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2017 Artisan Directory
Here's a full list of the artisans who attended Fair December 2017. The vendor list for 2018 will be updated closer to the time.
Click on the artisan's name below for more information.
Chic Made Consciously
Chic made consciously offers fashionable accessories made sustainably, we work with artisans in Bali Indonesia who handcraft our line of accessoriesmade from repurposed tire innertubes. Our mission is to encourage conscious shopping by promoting fair trade and reduction of waste.
Handmade poly/spandex headbands and neckwarmers, for yoga, running, skiing, walking or just wearing to the store. Beautiful color selections – something for everyone on your list.
HML Designs
Millinery: hand blocked sinamay and wool felt fascinators and silk hair accessories for special occassions.
Conscious Artt
I create Orgonite Sculptures. These pyramids are a mixture of catalyzed resin with metal shavings and healing crystals poured into molds. Orgonite draws in negative life energy and transmutes it into positive energy.
Leslie Buckle
Prints and oil paintings by local Port Credit artist Leslie Buckle.
Nancy W. Phillips. Visual Artist
Nancy primarily uses watercolor and acrylic as her medium. Her main emphasis is landscapes.
Robin Hollingdrake
Set-Apart Creations
Sue Archibald Artist
Original acrylic paintings and small giclee prints on canvas.
Captain Jamie's Holistic Body Care Products
Holistic body care products, medicinal balms and creams. Absolutely natural and healing. Choose a better way. Choose Captain Jamie's www.captainjamies.com .
Nature's Purebliss
Skin and haircare therapy, body mousse, oils, gourmet tub cookies and chocolate scrubs. Visit their website at www.naturespurebliss.com
Terrah Essentials
A local Port Credit busing sharing natural skin care products with those looking for a safer more natural option to look and feel good.
Wenona Lake
Handcrafted Glycerin Soap
Across The Miles
Handmade alpaca hats, scarves and rugs. Natural stone jeweler, hand painted boxes, natural product Christmas ornaments.
Caryl Richmond
Clothing and accessories created from upcycled and reclaimed textiles.
Designed To Life
One-of-a-kind and unique designs for t-shirts, onesies, tote bags, post cards and framed prints to inspire positive change in the person and the world around them. Visit their website at designedtolife.com
Fibre Artists
Weavers of cotton tea towels, guest towels, runners, place mats, woven silk, wool & chenille scarves, throws and stoles in a variety of yarns and some beautiful hand painted scarves.
Locally Sewn Textiles
The philosophy behind this shop is to provide unique handmade scarves using quality materials with attention to originality and craftsmanship
Lovebird Design
Lovebird Tee's is both a response to a need for beauty, vibrant color and art in our every day lives. From the unique quality of the hand drawn images, to the organic cotton and fair wage manufacture of its tees, Lovebird strives to ensure that the elements we create are "earth and people kind."
Luna Sea
Handmade clothing – ponchos, wraps, hats, – polyester , wool, cotton
Recycled Threading
Recycled sweater mittens made from old preloved wool and wool blend sweaters, polar fleece lined with decorative button accents as well as a variety of knit and crochet accessories. we make all items in a child to adult sizes, our mitten patterns are our own design.
Sewing Knitwork
Hand sewn and knitted childrens clothes and baby sets, pillows, aprons and dolls clothes.
Casa Relief
DreamyZ Loungewear
Widows and orphans in Rwanda help to create these pyjamas and sleep shorts.Visit their website at dreamyz.ca
Soul Woven
Soul Woven is a fair trade fashion collection curating traditional Mayan creativity into innovative, modern design. Our collection consists of ethically sourced, beautiful authentic handmade pieces including boots, clothing, yoga accessories, exquisite jewelry and more. Soul Woven empowers artisans to gain independence and self-sustainability while allowing our consumers to make conscious and informed purchases. We aim to make ethically sourced products as accessible and convenient to consumers, as any product from the fast fashion industry.
Talis offers ethically sourced artisan silks and silver jewellery. We work directly with tribal communities in Cambodia and Thailand. Our core business practices are focused on economic empowerment for indigenous peoples. www.talis.ca
Unikati and Co.
Unikati & co is a carefully edited collection of ethically handcrafted goods from around the world. Dina and Leila, the founders of unikati & co, are committed to bringing you beautiful pieces, the creation and sale of which contributes to the empowerment of fairly-paid artisans and their families. Visit their website at unikati.ca
Everything Chocolate
Chocolate suckers for all occasions…chocolate dipped marshmallow . . . chocolate dipped pretzel rods… chocolate pizza's . . . clusters . . . bark . . . chocolate dipped rice crispy. Everyone is individually gift wrapped and homemade.
Firebat Coffeee
Firebat Coffee is etically sourced and sustainably grown. We work directly with a group of coffee makers from ElSalvador and through this direct trade they get significantly higher pay than Fair Trade. All our coffee comes from high altitude, shade grown coffee farms, where coffee is grown in harmony with nature.
Happy Bees Apiary
Happy Bees honey provides 100% Ontario honey and is a member of the On Bee Keepers Association. All of our honey is unpasteurized and we create unique flavored honey like cherry, lemon, cinnamon and more . www.happybeesapiary.ca
JoJo's Toffee
Delicious chocolate nut toffee.
Missies Streudel
Freshly baked apple, blueberry and cherry streudel.
Moose River Preserves
Fine gourmet preserves.
My Tea Brew
Selection of superb loose leaf tea blends by certified Tea Sommelier.
Nonnas Cannoli
Enjoy the creamy goodness of the sweetened ricotta filling. These Cannoli Tradizionale will not dissappoint.
Pops by Janice
Delicious holiday decorated cake pops in white chocolate or red velvet cake.
Soup Girl
Soup Girl's artisanal vegan dry soup mixes are healthy delicious and so simple, each jar makes 8 cups of soup, just add water, cook for an hour. A range of flavors available, including spicy coconut lentil, smoky split pea, Italian rice & lentil and carribean rice & peas.
Sweet and savoury shortbreads
Chocolate covered shortbread, toffee shortbread, coffee shortbread and parmesan rosemary shortbread.
Tease Cuisine
Cuisine is passionate about food and specializes in everything charcuturie. Some of our items include chutney, infused oils, cured meats, garlic confit, seasoning salts and dry rubs, pickled vegetables and more. www.teasecuisine.com .
Truly Baked Gourmet Granola
Delicious gourmet granola handcrafted in small batches by a holistic nutritionist. 5 varieties to choose from including pumpkin spice, maple, chai and cinnamon walnut. All free of refined sugar, dairy, wheat or preservatives.
Glass Butterfly
Fused glass artist, bowls, dishes, coasters, etc., unique but functional glass art.
Stained Glass by Brenda
Himelli Creations
Handmade tradional Finnish ornaments.
Intoxicative Upcycling
My creativity continues into the art of candle making, combined with the ingenuity of upcycling. I give old bottles new life as unique, clean burning soy candles. All of my artist creations are intended to make your home a nurturing sanctuary for mind, body and soul. www.etsy.com/shop/intoxicative .
Jan Jang Originals
Jan Jang creates very unique gifts including cork hotplates, hand painted garden prayer flags and personalized room signs.
Little By Little
Hiring women at risk of relinquishing their rights due to poverty in Haiti, selling their Christmas ornaments, metal art and hand crafted jewellery. Keeping families together, also orphan prevention through job creation
Sew Happy Together
Machine embroidered gifts and treasures for everyone
Belinda Carmichael Jewellery
Beautiful, unique handmade sterling Silver Jewellery and Gifts. Custom orders welcome. Visit their website at etsy.com/shop/BelindaCarmichaelSJ
Bella B You Jewels
Bella B You handcrafts one of a kind jewellery pieces. I create wearable everyday pieces using semiprecious gemstones, classic pieces to wear with that little black dress and romantic pieces to wear where I incorporate vintage beads and embellishments. Like us on Facebook at Bell B You
Designs By Craig Ewing
I design and create one of a kind wearable art and use Japanese beads, fresh water pearls and swarovski crystals in my earrings and my metal. Everything is nickel and zinc free.
Dragonfly Girl Jewellery
At Dragonfly Girls jewellery, we create handmade jewellery for special girls and women, including necklaces, bracelets, earrings, rings and pendants, beaded bookmark and keychains, chain maille and beaded wirework creations. We also do birthday parties, private lessons and jewellery repair. Find out more at www.dragonfly.ca
Earth Candy
Jewellery and hand knitted toques.
Glitterfly Designs
Handcrafted artisan jewellery made in Missisauga
Linx Jewellery
Unique jewelry designed and handcrafted by Sally Taylor, semi-precious stones, Swarovski, shells, wood and glass, etc. Visit their website at linxdesignz.com
Small World Jewellery
Quality handmade jeweler from Africa that supports several organizations that empower Africans to operate businesses and offer employment at strong wages under fair trade principals. Visit their website at smallworldjewellery.ca
Strung Out On Pearls
Unique designs meticulously created for confident, passionate, adventurous vibrantly unforgettable you! Jewellery that gets noticed.
Knitz 'n Bitz
An assortment of hand knitted items for children and adults (cowl & infinity scarves, unique hats, dishcloths, animals, decorative snowmen) as well as fleece and flannel blankets for children.
Sewing Knitwork
Hand sewn and knitted childrens clothes, toys, baby sets, pillows aprons and dolls clothes. Great for Christmas gifts.
Earn n' Learn
works with slum children and teenagers, they are given healthy snacks, tutoring, mentorship and an opportunity to provide income for their family in a safe environment making paper crafts. On the back of each card is the name of the child who crafted it as a testimony to the change you have made in their life. For more information go to www.earnlearn.org
Morning Flower Designs
Unique and creative designs featuring a combination of pressed local wild flowers, coloured paper and water coloured paints. Designs range from cute to whimsical to sentimental and are available as originals as well as prints. www.etsy.com/ca/hop/morningflowercrafts www.facebook.com/morningflowerdesigns
Carolyn Showler Pottery
Handmade stoneware pottery with original designs and glazes.
Functional Pottery
In this modern fast paced world, where everything is massed produced, I love the idea of making functional pottery that has some soul. I try to incorporate in my work that the piece has been made by hand. I intentionally leave my finger prints or allow my throwing lines to be seen, also I enjoy adding hand carved detail to give texture and to remind the user that this is a piece of pottery made by loving hands.
Hand-painted ceramic serving platters and dishes by local artist Gillian Sych.
Agnes Art Creation
Handmade Angel Collection, Christmas ornaments and handmade leather collection.
Sav's Christmas Ornaments
Timeless Jewellery Designs
Timeless Designs features unique Christmas and Holiday ornaments made of glass or polymer clay. Each ornament is designed and handcrafted by Barbra or Richard Darwin.
Decorate Your Own Snowman
For kids or kids at heart. Each kit includes a decorative hat, eyes, carrot, nose, mouth, scarf, buttons, beautifully co-ordinated and presented in a reusable kit. Great for children, family & hostess gifts.
BWA Woodturning
Functional Wood – turnings – coffee scoops, backscratchers, magnifiers, nutcrackers, kaleidoscopes, stylus, letter openers, peppermints. Visit their website at bwawoodturning.com
Philthy Woodworks
Handcrafted, original, rough, natural grain wood products and custom builds by a local Lakeshore woodworker.
The Rustic Path
At the rustic path we love wood. New, old, recycled and salvaged, we use it all to create unique house and holiday decor and wooden signs for you home.
Wine Crate Crafts
Hand crafted wood trays and containers made from high end French, Italian & Spanish vintage wine crates. The trays can be used to serve or as gift baskets filled with your gourmet items etc.
Beagle Bakery
Natural Grain free cakes, cookies and so much more for your Dog. Personalized People & Pet Christmas Ornaments. www.beaglebakery.com
Cat Fancies and Cuddly Critters
Luxuriou cozy snugglebeds, pet mats and kitty caves. Handcrafted premier quality catnip toys made with organic catnip. Only the best will do for your favorite friend.
David Cook
Local historian and author of Fading History Vol. 1 & 2.
Eco Girl
Eco friendly products made from bike inner tubes and other materials. Visit their website at ecogirl.ca
Nicoles Doll Clothes
Bookmarks are handcrafted with beads and new or upcycled charms.

subset: commoncrawl


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By Michael S. Goldberger, film critic
Seth MacFarlane's "Ted" is a kid's fairy tale strictly for adults. And although you get the feeling the filmmaker is breaking new ground, one isn't quite sure in what way that is. Clearly, however, this is a sweet, nutty and rather raunchy romp through the imagination, deliriously blurring the line between child fantasy and so-called grownup responsibilities.
Once upon a time, about 20 or so years ago, there was a shy little boy named John Bennett who had no friends, and little prospect of ever accumulating any. Thus, one Christmas, to ameliorate the situation, his parents bought him a Teddy bear. They became fast friends. Only problem was, Teddy wasn't real. So little Johnny made a real big wish.
Fast forward to the present and the once heralded Christmas miracle is all but forgotten by a fickle media that made it the cause celebre back when the title character first took human breaths. But that's OK with John and Ted, who've been as inseparable as peas and carrots lo these many years. And that's OK with John's girlfriend, Lori…or so it seems.
But dig a little deeper and you'll mine the main plot in this oddly shaped love story. While Mila Kunis's Lori, a career lady who shares digs with the duo, appreciates the kinship she has ostensibly turned into a triangle, she worries that her beau is limiting his horizons, working in a dead-end car rental job by day, getting stoned with Ted by night.
Fact is, John is aware of her concern, fully acknowledging his curious case of arrested development and the downside of maintaining so devoted a link to such a fantastical childhood. Yet he feels indebted. What would Ted do without him? But most of all he's having a great time, stuck in perennial adolescence with his bear. What to do, what to do?
Adding heady substance to auteur MacFarlane's creative scenario, Ted isn't completely selfish, but rather in sympathy with the tragic dilemma his brother under the fur faces. That's the wonderfully strange thing. He's real. If we cut him, doth not his stuffing escape? Great F/X and Mr. MacFarlane's emotive voicing make him fully alive to us.
Mr. Wahlberg, as the would-be adult Johnny Bennett, is quite good in a difficult role, responding to and engaging a cinematic chimera, a cutting edge variation on Jimmy Stewart's tête-à-tête with Harvey. But the question this fictional relationship ultimately begs is, can an animated character win an Academy Award? You can make a case for it.
Of course, it's always possible the Academy will shun the ursine phenomenon because of his foul-mouthed ways. Plus, he's not part of the Hollywood establishment. But aside from the incredible fact that he exists only on film, it'll take some doing to top his totally enrapturing portrayal. Yep, his persona even matures with the circumstances, so to speak.
Simple of plot, the screenplay assigns judge, jury and prognosticative duties to the viewer, who is beset with figuring out how this sticky wicket of a social dilemma might be resolved. While we'd like to see Lori and Johnny live happily ever after, we can't entirely fault party animal Ted for whatever encumbrances to that goal he represents.
After all, he didn't ask to be wished into existence. And he's known little else but to make his best pal happy. Similar to twins who've developed their very own language, for John and Ted moving on with their lives means regaling each other with one memorable anecdote after the next, needing only a buzz word to incite a mutually enjoyed cackle.
Here, Mr. MacFarlane shows us via a cleverly eccentric monograph within his movie that he knows true male friendship, or bromance as film writers have come to call it of late. Ted and Johnny's residence in a constant private joke is a testament to their bond and a declaration of dedication to a gesellschaft, whether they know what it means or not.
And maybe no one understands that fact better than Lori, impressively realized by Miss Kunis. Secure in her sense of self worth, figuring there's enough love to go around for all those concerned, Lori is confident in Johnny's love. But there'd be no dramatic purpose here if that confidence weren't intriguingly shaken by a series of hilarious complications.
Consistently spiced with Ted's ever-abashing, street corner profanities, the Boston Southie dialogue is integral to the comedic lilt as the film mind-snatches you into its screwy world. Complemented by allusions to pop culture, politics and anything else that strikes MacFarlane funny, "Ted" is a bizarrely delightful bearer of movie entertainment.
"Ted," rated R, is a Universal Pictures release directed by Seth MacFarlane and stars Mark Wahlberg, Mila Kunis and the voice of Seth MacFarlane. Running time: 106 minutes
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Thursday, August 9, 2012
It went from being a quilt I was uberexcited about to a quilting nightmare in one fell swoop.
I knew it was going to be hard because I was using the tinier Janome to quilt it.
I have to take Foxy in to get checked for the grease issue.
I started quilting it....and the petals that I had spent HOURS ironing on started coming off.
I don't know if it was the humidity or the pulling on the quilt top, but it was ridiculous.
And maddening. I wasn't going for an unfinished edge applique, but I'm not confident it's not going to fray like mad. I am going to try to steam it again before I wash it, but BLARG.
I don't think I'm ever going to use Pellon Wonderunder again. It was odd. I've used Pellon products before and never had trouble before, but this was redonkulous.
Anyway, here it is, prewash:
Please ignore any threads.
I also caught up on some bee blocks:
Neutral and Not
Running with Scissors
I still have to do Nubees, catch up on the Sew Intertwined QAL and How Far will you go QAL, and sew the binding on the Across the Sea boy's quilt....and start handquilting the Medallion.
Yarg. I spent hours today organizing kid folders. I don't know if I have to go into work tomorrow or not. Right now, the dog is all fretty from a thunderstorm.
8 comments:
i love how you used uberexcited, blarg, and redonkulous all in the same post! you are super awesome! ilove your blocks and the quilt looks wonderful even though you wanted to kill it!ReplyDelete
Hi Ella - I also gave up on the Pellon fusibles. They never seemed to stick well (or at all!). I do like the Heat n Bond products, though, and haven't had any of the same problems with either the regular, lite, or ultra product of theirs.ReplyDelete
The quilt looks fantastic, as do the blocks - it'll be OK I'm sure - I would say set the iron up to 'scorch', but I don't want to be responsible for the result!ReplyDelete
Oh no! So sad to hear of so many hours of ironing turned yucky. I'm ironing my petals right now and hoping desperately that it works. I think it turned out looking really lovely, I'm sure you're glad to be done with it!!ReplyDelete
The blocks look great :)
Bugger on the wonder-under. I've had things like this happen occasionally with bondaweb, but never worked out why it happens sometimes and not others o.OReplyDelete
What a great job on the quilt! I've used some really bad fusible web before too. Loving the granny blocks! =DReplyDelete
Boy your last 2 quilts have just had some rough spots - first grease and then bad fusible, that just stinks!!! The quilt looks amazing regardless of how much you struggled, it really does look FANTASTIC!!! I love all the rest of your blocks too :)ReplyDelete
It still looks wonderful, at least the in the picture. So sorry that the fusible wasn't playing nice.ReplyDelete


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Early works were published by Experimental Music Catalogue. This stopped trading in 1981, but is now run by Christopher Hobbs and Virginia Anderson. For information on these works contact email@example.com
All Gavin Bryars' music is now published by Schott Music, 48 Great Marlborough Street, London W1F 7BB
Works not published by Schott or EMC are marked with an asterisk (*) and are unavailable for performance.
NB. All Gavin Bryars' manuscripts have been acquired by the British Library. These may be accessed through Dr. Nicolas Bell, Curator Music Collections, The British Library, 96 Euston Road, London NW1 2DB
There are a number of works, early attempts at composition and so on, some of which were performed. List in progress…
Three no. 2 "Visions" (Mss. In Notations Collection of John Cage)
Hoyu spoke no words (Mss. In Notations Collection of John Cage)
Three no. 7
16 Fragments for solo guitar
Instrumentation: Any number of keyboards, including one prepared piano.
Duration: indeterminate
Published in EMC Keyboard Anthology.
First performance: Kingston College of Art, 13 December 1968.
Mr. Sunshine (1968)
Mr. Sunshine is one of three pieces written in 1968 for the pianist John Tilbury following my return from America. It consists of one large page of notation incorporating certain elements of indeterminacy. There are "looped" areas where short phrases are played over and over, and some of these are connected by lines to other parts of the page, giving the possibility of moving through the music in a series of leaps. Although written for a solo pianist, it is strictly for any number of pianos, of which one should be "prepared". This prepared piano may be used to maintain a pulse, to colour other material or to give a continuum to the freer, non prepared part.
It is, in fact, the earliest piece that I still acknowledge, all others having been destroyed.
Solo theatre piece(uses special garment).
Duration: indeterminate.
Published in EMC Visual Anthology.
First Performance: University College, Cardiff, 6 November 1969.
Tape piece (1/2 Track Stereo)
Duration: c.30 minutes
First performance: Systems Art exhibition, Helsinki, 1970
Indeterminate(text notation)
Published in EMC Verbal Anthology.
First performance: Reardon-Smith Hall, Cardiff, 1969
Instrumentation: Indeterminate (possible materials include stereo tapes, string ensemble, percussion, low brass, brass quartet, bass clarinet, cassette tapes of speech, keyboard, 35 mm slides, visible sound effects, music box).
Duration: versions of 25', 35', or 1 hour (plus)
Published in Soundings 9 (USA).First performance: Queen Elizabeth Hall, London 1972
The Sinking of the Titanic (1969- )
This piece originated in a sketch written for an exhibition in support of beleaguered art students at Portsmouth in 1969. Working as I was in an art college environment I was interested to see what might be the musical equivalent of a work of conceptual art. It was not until 1972 that I made a performing version of the piece for part of an evening of my work at the Queen Elizabeth Hall, London and during the next three years I performed the piece several times. In 1975 I made a recorded version for the first of the ten records produced for Brian Eno's Obscure label. In 1990 I re-recorded the piece 'live' at the Printemps de Bourges festival when the availability of an extraordinary space - the town's disused water tower dating from the Napoleonic period - and the rediscovery of the wreck by Dr. Ballard made me think again about the music. In any case the piece has always been an open one, being based on data about the disaster but taking account of any new information that came to hand after the initial writing. This version forms the basis for the 1994 recording on Point.
All the materials used in the piece are derived from research and speculations about the sinking of the "unsinkable" luxury liner. On April 14th 1912 the Titanic struck an iceberg at 11.40 PM in the North Atlantic and sank at 2.20 AM on April 15th. Of the 2201 people on board only 711 were to reach New York. The initial starting point for the piece was the reported fact of the band having played a hymn tune in the final moments of the ship's sinking. A number of other features of the disaster which generate musical or sounding performance material, or which 'take the mind to other regions', are also included. The final hymn played during those last 5 minutes of the ship's life was identified in an account by Harold Bride, the junior wireless operator
"...from aft came the tunes of the band.....The ship was gradually turning on her nose - just like a duck that goes down for a dive... The band was still playing. I guess all of the band went down. They were playing "Autumn" then. I swam with all my might. I suppose I was 150 feet away when the Titanic, on her nose, with her afterquarter sticking straight up in the air, began to settle slowly.... The way the band kept playing was a noble thing... the last I saw of the band, when I was floating out in the sea with my lifebelt on, it was still on deck playing "Autumn". How they ever did it I cannot imagine."
This Episcopal hymn, then, becomes the principle element of the music and is subject to a variety of treatments and it forms a base over which other material is superimposed. Although I conceived the piece many years ago I continue to enjoy finding new ways of looking at the material in it and welcome opportunities to look at it afresh.
Unspecified ensemble.
Duration: indeterminate
Published in EMC Rhythmic Anthology.
First performance: BBC 2 'Art and Technology' series, November 1970
Recording piece
Published in EMC Verbal Anthology.
Never performed - imperfectly realised Waterloo Station, summer 1970.
Instrumentation: speaking voices, tape
Duration: indeterminate
Published in EMC Verbal Anthology.
First performance: Portsmouth College of Art, 14 January 1970.
Theatre piece for 2 singers.
Published in EMC Visual Anthology.
First performance: Black Swan, York, 30 November 1984
Instrumentation: 1 or 2 prepared pianos.
Duration: c.25 minutes
Published in EMC Keyboard Anthology.
First performance: Purcell Room, London, 9 October 197O
Instrumentation: Electronics
Published in EMC Visual Anthology.
Imperfect private performance, Portsmouth College of Art, November 1970
Environmental piece.
Published in EMC Visual Anthology
Never performed.
Unspecified ensemble, dedicated to John White.
Duration: c.20 minutes
Published in EMC Verbal Anthology.
First performance: Bluecoat Hall, Liverpool.
Instrumentation: 1 player, 2 guitars (or multiples of this)
Published in EMC Rhythmic Anthology
First performance: Studio recording Incus Records (Derek Bailey guitars).
First live performance: Queen Elizabeth Hall, London, December 1972 (Derek Bailey/ John Tilbury, 2 players 4 guitars).
Instrumentation: Pre-recorded Tape and ensemble
Duration: 25' 40' 60' or 74'
First performance: Queen Elizabeth Hall, December 1972.
Jesus Blood Never Failed Me Yet
In 1971, when I lived in London, I was working with a friend, Alan Power, on a film about people living rough in the area around Elephant and Castle and Waterloo Station. In the course of being filmed, some people broke into drunken song - sometimes bits of opera, sometimes sentimental ballads - and one, who in fact did not drink, sang a religious song "Jesus' Blood Never Failed Me Yet". This was not ultimately used in the film and I was given all the unused sections of tape, including this one.
When I played it at home, I found that his singing was in tune with my piano, and I improvised a simple accompaniment. I noticed, too, that the first section of the song - 13 bars in length - formed an effective loop which repeated in a slightly unpredictable way. I took the tape loop to Leicester, where I was working in the Fine Art Department, and copied the loop onto a continuous reel of tape, thinking about perhaps adding an orchestrated accompaniment to this. The door of the recording room opened on to one of the large painting studios and I left the tape copying, with the door open, while I went to have a cup of coffee. When I came back I found the normally lively room unnaturally subdued. People were moving about much more slowly than usual and a few were sitting alone, quietly weeping.
I was puzzled until I realised that the tape was still playing and that they had been overcome by the old man's singing. This convinced me of the emotional power of the music and of the possibilities offered by adding a simple, though gradually evolving, orchestral accompaniment that respected the tramp's nobility and simple faith. Although he died before he could hear what I had done with his singing, the piece remains as an eloquent, but understated testimony to his spirit and optimism.
The piece was originally recorded on Brian Eno's Obscure label in 1975 and a substantially revised and extended version for Point Records in 1993. The version that is played by my ensemble was specially created in 1993 to coincide with this last recording.
Instrumentation: indeterminate
Published in EMC Verbal Anthology.
Never performed.
Instrumentation: Indeterminate
Published in EMC Visual Anthology
Never performed.
Environmental piece
Published in EMC Visual Anthology.
Incomplete performance only.
Duration: 10'
Instrumentation: Up to 3 strings and piano.
Published in EMC String Supplement.
First performance: University of Keele, 16 November 1983.


text:
Our Mascot
When the company was formed in 2007, the face of Poseidon the founder's Cat was used as the company logo. In 2013 the logo was refined to the Trident with rings that is used today. The Cat is now the company's mascot and never far from our memories.
Our Cat
Our Logo
Our Mascot
Why Poseidon? Why the cat? Why POSH?
Poseidon was the Greek god of the sea and controlled the safety and destiny of mariners. Where old fashioned sailors trusted to a god, modern sailors trust in themselves, and POSH people are there to support them.
When naming the company we wanted a title that would still mean something when "initialised" So it is no coincidence that the name breaks down to provide a shortened version of the name; POSH.
In itself POSH is reputed to be a marine phrase developed from "Port Out, Starboard Home". This designation on ones luggage guaranteed a cabin on the North side of the ship to and from India, thus it would be cooler and more comfortable. Of course this choice was more expensive and exclusive and the abbreviation soon stuck to the social strata that could afford this. Some correspondents have pointed out that it might be an urban myth, but still it is a nice story whether it is true or not.
Poseidon was also the name of one the founder's cats and it is her face that is depicted in the company logo. Sadly the eponymous cat died in 2006.
In researching the advisability of using a cat's head as the company logo the following information came to light;
"Mi-Ke, or tri-coloured cats, (similar to Poseidon) were taken by Japanese sailors on their ships to bring them good luck." The various colours of the cat apparently brought protection and security from various woes: pestilence, storms, and ill fortune so that voyages would be safe and profitable. The more colours the cat carried the luckier they would be.
" Sailors believe that if a ship's cat mews and appears to be cross, they will face a hard voyage – but if it is bright and lively, there will be a brisk "following" wind. It used to be said that a contrary wind at sea could be raised by shutting a cat in a canister. Throwing a cat overboard resulted in an immediate storm. No sailor would dream of doing this, however, since it was considered good luck to have a cat on board."
The association with cats and the sea has always been a long and respectful one with cats conferring good luck upon vessels that carried them, quite apart from keeping down the rats and mice.
The position formerly occupied by the ship's cat, that of protecting the vessel and bringing "luck" or good fortune, is now where the Safety Management or ISM system sits. We no longer rely on luck or the reputed effects of a feline presence but on structured management and the control of the risks and hazards of our adventures.
Poseidon Offshore (Safety and Health) Ltd is a company that can be called upon to assist, support, audit and inspect these features in the modern idiom, but it also carries with it the generations old veneration of the cat as man`s protector at sea.


text:
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Stockpile List for Patch 5.2: Make Thousands on Patch Release! All Professions & Auction House Flippers
Stockpile List for Patch 5.2: Make Thousands on Patch Release! All Professions & Auction House Flippers
What's up guy and girl players of WoW. Tarou here and I'm bringing you a video telling you what items you need to stock up on for Patch 5.2.
This patch could come as early as February 26th and it's bringing a lot of new items, dailys, raiding content, and a new PvP season. That means tons of players will be getting new gear and they're going to need enchants, gems, leg armors/threads, and buckles.
They'll also need flasks, potions, and other things. For the fresh 90s or players just entering raiding or pvp'ing on their alts or main, they'll need everything including gear. This patch introduces new craftable gear for PvP covering all slots aside from weapons as of right now. I don't know if they'll add craftable weapons but even if they do, most the mats I list will cover them anyway.
Dump List:
All Contender's gear within a month of release. (Players won't know immediately there is better gear and some will still buy the older Contender's PvP gear.)
Farming/AH reselling:
All mats for max level listed under the profession it's used for.
Leg Enchants
Windwool Cloth
Windwool Bolts
Don't forget your CD daily
Leg Armor
Mining & Herb Bags (Don't go crazy.)
Magnificent Hides
Exotic Leather
All max level enchant mats.
All popular max tier enchants.
Don't forget your CD daily
Ghost Iron Bars
Living Steel
Living Steel Belt Buckles
Ghost Iron Bars
Living Steel
All max level popular gems/metas
Uncut max level Gems/Primal Diamonds
Serpent's Eye
Ghost Iron Ore
Don't forget your CD daily
Max level Flasks (Elixir spec only)
Max level Potions (Potion spec only)
Living Steel (Use your CD everyday!)
Trillium Bars (Trans)
Ghost Iron Bars
Wild Jade, Vermilion Onyx, Imperial Amethyst (Trans)
Golden Lotus (Lots - Elixir spec)
Don't forget your CD daily
Max level herbs (Least expensive for milling)
Epic Shoulder Enchants
Don't forget your CD daily
Ghost Iron Ore/Bars (Go Crazy!)
Trillium Ore/Bars
Max level Herbs
Golden Lotus (Lots)
Magnificent Hides
All max level leather especially Exotic
I may have missed a few items so I'll update this list as need be on my website at tarouwowguides.com. Also keep in mind that some things may be speculative and Blizzard can change things with the live patch. There's always a risk but I eliminate it as much as possible.
Finally, this is a good time to clean up and clean out your bank, bags, and alts. You're going to need the space and having everything organized will help making gold a lot easier with less chaotic frustration.
You also want to make sure you have all the best money making addons like the ones listed on the screen. Each has a video tutorial if you need help setting them up.
Well, that's it and I wish the best of luck to everyone in your gold making adventures in Patch 5.2.
Thanks for watching and if you haven't yet please subscribe to my YouTube channel, http://www.youtube.com/ryutaroakatarou and share this video!
"Now Go Stock Up Your Way to Riches!" Lates.


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Not another post. I know I should slow down with my updates, but when it comes to self-promotion I'm simply insatiable.
So what have I been up to since last we spoke? Well, I pretty much took a year off stand-up to focus on getting my directing career going. Pretty successful – did the EIFF talent lab, directed my first ad, and now I have 4 feature scripts ready to go, so I'm aiming to shoot something this year. Watch this space – I'll no doubt let you know 6 months after it's been released.
In the meantime, I'm back on stage again, so you can come and see me live if you've got nothing better to do, which you clearly haven't because it'll be excellent. The dates are on the live section tab now.
God, I just realized I didn't even post here when The Boy with a Camera for a Face went up on Vimeo and got staff picked. What is wrong with me? I should probably put up a link to that too. Here it is: https://vimeo.com/151493973
Well, that's it for another few weeks/months/years. You should really follow me on twitter – I'm better at that – there's usually a bi-monthly tweet.


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The Living List28 November, 2014
Beagle Street is pleased to announce the launch of The Living List – a new online hub where you can create a list of the things you want to do, to be happier and help you to enjoy life to the max.
At Beagle Street, we believe that celebrating your passions is key to being happy. To get you started, we've teamed up with experts in different fields to create a variety of different Living Lists, which include time travelling with the beautiful Lil Bub to playing on a black and white diamond encrusted chess board with House of Coco.
Are you a foodie with a passion for eating out, or an ambitious individual with a focus on your career? Use the existing lists for inspiration and select the experiences that most appeal to you to create your very own Living List – based on your own interests.
The Living List Competition
To celebrate the launch, we are giving one lucky winner the chance win up to £1500 towards making what tops their living list a reality. Entries can be as creative or simple as you like – just as long as it reflects your persona. To find out more, enter the competition* or to view entries click here:
From big aspirations to day-to-day activities, be inspired by the Living List, and start living your list today.
So go on and start your Living List here:
Live it. List it!
*Terms and Conditions apply. Open to people who are 18 years and above. Closing date 31 January 2015.


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Multifamily Energy Savings Program (Existing Buildings and New Construction) (Wisconsin)
Last modified on February 12, 2015.
Financial Incentive Program
| Wisconsin
|Multifamily Energy Savings Program (Existing Buildings and New Construction)
|Incentive Type
|State Rebate Program
|Applicable Sector
|Multi-Family Residential
|Eligible Technologies
|Boilers, Building Insulation, Central Air conditioners, Chillers, Clothes Washers, Dehumidifiers, Dishwasher, Duct/Air sealing, Energy Mgmt. Systems/Building Controls, Equipment Insulation, Furnaces, Heat pumps, Heat recovery, Lighting, Lighting Controls/Sensors, Motor VFDs, Refrigerators, Water Heaters, Geothermal Heat Pumps, LED Lighting, Tankless Water Heaters, Parking Garage Exhaust Controls
|Active Incentive
|Implementing Sector
|Energy Category
|Energy Efficiency Incentive Programs, Renewable Energy Incentive Programs
| Prescriptive
Varies by equipment type
|Equipment Requirements
| Varies by equipment type; equipment must be new to qualify for incentives
Geothermal Heat Pumps must be variably controlled
|Expiration Date
|2013-12-31 (most recent applications)
|Funding Source
| Focus on Energy
|Maximum Incentive
| Prescriptive: Varies by equipment type, $200,000 per project
Custom: Lesser of 50% of cost or $200,000 per project
| http://www.focusonenergy.com/business/efficient-facilities/multifamily-energy-savings
Focus on Energy offers incentives to owners of existing apartment and condo buildings who make energy efficiency improvements. The program offers a combination of technical assistance, prescriptive equipment rebates, and custom incentives based on the amount of energy that a project saves. Eligible equipment generally includes energy efficient appliances, HVAC equipment, building shell improvements, and lighting measures. Incentives are available for common area and building improvements, as well as certain improvements made in individual dwelling units.
To begin participating in the program, eligible building owners/managers work with a Focus Energy Advisor to perform a free energy assessment and identify suitable building improvements. For prescriptive HVAC and appliance rebates, up to $10,000 in incentives are available without pre-approval. Custom incentives for electricity and natural gas savings are broken down into three tiers based on the estimated energy savings. View the program web site for a full list of prescriptive and custom incentives for existing multi-family buildings. The Focus on Energy program itself does not have a specific expiration date, although many individual program applications expire at different times throughout the year.
Focus on Energy works with eligible Wisconsin residents and businesses to install cost effective energy efficiency and renewable energy projects. Focus information, resources and financial incentives help to implement projects that otherwise would not be completed, or to complete projects sooner than scheduled. Its efforts help Wisconsin residents and businesses manage rising energy costs, promote in-state economic development, protect our environment and control the state's growing demand for electricity and natural gas.
Please note that customer eligibility for Focus on Energy incentives is based on utility participation; interested individuals and companies should use the eligibility tool on the program web site to determine if their local utility participates in the Focus on Energy program. Further incentive details, applications, and other information are available at the program website.
Incentives are also available for multifamily new construction. Owners and landlords of existing rental buildings in the Wisconsin Public Service territory may also be eligible for additional incentives and technical assistance. Interested customers should contact Focus on Energy.
Incentive Contact
|Contact Name
| Focus on Energy MESP
|12075 Corporate Parkway, Ste. 100
|Mequon, Wisconsin
|Zip/Postal Code
| (800) 762-7077
- Incentive and policy data are reviewed and approved by the N.C. Solar Center's DSIRE project staff.[1]


text:
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Be the first NJ.com user to add photos or videos of Morin Marla
Marla Morin is located at 500 Barnett Pl in Ho Ho Kus and has been in the business of Educational Consultant since 1984.


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We're still half a year away from the 85th annual Academy Awards, but it turns out the host and producer were almost already decided.
Before he left his position as Academy of Motion Picture Arts and Sciences president, Tom Sherak reportedly tried to lock down Jimmy Fallon as host and Lorne Michaels as producer for next year's Oscars. The Los Angeles Times and Deadline offer up two different versions of the events, and it's unclear whether Fallon is still a contender for the position now that Sherak is gone.
Apparently one of the big issues with having Fallon as host is the fact that he has long been a staple of NBC, the rival network of ABC. Both "Saturday Night Live" and "Late Night with Jimmy Fallon" are NBC shows, and the latter is a direct competitor to ABC's "Jimmy Kimmel Live!" So despite Fallon's good run at the 2010 Emmys and his hosting gigs at the 2009 VMAs and 2006 and 2001 MTV Movie Awards, it sounds like this actually may not be his year to host the Oscars after all.
While I think Fallon is a funny comedian and enjoy his late night show, I don't think he would make for the best Oscars host. He's a goofy, giggly guy, which certainly has its place in the comedy world, but doesn't quite fit with the self-important Academy Awards persona. I can just picture him breaking out laughing in the middle of a sketch that he's supposed to be performing and cringe.
At the same time, Fallon has really blossomed in his years as a late night TV host. He's got a great relationship with many celebrities in Hollywood and always manages to get some great material from them. I'm sure he could pull off some great bits during the show, but that might not be enough to support the entire event. Honestly I think he needs a few more years under his belt: look at Anne Hathaway and James Franco's hosting gig and compare it to last year's performance by Billy Crystal.
It does seem smart to bring a comedian on as host, though. And late night celebs like Jon Stewart and David Letterman have done a great job at the Oscars in the past. I think that Jimmy Kimmel would actually make for a great host this year, which would work out better for ABC. He also has great relationships with celebs (please go watch "I'm F--king Ben Affleck" right this second if you haven't already) and his humor can be both grounded and goofy at the same time (look at his "Celebrities Read Mean Tweets" video), all of which are elements the Academy is likely looking for. He's an everyman, and that's a quality that works great in these circumstances. Fallon would probably do a fine job as a host too, but I personally wouldn't be too heartbroken if it took him a few more years to get the gig.
Do you think Fallon would be a good host for the Oscars? Tell us in the comments section below or on Twitter!


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Open Thread: Talk About What Matters Most to You
Discuss and debate the issues that mean the most to you.
So, North Carolina passed a law, HB2, that, among other things, declares that people should use the bathrooms that match the "sex" on their birth certificates, regardless of their gender. It's a terrible, no-good, very bad bill.
Meanwhile in Mississippi, the state legislature passed a sweeping "religious liberty" bill that basically allows conservative Christians to do whatever they want and then claim they're protected by nature of their faith. These religious freedom bills are currently trending in conservative circles, and they are bad news bears.
Let's be clear here: Such bills are unacceptable, and the lawmakers who voted yes on them, and the governors who signed off on them, are bigots.
Let's also be clear about something else: North Carolina and Mississippi, as entities, are not bigoted. They're filled with a wide variety of people, some of whom are bigots who vote other bigots into office, some of whom are the very groups affected by the laws those bigots pass, some of whom are passionate advocates, some of whom are neutral and confused, and want to learn more.
This is why I am infuriated by repeated calls to boycott both these states from people who claim that this is some kind of grand gesture that will really show those bigots, yes siree. This is patently wrong, and it's also extremely harmful to the very populations that these brave protesters, led by irritating groups like the HRC, claim to be helping.
If you want to help a population that is being oppressed by its government, totally withdrawing all support and connections with the outside world is not the way to do it: Imagine if we saw Wendy Davis filibustering in an anti-abortion Texas and promptly pulled all our support out of Texas on the grounds that the legislature was behaving unconscionably. Instead, when people descended upon the state house to make a stand, we sent supplies, cheered them on from afar, traveled in from out of state, and spoke out against oppression by buying in to an effort to fight it.
Here's the thing about boycotts that many people don't seem to understand: When used effectively, they can be a powerful tool, but when applied indiscriminately, they can actually be extremely harmful.
In the case of these two great states, here's who's not being hurt: The governors. The legislature. The corporations that don't really care about oppressive tactics — in fact, let's take a moment to appreciate the fact that the companies making bombastic claims about withdrawing from these states are still turning around and quietly making donations to the politicians who support legislation like this. BUSTED.
Here's who is hurt:
Sure, maybe you ding a few businesses owned and run by people who supported the law, and you don't give tax dollars to a legislature that passes bigoted and unconstitutional legislation. But really? That's not who's going to bear the brunt of your boycott.
Small businesses will feel it, including those who are radically inclusive and work to protect their LBGQTAI employees and customers, like, oh, independent bookstores, who are getting absolutely hammered right now.
Organizations within North Carolina that are fighting for equality and losing out because people aren't aware of their work will be the ones to suffer from the boycott, like, say, this brewery, which is cooking up a special beer to be sold across the state, proceeds going to fight this law.
The working class will take a hit, thanks to all the jobs that are being lost as a result of slowed business and corporate pullouts. Many of those people, incidentally, are not bigots, despite what stereotypes about the working class might lead you to believe, but people who have lost their jobs because of misguided boycotts who are bigots are probably going to view LGBQTAI people even more unfavorably because now they have something else to blame them for.
Oh, and, mostly importantly: LGBQTAI people.
You know what sucks? Being marginalized and isolated and alone in a landscape where everyone hates you and it feels like there is nowhere safe to go. Feeling like no one is there in solidarity with you, and being acutely aware that everyone from outside your state assumes that everyone in your state is a rampaging asshole.
You know what really sucks? Being a young person surrounded by bigoted laws and hostility who doesn't get to meet LGBQTAI musicians, authors, and other entertainers and performers who have decided to boycott your state because they think they're "making a statement."
I see a growing list of performers pulling out of shows and tours in the region, and it breaks my heart, especially when it comes to young adults who really need to see these figures in their lives, especially right now. They need defiant, in-your-face real adults who show up to say, "Look, this is survivable, and we are gonna fight it, together." They need people like Joseph Fink, who pointedly explained why Night Vale will keep its promises to North Carolina.
They need people like Laura Jane Grace and Against Me!, who will be showing up to play venues in North Carolina even though she won't be able to use the bathrooms. They don't need the parade of "allies" who are making big publicity stunts out of their refusal to enter the state.
These states need a buycott. They need you to show up for the people who are doing it right: For the businesses posting inclusive posters on their windows. For the North Carolina Democrats who walked off the floor rather than participate in a sham vote. For the LGBQTAI-owned and run businesses, companies, and organizations that can't afford to move out of state, that don't want to leave the state, that are proud of who they are. For the LGBQTAI kids who need to be reassured that we are building a better world. For the trans women who are struggling to access transition care because they can't afford it, who are unable to correct their birth certificates to satisfy the state. For the LGBQTAI adults who need to know that we are standing with them.
North Carolina and Mississippi are not this. They are not defined by this, unless we allow this to define them. Don't abandon the South, and don't treat this as a product of regressive social attitudes that are only present in Southern states, because that kind of generalisation is itself bigoted, in addition to being wrong.
Boycotting them isn't heroic. Boycotting is the equivalent of locking a cancer patient in a closet and then taking her chemo and radiation away.
Photo: GoToVan/Creative Commons


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|ORASU NOU: Satu Mare County, Transylvania
US Commission No. ROCE-0564 -
The cemetery is located in Orasu Nou, 3915, judet Satu Mare, Romania at 4750 2317, 270.6 miles NNW of Bucharest and 11 km from Negresti Oas.
The alternate name: Avasujvaros (Hungarian). Present town population is under 1,000 with no Jews.
The 1880 Jewish population by census was 128, by 1900 census was 234, and in 1930 was 252 Jewish inhabitants. In May 1944, the Jews were gathered in the ghetto of Satu Mare and on May 19, 22, 26, 29, 30, 31, and June 1 were deported to Auschwitz. The unlandmarked Orthodox, Hasidic cemetery was established in second half of the 19th century with last known burial was inter-war period.
The rural/agricultural hillside, separate but near other cemeteries, has no sign or marker. Reached by a public road, access is open to all. A fence with a non-locking gate surrounds the site. Approximate pre-WWII size is unknown. Approximate post-WWII size is 48 x 32 m. 20-100 stones are visible. 1-20 stones are not in original location. Less than 25% of the stones are toppled or broken. Location of stones removed from the cemetery is unknown. Vegetation overgrowth in the cemetery is a seasonal problem preventing access. Water drainage is good all year.
No special sections. The oldest known gravestone dates from second half of the 19th century. The 19th and 20th century marble, granite, limestone, sandstone, and concrete smoothed and inscribed and carved relief decorated common gravestones have Hebrew inscriptions. No known mass graves.
The local Jewish community owns the property used for Jewish cemetery and orchard. Adjacent properties are agricultural and local cemetery. Rarely, private Jewish or non-Jewish visitors stop at the never vandalized cemetery. Maintenance has been cleaning stones and clearing vegetation. Current care is regular unpaid caretaker. No structures. Weather erosion is a moderate threat.
Claudia Ursutiu, Pietroasa Str. no. 21, 3400 Cluj Napoca, Romania, tel. 0040-64-151073 visited the site and completed the survey in July 2000 using the following documentation:


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Re: HARTZOG; Hartsock/Hertzog/Hartsog/etc.,etc.
In reply to:
Re: HARTZOG; Hartsock/Hertzog/Hartsog/etc.,etc.
The Hartsoe/Hartsaw/Hartsough family of Ashe
and Lincoln Cos., NC are descendants of John
Philip Hertzog, also called Philip Hartsoe.
Philip was the son of Friedrich Hertzog who
immigrated to America, landing at Philadelphia on 11 Sep 1741, along with
his family, including Philip, and his sons-
in-law Plickard D. Seyler/Siler and Ulrich
Weiss/Uldrick White. The Hertzogs lived at
Upper Salford, Philadelphia County, PA where
Friedrich had a farm and grist mill.Then
they moved down the Great Wagon Road to Augusta Co., VA where Friedrich had another
mill.After he left PA he advertised his
farm and mill at Upper Salford in Sauer's
(German language) Newspaper in Germantown,
PA.About 1760 all three families, Hertzogs,
Seylers, and Weiss all moved to Rocky River
in what was then Orange County (after 1784
Chatham Co.).John Philip Hertzog married
Hannah -----.He was an activist during the
War of the Regulators and was arrested with
others and tried at Hillsborough.He was
one of the fortunate ones because he was only fined.Others were executed for their
I am currently working on a paper on the
Hertzog/Hartesoes; Weiss/Whites; and Seyler/
Silers.I will send anyone who wants one
a copy as an E Mail attachment.
All three families lived on or near Rocky
River a few miles north of Siler City in
Chatham Co., NC.Siler City was named for
a later member of the Seyler/Siler family.
More Replies:
Re: HARTZOG; Hartsock/Hertzog/Hartsog/etc.,etc.
Marsha White 5/02/10
Re: HARTZOG; Hartsock/Hertzog/Hartsog/etc.,etc.
Bonnie Partin 12/13/08
Re: HARTZOG; Hartsock/Hertzog/Hartsog/etc.,etc.
Judy Jordan 9/05/00
Re: HARTZOG; Hartsock/Hertzog/Hartsog/etc.,etc.


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text:
- Methodology article
- Open Access
Cell subset prediction for blood genomic studies
- Christopher R Bolen1,
- Mohamed Uduman1 and
- Steven H Kleinstein1, 2Email author
© Bolen et al; licensee BioMed Central Ltd. 2011
- Received: 11 March 2011
- Accepted: 24 June 2011
- Published: 24 June 2011
Genome-wide transcriptional profiling of patient blood samples offers a powerful tool to investigate underlying disease mechanisms and personalized treatment decisions. Most studies are based on analysis of total peripheral blood mononuclear cells (PBMCs), a mixed population. In this case, accuracy is inherently limited since cell subset-specific differential expression of gene signatures will be diluted by RNA from other cells. While using specific PBMC subsets for transcriptional profiling would improve our ability to extract knowledge from these data, it is rarely obvious which cell subset(s) will be the most informative.
We have developed a computational method (Subset Prediction from Enrichment Correlation, SPEC) to predict the cellular source for a pre-defined list of genes (i.e. a gene signature) using only data from total PBMCs. SPEC does not rely on the occurrence of cell subset-specific genes in the signature, but rather takes advantage of correlations with subset-specific genes across a set of samples. Validation using multiple experimental datasets demonstrates that SPEC can accurately identify the source of a gene signature as myeloid or lymphoid, as well as differentiate between B cells, T cells, NK cells and monocytes. Using SPEC, we predict that myeloid cells are the source of the interferon-therapy response gene signature associated with HCV patients who are non-responsive to standard therapy.
SPEC is a powerful technique for blood genomic studies. It can help identify specific cell subsets that are important for understanding disease and therapy response. SPEC is widely applicable since only gene expression profiles from total PBMCs are required, and thus it can easily be used to mine the massive amount of existing microarray or RNA-seq data.
- Systemic Lupus Erythematosus
- Systemic Lupus Erythematosus Patient
- Cell Subset
- Gene Expression Signature
- Enrichment Score
1 Background
Gene expression data from blood genomic studies are widely used for investigation of human disease [1]. Predictive gene signatures have been developed to carry out differential diagnosis of infectious diseases [2], identify specific disease states [3] and characterize the immune response to vaccination [4]. However, is some cases gene expression signatures from blood can be weakly expressed and highly variable [5]. The identification of these signatures is complicated by the fact that blood is a mixed tissue, composed of multiple cell subsets, so that differential expression profiles can reflect changes in cell subset proportions, changes in subset-specific gene expression or both. In cases where the relevant disease genes are expressed in a subset-specific manner, as has been shown for SLE [6], analyses based on mixed cell expression data are inherently limited since differential expression of genes in one cell subset (e.g., monocytes) will be diluted by RNA from other cells. Experimental studies that isolate specific cell subsets before expression profiling can provide important biological insight by demonstrating subset-specific gene expression as well as increased predictive signal [7]. Given the large number of potential cell subsets that can be defined, the ability to identify the most informative subset(s) to isolate would be a great aid to these studies.
Genome-wide expression measurements based on total PBMCs reflect both condition-specific gene expression as well as the proportion of different cell subsets in the sample. Microarray deconvolution methods have been developed to take advantage of this latter dependence in order to quantify the relative proportion of different cell subsets [10]. In this approach, the expression level of each gene is modeled as a linear function of the expression from each cell subset comprising the sample. Deconvolution thus depends on prior knowledge of quantitative expression levels for each subset. In cases where the subset proportions have been measured experimentally, methods have been developed to compute subset-specific differential-expression for individual genes [11]. However, in addition to requiring measurements that are not available for many existing datasets, these methods do not directly address the problem of predicting the cell subset source of a pre-defined gene signature (e.g., one that is externally-derived or biologically-motivated). In some cases, clues to the cellular source of a gene expression signature may be found in the signature itself. This would be true when the signature is enriched for genes known to be expressed in a particular cell type [1], and may occur when the proportion of cells in the blood changes in the condition of interest. For example, Berry et al. found that changes in T-cell related genes included in a transcriptional signature of active tuberculosis resulted from changes in T cell number in the blood [3]. In cases where the predictive signature does not include a significant number of cell subset-specific genes, the problem may be addressed experimentally through the isolation of specific cell subsets followed by transcriptional profiling [7].
However, it is often not obvious which cell subset(s) will be the most informative. The time and expense of isolating and analyzing all possibly relevant cell subsets provides the motivation for the method developed here: a computational approach to predict the most likely cell subset source of a gene expression signature. The development of this method was also motivated by the clinical problem of predicting the outcome of antiviral therapy in patients chronically infected with hepatitis C virus (HCV). In this case, gene expression changes that are predictive when measured in hepatocytes are significantly blunted in PBMCs, making it difficult to develop highly predictive models from this cell population [5]. The isolation of specific cell subsets prior to expression profiling may allow for the identification of predictive signatures based on a blood sample, with significant advantages for the patient.
We have developed a computational method to predict the most likely cellular source for a pre-defined gene expression signature using transcriptional profiling data from total PBMCs. The general approach we propose is to: (1) estimate the relative proportion of each PBMC subset from individual patients using genome-wide transcriptional profiling data, and (2) correlate these proportions with the gene expression signature that predicts therapy. We theorize that the PBMC subset proportion that correlates most closely with the predicted gene expression signature over a large set of patients is the most likely source of the signal. As a specific example, let's consider a study where we find that Interferon Stimulated Genes (ISGs) are up-regulated in a subset of patients that fail to respond to a particular therapy. Now, we focus on the non-responding patients. If ISG expression tends to be higher in patients where the proportion of dendritic cells (DCs) is predicted to be high, then we would predict that DCs are the most likely source of the observed gene expression signature. Note that there can be two interpretations of this result with respect to the original study. First, the DCs may always express ISGs, but the proportion of DCs in the blood is higher for non-responders. Second, the DCs undergo specific gene expression changes in non-responders which cause the level of ISG expression to increase in the blood sample. In either case, DCs are the relevant population to follow-up experimentally. Here, we develop this general idea into a computational method called SPEC (Subset Prediction from Enrichment Correlation), which we then validate using both healthy and disease data. While the development of SPEC was motivated by the problem of predicting the response to therapy in HCV patients, the method is generally applicable to link gene expression signatures with specific PBMC subsets. Since only gene expression profiles from total PBMCs are required, the technique can easily be used to mine the massive amount of existing microarray data.
2 Results
- 1.
Estimate the relative proportion of each PBMC subset (e.g., B cells, T cells, etc.) across a population. We propose to accomplish this by calculating the enrichment score (E[s, i] for PBMC subset s and individual i) associated with a set of genes expressed specifically in each subset s, called a subset signature (subsetSignature[s]). This is summarized by the following pseudo-code:
N_P = Number of individuals
N_S = Number of PBMC subsets
exprData[i] = gene expression profile for individual i
subsetSignature[s] = list of genes representing PBMC subset s
querySignature = a list of genes input by the user
for each (i in 1 to N_P)
for each (s in 1 to N_S)
- 2.
Calculate the enrichment score of the query signature for each individual i in the population:
for each (i in 1 to N_P)
- 3.
Determine the Pearson correlation (across the population) between the enrichment scores of the query signature and each PBMC subset s:
for each (s in 1 to N_S)
- 4.
Predict the subset with the highest correlation to the query signature:
Predicted subset = s that gives rise to the max(C[s])
2.1 Estimating relative PBMC subset proportions using enrichment scores
The first step in our proposed method is to estimate the relative proportion of each PBMC subset. While a number of methods exist that can estimate these values, we chose to use enrichment scores calculated from cell subset-specific gene signatures for our initial implementation of SPEC due to its relative simplicity. As described in the methods, the enrichment score calculates the extent to which a gene signature is concentrated among the most highly expressed genes in a single genome-wide expression experiment. The cell subset-specific signatures are defined as groups of genes which have been found to be up-regulated in only a specific subset of cells in a mixed cell population. The resulting lists of up-regulated genes represent cell markers for that blood cell type, and include well known cell surface markers such as CD3 for T cells and CD19 for B cells. Lists of genes describing specific PBMC subsets have previously been defined in [12] and in [13]. Here, we choose to use signatures representing the B cell, T cell, NK cell, and monocyte subsets, We also included a signature for neutrophils despite the fact that they are not considered PBMCs, since neutrophil contamination can account for up to 20% of a PBMC sample [9]. In addition, we also use a more general set of signatures for the lymphoid and myeloid subsets (Figure 1). The lymphoid and myeloid signatures are composed of genes which are expressed in more than one cell type from those lineages, thus these subsets should vary according to the sum of all the cells in that lineage. We run SPEC separately on the detailed (B cell, T cell, NK cell, neutrophil and monocyte) and general (lymphoid and myeloid) signatures, as described below.
2.2 Validation of SPEC using split subset signatures
As an initial validation of SPEC, we sought to predict the source of a gene expression signature where the correct cell subset is known. This can be done by using our pre-defined cell subset expression signatures. For example, consider the B cell signature defined in [12]. The genes that are part of this signature can be split into two disjoint sets. One of these sets can be labeled as the cell subset signature and used to estimate the changes in the B cell population, while the other can be retained as the query gene signature being tested. A successful method would be able to identify this query signature as coming from B cells or from lymphoid cells. When running SPEC, we separately analyze the set of detailed signatures (B cell, T cell, NK cell, neutrophil and monocyte) and general signatures (lymphoid and myeloid), which represent cell types that are a superset of those covered by detailed signatures (Figure 1).
For each test, one of the subset signatures was randomly divided into two halves, with one half used as the query signature and the other half retained as the subset signature. This was repeated 200 times for each of the PBMC subset signatures. In order to determine the significance of the correlation value for each run we used a Monte Carlo permutation test (see Methods). The following two subsections describe the performance of SPEC on healthy and disease data.
2.2.1 Performance using healthy data
2.2.2 Performance in patients with disease
Performance of SPEC for linking split subset signatures using data from HCV patients
37 (69)
0 (30)
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14 (91)
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20 (56)
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22 (41)
3 (56)
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37 (66)
Performance of SPEC for linking split subset signatures using data from SLE patients
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2.3 Validation of SPEC using independently-derived signatures
2.3.1 Performance using independent subset signatures
Linking independently-derived cell-subset signatures
B Cells
T Cells
B Cells
T Cells
NK Cells
Myeloid Cells
2.3.2 Performance using experimentally-derived subset-specific disease signatures
2.3.3 Effect of query signature size on accuracy
2.4 Case study: identifying the ISG signature source for predicting therapy response in chronic HCV patients
The current standard therapy for chronic HCV consists of pegylated (PEG) Interferon (IFN)-α and ribavirin. However, this treatment is costly, poorly tolerated due to adverse side effects, and ineffective in many patients [22]. The majority of HCV infections in the United States are with genotype 1, which responds poorly to therapy (40% response rate) [23]. Methods to identify patients who are likely to be responsive to therapy would be a great help in clinical decision making. In addition, the ability to predict non-responsiveness could allow alternate treatments to be explored. Previous studies have found that patients who fail to achieve a sustained virologic response (SVR) after treatment display high baseline levels of IFN-stimulated gene (ISG) expression in hepatocytes prior to the initiation of therapy [24, 25], and the expression of these ISGs is not increased by IFN treatment. However, gene expression changes in total PBMCs in HCV patients appear blunted compared with looking directly at hepatocytes, making it difficult to develop highly predictive models from this cell population [5, 25]. Because there are significant advantages to using PBMCs for interrogating disease processes and performing clinical tests, it is important to investigate ways to improve the predictive power of PBMC gene expression profiling. We hypothesize that the accuracy of existing models based on PBMCs is limited because differential expression of ISGs is likely to be cell type-specific, and thus their signal will be diluted in total PBMC by RNA from other cells. This logic applies to non-ISG signatures as well. Therefore, using specific PBMC subsets for transcriptional profiling should improve our ability to extract knowledge from these data. Unfortunately, it is not obvious which cell subset(s) are the most informative, so we have applied SPEC to predict the most likely cellular source for predictive signatures in HCV.
We first generated a list of genes that were significantly up-regulated in patients classified as non-responders (compared with responders) based on gene expression profiling data from total PBMCs in 16 chronic HCV patients generated by Sarasin-Filipowicz et al. [5]. Using this gene list as the query signature, SPEC was then applied to the PBMC expression profiles from only the 6 non-responders in this study (see Methods) in order to predict the cellular source of the up-regulated genes. SPEC identified the myeloid subset (r = 0.74, p < 0.05), as the most likely source for this query signature. None of the associations with the detailed signatures reached statistical significance. This could be due in part to the relatively small number of samples used in the analysis. It is also possible that the therapy-response signature arises from multiple cell types in the myeloid family, or possibly from Dendritic cells (DCs), which were not included in our subsets for reasons previously described. Overall, these results suggest that gene expression analysis on the myeloid subset should improve our ability to predict therapy response in HCV patients.
3 Discussion
Blood is a mixture of many different cell types. The ability to associate a predictive gene expression signature with a specific cell subset would allow better classifiers to be developed through the experimental isolation of the most relevant cells. In addition, knowledge of which cells are being affected would provide important biological insights. No other computational methods exist to directly address this problem. In this study we developed a computational method, Subset Prediction from Enrichment Correlation (SPEC), to predict the most likely cellular source of a predictive gene expression signature. Our method requires only gene expression data from total PBMCs, and does not depend on knowing the subset proportions in the blood sample. SPEC uses enrichment scores to estimate the strength of gene expression signatures across individuals in a population. The PBMC subset signature showing the highest correlation with the query gene signature is predicted to be the source. We show that SPEC has good performance on both healthy and disease data, and correctly predicts the source of experimentally-derived subset-specific genes in SLE patients.
As a case study, we applied SPEC to transcriptional profiling data from chronic HCV patients in order to predict the potential source of a gene expression signature associated with non-responsiveness to standard therapy. Using SPEC, we predict that a stronger signal for non-responsiveness will be apparent when focusing on the myeloid subset. We hope that looking at the cell subpopulations separately will allow us to develop a blood test that can predict the clinical outcome of antiviral therapy prior to treatment.
The quality of cell subset gene expression signatures is critical to the performance of SPEC. It is clear from our results (e.g., sections 2.2.2 and 2.3.2) that some signatures are simply better than others. In this work, we use gene expression signatures for B cells, T cells, NK cells, neutrophils and monocytes available from [12] and [13]. We decided to exclude the DC subset signatures from [12] since we found that it did not work well within the SPEC framework. We speculate that this is due to the fact that DCs are such a small component of the total PBMCs (less than 1% on average [8]). More work is needed to define the qualities of a "good" signature for the SPEC framework. Other groups have proposed additional signatures that we plan to evaluate [26], with the long-term goal of defining a set of non-overlapping signatures for a detailed set of PBMC subsets. As an alternative to applying SPEC simultaneously to all available subsets, it is possible SPEC may be applied in a hierarchical manner: first predicting whether the query comes from the lymphoid vs. myeloid subsets then, if lymphoid, predict B cell vs. T cell vs. NK cell then, if B cell, predict naïve B cell vs. memory B cell vs... and so on.
An additional area for future investigation concerns alternatives to the enrichment score, which is currently used for estimating both the relative cell subset proportions and expression of the query signature. One possibility that we have already investigated is using microarray deconvolution, described in [10] and in section 2.1, to measure cell subset fractions. However, we found this method actually reduced the accuracy of SPEC (data not shown). Future work could explore other methods for estimating cell proportions from gene expression data [27, 28]. One thing to keep in mind is that SPEC does not require estimating the absolute proportion of all cell types within a single individual as provided by deconvolution methods, but rather assumes that we can estimate the relative proportion for each individual cell subset across a population. By trying to solve a harder problem, deconvolution methods may actually introduce more error compared with enrichment scores. Another advantage of enrichment scores is that they are likely to be less dependent on sample source (e.g., PBMCs vs. whole blood).
4 Conclusion
This work describes the implementation and validation of a computational methodology to support blood genomics studies with general applicability to basic and translational research. SPEC predicts the cellular source of predictive gene expression signatures. When applied to data from chronic HCV patients, SPEC predicts that focusing on myeloid cells may enrich for an interferon signature that has been observed in hepatocytes from patients that do not respond to standard therapy. The approach requires only transcriptional profiling data from total PBMCs, allowing wide applicability, and should improve the predictive power of blood gene expression profiling studies by allowing experiments to focus on the most informative cell subsets. It is also easy to envision how this framework could be combined with classification methods to define predictive signatures that are more likely to be cell subset-specific, and therefore amenable to experimental signal enrichment through sorting.
5 Methods
Microarray data and normalization
Raw Affymetrix microarray data were downloaded from GEO. Transcriptional profiles from PBMS were obtained for healthy controls (GSE11057, GSE11761, GSE14642, GSE15645, GSE7753, GSE8507), SLE patients (GSE11908), and chronic HCV patients (GSE7123, GSE11342, GSE11190). Cell specific transcriptional profiles for B cells, T cells and myeloid cells from SLE patients were obtained from GSE10325. These data were normalized with GCRMA [29] using the BioConductor software package in R [30].
Generation of SLE cell specific disease signatures
Differentially-expressed genes were determined for each cell subset (B, T and myeloid cells) by comparing SLE and healthy individuals using the LIMMA package in BioConductor [31]. Genes were separated into groups of upregulated and downregulated genes, ranked by FDR, and the top 100 genes were pulled out. Cell specific disease signatures were created by using only the genes unique to each signature.
Generation of HCV response signature
Gene expression data was obtained from the study by Sarasin-Filipowicz et al. [5]. Differentially-expressed genes were determined by comparing non-responders vs. responders using the LIMMA package in BioConductor [31]. Significance was defined using a false discovery rate cutoff of 0.1 using the method of Benjamini and Hochberg.
Enrichment score calculation
The enrichment score is calculated as described in [21] with p = 1 and weighted by the gene rank. Genes are ranked based on their expression values from a single transcriptional profile (highest to lowest expression). The enrichment score is the maximum deviation from zero of a running sum statistic which is calculated by walking down the ranked list of genes. The sum is incremented whenever a gene in the signature is encountered and decremented if the gene is not in the signature. Increments and decrements are weighted so the statistic sums to 0 over all the genes.
Microarray deconvolution
Deconvolution was performed to estimate the fraction of each cell subset from a single PBMC transcriptional profile using the methods described in [10]. The cell subset expression data used for deconvolution was downloaded from the supplementary material. The predicted cell proportions of related subsets were added together to generate the final cell fraction (e.g., CD4+ + CD8+ + T cell = Proportion of T cells).
Monte Carlo permutation test for P value estimation
To calculate a P value for the maximum correlation coefficient found by SPEC, we estimated the distribution of these values under the null hypothesis by randomly permuting the assignment of genes on the microarray to the query signature, while maintaining the size of the query signature. We then ran SPEC to determine the maximum correlation between the enrichment scores for these new queries and each subset signature. This process was repeated 1000 times to generate a distribution of maximum correlations. P-values were then estimated by fitting a normal distribution to these data and calculating the cumulative density for each SPEC result.
Implementation and Availability
SPEC was implemented in the R statistical programming language, and is available at http://clip.med.yale.edu/SPEC. Additional requests can be made by contacting CRB or SHK.
We thank Michael Robek for helpful discussions regarding the HCV data. We also thank the Yale High Performance Computing Center (funded by NIH grant: RR19895) for use of their computing resources.
Funding: The work of CRB and MU was supported in part by NIH Grant T15 LM07056 from the National Library of Medicine. SHK and MU were partially supported by a pilot grant from USPHS P30-DK34989.
Authors' Affiliations
- Chaussabel D, et al.: A modular analysis framework for blood genomics studies: application to systemic lupus erythematosus. Immunity 2008, 29(1):150–64. 10.1016/j.immuni.2008.05.012PubMed CentralView ArticlePubMedGoogle Scholar
- Ramilo O, et al.: Gene expression patterns in blood leukocytes discriminate patients with acute infections. Blood 2007, 109(5):2066–77. 10.1182/blood-2006-02-002477PubMed CentralView ArticlePubMedGoogle Scholar
- Berry MP, et al.: An interferon-inducible neutrophil-driven blood transcriptional signature in human tuberculosis. Nature 2010, 466(7309):973–7. 10.1038/nature09247PubMed CentralView ArticlePubMedGoogle Scholar
- Querec TD, et al.: Systems biology approach predicts immunogenicity of the yellow fever vaccine in humans. Nat Immunol 2009, 10(1):116–25. 10.1038/ni.1688PubMed CentralView ArticlePubMedGoogle Scholar
- Sarasin-Filipowicz M, et al.: Interferon signaling and treatment outcome in chronic hepatitis C. Proc Natl Acad Sci USA 2008, 105(19):7034–9. 10.1073/pnas.0707882105PubMed CentralView ArticlePubMedGoogle Scholar
- Hutcheson J, et al.: Combined deficiency of proapoptotic regulators Bim and Fas results in the early onset of systemic autoimmunity. Immunity 2008, 28(2):206–17. 10.1016/j.immuni.2007.12.015View ArticlePubMedGoogle Scholar
- Grigoryev YA, et al.: Deconvoluting post-transplant immunity: cell subset-specific mapping reveals pathways for activation and expansion of memory T, monocytes and B cells. PLoS One 5(10):e13358.Google Scholar
- Autissier P, et al.: Evaluation of a 12-color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans. Cytometry A 77(5):410–9.Google Scholar
- Schlenke P, et al.: Evaluation of a novel mononuclear cell isolation procedure for serological HLA typing. Clin Diagn Lab Immunol 1998, 5(6):808–13.PubMed CentralPubMedGoogle Scholar
- Abbas AR, et al.: Deconvolution of blood microarray data identifies cellular activation patterns in systemic lupus erythematosus. PLoS One 2009, 4(7):e6098. 10.1371/journal.pone.0006098PubMed CentralView ArticlePubMedGoogle Scholar
- Shen-Orr SS, et al.: Cell type-specific gene expression differences in complex tissues. Nat Methods 2010, 7(4):287–9. 10.1038/nmeth.1439PubMed CentralView ArticlePubMedGoogle Scholar
- Abbas AR, et al.: Immune response in silico (IRIS): immune-specific genes identified from a compendium of microarray expression data. Genes Immun 2005, 6(4):319–31. 10.1038/sj.gene.6364173View ArticlePubMedGoogle Scholar
- Palmer C, et al.: Cell-type specific gene expression profiles of leukocytes in human peripheral blood. BMC Genomics 2006, 7: 115. 10.1186/1471-2164-7-115PubMed CentralView ArticlePubMedGoogle Scholar
- Fall N, et al.: Gene expression profiling of peripheral blood from patients with untreated new-onset systemic juvenile idiopathic arthritis reveals molecular heterogeneity that may predict macrophage activation syndrome. Arthritis Rheum 2007, 56(11):3793–804. 10.1002/art.22981View ArticlePubMedGoogle Scholar
- Holland SM, et al.: STAT3 mutations in the hyper-IgE syndrome. N Engl J Med 2007, 357(16):1608–19. 10.1056/NEJMoa073687View ArticlePubMedGoogle Scholar
- Knowlton N, et al.: The meaning of clinical remission in polyarticular juvenile idiopathic arthritis: gene expression profiling in peripheral blood mononuclear cells identifies distinct disease states. Arthritis Rheum 2009, 60(3):892–900. 10.1002/art.24298PubMed CentralView ArticlePubMedGoogle Scholar
- Radom-Aizik S, et al.: Brief bout of exercise alters gene expression in peripheral blood mononuclear cells of early- and late-pubertal males. Pediatr Res 2009, 65(4):447–52. 10.1203/PDR.0b013e3181993473PubMed CentralView ArticlePubMedGoogle Scholar
- Radom-Aizik S, et al.: A brief bout of exercise alters gene expression and distinct gene pathways in peripheral blood mononuclear cells of early- and late-pubertal females. J Appl Physiol 2009, 107(1):168–75. 10.1152/japplphysiol.00121.2009PubMed CentralView ArticlePubMedGoogle Scholar
- Taylor MW, et al.: Changes in Gene Expression during Pegylated Interferon and Ribavirin Therapy of Chronic Hepatitis C Virus Distinguish Responders from Nonresponders to Antiviral Therapy. J Virol 2007, 81(7):3391–3401. %U [http://jvi.asm.org/cgi/content/abstract/81/7/3391] %U 10.1128/JVI.02640-06PubMed CentralView ArticlePubMedGoogle Scholar
- Taylor M, et al.: Cyclic changes in gene expression induced by Peg-interferon alfa-2b plus ribavirin in peripheral blood monocytes (PBMC) of hepatitis C patients during the first 10 weeks of treatment. Journal of Translational Medicine 2008., 6(1):Google Scholar
- Subramanian A, et al.: Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci USA 2005, 102(43):15545–50. 10.1073/pnas.0506580102PubMed CentralView ArticlePubMedGoogle Scholar
- Poynard T, et al.: Viral hepatitis C. Lancet 2003, 362(9401):2095–100. 10.1016/S0140-6736(03)15109-4View ArticlePubMedGoogle Scholar
- Zeuzem S: Interferon-based therapy for chronic hepatitis C: current and future perspectives. Nat Clin Pract Gastroenterol Hepatol 2008, 5(11):610–22.View ArticlePubMedGoogle Scholar
- Asselah T, et al.: Gene Expression and Hepatitis C Virus Infection. Gut 2009.Google Scholar
- He XS, et al.: Global transcriptional response to interferon is a determinant of HCV treatment outcome and is modified by race. Hepatology 2006, 44(2):352–9. 10.1002/hep.21267View ArticlePubMedGoogle Scholar
- Watkins NA, et al.: A HaemAtlas: characterizing gene expression in differentiated human blood cells. Blood 2009, 113(19):e1–9. 10.1182/blood-2008-06-162958PubMed CentralView ArticlePubMedGoogle Scholar
- Clarke J, Seo P, Clarke B: Statistical expression deconvolution from mixed tissue samples. Bioinformatics 26(8):1043–9.Google Scholar
- Erkkila T, et al.: Probabilistic analysis of gene expression measurements from heterogeneous tissues. Bioinformatics 26(20):2571–7.Google Scholar
- Wu Z, Irizarry RA: Preprocessing of oligonucleotide array data. Nat Biotechnol 2004, 22(6):656–8. author reply 658. author reply 658.View ArticlePubMedGoogle Scholar
- Gentleman RC, et al.: Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 2004, 5(10):R80. 10.1186/gb-2004-5-10-r80PubMed CentralView ArticlePubMedGoogle Scholar
- Smyth GK: Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol 2004, 3(1):Article3.PubMedGoogle Scholar
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