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# Laugh Love Quilt
*** A Cat Tale * On Sale
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2016 Row by Row
2016 Row by Row Oregon
2017 Row by Row
2018 Row by Row
30Æs Playtime 2018
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A Cat Tale
Ahoy Matey
All Stars by Tula Pink
Alphabet by Marcia Derse
Amber Reflections
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America The Beautiful
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Are We There Yet?
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Everything Kimberbell
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Give Thanks II
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Land That I Love
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March 201...
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Let it Snow
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Little Red Hen
Little Star in Flannel
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Longitude Rayon Batiks by Moda...
Looking Forward
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Machine Accessories
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Make Yourself At Home
Maxi-Lock Serger Thread
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Moon and Stars Flannel
Morris Garden
New May 201...
Morris Garden by Moda
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Newest Additions
Nicole's Prints
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Off We Go
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On Meadowlark Pond
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On The Chopping Block
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One Stitch at a Time
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Our Autumn Friends
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Paper Cuts by Cotton & Steel...
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Pin Stripe
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Pixie Basic Dots
Prairie Cloth by Moda
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Primitive Muslin Flannels
Project Dog Run
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Railways Express
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Road Trippin
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Sam & Mitzi
Santa Fe Wovens
New Jan 2...
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Silent Night
Silk Ribbons by The Thread Gat...
Simply Happy
Slow and Steady
Small Talk
Snow Day
Snow Happy
Solid Color 6-Strand Floss...
Solid Color Pearl Cotton No 8...
Spin Basics
Spirit of America
Spirit of America
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Spring Awakens
New Feb 201...
Stand Tall
New Dec 2018...
Stella Dots
Storybook Ranch
Sulky Blendables
Sun Kissed
New Feb 2019...
Sunday Supper
Sunflowers and Busy Bees
Sunnyside Up
New Dec 2018...
Sunshine Day
SusybeeÆs Kid Prints
Sweet Meadow New
Sweet Orchard
Sweet Things
Tabby Road by Tula PInk
The Ghastlies
The Reclaimed West
Tippy Toes
Torrid Zone - New Jan 2019...
Tribal by Free Spirit
New March 2019...
Twill Solids
Vanilla Icing * Grey Basics...
Varigated 6-Strand Floss
Varigated Pearl Cotton No 8...
Verona Velvet
Vintage Sports
Voysey 2018
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Waistband Elastics
Wanderlust By Hoffman
Watercolor Basics by Moda
We Whisk You a Merry Christmas...
Welcome Home Collection One...
Well Said by Moda
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Wild & Playful
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Main Menu
Class Schedule
Read about our
Class Policies
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class schedules.
Let's Explore My Design Center
Candle Mat
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Sku #:
1 session class with Carolyn
Class meets Wednesday, September 25, from 10:30 to 3:30.
Learn how to create your own embroidery designs with the My Design Center feature found on Brother Dream, Dream 2, Luminaire, and the new Stellaire machines.
We will be using both the line and region tools to create a pretty embroidery design.
As a bonus, I'll show you how to use the line tools to create applique.
to print your supply list.
class fee
We Whisk You a Merry Christmas Class
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5 session class with Carolyn
Class meets Thursdays, September 19, 26 and October 3, 17, and 24 from 10:30 to 4:30.
Capture a cozy, Christmas kitchen with gingerbread cookies, decorated aprons, hot cocoa, and baking treats in this delightful quilt. You will learn fun techniques with your embroidery machines that include working with chalkboard fabric, glitter vinyl, and more.
Students will need to purchase the book, embellishment kit and fabric kit from Cottage Quilts.
to print your supply list.
class fee
Basic Simple Maintenance
For Your Featherweight 221
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1 session class with Caryn Lee
Class meets Saturday, October 26 from 10:30 to 2:30
Have you purchased or been gifted with a 221 Featherweight? Is it sitting on a shelf because you are afraid to use it? Or do you use it but are not familiar with how to take care of it? Then this class is for you!
Come learn how to do simple basic maintenance on your 221. This class is taught in a casual and easy way so that you can learn to ōSimply Loveö your Featherweight! You will become confident in your ability to maintain your machine and not be afraid to use her. The material is offered in language that the everyday quilter/sewer can understand.
Tasks covered will include how to clean the gears, lubricate and oil your machine, treat the finish, learn to correct common mistakes to make your sewing experience easier, and much more.
Some history and facts will also be covered. Once you take this class you will not put your precious machine in a closet again but will use and take care of her confidently!
This class is not an advanced repair class!
to print your supply list.
class fee
CQ Sew Creative Software Club
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1 session class with Carolyn
Class meets Monday, October 14 from 1:00 to 4:00
This class is for Premier+2 Ultra customers.
I'll post a photo soon with all the details!
class fee
Garden Delights Blocks 1 and 2
Premier+2 Software Class
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1 session class with Carolyn
Class meets Monday, September 23 from 10:30 to 3:30
Garden Delights is a class for Premier+2 students who are also members of Vantage Software Support. We will be offering a class each month that will cover one or more Garden Delights lessons. It's a fun way to learn more about your software!
In this class we will be working on blocks 1 and 2.
Students need to print out the lesson and bring it to class. I suggest you start a binder for these lessons to stay organized. And, of course, you will need your lap top, power cord, mouse with extra batteries, and a USB stick (just in case).
class fee
Journal Jacket Workshop
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1 session workshop with Carolyn
Class meets Monday, October 21 from 10:30 to 3:30.
This class is for students who currently own a Pfaff embroidery machine with Applique Creator and Shape Creator or a Viking embroidery machine with Design Applique and Design Shaping. We are including machines with different hoop capacities: 360 x 260 and 360 x 200. The design and size of your project will change according to the machine you own.
Learn these unique features while you create a journal jacket, book cover, or iPad cover.
The class fee is $45.00 plus a kit fee of $19.99. Your kit includes faux leather, cotton prints for the applique and lining, fusible fleece, and colored elastic all cut and ready to use when you come to class. The class fee will be waived if your machine was purchased at Cottage Quilts.
to print your supply list.
The fabrics shown will be used in your project. The bird fabric will be cut for your applique. You may not have a bird, but there are many interesting parts to this fabric. The black fabric with type will be used inside the journal. The main fabric will be a black faux leather. We have samples in the store if you need a closer look.
kit and class fee
Kids Projects: Coin Purse
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Ages 8 to 14
1 session class with Jenny
Class meets Saturday, October 19 from 9:00am to 11:00am.
This class is for students who have completed Jenny's Basic Sewing for Kids classes. Or, for students who understand the basics of their machine: how to wind a bobbin, thread the machine, select a stitch, and sew forward and in reverse.
Now that students have learned the basics, Jenny is offering a series of classes building on those skills.
Pick and choose the classes you wish to attend.
to print your supply list.
class and kit fees
Kids Projects: Library Card
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Ages 8 to 14
1 session class with Jenny
Class meets Saturday, November 16 from 9:00am to 12:30pm.
This class is for students who have completed Jenny's Basic Sewing for Kids classes. Or, for students who understand the basics of their machine: how to wind a bobbin, thread the machine, select a stitch, and sew forward and in reverse.
Now that students have learned the basics, Jenny is offering a series of classes building on those skills.
Pick and choose the classes you wish to attend.
to print your supply list.
class and kit fees
Kimberbell Retreat
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5 day retreat meeting Monday through Friday, November 11 - 15, from 10:30 to 4:30.
Join us for five days of fun working on a Kimberbell machine embroidery quilt. Students can choose a Kimberbell quilt project or Bench Pillow.
The event fee includes instruction and lunch on us on Friday, November 15. BYOL, Bring your own lunch Monday - Thursday, or visit one of our local restaurants.
Students will need to purchase the book of embroidery designs and the kit from Cottage Quilts to attend this class. Some projects also have an embellishment kit.
Kimberbell designs include so many different techniques: working with mylar, glitter vinyl, free standing embellishments, fringe embroidery, chenille embroidery, and more. It's the perfect way to learn your embroidery machine and have fun at the same time.
All brand machines are welcome to attend this class.
The Supply List for each project will vary. Refer to your pattern instructions for the type of stabilizer, thread list, and miscellaneous items that may be needed such as Flex Foam or Shape Flex.
You will need to transfer the designs you will be using to a USB stick before you come to class.
class fee
Premier+2 Owner's Class
Lesson 3, Part 1
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This class is for Premier+2 Embroidery software = Levels Extra and Ultra
Class meets Monday, October 14 from 10:30 to Noon.
This month students will be covering the following topics:
Express Design Wizard, Encore, Decoration Tab, Photo Stitch, and Quilt Block Wizard
This Owner's Class is for students who purchased Premier+2 software from Cottage Quilts.
Rail Fence Quilt
A Beginning Class
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Sku #:
3 session class with Jenny
Class meets Wednesdays, October 16, 23, and 30 from 10:30 to 1:30.
Have you always wanted to learn how to make a quilt? Join our class with Jenny.
She has divided this class into three sessions. In the first class you will learn cutting with a rotary cutters and rulers, plus basic strip piecing. In the second class you will do basic construction of the quilt top. In the third class you will finish building your quilt top and start the quilting process.
This class is great for the absolute beginner or students that want to brush up on their quiltmaking skills.
to print your supply list.
class fee
Scan'n Cut SDX225 and New Disney
Owner's Class
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Sku #:
1 session class with Janet
Class meets Tuesday, October 1st from 10:30 to 12:30.
This is an owner's class for the new Scan 'n Cut SDX225 or SDX230D purchased from Cottage Quilts. Students will learn the basic operation of their new Scan 'n Cut machine.
to print your supply list.
Winter Village Tablerunner
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Sku #:
3 session class with Janet
Class meets Tuesday, October 15, 22, and 29 from 10:30 to 2:30
Join Janet for this fun class recreating a snowy winter village scene with snowmen, snowflakes, and snow-covered trees and houses.
The houses, sky, and snow backgrounds are pieced. The houses, snow people, trees, and snowflakes are fusible applique.
The pattern includes downloadable SVG files if you have a cutting machine. Or, if cutting your own applique with scissors, there is a traceable pattern.
In the first class, students will be preparing their applique fabrics either with a cutting machine or with scissors. In the second class, you will start piecing the backgrounds. In the third class you will iron on your appliques and start stitching them with a variety of stitches.
to print your supply list.
Be sure to do your homework before you come to the first class!
This class does not teach you how to use your cutting machine.
class fee


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I create things of value and serve clients thoughtfully, honestly and artistically.
Branding for small businesses
You don't know what you don't know.
Entrepreneurs and startups undermine the value of brand strategy: the mission, vision and values. It is the backbone of the brand, the funnel you run every decision through.
In this process, I'll guide us through a brand DNA exercise, marketing activity, logo concepting, and brand traits development.
Print + Digital Collateral
Simply put: This is formatting — laying out images and text together in a harmonious way across multiple compositions: an ad, a brochure, a magazine, a post, a flyer, a presentation, etc.
...always making sure to leave plenty of white space.
Powerpoint DESIGN + clean up
Need a presentation but don't know how to tell your story or have time to build slides?
I will clean up your slides or build a deck from scratch.
The best presentations contain minimal content while aligning a strong talk track.
Infographics + Illustration
Fonts, icons, and good layout. Let's speak to your audience visually.
It's important to find an interesting way to showcase your data or story.
I'll turn your spreadsheet data into something energetic and easy to read.
Creative direction For projects
It's all about how you tell the story and I'm filled with ideas. I learn visually and speak visually.
Maybe it's a lawn sign that serves to raise money, or a Facebook Ad Campaign, or a promotional video with voice over — whatever you're up against, together we'll create a theme, fine-tune the right language, and implement strong visuals to tell a damn good story.
everything else
Content development.
Video and photography.
Web development.
I partner with many local creative vendors for services outside my toolbox. These talented contractors have excellent portolfios, serving everyone from small businesses to corporate giants.

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Brid O' Connor
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Follow the Camino provides tours to Spanish Santiago de Compostela (St. James Route) since 2007 as the first travel operator for this pilgrimage place. Based in Dublin, Ireland. Offers perfect walking tours for groups and individuals. Book your tour
Click Offices provide a free search and support service for people searching for Serviced Offices globally. We offer you shared space offices across Dublin,Ireland, and multiple cities in the UK. Visit our website to see all of our locations.
MortgageLine helps you to find a great deal on a mortgage and life insurance. Our goal is to ease the complicated process of finding and applying for a mortgage and make it as simple as cost-effective as possible.
Hotel Woodstock is located in Ennis, Co. Clare, and offers 67 spacious rooms, cosy pub and beautifully landscaped gardens. It is a perfect place for a gateway for families and friends. Enjoy our hotel restaurant, spa or golf area.
The House Hotel is a 4 stars boutique hotel located in the Galway City Centre. The recently renovated hotel offers luxury accommodation for individuals, groups, families and business meetings. We also host all party occasions and events.


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The Senator representing Kogi West, Dino Melaye mocked Senator Godswill Akpabio as the 8th National Assembly tenure comes to an end.
Senator Godswill Akpabio lost his senatorial seat of Akwa Ibom North-West District during the February 23 general election.
At the wrap of the 8th Senate, Senator Dino Melaye in his valedictory message. He said:
#video:— Olayinka Samuel (@_olayinka) June 7, 2019
"Godswill Akpabio as a retired Senator, I will miss you in the 9th senate but we will definitely meet on the street of Abuja
~@dino_melaye pic.twitter.com/sZf03UY5oK


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Bradford film school opens thanks to Indian partnership
A new Bradford film school has opened its doors signalling the start of a new partnership between Bradford College and an Indian film firm.
Industry chiefs were in Bradford for the red carpet launch of the facility which will offer students the chance to experience the world of Bollywood.
The College has partnered with Mumbai-based television and film company, Whistling Woods International - and president Meghna Ghai-Puri flew in for the event.
She said: "We believe with Whistling Woods International and Bradford College we can provide a platform for students to join the global film industry. We have found the perfect partner in Bradford College."
Steve Abbott, an Honorary Fellow of Bradford College, said that the launch of the film school was a 'proud day for Bradford.'
He added: "As the UNESCO City of Film we always said the emphasis would be on learning through film and today's launch well and truly ticks that box."
Film school director Trevor Griffiths said: "Our ambition is for the film school to be open to people of all ages and abilities to enjoy the wonderful facilities we will be putting in place. When you think Bradford, think film!"
The school will be hosted in an existing refurbished building which will be fitted with dark rooms and studios.
The UK is set to leave the EU on 31 October 2019. Are you preparing for Brexit? Complete the North East Growth Hub and Tees Valley Business Compass' 3-minute survey →


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text: Entertainment Boardwalk Empire to end with season five Steve Buscemi in "Boardwalk Empire." Photo Credit: HBO By IVAN PEREIRA [email protected] Updated January 9, 2014 7:26 PM Print Share fbShare Tweet Email Nucky Thompson's small screen days are numbered. HBO announced Thursday that "Boardwalk Empire" will end after its fifth season, which will premiere in the fall. The show's creator Terrance Winter told the Hollywood Reporter that he and studio agreed that the story was coming to a close. "After much discussion with my creative team and HBO, we've decided to wrap up the series after such a great run and look forward to bringing it to a powerful and exciting conclusion," he said. The show stars Steve Buscemi as corrupt Atlantic City official Nucky Thompson who was involved in organized crime during the Prohibition era. It has received huge acclaim and strong ratings since its 2010 debut. By IVAN PEREIRA [email protected] Share on Facebook Share on Twitter Comments We're revamping our Comments section. Learn more and share your input.


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Katherine Danziger Protagonist of the book. Katherine is almost 18 and a senior in high school who is getting ready for college. When she meets Michael, she falls in love with him. One of the novel's central plotlines is her decision to lose her virginity to him as well as the sexual relationship they share together.
Michael Wagner Boy who Katherine meets and with whom Katherine falls in love. He is a senior at another high school nearby. They meet at a New year's Eve Party, and their relationship developes from there. He nicknamed his penis "Ralph."
Erica Small Katherine's best friend, who provides her with emotional support owing to Erica's ability to see situations from a realistic point of view. She sees sex as a phsical act, not a romantic one, unlike Katherine.
Artie Lewin A boy who is friends with Katherine and Michael. He is not very confident about himself and questions his sexuality and wonders if he is gay. Erica breaks up with him at a party after he scared her by saying he will commit suicide, and two days later he attempts to, resulting in him not graduating from High School.
Sybil Davison Katherine's friend (and Erica's cousin) who later gets pregnant when having loveless intercourse because she wanted to have experience in giving birth. She put the baby up for adoption, and hoped that they would name her baby girl Jennifer.
Jamie Danziger Katherine's little sister. She is proficient in music, art, and cooking. She is in the seventh grade and looks a lot like Katherine. She used to say that she was jealous of Jamie, but it passed.
Roger Danziger Katherine's father, a pharmacist who owns two drug stores.
Diana Danziger (née Gross) Katherine's mother, a librarian. She gives Katherine sexual advice.
Hallie Gross Katherine's maternal grandmother, a lawyer and progressive liberal.
Ivan Gross Katherine's maternal grandfather, who had previously had a stroke (he had to walk with a cane and sometimes at a loss for words), and would have another one later in the book which would claim his life.
Theo An older boy who Katherine inadvertently falls for while working at a camp, which spells the end of her and Michael. He also started the masscre of calling her Kat, and others started calling her that, though she disliked it.
David A boy Jamie falls in love with, and started a relationship with.
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We hope Bali Basics turned out to be helpful to you. Now that you've figured out where you want to stay on your Bali holiday, we help you with the sights we saw in Bali & loved. The attractions below are tried & tested, & advocated (& not mentioned in any order of preference)! Beaches…… Continue reading What We Loved Seeing In Bali
Tag: sunset
Bali Basics
Before we headed to Bali, we had a lot of confusion about its geography & location. Was it an island? Was it a part of Indonesia? How big was it? Blame it on ignorance. And, there's no better antidote for ignorance than travel. Once we'd been there, many contacted us when they were planning their… Continue reading Bali Basics
Beat The Heat! – 2
A few folks reached out to us to know more about the three destinations we recommended in Part I to escape the Indian summer. Glad we could be of help! But, three destinations are inadequate for six months of the intense north Indian summer. So, we bring three more long weekend getaways from Delhi. All… Continue reading Beat The Heat! – 2
Beat the Heat!
Come April & the Sun starts its mercilessness on the hapless souls of the National Capital Region. Right till September, it becomes a matter of hot, very hot & unbearably hot. In these six months, at least one getaway is needed to cooler environs. Jeolikot morning enveloped in mist Aren't we thankful that the Himalayas… Continue reading Beat the Heat!


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Kinde man dies after waves pulled him underwater
Fiancé, children were able to get to safety
HUME TOWNSHIP — A 30-year-old Kinde man died Monday night after being pulled into deep waters by a wave current, Huron County Sheriff Kelly J. Hanson stated in a news release.
Just before 9 p.m. Monday, the sheriff's office and multiple emergency services were dispatched to a beach of the Bay Drive residential area, just east of McGraw County Park in Hume Township.
It was reported a man had been pulled out into deeper water by wave current.
"Unfortunately he ended up underneath the surface and never came back up," Hanson stated. "The victim had been in the water with his fiancé and her children, who too were taken by surprise of the current. Fortunately the children were able to be brought back to shore by their mother, who in turn called 911."
Arriving first responders consisting of deputies, Caseville police, Port Austin and Caseville fire departments organized and began a water search assisted by family and bystanders.
Roughly 20 minutes later, the victim was found and brought to shore, where police EMTs, a responding paramedic and a registered nurse anesthetist who lives nearby began life saving efforts.
This effort continued for over 40 minutes on the beach, as well as in Central Huron's Ambulance on the way to McLaren Thumb Region in Bad Axe. However, doctors ended up pronouncing the victim dead shortly after 10:30 p.m.
"Even though the outcome was certainly not what any of us wanted, from what I witnessed on scene, I can attest to several fireman, police officers and ambulance personnel genuinely trying their best to overcome what unfortunately happened," Hanson stated. "Beach-goers are asked to exercise extreme caution when swimming in rough water."
"Our sympathy goes out to the family," the sheriff added.


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United States
> Big Rapids
We have 4 hotels in Big Rapids.
Holiday Inn Big Rapids
1005 Perry Ave.
This modern hotel is a 10-minute walk from Gate Family Entertainment Center and 1 mile from Ferris State University.Subdued rooms have desks, flat-screen TVs with cable channels and coffeemakers; some also include microwaves, minifridges and whirlpools. There's also free WiFi and high-pressure showerheads in the bathrooms, while kids 19 and under stay free with an adult.
Quality Inn & Suites Big Rapids
1705 S. State Street
Adjacent to Ferris State University, this modern 2-story hotel is also 7.5 miles from Hungerford Lake recreation area and 15.2 miles from St.
Super 8 Motel Big Rapids
845 Water Tower Road
This simple budget hotel is a 6-minute drive from the White Pine Nature Trail and an 8-minute drive from Robin-Hood Airport.All rooms include en suite bathrooms and cable/satellite TV, plus microwaves, minifridges and coffeemakers. There's free WiFi in every room and kids 17 and under stay free with an adult.
Country Inn & Suites By Carlson, Big Rapids, MI
15344 Waldron Way
Off Route 131, this informal hotel is 3 miles from both Ferris State University and the Jim Crow Museum of Racist Memorabilia.Relaxed rooms feature coffeemakers, minifridges and microwaves, as well as flat-screen TVs and complimentary Wi-Fi. Suites add either whirlpool tubs or living rooms with pull-out sofas.
More on Big Rapids .
Big Rapids is a city in the U.S. state of Michigan. The population was 10,601 at the 2010 census. It is the county seat of Mecosta County. The city is located within Big Rapids Township, but is politically independent.


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Neat little Y2K countdown thingie for your putergreenspun.com : LUSENET : TimeBomb 2000 (Y2000) : One Thread
For those of you who just love to download things.....who want to know exactly how much time we have until 12 midnight on 1-1-2000....who just want to know it ALL (as do I, despite obvious mental constraints which indicate that'll happen when pigs grow wings)....
I went a-webcruisin a while back and found this little free thingie you can put on yer 'puter, will tell you down to the second what the countdown is. It's called "Countdown 2000". Downloaded and tried about ten programs of this type, actually; this one was the simplest to install/use etc. IMHO.
You can configure it to start up automatically when the 'puter starts up by putting a shortcut to it in your Startup folder in Win 95/98; or fire it up manually; whatever floats your boat. Several different display formats; weeks, months, days, hours, seconds. All show the time left down to the second, tho. (Assuming your computer clock is correct -- also have a freebie that'll calibrate your computer clock with the US Naval Observatory Atomic Clock....if you want that, gimme a holler.)
The link for Countdown 2000 is http://www.icon-stl.net/~riclark/cd2k.html. I have absolutely nothing to do with the people who offer it, just have enjoyed using it and figured you folks might too. (^_^)
-- John Howard (Greenville, NC) (email@example.com), October 01, 1998
I found a beautiful Y2K screensaver from Silkmoth, who are apparently some very clever and talented folks Down Under. Countdown data elements (i.e., months, days, hours, minutes, seconds) float about over an oasis-like scene. People who visit my office cannot help but see it. Recommended.
Please note: I have absolutely no connection whatsoever with Silkmoth, other than admiring their work and having been known to enjoy a "tube of Foster's" ever so often. G'day!
-- Mac (firstname.lastname@example.com), October 01, 1998.
Cool beans, Mac! free, and very cheaply upgradable too
-- John Howard (email@example.com), October 01, 1998.
Thanks! I've been looking for something like this. You might want to try using "Nailit" from http://www.pcworld.com/software_lib/data/articles/system/other.html to keep it "always on top."
-- Mike (email@example.com), October 01, 1998.
Holy moly Mike -- that's a great page....gonna download NailIt and about a hundred other things, from the look of it...
(I don't have an "inner child".....I have an "inner downloader"....)
-- John Howard (firstname.lastname@example.com), October 02, 1998.
Okay - this doesn't load on your computer. But it can sit on your computer...
Found a neat countdown clock in one of the latest JCPenny sale catalogs. It was 24.95 (I think). But it is portable...and does continue to work without grid power... not assuming ANYTHING ever happens with that.... ;)
-- j (email@example.com), October 08, 1998.


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- Demonstrate a commitment to integrity and ethics
- Show respect for and value all individuals for their diverse backgrounds, experiences, styles, approaches and ideas
- Listen to others for understanding
- Assume positive intent
We Have A Vision
Our vision is to delight the world through foods and brands that matter.
Our purpose is to nourish families, so they can flourish and thrive.
We are a company of dedicated people committed to upholding our founder's vision of a healthier world. We push beyond boundaries and across borders to create foods that help fuel the best in everyone everywhere.
We Live Our Values
Our values are part of our DNA. They guide us, and we live them every day as we work with our customers and business partners, in our communities and with each other.
Through integrity, accountability, passion, humility, simplicity and a focus on success, we have created an inclusive and diverse culture. Where ideas can blossom, people can thrive, and success can flourish.
Learn more about how we're living our Founder's values on our Corporate Responsibility site.
- Take personal accountability for our actions and results
- Focus on finding solutions and achieving results
- Actively engage in discussions and commit to decisions once they are made
- Involve others in decisions and plans that affect them
- Keep promises and commitments made to others
- Personally commit to the success and well-being of teammates
- Embrace the belief that all injuries are preventable and that achieving a healthy and safe environment starts with "me"
- Show pride in our brands and heritage
- Delight our consumers and serve our customers through the high quality of our products and services
- Promote a positive, energizing, optimistic and fun environment
- Promote and implement creative and innovative ideas and solutions
- Value, promote and fiercely protect our reputation
- Value openness and curiosity to learn from anyone, anywhere
- Seek and provide honest feedback
- Be open to personal change and continuous improvement
- Learn from mistakes and successes in equal measure
- Never underestimate our competition
- Seek to continually simplify and improve processes, procedures and activities
- Work across organizational boundaries/levels and break down internal barriers
- Deal with people and issues directly and openly
- Prize results over form
- Achieve results and celebrate when we do
- Help people to be their best by providing coaching and feedback
- Work with others as a team to accomplish results and win
- Have a "can do" (positive) attitude and drive to get the job done
- Make people feel valued and appreciated
- Make the tough calls
We Give Back
In our communities and around the world, you'll find Kellogg employees giving back.
From volunteering at a local food bank, to supporting global programs such as Breakfasts for Better Days™, and responding to humanitarian and disaster relief efforts, we have a responsibility to make this world a better place to live. When you join our team, you'll make our world a better place too.
You can read the latest stories on Kellogg's Social K blog.Learn More


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Semi-quantitative bacterial culture is the standard method to diagnose urinary tract infections (UTI), but bacterial growth rate limits diagnostic speed and it is unreliable when patients have been pre-treated with antibiotics. Metagenomics could increase diagnostic speed and accuracy by sequencing the microbiome and resistome directly from urine samples, bypassing culture. However, a semi-quantitative approach – as needed for diagnosing UTIs – has not been established.
Metagenomics was deployed to identify and semi-quantify bacterial presence indicative of UTI, predict antimicrobial susceptibility (AMR), and results were compared to semi-quantitative culture. Whole genome sequencing of the corresponding uropathogens was done for comparison. Analysis time and cost were tracked.
Forty-one consecutive urine samples underwent metagenomic analysis. All culture positive samples contained >200ng of DNA, suggestive of a threshold below which UTI could be ruled out solely based on DNA quantity. A semi-quantitative Diagnostic Index (DI) was created by multiplying the total DNA quantity by the relative abundance of uropathogens per urine sample. The DI allowed discrimination of UTI from non-UTI samples in all but 1 case. Metagenomic detection of AMR determinants correctly predicted the phenotype of uropathogens in 20 of 32 cases. The metagenomic work-flow was 31h and cost €116 per sample, but could be reduced to 4.5h and €5 for low-DNA-yield non-UTI samples.
The genomic determinants of AMR and their distribution across uropathogens need to be better understood for prediction of AMR phenotypes by metagenomics. The introduction of the DI demonstrates the potential of semi-quantitative metagenomics to replace culture as rapid diagnostic method for UTI.
Urinary tract infections (UTIs) are amongst the most common infections that require antibiotic prescription, and in catheterized and hospitalized patients UTIs are the most common nosocomial infections(1-3). The high incidence of UTIs, paired with increasing resistance to first and second line antibiotics for common uropathogens, such as Escherichia coli, stress the need for rapid diagnostics in aid of appropriate antimicrobial treatment(4). However, the current standard for diagnosing UTIs is time-consuming as it is based on (semi-)quantitative culture of urine, followed by identification and antimicrobial susceptibility testing (AST) of isolated pathogens, which usually takes 1-3 days and for some pathogens up to 7 days. In addition, growth of clinically relevant bacteria may be hampered by prior antimicrobial treatment, which is common practice worldwide, resulting in diagnostic challenges and risks of misdiagnoses.
Dependence on culture for UTI diagnosis can be bypassed by quantitative molecular detection of uropathogens in urine specimens. Multiplex real-time PCR-based methods have shown promise for this purpose but are hampered by exclusive detection of selected micro-organisms and lack of, or incomplete information on antimicrobial resistance (AMR) patterns(5, 6). Direct metagenomic sequencing of microbial communities in urine provides opportunities for unbiased detection of uropathogens, including the presence of AMR genes, while current next generation sequencing (NGS) methods and bio-informatic tools facilitate generation of such results in a timely fashion. Metagenomic approaches have indeed shown promise in identifying uropathogens and their antimicrobial susceptibility patterns in urine samples(7). However, semi-quantitative detection of uropathogens is essential in order to replace culture-based diagnostics by metagenomic approaches since international clinical guidelines use bacterial loads (e.g. < 103 or 105 CFU/mL) to define evidence of UTI in different patient populations(8, 9). To facilitate such semi-quantitative metagenomic diagnostics for UTI, we developed and tested a metagenomic sequencing-based algorithm for determination of clinically relevant levels of bacterial DNA, indicative of urinary tract infections. Results were compared to the reference test semi-quantitative urine culture followed by species identification by MALDI-TOF, and automated AST. Whole genome sequencing (WGS) of isolated uropathogens served as benchmark for expected AMR gene content in metagenomes. We recorded the time to result and direct costs. The aim of this study was to provide proof of concept for the use of metagenomics as a tool for detecting UTI and predicting antimicrobial susceptibility patterns.
Urine Samples
During two days, all urine samples obtained from patients with suspected UTIs that were assessed at the clinical microbiology laboratory for culture, identification, and AST, were included in the study. We collected up to a maximum of 20ml of surplus urine. Urine samples, corresponding routine culture plates and AST results were collected and anonymized, according to local and national ethical requirements for diagnostic studies using surplus routine care patient samples.
Urine Culture
Urine was cultured according to local standard operating procedures (see supplementary data for summarized SOP). Bacterial isolates were identified morphologically and by MALDI-TOF MS (MALDI Biotyper, Bruker, Karlsruhe, Germany) and were reported in a semi-quantified way as the number of colony forming units (CFU) per ml.
A culture-positive urine was defined as growth of clinically relevant bacteria at numbers >103 CFU/ml and for the purpose of this study all culture-positive urine samples were assigned to the UTI group. Growth of 103 CFU/ml or less was reported as "no significant growth", while mixed growth of commensal vaginal, rectal, or mucocutaneous bacteria in any numbers or absence of any growth were reported as "commensal flora" or "no growth", respectively. The latter three culture results together formed the non-UTI group. The clinically relevant bacteria from culture-positive urines were subcultured on Columbia agar plates with 5% sheep blood (Biomérieux, Marcy-l'Étoile, France) prior to DNA extraction and WGS.
Antimicrobial susceptibility testing
AST was done directly on urine by disc diffusion if Gram staining of the urine revealed the presence of abundant bacteria of a single morphology, or on identified bacterial isolates using VITEK 2 (version 06.01; Biomérieux), both according to EUCAST guidelines and breakpoints(see Supplementary data, Summary of SOPs)(10).
DNA extraction and sequencing
DNA was extracted from 20 ml of urine. If <20 ml of urine was available, the sample was supplemented with PBS. The urine was first centrifuged for removal of human cells (2000 g, 30 s), followed by centrifugation of the supernatant for pelleting of bacterial cells (8000 g, 10 min). The bacterial pellet was pre-lysed with an in-house enzyme cocktail comprising achromopeptidase, mutanolysine, lysostaphine (1000:100:3) and lysozyme (1mg/ml) (Sigma-Aldrich, St. Louis, MO, USA) in TE buffer. This was followed by lysis using proteinase K and an in-house lysis buffer (Sodium-docecyl-Sulphate (1 %), Tween-20 (0.5 %) and Sarkosyl (0.5 %) in TE-buffer), after which automated DNA extraction was performed immediately using the NucliSENS easyMag platform (Biomérieux) following manufacturer's instructions. DNA from the cultured bacteria was extracted with the Wizard Genomic DNA Purification Kit (Promega, Madison, Wisconsin, USA). The Qbit dsDNA HS Assay Kit (ThermoFisher, Waltham, Massachusetts, USA) was used to measure DNA concentration. The Ion Xpress™ Plus Fragment Library Kit (Thermo Fisher Scientific) was used for PCR-free, manual library preparation according to manufacturer's specifications. Library quantification was performed with the Ion Library TaqMan Quantification Kit (Thermo Fisher Scientific). DNA was sequenced on the Ion Torrent Proton platform, set to produce an average of 1 million 200 base-pair length single-end reads per sample. Trimmomatic was used to remove low quality reads (settings WGS: headcrop 15, sliding window 4:20, Phred score 15; settings metagenomics: headcrop 15, crop 270, sliding window 4:20, Phred score 10, minlen 50)(11).
Metagenomic analysis
Human sequences in the metagenomic datasets were removed with Deconseq version 0.4.3, human genome version 37(12). The remaining reads were analyzed for identification of microbial DNA at subspecies level and determination of the organism's relative abundance using the CosmosID bioinformatics software package (CosmosID Inc., Rockville, MD)(13-16). A cloud version is accessible at https://app.cosmosid.com. The relative abundance of each bacterial organism per sample was expressed as a percentage of the total number of bacterial reads belonging to that organism, normalized for organism-specific genome length. Reads identified as eukaryote, viral, or archaeal were excluded.
All bacteria identified by metagenomics were classified as commensal or uropathogen. Commensal bacteria comprised members of the genito-urinary tract and skin microbiota generally not considered as urinary pathogenic. Uropathogens comprised all Enterobacteriaciae, other species known as common causative agents of UTI and putative uropathogens that have been described to cause UTI in rare cases(17). A full list of bacterial species detected and their designation is provided in the Supplementary data, table 1.
Finally, the resistome i.e., the pool of resistance genes present in the microbial community, was characterized from the metagenome with the CosmosID (CosmosID Inc., Rockville, MD) metagenomic software package using CosmosID's curated antibiotic resistance gene database. Resistome predictions were compared with results of AST and WGS of the corresponding isolated bacteria.
Whole genome sequencing
SPAdes/3.6.0 was used for read-assembly (settings: --iontorrent, -k 21,33,55 -- careful)(18). KmerFinder version 2.1 identified the bacterial species(19). If KmerFinder was inconclusive, a BLAST search of the NCBI database was done, using the assembled genome(20).
ResFinder version 2.1 was used with default settings (90% identity match, 60% coverage) for detection of acquired antimicrobial resistance (AMR) genes (https://cge.cbs.dtu.dk/services/ResFinder/ analysis date: 14th June 2017)(21). Additional acquired and chromosomal genes and mutations were detected from the assembled reads using the web-based "Resistance Gene Identifier" tool for searching the Comprehensive Antibiotic Resistance Database (CARD) using "strict" and "perfect" matches only with identity match of ≥99%(https://card.mcmaster.ca/analyze/rgi analysis date: 14th June 2017)(22). AMR genes identified by WGS served as benchmark of expected AMR gene content in metagenomic sequence data.
Metagenomic quantification
In order to integrate both DNA quantity and relative abundance in a single diagnostic measure, thus emulating the qualitative and quantitative properties of the reference test culture, we created the diagnostic index (DI). The DI is the product of the total quantity of extracted DNA in nanograms (D) and the relative abundance of uropathogens (RA) for each urine sample: DI = D * RA. Semi-quantitative culture and metagenomics results were compared one on one per urine sample for species identification and quantification. The median and interquartile ranges (IQR) for DNA yield, RA and DI for the groups UTI and non-UTI were compared. Thus, we assessed which metagenomic measure had best discriminatory power, displaying the least overlapping values between groups UTI and non-UTI.
Differences in relative abundance between uropathogens, as well as differences in total DNA yield between specimens were analyzed using the Mann-Whitney-U test (RStudio version 0.99.902). A significant difference was defined as a p-value <0.05. Graphs were designed with RStudio version 0.99.902. Sensitivity, specificity, and 95% confidence intervals (95% CI) were calculated using MEDCALC (https://www.medcalc.org/calc/diagnostic_test.php).
Culture and WGS of isolates
Forty-six consecutive urine samples collected for routine culture were included, results of which were reported as no-, commensal- or no significant (≤ 103 CFU/ml) growth for respectively 12, 19 and 4 urines, hence classified as the non-UTI group. Eleven samples were classified as culture-positive and constituted the UTI-group. From these 11 specimens, 15 uropathogens were identified in total: 2 Enterobacter cloacae, 6 Escherichia coli, 2 Klebsiella pneumoniae, 2 Staphylococcus aureus and 1 each of Proteus mirabilis, Morganella morganii, and Pseudomonas aeruginosa. Of these, 14 were available for WGS. Species identification by WGS confirmed identification by MALDI-TOF in all 14 cases.
Metagenomics of urine samples
Metagenomic sequencing and analysis was performed for 41 of the 46 included urine samples. In the UTI group, one sample did not contain surplus urine for metagenomic analysis, and DNA extraction was unsuccessful for another due to technical difficulties. In the non-UTI group, zero DNA was extracted from three samples for which culture showed no growth for one and commensal flora in 102CFU/ml for two samples. Thus, 9 UTI and 32 non-UTI urine samples, containing 5.7-1210.0 ng of DNA, were used for metagenomic sequencing and analysis.
Non-UTI group
The 11 sequenced urine samples yielding no growth in culture, showed at least one bacterial species to be present by metagenomics. In these samples, the most frequently detected genera were Gardnerella, Bifidobacterium, Enterococcus, and Lactobacillus (Supplementary data table 2-C). Of the 19 urine samples that grew commensal flora in culture, 17 were analyzed by metagenomics and the most frequently observed bacterial taxa were Bifidobacterium, Lactobacillus, Prevotella, E. coli, Staphylococcus epidermidis, and Gardnerella vaginalis (Supplementary data table 2-B). All four samples with no significant growth in culture, respectively showed E. coli, Bifidobacterium and Lactobacillus as most abundant taxa (Supplementary data table 2-D).
UTI group
In 9 of 11 urine samples from the UTI group that underwent metagenomic sequencing and analysis, 12 of the 13 cultured uropathogens were identified amongst the most abundant species by metagenomics, but some differences were observed (Table 1). Where S. aureus (sample 30), present in mixed growth with E. coli, was cultured at a concentration of 104CFU/ml, it was not found in the metagenomic dataset. Sample 19 showed Klebsiella pneumoniae dominance (90.4% relative abundance) with only 0.9% relative abundance of E. coli, whilst culture suggested equal growth of >105CFU/ml of both species. Similarly, culture of sample 49 suggested equal growth of Enterobacter cloacae and Pseudomonas aeruginosa, whilst metagenomics revealed E. cloacae dominance (relative abundance 53.9%) with P. aeruginosa present at 7.8% relative abundance. Additionally E. coli was detected at a RA of 29.4%. Only S. aureus (104 CFU/ml) was cultured from sample 8 whilst Klebsiella species was additionally detected by metagenomics (Table 1) in a patient pre-treated with trimethoprim. Metagenomics identified additional taxa compared to culture in 6 other UTI samples including Bifidobacterium species, Shigella species, Lactobacillus species, Bacillus cereus, Anaerococcus lactolyticus, Peptoniphilus harei and Atopobium vaginae.
Metagenomic quantification
The median relative abundance (RA) of uropathogens in urine samples from the UTI-group (93.6%; IQR 7.4) was significantly different (p<0.001) from that of the non-UTI group (0.3%; IQR 11.2) (Supplementary data, figure 1). The median DNA yield from urine samples of the UTI-group (642.4ng; IQR 456.5; range 203.5 – 1210.0ng) was and significantly different (p<0.001) from the non-UTI group (127.3ng; IQR 295.3; range 0.0-734.8 ng)(Supplementary data, Figure 2). Where the non-UTI samples had a variable DNA yield, all urine samples from the UTI group, yielded > 203 ng of DNA, forming a clear threshold below which no UTIs were found. However, both RA and DNA yield lacked discriminatory power as individual sample values were overlapping (Supplementary Data, figures 1 and 2).
Integrating RA and DNA yield per urine sample in the diagnostic index (DI) discriminated all 9 culture positive urine samples of the UTI-group from the remaining 32 non-UTI samples except for one. This sample corresponded to a culture showing no growth whilst metagenomics identified Aerococcus urinae as being present (RA 88%, DNA yield 374 ng) (Figure 1 and Supplementary Data, Figure 3. The median difference in DI for groups UTI and non-UTI was significant (p<0.0001), with only 1 overlapping value between groups. The sensitivity and specificity of the DI for correctly allocating a urine sample to the UTI or non-UTI group were respectively 100% (95%-CI: 66.4-100%) and 96.7% (95%-CI: 83.8-99.9%)(23).
Antimicrobial susceptibility
Phenotypic AST of the isolated uropathogens showed resistance against 0 to 6 drug classes per isolate, with resistance against an average of 3 drug classes per isolate, cumulating to 38 cases of AMR against a drug class for all 15 uropathogens together(Table 2, Supplementary data Table 3). Combined Resfinder and CARD analyses of WGS gave genotype-phenotype agreement in 22 of 38 resistant phenotypes.
Nine of 11 urine samples from the UTI group were available for metagenomic resistome analysis. These 9 samples contained 13 of the 15 cultured uropathogens, comprising 32 cases of resistance against an antimicrobial drug class, as per phenotypic AST. Metagenomic resistome analysis and phenotypic AST were concordant in 20 out of 32 cases (Table 2).
Five isolates (2 E. cloacae, 1 K. pneumoniae, 1 M. morganii and 1 P. mirabilis) derived from individual urine samples showed a nitrofurantoin resistance phenotype, but a nitrofurantoin resistance conferring gene was not detected, neither by metagenomic resistome nor by WGS analysis. Metagenomics did not detect a matching AMR gene to explain seven additional resistance phenotypes in five samples (Table 2) whilst AMR genes corresponding to these phenotypes were detected in the WGS assembly of the isolates.
In four samples, different AMR genes were identified by metagenomics and WGS of the corresponding isolate, although both could explain the phenotype (Table 2). In 2 of these samples (blaSHV and blaTEM genes detected by WGS in sample 19 and blaDHA genes in sample 45), coverage of the metagenomic datasets was too low to distinguish the highly homologous gene variants (5-10 SNP differences between identified genes) (Supplementary data, Figure 4). The third, a blaACT-28 gene detected by WGS in sample 7 was absent in the CosmosID database and was instead identified as the blaACT-9 gene (a close variant with 5 SNPs difference) in the corresponding metagenomic sample. Finally, in sample 30, containing E. coli phenotypically resistant to 3rd generation cephalosporins, metagenomic analysis identified an ampC gene whilst the blaCTX-M-27 ESBL-gene was identified in the WGS of the isolate.
Metagenomics outperformed WGS in three cases. In sample 28, containing E. coli phenotypically resistant to 3rd generation cephalosporins, metagenomics identified an ampC gene, whilst WGS detected blaTEM-1B, which encodes resistance against penicillins, but not 3rd generation cephalosporins. In two instances a resistance encoding genotype was identified by metagenomics but an equivalent coding gene was not detected in the corresponding WGS analysis. Both genotypes matched to a resistant phenotype: oqxA and oqxB coding for norfloxacin resistance in sample 45 and ampC for amoxicillin resistance in sample 23.
Cost and timing
The time from urine arrival at the laboratory to obtaining results by culture was approximately 24h for non-UTI samples and 48-72h for culture and AST for UTI samples. The cost of this analysis per sample was approximately €4 for a non-UTI and €10 for a urine sample from the UTI group, excluding cost of staffing.
Total time required for metagenomic analysis was approximately 31h per sample, which included 4.5h for sample centrifugation, DNA extraction, and DNA shearing. A total of 15.5h were required for library preparation. Four hours were needed for sequencing. The computation time for CosmosID analysis for all 41 datasets was 3.88 min per sample using a Linux analysis server with two physical processors, a total of 12 cores (non-multithreaded) and 64GB of memory. The cost of library preparation and sequencing was €116 per sample, excluding cost of staffing. Should a screening be made based on quantity of extracted DNA per sample, non-UTI samples would have a processing time of approximately 4h and a cost of €5, namely the cost of DNA extraction only.
This study demonstrates the potential of metagenomics for rapid detection of clinically relevant levels of uropathogens, thus distinguishing UTI from non-UTI urine samples.
Whilst several reports were previously published on the microbial community composition of urine(24, 25), successful metagenomic identification of uropathogens present in clinical samples has only been reported once to the best of our knowledge(7). However, that study did not include the semi-quantitative analysis required to establish a diagnosis of UTI(2, 3, 26). If metagenomics is to replace culture, a semi-quantitative analysis of clinically relevant bacteria present within the urine microbial community is critical to obtain a diagnostic test result, as has been done in this study.
Whilst all samples of the UTI group yielded >203.5 ng of DNA, 12 of 21 samples above this threshold were non-UTI urine samples, reflective of the fact that the procedure of urine sample collection is highly prone to bacterial contamination and that urine itself is not sterile(27). However, for none of the UTI samples was a low amount of DNA (<200 ng) associated with clinically relevant bacterial growth in culture, independent of the volume of urine from which DNA was extracted which ranged between 2 and 20 ml (data not shown). This observation suggests that establishing an unambiguous threshold of DNA extraction yield per urine volume is feasible due to the fact that in the presence of a UTI the bacterial DNA load in the urine increases exponentially. Thus, DNA quantity alone could serve as an initial screening step for a diagnostic algorithm, directing sequencing only those samples with a high DNA yield.
The total amount of time needed to detect and identify uropathogens and AMR genes in the UTI samples was significantly less (31h vs. 48-72hs) for metagenomics than for standard culture whilst metagenomics provided greater detail. Automated library preparation will further reduce processing time of UTI samples by several hours. Metagenomic costs were €116 vs. €10 for culture per sample. Although metagenomic sequencing is more expensive at the present time, sequencing costs are continuing to decline(28). At present €4 and 24h are required for ruling out a UTI by culture. Should a validated metagenomic diagnostic algorithm be in place, a UTI could be ruled out in a low yield DNA sample within 4.5 hours and costs would be reduced to the cost of DNA extraction – approximately €5 – making metagenomics highly competitive.
Overlapping values for the UTI and non-UTI samples were observed for both DNA quantity and relative abundance of uropathogens, indicating these parameters are unsuitable for identifying urine samples from patients with UTI. However, the DI discriminated UTI and non-UTI samples extremely well. The single outlier was a sample containing Aerococcus urinae in high relative abundance and high DNA yield, that had been reported to be culture negative, even though Gram-staining revealed Gram-positive cocci. A. urinae can be overlooked in cultures because of its morphological similarity to coagulase negative staphylococci(29), leading to false negative culture results.
Metagenomic analysis was successfully employed to identify uropathogens in all but two of the culture positive urine samples (Table 1). In the initial analysis Morganella morganii in sample 45 was not identified. The species was successfully identified after addition of the genome to CosmosID's reference database, highlighting the importance of well-curated and complete databases. In sample 30, S. aureus was cultured at a concentration of >104 CFU/ml. It is unlikely that insufficient DNA extraction due to poor lysis of Gram-positive bacteria can explain the apparent lack of S. aureus DNA, since an extensive pre-lysis protocol was used, although this cannot be ruled out(30). Alternatively, contamination of the agar plates may have occurred.
CosmosID metagenomic bioinformatics system identified AMR encoding genes that were confirmed to be present by pure isolate WGS analysis. However for a selected number of cases, metagenomic and WGS analyses identified a highly similar, but non-identical AMR gene. An explanation for this discrepancy can be the different databases used in the metagenomic and WGS analysis. For WGS, it has been shown that different bioinformatic AMR identification tools produce different results and that databases are often incomplete(31). In addition, low coverage of gene segments by metagenomic reads resulting in less accurate gene calling could explain differences between metagenomics and WGS. Interestingly, even though metagenomic and WGS methods identified different gene variants in those few instances, the variants were predicted to encode the same phenotype. However, accurate phenotype prediction on the basis of genomic information, whether WGS or metagenomics, remains a challenge. Phenotype clearly depends on more than the presence or absence of a resistance encoding gene. Whether antibiotic resistance mechanisms are expressed, depends on complex interplay between accumulating mutations or acquisition of resistance encoding genes and as yet unknown regulators, as well as environmental factors(32).
In conclusion, we provided a proof of concept of semi-quantitative metagenomic diagnostics for UTI, including the development of a "diagnostic index" based on uropathogen relative abundance and total DNA yield per sample, to facilitate rapid classification of urine samples to UTI and non-UTI groups. In future studies, clinical characteristics of patients and prior antibiotic use should be included to determine the sensitivity, specificity, and the positive and negative predictive value of this metagenomics analysis approach applied to urine samples using culture as the reference test.
V. A. J. wrote the manuscript. V. A. J., S. M. and M. E. J. performed the experiments. V. A. J., S. M., N. W., P. S. and N. A. H. performed the bio-informatics analysis. C. E. V. supervised the clinical diagnostic analysis. S. M., V. A. J., C. E. V., B. d W., R. R. C., M. D. d J., and C. S. designed the study. All authors provided critical review of the data and manuscript.
The authors would like to thank Cosimo Cristella for assistance in using RStudio and Frank Baas for critical input in sequencing planning and experiment design.
This study was supported by the COMPARE Consortium, which has received funding from the European Union's Horizon 2020 research and innovation program under grant agreement No. 64347. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Rita R. Colwell is Founder and Chairman of the Board of CosmosID®, a bioinformatics company, and some of the other authors are employees of the company. Affiliation with CosmosID does not alter the authors' adherence to all JCM policies as detailed in the online instructions for authors. Authors declare no conflict of interest with regards to this manuscript.


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At the recent PlayStation Experience it was revealed that Capcom has been working on the fifth iteration of the long running Street Fighter gaming series. In the past many years have gone by before a new entry was announced. This time the announcement of Street Fighter V comes on the heels of the final version of SFIV, Ultra Street Fighter IV. Not only that, but of the newest consoles, Ultra SFIV is making it's way to the PlayStation 4 in 2015.
As for Street Fighter V, everybody has heard that, as far as gaming consoles go, SFV will be exclusive to Sony's PlayStation 4. At least not everyone will be without as it will also see a release on PC. Further helping to lessen the blow of exclusivity is online "cross-play" with both the PS4 and PC platforms.
The response from the gaming community has been mixed, but not in the way you'd think. Everybody has been excitedly happy about the fact that a brand new entry in the Street Fighter series is upon us in the foreseeable future. However, backlash as come in the form of outrage that only the PS4 game console will get the game. No Xbox One release is planned. It has been rumored that this is because Capcom needed major funding for this project and Sony was willing given that it was exclusive to the PS4, of course.
Regardless of any rumored reasons for the business transaction taking place between Capcom and Sony, one thing is clear. SFV will only be on PS4.
The surprise to me isn't that such a deal would happen. It's that any gamers would be appalled by it. Exclusive third-party titles aren't a new concept, you know. It's been happening since the inception of the gaming industry. I think the reason for people being upset is the fact that we haven't had many third-party exclusives for a whole console generation. Before the PS3, Xbox 360, and Wii this was a normal sight to see. However, since 2005/2006 almost every single previously exclusive title from third-parties went multiplatform. When this happened then people complained. Now that they're used to it being that way they complain when a major release gets the old exclusive treatment.
I understand this hasn't been a thing for any Street Fighter game before. I also understand it sucks in general. I've had games I wanted to play in the past, but was denied because I didn't own the correct system. It sucks, but I moved on. You will too. I highly doubt Street Fighter V as a whole will be trapped on the PS4 forever. Sure they did say as much about SFV, but they didn't say anything about Super SFV.


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Event Guide
What to Bring
Don't forget your ticket! The link to your ticket is in your Etix confirmation email. To get scanned in, show your ticket on your smartphone or bring a printout. On-site printing is not available.
Bring your photo ID, if you'd like to enjoy alcoholic beverages. It's a good idea to pack lightly for the day. Restricted items include outside food or drinks, chairs, umbrellas, pets, blankets, and coolers. No professional photography, audio-recording equipment, selfie sticks, or GoPro handles are allowed.
No guns allowed. All bags subject to search. Smoking and e-cigarettes are prohibited from both the Walker and Minneapolis Sculpture Garden.
Bring an empty water bottle!
Bring cash or use our onsite ATMs. All food vendors accept cash; some will accept cards.
SATURDAY JUNE 29, 2019 | 1:00 pm Doors Open
|Main Stage
|Garden Stage
|2:30 pm deM atlaS
|3:45 pm Heart Bones
|4:35 pm The Beths
|5:15 pm X
|6:20 pm Adia Victoria
|7:00 pm Courtney Barnett
|8:05 pm Bad Bad Hats
|8:45 pm The National
|10:00 pm End
Food Vendors
Meet the makers behind Rock the Garden's drink sponsors! Enjoy complimentary samples and free swag from Summit Brewing Company, Sociable Cider Werks, and Tattersall Distilling from 1:30–2:30 pm in Esker Grove.
Free Hydration
Bring your empty water bottle and enjoy free, refreshing water throughout the Rock the Garden concert grounds, provided by Tap Minneapolis.
Food & Beverages
Food and beverages will be on sale from 1–10 pm. ID required to purchase alcohol. There will be many great food options available throughout the day and into the night. All vendors accept cash; some will accept cards.
|Best Way:
|Gyros, Chicken, Falafel, Pita Bread
|Esker Grove:
|Sandwiches, Grilled Vegetables, Donut Holes
|Foxy Falafel:
|Chicken Shawarma, Cauliflower Steaks, Hummus & Pita, Cheese Curds, Falafel Balls
|Free Indeed Foods:
|Gluten Free Mini Donuts, Gluten Free Chicken Bites, Smoothies, Lava Smoothies, Lemonade
|Fro Yo Soul:
|Frozen Yogurt
|Beer Cheese Nachos, Bacon Brats, Black Bean Burgers, Thai Chicken Wrap, Jerk Pork Flatbread, Watermelon Salad
|Gorkha Palace:
|Veggie Momos, Chicken Momos, Veggie Curry, Mixed Veggie Curry
|Hot Indian Foods:
|Indurrito, Rice Bowl, Indi Fries
|Kennedy Concessions:
|Hamburger, Cheeseburger, Bacon Cheeseburger, Veggie Burger, French Fries
|Cheese Curds:
|Fried Cheese Curds
|Que Viet:
|Egg Roll on a Stick, Garlic Cream Cheese Wontons, Bubble Tea, Vietnamese Iced Coffee
|R Taco:
|Grilled Fish Taco, Black Bean Taco, Fried Chicken Taco, BBQ Pork Taco, Gringo Taco
|Taulelle Corn Dogs:
|Foot-Long Corn Dog
|The Anchor Fish & Chips:
|Fish & Chips, Meat Pastie & Chips, Veggie Pastie & Chips, Battered Sausage & Chips, Poitin Poutine, Curry or Gravy Chips, Slaw, Beans, Curry, Gravy
|Wild Earth Pizza:
|Cheese Pizza, Pepperoni Pizza, Margherita Pizza, Sausage Supreme, Gluten-Free Crust
During the Festival
Enjoy cold drinks from Summit Brewing Company, Tattersall Distilling, and Sociable Cider Werks. Recharge with a variety of food and beverages. Shop the merch tent, stocked with Rock the Garden T-shirts and band merchandise.
Are you an MPR or Walker member? Thank you! Bring your member card and stop by the membership tent to pick up a special Rock the Garden gift as our thanks to you (limited quantities available).
Portable restrooms are available at the top of the hill and near the North Entry in the Minneapolis Sculpture Garden. Dedicated VIP restrooms are located off the rooftop terraces.
VIP offers a stellar view of the main stage from Walker's rooftop, the air-conditioned comfort of the Garden Terrace Room, and access to indoor restrooms. Your VIP ticket will get you exclusive access to our terraces and entry to the Grove, a viewing area near the main stage.
All food and nonalcoholic drinks in the VIP level are complimentary. You'll also receive 3 drink tickets, free parking in the underground ramp, and a Rock the Garden T-shirt. Other complimentary activities include a round of Skyline Mini Golf and video selfies in slow motion.
Atomic Data
Connect to the free #AtomicData RockTheGarden WiFi network and authenticate with Facebook, Twitter, or email. And be sure to stop by the Atomic Data tent for a chance to win free beer!
Autism Society of Minnesota
Pick up a drawstring backpack!
Planned Parenthood
Grab a fabulous pair of pink sunglasses and a one-of-a-kind bandana, then rock your new swag while you snap a selfie in front of the Planned Parenthood garden mural. Plus, learn more about how you can strengthen Planned Parenthood by becoming a patient. Don't miss it!
Robins Kaplan VIP
VIP guests can enter for a chance to win a FjallRaven backpack! The first VIP 250 visitors will get a free water pouch.
Sociable Cider Werks
Come visit the Sociable Cider Werks tent during Rock the Garden where you'll be entered to win VIP tickets to their summer concert. And while you're there, try your luck at the prize wheel to take home some Sociable swag!
Sparkling Ice
Stop by the Sparkling Ice Tropical Flavor Lounge to enjoy a complimentary bottle of Sparkling Ice and add to the giant coloring wall! Plus, don't forget to try the limited-time-offer mystery flavor! Guess the flavor on sparklingice.com/whattheflavor for a chance to win over 500 prizes.
Tattersall Distilling
Visit the Tattersall booth and play corn hole for a chance to win drink tickets, BOGO drinks in the Tattersall cocktail room, and Tattersall gift cards. There will also be giveaways including sunscreen and a raffle for a chance to win a Tattersall Bootlegger Yeti insulated jug! #tattersalldistilling
Timber Tech
Go Against the Grain at TimberTech's booth, featuring the melody of an immortal rock anthem built right into the deck itself. Come get your photo taken in front of the iconic Spoonbridge and Cherry, create your own boomerang video, and become part of a mural.
University of Minnesota
Stop by the #UMNgarden to snap a photo with Goldy Gopher, play a ball-toss game for giveaways, and test your knowledge of U of M trivia for a chance to see one of this year's Rock the Garden artists from backstage or in the photo pit. If you're hanging out in VIP, come see us for some extra sweet giveaways and U of M trivia fun!
Women Winning
Stop by the Women Winning booth to pick up your feminist swag and meet the Women Winning team!
Accessible for Everyone
All performances will have ASL interpretation.
We offer accessible viewing platforms for both stages. These areas have chairs for seating, in addition to space for wheelchairs.
Accessible all-gender, single-stall restrooms are located inside the Main Lobby of the Walker Art Center. There are also accessible portable bathrooms and water-filling stations in both areas of the Garden.
For parking accommodations or for any other access needs or questions, please email firstname.lastname@example.com or call 612-375-7564. If possible, please contact us 24 hours or more in advance of the event.
Accessible parking for Rock the Garden is located in our underground parking ramp. The ramp is located on Vineland Place. On the day of the event, it can be accessed by coming from the west or driving on Groveland Terrace and down the hill. To enter the garage, simply let the guard at the entrance know that you are using ADA parking. From the ramp, you can proceed into the event through the VIP gate, located in the hallway to the museum on the P1 parking level.
Seating Areas:
We will have wheelchair accessible seating areas that also include chairs by both stages. The ADA area for the Main Stage is just off the Esker Grove patio, by the main entrance to the museum. The ADA area for the Garden Stage is just north of that stage. You are welcome to use both of these areas along with anybody who is attending the event with you.
ADA port-a-potties are located throughout the event. You can find two accessible all-gender restrooms (single-stall, locking) in the corridor between the parking garage and the Main Lobby. More restrooms, including a family restroom, are located on the Walker's lower level, next to the Art Lab.
Food Allergies and Medication:
Outside food and beverages are not allowed in Rock the Garden. If you have a food allergy or need to bring medication into the event, we are happy to make an exception to this rule. Simply let security know as you are coming through the gate that you have a medical need and have been authorized to bring in your items.
Water Stations:
An accessible water station is located on Vineland Place. There are also water fountains located in the lower level bathrooms.
Quiet Areas and Baby Care Spaces:
If you need a private space to nurse, use a pump, or take a break, please visit the medical tent located in the Cargill Lounge near the Main Lobby. You are welcome to nurse anywhere in the event.
If you need to store any medical equipment during the event, lockers are located in the lower level just off the main lobby. Check in at the Main Bazinet Lobby desk to grab a key.
Paid event parking is available in the Dunwoody Institute lot for $10 in advance through Etix or for cash on the show day. The Vineland Place underground ramp is reserved for VIP ticket holders and closed to the general public.
Rock the Garden will be held rain or shine. However, staff reserves the right to cancel the event or part of the event due to severe weather. No umbrellas are allowed.
Other Information
This event will be filmed, photographed, or recorded. Ticket holders consent to appear in this documentation and its future use by the Walker Art Center, 89.3 The Current, and Rock the Garden sponsors.
Reentry is allowed with a valid wristband. Children ages 6 and older must have a ticket for entrance. No refunds; all ticket sales are final. Advance tickets are print-at-home and will be received by email; no printing is available on-site.
The Walker galleries and Walker Shop will be closed during Rock the Garden.
Going Green?
Metro Transit bus lines 4, 6, 12, and 25 serve the Walker. All ticket holders are eligible for a free ride on Metro Transit buses and Metro lines to and from the event. Click here to download the free pass.
Bike racks with parking for 500+ bikes and a Nice Ride bike sharing station will be located on Hennepin Avenue/Groveland Place and adjacent to the North Entry in the Minneapolis Sculpture Garden.
Rock the Garden is proud to be a zero-waste event.
Vendors & Press Contact
Interested in being a vendor or sponsor?
Contact Ben Geffen
Press requests:
Contact Rachel Joyce
Our Sponsors
The Walker Art Center and 89.3 The Current® present Rock the Garden 2019.
Lead Sponsors
Supporting Sponsors
Official Beer
VIP sponsor
Print Media Partner
The Minneapolis Sculpture Garden is a project of the Walker Art Center and the Minneapolis Park & Recreation Board.
Rock the Garden is proud to be a zero-waste event.


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Dream11 today match prediction for Ireland vs Zimbabwe 2nd T20 match playing on 12 July 2019. We provide IRE vs ZIM dream11 team fantasy cricket news, IRE vs ZIM match dream11 prediction, IRE vs ZIM dream 11 fantasy cricket team playing 11 and IRE vs ZIM match dream11 expert data. Get more winning tips and today prediction about Zimbabwe tour of Ireland 2019 in our prime page.
When & Where: The 2nd T20 match will be played at Bready Cricket Club, Northern Ireland. IRE vs ZIM dream11 leagues will be closed on 12th July, 8:30 PM.
IRE vs ZIM Dream11 Prediction | Ireland vs Zimbabwe Dream11 Team Preview
Ireland washed out the Zimbabwe ODI series by 3-0. Now, they looking to dominate three T20 Schedule.
The First T20 match abandoned without a ball bowled due to rain. IRE and ZIM going to play 2nd T20 in Northern Ireland.
Ireland Team Preview & Dream11 Prediction:
Paul Stirling and Kevin O Brien will open the innings. Both are very attacking batsmen but Kevin O Brien looked off-color in ODIs. Mostly he will do his best in T20 matches.
Most of the users will keep Paul Stirling as a captain choice from the Ireland team.
Andrew Balbirnie will bat at one down. He is like hit or miss character. He may score 50+ runs or may be dismissed for single-digit runs.
Lorcan Tucker is inconsistent with the bat. He scored a half-century in the 2nd ODI match.
Greg Thompson, Shane Getkate, and Mark Adair will handle the middle order batting.
George Dockrell is a very attacking batsman as well as a key bowler. Rankin and Joshua Little will assist him in the bowling unit.
Why Dream11 Prime? Check our latest dream11 prime results
Zimbabwe Team Preview & Dream11 Prediction:
Zimbabwe team is fully depending on two or three players. Others are totally out of form and giving poor performance.
Craig Ervine is an attacking batsman and consistent scorer for them. He scored three half-centuries and a century in the last 10 matches.
Brendan Taylor failed in ODIs but he is a good T20 batsman. So, he will be a good choice for vice-captain option.
We can try Hamilton Masakadza and Solomon Mire in GL teams.
Sean Williams and Sikander Raza will play a key role with both the bat as well as the bowl.
Kyle Jarvis and Chris Mpofu will lead the bowling unit. All-rounders will assist them.
IRE vs ZIM 2nd T20 Dream11 Tips To Win ALL Leagues – 12 July 2019
IRE vs ZIM Playing 11:
Ireland Predicted Playing 11: Paul Stirling, Andrew Balbirnie, Shane Getkate, Gary Wilson(w/c), Kevin O Brien, Boyd Rankin, Lorcan Tucker, George Dockrell, Joshua Little, Mark Adair, Greg Thompson
Zimbabwe Predicted Playing 11: Hamilton Masakadza(c), Ryan Burl, Craig Ervine, Brendan Taylor(w), Sean Williams, Sikandar Raza, Solomon Mire, Kyle Jarvis, Chris Mpofu, Tendai Chatara, Peter Moor
IRE vs ZIM Dream11 Key Players:
Ireland: Paul Stirling, Shane Getkate, Kevin O Brien, George Dockrell, Joshua Little, Mark Adair, Greg Thompson
Zimbabwe: Ryan Burl, Craig Ervine, Brendan Taylor, Sean Williams, Sikandar Raza, Kyle Jarvis, Chris Mpofu
IRE vs ZIM Dream11 Team Screenshot:
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