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<urn:uuid:9b30419b-e4cb-4a00-9b83-56c7572fb185>	FlightSafety International is pleased to announce that Timothy Decoursey has been promoted to Assistant Manager of its Learning Center in Columbus, Ohio. “We are fortunate to have Tim as a member of our team in Columbus, and are pleased that he has joined our management staff,” said Chip White, Manager, Columbus Learning Center. “Tim’s management experience, leadership abilities, and dedication to provide outstanding service and the highest quality training is valued and appreciated by our Customers and Teammates.” He joined FlightSafety at the Columbus Center in 2002 as a Cessna Citation Ultra instructor. He has held a number of positions of increasing responsibility since then including Assistant Program Manager for the Citation X and Program Manager for the Citation Ultra. Tim replaces Fabio Miguez who was recently promoted to Manager of FlightSafety’s Learning Center in Toledo, Ohio. Decoursey was with American Airlines prior to joining FlightSafety International. He served as a Flight Engineer on the Boeing 727 and as First Officer on the McDonnell Douglas MD-80 and DC-10 aircraft. Tim earned a Bachelor of Science Degree in Education from the University of Illinois and a Master of Business Administration from Northern Illinois University. He holds an Airline Transport Pilot Certificate and Type Ratings in the Cessna Citation CE-500 and CE-750 aircraft. He also holds a Flight Engineer Turbojet certificate and is a licensed Powerplant mechanic. FlightSafety’s Columbus Learning Center offers a wide variety of training programs for aircraft manufactured by Dassault Falcon, Cessna Aircraft Company, Hawker Beechcraft Corporation and Gulfstream Aerospace. The Center features eight FlightSafety designed and manufactured full flight simulators and other advanced training devices and equipment that are part of MATRIX, FlightSafety’s integrated training system. FlightSafety International is the world’s premier professional aviation training company and supplier of flight simulators, visual systems and displays to commercial, government and military organizations. The company provides more than a million hours of training each year to pilots, technicians and other aviation professionals from 154 countries and independent territories. FlightSafety operates the world’s largest fleet of advanced full flight simulators at Learning Centers and training locations in Asia, Europe, North and South America, the Pacific Rim and South Africa.
<urn:uuid:67cca9d8-d7f4-4125-babe-34736e5a60aa>	Arts & Culture A Jewcy-Approved Coachella Lineup In some music circles, you’re not cool unless you’ve sweated it out with the thousands of in-the-know music snobs and the people who love them at the hipster music fest Coachella which takes place in Indio, California. Besides the hundred-plus … Read More In some music circles, you’re not cool unless you’ve sweated it out with the thousands of in-the-know music snobs and the people who love them at the hipster music fest Coachella which takes place in Indio, California. Besides the hundred-plus degree temperatures and the endless port-o-potty lines, Coachella typically boasts a covetable lineup of the latest and greatest of the sonic scene. This year, we at Jewcy are happy to report that there is a generous smattering of Jewish artists scheduled to grace the scorching stage. Iconic songwriter, and poster child for the perennially morose, Leonard Cohen is slated to round out the main acts on Friday night’s roster. While not the most upbeat, Cohen definitely meshes with the comeback cool theme of Coachella (Paul McCartney, anyone?) Another mentionable Jewess taking the stage is the notorious Amy Winehouse, whose super-sized beehive and solid pipes have been recently overshadowed by her crack habit and good-for-nothing convict husband. Whether the trouble-making queen of British soul with even make it to the desert is a whole other question—how can she be eligible for a Visa?—but after seeing her play a sweat-inducing set in a tiny club in San Francisco, I’d be willing to wait in 103 degree desert heat just to hear "Rehab" live. Check out the full lineup here, while I quietly contemplate what Morrissey’s famous pomp will look like in that weather.
<urn:uuid:ff4eacc6-7e16-433b-b0e6-c6dd9a4fd499>	In response to the June 9 post about memories of the CIA in Miami, JFK Facts contributor Arnaldo Fernandez sent this photo of a group of Cuban frogmen shortly before the invasion at the Bay of Pigs. “They started training on the Miami River aboard the WW II assault landing craft Blagar, which would be the command ship in the invasion. Their final trainer was CIA paramilitary operations officer Grayston Lynch,” Fernandez writes. “According to Bradley Ayers, a U.S. Army officer assigned to JM/WAVE for undercover operations,” Fernandez goes on, “Grayston Lynch was one of the nine people based at the CIA Station in Miami, who would ‘have intimate operational knowledge of the circumstances surrounding the [Kennedy] assassination.’ The other eight, according to Ayres, were station chief Ted Shackley, David Sanchez Morales, Rip Robertson, Edward Roderick, Tony Sforza, Thomas Clines, Gordon Campbell, and Felix Rodriguez.” I can’t accept Ayres’ assertion. Yes, the CIA’ still-secret records about David Morales are among the most important JFK records that remain subject to government censorship. Yes, Tony Sforza, known inside the CIA as “Henry Sloman,” was an assassin, according to Seymour Hersh. But Gordon Campbell, who ran maritime operations for the CIA, died in September 1962. So the claim that he had “intimate operational knowledge” of JFK’s assassination 14 months later is unfounded or supernatural. Here’s Campbell’s death certificate.
<urn:uuid:58ce2109-98a4-428f-b82e-5a5d27b40fba>	You are here: 20th Anniversary Tribute To Jim Carrey27 Oct 2016 By Eva Araújo (Web correspondent) If you're a Jim Carrey fan, this website is for you but if you're a fan who has something to say, this article in particular will meet your needs. In the year that we celebrate the 20th anniversary of this website and of our Jim Carrey Online community we decided to do something a bit different to give a voice to all Jim's fans out there and all over the world! And for that we need you! We need you to simply let us know what you think about Jim Carrey! You can write a text, a message, a letter, a poem, whatever your heart desires and all you feel you would like Jim to know. This is a tribute celebration to the man who brought us all together and to whom we love and care about. Every little word count and it will be featured in our social media accounts and also this website. And more important than that… it will reach Jim's heart! That's the goal! So what are you waiting for? Send us your opinion to this email: [email protected] It doesn't matter what language you use, or where you are from. All fans are welcome and all are necessary! Let's show Jim Carrey how much we love him! We at JCO will thank everyone who participates with a promise of continuing to keep you updated as more news come. -- Click to comment this article. » Send to friend « Newer article \| Overview \| Older article »
<urn:uuid:117ed7fb-e783-4448-bb4d-7b664c1d80d7>	After, we finally got to see Robb & Lewis's new condo that they are preparing to renovate. I can't wait to see my new summer home in Vancouver when it's completed. We did a very speedy tour of Winner's (the Marshals of the North) and then it was time for Robb's mum to go home and me to get a nap. The funny part of the Winner's tour was it was Robb & I who were aching to get our shop on and his mum only came along reluctantly. Fast forward to 45 minutes later and mum is the only one making a purchase as Robb and I droop there empty handed. Instead Lyle and I got coffee downstairs from the condo at the shop across Davie street called Coo Coo Cafe. C'mon, there is coffee on every corner! Literally, EVERY CORNER. IF we had wanted to, there was another coffee shop downstairs across the street to the West, one of a chain called Blenz. There were also two Starbucks within two blocks to the East, one after another, in the same direction.) Saturday night we went to a friend's house back in Kitsilano and they served dinner on the rooftop deck. It looked like an ad for gracious living in a gay magazine. "Say yes to expensive champagne, you've earned it with your attractive friends," type of ad. After dinner just the four of us walked to the water (uh, half a block, those dinner friends have a great location) to better view the fireworks over English Bay. The Symphony of Fire lasts for two weeks. The first three nights of competition are solo competitors. The final night is a selection of all three then a grand finale presented by the sponsor. We were there for the finale. Most people go to the beach early and stake out their prime viewing spot starting around 4 or 5 in the evening for a 10 pm show. We walked down at 9:50, leaned against a fence around a pool until 9:59, then moved forward to the back of the crowd at 10. We were 6 rows from the waterfront. Someone else had brought a radio so we scooted closer to be able to hear the music that went with each performance. I think we can all agree that firework photos are boring after the show. But since this is my blog, I get to post them anyway. These are my greatest hits. Click on the photo above to see it bigger. Fireworks take my breath away. My favorite part is the BOOM that pushes your chest in. In my book, the closer to the launch zone the better so you can really feel the boom. After fireworks, we walked all the way back to the West End of Vancouver. About a 2 miles. I have a theory that I normally adhere to, outdoors is a place you pass through when you get out of your car at valet and have to get into the mall. Walking is what you do between stores. Was this my life this weekend? We did not go out Saturday night. (close your mouth, it happens.)
<urn:uuid:a91db8fd-3959-4f42-a751-63b80671da77>	I've fallen behind on my picture book reviews of late. So I've decided to try something new. I'm going to do small round-up posts featuring my favorites of the titles that I've received from various publishers. First up was Kane Miller. Second was Eerdmans Books for Young Readers. Third was Chronicle Kids. Today I am featuring five titles (or so) from HarperCollins Children's Books. 1. Paddington in the Garden, written by Michael Bond and illustrated by R. W. Alley. In light of the recent movie, HarperCollins has been issuing and re-issuing various editions of Paddington stories. Among my favorites are the picture books, including Paddington in the Garden. This story, lovingly illustrated by R. W. Alley, focuses on Paddington's love of the Browns' garden, and his joy when he's given a little patch of garden to maintain on his own. A contest ensues, and mishaps occur, but in the end, Paddington triumphs. Paddington in the Garden is text-heavy as picture books go, reflecting Paddington's original status as a chapter book character. There are sentences like: "They (gardens) usually require a lot of hard work, and the one at number thirty-two Windsor Gardens was no exception." Parents reading this aloud may have to stop and explain some things. However, the Paddington picture books still provide an accessible entry point for kids into the world of the Paddington stories. Alley's busy, colorful illustrations will make readers of all ages smile. My own daughter gave the first Paddington chapter book a try, but (at five) she likes the picture books better. 2. Paddington at the Beach, written by Michael Bond and illustrated by R. W. Alley. Paddington at the Beach is a shorter, simpler story than Paddington in the Garden. It focuses on a visit that Paddington makes, by himself, to the beach, and his interactions with a pack of greedy seagulls. Paddington at the Beach is also a counting book, as seagulls 1 to 10 arrive gradually over the course of the story. This book feels more like it was conceived as an original picture book (vs. an adapted incident from one of the chapter books), though I don't know this for certain. Regardless, there is still a relatively sophisticated vocabulary that will reward parents and children reading together. Other Paddington titles that we've been reading include the original Paddington picture book and a The Paddington Treasury: Six Classic Bedtime Stories. I've also seen my daughter looking through a couple of movie tie-in edition paperbacks, but she hasn't asked me to read those to her. I suspect we'll get back to the chapter books soon... 3. Goose, by Laura Wall. Goose is the story of a lonely girl named Sophie who wishes that she had someone to play with. At the park, she befriends a goose. They play together on the seesaw and the slide and have a great time. Eventually (after nearly going off for the winter with the other geese), Goose is allowed to go home with Sophie. I like that Goose is just ridiculous, with no moral messages or larger theme (beyond the idea that it's nice to have someone to play with). There's a boy playing in the park with his teddy, and I expected Sophie to end up playing with him, but no, Wall avoids this obvious solution in favor of Goose as playmate. There's an old-fashioned feel to Goose (Sophie and her mom both wear dresses, and there's no question of her mom playing with her in the park), but this is balanced by bright, eye-catching illustrations. The illustrations were drawn in charcoal, and then digitally colored. Each page has a single-toned, bright background. The text is brief and straightforward, meaning that Goose could also work well as an early reader. But I think that Goose will work best as a lap read-aloud for very young readers, who can practice identifying the colors, and laugh at the idea of a Goose on a slide. 4. Goodnight Already!, written by Jory John and illustrated by Benji Davies. Goodnight Already! is the story of Bear, who just wants to sleep, and his neighbor Duck, who is awake and looking for company. Duck keeps hounding Bear, and Bear responds with ever-increasing levels of grouchiness. Until, eventually, Duck falls asleep, and Bear is left wide-awake and frustrated. There's a long tradition of books like this, of course, with a pesky friend trying to win over someone grouchy (who often is a bear). What I like here is that Bear never is won over (even though he is woken up). No matter what Ducks asks ("Want to play cards?" "Can I borrow some sugar?") the answer is always "No." There's also plenty of humor, both in the dialog and in the illustrations. My favorite scene is one in which Duck wakes up Bear by poking him on the nose. Bear's eyes pop open, and so do the eyes of Bear's pink stuffed bunny. I laughed out loud. I'm actually surprised that this book has a different author and illustrator, because the text and illustrations mesh perfectly. I recommend Goodnight Already! for home use, because it's a perfect parent-child read-aloud. The only risk in reading it as a bedtime book, of course, is that the child will be kept awake from laughing. But that's a small price to pay. Goodnight Already! is going on my keep shelf. 5. Mahalia Jackson: Walking with Kings and Queens, written by Nina Nolan and illustrated by John Holyfield. Very different in tone from the picture books profiled above, Mahalia Jackson: Walking with Kings and Queens is a picture book biography about a girl from a poor family in New Orleans who grew up to be a famous gospel singer. Nolan's prose somehow begs to be read with a Southern accent. When her aunt tells her "One day you'll walk with kings and queens", what follows is: "How was Mahalia going to walk with kings and queens? She didn't even have shoes. But Aunt Bell had a way of knowing things." Her aunt is strict, and takes her out of school after fourth grade, but she still tells Mahalia to believe in her own greatness. That aunt is remarkable, as is Mahalia's story. There's a lack of melodrama to Nolan's recounting. Just the facts. Out of school, back in school, working hard, but always singing. Holyfield's acrylic illustrations add more softness to the book. They are works of art, too, capturing the various settings experienced in Mahalia's journey, as well as the affection between Mahalia and her relatives. Young readers will, I think, be inspired by Mahalia's dedication to her music, and to her religion. If she could rise up from nothing and sing in front of prime ministers and presidents, it will be hard for readers to complain. I hope that kids will find Mahalia's story. Recommended for school and public library purchase. © 2015 by Jennifer Robinson of Jen Robinson's Book Page. All rights reserved. You can also follow me @JensBookPage or at my Growing Bookworms page on Facebook. This site is an Amazon affiliate, and purchases made through affiliate links (including linked book covers) may result in my receiving a small commission (at no additional cost to you).
<urn:uuid:4d6bb1a8-a384-45e6-a574-7939e84f591e>	Juanita Jean, pretty much sums up how many Americans view terrorism. When a muslim commits an act of terrorism it is viewed as part of a muslim conspiracy. But when a christian commits an act of terrorism it is viewed as the act of a loner and nut-job. Personally, I don't see the difference. Neither is acting completely alone with no input from anyone else, but to consider either one as representative of their stated religion is also wrong. Terrorists are all vicious criminals regardless of their religion (or lack of it) and no one is born to be a terrorist. It is a behavior that is learned from experiences and from listening to others (and those others may not even realize they are helping to create a terrorist). When we demonize those who we disagree with and talk as though they are less than human, then we may well be creating a future terrorist. The Norwegian terrorist was not a lone nut. He learned his beliefs and justified his behavior by reading and listening to others -- just as all terrorists do.
<urn:uuid:ffe63bd9-0aba-4163-bfa5-c580e7dbd96b>	I write a lot about radically altering how we interact with children. There is a big difference between working with children and doing things to them. You could choke a horse with the sheer number of reward and punishment based programs that have infected our schools and homes. Some teachers and parents believe in using rewards or punishments in an attempt to encourage initial interest in something. While these videos are far from definitive evidence for abolishing behvaiour programs, I think you can appreciate how the manipulated feels toward to the manipulator. These kids wear their feelings of disgust and disapproval on their faces in plain view. But how many times do kids walk away from interactions with manipulating parents and teachers and never show their true feelings for us? We can try and tinker with these bait and switch programs; we could fill conferences discussing whether we start with the big red toy truck and wean kids off to the cardboard cut-out or start with the plastic horse and graduate to the real one - but this all misses the point. I'm not prepared to gamble with the relationships I have with my students or daughter to gain some kind of compliance.
<urn:uuid:e1b4ca82-84f4-4be5-921e-aea05fc9468e>	Boycott Jamaica Site Launched Activists/bloggers Wayne Besen, Jim Burroway, and Michael Petrelis have jointly launched a site calling for the boycott of Jamaica, "the most homophobic place on Earth." Human rights activists have given Jamaica the infamous title: “The Most Homophobic Place on Earth.” If you love your gay friends and family members, you won’t visit Jamaica. If you care about the human rights of gay, lesbian, bisexual and transgender people, you won’t buy Jamaican products. Isn’t it time we stop rewarding this hate state with our tourism dollars? Isn’t it time to stop drinking Jamaican beverages, such as Myers Rum and Red Stripe Beer?Go here to sign up for updates from the site. This nation should be avoided at all costs until the Jamaican government takes action to end the country’s virulently homophobic climate and draconian laws that persecute homosexuals. Until Jamaica takes the following easy steps an official boycott is in effect: 1) Publicly commit to end gay bashing on the island and improve the human rights situation 2) A statement from the Prime Minister clearly and unequivocally condemning violence against GLBT people and expressing regret for past violence
<urn:uuid:d0f21953-53cd-42e4-9ebf-65139990db8b>	Infighting Is Funny, Part 96 "Ann Coulter has lied about and vilified conservatives who threaten the candidacy of Mitt Romney, whom she calls the “most conservative” candidate. Just this week, she again falsely claimed Romney to be the most conservative, falsely asserted that Sarah Palin is an establishment Republican, and falsely contended that Romney really won in Iowa. Coulter has also dismissed the Tea Party as conspiracy nuts to promote Romney. Coulter gushes over Romney and even defended Romney Care, now known as Coulter Care! Wanting so desperately to be a king-maker, Coulter is resolved to foist her choice upon the rest of the nation, using treachery and deceit to achieve her goals. If she will lie about conservatives (even though she is considered a conservative leader and icon), why wouldn’t she lie about liberals (whom she claims to hate)?" - Daniel Borchers, founder of Coulter Watch.
<urn:uuid:9c41d42d-3be9-4a63-9434-78af96858f6c>	Objective: Electroconvulsive therapy (ECT) is one of the most effective treatments for severe major depressive disorder. However, after acute-phase treatment and initial remission, relapse rates are significant. Strategies to prolong remission include continuation phase ECT, pharmacotherapy, psychotherapy, or their combinations. This systematic review synthesizes extant data regarding the combined use of psychotherapy with ECT for the treatment of patients with severe major depressive disorder and offers the hypothesis that augmenting ECT with depression-specific psychotherapy represents a promising strategy for future investigation. Methods: The authors performed 2 independent searches in PsychInfo (1806-2009) and MEDLINE (1948-2009) using combinations of the following search terms: Electroconvulsive Therapy (including ECT, ECT therapy, electroshock therapy, EST, and shock therapy) and Psychotherapy (including cognitive behavioral, interpersonal, group, psychodynamic, psychoanalytic, individual, eclectic, and supportive). We included in this review a total of 6 articles (English language) that mentioned ECT and psychotherapy in the abstract and provided a case report, series, or clinical trial. We examined the articles for data related to ECT and psychotherapy treatment characteristics, cohort characteristics, and therapeutic outcome. Results: Although research over the past 7 decades documenting the combined use of ECT and psychotherapy is limited, the available evidence suggests that testing this combination has promise and may confer additional, positive functional outcomes. Conclusions: Significant methodological variability in ECT and psychotherapy procedures, heterogeneous patient cohorts, and inconsistent outcome measures prevent strong conclusions; however, existing research supports the need for future investigations of combined ECT and psychotherapy in well-designed, controlled clinical studies. Depression-specific psychotherapy approaches may need special adaptations in view of the cognitive effects of ECT. From the *Department of Psychiatry, The University of Texas Southwestern Medical Center, Dallas, TX; and †Division of Brain Stimulation and Therapeutic Modulation, Department of Psychiatry, New York State Psychiatric Institute, Columbia University, New York, NY. Received for publication March 10, 2010; accepted August 30, 2010. Reprints: Shawn M. McClintock, PhD, The University of Texas Southwestern Medical Center at Dallas, Department of Psychiatry, 5323 Harry Hines Blvd, Dallas, TX 75390-8898 (e-mail: [email protected]). This publication was supported by Grant Number KL2RR024983 (Principal investigator [PI]: Milton Packer) from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH); Grant Number K23MH085007 (PI: A. Brandon) and Grant Number K24 MH-001571 (PI: R. B. Jarrett) from the National Institute of Mental Health (NIMH); and Grant Number 05T-682 (PI: M. M. Husain) from the Stanley Medical Research Institute. Shawn M. McClintock PhD and Anna R. Brandon PhD contributed equally to this article.
<urn:uuid:87f21ae1-01ea-4e94-b6d8-c67c01f4442c>	During pregnancy, syncytiotrophoblast vesicles contribute to maternal tolerance towards the fetus, but also to pathologies such as pre-eclampsia. The aim of the study was to address whether Plasmodium falciparum and HIV infections in pregnancy affect the secretion, microRNA content and function of trophoblast microparticles. Microparticles were isolated and characterized from 122 peripheral plasmas of Mozambican pregnant women, malaria- and/or HIV-infected and non-infected. Expression of placenta-related microRNAs in microparticles was analysed by qPCR and the effect of circulating microparticles on dendritic cells assessed by phenotype analysis and cytokine/chemokine measurement. Concentrations of total and trophoblast microparticles detected by flow cytometry were higher in HIV-positive (P = 0.005 and P = 0.030, respectively) compared to non-infected mothers, as well as in women delivering low birthweight newborns (P = 0.032 and P = 0.021, respectively). miR-517c was overexpressed in mothers with placental malaria (P = 0.034), compared to non-infected. Microparticles from HIV-positive induced a higher expression of MHCII (P = 0.021) and lower production of MCP1 (P = 0.008) than microparticles from non-infected women. In summary, alterations in total and trophoblast microparticles associated with malaria and HIV in pregnant women may have an immunopathogenic role. The potential for placental-derived vesicles and microRNAs as biomarkers of adverse outcomes during pregnancy and malaria infection should be confirmed in future studies. Citation: Moro L, Bardají A, Macete E, Barrios D, Morales-Prieto DM, España C, et al. (2016) Placental Microparticles and MicroRNAs in Pregnant Women with Plasmodium falciparum or HIV Infection. PLoS ONE 11(1): e0146361. doi:10.1371/journal.pone.0146361 Editor: Adrian J.F. Luty, Institut de Recherche pour le Développement, FRANCE Received: May 25, 2015; Accepted: December 16, 2015; Published: January 12, 2016 Copyright: © 2016 Moro et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability: All relevant data are within the paper and its Supporting Information files. Funding: This work was supported by the Instituto de Salud Carlos III (PI13/01478, CES10/021-I3SNS to AM); the Banco de Bilbao, Vizcaya, Argentaria Foundation (BBVA02-0); the Ministerio de Ciencia e Innovación (RYC-2008-02631 to CD); the Institut d'Investigacions Biomèdiques August Pi i Sunyer (to LM) and the Departament d’Universitats i Recerca de la Generalitat de Catalunya [Agencia de Gestión de Ayudas Universitarias y de Investigación (AGAUR); grant 2014SGR263]. The Manhiça Health Research Centre receives core support from the Spanish Agency for International Cooperation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing interests: The authors have declared that no competing interests exist. Infectious pathogens can alter the release and function of extracellular vesicles (EVs) to promote growth and induce transmission, evade host immune system and manipulate cellular microenvironment [1,2]. Exosomes, 30–100 nm and originated from inward budding of endosomal membranes, and microparticles (MPs), 100–1000 nm and originated from outward budding of plasma membrane , participate in intercellular communication, including immune functions, in physiological and pathological conditions [1,4]. Key players in functions mediated by EVs are secreted microRNAs (miRNAs), small non-coding RNAs that trigger either mRNA degradation or translational repression . These EVs and miRNAs can be measured in body fluids and present a potential as diagnostic biomarkers [6,7], although standardized procedures are lacking . Little is known about the role of EVs in malaria and human immunodeficiency virus (HIV) infections, two of the major health priorities especially for pregnant women and their infants. Circulating plasma and red blood cell-derived MPs levels increase with malaria severity , especially in cerebral malaria , and lead to the activation of brain endothelium [11,12]. Genetically deficient mice with impaired capacity to vesiculate are completely resistant to cerebral malaria , thus confirming the implication of EVs in the physiopathology of the disease. Importantly, exosome-like vesicles seem to be implicated in cell-to-cell communication between Plasmodium-infected red blood cells for the initiation of parasite sexual differentiation , and exosomes from Plasmodium yoelii-infected reticulocytes elicit protective immune responses against lethal infection in mice . EVs can also contribute to viral infection and pathogenesis , such as in the case of HIV, by facilitating macrophage infection , inhibiting antigen-presenting cells responsible for early pathogen detection (i.e., dendritic cells (DCs) ), and transferring the HIV co-receptor chemokine receptor 5 (CCR5) to other cells . EVs also have an important role in the communication between fetus and maternal immune cells necessary for pregnancy success . Syncytiotrophoblast-derived MPs contribute to maternal tolerance towards the fetus [21,22], but can lead to pre-eclampsia-associated inflammation . Trophoblastic MPs (trMPs) contain placental miRNAs, also present in maternal plasma and breast milk , trophoblast cells from placenta or cultured lines , and exosomes from villous trophoblasts . Placenta-derived miRNAs [24,27] participate in fine-tuning of gene expression in the fetal-maternal interface and are altered in pregnancy-related pathologies such as pre-eclampsia, intrauterine growth restriction (IUGR) or preterm delivery . However, no previous study has investigated the relationship of trophoblast-derived vesicles with malaria and HIV infections in pregnant women. We hypothesized that modifications in the placenta caused by Plasmodium falciparum and HIV [30,31] affect trMP secretion contributing to altered pregnancy immune responses. Therefore, our primary objective was to study ex-vivo the effect of P. falciparum and HIV infections on trMPs levels and miRNA content in Mozambican pregnant women and, secondary, to assess their immunoregulatory role on DCs in vitro. Materials and Methods This study was nested in a placebo-controlled trial of intermittent preventive treatment in pregnancy (IPTp) with sulphadoxine-pyrimethamine (SP) for malaria prevention conducted at the Manhiça Health Research Centre, Mozambique, between 2003–2006 (NCT00209781) . Malaria transmission is perennial with some seasonality and P. falciparum is the predominant species . Women were enrolled at the antenatal clinics of the Manhiça District Hospital if they had gestational age ≤28 weeks, not reported allergies to sulpha drugs and were permanent residents of the study area. After providing written informed consent, women were randomized to receive SP or placebo and screened for HIV if they agreed . The study was approved by the Mozambican National Bioethics Committee and the Hospital Clínic of Barcelona Ethics Review Committee. Maternal HIV-1 infection was diagnosed with Determine HIV-1/2 (Abbott) and confirmed with Unigold rapid test (Trinity Biotech). At delivery, maternal peripheral blood was collected by venipuncture into ethylenediaminetetraacetic acid vacutainers and thin and thick smears prepared, Giemsa-stained and examined for malarial parasites according to quality-control procedures . Blood was centrifuged and plasma stored at -80°C. Placental biopsy specimens were processed for histological examination and classified following published criteria . Newborn's birthweight, gestational age (by Dubowitz method) and maternal hematocrit (in microcapillary tube) were determined. The current analysis was conducted in 61 randomly-selected HIV-infected and 61 non-infected mothers at delivery receiving either placebo or SP from whom demographic and clinical data, as well as maternal plasma samples, were available. Isolation of vesicles from peripheral plasma Plasma samples were thawed at 37°C in water bath and platelets and/or cell debris eliminated by centrifugation (10 min at 1000g, room temperature). Isolation of vesicles from plasma by centrifugation at 10.000g [4,16,34] and ultracentrifugation at 100.000g [21,23] for 30 minutes at 4°C were compared in 8 pregnant mothers from the study and one non-pregnant control (woman from non-endemic area). Centrifugation was chosen as the best approach for the separation. Pelleted vesicles were resuspended in double filtered Dulbecco's Phosphate Buffered Saline without Ca2+ and Mg2+ (DPBS, Lonza). The number of circulating vesicles per μl of plasma was assessed using Perfect-Count Microspheres (Cytognos), a microbead-based single-platform system for absolute counts that includes an internal quality control, according to manufacturer’s protocol. Quantification of total and trophoblast MPs Circulating particles were double stained with Annexin V, that binds to all classes of MPs regardless of cell type , and a mouse monoclonal IgG1 against human Pregnancy-Specific Glycoprotein1 (PSG1, trophoblast marker) for analysis and quantification by flow cytometry. Briefly, 250000 MPs (as assessed with Perfect-Count Microspheres) resuspended in 100 μl of DPBS were incubated with 7 μg/ml PSG1-PE (catalog No. sc-59348, Santa Cruz Biotechnology) for 6 minutes, washed with DPBS, resuspended in 100 μl of Annexin buffer and subsequently stained for 15 minutes with 5 μl Annexin V-FITC (catalog No. 556419 BD Biosciences; and catalog No. ANXVF-200T, Immunostep) for immediate analysis. trMPs were identified in a BD LSRFortessa SORP as events with forward scatter patterns between those obtained with fluorescent calibration microspheres of diameters 0.2 and 1 μm (Invitrogen Molecular Probes) that were positive both for Annexin V and PSG1 staining , and the number of MPs per μl of sample was calculated. Similarly, all the events in this gate positively stained for Annexin V were considered total circulating MPs. Gate limits were set between 0.2 μm to avoid selecting exosomes (up to 0.1 μm ) and 1 μm to avoid selecting apoptotic bodies (>1 μm ). The gating strategy is shown in S1 Fig. Negative staining with uranyl acetate was used for electron microscopy imaging. Briefly, 5 μl of isolated MP sample were loaded in a previously charged metal grid, deposited for 30 min, fixed with 1% glutaraldehyde for 5 minutes, washed with water, stained with 2% uranyl acetate for 1 minute and imaged with JEOL 1010 TEM (JEOL). Analysis of placental microRNA content by qPCR Plasmas from 10 mothers negative both for malaria and HIV, 10 placental malaria-infected and 10 HIV-infected women with similar characteristics to the study population (described in S1 Table) and that presented the highest concentrations of trMPs were selected. No double infections were included. Additionally, plasmas from 5 men from non-endemic area, not infected by malaria neither HIV, were included as negative controls. Potential hemolysis in plasma samples (1:10 diluted), shown to greatly affect miRNA expression , was addressed by spectrophotometric analysis using the Harboe’s method with Allen’s correction to calculate hemoglobin concentration. Absence of hemolysis was defined by an absorbance of less than 0.2 at 414 nm . Four human placental miRNAs were selected because of their specific profile in the placenta or maternal plasma, their importance in regulatory processes during the gestation and their participation in several pregnancy disorders: hsa-miR-21, highly expressed at the end of pregnancy; hsa-miR-493, representative of chromosome 14 miRNA cluster (C14MC) whose expression decreases at the end of pregnancy; hsa-miR-517c and hsa-miR-519d, both from the cluster C19MC and highly expressed at the end of pregnancy [26,40,41]. In addition, hsa-miR-191 was included as a plasma normalization control . Circulating MPs were isolated from 200 μl of plasma as previously described. RNA was extracted with miRCURY™ RNA Isolation Kit–Biofluids (Exiqon) following manufacturer’s protocol. Reverse transcription was performed with miRNA specific stem-loop RT primers and TaqMan MicroRNA Reverse Transcription Kit (Applied Biosystems) using 10 ng of total RNA measured by Nanodrop (Thermo Scientific) and following the protocol provided by the supplier. qPCR with specific TaqMan Assays (hsa-miR-21, assay ID 000397; hsa-miR-493-5p, assay ID 001040; hsa-miR-517c, assay ID 001153; hsa-miR-519-3p, assay ID 002403; hsa-miR-191-5p, assay ID 002299) and TaqMan Universal PCR Master Mix (all from Applied Biosystems) was performed using 1.5 μl of reverse transcription product per reaction. All reactions were run in duplicates including no-template controls in 96-well plates on a Mx3005P QPCR System (Applied Biosystems) and threshold cycles (Cts) were obtained after 40 cycles. If miRNA expression was not detected in a sample, a Ct value of 41 was arbitrary assigned. ΔΔCts were calculated using values from non-infected women as calibrators and miR-191 as endogenous control . Total P. falciparum RNA was included as a negative control. In vitro stimulation of dendritic cells with MPs Maternal antigen-presenting cells, including DCs, are important players in this fetal-maternal immune adjustment between stimulatory and tolerogenic responses and are involved in cell-cell interactions in the decidualized endometrium . In addition, DCs have been used in previous MP studies as a model to assess the effect of EVs on immune cell [44,45]. Human DCs were generated from monocytes obtained from buffy coats of healthy donors that provided written informed consent, and grown in serum added-X-VIVO-15 culture medium (Sigma and Lonza, respectively) with interleukin 4 (IL4) and granulocyte macrophage colony-stimulating factor (GM-CSF) (both from Miltenyi Biotec). To evaluate the immunoregulatory properties of MPs, 100.000 DCs in 100 μl of culture medium were incubated for 24h with total MPs obtained from 20 μl plasma of individual women: 8 negative both for malaria and HIV (MPNI), 10 HIV-infected (MPHIV) and 8 women with placental malaria by histology (MPmalaria) with similar characteristics to the study population and that presented the highest concentrations of trMPs in plasma. No double infections were included. DCs were incubated in every experiment with 400 ng of lipopolysaccharide (LPS) (Invivogen) as a positive and normalization control, and with cell culture medium alone as a negative control. To determine whether MPs have a synergistic effect on LPS mediated DC-stimulation, a second incubation condition with the same amount of MPs plus 10 ng of LPS was included for each sample. DC phenotype after in vitro stimulation with MPs was analysed using the co-stimulatory and activation markers CD80, CD86 and major histocompatibility complex II (MHCII). Cells were incubated with 1μg/ml of anti-CD80, anti-CD86 and anti-MHCII (BD Biosciences) during 30 minutes at 4°C, washed and stained with secondary polyclonal goat anti-mouse-PE antibody (catalog No. 550589, BD Biosciences) for 30 minutes at 4°C. Fluorescence measurements were performed by flow cytometry in a BD FACSCantoII. Normalization of percentages and mean fluorescence intensities (MFIs) of activation markers with respect to the positive control was performed by arbitrary assigning the value 100% to percentages and 1 to MFIs for the LPS condition in each experiment in order to control for potential technical and donor variabilities. Cytokine and chemokine concentrations were analysed in 50 μl of post-stimulus culture supernatant using the Cytokine Human 30-plex Panel (Life Technologies) as indicated by the manufacturer. Plates were read with a Luminex xMAPTM (Luminex) and concentrations calculated with respect to the kit standards. Concentrations over or below the limit of quantification of the standard curves were assigned twice the value of the most concentrated standard or half the value of the most diluted one, respectively. The available volume of culture supernatant did not allow including several dilutions, and therefore undiluted supernatants were used for the assay. Pregnant women were classified as primigravidae (first pregnancy), secundigravidae (second pregnancy) and multigravidae (≥2 previous pregnancies). Age was categorized as ≤20, 20–24 or ≥25 years on the basis of maternal age terciles in this population. Placental and peripheral malaria infections were defined by the presence of parasites by histology in placental sections (active placental infection) or by microscopy in peripheral blood, respectively. Maternal anemia was considered if the hematocrit level was <33%. Placental inflammation was defined as >5 mononuclear inflammatory cells observed by histological examination in 10 high-power fields using 400x magnification . Low birthweight (LBW) was considered for newborns of less than 2500g, and preterm delivery for gestational ages <37 weeks. trMPs concentration was compared between isolation methodologies using the Wilcoxon signed rank test. Student’s t test and linear regression models were used to estimate the association of HIV, malaria and other clinical and demographic covariates with log-transformed MP levels, percentages and MFIs of DC activation markers, and cytokine/chemokine concentrations in culture supernatants. Logistic regression models were used to estimate associations between adverse pregnancy outcomes (placental inflammation, preterm delivery, LBW and maternal anemia) and MP levels. Multivariate models were adjusted for maternal HIV, peripheral and placental malaria infection, parity, age and IPTp group. miRNA levels expressed as ΔΔCt were compared between infection groups by Student's t test. Statistical analysis was performed using GraphPad Prism version 6 (GraphPad Software) and Stata version 11.0 (StataCorp). Characteristics of the study population The prevalence of peripheral P. falciparum infection was 15.6% (19/122). Parasites were found in 23 placental sections by histology (18.8% active infections). In total, 26 (21.3%) women presented parasites in one or both compartments by any of the techniques at the time of delivery. The characteristics of the 122 women at delivery are presented in Table 1. Fifty-six (45.9%) women received placebo and 66 (54.1%) women received SP. The subset of 122 included in this study and the 1030 women participating in the randomized trial were comparable in terms of parity, age, ITPp group, peripheral and placental malaria infection. Women with active placental malaria were younger, had lower parity and presented a higher risk of suffering peripheral malaria than non-infected. No differences were found in anemia, HIV and IPTp intervention between malaria-infected and non-infected mothers (Table 1). There were no significant differences between HIV infected and non-infected women by IPTp treatment received (P = 0.467). Characterization of total and trophoblast MPs No significant difference was found in the number of trMPs per μl of plasma between centrifuged or ultracentrifuged samples (P = 0.500). However, more clearly separated Annexin V-positive populations were obtained by 10.000g centrifugation (S2 Fig), in line with previous methodologies for MP separation . A high-positive Annexin V population was observed in the centrifugated samples but not when ultracentrifugation was applied, which might be explained by MP lysis when high g-force is applied (S2 Fig). By electron microscopy, it was confirmed that Annexin V-positive MPs are a population of membranous vesicles of heterogeneous size (0.2–1 μm) and morphology (Fig 1). Association of circulating MP levels with infection and pregnancy outcomes Numbers of total (Fig 2A) and trMPs (Fig 2B) per μl of plasma for the 122 women were compared according to their infectious status. In a univariate analysis, concentrations of total MPs and trMPs were higher in HIV-infected than in non-infected women (P = 0.008 and P = 0.040, respectively), and the significant differences were maintained in the multivariate analysis (1.99-fold increase, 95% CI 1.24–3.21, P = 0.005; and 1.77-fold increase, 95% CI 1.06–2.96, P = 0.030, respectively). Concentrations measured by flow cytometry are expressed as number of MPs per μl of plasma. Horizontal lines represent the geometric mean and the 95% confidence interval. P-values were obtained from the univariate analysis. Significant differences maintained in the multivariate analysis are indicated by an asterisk. No significant differences were found between women with and without placental malaria in the number of total MPs (P = 0.656) or trMPs (P = 0.910) per μl of plasma. Similarly, no significant differences were found in total MP (P = 0.851) or trMP (P = 0.669) concentration between women with or without peripheral malaria, neither among age (total MPs: P = 0.434 and P = 0.626 for age groups 20–24 or ≥25 compared with <20, respectively; and trMPs: P = 0.341 and P = 0.567 for age groups 20–24 or ≥25 compared with ≤20, respectively) or parity groups (total MPs: P = 0.692 and P = 0.677 for secundigravidae or multigravidae compared with primigravidae, respectively; and trMPs: P = 0.568 and P = 0.892 for secundigravidae or multigravidae compared with primigravidae, respectively). The association between pregnancy outcomes and MP concentrations was analysed in a univariate model. Women delivering LBW babies had higher levels of total MP (P = 0.032) and trMP (P = 0.021) than mothers delivering normal birthweight newborns (Fig 3). The association was maintained in the multivariate analysis both for concentrations of total MP (odds ratio, OR = 2.38 with doubling levels of MPs; 95% CI, 1.08–5.25; P = 0.032) and trMP (OR = 2.17; 95% CI, 1.06–4.46; P = 0.034), and was therefore independent from maternal HIV or malaria infections. No associations with maternal anemia (53 women), placental inflammation (66 women) or preterm delivery (3 women) were found. Concentrations are expressed as number of MPs per μl of plasma. Horizontal lines represent the geometric mean and the 95% confidence interval. P-values were obtained from the univariate analysis. Significant differences maintained in the multivariate analysis are indicated by an asterisk. Placental miRNA content of circulating MPs The 30 women included in the miRNA sub-study were comparable with the whole study cohort in terms of age, parity, malaria and HIV infections, IPTp, anemia and low birthweight babies, with the exception of placental inflammation that was more prevalent in the whole cohort (S1 Table). No significant hemolysis was observed in any of the 30 plasma samples used for miRNA quantification [Median A414 = 0.081, interquartile range, IQR 0.067–0.105; median haemoglobin concentration (g/L) = 0.153, IQR 0.052–0.318]. miR-191 and miR-21 were detected in all MP samples isolated from pregnant women. miR-517c was detected in 87.5% (7/8) of MPs from women with active placental malaria, compared with 41.7% (5/12) of non-infected (P = 0.040) and 50% (5/10) of HIV-infected women (P = 0.093). miR-519d was expressed in MPs from 66.7% of non-infected, 75% of active malaria and 70% of HIV infections (P = 0.924). miR-493 expression was only detected in 8% of non-infected, 25% of mothers with placental malaria and 10% of women with HIV (P = 0.522) and not further included in the analysis. Raw Cts by infection status are shown in S3 Fig. When ΔΔCts for each miRNA normalized with respect to miR-191 were compared among infected and non-infected women (Fig 4), miR-517c expression was found to be higher in MPs from mothers with active placental malaria than in the non-infected group (P = 0.035). MicroRNA expression levels were calculated with the delta delta Ct method. Non-infected women microparticle results were used as calibrators and miR-191 as endogenous control. Twelve women were included in the negative group (negative both for malaria and HIV), 8 in the active placental malaria and 10 in the HIV-infected group. Five men were included as negative controls for the expression of placental-related microRNAs. Means and standard deviations are represented. Statistically significant differences from Student's t test are marked with an asterisk. miR-493 was only expressed in 4/30 (13.3%) of the women samples, and therefore not included in the analysis by maternal infection status. There were no significant differences in miRNA expression between women with or without placental inflammation (P = 0.581 for miR-21, P = 0.695 for miR-517c and P = 0.989 for miR-519d). Only one woman delivering a LBW baby was included in the miRNA study, and therefore a comparison between LBW and normal birthweight delivering mothers was not possible. No statistically significant correlations were found between birthweight as a continuous variable and miRNA expression. Functional effect of trophoblast MPs on dendritic cells After incubation for 24h with MPs isolated from 26 plasmas, DCs expressed the activation markers CD80, CD86 and MHCII (Fig 5), although there were no significant differences in the expression of CD80 and CD86 between DCs incubated with MPNI, MPmalaria or MPHIV. In the univariate analysis, a higher percentage of DCs incubated with MPHIV expressed MHCII (P = 0.049) and levels of MHCII expression were higher (P = 0.021), when compared with DCs stimulated with MPNI. After adjustment, the percentage of DCs expressing MHCII remained significantly augmented when incubating with MPHIV (1.05-fold increase; 95% CI, 1.00–1.10; P = 0.041) compared to MPNI, and the MHCII MFI was increased but with borderline significance (1.22-fold increase; 95% CI, 0.988–1.51; P = 0.063). The normalized percentage of dendritic cells expressing an activation marker (a) or the normalized mean fluorescence intensity (MFI) (b) are represented as circles (negative both for malaria and HIV), squares (active malaria-infected) or triangles (HIV-infected). Percentages and MFIs were normalized with respect to the positive control incubated with lipopolysaccharide. Geometric means and 95% confidence intervals are represented. P-values correspond to the univariate analysis. Statistically significant differences maintained after adjustment in the multivariate model are marked with an asterisk. Several cytokines and chemokines were detected in DC culture supernatants after incubation with MPs, although, in general, there were not significant differences among MPNI, MPmalaria or MPHIV alone (Fig 6) or after simultaneous incubation of MPs with LPS (S4 Fig). Only MPHIV induced lower production of Monocyte Chemoattractant Protein-1 (MCP1) in DC supernatants (0.539-fold decrease; 95% CI, 0.346–0.838; P = 0.008) than MPNI in the univariate analysis, and the difference was maintained after adjustment (0.426-fold decrease; 95% CI, 0.219–0.827; P = 0.015). There was a trend towards a higher expression of IL13 when incubating DCs with MPs and LPS than with LPS alone, although not statistically significant. Twenty-six women were included. Concentrations are expressed in pg/ml. PC represents the positive control condition with only lipopolysaccharide. Geometric means and 95% confidence intervals are represented. P-values correspond to the univariate analysis. Statistically significant differences maintained after adjustment in the multivariate model are marked with an asterisk. This is the first study that, to our knowledge, analyses the relationship of malaria and HIV infection in pregnancy with alterations in circulating MPs and their miRNA content. Total and trMPs were increased in women infected with HIV, which in vitro led to increased expression of MHCII and reduced MCP1 in DCs, and in mothers delivering LBW babies, compared to MPs from non-infected or those delivering normal birthweight newborns, respectively. Although malaria in pregnant women was not associated with changes in circulating MPs, the expression of pregnancy-related microRNAs (miR-517c) in MPs was higher in active placental malaria than in uninfected mothers. Overall, it is plausible to speculate a role for trMPs and placental miRNAs in the pathophysiology of malaria and HIV infections during pregnancy. The augmented levels of total MPs in HIV-positive women could be explained by platelet activation and increase of procoagulant MPs associated with HIV infection [47,48]. HIV-related changes in placental membranes might lead to increased concentration of trMPs and contribute to the augmented risk of LBW reported among HIV-infected pregnant women . In line with previous reports showing increased levels of MPs, including placental MPs, in pre-eclampsia [23,50], IUGR and recurrent miscarriage , as well as in endothelial activation and induction of a procoagulant state , the present results support a crucial role for MPs in placenta dysfunction and vascular pregnancy complications . Similar mechanisms may contribute to MP-dependent LBW in HIV-infected pregnant women. As placental MPs gradually increase from approximately 10 weeks of gestation , their measurement might be useful for the identification of pregnancies with LBW risk at early stages of gestation. The increase of placental MPs associated with inflammation in pathologies such as pre-eclampsia was not observed in women with placental inflammation from our cohort. The effect of placental inflammation on MP release should be addressed in future studies analysing cytokines and other inflammatory mediators in the mothers. In contrast to HIV, placental malaria was not associated with changes in MP concentrations, suggesting placental alterations and molecular mechanisms different than those observed for MPs in HIV infection. However, malaria may be associated with changes in vesicle miRNA content. miR-517c from the cluster C19MC, previously shown to have a role as immunomodulator in pregnancy and tumorigenesis , was found overexpressed in mothers with placental malaria compared with non-infected. Our study suggests an interesting diagnostic potential for pregnancy-specific miRNAs from C19MC , in line with reports showing an augment of miR-517c expression with increasing placental weight and an overexpression in pre-eclampsia and recurrent spontaneous abortion , that should be addressed in future studies. MPs from HIV-infected mothers induced a higher expression of MHCII in DCs than MPs from uninfected women, suggesting a modulatory effect in DC function. Moreover, the increased MHCII expression in DCs after incubation with MPs from HIV-infected women without upregulation of co-stimulatory molecules, as well as the trend towards a higher expression of IL13 in DCs after incubation with MPs compared with LPS alone, is in line with the previously proposed tolerogenic role of MPs in pregnancy [21,22]. Although the changes induced by MPs in our DC system are modest and their biological relevance needs confirmation, they are indicative of an in vitro effect over immune cells even when only 20 μl of plasma are used as starting material. Higher numbers of MPs are expected to have increased stimulatory capacity and, therefore, a more pronounced effect over DCs that should be determined in future studies. Our study presents several limitations. First, EVs are a highly heterogeneous population, making difficult to completely discard some contaminations with exosomes, apoptotic bodies or other plasma components during MP preparation. Since the miRNA analysis was performed in a small number of women and miRNAs, further studies are needed including more samples and profiling methodologies. Other potential maternal infections, endogenous factors or environmental exposures that could affect MP release or miRNA expression should be taken into account in future works. Since we chose to incubate DCs with MPs obtained from the same volume of plasma from all the participants to reproduce physiological conditions where circulating MP concentrations are variable, the effect of normalized amounts of MPs was not addressed. Moreover, we did not correct for multiple comparisons when assessing the effect of MPs on cytokine/chemokine production by DCs, although in general there were no significant differences after incubation with MPNI, MPmalaria or MPHIV, with the exception of the decrease in MCP1 with MPHIV, that was consistently maintained in the multivariate analysis. Finally, the cross-sectional nature of this work describes potential associations but causal relationships cannot be inferred. In summary, this study shows that plasma levels of total and trophoblast-derived MPs, which are increased in HIV-infected pregnant women and in those delivering LBW newborns, can alter the in vitro function of immune regulatory cells such as DCs. In contrast, active placental malaria is associated with changes in miRNA of placental origin contained in circulating MPs. Both MPs and miRNAs might affect the immunoregulatory balance during pregnancy, although further studies to unravel the mechanisms are required. Importantly, the results of this study suggest that placental miRNAs might present a utility as biomarkers of placental malaria infection during pregnancy that should be further explored and validated. The characterization of different vesicle populations and their content may increase our understanding of immunopathological processes in malaria and HIV and guide the discovery of new biomarkers for malaria in pregnancy, an infection that frequently remains undetected but still has deleterious consequences . S1 Fig. Gating strategy for the quantification of total and trophoblast microparticles. A. Regions corresponding to calibration beads of 0.2, 0.5 and 1 μm of diameter are shown. B. All the events in the sample indicating the regions for the calibration beads and the background (BG). C. Events with forward scatter patterns (FSC-A) between those obtained with fluorescent calibration microspheres of diameters 0.2 and 1 μm (left) positively stained for Annexin V (right) were considered total circulating microparticles (in this particular example, 89.6% of the total events after background exclusion). D. Unstained control sample showing the gate for the annexin V and PSG1 positive population. E. Events that were positive both for Annexin V and PSG1 staining were considered trophoblast microparticles (30.4% of the population of particles 0.2–1 μm in the example). S2 Fig. Comparison of circulating microparticles isolation methodologies. Example of gating and percentages of the Annexin V-positive population after isolation of circulating microparticles from plasma by centrifugation (CENT) at 10.000g (A, unstained control; B, annexin V stained sample) or ultracentrifugation (ULTRA) at 100.000g (C, unstained control; D, annexin V stained sample). S3 Fig. Raw cycle thresholds (Cts) of placental microRNAs determined by qPCR in circulating microparticles samples. Non-infected women are represented by circles, women with active placental malaria by squares and HIV-positive mothers by triangles. Means and standard deviations are represented. A value of Ct = 41 was arbitrarily assigned to samples where the expression of a microRNA was not detected after 40 qPCR cycles. S4 Fig. Cytokine (a) and chemokine (b) concentrations in dendritic cell culture supernatants after incubation with microparticles plus lipopolysaccharide. Twenty-six women were included. Concentrations are expressed in pg/ml. PC represents the positive control condition with only lipopolysaccharide. Geometric means and 95% confidence intervals are represented. Statistical analysis did not reveal statistical differences among the groups and therefore P-values are not indicated (P≥0.005). S1 Table. Comparison of demographic and clinical factors between mothers included in the microRNA subset and the whole study cohort. The authors are grateful to the study participants and to the staff at the Manhiça District Hospital and Centro de Investigaçao em Saúde de Manhiça for their continued dedication and collaboration. The authors are particularly grateful to Gemma Moncunill, Pilar Requena and Ruth Aguilar for their helpful advice; and to Isabel Crespo and Cristina López from the IDIBAPS Flow Cytometry and Cell Sorting Facility for their participation in vesicle analysis. Conceived and designed the experiments: LM AB EM DMP IM BS CD URM DBR PLA CM AM. Performed the experiments: LM DB DMP CE. Analyzed the data: LM AB EM DB DMP CE IM BS CD URM DBR PLA CM AM. Contributed reagents/materials/analysis tools: AB EM IM BS CD URM DBR PLA CM AM. Wrote the paper: LM AB EM DB DMP CE IM BS CD URM DBR PLA CM AM. - 1. Delabranche X, Berger A, Boisramé-Helms J, Meziani F. 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Schwartz DA, Sungkarat S, Shaffer N, Laosakkitiboran J, Supapol W, Charoenpanich P, et al. Placental abnormalities associated with human immunodeficiency virus type 1 infection and perinatal transmission in Bangkok, Thailand. J Infect Dis. 2000 Dec;182(6):1652–7. pmid:11069236 - 32. Menéndez C, Bardají A, Sigauque B, Romagosa C, Sanz S, Serra-Casas E, et al. A randomized placebo-controlled trial of intermittent preventive treatment in pregnant women in the context of insecticide treated nets delivered through the antenatal clinic. PLOS One. 2008 Jan;3(4):e1934. doi: 10.1371/journal.pone.0001934. pmid:18398460 - 33. Alonso PL, Smith T, Schellenberg JR, Masanja H, Mwankusye S, Urassa H, et al. Randomised trial of efficacy of SPf66 vaccine against Plasmodium falciparum malaria in children in southern Tanzania. Lancet. 1994 Oct;344(8931):1175–81. pmid:7934537 - 34. Guller S, Tang Z, Ma YY, Di Santo S, Sager R, Schneider H. Protein composition of microparticles shed from human placenta during placental perfusion: Potential role in angiogenesis and fibrinolysis in preeclampsia. Placenta. 2011 Jan;32(1):63–9. doi: 10.1016/j.placenta.2010.10.011. pmid:21074265 - 35. Camolotto S, Racca A, Rena V, Nores R, Patrito LC, Genti-Raimondi S, et al. Expression and transcriptional regulation of individual pregnancy-specific glycoprotein genes in differentiating trophoblast cells. Placenta. 2010 Apr;31(4):312–9. doi: 10.1016/j.placenta.2010.01.004. pmid:20116096 - 36. Redman CW, Tannetta DS, Dragovic RA, Gardiner C, Southcombe JH, Collett GP, et al. Review: Does size matter? Placental debris and the pathophysiology of pre-eclampsia. Placenta. 2012 Feb;33 Suppl:S48–54. doi: 10.1016/j.placenta.2011.12.006. pmid:22217911 - 37. Hristov M, Erl W, Linder S, Weber PC. Apoptotic bodies from endothelial cells enhance the number and initiate the differentiation of human endothelial progenitor cells in vitro. Blood. 2004 Nov 1;104(9):2761–6. pmid:15242875 - 38. Kirschner MB, Kao SC, Edelman JJ, Armstrong NJ, Vallely MP, van Zandwijk N, et al. Haemolysis during sample preparation alters microRNA content of plasma. PLOS One. 2011 Jan;6(9):e24145. doi: 10.1371/journal.pone.0024145. pmid:21909417 - 39. Harboe M. A method for determination of hemoglobin in plasma by near-ultraviolet spectrophotometry. Scand J Clin Lab Invest. 1959 Jan;11:66–70. pmid:13646603 - 40. Miura K, Miura S, Yamasaki K. Identification of Pregnancy-Associated MicroRNAs in Maternal Plasma. Clin Chem. 2010;56(11):1767–71. doi: 10.1373/clinchem.2010.147660. pmid:20729298 - 41. Kotlabova K, Doucha J, Hromadnikova I. Placental-specific microRNA in maternal circulation- identification of appropriate pregnancy-associated microRNAs with diagnostic potential. J Reprod Immunol. 2011 May;89(2):185–91. doi: 10.1016/j.jri.2011.02.006. pmid:21513988 - 42. Peltier HJ, Latham GJ. Normalization of microRNA expression levels in quantitative RT-PCR assays: identification of suitable reference RNA targets in normal and cancerous human solid tissues. RNA. 2008 May;14(5):844–52. doi: 10.1261/rna.939908. pmid:18375788 - 43. Laskarin G, Kämmerer U, Rukavina D, Thomson AW, Fernandez N, Blois SM. Antigen-presenting cells and materno-fetal tolerance: an emerging role for dendritic cells. Am J Reprod Immunol. 2007 Sep;58(3):255–67. pmid:17681042 - 44. Bastos-Amador P, Pérez-Cabezas B, Izquierdo-Useros N, Puertas MC, Martinez-Picado J, Pujol-Borrell R, et al. Capture of cell-derived microvesicles (exosomes and apoptotic bodies) by human plasmacytoid dendritic cells. J Leukoc Biol. 2012 May;91(5):751–8. doi: 10.1189/jlb.0111054. pmid:22319103 - 45. Nomura S, Fujita S, Nakanishi T, Yokoi T, Shimamoto K, Miyamoto R, et al. Platelet-derived microparticles cause CD154-dependent activation of dendritic cells. Platelets. 2012 Jan;23(1):81–2. doi: 10.3109/09537104.2011.595850. pmid:22150414 - 46. Naniche D, Serra-Casas E, Bardají A, Quintó L, Dobaño C, Sigauque B, et al. Reduction of antimalarial antibodies by HIV infection is associated with increased risk of Plasmodium falciparum cord blood infection. J Infect Dis. 2012 Feb 15;205(4):568–77. doi: 10.1093/infdis/jir815. pmid:22238468 - 47. Mayne E, Funderburg NT, Sieg SF, Asaad R, Kalinowska M, Rodríguez B, et al. Increased platelet and microparticle activation in HIV infection: upregulation of P-selectin and tissue factor expression. J Acquir Immune Defic Syndr. 2012 Apr 1;59(4):340–6. doi: 10.1097/QAI.0b013e3182439355. pmid:22156911 - 48. Baker J V, Huppler Hullsiek K, Bradford RL, Prosser R, Tracy RP, Key NS. Circulating levels of tissue factor microparticle procoagulant activity are reduced with antiretroviral therapy and are associated with persistent inflammation and coagulation activation among HIV-positive patients. J Acquir Immune Defic Syndr. 2013 Jul 1;63(3):367–71. doi: 10.1097/QAI.0b013e3182910121. pmid:23507662 - 49. Markson LE, Turner BJ, Houchens R, Silverman NS, Cosler L, Takyi BK. Association of maternal HIV infection with low birth weight. J Acquir Immune Defic Syndr Hum Retrovirol. 1996 Nov 1;13(3):227–34. pmid:8898667 - 50. Lok CA, Van Der Post JA, Sargent IL, Hau CM, Sturk A, Boer K, et al. Changes in microparticle numbers and cellular origin during pregnancy and preeclampsia. Hypertens Pregnancy. 2008 Jan;27(4):344–60. doi: 10.1080/10641950801955733. pmid:19003636 - 51. Aharon A, Brenner B. Microparticles and pregnancy complications. Thromb Res. 2011 Feb;127 Suppl: S67–71. - 52. Alijotas-Reig J, Palacio-Garcia C, Llurba E, Vilardell-Tarres M. Cell-derived microparticles and vascular pregnancy complications: a systematic and comprehensive review. Fertil Steril. 2013 Feb;99(2):441–9. doi: 10.1016/j.fertnstert.2012.10.009. pmid:23122952 - 53. Toth B, Jeschke U, Rogenhofer N, Scholz C, Würfel W, Thaler CJ, et al. Recurrent miscarriage: current concepts in diagnosis and treatment. J Reprod Immunol. 2010 May;85(1):25–32. doi: 10.1016/j.jri.2009.12.006. pmid:20185181 - 54. Bullerdiek J, Flor I. Exosome-delivered microRNAs of “chromosome 19 microRNA cluster” as immunomodulators in pregnancy and tumorigenesis. Mol Cytogenet. 2012 Jan;5(1):27. doi: 10.1186/1755-8166-5-27. pmid:22559272 - 55. Miura K, Morisaki S, Abe S, Higashijima A, Hasegawa Y, Miura S, et al. Circulating levels of maternal plasma cell-free pregnancy-associated placenta-specific microRNAs are associated with placental weight. Placenta. 2014 Jun 26;35(10):848–51. doi: 10.1016/j.placenta.2014.06.002. pmid:25015175 - 56. Dong F, Zhang Y, Xia F, Yang Y, Xiong S, Jin L, et al. Genome-wide miRNA profiling of villus and decidua of recurrent spontaneous abortion patients. Reproduction. 2014 Jul;148(1):33–41. doi: 10.1530/REP-14-0095. pmid:24686457 - 57. 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<urn:uuid:ff861ba8-49ad-489e-a6fd-6c35e9480205>	I have rugs and mats on the floor. I guess they help a little. But my kitchen floor still gets dirty. I tried swiffering every day, even twice a day. It stays clean for a few hours. The true test of a clean floor: Try Wearing White Socks! I want my white socks to stay clean on the bottoms. I walk across my kitchen floor, and my white socks are already dirty. I take them off and drop them in the laundry basket. Worn for about five minutes, and they're laundry fodder. I don't like to wear shoes in the house. I like socks. I like white socks. White socks that stay clean on the bottoms make me feel good and happy. Clean white socks in the house. My dream. Isn't there a Dr. Seuss verse for white socks in the house?
<urn:uuid:e7a7ce65-6a2e-4f47-9ef2-50f7aced08a1>	The Jowett Club of Australia provides a free service for members to advertise vehicles for sale. proceeding please note the bulleted points club makes every endeavour to ensure the advertisements are accurate, however the club accepts no responsibility for the accuracy of the vehicle's description, or that the seller actually owns the vehicle. The club recommends that each potential buyer makes a comprehensive inspection of the vehicle and checks the ownership of the vehicle offered for sale. The club takes no part in the sale of any vehicle, it only provides a "bill board" on which advertisements are placed. down to looking at the vehicles for sale. Scroll down to see all vehicles. At the very bottom of this page there is a form which will let you upload a potential advertisement for this page. The Club will review the advertisement before
<urn:uuid:6b998113-2cea-475a-92c7-44d75c452857>	I am originally from South Africa, have had a passion for photography all my life, trying to bring everything I enjoy into my art. I enjoy photographing anything out of the ordinary, things normal people dont know about or ever get the chance to see, sometimes ecompassing an element of danger. I currently live in LA and it has to be one of the most diverse places I have ever been to, it also allows me to shoot everything from Sport, Documentary's and Portraits and occasionally Press work. Find me at: http://www.photobrett.com 13 Sep 2012 — 95 views, 1 favorite In "Photo Challenge: Sports in Motion" 13 Sep 2012 — 76 views, 2 favorites 7 Jun 2010 — 180 views, 2 favorites 4 Jun 2010 — 79 views, 1 favorite 3 Jun 2010 — 338 views Collections are a JPG+ feature. You must be a JPG+ member to create new collections and to add photos to collections. Sign up for JPG+ to start using collections now! 344 Views » 0 Comments » 0 Votes 387 Views » 0 Comments » 0 Votes 634 Views » 0 Comments » 0 Votes Q: What is the best lens for theater photography? A: Do you know?
<urn:uuid:a10312f3-ba3b-4acf-8ddc-f86532d7ee08>	IB in WT Update INDIANAPOLIS, IN (January 9, 2017) – Having been authorized as International Baccalaureate (IB) World Schools in 2013, all seven Washington Township elementary schools have recently undergone their first official International Baccalaureate Primary Years Program (PYP) evaluation. Washington Township is pleased to report that none of the schools received matters to address indicating that ALL elementary schools are upholding the integrity of the IB program. Being evaluated by the International Baccalaureate Organization allows Washington Township to realize how well the district is meeting the standards and practices that IB has established as part of their quality assurance framework. There are seven standards and approximately one hundred practices regarding philosophy, organization, and curriculum that are implemented by IB schools all over the world. The framework for the IB evaluation is built in such a way as to recognize each school’s strengths (commendations) and at the same time, explore areas for improvement (recommendations). “The Washington Township elementary schools consistently received a good ratio of commendations to recommendations,” said Mr. Chad Hyatt, Washington Township Curriculum Coach. “One of the common commendations across the board was the commitment of the governing body of the school district to the implementation of the IB program. One of the common recommendations across all seven elementary schools was a recommendation that is seen frequently among ALL IB schools, and that is to improve the transdisciplinary aspect of the program. The PYP is committed to students learning about their world through the integration of subject areas, just like they naturally do in life. With mandates from the state for segmented instruction in subjects like literacy and math this can often be a challenge especially for public schools to overcome. As a district, we are constantly exploring ways to keep this philosophy at the forefront of our efforts so that we meet our state standards, satisfy state requirements, and implement an IB program within a transdisciplinary focus,” concluded Hyatt. The International Baccalaureate Organization does not run or operate schools, but instead they work with schools to offer quality international education. Washington Township elementary schools will not be evaluated again for five years. The three middle schools and North Central High School will begin their first evaluation process in the 2018-19 school year with an official visit from IB the following year. For more information about the International Baccalaureate programs in Washington Township, visit http://www.msdwt.k12.in.us/academic-programs/
<urn:uuid:7e915dfe-82a7-4aa5-9325-32134c5c42b7>	The Olympic Torch Relay will start its 10-mile tour through three parts of Juneau at 8:30 a.m. Thursday at the Juneau Airport. It will be the first time the Olympic flame has burned in Alaska. There will be several key vantage points for residents to view the torch as it makes its way through the Mendenhall Valley, Lemon Creek area and downtown Juneau. Times listed are estimates of when the torch will pass key spots and could vary by as much as 10 minutes. A caravan passing out pennants and other goodies from torch relay sponsor Coca-Cola will lead the main procession by about 20 minutes. Maps of the Juneau course ran in the Juneau Empire's Olympic Torch Relay special section Jan. 16 and are posted on the torch relay link on the Juneau Empire's Web site, www.juneauempire.com. Residents might have to wait through a few rolling traffic stops as the torch makes its way through town, but the only places where there will be full stops are in front of the airport at the start of the relay, on the uphill side of the Ninth Street hill downtown and near the Governor's Mansion. Recommended parking areas are at Dimond Park, Brotherhood Bridge, Western Auto, Twin Lakes and the downtown subport. The first part of the course will run through the Mendenhall Valley. The torch is expected to pass Glacier Valley Elementary School about 9 a.m., Riverbend School about 9:10 a.m. and end the segment at the Brotherhood Bridge parking lot about 9:25 a.m. The second part of the course starts at sponsor Capital Chevrolet in Lemon Creek about 9:30 a.m. The torch will pass by the Juneau Pioneers' Home about 9:40 a.m. and end at the Salmon Creek Medical Center near Twin Lakes at 9:55 a.m. The third segment starts at the Glacier Highway entrance off Egan Drive about 10 a.m. and will head toward downtown Juneau. The torch will pass by the Breakwater Inn and Juneau-Douglas High School about 10:10 a.m., the Governor's Mansion about 10:20 a.m. and will reach the Goldbelt Dock about 10:30 a.m. From the Goldbelt Dock, the torch will be carried across Juneau Harbor by Ethel Lund in a Tlingit canoe paddled by nine members of the Tlingit Warriors canoe team and two members of the SouthEast Alaska Regional Health Consortium canoe team. The Tlingit dance group Yun Shu Ka will sing an exit song as the canoe takes off, then will be bussed to the Intermediate Vessel Float Dock near Taku Smokeries to sing an entrance song as the canoe comes back to shore about 10:40-10:45 a.m. The public will not be allowed on the Goldbelt Dock for safety reasons and the best views of the canoe segment are along the city dock and behind the Hangar on the Wharf. The torch will then be carried through downtown, passing the Capitol about 11 a.m. and entering Centennial Hall for the citywide celebration about 11:10 a.m. Doors will open at about 10 a.m. and the celebration will last until noon. Viewers are encouraged to carpool to reduce parking problems, especially since snow is predicted. The city will tow illegally parked cars and drivers should not park on relay route streets or roadsides. © 2017. All Rights Reserved. \| Contact Us
<urn:uuid:0fcd9629-ccf6-4c61-8366-fd690c9ecd8b>	Today, April 16 Juneau-Douglas High School boys and girls soccer - Spring Fever event: The Crimson Bear boys and girls host teams from Chugiak, Palmer and Pagosa Springs (Colo.) at Adair-Kennedy Memorial Park. Chugiak boys play Palmer, 2:30 p.m.; Chugiak girls vs. Pagosa Springs, 4:15 p.m.; Juneau girls vs. Palmer, 6:15 p.m.; Juneau boys vs. Palmer, 8 p.m. Saturday, April 17 Juneau-Douglas High School boys and girls soccer - Spring Fever event: The Crimson Bear boys and girls host teams from Chugiak, Palmer and Pagosa Springs (Colo.) at Adair-Kennedy Memorial Park. Juneau JV girls vs. All-Stars, 10 a.m.; Juneau boys vs. Chugiak, 1 p.m.; Palmer girls vs. Pagosa Springs, 2:45 p.m.; Juneau boys vs. Palmer/Chugiak, 4:45 p.m. (friendly match); Juneau girls vs. Chugiak, 6:30 p.m. JRC/The Alaska Club racquetball - After Tax Racquet Attack Party/Tourney: The entry deadline for this event is Wednesday, April 14, and the tournament takes place starting at 10 a.m. on Saturday, April 17. All players are guaranteed at least two matches in each division and they may play in a maximum of two events. Entry fee is $10 plus tax per event. All players must wear protective eyewear. A potluck dinner takes place after the tournament (see event flier for food assignments). Info: Eileen Wilson, 789-2181. Alaska Multiple Sclerosis Run/Walk - Meet at 9 a.m. at the Brotherhood Bridge Trail for a 10 a.m. event start. There is a 5-kilometer run and a 2-mile walk available. Info: Peggy Ann McConnochie, 586-3540. Sunday, April 18 Juneau Jumpers - Members of this competitive jump rope team will demonstrate their routines during a free show at 3 p.m. at the Glacier Valley Elementary School gym. The team is preparing for the U.S. World Team Qualifying Tournament, which takes place April 23-25 at Cleveland State University. Info: Scott Johnson, 789-6063. MEETINGS / CLINICS / ANNOUNCEMENTS Gastineau Channel Little League umpire training - Adult umpires are needed for the upcoming baseball and softball seasons. Umpires will officiate at various age levels, both behind the plate and on the bases. Flexible schedules exist with night and weekend games. The league season opens on Saturday, April 24. The last training clinic takes place from 10 a.m. to 4 p.m. on Saturday, April 17, at the Mendenhall River Community School. Info: GCLL Hotline, 789-9803. Juneau Alpine Club - Help build a new public-use cabin north of Bridget Cove by loading and unloading materials from a landing craft on April 17. Info: Jack Manning (of Territorial Sportsmen), 789-5540. Juneau Soccer Club U12 girls competitive team tryout - Tryouts for the JSC U12 girls competitive team (born Aug. 1, 1992, through July 31, 1993) will be held on Sunday, April 25, from 11 a.m. to 12:30 p.m. at Adair-Kennedy Memorial Park. The tryout fee is $25 per player with a signed JSC registration form. There is no fee for current JSC members. Registration forms are available at the tryout or online at http://www.juneausoccer.org. Tryouts for other teams were held earlier this month. Info: Noel Shima, 789-1525. OPEN GYMS / SCRIMMAGES / ONGOING Youth open gyms - Meet from 8-11 p.m. on Saturdays at Dzantik'i Heeni Middle School gym. Youth open gyms are free for students in grades 5-12. Info: Community schools director Joyce Kitka, 463-1700, Ext. 327. Adult volleyball/basketball open gyms - Meet from 7-9 p.m. on Fridays and from 7-9 p.m. on Sundays at Dzantik'i Heeni Middle School gym. Cost is $4 and open gym is limited to adults (age 18-older) only. Info: Community Schools, 463-1700, Ext. 325. Treadwell Arena programs - The ice arena is open seven days a week with a variety of public open skates, youth open skates, pond hockey and rental times available through Sunday, April 25 (see Treadwell Arena Web site for details). The rink is closed for ice maintenance from 7:45-10:45 a.m. on Mondays and from 7:45 a.m. to noon on Fridays. Fees for open skates are $4 adults, $3 youths (age 17-younger) and $2 seniors (age 65-older). Fees for pond hockey are $5 adults, $4 youths and $3 seniors. Skate rental is $2 for adults and youths, and $1.90 for seniors. Passes are available for open skates, pond hockey and skate rental. Facility rental is $150 an hour. Info: Treadwell Arena, 586-0410, or check on the Web at http://www.juneau.org/parksrec/icerink/. Adult indoor soccer open gyms - Sessions are held from 7-9 p.m. on Sundays and most Fridays at Floyd Dryden Middle School. Check http://www.juneausoccer.com for updated schedules. Open to players 18 and older; cost is $5 per session. Contact: Pat Race, 586-3440. Gastineau Channel Little League baseball open gyms - GCLL baseball open gyms for kids ages 8-12 will be held at Auke Bay School from 3-5 p.m. on Sundays beginning Feb. 21. Bring tennis shoes and a baseball mitt. Info: Lisa Baker, 789-4486. Juneau Department of Parks and Recreation Wednesday and Saturday hikes - For updates on locations of Wednesday and Saturday group hikes, call the Juneau Parks and Recreation Hikes Hotline. Always remember to bring plenty of food and water, wear warm layers of clothing and proper footwear for the hike. No dogs or firearms are allowed. Wednesday hikes are limited to hikers 18 years of age or older. Saturday hikes are open to families, and children must be accompanied by an adult. Info: Juneau Department of Parks and Recreation Hikes Hotline, 586-0428. Sports in Juneau is a service provided by the Juneau Empire to provide information on upcoming sports and outdoors events in Juneau. It runs when space is available. To submit an item to Sports in Juneau, please fax the info to 1-907-586-3028 or e-mail it to [email protected]. © 2017. All Rights Reserved. \| Contact Us
<urn:uuid:ef59e65c-e166-4bcc-be7a-165659acb49e>	New Travel agency opens: Cyndi Isaak, formerly of Southeast Executravel, has opened her own business, Cyndi's Cruise and Travel. The full-service agency focuses on vacation travel, cruising, groups and business travel. It is based out of her home, 415 Fifth St. in Douglas, 364-3435. Two other local agents who work with the agency are Carrie Graham at 789-6183 and Cassie Ritter at 789-9764. The Web site is www.travelgalileo.com/cyndicruiseandtravel. Eye doctors change location: Drs. Michael Bennett and Roy Box of Lifetime Eyecare moved next door to their old office last month. On May 22, they held a grand opening to celebrate moving into a new facility at 9309 Glacier Highway, Suite A 103. "Our practice is growing to meet the needs of the area residents in all areas of vision health services," Bennett said in a press release. "This move is part of that growth." Firm promotes accountant: USKH, an Alaska architectural, land surveying, planning and multidisciplinary engineering firm, recently promoted Samantha Emmal to the position of accounting manager. Emmal is based at USKH's Anchorage office, but works with clients in Juneau, Wasilla and Fairbanks. Emmal obtained her bachelor's degree in accounting from the University of Alaska Anchorage and joined USKH in 1996. Designer/project manager hired: Resource Data, an Anchorage-based Internet, database and GIS developer, has hired Matt Johnson as senior designer/project manager. Johnson is a nationally recognized graphic designer with 18 years experience whose work is in permanent collections of the Alaska State Museum, the Anchorage Museum, the University of Alaska Museum, and the Alaska Contemporary Art Bank. A former Juneau resident, Johnson's Juneau clients have included Sealaska Corp. and Perseverance Theatre. Bank names employment manager: Sue C. (Doggett) Johnson has been named a bank officer and promoted to employment and recruiting manager in First National Bank's Human Resources/Staffing Unit in Anchorage. Johnson, who joined First National in February 1999, will develop recruiting methods, supervise personnel interviewers and work on staff retention and career path plans. She will also develop relationships with community and professional organizations as well as government and private agencies. © 2017. All Rights Reserved. \| Contact Us
<urn:uuid:3947de10-154c-4d37-a653-0e38926f54db>	On the surface, it doesn't matter much to those of us in flyover country that Ben Silverman, the co-chairman of NBC Universal Entertainment, left the company this week after two years of abysmal last-place ratings. But Silverman's failure at NBC highlights the struggles networks are having connecting with audiences in a new age of media consumption. Network TV may not be the shared experience it once was, but it can be so much better than the current mess of blink-and-you-miss-it scripted shows lost in an ocean of reality TV. Nowhere is the problem more pronounced than at NBC, where the young and energetic Silverman took the creative helm just two years ago. As an entrepreneur, he had a record of success, developing the American version of "The Office," for example. But as a network suit, he was all over the place. A writer's strike didn't help, but almost half of the shows developed and launched during his tenure were of the reality ilk - stuff like "My Dad is Better Than Your Dad" and "American Gladiators." And some of his scripted shows were just retreads - "Knight Rider" and "Bionic Woman." The more fundamental issue, though, is the notoriously short leash on new shows nowadays. Of the first 23 shows launched under Silverman, 20 were canceled after 20 episodes or fewer (including 13 canceled after 10 or fewer). In many cases, that demonstrated mercy for the audience, but let's remember that "Cheers" and "Seinfeld" were early flops once, too. On cable channels, where ratings pressure is much lower, shows have time to grow and build an audience. Cable networks, including some owned by NBC, take advantage of the fact that more people can discover a show well into its lifespan through DVDs, DVR and digital downloads. All things are cyclical in entertainment. Remember when Bill Cosby was credited with saving the sitcom in the 1980s? Or when "Lost" was credited with saving drama just a few years ago? Well, for scripted shows to be saved, maybe they need a little more time to connect with people. That's quite a challenge when you hand one third of your primetime schedule over to Jay Leno. We understand the pressure from advertisers to produce results quickly, but viewers are tired, not just of bad concepts, but of seeing promising shows pulled away too soon. Sure, millions of Americans will watch a show called "Most Amazing Fishing Accidents" if that's what you dangle in front of them, but that leaves an empty spot in our pop culture soul - a spot that could be given to good writers who have time to find some traction with audiences. © 2017. All Rights Reserved. \| Contact Us
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<urn:uuid:2be2339c-3c50-4574-abf5-3d88510d5347>	Redmond, WA company Coros Wearables is launching a new helmet that comes with more than the usual bells and whistles. The product, launched on KickStarter, has been dubbed the Coros LINX Smart Cycling Helmet. In a nutshell, it is a bunch of useful technology stuck into a bicycle helmet. If you are going to wear a helmet, you might as well use one that comes with a smartphone app. Brooks England has added two new helmets to their iconic line of saddles, bags and clothing. These are currently being release in pre-order on their website, and are expected to ship by spring of 2017. Both helmets will be released in two sizes, M (53-58cm) and L (59-62cm). The ISLAND, our first Commuter Helmet, and the HARRIER, our first Road Helmet, are currently in development. Soon they will be in pre-production, the full commercial release being planned for Spring 2017 Adult bicycle helmet laws draw no end of passion on both sides. We don’t call them the “bicycle helmet wars” for nothing. But until recently I was under the mistaken belief that there were few – if any – jurisdictions in the United States that actually had laws regarding adult usage of bicycle helmets. This belief was shattered on a recent trip to Washington state where, coincidentally, two totally different people completely unconnected made comments about King County Washington having a helmet requirement for adults. I’ve ranted and raved about helmets and the odd fact that no helmet manufacture seems to advertise their testing strategies. I always found it odd that auto manufactures will show somewhat gruesome footage of crash test studies to show how safe their cars are, while helmet advertising never seems to even mention the word “safety.” Instead, we get sold $100 upgrades for improved cooling, better looks and lighter weight. Well, now those crazy Swedes have proven me wrong. Keep reading → For as many times as I’ve rambled, made fun of, attacked, or complained about helmets, you’d think I’d have come across this before. Ironically, it was a coworker that pointed this cultural phenomenon to me. So we take the worry about helmet hair and replace it with a hair helmet. That’s what you call turning a frown upside down. Of all of the things I’ve written about here on JustAnotherCyclist, few subjects tend to spark as much disagreement – from both cyclists and non-cyclists alike – as my posts regarding bicycle helmets. Many seeing me roll up to work or wherever without my helmet on have referred to me as reckless, stupid, crazy, nuts, or even… well, you can probably imagine. I’ve been told I’m an irresponsible parent, setting a bad example for my children. I’ve been told I make drivers on the road nervous, thus increasing motorists/cyclist contention. I’ve even been told I “deserve to crack my skull open” because I opt to sometimes ride without a helmet. I’ve never once encouraged anyone to ride without a helmet. Instead, I’ve spoke of my own opinions and ideas on the subject, encouraging others to find out the facts and make an educated, reasonable decision on their own. Cycling helmets seem to be a recurring theme here on JustAnotherCyclist as of late. To wear or not to wear. To promote or not to promote. Blah blah blah… There has got to be some way to resolve the issue. Perhaps with better technology? Folks seem pretty happy with the air bags in their cars. This probably has a lot to do with the reams and reams of evidence for their collective benefits. Unfortunately the same concept does not apply to bicycles. First off there is simply no practical way to mount an airbag on the handlebars of any but the most heavily customized bikes. More importantly is the fact that it is usually the ground or a car – and not the handlebars – that you smack into if you are injured in a cycling crash. If only there were some way to get that same level of passive protection on a bicycle… Maybe there is. I’ve been doing a fence-straddling maneuver here at JustAnotherCyclist regarding the issue of bicycle helmets. Again I will reiterate that I do not encourage folks to go about without a helmet. Nor do I encourage you to ride with one. It is entirely your choice (except for areas where specific laws apply.) My frustration comes entirely from the dogmatic nonsense that the issue seems to instill in some folks. I first started to become openly frustrated with the whole helmet situation when I was in the hospital for a broken collar bone due to a bike crash. I distinctly recall one of the ER nurses asking me “Were you wearing a helmet?” I simply answered the question at first, but then I started to think a little more critically about the question. Keep reading → I may, however, be changing my mind. And you, dear reader, get to come along for the ride. So to stop skirting the issue, I’ll state my opinion, as it exists today: I don’t really mind wearing a helmet, but I really don’t think they do squat to protect me. The risks the helmet protect me from are the same risks I experience when walking down the street. I’m just as comfortable riding my bike without a helmet as I am walking across my living room without a helmet. And that is when the “Say what” and “this guy’s nuts” comments come on. “Clearly you’re safer with a helmet on. It’s obvious. Anyone who thinks otherwise is a moron,” is another possible retort to my sentiment. Ah bicycle helmets. The topic that I just can’t leave alone. While I try to remain non-judgmental to the choices of others, and personally can take it or leave it, I still remain decidedly against helmet laws.
<urn:uuid:95ace876-e4d3-4ad3-aefa-1db360b5888c>	Ahhh the GoPro – the little camera that could. I’ve talked here about this camera before, and some of its wonderful features (or potential enhancements under development.) That’s because it is a damn fine, action sports oriented camera. I’ve shown a small subset of my cycling life as seen through the time-lapse view it provides, and I even caught my own crash in high definition glory. Well the folks at GoPro have been busy, and have release a couple of enhancements to the cleverly minimalist design.
<urn:uuid:1c7a4106-8908-4496-9aae-f76efb434594>	I love to travel and I will take whichever means of transportation necessary. I’ll fly or take the train when I can, but I’ve always had a soft spot in my heart for a good long road trip. Planning a road trip with kids makes me all nostalgic for my own childhood. Growing up in the 80s, my family of five would pile in the station wagon every summer for the journey from Florida to Tennessee for a week in the mountains. It was a long drive but I loved it. There’s just something about being in the car with family that means vacation has already started. Now that I am a parent, of course there are more responsibilities to consider. Things like naps and bathroom breaks and meals on the road and ALL. THE. PACKING can make taking a road trip with kids feel a bit challenging. But I still love it. Our boys have been great traveling buddies even from an early age so we actually choose driving most of the time when it comes to family vacations. When I was little, my sisters and I brought activity books and coloring books and paper for drawing and music to listen to on our walk-mans. While today’s technology is more advanced, our kids still bring most of the same stuff. It just got an upgrade. Read on to find out some of our favorites! 9 Ways to Rock your Next Road Trip with Kids This post uses affiliate links. If you choose to shop with my links it won’t cost you extra and you’ll support this blog! Thanks for reading Both our boys are really in to drawing and writing. Now that Calvin is learning to read, he loves practicing his words and letters. The Boogie Board is coming with us on our road trip to Florida in a couple weeks and the kids are so excited. You draw right on the board using the included stylus and when you want to start over, just push the button and it disappears! You can grab a Boogie Board on Amazon, but if you are a Sam’s Club member, that’s where I found the best deal for ours! We already have a van full of audio books for driving around town but I decided to round out our collection with Empire Strikes Back and Return of the Jedi. I love audio books because kids can flip through the pages and read or just listen if they are prone to car-sickness. Plus they are pretty inexpensive at around $5 each. Our van doesn’t have a built-in DVD system but we do have a portable one that we reserve for the long drives to Florida and Alabama. We have a case of favorite movies and this year we’re bringing Harry Potter along for the fun! The boys have seen the first couple movies so we’ll stick to those for now. I love having them all though for when they are a little older! I made this Mr. Potato Head Play Mat a few years ago and he will be traveling again this time. Since we are heading to Walt Disney World, I think he needs a few new accessories for the trip! Snacks and Water Any seasoned traveling parent will tell you snacks and drinks are essential for road trip success. We pretty much have an anything goes attitude about snacking on the road so we try to bring a few healthy options like fruit, pretzels and applesauce along with plenty of water. Letting the kids keep track of their own water is awesome too. We love the Pogo water bottles from Target when we’re on the go. Getting comfortable in the car can be difficult so we make sure to have lots of cozy items to keep complaints at bay. These fuzzy throws from Target are perfect for traveling. They aren’t too bulky but are super soft and cozy. Plus it’s fun to let the kids have something with their favorite characters on it. Why not? We never leave home without Puppy and Quackers. Everyone needs a cute little travel buddy, right? We don’t let the boys bring their whole room but a couple Beanie Boos usually find their way into the van as well. Activities (FREE Printable) A road trip with kids just wouldn’t be the same without a few car games! Growing up, we got a special snack each time we crossed a state line, searched for license plates and played the alphabet game. This year, I created a free Alphabet I Spy printable to have kids find words beginning with each letter and write them on the lines. Feel free to print more than one for a few rounds on your next trip. Want to add a challenge? No repeat words! Click image to open and print! For another fun activity, we just discovered the Dude Diary series at the school book fair and it’s already a huge hit. This journal for kids has fun would you rather questions, fill-in-the-blank stories and other imagination boosting activities. If there are any empty pages left by the time we leave, I’m sure this book will be coming with us. What are your go-to tips or activities for taking a road trip with kids? Share them in the comments below! Save this post for later or share with your friends! Thanks for reading and happy road-tripping!
<urn:uuid:2008ac9a-67d7-4d69-a150-362b945b2188>	The close up of Abraham Darby on the left shows the rich colours seen in this rose in New Zealand, while the photo on the right is taken in Reno with its more extreme temperatures. Is it more colorful in cooler climates, or is this variation a seasonal thing? Christine? Christine notes from Reno: 1. My first plant was purchased at the very tail end of a selling season so was a weak plant. 2. It is planted in a large pot in a less than optimal spot where it really gets too much shade. I have never felt confident that this plant is showing me what Abraham Darby can do, so I bought and planted another in summer 2007. There is now a nice plant growing in full sun, decent soil, and a good plant to start with. I will be observing it this coming season and will know more. 2008 UPDATE FROM CHRISTINE. A year ago I planted a new plant of Abraham Darby in good soil, in a sun-blessed location. Though I had grown this plant for many years in a too-shady spot, it had always been a weak grower, never recovering properly from a poor start. Since so many people think highly of this rose, I felt it deserved another chance in my garden. A year later, I can say without reservations that Abraham Darby is an astonishingly good rose in my climate! It is currently five feet wide and three and a half feet tall, and is simply loaded with bloom. The canes are very well branched with every branch tip having at least one bloom, with some bearing up to five. The blooms are definitely more pink than apricot but the fully opened blooms have an apricot cast in the inner petals, allowing the plant to blend beautifully with true apricot roses. The fact that Abraham Darby truly needs huge doses of sunlight is demonstrated by the fact that despite having a full, even circle of canes, every single bloom faces the direction of the strong, hot afternoon sun. Noting this, I realize that my too-shaded plant just never had a chance. In one year, this new plant has absolutely proven the worth of this variety. No wonder so many people love it, and now, I roses home \| garden pictures \| pink roses \| orange roses \| more David Austin roses See here for information regarding use of these Abraham Darby rose pictures. Copyright, JustOurPictures.com. Stock Photography
<urn:uuid:f57df2d5-f9f5-49db-a909-92da879a8bb7>	I’m sitting at a table with Walter Kresnik and Quintin Rodriguez. They’re bent over big sheets of paper with pencils and markers and paint, drawing hearts. We’re not talking about the kind of puffy, stylized hearts most at home on Valentine cards or sailors’ arms. These are the real deal — anatomically correct hearts, complete with valves and chambers and … veins. The room’s an open space the size of several classrooms put together, full of color and light and activity. Artwork covers the walls, and hangs from clotheslines, strung above tables full of working artists. Kevin Roach is one of them. “We start early in the morning, and then we're really creative and we think of something we want to draw and we don't copy. And it gives us joy!” he tells me. Roach has been making art here for almost ten years. Right now, he’s working on an incredibly intricate drawing of ants. Lots of them. Each one is outlined in black Sharpie, and then colored in with red. It looks like a lot of work. Roach says he’s been working on it for three weeks. “We don't rush anything. We like to keep things going but make it kind of beautiful.” "Why ants," I ask him? “Because they're real and they're a part of our world,” he says, and then bends back over his work. Real and part of our world. That’s a pretty good description of what it feels like to be here, and what Creativity Explored is all about. Visitors are welcomed. The doors to the studio are wide open. “The really, really important thing in all of our lives is human connection,” says Amy Taub, the executive director at Creativity Explored. “It’s that very simple moment when you look into somebody's eyes and you say hello. And it happens on a daily basis here.” Taub says the artists here come from all walks of life. Some have intellectual disabilities, some are on the autism spectrum, some have cerebral palsy. But here, they’re not defined by their disability -- they’re artists whose work is collected and exhibited and sold. “In the Bay Area there's been a real value placed on people's creativity and ways to communicate and that stems from the strong belief that all people have something to say,” says Taub. One of the teachers here is Victor Cartagena. He’s a Salvadoran-born artist who’s been working in the Bay Area since the 1980s. It’s his table where the drawings of ants and hearts are happening. “I teach what I do, the way I do my work, and that's the way I can guide them. Just introduce them to a new way of seeing that’s all about simplicity and lines. A way of thinking,” he explains. Cartagena leans in to look at Walter Kresnik’s anatomically correct hearts, and gives him a big smile. “Walter tends to do repetition, but every single heart is a totally different shape and size. But it's all coming from the same idea,” says Cartagena. The hearts represent people who are special to Kresnik. Cartagena asks him who they are, and carefully starts to write the names next to the hearts. At the other end of the table, Kevin Roach takes a break from his drawing, now covered with even more ants. “Are you ever surprised at the art you make?” I ask him. “Oh my God! I am. I'm really shocked. Sometimes I'm shocked that I even did it,” he says. And that’s when I notice what’s at the center of each ant: a little heart. To listen to this story, please click on the audio player above. If you would like to meet some of the artists and see their work, head over to Creativity Explored. Their new exhibit, Shoerageous, runs from Aug. 14 - Oct. 1, 2014.
<urn:uuid:9abe8c56-6321-4424-8ac7-7ba6d2d98121>	New Research Shows “Kangaroo Mother Care” Reduces Newborn Deaths More than 50 Percent, Proven to be More Effective than Incubators for Stable Preterm Babies. A new meta-analysis led by Dr. Joy Lawn of Save the Children, and published in International Journal of Epidemiology found a 51 percent reduction in newborn mortality when stabilized babies weighing less than four pounds (2,000 gm) received warmth and breast milk through continuous skin-to-skin contact on the chest of their mothers – kangaroo care. This makes it one of the most highly effective ways to give more babies the chance to survive and thrive. Not only is it simple and low cost, vital in less affluent counties but it also it is incredibly empowering for new parents making it equally important in more affluent countries too. A recent documentary Invisible Lives show its benefits in Malawi where it is widely practiced.
<urn:uuid:f6a24df9-0bbb-4f7f-9796-51c355a5d4a2>	I was thinking a lot about my girlfriends on Friday night, and how nice it is to have people out there who love you, and who tell you they love you, besides your mom and your grandma and your boyfriend. (And your dad and your brothers and your all your crazy aunts… I don’t want anyone to feel left out). Trevor’s been away for a week and I won’t see him until the end of the next one, so I was missing him a little and remembering the feeling I had for so much of sophomore year – part loneliness, part boredom – and being so glad I’d moved past that phase, when Steph called, just to see how I was doing. And we chatted for a little bit, and then we both had to go, and so we said our goodbyes, followed by quick “I love you”s. And it’s really nice, hearing that, in a sincere way, from someone who is far away but still cares. Later, I talked to Becca, and the same thing happened. And then I was less lonely, and happier. I also got some not-so-great news about another girlfriend on Friday night, someone who I love very much. It’s the sort of thing where there’s not much you can do other than wait and be supportive and try your best to understand. And let them know they’re loved. So this weekend, I’ve been thinking about her and sending love her way. And to all my other girlfriends, past and present. Even though that might be the hippy-dippy-est sentence I’ve ever written, that’s what I’m doing. Point being, go remind someone that you love them. In other news, I’m headed to Spain for work until Wednesday. It might be unlikely, but I’m hoping to get a few touristy hours in – any suggestions for the best thing to do in Madrid with 3 hours? And I’m bringing a few of these cookies with me, which might just be the best cookies I’ve ever made. Coconut, toffee, AND chocolate chips? Yes, please! Toffee-Coconut Crack Cookies Recipe adapted slightly from Two Peas and Their Pod. Makes 15 large cookies. These might be the most delicious cookies I’ve ever eaten. They’re super sweet, with a chewy-crunchy goodness from the melty toffee and toasted coconut. The dough is a little bit dry when you put them on the pan, but they hold together really nicely, forming some of the prettiest cookies I’ve seen. These are a must-try. - 1 c. shredded sweetened coconut (use unsweetened if you don’t love super-sweet cookies) - 1 c. flour - 1/2 tsp baking powder - 1/4 tsp baking soda - 3/4 c. brown sugar - 4 TBS (1/2 stick) salted butter at room temperature - 1 tsp vanilla - 1 egg - 3/4 c. Heath toffee bits - 3/4 c. chocolate chips - Preheat the oven to 350°F. Place coconut on a baking sheet in a single layer, and toast in the preheated oven for 5 minutes, until some pieces are golden brown. Stir together and set aside in a bowl. - In a medium bowl, whisk together flour, baking powder, and baking soda. Set aside. - In a large bowl, cream together softened butter and brown sugar. Add egg and vanilla, and beat until smooth. Mix in dry ingredients, then mix in toasted coconut, toffee bits, and chocolate chips. Form into disks with your hands and place on baking sheet. Bake for 12-15 minutes, until golden brown on the edges and beginning to crack on the top.
<urn:uuid:ee17b7bf-c27a-44f8-8d01-85697efe0a84>	Updated with corrections to story WESTPORT, Wash. – A man on a motorcycle with no lights on was killed on the Westport Highway last night. The Washington State Patrol has notified the family of 49 year old Kevin Crnich of Westport. He was struck when a car turned left in front of him on State Route 105 South of Twin Harbors State Park. A Northbound 1993 Mercury pinned the 1978 Yamaha on the southbound shoulder. A 35 year old McCleary woman who was a passenger on the bike was airlifted to Harborview Medical Center. The driver, an 81 year old Yelm woman was transported to Community Hospital in Aberdeen. Both riders were wearing a helmet, the highway was closed for several hours last night while State Patrol investigated the incident, both a cause and any charges are under investigation.
<urn:uuid:fb3bfac0-17b4-4047-a0d0-ab80f78a6686>	What’s a cozy mystery without tea and cookies? With stories in the news lately about hackers from China breaking into corporate computers and stealing proprietary software and information (for example, this 60 Minutes’ feature: The Great Brain Robbery), here’s a little historical gem about Robert Fortune (1812-1880) who accomplished the reverse. The low-tech version. From 1845-1848, this Scottish botanist managed to acquire China’s closely-guarded tea-growing and production secrets, along with actual plants to transplant in India. For the tea-drinking world, this was a game-changer. Within a short time, China no longer had a monopoly on tea, and Brits had control over the production of their favorite beverage. Robert Fortune was working for the Horticultural Society of London and had already traveled to China and learned a great deal about tea production, along with some surprises: Chinese merchants had been telling their customers for decades that green and black teas came from different plant varieties. Fortune learned that the difference between black and green teas wasn’t the variety of plant, but the method of drying the leaves. He also discovered that the Chinese were dyeing the green tea purchased by the English. He published his discoveries in book entitled Three Years’ Wanderings in the Provinces of China. It drew the attention of the East India Company, which commissioned him to return to China and acquire tea plants for them. In Fortune’s own words: I was deputed by the Honourable the Court of the Directors (sic) to proceed to China for the purpose of obtaining the finest varieties of the Tea-plant, as well as native manufacturers and implements, for the Government Tea plantations in the Himalayas. Fortune had no ethical problem with such a request. In his view, plants belonged to the world for everyone’s use. The Chinese were incredibly secretive (and rightly so, given what did happen) about how their tea was produced. Fortune spent two and half years in China, shaving his head and adopting the attire of a Chinese merchant (read Sarah Rose’s fascinating book, listed below, for more details). It was sometimes a challenge to evade China’s in-port restrictions, which only allowed foreigners to travel one day’s distance from the ports allowed to Europeans by treaty. But Fortune managed to travel to areas few Europeans ever saw. Here is Fortune’s own account of how he transported the tea plants he collected, using what was called a glazed case, first devised by Nathaniel Bagshaw Ward: The mission was a rousing success: Upwards of twenty thousand tea-plants, eight first-rate manufacturers, and a large supply of implements were procured from the finest tea-districts of China, and conveyed in safety to the Himalayas. Wow, 20,00! And the equipment and experts to successfully start a tea-growing operation in India. Can you imagine? Want to read more? Fortune, Robert. Three Years Wanderings in the Provinces of China. London: Spottiswoode and Shaw, 1847. Fortune, Robert. A Journey to the Tea Countries of China; Including Sung-Lo and the Bohea Hills. London: W. Clowes and Sons, 1852. Rose, Sarah. For All the Tea in China: How England Stole the World’s Favorite Drink and Changed History. New York: Viking Press, 2010. Click here for Amazon link. Happy tea drinking, P.S. – Be sure to check out the following post for my giveaway and flash fiction fun! Entry deadline is January 31.

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