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New Pokémon GO Shinies Released In Celebration Of New Ingress Prime
Niantic, Inc., makers of the mobile game Pokémon GO, has released an update of their original game Ingress called Ingress Prime.
In celebration of this event, Niantic is releasing new shinies to Pokémon GO: the green Cubone and blue Ponyta. New items are also available in the Pokémon GO shop to commemorate Ingress.
Niantic has released new shinies to the game almost every week now with shiny Pinsir just concluded over the weekend. Look for more suprises before the year ends.
Via Press Release:
Niantic Unlocks the Mysteries and Intrigue of the Real World in INGRESS PRIME
It's Time to Move and Work Together with Friends in Niantic's New Real World Mobile Game, Available Now on the App Store and Google Play
SAN FRANCISCO – Nov. 5, 2018 – Niantic, Inc., creator of Pokémon GO, has launched Ingress Prime, a real world mobile game that transforms cities and neighborhoods across the globe into a vast science fiction universe ripe for exploration and adventure. Ingress Prime represents a pioneering intersection of real world mobile games and innovative massively multiplayer gameplay, uniting players worldwide to collaborate in order to drive the deep narrative forward. Ingress Prime is available as a free download on the App Store for iPhone and on Google Play for Android devices.
Ingress Prime builds upon Niantic's foundational real world augmented reality game, Ingress, and leverages cutting-edge location-based technology to reveal the global game board and uncover Portals disguised as public art and landmarks. Agents will begin their journey in Ingress Prime with an all-new interactive story to introduce players to the immersive sci-fi universe and gameplay mechanics required to capture these Portals and the powerful resource, known as Exotic Matter (XM), leaking from them. Upon completing the training, players must choose a side between The Enlightened and The Resistance, two factions battling for control of these landmarks and the fate of the world.
"Ingress Prime is built on Niantic's Real World Platform and serves as a key driver for innovation around gameplay and technologies for all Niantic products," said John Hanke, founder and CEO of Niantic. "Since its original release in November 2012, Ingress and its passionate and diverse user community have been an inspiration for the team to create new and innovative ways for players to work together both in-game and in the real world."
As with the original, the Ingress Prime storyline will evolve and change based on how Ingress Agents interact with one another and the outcome of Ingress real world "Anomaly" events. As Agents collaborate and compete, the story will expand through a new interactive hub (www.ingress.com/investigate) which serves as the entry point into the deep Ingress universe. A new Ingress web series, "The Dunraven Project," serves as an entry point, fusing narrative and real-world Agent achievements. The two Factions, the "Enlightened" and the "Resistance," are at the spine of the narrative and in Ingress Prime they come to life in a new way, as two functioning artificial intelligence entities, with which Ingress Agents and prospective Agents can interact. They are set to come online shortly after launch and will play a pivotal role.
Niantic's hallmark real-world "Anomaly" events will play an important role in Ingress Prime, putting the outcome of key narrative arcs into the hands of Agents. These Anomalies bring Agents from both factions together for a weekend of competition, exploration, and celebration. For Ingress Prime, Niantic plans to host 12 large-scale "Primary" Anomaly events (three each quarter in Asia, Europe, and the Americas), complemented by more than 100 community-driven "Satellite" Anomaly events. The new events kick off on November 17 in major cities around the world including Barcelona, Austin, and Hong Kong. A full listing of events can be found at www.ingress.com/events.
A whole new dimension of the Ingress world will be showcased in "The Ingress Anime," an all new CG-animated series developed in collaboration with Fuji TV and Craftar Studios in Japan. It will be released worldwide in 2019 on Netflix. The show merges Ingress Agents, gameplay, and narrative into a global sci-fi adventure thriller and features music from a number of artists including Alt-J.
Ingress Prime is available as a free download on the App Store for iPhone and on Google Play for Android devices. To learn more about Ingress Prime and receive future updates visit www.ingress.com and following www.twitter.com/ingress
About Niantic, Inc.
Niantic, Inc., builds mobile real world experiences that foster fun, exploration, discovery and social interaction. Originally incubated within Google, Niantic, Inc. spun out in 2015 with investments from Google, The Pokémon Company, and Nintendo, and raised an additional $200 million in financing in 2017 from Spark Capital, Founders Fund, Meritech, Javelin Venture Partners, You & Mr. Jones and NetEase, Inc.. The company's immersive Real World Games Platform powers the ground-breaking augmented reality games Pokémon GO and Ingress, which are available on the App Store and Google Play. Harry Potter: Wizard's Unite is Niantic, Inc.'s third game and is being co-developed with Warner Bros. Interactive Entertainment. For more information on Niantic, visit www.nianticlabs.com.
Press play to watch the video below.


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"A Christmas Memory" is a short story by Truman Capote. Originally published in Mademoiselle magazine in December 1956, it was reprinted in The Selected Writings of Truman Capote in 1963. It was issued in a stand-alone hardcover edition by Random House in 1966, and it has been published in many editions and anthologies since.
The largely autobiographical story, which takes place in the 1930s, describes the lives of seven-year-old Buddy and his elderly cousin Sook, who is Buddy's best friend. The evocative narrative focuses on country life, friendship, and the joy of giving during the Christmas season, and it also gently yet poignantly touches on loneliness and loss.
Now a holiday classic, "A Christmas Memory" has been broadcast, recorded, filmed, and staged multiple times, in award-winning productions.
This has to be my all time favorite Christmas film. An obscure gem that is hard to locate on video. A MUST SEE ~ This defines the true meaning of Christmas for me.
"It's fruitcake weather..."

subset: commoncrawl


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Summerby's Catering
At Summerbys Catering we know how important excellent food is and as a family-run business we also understand that it is essential for your special day to run smoothly. Let Summerbys Catering be a part of your big day with high quality service and great food which will always be remembered. With over 30 years of catering experience, Steven and his team have created different menu options which are perfect for any occasion at affordable prices.
Telephone: 01158 479 083
Telephone: 01158 479 083
Email: firstname.lastname@example.com
Website: http://www.summerbyscatering.co.uk
Summerby's Catering
3 Glen Helen


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Palace defends Pi?ol amid quit call
Malacanang defended on Thursday Agriculture Secretary Emmanuel Pi?ol, saying he was just misquoted on the issue of legalized smuggling of rice in Zamboanga.
Presidential Spokesperson Harry Roque issued the statement amid call by some senators and congressmen for his resignation for the rice crisis in Mindanao, particularly in Zamboanga City and island provinces of Basilan, Sulu and Tawi-Tawi (ZamBaSulTa).
"The recommendation of the Department of Agriculture (DA) is to establish a rice trading post where government can collect customs duty on legally imported rice with import permit from the National Food Authority (NFA)," he said.
He also said Pi?ol still enjoys the trust and confidence of President Rodrigo Duterte.
"Unless fired, yes," Roque said when asked if Pi?ol still enjoys trust and confidence of Duterte.
Lawmakers have also been calling for the resignation of NFA head Jason Aquino.
Pinol earlier proposed to have a rice trading center in Tawi-Tawi, citing the rampant rice smuggling in ZamBaSulTa area before.
The government's effort to curb rice smuggling in southern Philippines was one of the reasons why prices of rice went up, according to the DA chief earlier.
The National Food Authority unloaded in Zamboanga City over the weekend government-subsidized rice to lower the cost of the staple.
Zamboanga City previously declared state of calamity as prices of rice increased to as much as P70 per kilo.
The local government of Zamboanga City expressed hope that the price of rice would normalize soon with NFA rice in the market.
"We also understand that the amount of imports to be allowed will be enough to cover the needs of the ZambaSulTa area," Roque said.
"Again, we foresee the rice situation normalizing because the main harvest is beginning to come in by next month," he added. Celerina Monte/DMS


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Indicate hazard areas in your facility with Labelmaster's Warning "Caution" Tape. Available with a Caution message, a Caution/Do Not Enter message or yellow/black stripes with no message, this is made of vinyl with permanent adhesive to hold onto almost any clean, dry surface. The colors are sealed beneath a protective layer of abrasion-resistant, clear vinyl to stand up to your toughest indoor conditions. The striped aisle marking tape warns against potential hazards or caution against unsafe practices. Safety is your top priority in the workplace, and we go to great lengths to ensure our signs help you achieve your goals of awareness and compliance.
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This keychain features an Iron Rod with the words "Hold To The Rod" engraved into it. Perfect for Youth, Missionaries, and any member who wants a wonderful reminder of the Gospel.
Price: $12.99
Price: $3.99
Price: $3.55
Price: $2.49
Price: $2.95
Price: $3.49
Price: $17.99
I have had one of these key chains for years and I love it! I have given them to my kids and have recently purchased them to give out to my addiction recovery group and a dear friend who likes to have something tangible to remind them of Heavenly Father! I was so happy to see they are still available and a solid cool "iron rod" No other reviewer mr. Nerd it isn't really iron but I don't care. It's awesome!
This item, although a cool concept and I love having received it as a gift, does not contain any Iron as it is not magnetic. Stainless Steel would not tarnish, and would contain actual Iron, but this is not it. Hold to the Alloy Rod doesn't have the same ring to it to me, don't think any songs will be written about it...
My seminary teacher gave me one of these several years back. I bought three: gave one to my husband, a dear sister that has recently returned to Church, and gave one to a new friend on her baptism day. Just bought a few more to have on hand.
Love them, especially as goodbye presents for missionaries from our area that are returning home.
I purchased three of these so my wife, son, and I would each have one.
I'd had one when younger and lost it somewhere along my life, so I knew I wanted to get this as soon as ai saw it.
The key helps me remember to hold on to the iron Ron and gives me comfort.


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Winged Knits Candelia
Candelia is a top down, short sleeve cardi with raglan shaping, waist shaping and an optional peplum-ish detail at the back. The pockets are worked by continuing to knit some of the stitches from the fronts and then folded over and sew onto the body. Sample is shown with almost 2" of negative ease.
Finished Bust Measurements: 30.25 (32.25, 35.75, 39, 41.75, 45, 48.25, 51.75, 55)"
Yarn: approx. 700 (760, 865, 950, 1020, 1120, 1220, 1350, 1465) yards fingering weight yarn, shown in Siren Two Sock by Pigeonroof Studios (WEBS does not carry)
Needles: 16" circular and double pointed US 5; 29" circular US 6
Other Materials: stitch markers, large stitch holders, tapestry needle, buttons, hook and eye closures
Gauge: 24 sts and 32 rows = 4" in stockinette stitch using larger needles.


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GARLIC SELECT™ contains a unique blend of two highly effective, researched forms of garlic: garlic extract and black fermented garlic. These forms provide standardized levels of the researched organosulfur compounds allicin and S-allylcysteine, respectively.
Garlic has been used since ancient times to enhance immune resistance and support a range of health benefits. As a natural antibiotic, it offers broad spectrum antimicrobial activity and may be used to treat gastrointestinal issues such as dysbiosis and infections. Garlic also has been studied for its ability to help support healthy cardiovascular and immune system function, and to help support cholesterol levels already within the normal range.
High potency Garlic Select™ capsules are pleasant to take without any overbearing unpleasant garlic taste or odor. This product is rigorously tested for purity and potency and verified free of gluten by independent laboratory testing.
We aim to process and ship orders same day where possible. Please note all orders received after 3.30 pm on a Monday to Thursday will be processed and sent out the next working day.
Friday orders after 3.30pm will be shipped on the following Monday. For next day courier delivery orders must be received by 1.00pm.
For orders placed on Friday before 1.00pm requiring a next day courier you have 2 options; Next Day Saturday which incurs a premium charge or Next Day Standard which will be delivered on the following Monday.
Orders placed after 1.00pm via the courier option will be processed on Monday for Tuesday delivery.
Please note we do not process orders on Saturdays, Sundays or Bank Holidays.
|Supplement Facts
|Serving Size: 2 Capsules
|Servings Per Container: 30
|Amount Per Serving
|Garlic Extract (Bulb)(Allium sativum) (3:1 extract, 1% allicin)
|800 mg
|Black Fermented Garlic powder extract (yielding 1.2 mg S-allyl cysteine)
|22 mg
|†Daily Value not established.
Cellulose (capsule), microcrystalline cellulose, silicon dioxide, vegetable stearte.
Keep out of reach of children. Store sealed in a cool, dry place.
If taking medication, pregnant, or nursing, consult a physician before using.
2 capsules per day, preferably in divided doses, or as directed by your healthcare professional.


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Fun For Everyone!
Its over for another year.22-Oct-2018
Wow, what a great day even with some early drizzle!!
A fantastic day with plenty for everyone to enjoy. A huge thank you to our terrific community for their generous support.
Our heartfelt thanks to all our volunteers and stall operators for their commitment and effort. A great result which is only possible with your involvement.
Please continue to support our wonderful sponsors.
Next Fair:
Sunday 20th October 2019
8 am - 2 pm
At the grounds of Our Lady Star of the Sea Terrigal
165 Serpentine Road,
Terrigal NSW 2260


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Sign up for Meridian's Free Newsletter, please CLICK HERE
Cover image: Home by Greg Olsen.
Easter of 2018 is a remarkable and historic one for us as members of the Church. On Easter Sunday, (April 1, 2018) as we celebrate the risen Savior and His perfect life, we will all be together through the miracle of technology in General Conference. We will sustain President Russell M. Nelson as our Prophet, Seer and Revelator for the first time! We will also sustain a new apostle. We will gather in the knowledge and power that we are covenant members of the Lord's restored Gospel.
We need to be prepared for this event! I earnestly want to feel Him standing beside me and to expand my capacity to understand. I am prayerfully reviewing Easter scriptures and last October's conference. I am also treasuring two very special books that bring it all to life in marvelous ways that deserve to be shared.
A Shepherd Looks At Psalm 23
By Phillip Keller
Who doesn't love and carry vivid images in their hearts and minds of the 23rd Psalm? Who doesn't seek for ways to bring His perfect life and love more vividly into our minds and hearts with a wish for Him to heal our maladies great and small? Especially at Easter! Here two books that may bless you, as they have for me, with a greater sense of His being right beside me.
The gift of a couple of unexpected hours in a thrift store with shelves and shelves of books yielded an unexpected pearl of great price for me: A Shepherd Looks At Psalm 23 by Phillip Keller. A quick check on Amazon revealed how special it really is. First published in 1970, it is nearly 50 years old! It has been republished several times since, with many millions sold. There were nearly 900 joyful and tender-hearted reviews with comments that I now fully understand and agree to with equal gratitude.
Phillip Keller (1920-1997) was a shepherd in his youth and made it a full-time profession in his early adult years. He later became a pastor, a specialist in animal husbandry and a photographer who travelled the world. Unlike most of us in the 21st century who do not live on farms and know little of raising livestock, he knew the requirements for a flock of sheep to be healthy and well cared for. Just like King David who wrote the 23rd Psalm! As a pastor devoted to his own flock/congregation, he was inspired to create a series of sermons for his congregation based on his experiences as a shepherd. This book is the published version of those sermons.
In "A Shepherd Looks At Psalm 23", each line of the Psalm is its own chapter. From the first few paragraphs in the first chapter, 'The Lord is my shepherd, I shall not want,' it is surprising to see that there is far more sheer backbreaking work involved for the devoted shepherd than the idyllic pictures we have in our minds. The images of a serene and devoted shepherd with a crook, watching or holding a little lamb are a far cry from the long hours and days spent sowing seeds in pastures and getting up in the middle of the night to lead sheep to water. In these pages are born new visions of the great labors of the Savior and His devotion to us, new visions of ourselves as the needy and sometimes silly sheep who need such diligent tender care and new visions of what we can do to please Him.
While each line of the Psalm is its own chapter, unbeknownst to the average reader, King David is actually taking us through an entire year in the mindful tending of a flock. Brother Keller (the author of the book) does the same, each line of the poem and each season in turn, which is how David (the author of the Psalm) wrote it. Along with the flock's continual need for food, water, and protection from enemies, injury and illness, each season has its own particular challenges
For example In Chapter 8, "He maketh me to lie down in green pastures". It sounds so simple and serene! Yet, this is a very tricky feat for the shepherd because sheep (like young toddlers) have their own strong ideas about where and when to lay down. We learn that the purpose of lying down in the green pastures of Spring is to "ruminate" or chew their cud to properly eat and digest their food. The shepherd wants only to assist them, but they will not lie down if they are hungry, disturbed by the antics of other sheep, fearful of predators or bothered by flies, insects, the weather of the day or parasites. That hard-earned contentment of sheep lying down is a testament to a very skilled shepherd who has helped his sheep overcome those conditions and provide security for his flock!
There are many fascinating things to learn in each chapter, i.e. Chapter 8: "Thy rod and they staff, they comfort me." A shepherd's "rod" is not the long staff that we usually see. Instead, it is an additional strong stick that is chosen with great care from the sapling of a tree, then fashioned (using the knotty section of where the tree trunk branches into the roots) into a valuable and extremely personal tool that is considered to be a literal extension of the shepherd's arm. It is used in various ways to protect their sheep and flocks and to distract predators. Young shepherds may spend a lot of time choosing, making and learning how to use their rod. Some areas even have competitions and exhibitions where young shepherds throw their rods with great skill.
Sometimes it is used to lightly rest along the back of a favorite sheep while they walk along the trail as an expression of affection as the rod is literally an extension of the shepherd's arm.
The staff, sometimes with the crook as we are most familiar with, is a different tool used to gather and to herd the sheep.
After eloquently telling us about the shepherd's experience in caring for his flock with what each line is actually saying in terms of the care for a real flock of sheep, the author, who was also a pastor, then likens that segment to real life, real people, and the Savior's devotion and unending love for us, His little flock.
Perhaps the best way for you to feel the beauty and power of this book is to share a few paragraphs: Here are highlights of Chapter 10.
"Thou Annointest My Head With Oil …"
As one meditates on this magnificent poem, it is helpful to keep in mind that the poet is recounting the events in the full year in a sheep's life …. Here now, the sheep are in the summer.
Summer is a sublime setting on the high meadows, where there are clear running springs, where the forage is fresh and tender, where there is the intimate close contact with the shepherd. Suddenly we find "a fly in the ointment" so to speak.
In the terminology of the sheepman, "summer time is fly time." By this, reference is made to the hoards of insects that emerge with the advent of warm weather. Only those people who have kept livestock or studied wildlife habits are aware of the serious problems for animals presented by insects in the summer!
Sheep are especially troubled by the nose fly, or nasal fly, as it is sometimes called. These little flies buzz about the sheep's head, attempting to deposit their eggs on the damp, mucous membranes of the sheep's nose. If they re successful, the eggs will hatch in a few days to form small, slender, worm-like larvae. They work their way up the nasal passages into the sheep's head; they burrow into the flesh and there set up an intense irritation accompanied by severe inflammation.
For deliverance from this agonizing annoyance sheep will deliberately beat their heads against trees, rocks, posts or brush. They will rub them in the soil and thrash around against woody growth. In extreme cases of intense infestation, a sheep may even kill itself in a frenzied endeavor to regain respite from the aggravation. Often advanced stages of infection from these flies will lead to blindness.
Because of all of this, when the nose flies hover around the flock, some of the sheep become frantic with fear and panic in their attempt to escape their tormentors. They will stamp their feet and race from place to place. Some drop from sheer exhaustion. All this excitement has a devastating effect on the entire flock.
At the very first sign of flies among his flock, a good shepherd will apply an antidote to the heads of each sheep. I always used a homemade remedy made of linseed oil, sulphur and tar which as smeared over the sheep's nose and head a a protection against nasal flies … What an incredible transformation this would make among the sheep! This process was repeated individually for each sheep numerous times over the summer season.
This to me is the exact picture of irritations in my own life. How easy it is for there to be a "fly in the ointment" of even my most lofty spiritual experiences … not to mention daily life. So often it is the small, petty annoyances that ruin my repose ….
Just as the sheep needed a continuous and renewed application of oil to forestall the "flies", in my life there must be a continuous anointing of God's gracious Spirit to counteract the ever-present aggravation of personality conflicts. Only one application of oil, Sulphur and tar was not enough. It was a process that had to be repeated.
There are those who contend that in the Christian life one need only have a single, initial anointing of God's Spirit. Yet the frustrations of daily dilemmas demonstrate that one must have Him come continuously to the troubled mind and heart to counteract the attacks of daily small torments."
Just for fun, after reading the book, I went to Youtube.com and put in a search for videos of shepherds calling, herding and guarding their flocks. Not having ever spent time on a farm or with sheep, I confess, I cried in front of my computer when a shepherd from the Middle East called into a misty field for his flock with his rich accent. From the fog, soon many sheep were all running towards him! "They know my voice and I know them!" He said with love as he patted their heads. "Except for those," he said pointing off into distance with a little sigh and a chuckle. "Those two still have some learning to do!"
I chuckled as he did at these stubborn sheep who ignored him, but in truth it left me desiring more than ever to be one the Savior knows will come a-running the minute I hear His voice calling my name.
Each chapter/line is examined so carefully, thoroughly and eloquently that you will never look at a picture of the Savior with a lamb or a flock of sheep in the same way again. The author's perspectives of likening the sheep and their behaviors to human behaviors and needs in relation to the shepherd's skill and devotion has endeared the Savior to me in wonderful new ways that words cannot express.
Thank goodness that even though "A Shepherd Looks at Psalm 23" is nearly fifty years old, it is still in print to benefit each of us who wish to deepen our love, knowledge and understanding of the Savior and his role in our lives as the Good Shepherd.
While this book is not an LDS book, the other book I wish to share is.
The Healer's Art
Faith and The Healing Power of Jesus Christ
Lloyd D. Newell and Don H. Staheli
My mother, Barbara Brewerton Lee, was a full-time temple worker at the Washington, DC temple from 2001-2006. She had her own lovely apartment in the building the Church owns for the temple workers there. During those years she acquired many lovely gifts from her countless temple friends … especially books! She could not take them all with her when her time concluded. I was of the mind that she should leave them at the give-away table by the laundry room for others to enjoy. Not her! I can still see her with a stack of gift books, inscribing each one to someone she thought would need and appreciate it most. She was right. Those people were thrilled when we delivered them to have a little piece of my Mom.
The book that came my way – that I was so sure should have been left at the give-away table by the laundry room, has since become a treasure that I have read more than once and have given away as well to those I think might appreciate its healing power.
Like the 23rd Psalm that is so familiar, the piece of art that this book is based on is "Christ Healing at the Well of Bethesda" by Danish artist Carl Bloch and is equally dear to us. We've seen it countless times in Church publications and in our meetinghouses, but do we know much of its fascinating history? Or understand the significance of the lighting and the message behind each person conveyed in the picture?
The Healer's Art, just barely over 100 pages, is well worth the investment of a few dollars and a couple of hours to read it and learn more about the painting, the Savior and ourselves.
Few will know that it was some Lutheran priests that commissioned the painting in 1882 to celebrate the opening of a mission home. There it was treasured and revered for over 100 years as one of Denmark's most popular national works of art by the revered Danish painter, Carl Bloch.
In 2000 came the surprising query to BYU to see if they were interested in owning the painting! Why the mission home wanted to sell it and the dilemma of how the citizens of Denmark and the Danish government could say good-bye to their national treasure has the ups and downs, ins and outs and twists of plot of a riveting novel or movie. Its unveiling in 2001 at the BYU Museum of Art was a much celebrated event. It is now available to view at any time there in its own featured gallery. I have sat on the bench in front of it under the spell of the art itself, the Spirit and the message. I urge you to do the same if this opportunity comes your way.
"Wilt thou be made whole?" the Savior asks the man who for 38 years had laid by the pool with no one to help him be the first to enter the moving waters. (John 5:3-4)
The Savior, in a three-quarter length view slightly to the left of the center of the painting, is dressed in a full white robe. Tall and regal, the Savior gently extends his left arm to lift the covering over the man's tent-like covering. His right arm extends to the man, fingers pointing to this faithful man and his crippled limbs. The arc of the Savior's arms extending to uncover his face and then to reach out to him while pointing to his limbs is like a welcoming embrace that says, "I am personally here for you!" Though we barely see the man's face in the deep shadows of the tent-lilke covering, with his lame and bent legs extended in front of him, this represents each of us and the Savior's desire to lift the covering from our heads and eyes into light and hope of His love.
"I am personally here for you!" his arms symbolically say, pointing towards our covered heads and the problems we each have.
The significance of the light on the Savior's robe, the brightest part of the painting, is examined in the book, along with his magnificent role of physically healing individuals throughout time in every volume of scripture.
What is most interesting to me, and the call of the book, however is not the history of the painting or the history of the Savior's healing. It is the questions that are asked of each of us as we look at each of the other individuals in the painting. Do we come, like the mother in the far right edge of the painting with her water jug, child and a mother or mother-in-law beside her, every day for water, even living water and then share it with our families?
Perhaps even more poignant are the subliminal personal questions asked by the others in the painting. Their questions are summarized beautifully on the inside cover of the book:
"Are we like the distracted bystanders, too busy to notice the healing touch of the Savior? Or are we like the man in the red cap, waiting for a miracle but not fully ready to turn our pain over to the Master Healer? Perhaps we are among the crowd behind the pillar, separated by a barrier that stands between us and the Source of Salvation."
The book contains additional scripture stories, references, hymns and poems and is illustrated with
detailed sections of each individual in the painting. Tenderly written, it is a wonderful read aloud for youth and adults.
Of course the scriptures and the words of the living Prophets are our first source for feeling the Savior standing beside us. How blessed we are when gifted authors and artists are able to share their talents to enrich our knowledge and understanding even more to help us feel the Savior standing nigh, watching over our days, our problems, and our choices with His ever-present and loving desire to bring us safely home.
Carolyn Allen is the Author of 60 Seconds to Weight Loss Success, One Minute Inspirations to Change Your Thinking, Your Weight and Your Life. She has been providing mental and spiritual approaches for weight loss success both online and in the Washington, DC community since 1999 presenting for Weight Watchers, First Class, Fairfax County Adult Education and other community groups. She and her husband, Bob, are the parents of five children and grandparents of eleven. They are now happy empty nesters in Jackson, Tennessee, close to Memphis, where they center their online business for an amazing herbal detox drink/capsules.


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It's not easy to transition to a legal store, says owner of B.C.'s first approved private pot shop
Tamara Duggan says the province's legal licensing process is arduous, but rewarding.
Last week, Tamara Duggan's husband was pulled over by the local RCMP sergeant in their hometown of Kimberley, B.C. The town just received news that the Duggan's dispensary, Tamarak Cannabis Boutique, made history as the province's first privately owned pot shop to go legal. Naturally, red-and-blue in the rear-view would shoot an icy fear through the veins of anyone associated with the pre-regulated weed business, but the officer simply pulled him over to offer his congratulations.
"The news was surprising to everyone, but a really good surprise," she says on the phone to the Straight.
Duggan filed the application to legalize her dispensary on August 17, but says she has been preparing her customers for the transition since more serious talks of legalization began last year.
When she initially announced plans to move out of the grey market, however, not everyone was excited.
"When I started preparing my customers and making the transition arrangements there was a lot of panic," she says, adding that many of her regulars didn't know what to expect.
"We just encouraged them to stock up, as finances would allow, and we also provided some tips as to how to create your own edibles."
Edible and topical products didn't make the cut in the first round of legalized cannabis products, so Duggan and her team will be removing those items from their offerings. In the meantime, she has started educating customers on making their own infused cannabis products from their kitchens, adding legally purchased weed into recipes provided by the dispensary.
Duggan received the official confirmation that her application was approved last Wednesday (October 31). For three years prior, she operated the shop as a medical cannabis dispensary under a standard business licence.
"We're very well respected by our community, valued as good, corporate citizens and we're looked at as just another business," she says, adding her team has received nothing but support from the Kimberley crowd. She even credits the vote of confidence to the pace at which the shop received its seal of approval from the provincial government.
"They (the city council) posted in the newspaper twice, as well as passed out flyers to the surrounding community, asking for public feedback and consultation. The end result was 24 emails in response and none of them were of a negative nature," she says.
"That resounding support made it easy for the council to approve it and that sped my process of the application up for a fair bit."
Although Duggan says receiving the licence was a relief, the process came with a fair bit of hardship. She says it was clear the government was only a half-step ahead at every turn, and it was creating the new licensing program from scratch.
"I didn't realize what kind of trial-and-error process this was going to be," she says. Each time Duggan thought her checklist of documents and forms was complete, another would be added.
"It was when the inspectors come by the store that it was really confirmed to me just how close the applicant is to the development of the system," she says. Duggan explains once the final walk-through ended, an inspector directed her to an online portal to settle the licensing fees, only to discover the site hadn't yet been built.
"I called them and they said, 'yes, we're developing that page…today'," she says, laughing.
"It's kind of like I was the guinea pig in terms of how this process works, but it's been a rewarding process."
The whole process ran Duggan around $9,000—an application fee of $7,500 plus a yearly licensing fee of $1,500.
As it doles out private dispensary licences, the Liquor Distribution Branch—the authority charged with legal cannabis distribution in B.C.—is trying to weed out businesses with black market ties.
Along with the standard lot of operational information, applicants must disclose all personal banking details, including credit card debts, proof of assets, and income tax returns. They must also pass a several layers of criminal record, prior employment, and personal background checks.
"Whenever your life is splayed open it can be a bit of a stressful scenario," she says, reflecting on the process she calls "surprisingly invasive".
"You have to report everything, including a sealed juvenile record…they even wanted to know about charges you got but weren't convicted of."
Duggan says the breadth of information applicants must disclose is causing anxiety across the industry. Mmost businesses have operated in an unregulated and illicit market for years and much of what would be standard business practices are considered illegal activities. Admitting to those practices could come with consiquences.
Despite an arduous couple of months, Duggan says the provincial government has been "helpful and communicative" and is excited for the future of her business.
"There is lot's to look forward to. Instead of having multiple suppliers and struggling to source out vendors, going to one website with multiple vendors is going to be a dream," she says.
Although there will be a price increase, as government products are taxed at a higher rate, the store will now be able to offer smaller quantities of weed. Before legalization, Duggan says separating one gram baggies of dried flower was too time consuming. Now, she can just order pre-packaged quantities straight from the distribution centre.
"The smaller amounts gives customers a lot of opportunities for sampling before they commit to a strain they like," she says.
Duggan filed her first order with the LDB on Friday (November 2) and is optimistic the shop will be restocked and reopened by the end of this week.
"The nice thing is I am their only customer right now!" she says, laughing.
"Hopefully, any issues they run into they will be able to fix on the spot."More


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Weekly Tip: Siberian Ginseng
Siberian Ginseng has been around for thousands of years but only recently gaining popularity in the Western World. It also goes by the name Eleuthero.
The top 5 benefits of Siberian Ginseng
- Its an adaptogen - which helps your body adapt to various types of stress.
- Helps increase circulation of blood flow to the brain which improves concentration and memory.
- Enhances physical performance making it popular with athletes.
- Improves immune health and shortens the length of illnesses.
- Relives symptoms of fibromyalgia and chronic fatigue, relieving tiredness of the mind and body.
Related Posts
- Krystina's Weekly Tip Start your day off with a warm glass of water and table spoon of apple cider vinegar "the mother"
- Weekly Tip: Overnight Oats Quick and simple breakfast that you can prepare the night before.
- Homemade Basil Pesto It goes very well with a courgette dish I make which I will be posted during this month. Great if you are following a low carb diet.
- Weekly Tip: Propolis As the temperatures start to drop and we wave goodbye to summer this couldn't be a better time to start supplementing with Propolis.
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- Home
- apartment
- livorno
- san vincenzo
- Villa Sabbia Oro
Villa Sabbia Oro - apartment san vincenzo livorno
VILLA SABBIA D'ORO - An independent apartment immersed in the marvelous Tuscan countryside, set out on two floors, bright, spacious and elegantly furnished; at only 5 minutes from the sea and from the center of San Vincenzo.
It can cozily accommodate from 4 to 6 persons. The apartment consists of a living room, a dining room with a fireplace, 3 bedrooms, 2 bathrooms and a wide and equipped external area furnished with a table, chairs and a barbecue.
The guests of Villa Sabbia D'Oro may also take advantage of the services provided by the Sabbia D'Oro Hotel, among which the use of the wide beach, the restaurant and the breakfast buffet.
From the Villa it's very easy to reach Baratti with its marvelous beach, the crystal waters and the archaeological site, but it is also possible to reach some famous medieval villages such as Campiglia Marittima, Massa Marittima, Suvereto and also San Vincenzo, heart of the Etrurian Coast.
Furthermore, in an hour by car, it is possible to visit the main Tuscan cities such as Firenze, Siena and Pisa.
It is worth the time to visit the nearby natural park of Rimigliano, a protected environment perfect for mountain bike tours and trekking. There are many arboreal varieties such as juniper, cluster-Pine, stone pine and the rare maritime savine. The park is also populated by many different kinds of animals. You may often encounter the red squirrel. Harder to see, but certainly not impossible, the porcupine, the wild boar and the wild rabbit in the underbrush as well as the jay on the branches of the trees.
The thermae of Venturina, renowned since the Roman times, offer moments of pleasant relaxation, as well as the opportunity to receive specific treatments such as respiratory treatments, mud baths, massages and many others.
The wide external swimming pool, with thermal water at 34°C is entirely surrounded by parks. It is open during the summer period to allow people to relax by taking a nice dive or simply basking in the sun and having at ones disposal every comfort.


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Congratulations to DOGZPLOT BFF SHAINDEL BEERS.
from the publisher:
"Shaindel Beers to join Salt with a two book deal"
Fri, 1 Aug 2008 14:20:37 GMT
"Multi-award-winning poet Shaindel Beers is to join Salt with a two book deal. Beers' poetry, fiction, and creative nonfiction have appeared in numerous journals and anthologies. She is currently a professor of English at Blue Mountain Community College in Pendleton, Oregon, in Eastern Oregon's high desert and also serves as Poetry Editor of Contrary, and as a Poetry Reviewer for Bookslut."
Additionally, she took her Jeopardy test online and was chosen for an audition (I'm so jealous) and Alex told her how much smarter she was then everybody else and she promised me (no she didn't) if she don't know the answer to the final jeopardy question (she will) she will not guess, she will simply write "What is DOGZPLOT"


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If time is of a premium and you are realising that the beach body you set out to achieve as a new years resolution has not reached fruition…
or your wedding that you have spent years planning is only one month away and you have not yet got around to toning up for the big day then this is the package for you!
Our 4 week body blitz package is the ultimate in weight loss training and nutritional programmes!
Just fill out the online enquiry form or call to book your body blitz package!
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Saudi Arabia announced on Tuesday that it would now allow women to drive . Saudi Arabia is the only country in the world which does not allow women to drive. Women drivers have previously been arrested and cars have been confiscated. In a royal decree signed by king Salman bin Abdulaziz , the order said it will be effective immediately ... Read More »
Tag Archives: #SaudiArabiaFeed Subscription
A tweet to Sushma Swaraj brings Tripura man safely home from Saudi Arabia
A 32-year old man from Tripura was rescued by the Indian mission in Saudi Arabia after he complained about being ill treated by his employer in Saudi Arabia. Gopal Das, who went to Saudi Arabia nearly two years ago for a job, landed at Agartala Airport on Thursday. The resident of Barpathari village in South Tripura District claimed he had not ... Read More »
Saudi Prince announcement was a planned one?
The appointment of Mohammed Bin Salman as the next Saudi Crown Prince couple of months ago surpassing his uncle Mohammed Bin Nayef has started entering into controversies. As per the rule, Nayef is supposed to be the next Crown Prince after King Salman but the much younger Prince and the favourite son of King being appointed for the next in ... Read More »
Qatar-Saudi land border shut
Qatar's only land border connecting Saudi Arabia was shut on Monday leaving the otherwise bustling area looked deserted. Qatar which is scheduled to host the World Cup football in 2022 is in trouble after Saudi Arabia along with United Arab Emirates and few other gulf nations imposed sanctions on them for its alleged connection funding terror activities taking place from ... Read More »


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Price: 160 credits
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File Size: 129.52 Mb
Stacked MILF Works her DildoAmber Shyne | Directed by: Hekke1664 on July 5, 2016
This stacked MILF shows off her magnificent breasts, then spreads her legs wide, stroking and working a red dildo into her shaved pussy.
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June 30, 2018
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Single Idaho Gay Men Interested In Gay Dating
Looking for Idaho Gay Men? Look through the profile previews below and you may just see your ideal match. Send a message and setup a go out this week. Our site has thousands of other members waiting to date somebody just like you! Create your Totally Free Account Here
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I've had some problems in the past but I never let that get me down. I'm just an average guy who's looking for love. I'm looking for someone who will love mefor who I am and who isnt afraid of commitment or intimacy.


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Main Demo
Percussion Demo
March 5, 2019
Wav:274MB • 115 x WAv 44.1 kHz, 24-bit.
Welcome to Contemporary Tech House, a one-stop shop for modern sounding, fresh out of the box tech-house bangers!!
Taking influence from labels such as "Dirty Bird" and artists such as eats everything, Claude VonStroke, Green Velvet, Pleasurekraft… this pack is aimed to produce that top quality, fresh sounding tech house sound that will get heads turning & ears listening.
With sharp bass and groovy undertones, this pack delivers the goods.
The team here at Catalyst Samples are super excited to release this pack to the public after months and months perfecting this contemporary sound with 115 loops.
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*Other genres that this package may be useful in:
• House
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Inventory is not guaranteed. Please call location to verify availability.
Nike Lunar Fire Mens Golf Shoes. Your swings will be on fire with the Nike Lunar Fire Mens Golf Shoes. The ultralight Flywire technology will give you a custom fit while the Lunarlon cushioning provides responsive comfort. You'll feel stable and powerful as you swing with the Power Platform and flexible midfoot. Take on the course with confidence.
*Flywire technology *Lunarlon cushioning *Nike Power Platform *Seven removable CHAMP ScorpionSTINGER spikes *Stock# 853738


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History teaches us to respect the identity and expression of the fruits of the vineyard at each harvest.
Only with great sensitivity and deep knowledge can we transform the quality of the work undertaken in the vineyard into its expression as wine, and its balance into elegance, during its lifetime.
Its origins fuel the winemaking history of a region as fortunate as Ribera del Duero. History has brought us the Tinta del País, a local variety of the Tempranillo vine, a generous, exuberant vine, capable of achieving remarkable quality.
As every great tradition, ours must also reflect innovation; an updated vision can help to emphasise the wine's long history.


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Mini Review ARTICLE
Vascular Sap Proteomics: Providing Insight into Long-Distance Signaling during Stress
- Department of Biology, McMaster University, Hamilton, ON, Canada
The plant vascular system, composed of the xylem and phloem, is important for the transport of water, mineral nutrients, and photosynthate throughout the plant body. The vasculature is also the primary means by which developmental and stress signals move from one organ to another. Due to practical and technological limitations, proteomics analysis of xylem and phloem sap has been understudied in comparison to accessible sample types such as leaves and roots. However, recent advances in sample collection techniques and mass spectrometry technology are making it possible to comprehensively analyze vascular sap proteomes. In this mini-review, we discuss the emerging field of vascular sap proteomics, with a focus on recent comparative studies to identify vascular proteins that may play roles in long-distance signaling and other processes during stress responses in plants.
Plants are unable to relocate during unfavorable environmental conditions and instead rely on intricate signaling pathways that work at the local and systemic level to withstand stress. At the local level, stress-related signals move cell-to-cell through plasmodesmata (symplastic) or diffuse through the extracellular space (apoplastic). These communication processes are effective for signaling between neighboring cells or adjacent tissues, but are insufficient for systemic communication between organs (Lucas et al., 2013). For long-distance movement between distant tissues, macromolecules often access the plant vasculature; a system of specialized transport conduits connecting all organs of the plant.
The Plant Vascular System: Sharing Resources and Information
The xylem vessels and phloem sieve tubes of the plant vascular system connect above- and below-ground organs, allowing vascular plants to draw from two distinct resource pools. The xylem provides an avenue for the unidirectional transport of water and mineral nutrients (xylem sap) from roots to aerial tissues that is driven by the transpiration stream. In contrast, the phloem allows for the bidirectional movement of photosynthate and other macromolecules throughout the plant, from areas of synthesis or excess (source) to areas of use (sink) such as storage tissues or zones of active growth. The fluid within the phloem is known as phloem sap, and its movement is thought to be driven by a hydrostatic pressure gradient along sieve tubes according to the pressure-flow hypothesis (reviewed in De Schepper et al., 2013; Lucas et al., 2013). However, this idea is not fully supported as data demonstrating differential pressure between source and sink phloem has not been obtained (discussed in Knoblauch and Oparka, 2012).
In addition to its role in resource allocation, the vasculature serves as an important conduit for the exchange of information between organs. Phloem sap appears to be highly complex, containing a diverse set of molecules such as sugars, lipids, amino acids, peptides, proteins, coding and non-coding RNAs, small molecules, mineral nutrients, and phytohormones (Lough and Lucas, 2006; Turgeon and Wolf, 2009; Lucas et al., 2013). By comparison, xylem sap appears to be less complex, primarily containing mineral nutrients, peptides, proteins, and hormones (Lucas et al., 2013; Turnbull and Lopez-Cobollo, 2013). Many of these molecules have been implicated as signals or signal chaperones in a number of developmental and stress-related long-distance signaling pathways such as photoperiod-induced flowering, systemic acquired resistance (SAR), induced systemic resistance (ISR), wound responses, RNA silencing, autoregulation during plant-rhizobia symbioses, and systemic nutrient starvation responses. Most of these responses have been reviewed elsewhere (Lough and Lucas, 2006; Champigny and Cameron, 2009; Turgeon and Wolf, 2009; Lucas et al., 2013; Turnbull and Lopez-Cobollo, 2013; Lin et al., 2014), therefore we focus only on those that best illustrate how xylem and phloem facilitate inter-organ communication. The photoperiod-induced flowering response serves as the standard example for long-distance signaling in plants. Many plants sense increasing day length as a cue for the transition from vegetative to reproductive development. Early experiments demonstrated that a signal moves in the phloem from leaves, where day length is perceived, to the shoot apical meristem (SAM) where the transition to flowering is initiated (Zeevaart, 1976). More recent experiments identified the phloem-mobile signal as FT (FLOWERING LOCUS T) protein (Corbesier et al., 2007; Jaeger and Wigge, 2007). Interestingly, FT interacts with phospholipids that are important for FT's function in flowering-time regulation in the SAM (Nakamura et al., 2014). The lipid transfer protein DIR1 (DEFECTIVE IN INDUCED RESISTANCE 1) is an additional lipid-binding protein implicated in phloem-mediated long-distance signaling. During the induction of SAR, DIR1 and phloem-mobile immune signals move from locally infected to naïve distant leaves to protect against future pathogen infection (Champigny and Cameron, 2009; Champigny et al., 2013). Several hydrophobic small molecules (azelaic acid, glycerol-3-phosphate, dehydroabietinal, pipecolic acid) have been identified as potential SAR mobile signals in the phloem; however, further experimentation is required to understand how these molecules participate in SAR (discussed in Dempsey and Klessig, 2012). Xylem-mediated long-distance signaling has been implicated in certain abiotic stress responses. For example, nutrient deprivation induces xylem-mobile hormone signals (cytokinins and strigolactones) that travel from roots to shoots to alter plant development (Lucas et al., 2013). Long-distance signaling responses sometimes utilize both xylem and phloem for signal movement. Following infection with symbiotic rhizobacteria, plants produce long-distance signals that travel through the xylem from roots to shoots to inform autotrophic tissues of the impending symbiotic association. Upon the perception of this xylem-borne signal, a shoot-generated signal accesses the phloem and travels back to the roots to regulate the development of symbiotic structures (Staehelin et al., 2011). Xylem-borne CLE (CLAVATA3/EMBRYO SURROUNDING REGION-RELATED) peptide signals from roots are believed to interact with LRR–RLKs (leucine rich repeat–receptor like kinases) in shoots, which in turn induce the accumulation and movement of unidentified shoot-derived signal(s) back to roots (Searle et al., 2003; Staehelin et al., 2011; Wang et al., 2016). Together, these responses illustrate how the plant vasculature acts a conduit for information sharing between distant tissues.
Collecting Vascular Sap
Our current understanding of vasculature-mediated long-distance signaling is impacted by the challenge of obtaining pure vascular sap. Standard methods involve collecting the fluid that exudes from the cut ends of petioles or puncture wounds of stems. Phloem sap is easily collected from cucurbits and legumes, which exude large amounts of phloem sap from petiole cut ends and stems (discussed in Turgeon and Wolf, 2009). Notably, the purity of cucurbit exudates has recently come into question, as some members of this family exude primarily from extrafascicular phloem (non-transport), and xylem (Zhang et al., 2010, 2012; Zimmerman et al., 2013). In other plants, phloem sap is collected over the course of several hours by submerging petiole ends in EDTA-containing solutions, which prevents sieve element occlusion by limiting the availability of Ca2+ (King and Zeevaart, 1974). While EDTA-facilitated exudation enables phloem sap collection from a wide variety of plants, the sample is substantially diluted during the process and prolonged exposure to EDTA may lead to intracellular (non-phloem) contamination caused by tissue softening. A recent improvement to this method was described by Guelette et al. (2012), who demonstrated that an initial 1-h incubation of Arabidopsis petioles in EDTA, followed by a 9-h exudation period in sterile water, was sufficient for metabolomics and proteomics analysis. Another technique for the collection of phloem sap is aphid stylectomy, which uses phloem-feeding insects as tools to collect pure sap directly from phloem cells (discussed in Turgeon and Wolf, 2009). While the collected phloem sap is much less dilute, collection volumes are low and the method is technically challenging.
Xylem sap is typically sampled from the cut stems of larger plants such as Brassica oleracea, Zea mays, and Glycine max via bleeding or root pressure techniques. Bleeding techniques sample xylem sap directly from the cut end of stems or petioles, while root pressure techniques apply pressure (mechanically or through positive pressure using ice) to the rootstock to force liquid through the xylem, which is then collected from the cut end of the stem (Alexou and Peuke, 2013). Despite measures such as the pre-washing and blotting of cut stems, contamination of xylem sap by phloem and/or other cellular contents can be an issue for most xylem sap collection methods (Alexou and Peuke, 2013). Since small proteins and peptides have been implicated in long-distance signaling in the xylem (Neumann, 2007; Lucas et al., 2013), Okamoto et al. (2015) recently optimized a gel-free purification technique to enrich for small proteins/peptides in xylem sap. Combining o-chlorophenol extractions and HPLC (high performance liquid chromatography) separation, the authors identified small proteins and peptides that were not detected using electrophoresis based-methods (Okamoto et al., 2015). However, a major limitation of these methods is the inability to collect xylem sap from smaller plants, which are often used as molecular-genetic model systems.
Proteomics Analysis of Vascular Sap
Proteins play an important role in vasculature-mediated long-distance signaling responses, as demonstrated by the involvement of FT and DIR1 in photoperiod-induced flowering and SAR, respectively. Over the past 10 years, a number of proteomics studies provided information about the protein composition of xylem and phloem sap. Most studies relied on gel-based separation techniques such as 1D or 2D SDS-PAGE followed by standard protein detection approaches. This includes liquid chromatography (LC) coupled to mass spectrometers (MS), which consist of an ionization source (MALDI – matrix-assisted laser desorption/ionization, or ESI – electrospray ionization), and at least one of four types of mass analyzer; FTIC (fourier transform ion cyclotron), ion trap, TOF (time-of-flight), and quadrupole. Combinations of different mass analyzers in tandem MS set-ups improved protein coverage by overcoming particular weaknesses associated with each analyzer. In this review, we refer to these techniques simply as LC–MS/MS; for a comprehensive proteomics review, see Yates et al. (2009). Proteomics techniques have identified proteins in xylem and phloem sap collected from healthy plants growing in normal conditions (Table 1). In a recent review, Rodriguez-Celma et al. (2016) analyzed most of these studies and concluded that in general, the vascular fluids of multiple species contain proteins that appear to function in structural maintenance of the vasculature (e.g., cell wall metabolism) as well as constitutive defenses against pathogens (e.g., pathogenesis-related [PR] proteins, chitinases, proteases).
Comparative Proteomics Analysis of Vascular Sap
Comparative proteomics of vascular sap is an excellent approach to identify proteins that may be involved in long-distance signaling responses. Early comparative proteomics studies relied on 1- or 2D difference gel electrophoresis (DIGE) techniques to compare the protein profiles of different samples. Spots present in some samples but not others are excised from the protein gel and analyzed by LC–MS/MS. Unfortunately, this type of analysis performs poorly with more complex samples, since individual proteins cannot be resolved by electrophoresis. Moreover, DIGE techniques have limited coverage since only differentially abundant proteins are analyzed (Gemperline et al., 2016). Modern gel-free comparative techniques overcome both of these issues while also providing quantitative information for every protein that is identified. Gel-free approaches such as ICAT (isotope-coded affinity tag), ICPL (isotope-coded protein labeling), or iTRAQ (isobaric tags for relative and absolute quantitation) rely on the addition of chemical labels to protein samples. Alternatively, label-free comparative proteomics, in which separate MS runs are aligned and compared in silico, is used to identify differentially abundant proteins between samples/treatments. Label-free quantitation is much less labor intensive compared to label-dependent techniques such as iTRAQ, and often allows for superior protein detection and more accurate quantitation (Patel et al., 2009; Trinh et al., 2013). To date, each of these techniques has been successfully employed for comparative proteomics analysis of plant vascular sap during stress responses (Table 2). More detailed descriptions of these methods can be found in other reviews (Yates et al., 2009; Lottspeich and Kellermann, 2011).
Comparative Proteomics Analysis of Xylem Sap
Several comparative proteomics studies have been performed on xylem sap collected from plants experiencing stress (Table 2). The first was performed by Rep et al. (2002), who used MALDI-TOF-MS peptide fingerprinting to identify seven PR-related proteins that accumulate in xylem sap of tomato (Solanum lycopersicum) plants infected with the xylem-infecting fungal pathogen Fusarium oxysporum. This finding was later supported by a 2D-DIGE proteomics study that identified host PR proteins as well as pathogen-derived proteins in tomato xylem sap during F. oxysporum infection (Houterman et al., 2007). Recently, more comprehensive analyses of xylem sap collected during host interactions with F. oxysporum were performed using label-free quantitative proteomics in tomato and Brassica oleracea (Gawehns et al., 2015; Pu et al., 2016). Both studies identified substantially more total proteins (∼150–285) with a relatively high proportion of those proteins showing differential abundance during infection (Gawehns et al., 2015; Pu et al., 2016). Other xylem sap proteomes collected during plant–microbe interactions include Glycine max infected with Fusarium virguliforme (Abeysekara and Bhattacharyya, 2014), G. max during symbiosis with Bradyrhizobium japonicum or during treatment with elicitors from the pathogen Phytophthora sojae (Subramanian et al., 2009), and Brassica napus infected with Verticillium longisporum (Floerl et al., 2008). A common theme among these proteomes is the accumulation of PR proteins such as chitinases and glucanases, which may serve in an antimicrobial capacity to limit the spread of infection via xylem vessels (Sels et al., 2008).
Comparative proteomics analysis of xylem sap during abiotic stress has been studied in Zea mays, B. oleracea, and Gossypium hirsutum. Alvarez et al. (2008) analyzed xylem sap collected from well-watered and drought-stressed Z. mays using 2D-DIGE and LC-MS/MS, identifying 33 proteins that accumulated during drought and 8 that decreased in abundance. Fernandez-Garcia et al. (2011) analyzed xylem sap proteomes of salt-stressed and control B. oleracea plants using 2D-DIGE comparative proteomics, identifying 22 proteins that accumulated during salt stress and 18 proteins that decreased in abundance. More recent comparative proteomics analyses of xylem sap have focused on responses to nutrient limitation. Liao et al. (2012) analyzed the Z. mays xylem sap proteome during nitrogen-limiting and -oversupply conditions using 2-DIGE and Zhang et al. (2016) performed label-free quantitative proteomics on xylem sap collected from cotton (G. hirsutum) seedlings grown under normal and potassium (K)-limited conditions. Interestingly, the differential abundance of PR proteins, proteases, redox-associated proteins, and cell wall metabolism proteins was observed in each of these studies (Alvarez et al., 2008; Fernandez-Garcia et al., 2011; Zhang et al., 2016), which hints at the importance of these proteins in the xylem during abiotic as well as biotic stress.
Comparative Proteomics Analysis of Phloem Sap
Comparative proteomics analysis of phloem sap has been performed to investigate long-distance signaling during a number of stress responses (Table 2). Using iTRAQ-based proteomics Du et al. (2015) analyzed phloem sap collected from resistant and susceptible rice (Oryza sativa) cultivars that were either unexposed or exposed to phloem-feeding brown plant hopper (BPH) insects. They found that carbohydrate and protein metabolism proteins accumulated in phloem sap of susceptible BPH-infested plants, and that defense-related proteins accumulated in BPH-resistant plants. Responses to virus infection were investigated using phloem sap collected from melon plants (Cucumis melo) that were uninfected or infected with melon necrotic spot virus (MNSV). Using 2D-DIGE and LC-MS/MS, the authors identified a number of cell-death and redox-associated proteins that were differentially abundant during infection with MNSV (Serra-Soriano et al., 2015). Lastly, a label-free quantitative proteomics study was recently undertaken by our group to identify differentially abundant proteins in phloem sap of Arabidopsis thaliana during the induction of SAR with virulent or avirulent strains of Pseudomonas syringae (Carella et al., 2016). Of the 564 proteins identified in Arabidopsis phloem sap, 16 accumulated and 46 decreased in abundance during SAR. Proteins that accumulated in phloem sap during SAR included PR-1, redox-associated proteins, and putative lipid-binding proteins, while proteins with decreased abundance were associated with metabolism. The functional relevance of these proteins was investigated by performing SAR assays on corresponding T-DNA knockout mutants, which identified m-type thioredoxins (TRXm1 and TRXm4) and a major latex protein (MLP) as novel phloem-localized proteins that play functional roles in SAR.
Comparative proteomics analyses of phloem sap collected during abiotic stress responses have also been performed. An initial comparative study of phloem sap collected from hybrid poplar (healthy or mechanically wounded) identified 48 total proteins using 2D-DIGE and LC-MS/MS, with two proteins accumulating during wounding stress (Dafoe et al., 2009). A subsequent ICPL-based study of phloem sap collected from wounded and unwounded cucumber (Cucumis sativus) identified substantially more total and differentially abundant proteins (Gaupels et al., 2012). Interestingly, PR-type proteins were identified in the phloem sap of poplar and cucumber (Dafoe et al., 2009; Gaupels et al., 2012) and cucumber phloem also contained cyclophilins, carbon metabolism-related proteins, and other defense-related proteins (Gaupels et al., 2012). The phloem sap proteomes of plants experiencing other types of abiotic stress have been investigated in cucumber and B. napus. An iTRAQ-based study of phloem sap collected from salt-stressed cucumber identified several salt-responsive phloem proteins in salt-tolerant and -intolerant cultivars (Fan et al., 2015). In addition, comparative 2D-DIGE and LC–MS/MS analysis of phloem sap collected from Fe-deficient and control B. napus plants identified a number of redox-associated proteins with differential abundance during Fe stress (Gutierrez-Carbonell et al., 2015).
Common Proteins Present in Vascular Sap Proteomes during Stress
Comparative proteomics analysis of vascular sap has revealed a great deal about how the vascular system responds to stress. Surprisingly, a common theme among many xylem and phloem sap proteomes collected from stressed plants is the accumulation of PR proteins including thaumatin-like proteins, chitinases, glucanases, and MLPs. This may indicate that PR proteins play a role in the protection of the vasculature against pathogen/herbivore attack. Redox-related proteins (thioredoxins, peroxidases, etc.) are similarly associated with stress responses in the vasculature. It has been proposed that these proteins play a protective role in the phloem by preventing damage to proteins caused by oxidative stress (Walz et al., 2002). This may be especially important in phloem sieve elements as these cells lack protein synthesis machinery and cannot quickly replace proteins damaged during stress. In addition, cyclophilins, glycine-rich proteins (GRPs), and putative lipid-binding proteins are commonly identified in both xylem and phloem sap proteomes. The identification of multiple lipid-binding proteins in vascular sap supports the idea of lipid-based long-distance signaling in the vasculature (Benning et al., 2012; Barbaglia et al., 2016). The role of cyclophilins and GRPs in the vasculature is less clear. Extracellular GRPs have been linked to cell wall formation, while intracellular GRPs and cyclophilins have been implicated in a number of functions including RNA-binding/chaperoning, which may suggest a role in RNA-mediated long-distance signaling in the vasculature (Mangeon et al., 2010; Kumari et al., 2013; Rodriguez-Celma et al., 2016).
Conclusion and Future Directions
The emergence of sophisticated proteomics techniques has led to a new era in vascular sap proteomics studies. Together, these studies provide fundamental insights into the nature of plant vascular sap under normal and stress conditions. Although significant progress has been made in this field, much remains to be discovered. Further improvements to existing proteomics techniques will make it possible to detect low-abundance proteins essential for vasculature function. Moreover, novel xylem sap collection protocols for genetically tractable model systems are needed to investigate the importance of xylem-mobile proteins at the molecular-genetic level. In the meantime, researchers can take advantage of the Arabidopsis model system and the recently improved Arabidopsis phloem sap collection method (Guelette et al., 2012; Tetyuk et al., 2013) to expand our understanding of the molecular mechanisms of phloem-mediated long-distance signaling during plant stress.
Author Contributions
Conceived of the review: PC and RC. Wrote the review: PC, DW, and RC. Analyzed literature and created tables: PC and CK. All authors edited the manuscript.
This research was funded by a Natural Sciences and Engineering Research Council of Canada (NSERC) grant to RC, an NSERC graduate scholarship to DW, and an Ontario Graduate Scholarship to PC.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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Keywords: abiotic stress, biotic stress, long-distance signaling, phloem, proteomics, xylem
Citation: Carella P, Wilson DC, Kempthorne CJ and Cameron RK (2016) Vascular Sap Proteomics: Providing Insight into Long-Distance Signaling during Stress. Front. Plant Sci. 7:651. doi: 10.3389/fpls.2016.00651
Received: 23 March 2016; Accepted: 28 April 2016;
Published: 12 May 2016.
Edited by:Hanjo A. Hellmann, Washington State University, USA
Reviewed by:Abu Hena Mostafa Kamal, University of Texas at Arlington, USA
Elisabeth Jamet, Laboratoire de Recherche en Sciences Végétales, France
Copyright © 2016 Carella, Wilson, Kempthorne and Cameron. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Robin K. Cameron, firstname.lastname@example.com


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3(1)Civil Procedure Rules may provide for the removal of proceedings at any stage—
(a)within the High Court (for example, between different divisions or different district registries), or
(b)between county courts.
(2)In sub-paragraph (1)—
(a)"provide for the removal of proceedings" means—
(i)provide for transfer of proceedings, or
(ii)provide for any jurisdiction in any proceedings to be exercised (whether concurrently or not) elsewhere within the High Court or, as the case may be, by another county court without the proceedings being transferred, and
(b)"proceedings" includes any part of proceedings.
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