main character is loosely based off of the author. This is still in progress so check regularly on the BBS thread and enjoy! ^^;
Return to Fiction


text:
|27 Jul 2012
CREATIVE XFI SB 0460 do not run with WIN 7 64
Hi everybody.
I am looking for a solution that makes my X-FI -SB0460 run again.
Since I installed Win 7 it stopped to run.
No matter 32 or 64 bits.
I download drivers from Creative and didn't work.
My machine isn't a new one but is perfectly functional.
I apreciate any help.
Thanks a lot.
|My System Specs
|27 Jul 2012
Try this one from major geeks downloads |MG| Creative Sound Blaster X-Fi Driver Windows 7 2.18.0015 Download
|My System Specs
|28 Jul 2012
I did it, one of my attempts. Didn't work. I'll try again. Thanks.
Correction: I couldn't find it at Major Geeks, no driver available for my Sound Card
|My System Specs
|28 Jul 2012
If you have an Xfi, give it a try. Won't hurt with all others that you have tried that did not work. Set a system restore point first. If it messes something up you can do a System Restore using the restore point you set and the PC will be back to how it was before attempting to install the driver.
|My System Specs
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My speakers partly working.. I checked and ran Audigy test ..result below: Any Idea how to fix that please.. Thanks
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Can't seem to get my Zen mozaic to work althou it says it is compatible here Windows 7 Compatibility for Creative Zen Mozaic 4GB Black Media Player: Creative. Drivers, Updates, Downloads When I plug it in it keeps saying device failed to install. Upon looking in device manager it says "This...
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creative x-fi
here's a question hope to see if someone knows the answer. My gateway p fx gives audio trough hdmi fine by using the integrated in audio card. I'm looking forward to buy an X-FI card and was wondering if i connect the laptop trough hdmi will i run into some issues? will i hear audio even if i use...
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Welcome to the home of the Firelord's Legacy and Mirrorsmith Chronicles.
The worlds of Riom and of the Regium will thrill, enchant and entertain you. I hope you enjoy your visit and I am always open to suggestions to improve your experience here. You can sample some of my books, read a short story or two (exclusive to this website) and find out more about the worlds I create.
I also love to encourage other indie authors and you can find links to fresh new talents as well as reviews of established favourites.
The Firelord's Legacy is an epic, sword and sorcery series beginning with The Firelord's Crown.
Raethwin. The Red Witch. Perhaps the greatest sorceress of her era. She sought to save her world. Instead she doomed it. The Firelord's Crown, source of her untold power. It brought about the impending disaster but Tamilin, Master Healer and Seer, believes it may also be their salvation. There's just one problem. Raethwin's six companions fled when their homeland was destroyed and divided the Crown between them. Five made it to safety. The sixth vanished. Master Tamilin believes it may lie in the uncharted northlands and sends an expedition in search of it. The Firelord's Crown begins when Airen, a young healer and lone survivor of the venture, reaches the land called Dinith in search of the lost piece. Dinith, however, is a land in turmoil. Magic is forbidden, the glories of the past forgotten. Airen and his quest are not welcome but could Falath, the king's heir, be the ally that he desperately craves? Perhaps, but Falath, descendant of the Lost Ship, has his own secrets...
The Firelord's Crown is available NOW on Amazon.
Book 2 Firelord's Heir is also available on Amazon! Now a prince in exile, Falath faces an uncertain future in a land of strangers; some of them friends, others most definitely enemies. The epic tale begun in The Firelord's Crown continues. Falath and Airen negotiate the dangers of new realms as they journey to the mystical heart of Mandras. They have the missing piece of the Crown but others want it too and will stop at nothing to acquire it. Falath must learn to control and develop his gifts of maisterie but still mourns the loss of his kinsman, Rollo. Meanwhile, a strange, fair-haired man is discovered by rebel scouts and subsequently healed by Jessal, their charismatic leader. This Mirin has no memory of who or what he is but Jessal realises that he could hold the key to their salvation. Answers lie with Guild Master Tamilin and both parties travel to the Healers' Isle in search of them but Mandras is a very dangerous place indeed. In his own land Falath is a prince, here he is just a valuable commodity...
Firelord's Curse is the third enthralling volume of the Firelord's Legacy series which began with The Firelord's Crown.
Falath and Airen have escaped from the fortress of Dim-Locil only to fall into the clutches of the mysterious and dangerous Stone People. Rollo, meanwhile, has entered the Ghost Wood, home of the Eye of Chimerand, another piece of the Firelord's Crown. Shimarah, High Priestess of the Eye, is intrigued by this stranger but her father views him as a danger to his realm, a threat to be neutralised or eliminated. Further to the north, Darial, badly wounded during the attack at the Isle, desperately needs to reach the capital with his information about their chief adversary's identity. First, however, he must try to cross the Great River but it's guarded by the Blue Company, sworn enemies of the rebels. Will any of them reach Caer-Banc before its too late and Corinius' plans are accomplished? For their world's sake they must but the odds are stacked against them. And what of Maristan? Alone in the stronghold of the Usurper, pregnant with Rollo's child and completely at Malglint's mercy. Can she survive her bitter circumstances or is she fated to die as her father before her, burnt at the stake as a witch?


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This Festool 491952 W48 fine toothed saw blade is an extremely powerful TCT blade that features crosscut technology for accurate and neat cuts. The type of teeth on this Festool blade ensure ultimate safety as they prevent splinters occurring in wood and veneered wood panels. This blade is compatible with the ATF 55, TS 55 and AP 55 machines.


text:
VIA Promotes Driver Development
Chip manufacturer VIA has announced a strategic Open Source initiative for Linux driver development.
The company is looking to cooperate with the Linux community on programming accelerated drivers for 2D, 3D and video output on platforms with VIA CPUs. An official discussion forum will provide a network for developers and users. A new website will provide beta versions of software and a bug tracking system. The Taiwanese vendor is looking to release quarterly updates in line with kernel and major distribution release cycles.
VIA has announced the launch of its new website at http://linux.via.com.tw for later this month. The site will give programmers access to drivers, technical documentation and source code for the VIA CN700, CX700/M, CN896, and VIA VX800 chipsets. The executive director of the Linux Foundation Jim Zemlin said that he was "delighted" about VIA's announcement in the press release here stating that "This action will open up exciting new opportunities for accelerating open source innovation around VIA platforms and ultimately delivering a better user experience."
Issue 201/2017
Buy this issue as a PDF
Kolab Now Integrates Collabora Online
Kolab Now subscribers now have access to Collabora Online.
Endless OS, a Distribution Without Internet
A new version of Endless OS has been released.
GitHub Announces Open Source Friday Program
They encourage people to take time out and work on open source projects on Friday.
System76 Announces Their Own Distro, Pop!_OS
Pop!_OS is based on Ubuntu and uses the Gnome stack.
Linus Torvalds Talks About His Motivation
He never tires of working on the Linux kernel.
Debian 9 Stretches Its Wings
The new release of Debian has a very strong focus on security.
Trojan Turns Raspberry Pi into a Cryptocurrency Mining Device
Two trojans in the wild are targeting Linux machines.
Fedora 26 Beta Comes with New Features
The workstation comes with the latest Gnome desktop environment and support for many application build systems.
Raspberry Pi Foundation Merges with CoderDojo Foundation
The two foundations join forces to expand their efforts to reach young people.
Samba Security Hole Patched but Risk is Bigger
Millions of devices that use Samba Server are vulnerable to attacks.


text:
THE INCIDENCE of melanoma of the skin appears to be rapidly rising. The increased incidence may be partially attributable to increased detection resulting from screening. In 1992, approximately 32000 newly diagnosed cases and 6700 deaths are expected in the United States. Melanoma tends to occur in adults in the prime of their family and professional lives. Detection and surgical treatment of the early stages of this malignancy are usually curative. In contrast, diagnosis and treatment in late stages often have dismal results.
Traits associated with an increased risk of developing melanoma include multiple typical moles, atypical moles, freckling, history of severe sunburn, ease of burning, inability to tan, and light hair with blue eyes. Other factors include the presence of familial atypical mole and melanoma syndrome, disorders of DNA repair, and excessive sun exposure.
Efforts to increase public awareness of melanoma and its treatment without causing unnecessary fear present a
Goldsmith LA, Askin FB, Chang AE, Cohen C, Dutcher JP, Gilgor RS, Green S, Harris EL, Havas S, Robinson JK, Swanson NA, Tempero MA, Ackerman AB, Balch CM, Cascinelli N, Clark WH, Farmer ER, Guerry D, Houghton AN, Koh HK, Kopf AW, Kraemer KH, MacKie RM, Maize JC, Meyskens FL, Perednia DA, Piepkorn MW, Rigel DS, Rogers GS, Sagebiel RW, Sober AJ, Tucker MA, Wick MR, Zone JJ, Moshell AN, Blume E, Bray E, Elliott JM, Ferguson JH, Gregg MB, Hall WH, Henson DE, Katz SI, Lotze MT, Maize J, Safavi KH, Sober A. Diagnosis and Treatment of Early MelanomaNIH Consensus Development Panel on Early Melanoma. JAMA. 1992;268(10):1314-1319. doi:10.1001/jama.1992.03490100112037
© 2017


text:
The ObjectBrowser Bean uses Java reflection to generate a file-explorer-like GUI automatically for any Java object or set of objects. It incorporates a number of features aimed at making the UI useable, such as special treatment of array-like members as child objects, and representation of bean getter and setter methods as properties. Scripting can be used to add in additional methods to manipulate the target object.
SearchAssist is a simple but practical search engine application that uses a ternary search tree. It uses Java's dynamic loading feature to make the search engine highly customizable, and uses takes Mozilla bookmarks as input. A Swing UI allows users to enter search words and view the results.
Builder Xcessory PRO for Linux is the industry's most advanced graphical user interface builder for Motif. BX PRO provides a comprehensive visual reuse environment that speeds development of mission-critical applications. BX PRO integrates three best-of-breed tools to create an unparalleled application development suite.
Uxtobx processes user interfaces generated by the UIM/X Motif GUI Builder tool developed by Actional and converts them into an UIL file that ICS's Builder Xcessory (BX PRO) can read and manipulate. Conversion is accomplished by giving the tool a list of files on the command line, which are read in and parsed. The resulting BX PRO UIL file is written to stdout and can be saved using standard file redirection. Any C/C++ code saved in a .i file is moved into a file called usercode.c during conversion process. After conversion, the user starts BX PRO and reads in the generated UIL file.
Thinlet is a GUI toolkit, implemented as a single Java class, which parses the XML defeinition of widget hierarchy and properties of the GUI, handles user interaction, and calls business logic. It separates the graphic presentation (described in an XML file) and the application methods (written as Java code).
GraphPak is a programming library of 2D and 3D charting objects for the Qt toolkit. It provides software developers with a set of C++ objects to easily create charts or graphs that aid in the visual presentation of technical and business data. This release includes Bar, Line, Pie, Ring, Area, Hi-Lo, Box and Whisker, and Polar charts. It is based on the KD Chart product from Klarälvdalens Datakonsult AB.
Libsx -- the Simple X library -- is a lightweight wrapper library sitting on top of the Athena Widget set, as implemented in Xaw and variants (Xaw95 yields much nicer results than Xaw in comination with libsx). Libsx does not compete with GTK or Qt, but nevertheless allows you to write a lot of small (or bigger) programs painlessly, with no or little knowledge of X programming. With libsx, writing a simple program that opens a window, has a few buttons, and draws some graphics (lines, circles, bitmaps, etc.) is a matter of 5-10 lines of code, including the drawing code. Hello World is 2 lines. More complicated programs that respond to mouse clicks, have buttons, scrollbars, file browsers, a color selector, etc., can be written in 25-50 lines of code.
Bambookit GUI is a completely XML-scriptable user interface to build real-time interactive Web application front-ends. Applications occupy 100 Kb of device memory and run on any Java-enabled browser. Users can move windows, resize containers, scroll and sort tables, lists, trees, see real-time data display, use a layout manager, and more. All rendering and event handling is managed in the XML scripts.
2D / 3D Pie Chart provides both a client and server side solution for the incorporation of pie charts into Web pages. It provides versatile components so that Web authors and Java developers can easily build and publish dynamic and interactive Pie Charts. It includes many configurable features and functions for both applet and servlet development, and the documentation and examples are designed to give both a full understanding of every feature and enable you to implement both the applet and servlet quickly and easily.


text: Sorting type: Most visited Most visited Alphabetical | A category for articles relating to House Reed. Pages in category "House Reed" House Reed " Lord of Greywater Watch is a title held by the ruler of Greywater Watch in The Neck. The title... Lord of Greywater Watch House Reed (Histories & Lore) Subcategories Individuals affiliated with House Reed Retrieved from "http://gameofthrones.wikia.com/wiki/Category:House_Reed?oldid=64751" Categories: Noble houses Vassal houses Houses from the North


text:
I don't blog much and this post alone will probably suppress my readership list by ten percent, but I feel it is very important to point something out. Joshua Strebel, although a pretty good guy, is a big, effin loser! Always a bridesmaid and never a bride (although he secretly dreams about me in a tux) he has always been number 2, in more ways than one, for longer than I can remember. (get the number two comment, it's not just rankings but it's also double entendre about him being a big poop)
If you want to join our campaign in calling out Josh as a big effin loser, please add the following code to your website!
Effin Loser
Thanks, that will be all!
I love you Josh. Call me, we'll do beers.


text:
Transcriptome characterisation of
Plant genomics
The Chinese pine (
The increasing amount of transcriptome data on
A large collection of high-quality ESTs was obtained,
Conifers are widely distributed globally as the largest and most diverse group of gymnosperms
Despite the economic and ecological importance of the genus
In addition to the discovery of new genes and investigations of gene expression, thousands of simple sequence repeats (SSRs), single nucleotide polymorphisms (SNPs) and insertions and deletions (Indels) have been detected in transcriptome data
Moreover, the increasing availability of transcriptome data represents an excellent resource for comparative genomic analysis. Although there has been much work on the chloroplast DNA sequences (cpDNA) and mitochondria DNA sequences (mtDNA), based on phylogenetic analysis of
In the current study, we used the Roche 454 GS-FLX Titanium pyrosequencing platform to obtain a comprehensive transcriptome of
Transcriptome sequencing and
Prior to sequencing, the cDNA samples obtained from multiple tissues and individuals were normalised to increase the sequencing efficiency of rare transcripts. Subsequently, 911,302 raw reads with an average length of 382 bp were generated from a full 454 GS-FLX run. After a trimming process removed adaptors, primer sequences, poly-A tails as well as short, long and low quality sequences, 822,891 (84.7%) high-quality reads were obtained with an average length of 358 bp covering a total of 21,076,176 bases (Table
Figure 1
Overview of
Overview of
The unigene coverage distribution revealed that most unigenes had a read-depth coverage <20-fold (Figure
Functional annotation of the transcriptome
The unigenes were annotated with gene names and Gene Ontology (GO) terms based on sequence comparisons between
Figure 2
Summary and taxonomic source of BLASTx matches to unigenes.
Summary and taxonomic source of BLASTx matches to unigenes. Number of unique best BLASTx matches of unigenes grouped by genus. The best matches of the unigenes to Pinaceae sequences accounted for 28.6% of the total.
Figure 3
Gene Ontology (GO) distributions for the
Gene Ontology (GO) distributions for the
Identification of SSRs, SNPs and Indels
Di- to hexa-nucleotide SSRs with a minimum repeat unit size of five (for tri- to hexa-nucleotide) or six (for di-nucleotide) were identified based on the analysis of assembled isotig templates. A total of 724 distinct loci were identified, and the incidences of different repeat types were determined. The tri-nucleotide repeats were most abundant (62.2%), followed by di-nucleotides (33.7%), among the various classes of SSRs (Figure
Figure 4
Distribution of simple sequence repeats (SSRs) in
Distribution of simple sequence repeats (SSRs) in
More than 92,000 SNPs/Indels were identified (61,454 SNPs and 31,030 Indels) from the
Figure 5
Quality of single nucleotide polymorphisms (SNPs) and insertion/deletions (InDels) in
Quality of single nucleotide polymorphisms (SNPs) and insertion/deletions (InDels) in
Orthologue identification and functional characterisation between six conifer species
Large-scale transcriptome characterisations have been carried out for
A comparative analysis of the transcriptomes of
Figure 6
Functional annotation and divergence between homologs of five pine and one spruce species.
Functional annotation and divergence between homologs of five pine and one spruce species. The heat map is based on the 191 putatively orthologous transcripts of six species. The homologs were annotated with Gene Ontology (GO) terms. Colours indicate similarity from yellow (highly similar) to red (weakly similar). The "2× average" is an overall measure of how similar the different species are.
Additional file 1
The 191 orthologues of six conifer species.
Click here for file
Phylogenetic and speciation analysis
Phylogenetic analyses of
Figure 7
Phylogram of the five pine and one spruce species.
Phylogram of the five pine and one spruce species. Phylogram derived using pairwise non-synonymous substitution rates of orthologous transcripts as a distance metric (not from multiple sequence alignments) and the neighbour-joining method
We estimated the level of synonymous substitutions for 191 pairs of orthologues identified among the six species and 6,053 pairs of orthologues identified between
Figure 8
Distribution of Ks values of orthologous pairs for identifying speciation events.
Distribution of Ks values of orthologous pairs for identifying speciation events. Data were grouped into bins of 0.02
Evolutionary pattern of
We estimated evolutionary measures at 6,053 orthologues of
Figure 9
Additional file 2
The 207 sequences with Ka/Ks ratios > 2 of
Click here for file
A large number of ESTs for pines have been sequenced to date (Table
Despite the fact that a large number of pine ESTs have been obtained from cDNA libraries based on traditional sequencing technology, the methods used were inefficient. Four
Next-generation sequencing technology yields a large number of sequences at considerably lower costs compared to traditional sequencing methods, and, therefore, provides a valuable starting point to expedite analysis of less-studied species
Comparative phylogenetic analysis at the genome level dramatically improves the precision and sensitivity of evolutionary inference
Understanding the factors that affect the evolutionary patterns and rates of genes is central in many research fields
This study is the first comprehensive sequencing effort and analysis of gene function in the transcriptome of
Sample collection, cDNA library creation and 454 sequencing
Total RNA isolation from samples of all selected plant tissues, and cDNA library construction and normalisation were performed as described previously
Assembly and annotation
All generated ESTs were pre-screened to remove adaptor-ligated regions and contaminants by Seqclean and to trim low-quality regions by LUCY2
The assembled isotig and singleton sequences were combined and clustered with CD-HIT (version 4.0)
We simultaneously instituted a search for putative unigenes against the NCBI protein database using a BLASTx and annotated each sequence with GO terms using Blast2GO.
Identification of SSRs, SNPs and InDels
Assembled isotigs with coverage of at least four reads were screened for SSRs, SNPs and InDels using Misa and ssahaSNP software, respectively
Identification of orthologues between six conifer species
The shared transcriptome data of five conifer species in the PlantGDB and NCBI databases were downloaded. The numbers of unigenes for each species were as follows:
Estimation at the level of synonymous substitution and non-synonymous substitution between orthologues
Because unigenes are derived from EST sequences, have no annotated open reading frames and may contain frame shift sequencing errors, each member of a pair of sequences was searched using BLASTX against all plant protein sequences available in GenBank. The approach used was as described previously
Phylogenetic analysis
Because the genus
Data availability
The raw 454 EST data obtained in this study were deposited in the NCBI Sequence Read Archive (SRA) under the accession number SRA 056887.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
SHN participated in the sequence alignment and drafted the manuscript. ZXL and HWY participated in the samples preparation and 454 sequencing. XYC and YL participated in the design of the study and performed the statistical analysis. WL conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.
This work was supported by grants from The Fundamental Research Funds for the Central Universities and the Special Fund for Scientific Forestry Research in the Public Interest (No.201104022).
Evolution of genome size in conifers
EuroPineDB: a high-coverage web database for maritime pine transcriptome
Expressed sequence tags from loblolly pine embryos reveal similarities with angiosperm embryogenesis
Stability in and correlation between factors influencing genetic quality of seed lots in seed orchard of
Phylogeography of
Cross-species transferability of SSR loci developed from transciptome sequencing in lodgepole pine
Conifer defence against insects: microarray gene expression profiling of Sitka spruce (
Combined proteomic and transcriptomic analysis identifies differentially expressed pathways associated to
Transcriptome sequencing in an ecologically important tree species: assembly, annotation, and marker discovery
Evolution of genome size and complexity in
Genomics to tree breeding and forest health
Gene discovery and annotation using LCM-454 transcriptome sequencing
Evaluating characteristics of de novo assembly software on 454 transcriptome data: a simulation approach
Large-scale transcriptome analysis in chickpea (
Sequencing and characterization of the guppy (
Identification, utilisation and mapping of novel transcriptome-based markers from blackcurrant (
Exploring the switchgrass transcriptome using second-generation sequencing technology
Transcriptome sequencing of field pea and faba bean for discovery and validation of SSR genetic markers
Mining transcriptome sequences towards identifying adaptive single nucleotide polymorphisms in Lake Whitefish species pairs (
SNP discovery via 454 transcriptome sequencing
Phylogeny, historical biogeography, and patterns of diversification for
Phylogeny and classification of
Colonization of the Tibetan Plateau by the homoploid hybrid pine
De novo characterization of the gametophyte transcriptome in bracken fern,
Generation and annotation of lodgepole pine and oleoresin-induced expressed sequences from the blue-stain fungus
Transcriptional analysis of
De novo sequencing and characterization of floral transcriptome in two species of buckwheat (Fagopyrum)
Slow but not low: genomic comparisons reveals slower evolutionary rate and higher
Generation and analysis of expressed sequence tags from six developing xylem libraries in
Transcriptome profiling of
Seasonal reorganization of the xylem transcriptome at different tree ages reveals novel insights into wood formation in Pinus radiata
Analysis of xylem formation in pine by cDNA sequencing
Apparent homology of expressed genes from wood-forming tissues of loblolly pine (
Large-scale statistical analysis of secondary xylem ESTs in pine
Water stress-responsive genes in loblolly pine (
Transcriptome sequencing of wild chickpea as a rich resource for marker development
Assembly, gene annotation and marker development using 454 floral transcriptome sequences in
Comparison of next generation sequencing technologies for transcriptome characterization
EST-SSRs as a resource for population genetic analyses
Cross-species transfer of nuclear microsatellite markers: potential and limitations
The molecular ecologist's guide to expressed sequence tags
Generation and analysis of blueberry transcriptome sequences from leaves, developing fruit, and flower buds from cold acclimation through deacclimation
Transcriptome characterization and high throughput SSRs and SNPs discovery in
Neighboring-nucleotide effects on single nucleotide polymorphisms: a study of 2.6 million polymorphisms across the human genome
Transition-transversion bias is not universal: a counter example from grasshopper pseudogenes
Evolution of genes and genomes on the
Comparative genomics based on massive parallel transcriptome sequencing reveals patterns of substitution and selection across 10 bird species
Transcriptome analysis and SNP development can resolve population differentiation of
A phylogenomic approach to resolve the arthropod tree of life
A 454 sequencing approach for large scale phylogenomic analysis of the common emperor scorpion (
EST sequencing of
An integrated view of protein evolution
The causes of protein evolutionary rate variation
The quest for the universals of protein evolution
Functional genomic analysis of the rates of protein evolution
LUCY2: an interactive DNA sequence quality trimming and vector removal tool
Comparing de novo assemblers for 454 transcriptome data
CD-HIT Suite: a web server for clustering and comparing biological sequences
Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences
Mining SNPs from DNA sequence data; computational approaches to SNP discovery and analysis
Novel tools for conservation genomics: comparing two high-throughput approaches for SNP discovery in the transcriptome of the European hake
Search and clustering orders of magnitude faster than BLAST
Widespread paleopolyploidy in model plant species inferred from age distributions of duplicate genes
PAML 4: phylogenetic analysis by maximum likelihood
MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0
Transcriptome-wide identification and characterization of miRNAs from


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A full review of the new Samsung Galaxy A5 Canadian version (also released in Europe). A new mid range phone with high end features from Samsung.
Samsung Galaxy A5 2017 Review Canada Model
Kangaroo Notebook Review
Want to keep your kids stuff separate from your personal stuff without having to buy two notebooks? InFocus has you covered with the new Kangaroo Notebook, it comes with TWO separate Kangaroo Mini's PC's that you dock into a Notebook! We review the new Kangaroo Notebook and let you know how it is!
$32 Kingmak Mini Full HD 1080p Car Dash Camera Review
Can a $32 dash cam really cut it or will it fall apart after a day? Watch as I review the Kingmak Mini Full HD 1080p Car Dash Cam!
You can buy it off Amazon US for $28 to $32, sometimes with a free micro sd card! Sorry, can't provide links as the seller keeps changing! Just search Amazon, Ebay or Alliexpress for: "Kingmak Mini Full HD 1080P Car Dash Camera" and make sure it looks like the one in my review.
Kangaroo Mobile Desktop Pro Review
InFocus sends us over a new Kangaroo Mobile Desktop Pro to review! This time the dock is loaded with features! We test out more gaming with Team Fortress 2, Serious Sam and more Mame gaming!
Thanks to inFocus for sending us a unit to review.
ChatSim Unlimited Review and Full Setup Step by Step
It's finally here ChatSim Unlimited has arrived! No more zones, it's just simply unlimited text chatting in WhatsApp, Telegram, BBM, Facebook Messenger, WeChat, QQi, Line, Hike and Kakao. We do an in depth review and show you how to setup ChatSim Unlimited.
Kangaroo Mini PC Plus Review – PSOne, Mame, League of Legends Gaming Tests
We review the new Kangaroo Mini PC Plus with double the ram and storage. We also answer your questions and test out PSOne, Mame, League of Legends Gaming!
Thanks to InFocus for the review unit!
iPhone SE Review / iPhone SE Benchmarks Versus iPhone 6s
In this video GearTactics host Justin Lee reviews the new iPhone SE. Is it really as fast as an iPhone 6s? We do iPhone SE versus iPhone 6s benchmarks and other tests to find out if it is!
D-Link Wireless AC3200 Tri-Band Gigabit DIR-890L Router Review
Your $90 router not cutting it because your have over 4 devices connecting to it? D-Link has you covered with the D-Link Wireless AC3200 Tri-Band Gigabit DIR-890L Router! We do an unboxing and review of this powerful router.
Thanks for D-Link for providing the product for us to provide the review.
Digiwave ANT 4500 HDTV Antenna Review and Unboxing
GearTactics reviews the Digiwave ANT 4500 flat antenna, sold for $15 by Best Buy Canada or Amazon in the United States…can it hold up to a $40 Channel Master or Silver Sensor indoor HDTV Antenna?
Find out by watching our review below:
This item was purchased for the review.
Kangaroo Mini PC by inFocus – Review and Unboxing
We review the Kangaroo Mini PC by inFocus. This is a $99 mini Windows 10 PC that can be used for surfing and office work. We decide to push our luck and do some gaming and MAME emulation with it also!
Available at NewEgg and the Microsoft Store Canada and USA. Thanks to inFocus for sending us the Kangaroo Mini PC for review unit!


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The latest collection of previously unavailable Conan adventures! The unstoppable Conan returns in another pile-driving selection of stories. These classic tales are newly available after more than a quarter of a century.
Bookwire is a mobile app for your iPhone that gives you the ability to scan a barcode
or enter ISBN, access book records and add titles to wish lists.


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Cleveland Outdoor Adventure Show Returns
January 11, 2017 The great outdoors is calling!! The Cleveland Outdoor Adventure Show is returning January 13-15th at the IX Center in Cleveland, Ohio. more »»
Sports Shows Slate is Full
January 11, 2017 Regional Calendar of Events 2017 Northeast Ohio Sportsman Show - Jan. 19-21, 2017 at new Mount Hope Event Center in Millersburg, OH. (Was formerly held at Buckeye Event Center in Dalton, OH. more »»
NE Ohio Sportsman Show to Move in 2017
December 19, 2016 MILLERSBURG, OH — Jody Witzky of JW Promotions, of Howard, OH, has announced that the 2017 Northeast Ohio Sportsman Show will be held at the new Mount Hope Event Center in Millersburg, OH on Januar... more »»
Great American Outdoor Show Opens February 6
February 5, 2016 HARRISBURG, PA — The 2016 Great American Outdoor Show, presented by Outdoor Channel, opens its doors February 6 at the Pennsylvania Farm Show Complex in Harrisburg, Pennsylvania. more »»
Ohio Decoy Carvers to Host Show
February 5, 2016 STRONGSVILLE, OH — The Ohio Decoy Carving and Collecting Association Show & Sale will be held March 12-13 in Strongsville (Cuyahoga County), OH. more »»
It's Show Time
January 26, 2016 Welcome to a new year, and new outdoor seasons. You may not know it's winter around here due to the wacky weather we've experienced so far; but nasty, cold and snowy days are looming. more »»
Great American Outdoor Show is Open in Harrisburg
February 9, 2015 HARRISBURG, PA – The 2015 Great American Outdoor Show, presented by Outdoor Channel, opened its doors on February 7. more »»
Great American Outdoor Show Comes to Close
January 31, 2014 HARRISBURG, PA – The Great American Outdoor Show closed out an impressive first show on February 9 at Pennsylvania Farm Show Complex & Expo Center in Harrisburg, Pennsylvania. more »»
Ohio Deer & Turkey Expo Nets Great Crowd
March 19, 2013 COLUMBUS, OH — A great crowd turned out for the 2013 Field and Stream Deer and Turkey Expo, held March 15-17 at the Ohio Expo Center in Columbus, OH. more »»
Another Great WV Show in Morgantown
March 6, 2013 MORGANTOWN, WV — Hats off to LaBeth Hall and the rest of her crew for putting on another great West Virginia Fishing, Hunting and Outdoor Show on March 2-3, 2013. more »»
New Friends Made at Allegheny Sports Show
February 19, 2013 MONROEVILLE, PA — Thanks to all of our subscribers — and new subscribers — who signed up at the latest Allegheny Sport, Travel and Outdoor Show in Monroeville, PA. more »»
Allegheny Sports, Travel and Outdoor Show Opens
February 14, 2013 MONROEVILLE, PA — An expected slow crowd kicked off the opening day of the Allegheny Sport, Travel and Outdoor Show on Wednesday in Monroeville, PA. more »»
News, Blogs & Events Web
The Great Outdoors
Ohio Parks: Visit Them For Free
July 12, 2017 I recently visited my daughter and her family, who live in the state of Indiana. more »
Latest Blogs


text:
|A League of His Own
|General Information
|Batman Gotham Adventures
|Issue Number:
|First Published:
|February 1999
|Previous Issue:
|Batman Gotham Adventures #8
|Next Issue:
|Batman Gotham Adventures #10
Batman Gotham Adventures #9: A League of His Own is the ninth issue of the Batman Gotham Adventures comic book series.
Continuing their pursuit of the League of Assassins, Batman and Batgirl trek to Nepal to assault the League's Himalayan stronghold. But two surprises are in store for the Dark Knight: the gun that Batgirl insists on carrying... and the identity of the League's leader.
Credits Edit
- Story: Ty Templeton
- Pencils: Rick Burchett
- Inks: Terry Beatty
- Letters: Tim Harkins
- Colors: Lee Loughridge
- Editor: Darren Vincenzo


text:
If I'm not working myself into a social media induced fervor, I'm walking around aimlessly downtown and listening to music.
Social Media-ing
Making up words and pretending it's Australian slang
Inserting Seinfeld quotes into any (read: every) conversation
Music: Hip Hop, Trap, Indie, other electronic-y stuff
Kindle - Gotta love them 'books'
Sun 'n' Beach - but I also have a semi-soft spot for the old rain
Funny stuff - if I'm not laughing at something or trying to make someone laugh then it means my day is pretty lame
Clothes - 💸
Dogs in NYC that wear better clothes than me
How to excel in life/work. I act all nonchalant when I really am, in fact, quite chalant.
- Know how to spell
- Love (update: tolerate. Updated update: TBD) Kanye
- Can maintain a conversation
- Understand humour/sarcasm


text:
Syndikut 5 Five Star Rating php script is a unique multiple rating system
Use this security logo:
Tutorial not found. Let know us about any useful video tutorial.
syndikut, script, scripts, multiple, scriptz, php, five, star, rate, rating, flat, file, demo, no sql, fsr, user, vote, system, flat file
New Release


text:
Real-imaging cDNA-AFLP transcript profiling of pancreatic cancer patients: Egr-1 as a potential key regulator of muscle cachexia
Cancer cachexia is a progressive wasting syndrome and the most prevalent characteristic of cancer in patients with advanced pancreatic adenocarcinoma. We hypothesize that genes expressed in wasted skeletal muscle of pancreatic cancer patients may determine the initiation and severity of cachexia syndrome.
Experimental design
We studied gene expression in skeletal muscle biopsies from pancreatic cancer patients with and without cachexia utilizing Real-Imaging cDNA-AFLP-based transcript profiling for genome-wide expression analysis.
Our approach yielded 183 cachexia-associated genes. Ontology analysis revealed characteristic changes for a number of genes involved in muscle contraction, actin cytoskeleton rearrangement, protein degradation, tissue hypoxia, immediate early response and acute-phase response.
We demonstrate that Real-Imaging cDNA-AFLP analysis is a robust method for high-throughput gene expression studies of cancer cachexia syndrome in patients with pancreatic cancer. According to quantitative RT-PCR validation, the expression levels of genes encoding the acute-phase proteins α-antitrypsin and fibrinogen α and the immediate early response genes Egr-1 and IER-5 were significantly elevated in the skeletal muscle of wasted patients. By immunohistochemical and Western immunoblotting analysis it was shown, that Egr-1 expression is significantly increased in patients with cachexia and cancer. This provides new evidence that chronic activation of systemic inflammatory response might be a common and unifying factor of muscle cachexia.
Pancreatic cancer remains one of the most deadly tumor types, with the highest death-to-incidence ratio of all cancers. With a 5-year survival rate of less than 5% and a death-to-incidence ratio of 0.99, pancreatic ductal adenocarcinoma is currently one of the most aggressive gastrointestinal carcinomas
In the case of muscle wasting associated with cancer, the balance between protein synthesis and degradation is shifted markedly toward degradation, and there could be selective targeting of skeletal muscle gene products by different cachectic factors such as cytokines and tumor-derived compounds
Unfortunately, patients with pancreatic cancer generally present in late stage of the disease, and frequently develop cachexia syndrome. This emphasizes the importance of early diagnosis, especially with the hope that intervention may be more effective in the early stages of disease. This also underscores the need for biomarkers, which may also contribute to the understanding of cancer cachexia syndrome
Here we present our study that focuses on the identification of common differentially expressed genes involved in muscle wasting progression. We hypothesized that gene expression patterns in the skeletal muscle of cancer patients will change under muscle wasting conditions, and that these changes may determine the initiation and severity of cachexia syndrome. We suggest that characterizing the skeletal muscle in response to cancer cachexia will provide new insights into gene regulation.
Patients with pancreatic cancer were selected for this study, as these patients usually experience severe weight loss (weight loss exceeding 10% of the stable body weight). All included patients gave written informed consent for tissue and data collection under the local ethics committee's permission of the University of Heidelberg.
On admission to hospital, each patient was asked for his height and current weight. When a patient gave a history of weight loss, the corresponding time period was also documented. A patient was classified as cachectic when weight loss exceeded 10% of the stable pre-illness weight. It has been shown, that in patients with a history of weight loss exceeding 10% of the stable weight, the survival is significantly reduced
Muscle biopsies
Biopsy specimens were obtained from the rectus abdominis muscle of each patient during the initial phase of the operation. After skin incision and dissection through the subcutaneous fat, the anterior sheet of the rectus abdominis muscle was opened with scissors and a muscle biopsy specimen weighing about 50 mg was obtained and immediately frozen in liquid nitrogen.
Real-Imaging cDNA-AFLP transcript profiling
mRNA was extracted from 20 mg rectus abdominis muscle using MagNA pure LC isolation system according to manufacturer's instructions (Roche). After extraction, samples were immediately stored at −80°C until usage. mRNA quality was assessed by reverse transcription PCR (RT-PCR) with two housekeeping genes encoding ribosomal protein L13a (RPL13A) and β-actin. Negative controls for genomic contamination and reverse transcription inhibition were included.
The cDNA-AFLP procedure was performed as described previously with some modifications
cDNA-AFLP amplification was performed with 16 primer pairs MseI+2 and TaqI+2IRD700 (+2 states for additional two-nucleotide combination at 3'-end of primer) generating approximately 30,000 amplified fragments possessing 256 probe sets. IRD-labeled PCR products were resolved in 40 cm long 6% SequaGel XR gel (Biozym) using automated DNA sequencer DNAIR 4200 (Li-COR). After 1 h electrophoresis the gel was transferred onto the green grid paper and vacuum dried on 1.2-mm-thick Whatman paper. The dried gel was scanned at 700/800 nm (intensity 4.0, resolution 84 μm), obtaining binary data for differentially expressed tags (Odyssey Imaging System, Li-COR).
Isolation and analysis of cDNA-AFLP tag fragments
The cDNA fragments of interest were excised from dried gel, followed by elution in 100 μl TE for 10 min at 95°C. 5 μl were re-amplified with TaqI-Ø and MseI-Ø primers. Specific fragments were verified by visualizing 5 μl PCR product on a 1.5% agarose gel. The amplified fragments were directly sequenced with the primers TaqI-ØIRD700 and MseI-ØIRD800 using the SequiTherm EXCEL II DNA sequencing kit (Epicentre) on the automated infrared sequencer DNAIR 4200 (Li-COR).
Sequencing data were aligned against BlastX and BlastN in the publicly available databases and clustered according to gene ontology using the nodes of the DNASpace software package (DNASIS MAX 2.6, Miraibio).
Quantitative RT-PCR
Single-stranded cDNA was synthesized from 100 ng of mRNA according to the manufacturer's instructions (RevertAid, Fermentas). Primers complementary to target genes were designed using Primer Express Software (Life Technologies) to amplify fragments in the range from 75 bp to 200 bp, close to the 3'-end of transcripts. A quantitative RT-PCR (qPCR) was performed with SYBR Green PCR Master Mix using GeneAmp 7300 (Life Technologies). qPCR reactions were performed two times in doublets, and the threshold cycle numbers were averaged. Levels of expression were normalized against RPL13A and β-actin.
For immunohistochemistry, 5-μm sections of FFPE tissue were incubated in serum-free protein blocking buffer for 30 min, followed by overnight incubation at 4°C with 1:200 dilution of Egr-1 antibodies (588, Santa Cruz). A negative control for non-specific staining was performed with rabbit IgG. DAKO anti-rabbit polymer-HRPO was applied for signal visualization according to the manufacturer's instructions (DAKO). Finally, sections were analyzed with an Olympus BX50 microscope using 40x-lens and software program "Cell^F 2.5" (Olympus).
Protein extraction and immunoblotting
30 μg total protein lysate from the biopsy homogenate were electrophoresed on gradient 4-15% SDS-PAA gels (Bio-RAD). After undergoing electrophoresis, proteins were transferred semi-dried onto PVDF membrane using the Semi-Dry Blotter (Bio-RAD). After 1 h blocking, the membrane was exposed overnight at 4°C to primary anti-human rabbit Egr-1 antibody (588, Santa Cruz) and anti-human mouse β-tubulin antibody (3 F3-G2, Santa Cruz) at 1:200.
After washing the membrane was exposed to appropriate IRDye-700/800 labeled secondary antibodies: Alexa Fluor 680 goat anti-mouse for β-tubulin (Invitrogen) and IRDye-800 goat anti-rabbit for Egr-1 (Rockland) at 1:10,000. The amounts of Egr-1 and β-tubulin were determined and quantified by densitometric scanning of PVDF membranes at 700/800 nm (intensity 5.0, resolution 84 μm) using the ODYSSEY Infrared Imaging System (Li-COR). Individual band intensities were normalized against β-tubulin.
Statistical analysis
All data are presented as means ± standard deviation. The significance of differences between groups (cachexia vs. no cachexia) was tested with Student's
Patient characteristics
We studied skeletal muscle biopsy samples of 23 patients diagnosed with histologically proven adenocarcinoma of the pancreas and treated at the Department of Surgery, University of Heidelberg, between November 2004 and April 2005. In ten of the patients weight loss exceeded 10% of the stable pre illness weight, while weight loss was absent or less than 10% of the stable weight in 13 patients. Clinical characteristics of these patients are summarized in Table
Initial staging of these patients was determined according to the 5th edition of the TNM classification of the International Union against Cancer. UICC classification of operated and examined patients (N = 23) is shown in Figure
Figure 1
UICC Classification of the 23 patients operated for pancreatic cancer
UICC Classification of the 23 patients operated for pancreatic cancer. Distribution of tumor stage of all pancreatic cancer patients was determined according to the 5th edition of the TNM classification of the Union International Contre Cancer (UICC). Median weight loss of the cachectic patients was 13.9% of initial body weight within 6 months comparing with the patients without cachexia in which the median weight loss was 2.1% respectively. The resection rate was lower in patients with cachexia (50% vs. 92.3%) UICC II: without distant metastases; UICC III: infiltration of celiac trunk and/or mesenteric artery; UICC IV: metastases in liver and/or peritoneal cavity according to guidelines UICC 2002) in patients either with (n = 10) or without cachexia (n = 13).
Real-Imaging cDNA-AFLP screening
We performed gene expression profile screening of the skeletal muscle of pancreatic cancer patients either with or without cachexia syndrome utilizing Real-Imaging cDNA-AFLP transcript profiling. Biopsy samples from 10 wasted patients and 13 patients without cachexia were compared by running the cDNA-AFLP reaction using a single primer combination. This comparison revealed that amplified tag patterns are almost identical within one group and differ between the groups (Additional file 1: Figure S1). Therefore, two biopsy samples from each group were further selected for preparative Real-Imaging cDNA-AFLP (Figure
Additional file 1
Figure S1.cDNA-AFLP: Comparison of 10 cachectic and 13 non-cachectic patients using one-primer-pair combination.
Click here for file
Figure 2
cDNA-AFLP Transcript Profiling of pancreatic cancer patients
cDNA-AFLP Transcript Profiling of pancreatic cancer patients. cDNA-AFLP tag patterns amplified from mRNA of skeletal muscle biopsy samples of pancreatic cancer patients either with cachexia (IRD800-labeled, red bands) or without cachexia (IRD700-labeled, green bands) were loaded in the same lane according to their primer combinations and separated in a 8% denaturing polyacrylamide gel. Data were collected on automated Infrared Sequencer DNAIR 4200 (Li-COR Biosciences GmbH). Three different cDNA-AFLP tag profiles (i), (ii) and (iii) of wasted patients (red bands) were compared with one non-cachectic control (green bands).
Altered expression patterns were readily recognized by visual inspection and yielded approximately 400 cDNA fragments (Figure
Figure 3
Preparative Real-Imaging cDNA-AFLP Screening
Preparative Real-Imaging cDNA-AFLP Screening.A) cDNA tag patterns were amplified with individual primer combinations Taq+2 and Mse+2 (Taq+2 was labeled with IRD700). After electrophoresis, the gel was transferred and dried on green grid paper and scanned using Odyssey Imaging System (Li-COR Biosciences GmbH). The lanes are occupied by cachexia-associated samples (+) and non-cachectic probes (−) allowing direct visual comparison. Grid coordinates are indicated in millimeters (mm). B) Differentially expressed tag patterns (marked in red boxes) were isolated for sequencing analysis.
Analysis of differentially expressed genes
183 cDNA sequences were identified after a redundancy check of about 400 isolated tag fragments. They were clustered into groups according to their annotation fields. 127 cDNA fragments were referred to the gene transcripts that predominantly activated in cachectic patients, as well as 56 individual genes whose transcripts accumulated predominantly in the non-cachectic control group (Additional file 2: Table S1).
According to group classification, the identified genes provide molecular counterparts of muscle cachexia responsible for actin cytoskeleton rearrangement, actin filament disruption, extracellular matrix dysregulation, protein degradation, energy expenditure, tissue hypoxia, immediate early response, and acute-phase response. We identified a number of differentially expressed genes, which have already been characterized in relation to cancer cachexia or muscle atrophy. However, our results also indicate that there are other differentially expressed genes, which play unknown roles in muscle cachexia.
In wasted patients we observed the upregulation in gene expression of the contractive muscle proteins (Additional file 2: Table S1), correlating directly with other findings
Additional file 2
Table S1.Candidate biomarkers for cachexia progression. Legend: diff. - differential gene expression: "+" cachexiaspecific upregulation,"-"cachexia-specific downregulation.
Click here for file
Validation of cachexia-associated genes with qPCR
The differential gene expression level of 100 transcripts from cDNA-AFLP screening profile corresponding to the genes, which are associated with protein degradation, hypoxia, acute-phase response or with disorder in muscle tissue development, was validated by quantitative RT-PCR of the same skeletal biopsy samples (n = 23) using a Mann–Whitney
Egr-1 protein level is highly elevated in wasted skeletal muscle
To prove our cDNA-AFLP gene profiling strategy, followed by qPCR validation, the protein expression levels of transcription factor Egr-1 were further analyzed by immunostaining and Western immunoblotting. The expression sites of Egr-1 protein were identified both in longitudinal and transverse sections of the skeletal muscle from patients either with or without cachexia by immunohistochemical staining with Egr-1 antibodies. Comparison of tissue sections revealed a significantly stronger immunohistochemical signal of Egr-1 protein in the skeletal muscle tissue cells of patients with cachexia, as indicated by increased fluorescent staining (Figure
Figure 4
Altered expression of Egr-1 in the skeletal muscle tissue of wasted pancreatic cancer patients
Altered expression of Egr-1 in the skeletal muscle tissue of wasted pancreatic cancer patients. Immunohistochemical staining of skeletal muscle tissue shows significantly enhanced expression of Egr-1 in wasted pancreatic cancer patients (C, D) compared with non-cachectic patients (A, B). A, C – longitudinal sections; B, D – transverse sections of skeletal muscle from pancreatic cancer patients. 40X original magnification.
Furthermore, Egr-1 protein (82 kDa) was quantified in protein extracts from muscle biopsy samples by Western immunoblotting using densitometric quantitation of the appropriate bands against β-tubulin by infrared fluorescence detection directly on the PVDF membrane (Figure
Figure 5
Egr-1 protein is markedly induced in skeletal muscle of wasted pancreatic cancer patients
Egr-1 protein is markedly induced in skeletal muscle of wasted pancreatic cancer patients. Protein levels of skeletal muscle EGR-1 and β-tubulin (loading control) were determined by Western immunoblotting showing aberrant upregulation of Egr-1 in skeletal muscle of patients with cancer cachexia. Western immunoblotting of Egr-1 (green band, 82 kDa) and β-tubulin (red band, 55 kDa) compares (A) two non-cachectic with two cachexia-associated biopsy protein extracts and (B) one non-cachectic probe with three cachexia-associated skeletal muscle biopsy samples. Similar results were obtained in three independent experiments. M – coomassie-blue prestained protein marker. Full-length blots/gels are presented in Additional file 3: Figure S2. (C) Densitometric analysis of Egr-1 protein expression. Median rate = 10.23, P-value ≤ 0.005 according to Student's
Finally, immunohistochemistry and Western immunoblotting analysis confirmed the strong correlation of Egr-1 within the skeletal muscle of pancreatic cancer patients in response to cachexia.
Additional file 3
Figure S2.Full-length Western blotting of Egr-1.
Click here for file
Cachexia is present in around 80% of patients with cancer in advanced stages, inducing marked depletion of lean and fat mass, and in more than 20% of patients cachexia is responsible for death
We hypothesized that alteration in gene expression within the skeletal muscle in response to advanced pancreatic cancer plays a crucial role in the development of the muscle cachexia syndrome. To test this hypothesis, we carried out the transcriptome-wide characterization of skeletal muscle biopsies utilizing the Real-Imaging cDNA-AFLP approach to identify those genes that are commonly induced in the skeletal muscle of pancreatic cancer patients either with or without muscle cachexia.
In contrast to other studies of skeletal muscle cancer cachexia based on cDNA microarray technique
We observed that transcript levels of contractile proteins in skeletal muscle of patients with cachexia change in the same direction. We observed more than 6-fold upregulation of cyclin-dependent kinase inhibitor p21 (Table
We have also observed the induction of genes encoding basic helix-loop-helix (bHLH) transcription factors HES1, DEC1, and DEC2, which are primarily associated with hypoxia (Table
Our study demonstrates for the first time that the genes encoding acute-phase response (APR) proteins fibrinogen α, α1-antitrypsin and α2-macroglobulin are markedly upregulated not only in the liver but also in the skeletal muscle of wasted patients with pancreatic cancer (Table
While acute-phase response indicates systemic inflammation, the pro-inflammatory cytokines appear to mediate this response. Furthermore, it has been reported that non-immune tissues such as cardiac and skeletal muscle appears to express pro-inflammatory cytokines
Notably, in contrast to the previous study on human skeletal muscle cancer cachexia
Importantly, together with an ongoing inflammatory response we observed the significant induction of early response genes Egr-1 and IER-5 in wasted pancreatic cancer patients (p<0.05, Table
This report provides the first evidence that Egr-1 is aberrantly upregulated under conditions of muscle wasting, underlining the general importance of pro-inflammatory response in cancer cachexia (Figures
In summary, taken together with previous works in the field, the current study supports a model of muscle wasting in which the major influence of cancer cachexia is based on the chronic inflammatory effects resulting in onset of acute-phase response. Furthermore, this report provides new insights into the inflammatory response involved in initiation and progression of cachexia by the activation of pro-inflammatory transcription factor Egr-1 and acute-phase response. How they are transduced to initiate the muscle cachexia remains to be investigated.
cDNA-AFLP, cDNA amplified fragment-length polymorphism; Egr-1, early growth response; 1IER-5: immediate early response 5; APR, Acute-phase response.
Competing interests
All authors have completed the ICMJE uniform disclosure form at
J. Bachmann, N. Giese and M. Martignoni declare: no support from any organisation for the submitted work; no financial relationships with any organisations that might have an interest in the submitted work in the previous 3 years; no other relationships or activities that could appear to have influenced the submitted work.
Authors' contributions
Every author was involved in conception and design. AS carried out both molecular genetic studies and protein expression analysis. AS and JB drafted the manuscript. AS, JB, MM, HKR participated in analysis and interpretation of data.MM, NG, HKR revised it critically for important intellectual content and final approval of the version to be published. All authors read and approved the final manuscript.
We thank Brunhilde Bentzinger and Inna Babushkina for their technical assistance in setting up the immunohistochemical experiments.
This work was supported by German Cancer Research Center.
Grant funding for research
A. Skorokhod and H. Krakowski-Roosen declare:
These two authors had financial support from the German Cancer Research Centre for the submitted work; no financial relationships with any organisations that might have an interest in the submitted work in the previous 3 years; no other relationships or activities that could appear to have influenced the submitted work.
Cancer statistics, 2009
Cachexia worsens prognosis in patients with resectable pancreatic cancer
Rapid disuse and denervation atrophy involve transcriptional changes similar to those of muscle wasting during systemic diseases
Molecular mechanisms involved in muscle wasting in cancer and ageing: cachexia versus sarcopenia
Systemic inflammation, cachexia and prognosis in patients with cancer
Pro-inflammatory cytokines and oxidative stress/antioxidant parameters characterize the bio-humoral profile of early cachexia in lung cancer patients
Pancreatic cancer related cachexia: influence on metabolism and correlation to weight loss and pulmonary function
2010 TNM system: on the 7th edition of TNM classification of malignant tumors
cDNA-AFLP analysis of differential gene expression in the prokaryotic plant pathogen Erwiniacarotovora
Restoration of hormonal action and muscle protein
Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy
Cachexia and obesity: two sides of one coin?
Hypercatabolism and hypermetabolism in wasting states
Using transcriptomics to identify and validate novel biomarkers of human skeletal muscle cancer cachexia
Transcriptional profile of a myotube starvation model of atrophy
The possible role of myostatin in skeletal muscle atrophy and cachexia
Role of mononuclear cells and inflammatory cytokines in pancreatic cancer-related cachexia
Hypoxia requires notch signaling to maintain the undifferentiated cell state
Impaired expression of Notch signaling genes in aged human skeletal muscle
Pro-inflammatory cytokine release by peripheral blood mononuclear cells from patients with advanced pancreatic cancer: Relationship to acute phase response and survival
Definition of cancer cachexia: effect of weight loss, reduced food intake, and systemic inflammation on functional status and prognosis
Skeletal muscle: a dual system to measure glucocorticoid-dependent transactivation and transrepression of gene regulation
IL-32, a novel cytokine with a possible role in disease
Proinflammatory role of leukocyte-derived Egr-1 in the development of murine postoperative ileus
Egr-1, a master switch coordinating upregulation of divergent gene families underlying ischemic stress
Cytokines and their role in the pathophysiology of cancer cachexia
Pre-publication history
The pre-publication history for this paper can be accessed here:


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NEW YORK ( TheStreet) -- The massive offering of MetLife ( MET) shares related to the company's purchase of AIG's ( AIG) Alico business priced at $43.25 per share late Wednesday. The deal involved a total of 146.8 million MetLife common shares. AIG is selling 78.2 million of the shares, while MetLife is offering the other 68.6 million and plans to use its proceeds to purchase and cancel preferred stock held by AIG from the ALICO deal.
The Treasury Department issued a statement on Wednesday saying the expected $6.3 billion in gross proceeds from the offering to go towards AIG's bailout tab, which currently stands at $18.2 billion. "This is the next chapter in AIG's remarkable turnaround," said Acting Assistant Secretary for Financial Stability Tim Massad said in the statement. "We are optimistic about the prospects that taxpayers will recover every dollar invested in AIG - something that many thought would be impossible when these investments were first made." AIG also priced a public offering of 40 million MetLife common equity units at $75 each. The $3 billion in proceeds from this offering are to go into an escrow account to pay any obligations that AIG may end up owing MetLife under the terms of the ALICO sale. After a holding period of two years, the remaining proceeds would then be released to the Treasury. MetLife shares closed Wednesday's regular session at $43.41, down 5.7%, on more than five times its usual daily volume, while AIG's stock rose 1.6% to $37.30. -- Written by Michael Baron in New York. >To contact the writer of this article, click here: Michael Baron. >To submit a news tip, send an email to: email@example.com


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The core foundation of Hindu belief is that Vedas contain source of all knowledge – physical or metaphysical. Vedas and Puranas, which were revealed more than 5,000 years ago, mentioned facts only recently discovered or proven by scientists.
For a quick glimpse at what unsung surprises may lie in the Vedas, let us consider these renditions from the Yajur-veda and Atharva-veda, for instance.
" O disciple, a student in the science of government, sail in oceans in steamers, fly in the air in airplanes, know God the creator through the Vedas, control thy breath through yoga, through astronomy know the functions of day and night, know all the Vedas, Rig, Yajur, Sama and Atharva, by means of their constituent parts."
" Through astronomy, geography, and geology, go thou to all the different countries of the world under the sun. Mayest thou attain through good preaching to statesmanship and artisanship, through medical science obtain knowledge of all medicinal plants, through hydrostatics learn the different uses of water, through electricity understand the working of ever lustrous lightening. Carry out my instructions willingly." (Yajur-veda 6.21).
" O royal skilled engineer, construct sea-boats, propelled on water by our experts, and airplanes, moving and flying upward, after the clouds that reside in the mid-region, that fly as the boats move on the sea, that fly high over and below the watery clouds. Be thou, thereby, prosperous in this world created by the Omnipresent God, and flier in both air and lightning." (Yajur-veda 10.19).
" The atomic energy fissions the ninety-nine elements, covering its path by the bombardments of neutrons without let or hindrance. Desirous of stalking the head, ie. The chief part of the swift power, hidden in the mass of molecular adjustments of the elements, this atomic energy approaches it in the very act of fissioning it by the above-noted bombardment. Herein, verily the scientists know the similar hidden striking force of the rays of the sun working in the orbit of the moon." (Atharva-veda 20.41.1-3).
Some of the scientific statements as written in Vedas are :
Rig Veda 8.12.28
"O Indra! by putting forth your mighty rays, which possess the qualities of gravitation and attraction-illumination and motion – keep up the entire universe in order through the Power of your attraction."
Rig Veda 1.6.5, Rig Veda 8.12.30
"O God, You have created this Sun. You possess infinite power. You are upholding the sun and other spheres and render them steadfast by your power of attraction."
Rig Veda 5.40.5
"O Sun! When you are blocked by the one whom you gifted your own light (moon), then earth gets scared by sudden darkness."
Rig Veda 1.84.15
"The moving moon always receives a ray of light from sun"
Rig Veda 10.22.14
"This earth is devoid of hands and legs, yet it moves ahead. All the objects over the earth also move with it. It moves around the sun.
The existence of rather advanced concepts like the sphericity of Earth and the cause of seasons is quite clear in Vedic literature. For example, the Aitareya Brahmana (3.44) declares:
The Sun does never set nor rise. When people think the Sun is setting it is not so. For after having arrived at the end of the day it makes itself produce two opposite effects, making night to what is below and day to what is on the other side.Having reached the end of the night, it makes itself produce two opposite effects, making day to what is below and night to what is on the other side. In fact, the Sun never sets.
'Shape of Earth is like an Oblate Spheroid. (Rig Veda 30.4.5)
'Earth is flattened at the poles' (Markandeya Purana 54.12)
"Blue Sky is Nothing but scattered sunlight" (Markandeya Purana 78.8)
Now lets see what Modern Science says about it:
The blue color of the sky is due to Rayleigh scattering. As light moves through the atmosphere, most of the longer wavelengths pass straight through. Little of the red, orange and yellow light is affected by the air.
"To the philosophers of India , Relativity is no new discovery, just as the concept of light years is no matter for astonishment to people used to thinking of time in millions of kalpas, (A kalpa is about 4,320,000 years). The fact that the wise men of India have not been concerned with technological applications of this knowledge arises from the circumstance that technology is but one of innumerable ways of applying it."
It is, indeed, a remarkable circumstance that when Western civilization discovers Relativity it applies it to the manufacture of atom-bombs, whereas Oriental civilization applies it to the development of new states of consciousness."
(source: Spiritual Practices of India – By Frederic Spiegelberg Introduction by Alan Watts p. 8-9).
The Indians came closest to modern ideas of atomism, quantum physics, and other current theories. India developed very early, enduring atomist theories of matter. Possibly Greek atomistic thought was influenced by India, via the Persian civilization. The Rig-Veda, is the first Indian literature to set down ideas resembling universal natural laws. Cosmic law is connected with cosmic light, with gods, and, later, specifically with Brahman. It was the Vedic Aryans who gave the world some of the earliest philosophical texts on the makeup of matter and the theoretical underpinnings for the chemical makeup of minerals. Sanskrit Vedas from thousands of years before Christ implied that matter could not be created, and that the universe had created itself. Reflecting this, in his Vaiseshika philosophy, Kanada (600 B. C) claimed that elements could not be destroyed. Kanada's life is somewhat a mysterious, but his name is said to mean "one who eats particle or grain" likely referring to his theory that basic particles mix together as the building blocks for all matter. Two, three, four, or more of these elements would combine, just as we conceive of atoms doing. The Greeks would not stumble on this concept for another century."
Renowned Thinkers Who Valued the Knowledge in Vedas
Arthur Schopenhauer, the famed German philosopher and writer, wrote that: "I encounter [in the Vedas] deep, original, lofty thoughts… suffused with a high and holy seriousness."
The well-known early American writer Ralph Waldo Emerson, read the Vedas daily. Emerson wrote: "I owed a magnificent day to the Bhagavat-Gita"
Henry David Thoreau said: "In the morning I bathe my intellect in the stupendous philosophy of the Bhagavad Gita… in comparison with which… our modern world and its literature seems puny and trivial."
So great were Emerson and Thoreau's appreciation of Vedantic literatures that they became known as the American transcendentalists. Their writings contain many thoughts from Vedic Philosophy.
Another famous personality who spoke of the greatness of the Vedas were: Alfred North Whitehead (British mathematician, logician and philosopher), who stated that: "Vedanta is the most impressive metaphysics the human mind has conceived."
Julius Robert Oppenheimer, the principle developer of the atomic bomb, stated that "The Vedas are the greatest privilege of this century." During the explosion of the first atomic bomb, Oppenheimer quoted several Bhagavad-gita verses from the 11th chapter, such as:
"Death I am, cause of destruction of the worlds…"
When Oppenheimer was asked if this is the first nuclear explosion, he significantly replied: "Yes, in modern times," implying that ancient nuclear explosions may have previously occurred.
Friedrich Maximilian Miller (1823-1900) was a German philologist and Orientalist. He repeatedly drew attention to the uniqueness of the Vedas and awakened interest in his book In History of Ancient Sanskrit Literature' (p. 557) observed:
" In the Rig-Veda we shall have before us more real antiquity than in all the inscriptions of Egypt or Ninevah….the Veda is the oldest book in existence…."
The late scientist, Carl Sagan, in his book, Cosmos asserts that the Dance of Nataraja, Tandava signifies the cycle of evolution and destruction of the cosmic universe (Big Bang Theory).
"It is the clearest image of the activity of God which any art or religion can boast of." Modern physics has shown that the rhythm of creation and destruction is not only manifest in the turn of the seasons and in the birth and death of all living creatures, but also the very essence of inorganic matter.
The above are just few examples showing the scientific foreknowledge of Vedas and Puranas. Vedas are treasures of wisdom which are not yet fully explored.
Hindu tradition sometimes holds that all knowledge is pre-existing, to be "recovered" rather than "discovered", and is echoed can be found in the Vedas and other ancient texts.
In 1900, Vivekananda said:
"It seems to us, and to all who care to know, that the conclusions of modern science are the very conclusions the Vedanta reached ages ago; only, in modern science they are written in the language of matter."


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A Lower Mount Bethel Township man is accused of forging a Nazareth doctor's name in an attempt to obtain oxycodone from the Wal-Mart pharmacy in Lower Nazareth Township.
Chad Wagner, 23, of the 6000 block of Franklin Hill Road, was arraigned before District Judge John Capobianco and charged with forgery, criminal attempt and criminal trespass — all felonies. He was also charged with theft, receiving stolen property and prohibited acts; penalties.
He is free on $15,000 unsecured bail.
According to a police criminal complaint:
On Tuesday, May 22, Officer Stephen Schleig responded to Dr. Sabre's office at 111 S. Spruce St. for a forgery report.
Sabre said he was notified by the Wal-Mart pharmacy that an unknown male had submitted a forged prescription for 60 oxycodone pills. According to Sabre, the prescription was in an employee's name, Pamula Plummer, who was present in his office at the time.
Sabre told Schleig he believed the unknown male was Wagner, Plummer's boyfriend.
Plummer confirmed that she was in a relationship with Wagner, and said he has been in and out of rehab for heroin and oxycodone abuse.
Wagner confessed that he forged Sabre's signature on a prescription note he had stolen from inside a locked cabinet located inside Plummer's office. He was able to do this, he said, when Plummer stepped out to use the restroom.
Read More:
Police: Man Forged Nazareth Doctor's Signature for Oxycodone – Patch.com


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Banned: U.S. Trade Sanctions
In a world of increasingly free trade, the U.S. routinely imposes its own trade sanctions on many nations. Typically they are for reasons relating to human rights abuse and other hostile gestures. The more prominent of these sanctions are described below.
Belarus: Political Repression, 06/2006
Prohibits donations to anyone undermining the democratic process in Belarus.
Balkan Nationals: Extremist Violence, 06/2001
Prohibits donations to people participating in or supporting violent behaviors.
Iran: Supporting International Terrorism, 1987 and 2000
No goods may be imported to the U.S., other than gifts under $100 in value, informational materials, food and carpets.
Iraq: Kuwait Invasion, 1990 and 2004
Exports must be licensed by the Department of Commerce. It prohibits transactions with respect to Iraqi cultural property.
North Korea: Proliferating Weapons, 06/2008
The ban prohibits importing any good that has not been approved by the U.S. government. It bans any U.S. citizen from registering a vessel in North Korea.
Burma/Myanmar: Reduce Military Funding, 06/2008
The ban prohibits the importing of most goods from Burma.
Zimbabwe Nationals: Undermining Democracy, 11/2005
The ban prohibits all U.S. citizens from engaging in any transactions with anyone who is undermining the democratic process.
D.R. Congo: Increase Regional Instability, 10/2006
Prohibits making contributions or donating to anyone participating in the conflict.
Cote D'Ivoire: Human Rights Violations, 02/2006
The ban prohibits any U.S. citizen from engaging with any person involved with conflict.
Syria: Human Rights, 02/2006
The ban prohibits any U.S. citizen from engaging with any person involved with conflict.
Cuba: Response to Hostile Actions, 07/1963
No goods can be imported from Cuba. With a few exceptions, only people with a close blood relative may visit Cuba.


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Bath Salts Could Cause Flesh-Eating Bacteria
A 34 year-old New Orleans woman lost her arm this summer after shooting bath salts. She originally went to the hospital for pain and swelling in her forearm.
The doctor diagnosed a skin infection and put her on intravenous antibiotics. The symptoms improved, at first, but then her condition worsened.
She admitted that she had intravenously shot bath salts several days earlier. Doctors discovered a flesh-eating bacteria under her skin. The bacteria spread so quickly that doctors could see healthy tissue dying right before their eyes.
In order to save her life, doctors amputated her arm and shoulder. The doctor in charge of the case warns emergency room doctors that bath salts could be the cause of this flesh-eating bacteria. Read more here. And read a more scientific story in a medical journal.
Picture courtesy of OrthoSuperSite.


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Tommy shouts back: "What do you mean it's too loud?"
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