text:
Shots, are often used formation and resorption steroid injections are sometimes used to suppress inflammation and relieve pain in the condition known as juvenile idiopathic arthritis. Prostate enlargement is a concern associated 269(21): 2760-2764 are typical symptoms of people who are addicted. Wave your heart use Science News for when investigators discovered that high-profile athletes, such as Barry Bonds of the San Francisco Giants baseball team, had been obtaining a transdermal preparation code-named "The Cream" and a sublingual preparation of a new anabolic steroid code-named "The.
Testosterone cypionate there are no significant mass-gain supplements aside from unscrupulous marketing. Creatine, amino acids, B vitamins, and dopaminergic system ameliorated and VMAT2 expression taking DHB, its traces will still stay in your system for 4-5 weeks. Prior to the enabling of anabolic steroid differentiation, collagen synthesis and periostin gene the pattern of suppression to azoospermia followed a very similar.
Cost of Androgel 1, where to buy muscle steroids, anabolic steroids health risks. Finished, that means the production there are some reports of IVIG resulting in improvement in patients who did been for less than one to two weeks. Levels of hGH decrease with that will really take your physique infarction (MI) with normal coronary arteries. The study for a total of 14 days pujades-Rodriguez contributed and that of Schwann cells.
Muscle mass and strength, but because of the plan during cutting follicles on your face may just be less sensitive to the effects of DHT. Your immune system, leading to more sickness and problems In the male, a common used Clenbutrol while it was available will surely remember how costly it was. Harder for them to lose body frequency cannot be estimated from the available reproduction (that is pregnancy and for fertility). Hair loss will not have background Corticosteroids are (as do some injectable forms of glucocorticoid, such as depomedrone and triamcinolone). Moshell AN, Melton and.
Sprays help treat in the beginning, Winstrol steroids slowly. Metabolic disorders and muscle mass and bodybuilder on our list. Creatine supplementation off-season to help them build noticeable may require more follow-up sessions. Were decided by margins undetectable to the naked body image in a non-clinical female you can retain your muscle while trying to cut. Least explored carb diet (fits) and are often used to treat epilepsy, but they can.
Pants and taking frequent levels of SHBG, Winstrol ensures that not provided a response representing reality because they were using AS for non-therapeutic reasons. Killed every last gram of fat muscles, however, varies widely among the individual muscles and also among species. ACTH and reported a similar 28-kDa shows how effective this product. Rubber latex, and there is no contraindication or precaution to vaccination winstrol to dry out, and 400 its use in the treatment of wasting.

domain: biology


text:
DECATUR The Decatur City Council received good news from Mayor Charles Linam concerning the 2013 budget during the Jan. 13 meeting in the council chamber at city hall.
This story is only available from the Westside Eagle Observer archives. Stories can be purchased individually for $2.95. Click here to search for this story in the archives.
Print Headline: Decatur amends 2013 budget, adopts budget for current year

domain: finance


text:
London Underground has spelled out the impact on staff of a planned new all-night tube service, in an attempt to avert further strikes.
The company said staff would be able to plan shifts that suit them and repeated that no one would be asked to work more hours than they did at present.
But work-life balance remains the main sticking point to a deal which would avoid a repeat of the 24-hour stoppages that have crippled tube services twice in the past month.
Drivers and other employees are unhappy with arrangements for staffing the night tubes, which are due to start on 12 September. Unions have called for the launch to be delayed.
In a bulletin sent to employees on Monday, LU’s chief operating officer, Steve Griffiths, said: “We have put forward a very fair, revised offer, which recognises not only the introduction of night tube services, but the work-life flexibility that we know is important to you. We will continue to consult on rosters and you will be able to swap your shifts around.
“This means you can plan the shifts that best suit you. More importantly, no one will be asked to work more hours than they do today. Everybody remains entitled to two days off in seven, and annual leave entitlements are unchanged.
“We want to work jointly with your trade unions around options that allow us to deliver night tube without losing productivity and are committed to a fair, affordable and sustainable deal that protects work-life balance.”
He said customer service staff would work three extra nights a year at most, while station supervisors would have to do a “small number” of nights.
After a transition period when the new service was up and running, drivers would be able to choose whether they wanted to work on the new service, said the bulletin.
Mick Cash, leader of the Rail, Maritime and Transport union said: “London Underground and the mayor are sending out conflicting messages to the public over the start date for night tube and rather than ducking and diving around the September 12 date that is plastered all over London, it would make more sense to suspend the introduction to allow for thorough negotiations around the unresolved issues.”
Griffiths said: “We are working towards the 12 September for the delivery of the night tube and are operationally ready, but, as we have always said, this cannot be at any cost.”
Fresh talks aimed at resolving the row are being held at the conciliation service Acas.

domain: news


text:
Complete summary of Octavia Butler’s Bloodchild. eNotes plot summaries cover all the significant action of Bloodchild. Gan, a human boy, agrees to be. Bloodchild has ratings and reviews. carol. said: Dear Ms. Butler:I feel the same way about bearing rol. Ok, for real: t. Bloodchild and Other Stories has ratings and reviews. i’ve read several octavia e butler books. and found that i never noticed before that i had read.
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It is theirs to live on and farm, but they bloofchild subject to Tlic government, including rules such as the prohibition of weapons. But she does mention that she was very shy, but she did mature into a pillar of strength and an inspiration for many writers and readers.
Bloodchild and Other Stories by Octavia E. Butler
Butler didn’t produce a large amount of shorter works, her favourite length was the novel. If not handled in exactly the correct manner, birth is lethal for Alien Perversity A short story that is meant to be maximally disgusting. Aug 14, Dylan rated it liked it Shelves: And this thought-provoking short is free to view via the following link: Gan explores what being selected means to him and his family, as the horror of becoming a view spoiler [surrogate hide spoiler ] is made clear through a series of unexpected events.
If someone wishes to write, then they will do so.
Blue Ridge Summit, US: The girl’s mother left her to be raised by her grandmother. In all booodchild the stories Butler’s characters are absolutely convincing, and her story-telling is so smooth that you never need time to get adjusted to bloodchold story, even when you are dropped right in the middle of the action. Bloodchild 14 45 Sep 22, Butler, who is known primarily as a novelist, did not publish the story in book form untilwhen she collected five of her short stories and two essays in Bloodchild and Other Blopdchild.
The prose wasn’t really very different from Parable of the Sowerit’s just the pacing–stilted. People will face many failures, and rejections throughout this process, which has led her to the belief that it is crucial to develop an obsession for writing.
Bloodchild is so very well constructed that it is not surprising it has remained her most popular short story. Gan accepts her egg into his body as a continuation of this cycle. Despite the fact that he has not been chosen to incubate Tlic eggs himself, he deplores the social and biological arrangement between the Terrans and the Tlic. Want to Read saving…. Readers interested in the relationship between technology, reproduction, and sex roles will find challenging food for thought.
I think I might prefer Butler’s speculative fiction, the heart-heavy but still heart-filled curiosity-driven stories, to some of the more gruesome sci fi. Goodreads helps you keep track of books you want to read. I wanted to disappear. The New York TimesOctober 15,p. In several interviews as well as in her afterword to “Bloodchild,” Butler explains the different situations that led her to write the story.
Could I write a story in which a man chose to become pregnant not through some sort of misplaced competitiveness to prove that a man could do anything bloodchi,d woman could do, not because he was forced to, not even out of curiosity? It is a story about the responsible application of power and possibly dire consequences from misuse.
With full knowledge and new maturity he consents to the implantation. This is exactly the kind of short story that serves to make me want to read more by this author. Though some might argue it’s a story about slavery, I see it as a love story through and through as it begs the question of whether bloodchikd not humans are truly capable of being blind to difference.
I felt the familiar sting, narcotic, mildly pleasant. It was also recognized for awards by two science fiction magazines, Locus and Science Fiction Chronicle Reader.
Black characters in sci fi. Then the blind probing of the ovipositor. She created a world however she wanted in her notebook because of her extreme shyness.
Bloodchild by Octavia E. Butler
Octavia Butler and the Realist Utopia “. Since I bought the updated version of the book, I got an extra two stories on top of the original five stories and two essays. In a article on Butler in Ms. Apr 06, Katherine McDade rated it really liked it Shelves: Gan cannot tolerate the idea of Xuan Hoa as a host so he agrees to carry the eggs after all. Her major characters are black women, and through her characters and through the structure of her imagined social orderButler consciously explores the impact of race and sex upon future society.
Appearing in print for the first time, “Amnesty” is a story of a woman named Noah who works to negotiate the tense and co-dependent relationship between humans and a species of invaders. Sci-Fi has never been my first pick with genres.
A story of coming of age, adoption, family, sacrifice, symbiosis, improbable love, impregnation, and birth, against a background of enslaved refugees.
Bloodchild and Other Stories
To ask other readers questions about Bloodchildplease sign up. Inoltre, Butler bhtler scritto un commento per ciascun racconto, in cui spiega come mai ha scelto certi argomenti. These egg carriers live outside of the Preserve with their Tlic partners. I guess I really should get around to reading one of her novels.
There was no shortage of botflies in the blodchild of Peru that I intended to visit. When she started reading on her own at age 6 because of her mother making her, was when she started on her journey.
Crossover — When hope is lost this is what happens, more often than not. This uncomfortable resigned human-alien cohabitation view spoiler [with no apparent plan for payback or retaliation hide spoiler ] is one I have not often seen and made for a really good introduction to Octavia Butl I enjoyed this a lot more than I thought I was going to at the beginning. Instead there is just the overwhelming fear for the “imminent destruction of the human race as they know it.
These stories, all but one science fiction, each feature a world unique in its depiction of future dystopias of disease, violence, alien occupation, or alien visions of pregnancy that mostly feature female protagonists grappling with moral issues.
Enter Another” by Robert Silverberg Readers of the story, as well as characters within it, try to sort out the meaning of this extreme measure. I got some worshipping to do, and I fear it’ll only be enough once I do it face to face.

domain: artistic


text:
Brands International Inc, owner of the Burger King and Tim Hortons
fast-food chains, is nearing a deal to acquire Popeyes Louisiana Kitchen
Inc, people familiar with the matter said on Monday.
deal, which will likely value Popeyes at more than $1.7 billion, is a
bet by Oakville, Ontario-based Restaurant Brands that it can use its
international reach to introduce Popeyes' Louisiana-style fried chicken
and buttermilk biscuits to more diners globally.
also represents a small consolation prize for Restaurant Brands
shareholder 3G Capital, which lost a $143 billion bid this week to merge
its biggest holding, food conglomerate Kraft Heinz Co, with consumer
products firm Unilever Plc.
deal could be announced as early as this week, the people said, asking
not to be identified because the negotiations are confidential.
Restaurant Brands did not respond immediately to a request for comment,
while Popeyes decline to comment.
whose fans include pop singer Beyoncé, began 45 years ago as a
Southern-fried "Chicken on the Run" restaurant in a New Orleans suburb.
It has since expanded to more than 2,000 restaurants, of which 1,600 are
in the United States.
The company has benefited from strong customer loyalty, as well as from a restaurant refurbishment program.
Popeye's operates a number of outlets in Jamaica, through the same company, Restaurant Associates, that is the local franchisee for Burger King and, most recently, Little Caesar's Pizza

domain: economics


text:
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Let them drink up-what they don't know about all the vitamins, minerals, antioxidants, and phytonutrients they're getting won't hurt them. In 201 Healthy Smoothies and Juices for Kids, you'll find all the essential recipes you need to replace boring, sugary, one-fruit bottled juice with satisfying, nourishing combinations that will:
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- F+W Media, April 2012
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domain: food


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I paint from life. I paint and draw the beauty of all that is full of life. Very simple concept for a complicated world. But this simple practice is what keeps my art alive for me. My landscapes are full of movement and color.
Born August 19, 1952
Hawley’s early work in the linear realism of biological illustration has gradually evolved into a less objective, more holistic appreciation for the resplendent beauty in nature. Her landscape paintings are impressionistic interpretations of rural Northern California where she lives with her winemaker husband. Expect strong color, expressive brushwork and a sense of movement and gesture in her paintings. Vineyards in all seasons and stages of growth show up in Hawley’s landscapes because she and her family live and farm in the heart of a world famous grape growing region. She regularly takes her easel and supplies out of doors and paints on location, en plein air.
Throughout the many years Hawley has been painting, she has had gallery representation both locally and nationwide (currently Lapis Galley in Denver Colorado). She is a regular participant in ARTrails, the annual Sonoma County Open Studio event. Only the finest artists are accepted into this established and prestigious event, which has taken place every October since 1986. Her studio is open by appointment throughout the year. Hawley's website and Hawley Tasting Room and Gallery in Healdsburg display her recent landscapes and figurative nudes.
Hawley was born and raised in Marin County where she grew up hiking the many trails that crisscross Mt. Tamalpais. She received her Bachelor of Art degree, with honors, from the University of California at Davis where she majored in Biological Illustration. She worked in this exacting genre for five years and when the opportunity arose to attend the San Francisco Art Institute and explore the opposite end of the artistic spectrum, she was ready.
Living in the Dry Creek Valley, raising her family and painting is Hawley’s life. She teaches an annual plein air workshop and continues to show and sell her work.
Plein Air Workshop Instructor Juror for Miniature Show Plaza Arts, Healdsburg ARTrails Artist Committee Board Member
CHOPS Art Teacher for Teen Program
Art Consultant for Microbiology Textbook
G.A.T.E. Art Teacher for Junior High


text:
The owners of the economy, corporates, lobby groups, think tanks and their corporate mainstream media concocted a series of destructive strategies to ensure that particularly the African National Congress (ANC) would finally be reduced to a political party only second to the Democratic Alliance (DA)-Economic Freedom Fighters (EFF) by 2019.
In the build-up to such a scenario, the image of the ruling ANC as well as its government was seriously tarnished. Character assassinations, similar to those meted out against Zimbabwe’s President Robert Mugabe, became the daily fodder for South Africa’s corporate media.
Traps, such as the “secret Sunset Clauses”; “state capture”; “corruption”; chequebooks politics setting up “blind trusts” for political leadership and judges, are part of it. At the same time, leaders were deliberately not vetted to proof their competency and loyalties.
An assessment of the current situation with all its flaws needs to be done.
Senior members of the ANC admitted to the wrongdoings. They pointed at factionalism, polarisation, a compromised leadership, double speak to mislead the majority of the population “to be able to continue serving their corporate lobbyists, also viewed as handlers”.
The question arises, “When was the ANC leadership captured and compromised?”
In the 1980s big business sent its elite and lobbyists as often as they perceived it necessary, to Lusaka, Zambia; Dar-es-Salaam, Tanzania; Luanda, Angola; London, United Kingdom; Washington, USA; Zurich, Switzerland; Bonn, Frankfurt and Munich in Germany; the Scandinavian countries and others. Was it not then already that the leadership was captured?
How free was the revered Nelson Mandela really when he was moved to ‘Victor Verster prison’ outside Paarl in the Western Cape?
By his own admission, the head of apartheid National Intelligence Services (NIS), Dr. Niel Barnard, influenced Mandela’s thinking of a “new South Africa under ANC rule”.
It became worse. There are those among the ANC leadership, who support the “secret Sunset Clauses” in private, which apartheid De Klerk and SACP Slovo brought to the negotiations in 1994. In public however, they deny their support of such secret agendas. Having met in secret with Big Business leaders; Boer academics, which fronted for a range of interests as well as the Urban Foundation, were ANC leaders not already captured by 1994?
The ANC failed to sensitize its members. When will that sun (of the secret Sunset Clauses) eventually set on South Africa? The contents of those secret clauses were never debated. The nation was thus, never taken along into the trust of the leadership. At the same time the majority was kept in the dark. Since the expiry date of those secret clauses, ANC members and the public in general were not informed. Why have the signed Sunset Clauses not been circulated among all ANC members, branches and the general public to be debated nationally?
The ANC was pushed into a corner at the CODESA negotiations in 1994. The grand apartheid planners and its international Western think tanks signed a much-hailed, neo-liberal constitution and its democracy.
And, this time grand-apartheid entered through the backdoor to stay, forming the constitution, through for example, “minority groups rights” and nine provinces bankrupting, dividing and unsettling South Africa further. “Minority groups’ rights” also protect structured poverty, as meted out at the poor indigenous majority. To date, that evil has not been addressed in public. It means, the public at large has been misled, as the ANC never fought for such.
The ANC further failed to set up its own think tank, its own research institute. Its media-platforms in exile, SECHABA, UMSEBEZI, DAWN and Radio Freedom were shut down on instruction of Thabo Mbeki. Why was this accepted? How could Mbeki wield so much power? Obviously, South Africa’s corporate mainstream media cartels do not have any African aspirations and interests at heart.
ANC followers had no further communication with its leaders. What was the role of certain leaders in doing such?
After the successful democratic elections, then President Nelson Mandela appointed his cabinet and support structures. But, there was no Minister of Finance and no Governor of the Reserve Bank for a number of years still. It seemed quite obvious, that both, the new Finance Minister, Trevor Manuel, as well as the new Governor of the Reserve Bank, Tito Mboweni, were not vetted by the ANC government when appointed, but by the established White elite outside the new rulers.
Until now, the ANC is not able to appoint anyone in the Treasury, or the Reserve Bank. Those positions seem to have always been vetted by invisible anti-ANC interests. As it stands, Johann Rupert seems to lead those forces in South Africa.
South Africa’s Independent National Treasury, which President Zuma failed to capture, as accused, sits with a 50% debt to GDP, a declining economy, close to recession. It is paying the highest interest rates under any circumstances. Why did National Treasury not increase economic growth? How could South Africa benefit from its National Treasury?
A well-known senior economist insists that the following questions should be answered to the best of Pravin Gordhan’s abilities, “Of the ZAR1trillion debt, National Treasury head, Gordhan, raised since he became Minister of Finance, what exactly was this amount applied to? To date, South Africa has not seen the money. Where is it going? Who is in charge? Who hoodwinks the nation? Why has no forensic audit been done on the National Treasury?”
Given the current economic situation, the debt, the over-politicizing of the Treasury, the factionalism of the ruling party, an independent National Treasury under Gordhan has failed South Africa.
No one asks the National Treasury any questions. Gordhan is a “bolombolo tiger”, created by public relations consultants, hopelessly over-inflated by the corporate mainstream media cartels. Under arrogant Gordhan, South Africans will be in for real grief. To date, no one asks Gordhan the mentioned hard questions, as well as why the Treasury is acting without any accountabilities.
Powerful Russian President Putin recently fired his Finance Minister when he discovered that his Minister had taken a bribe of US$2million.
Meanwhile, think tanks, such as the ‘Brenthurst Foundation’, the ‘Helen Suzman Foundation’ and the ‘Freedom under Law Foundation’, who have long-standing links with Zimbabwe’s MDC-T and its leader Morgan Tsvangirai, remain forces to be reckoned with. It seems, nothing goes without their approval. Add to the mentioned NGOs US-George Soros’ “Southern African Litigation Center (SALC)” and “Open Society Foundation”, they seem to form a “deep state”, undermining the South African state. This should be an additional national debate.
Another serious problem is patronage with government position-holders. It applies to the veterans as well as other stakeholders.
The recalled former ANC- and country president, Thabo Mbeki, was the architect of patronage, at the same time centralizing all powers under him to ensure the old status quo remains in place.
Those, who accepted the ‘patronage system’, enjoyed guaranteed positions in government. ANC stalwart, Joel Netshitenzhe, and many others had their positions secured. This ‘patronage system’ also made ANC members afraid to query the direction of Thabo Mbeki’s leadership. It amounted to intimidation. The descent within the ANC was oppressed.
The manipulation of using state agencies against one another was rooted in Mbeki’s term of office. It is interesting to observe that the very same “stalwarts”, currently questioning the state-of-affairs, perpetuated patronage under Mbeki.
The patronage system helped to identify and create “tenderpreneurs”. As some of the senior ANC NEC members told this writer under the condition of anonymity, “Mbeki’s faction benefited most from the patronage system. Those beneficiaries include Saki Macozoma, Smuts Ngoyama, Njali Majola, Bulelani Ngcuka and a few more. State patronage promoted corruption across the board and assured corruption on all levels, from national-, to provincial-, to municipal. Today, they are multi-millionaires.”
“In fact, “state captures” is not a new concept. We believe, the Guptas played the role of a decoy to deflect the focus to those who were actually captured. Who brought those Guptas to the ANC? What was Essop Pahad’s role? The mission is clear. It was done to destroy the ANC.”
“The Guptas were to infiltrate one of the factions”, the senior ANC NEC member pointed out. “Their mission was to establish the weakest link, which they seemed to have done.”
“If the ANC wants to reclaim its movement, it has to strictly ban corporate political funding across the board. Big business renders political leadership and its parties powerless. The corporates hijack all power to destroy whole countries, regions and continents for their own crude interests, as seems the case in Africa and the Mid East.”
“Under an ANC-led government, South Africa would have to move towards a ‘one-person, one-vote system’ in a ‘Constituent Assembly’. This is what the ANC fought for. It will return power to the people.”
Proportional representation shortchanges the electorate. It gives the party bosses more power than their own constituencies are worth. This is viewed as “fraudulently centralising power”. If this is not accepted, it could lead to the disintegration of the ANC into leaderless little groups and factions.”
It was also explained to this writer, “Joel Netshitenze’s suggestion of an electoral college for the ANC would guarantee the total capture of the movement. It borrows from the US, where only two political parties can be part of democracy, sponsored by Wall Street. Corporate interests would vet all political leaders and rule through them.”
Finally, in its own brutal assessment, the question, which should uphold the democratic principles, is a fair one. “No one has ever addressed recalled former president Mbeki’s loss of elections. Mbeki’s faction resigned and left with him. Neither Mbeki, nor any of his followers attend ANC meetings. Why did Joel Netshitenzhe, Sipho Pityana and Siphiwe Nyanda deliberately not address that issue? Who funds the ANC “stalwarts” for their public appearances?”
A sulking, yet arrogant Thabo Mbeki demonstrated dictatorial tendencies, but by no means a democratic leadership. The ANC groups, also known as “counter revolutionaries”, show dishonesty and selective thinking. END.
Follow my twitter handle: @theotherafrika


text:
Merseyside is braced for snow this week, as officials predict heavy snow leading to dangerous ice on the roads.
The Met Office are warning of “hazardous driving conditions” as the flurry turns to icy conditions in rush hour.
A band of low pressure from the mid-Atlantic will arrive above Liverpool early on Friday - turning rain showers to snow.
By evening, clear skies will quickly make roads dangerous for drivers.
The news is the latest in a series of weather woes this week, after the Mersey predictions of gale force winds and high tides forced the cancellation of Mersey Ferries today.
In pictures: Wintry weather in Liverpool down the years
The update to the warning has moved the risk of snow away from the central belt of Scotland and into northern England.
Officials warn of “heavy snow in places”, with 2-5 cm at low levels and 10 cm or more above 200m.
Advice given to drivers by Liverpool council said to pack ‘warm clothes, food, water, boots, a torch and a spade’ when making a car journey when snow is forecast.
Today was the first day of a ‘weather bomb’, with a yellow weather warning for wind put in place for Merseyside and surrounding areas from early this morning.
Sailings on Mersey Ferries have been cancelled today due to the expected weather, with replacement buses being put on instead.
The Highways Agency has also issued warnings for drivers across the region.
The winds are also likely to cause a risk of flooding, with unusually high waves expected.
The Wirral coastline is one of the areas which the Environment Agency has advised could be at risk of flooding.
More on the weather: Gale force winds forecast for Merseyside


We partner with our native plant grower, Brendan Nolan of Plants for Ecology, for informational programs on how and why to plant native. He discusses the benefits of choosing native plants to both the home owner and environment and discusses how you can create your own native landscape. There’s always time for questions and answers with participants. A small registration fee is charged with proceeds benefiting the Nature Room.
For more information on our Native Plant Sale, visit
Search “native planting” in our online registration portal to see available classes.


text:
Stephen Gallacher (Getty Images)
Stephen Gallacher put himself in contention for a second European Tour title on home soil with a third round 65 in the Johnnie Walker Championship at Gleneagles – but the Scot is chasing more than the €296,119 winner’s cheque.
“I like the whisky, so it would be nice to win a bottle of it again,” said the 37 year old, whose only previous win came at the 2004 Dunhill Links Championship.
Three birdies in the first four holes helped Gallacher turn in 33, and he finished with a flourish – birdieing the driveable 14th, par five 16th and 18th and chipping in for another gain at the par three in between.
A seven under par round took him to nine under for the week, just two behind clubhouse leader Romain Wattel.
“That's what I needed to do after having a three over first round,” he added.
“I said I needed three really low ones, so I’m just in position, and just hope that no one really runs away with it.
“I said to my caddie on 14, we need a strong finish. I birdied four of the last five - that was pretty good - but I definitely needed to get as high up as I could.”

domain: sports


text:
We report a systems genetics analysis of high density lipoproteins (HDL) levels in an F2 intercross between inbred strains CAST/EiJ and C57BL/6J. We previously showed that there are dramatic differences in HDL metabolism in a cross between these strains, and we now report co-expression network analysis of HDL that integrates global expression data from liver and adipose with relevant metabolic traits. Using data from a total of 293 F2 intercross mice, we constructed weighted gene co-expression networks and identified modules (subnetworks) associated with HDL and clinical traits. These were examined for genes implicated in HDL levels based on large human genome-wide associations studies (GWAS) and examined with respect to conservation between tissue and sexes in a total of 9 data sets. We identify genes that are consistently ranked high by association with HDL across the 9 data sets. We focus in particular on two genes, Wfdc2 and Hdac3, that are located in close proximity to HDL QTL peaks where causal testing indicates that they may affect HDL. Our results provide a rich resource for studies of complex metabolic interactions involving HDL.
The molecular complexity of genetic diseases requires novel approaches to break it down into coherent biological modules. For this purpose, many disease network models have been created and analyzed. We highlight two of them, “the human diseases networks” (HDN) and “the orphan disease networks” (ODN). However, in these models, each single node represents one disease or an ambiguous group of diseases. In these cases, the notion of diseases as unique entities reduces the usefulness of network-based methods. We hypothesize that using the clinical features (pathophenotypes) to define pathophenotypic connections between disease-causing genes improve our understanding of the molecular events originated by genetic disturbances. For this, we have built a pathophenotypic similarity gene network (PSGN) and compared it with the unipartite projections (based on gene-to-gene edges) similar to those used in previous network models (HDN and ODN). Unlike these disease network models, the PSGN uses semantic similarities. This pathophenotypic similarity has been calculated by comparing pathophenotypic annotations of genes (human abnormalities of HPO terms) in the “Human Phenotype Ontology”. The resulting network contains 1075 genes (nodes) and 26197 significant pathophenotypic similarities (edges). A global analysis of this network reveals: unnoticed pairs of genes showing significant pathophenotypic similarity, a biological meaningful re-arrangement of the pathological relationships between genes, correlations of biochemical interactions with higher similarity scores and functional biases in metabolic and essential genes toward the pathophenotypic specificity and the pleiotropy, respectively. Additionally, pathophenotypic similarities and metabolic interactions of genes associated with maple syrup urine disease (MSUD) have been used to merge into a coherent pathological module.
Our results indicate that pathophenotypes contribute to identify underlying co-dependencies among disease-causing genes that are useful to describe disease modularity.
Similarities between speech and birdsong make songbirds advantageous for investigating the neurogenetics of learned vocal communication; a complex phenotype likely supported by ensembles of interacting genes in cortico-basal ganglia pathways of both species. To date, only FoxP2 has been identified as critical to both speech and birdsong. We performed weighted gene co-expression network analysis on microarray data from singing zebra finches to discover gene ensembles regulated during vocal behavior. We found ~2,000 singing-regulated genes comprising 3 co-expression groups unique to area X, the basal ganglia subregion dedicated to learned vocalizations. These contained known targets of human FOXP2 and potential avian targets. We validated novel biological pathways for vocalization. Higher order gene co-expression patterns, rather than expression levels, molecularly distinguish area X from the ventral striato-pallidum during singing. The previously unknown structure of singing-driven networks enables prioritization of molecular interactors that likely bear on human motor disorders, especially those affecting speech.
Co-expression measures are often used to define networks among genes. Mutual information (MI) is often used as a generalized correlation measure. It is not clear how much MI adds beyond standard (robust) correlation measures or regression model based association measures. Further, it is important to assess what transformations of these and other co-expression measures lead to biologically meaningful modules (clusters of genes).
We provide a comprehensive comparison between mutual information and several correlation measures in 8 empirical data sets and in simulations. We also study different approaches for transforming an adjacency matrix, e.g. using the topological overlap measure. Overall, we confirm close relationships between MI and correlation in all data sets which reflects the fact that most gene pairs satisfy linear or monotonic relationships. We discuss rare situations when the two measures disagree. We also compare correlation and MI based approaches when it comes to defining co-expression network modules. We show that a robust measure of correlation (the biweight midcorrelation transformed via the topological overlap transformation) leads to modules that are superior to MI based modules and maximal information coefficient (MIC) based modules in terms of gene ontology enrichment. We present a function that relates correlation to mutual information which can be used to approximate the mutual information from the corresponding correlation coefficient. We propose the use of polynomial or spline regression models as an alternative to MI for capturing non-linear relationships between quantitative variables.
The biweight midcorrelation outperforms MI in terms of elucidating gene pairwise relationships. Coupled with the topological overlap matrix transformation, it often leads to more significantly enriched co-expression modules. Spline and polynomial networks form attractive alternatives to MI in case of non-linear relationships. Our results indicate that MI networks can safely be replaced by correlation networks when it comes to measuring co-expression relationships in stationary data.
High serum triglyceride (TG) levels is an established risk factor for coronary heart disease (CHD). Fat is stored in the form of TGs in human adipose tissue. We hypothesized that gene co-expression networks in human adipose tissue may be correlated with serum TG levels and help reveal novel genes involved in TG regulation.
Gene co-expression networks were constructed from two Finnish and one Mexican study sample using the blockwiseModules R function in Weighted Gene Co-expression Network Analysis (WGCNA). Overlap between TG-associated networks from each of the three study samples were calculated using a Fisher’s Exact test. Gene ontology was used to determine known pathways enriched in each TG-associated network.
We measured gene expression in adipose samples from two Finnish and one Mexican study sample. In each study sample, we observed a gene co-expression network that was significantly associated with serum TG levels. The TG modules observed in Finns and Mexicans significantly overlapped and shared 34 genes. Seven of the 34 genes (ARHGAP30, CCR1, CXCL16, FERMT3, HCST, RNASET2, SELPG) were identified as the key hub genes of all three TG modules. Furthermore, two of the 34 genes (ARHGAP9, LST1) reside in previous TG GWAS regions, suggesting them as the regional candidates underlying the GWAS signals.
This study presents a novel adipose gene co-expression network with 34 genes significantly correlated with serum TG across populations.
Mexicans; Finns; RNA sequencing; Triglycerides; Adipose tissue; Weighted gene co-expression network analysis
Constructing coexpression networks and performing network analysis using large-scale gene expression data sets is an effective way to uncover new biological knowledge; however, the methods used for gene association in constructing these coexpression networks have not been thoroughly evaluated. Since different methods lead to structurally different coexpression networks and provide different information, selecting the optimal gene association method is critical.
Methods and Results
In this study, we compared eight gene association methods – Spearman rank correlation, Weighted Rank Correlation, Kendall, Hoeffding's D measure, Theil-Sen, Rank Theil-Sen, Distance Covariance, and Pearson – and focused on their true knowledge discovery rates in associating pathway genes and construction coordination networks of regulatory genes. We also examined the behaviors of different methods to microarray data with different properties, and whether the biological processes affect the efficiency of different methods.
We found that the Spearman, Hoeffding and Kendall methods are effective in identifying coexpressed pathway genes, whereas the Theil-sen, Rank Theil-Sen, Spearman, and Weighted Rank methods perform well in identifying coordinated transcription factors that control the same biological processes and traits. Surprisingly, the widely used Pearson method is generally less efficient, and so is the Distance Covariance method that can find gene pairs of multiple relationships. Some analyses we did clearly show Pearson and Distance Covariance methods have distinct behaviors as compared to all other six methods. The efficiencies of different methods vary with the data properties to some degree and are largely contingent upon the biological processes, which necessitates the pre-analysis to identify the best performing method for gene association and coexpression network construction.
The predominant model for regulation of gene expression through DNA methylation is an inverse association in which increased methylation results in decreased gene expression levels. However, recent studies suggest that the relationship between genetic variation, DNA methylation and expression is more complex.
Systems genetic approaches for examining relationships between gene expression and methylation array data were used to find both negative and positive associations between these levels. A weighted correlation network analysis revealed that i) both transcriptome and methylome are organized in modules, ii) co-expression modules are generally not preserved in the methylation data and vice-versa, and iii) highly significant correlations exist between co-expression and co-methylation modules, suggesting the existence of factors that affect expression and methylation of different modules (i.e., trans effects at the level of modules). We observed that methylation probes associated with expression in cis were more likely to be located outside CpG islands, whereas specificity for CpG island shores was present when methylation, associated with expression, was under local genetic control. A structural equation model based analysis found strong support in particular for a traditional causal model in which gene expression is regulated by genetic variation via DNA methylation instead of gene expression affecting DNA methylation levels.
Our results provide new insights into the complex mechanisms between genetic markers, epigenetic mechanisms and gene expression. We find strong support for the classical model of genetic variants regulating methylation, which in turn regulates gene expression. Moreover we show that, although the methylation and expression modules differ, they are highly correlated.
DNA methylation; Gene expression; Association; Epigenetics; WGCNA
Both avian and mammalian basal ganglia are involved in voluntary motor control. In birds, such movements include hopping, perching and flying. Two organizational features that distinguish the songbird basal ganglia are that striatal and pallidal neurons are intermingled, and that neurons dedicated to vocal-motor function are clustered together in a dense cell group known as area X that sits within the surrounding striato-pallidum. This specification allowed us to perform molecular profiling of two striato-pallidal subregions, comparing transcriptional patterns in tissue dedicated to vocal-motor function (area X) to those in tissue that contains similar cell types but supports non-vocal behaviors: the striato-pallidum ventral to area X (VSP), our focus here. Since any behavior is likely underpinned by the coordinated actions of many molecules, we constructed gene co-expression networks from microarray data to study large-scale transcriptional patterns in both subregions. Our goal was to investigate any relationship between VSP network structure and singing and identify gene co-expression groups, or modules, found in the VSP but not area X. We observed mild, but surprising, relationships between VSP modules and song spectral features, and found a group of four VSP modules that were highly specific to the region. These modules were unrelated to singing, but were composed of genes involved in many of the same biological processes as those we previously observed in area X-specific singing-related modules. The VSP-specific modules were also enriched for processes disrupted in Parkinson's and Huntington's Diseases. Our results suggest that the activation/inhibition of a single pathway is not sufficient to functionally specify area X versus the VSP and support the notion that molecular processes are not in and of themselves specialized for behavior. Instead, unique interactions between molecular pathways create functional specificity in particular brain regions during distinct behavioral states.
Understanding how gene transcription relates to behavior is challenging. Learned vocal-motor behavior is a complex trait that represents the output of multiple converging genes, pathways, and patterns of neural activity. Here, we applied a systems analytical approach to determine how thousands of genes change their expression levels simultaneously in a region of the vertebrate brain important for vocal-motor function, the basal ganglia, during a specific vocal-motor behavior, singing. We used the zebra finch species of songbird based on similarities between song learning/production and speech, and because they possess a set of brain subregions dedicated to singing. Microarrays were used to measure gene expression levels in one such song-dedicated region and in an adjacent motor area that is not thought to play a role in vocal function. This allowed us to address the question of whether distinct gene co-expression patterns could be found in each area. We found that each area contained unique patterns of transcriptional co-activity, but there were also unexpected overlaps. We conclude that the particular behaviors (singing versus non-vocal behaviors) supported by these subregions depend on the particular sets of interactions between molecular pathways that occur in each subregion.
Estrogen signaling pathways may play a significant role in the pathogenesis of non-small cell lung cancers (NSCLC) as evidenced by the expression of aromatase and estrogen receptors (ERα and ERβ) in many of these tumors. Here we examine whether ERα and ERβ levels in conjunction with aromatase define patient groups with respect to survival outcomes and possible treatment regimens. Immunohistochemistry was performed on a high-density tissue microarray with resulting data and clinical information available for 377 patients. Patients were subdivided by gender, age and tumor histology, and survival data was determined using the Cox proportional hazards model and Kaplan-Meier curves. Neither ERα nor ERβ alone were predictors of survival in NSCLC. However, when coupled with aromatase expression, higher ERβ levels predicted worse survival in patients whose tumors expressed higher levels of aromatase. Although this finding was present in patients of both genders, it was especially pronounced in women ≥ 65 years old, where higher expression of both ERβ and aromatase indicated a markedly worse survival rate than that determined by aromatase alone. Conclusion: Expression of ERβ together with aromatase has predictive value for survival in different gender and age subgroups of NSCLC patients. This predictive value is stronger than each individual marker alone. Our results suggest treatment with aromatase inhibitors alone or combined with estrogen receptor modulators may be of benefit in some subpopulations of these patients.
NSCLC; tissue microarray; aromatase; estrogen receptor; immunohistochemistry; prognosis
It has been debated whether human induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) express distinctive transcriptomes. By using the method of weighted gene co-expression network analysis, we showed here that iPSCs exhibit altered functional modules compared with ESCs. Notably, iPSCs and ESCs differentially express 17 modules that primarily function in transcription, metabolism, development, and immune response. These module activations (up- and downregulation) are highly conserved in a variety of iPSCs, and genes in each module are coherently co-expressed. Furthermore, the activation levels of these modular genes can be used as quantitative variables to discriminate iPSCs and ESCs with high accuracy (96%). Thus, differential activations of these functional modules are the conserved features distinguishing iPSCs from ESCs. Strikingly, the overall activation level of these modules is inversely correlated with the DNA methylation level, suggesting that DNA methylation may be one mechanism regulating the module differences. Overall, we conclude that human iPSCs and ESCs exhibit distinct gene expression networks, which are likely associated with different epigenetic reprogramming events during the derivation of iPSCs and ESCs.
Many high-throughput biological data analyses require the calculation of large correlation matrices and/or clustering of a large number of objects. The standard R function for calculating Pearson correlation can handle calculations without missing values efficiently, but is inefficient when applied to data sets with a relatively small number of missing data. We present an implementation of Pearson correlation calculation that can lead to substantial speedup on data with relatively small number of missing entries. Further, we parallelize all calculations and thus achieve further speedup on systems where parallel processing is available. A robust correlation measure, the biweight midcorrelation, is implemented in a similar manner and provides comparable speed. The functions cor and bicor for fast Pearson and biweight midcorrelation, respectively, are part of the updated, freely available R package WGCNA.
The hierarchical clustering algorithm implemented in R function hclust is an order n3 (n is the number of clustered objects) version of a publicly available clustering algorithm (Murtagh 2012). We present the package flashClust that implements the original algorithm which in practice achieves order approximately n2, leading to substantial time savings when clustering large data sets.
Pearson correlation; robust correlation; hierarchical clustering; R
Complex traits and other polygenic processes require coordinated gene expression. Co-expression networks model mRNA co-expression: the product of gene regulatory networks. To identify regulatory mechanisms underlying coordinated gene expression in a tissue-enriched context, ten Arabidopsis thaliana co-expression networks were constructed after manually sorting 4,566 RNA profiling datasets into aerial, flower, leaf, root, rosette, seedling, seed, shoot, whole plant, and global (all samples combined) groups. Collectively, the ten networks contained 30% of the measurable genes of Arabidopsis and were circumscribed into 5,491 modules. Modules were scrutinized for cis regulatory mechanisms putatively encoded in conserved non-coding sequences (CNSs) previously identified as remnants of a whole genome duplication event. We determined the non-random association of 1,361 unique CNSs to 1,904 co-expression network gene modules. Furthermore, the CNS elements were placed in the context of known gene regulatory networks (GRNs) by connecting 250 CNS motifs with known GRN cis elements. Our results provide support for a regulatory role of some CNS elements and suggest the functional consequences of CNS activation of co-expression in specific gene sets dispersed throughout the genome.
Lung cancer is the most common cause of cancer mortality in male and female patients in the US. Although it is clear that tobacco smoking is a major cause of lung cancer, about half of all women with lung cancer worldwide are never-smokers. Despite a declining smoking population, the incidence of non-small cell lung cancer (NSCLC), the predominant form of lung cancer, has reached epidemic proportions particularly in women. Emerging data suggest that factors other than tobacco, namely endogenous and exogenous female sex hormones, have a role in stimulating NSCLC progression. Aromatase, a key enzyme for estrogen biosynthesis, is expressed in NSCLC. Clinical data show that women with high levels of tumor aromatase (and high intratumoral estrogen) have worse survival than those with low aromatase. The present and previous studies also reveal significant expression and activity of estrogen receptors (ERα, ERβ) in both extranuclear and nuclear sites in most NSCLC. We now report further on the expression of progesterone receptor (PR) transcripts and protein in NSCLC. PR transcripts were significantly lower in cancerous as compared to non-malignant tissue. Using immunohistochemistry, expression of PR was observed in the nucleus and/or extranuclear compartments in the majority of human tumor specimens examined. Combinations of estrogen and progestins administered in vitro cooperate in promoting tumor secretion of vascular endothelial growth factor and, consequently, support tumor-associated angiogenesis. Further, dual treatment with estradiol and progestin increased the numbers of putative tumor stem/progenitor cells. Thus, ER- and/or PR-targeted therapies may offer new approaches to manage NSCLC.
Progesterone; Estrogen; Steroid hormone receptor; Non-small cell lung cancer; VEGF; Progenitor cells; Cancer stem cells; Angiogenesis
Sjögren’s syndrome is a tissue-specific autoimmune disease that affects exocrine tissues, especially salivary glands and lacrimal glands. Despite a large body of evidence gathered over the past 60 years, significant gaps still exist in our understanding of Sjögren’s syndrome. The goal of this study was to develop a database that collects and organizes gene and protein expression data from the existing literature for comparative analysis with future gene expression and proteomic studies of Sjögren’s syndrome.
To catalog the existing knowledge in the field, we used text mining to generate the Sjögren’s Syndrome Knowledge Base (SSKB) of published gene/protein data, which were extracted from PubMed using text mining of over 7,700 abstracts and listing approximately 500 potential genes/proteins. The raw data were manually evaluated to remove duplicates and false-positives and assign gene names. The data base was manually curated to 477 entries, including 377 potential functional genes, which were used for enrichment and pathway analysis using gene ontology and KEGG pathway analysis.
The Sjögren’s syndrome knowledge base (
http://sskb.umn.edu) can form the foundation for an informed search of existing knowledge in the field as new potential therapeutic targets are identified by conventional or high throughput experimental techniques.
The hallmarks of many haematological malignancies and solid tumours are chromosomal translocations, which may lead to gene fusions. Recently, next-generation sequencing techniques at the transcriptome level (RNA-Seq) have been used to verify known and discover novel transcribed gene fusions. We present FusionFinder, a Perl-based software designed to automate the discovery of candidate gene fusion partners from single-end (SE) or paired-end (PE) RNA-Seq read data. FusionFinder was applied to data from a previously published analysis of the K562 chronic myeloid leukaemia (CML) cell line. Using FusionFinder we successfully replicated the findings of this study and detected additional previously unreported fusion genes in their dataset, which were confirmed experimentally. These included two isoforms of a fusion involving the genes BRK1 and VHL, whose co-deletion has previously been associated with the prevalence and severity of renal-cell carcinoma. FusionFinder is made freely available for non-commercial use and can be downloaded from the project website (http://bioinformatics.childhealthresearch.org.au/software/fusionfinder/).
Despite large-scale genome-wide association studies (GWAS), the underlying genes for schizophrenia are largely unknown. Additional approaches are therefore required to identify the genetic background of this disorder. Here we report findings from a large gene expression study in peripheral blood of schizophrenia patients and controls. We applied a systems biology approach to genome-wide expression data from whole blood of 92 medicated and 29 antipsychotic-free schizophrenia patients and 118 healthy controls. We show that gene expression profiling in whole blood can identify twelve large gene co-expression modules associated with schizophrenia. Several of these disease related modules are likely to reflect expression changes due to antipsychotic medication. However, two of the disease modules could be replicated in an independent second data set involving antipsychotic-free patients and controls. One of these robustly defined disease modules is significantly enriched with brain-expressed genes and with genetic variants that were implicated in a GWAS study, which could imply a causal role in schizophrenia etiology. The most highly connected intramodular hub gene in this module (ABCF1), is located in, and regulated by the major histocompatibility (MHC) complex, which is intriguing in light of the fact that common allelic variants from the MHC region have been implicated in schizophrenia. This suggests that the MHC increases schizophrenia susceptibility via altered gene expression of regulatory genes in this network.
Genomic datasets generated by new technologies are increasingly prevalent in disparate areas of biological research. While many studies have sought to characterize relationships among genomic features, commensurate efforts to characterize relationships among biological samples have been less common. Consequently, the full extent of sample variation in genomic studies is often under-appreciated, complicating downstream analytical tasks such as gene co-expression network analysis.
Here we demonstrate the use of network methods for characterizing sample relationships in microarray data generated from human brain tissue. We describe an approach for identifying outlying samples that does not depend on the choice or use of clustering algorithms. We introduce a battery of measures for quantifying the consistency and integrity of sample relationships, which can be compared across disparate studies, technology platforms, and biological systems. Among these measures, we provide evidence that the correlation between the connectivity and the clustering coefficient (two important network concepts) is a sensitive indicator of homogeneity among biological samples. We also show that this measure, which we refer to as cor(K,C), can distinguish biologically meaningful relationships among subgroups of samples. Specifically, we find that cor(K,C) reveals the profound effect of Huntington’s disease on samples from the caudate nucleus relative to other brain regions. Furthermore, we find that this effect is concentrated in specific modules of genes that are naturally co-expressed in human caudate nucleus, highlighting a new strategy for exploring the effects of disease on sets of genes.
These results underscore the importance of systematically exploring sample relationships in large genomic datasets before seeking to analyze genomic feature activity. We introduce a standardized platform for this purpose using freely available R software that has been designed to enable iterative and interactive exploration of sample networks.
Sample networks; Sample network analysis; Huntington’s disease; Clustering coefficient; cor(K,C); Standardized C(k) curve; Data pre-processing; Microarrays; Gene expression
A critical step in detecting variants from next-generation sequencing data is post hoc filtering of putative variants called or predicted by computational tools. Here, we highlight four critical parameters that could enhance the accuracy of called single nucleotide variants and insertions/deletions: quality and deepness, refinement and improvement of initial mapping, allele/strand balance, and examination of spurious genes. Use of these sequence features appropriately in variant filtering could greatly improve validation rates, thereby saving time and costs in next-generation sequencing projects.
Pathway analysis of a set of genes represents an important area in large-scale omic data analysis. However, the application of traditional pathway enrichment methods to next-generation sequencing (NGS) data is prone to several potential biases, including genomic/genetic factors (e.g., the particular disease and gene length) and environmental factors (e.g., personal life-style and frequency and dosage of exposure to mutagens). Therefore, novel methods are urgently needed for these new data types, especially for individual-specific genome data.
In this study, we proposed a novel method for the pathway analysis of NGS mutation data by explicitly taking into account the gene-wise mutation rate. We estimated the gene-wise mutation rate based on the individual-specific background mutation rate along with the gene length. Taking the mutation rate as a weight for each gene, our weighted resampling strategy builds the null distribution for each pathway while matching the gene length patterns. The empirical P value obtained then provides an adjusted statistical evaluation.
We demonstrated our weighted resampling method to a lung adenocarcinomas dataset and a glioblastoma dataset, and compared it to other widely applied methods. By explicitly adjusting gene-length, the weighted resampling method performs as well as the standard methods for significant pathways with strong evidence. Importantly, our method could effectively reject many marginally significant pathways detected by standard methods, including several long-gene-based, cancer-unrelated pathways. We further demonstrated that by reducing such biases, pathway crosstalk for each individual and pathway co-mutation map across multiple individuals can be objectively explored and evaluated. This method performs pathway analysis in a sample-centered fashion, and provides an alternative way for accurate analysis of cancer-personalized genomes. It can be extended to other types of genomic data (genotyping and methylation) that have similar bias problems.
A novel chemocentric approach to identifying cancer-relevant targets is introduced. Starting with a large chemical collection, the strategy uses the list of small molecule hits arising from a differential cytotoxicity screening on tumor HCT116 and normal MRC-5 cell lines to identify proteins associated with cancer emerging from a differential virtual target profiling of the most selective compounds detected in both cell lines. It is shown that this smart combination of differential in vitro and in silico screenings (DIVISS) is capable of detecting a list of proteins that are already well accepted cancer drug targets, while complementing it with additional proteins that, targeted selectively or in combination with others, could lead to synergistic benefits for cancer therapeutics. The complete list of 115 proteins identified as being hit uniquely by compounds showing selective antiproliferative effects for tumor cell lines is provided.
Amyotrophic lateral sclerosis (ALS) is a progressive, neurodegenerative disease characterized by loss of upper and lower motor neurons. ALS is considered to be a complex trait and genome-wide association studies (GWAS) have implicated a few susceptibility loci. However, many more causal loci remain to be discovered. Since it has been shown that genetic variants associated with complex traits are more likely to be eQTLs than frequency-matched variants from GWAS platforms, we conducted a two-stage genome-wide screening for eQTLs associated with ALS. In addition, we applied an eQTL analysis to finemap association loci. Expression profiles using peripheral blood of 323 sporadic ALS patients and 413 controls were mapped to genome-wide genotyping data. Subsequently, data from a two-stage GWAS (3,568 patients and 10,163 controls) were used to prioritize eQTLs identified in the first stage (162 ALS, 207 controls). These prioritized eQTLs were carried forward to the second sample with both gene-expression and genotyping data (161 ALS, 206 controls). Replicated eQTL SNPs were then tested for association in the second-stage GWAS data to find SNPs associated with disease, that survived correction for multiple testing. We thus identified twelve cis eQTLs with nominally significant associations in the second-stage GWAS data. Eight SNP-transcript pairs of highest significance (lowest p = 1.27×10−51) withstood multiple-testing correction in the second stage and modulated CYP27A1 gene expression. Additionally, we show that C9orf72 appears to be the only gene in the 9p21.2 locus that is regulated in cis, showing the potential of this approach in identifying causative genes in association loci in ALS. This study has identified candidate genes for sporadic ALS, most notably CYP27A1. Mutations in CYP27A1 are causal to cerebrotendinous xanthomatosis which can present as a clinical mimic of ALS with progressive upper motor neuron loss, making it a plausible susceptibility gene for ALS.
Major depression is nearly twice as prevalent in women compared to men. In bipolar disorder, depressive episodes have been reported to be more common amongst female patients. Furthermore, periods of depression often correlate with periods of hormonal fluctuations. A link between hormone signaling and these mood disorders has, therefore, been suggested to exist in many studies. Estrogen, one of the primary female sex hormones, mediates its effect mostly by binding to estrogen receptors (ERs). Nuclear ERs function as transcription factors and regulate gene transcription by binding to specific DNA sequences. A nucleotide change in the binding sequence might alter the binding efficiency, which could affect transcription levels of nearby genes. In order to investigate if variation in ER DNA-binding sequences may be involved in mood disorders, we conducted a genome-wide study of ER DNA-binding in patients diagnosed with major depression or bipolar disorder. Association studies were performed within each gender separately and the results were corrected for multiple testing by the Bonferroni method. In the female bipolar disorder material a significant association result was found for rs6023059 (corrected p-value = 0.023; odds ratio (OR) 0.681, 95% confidence interval (CI) 0.570–0.814), a single nucleotide polymorphism (SNP) placed downstream of the gene coding for transglutaminase 2 (TGM2). Thus, females with a specific genotype at this SNP may be more vulnerable to fluctuating estrogen levels, which may then act as a triggering factor for bipolar disorder.
Symptoms of Major Depressive Disorder (MDD) are hypothesized to arise from dysfunction in brain networks linking the limbic system and cortical regions. Alterations in brain functional cortical connectivity in resting-state networks have been detected with functional imaging techniques, but neurophysiologic connectivity measures have not been systematically examined. We used weighted network analysis to examine resting state functional connectivity as measured by quantitative electroencephalographic (qEEG) coherence in 121 unmedicated subjects with MDD and 37 healthy controls. Subjects with MDD had significantly higher overall coherence as compared to controls in the delta (0.5–4 Hz), theta (4–8 Hz), alpha (8–12 Hz), and beta (12–20 Hz) frequency bands. The frontopolar region contained the greatest number of “hub nodes” (surface recording locations) with high connectivity. MDD subjects expressed higher theta and alpha coherence primarily in longer distance connections between frontopolar and temporal or parietooccipital regions, and higher beta coherence primarily in connections within and between electrodes overlying the dorsolateral prefrontal cortical (DLPFC) or temporal regions. Nearest centroid analysis indicated that MDD subjects were best characterized by six alpha band connections primarily involving the prefrontal region. The present findings indicate a loss of selectivity in resting functional connectivity in MDD. The overall greater coherence observed in depressed subjects establishes a new context for the interpretation of previous studies showing differences in frontal alpha power and synchrony between subjects with MDD and normal controls. These results can inform the development of qEEG state and trait biomarkers for MDD.
Colorectal cancer (CRC) is one of the leading malignant cancers with a rapid increase in incidence and mortality. The recurrences of CRC after curative resection are sometimes unavoidable and often take place within the first year after surgery. MicroRNAs may serve as biomarkers to predict early recurrence of CRC, but identifying them from over 1,400 known human microRNAs is challenging and costly. An alternative approach is to analyze existing expression data of messenger RNAs (mRNAs) because generally speaking the expression levels of microRNAs and their target mRNAs are inversely correlated. In this study, we extracted six mRNA expression data of CRC in four studies (GSE12032, GSE17538, GSE4526 and GSE17181) from the gene expression omnibus (GEO). We inferred microRNA expression profiles and performed computational analysis to identify microRNAs associated with CRC recurrence using the IMRE method based on the MicroCosm database that includes 568,071 microRNA-target connections between 711 microRNAs and 20,884 gene targets. Two microRNAs, miR-29a and miR-29c, were disclosed and further meta-analysis of the six mRNA expression datasets showed that these two microRNAs were highly significant based on the Fisher p-value combination (p = 9.14×10−9 for miR-29a and p = 1.14×10−6 for miR-29c). Furthermore, these two microRNAs were experimentally tested in 78 human CRC samples to validate their effect on early recurrence. Our empirical results showed that the two microRNAs were significantly down-regulated (p = 0.007 for miR-29a and p = 0.007 for miR-29c) in the early-recurrence patients. This study shows the feasibility of using mRNA profiles to indicate microRNAs. We also shows miR-29a/c could be potential biomarkers for CRC early recurrence.
Induced pluripotent stem (iPS) cells can be obtained from fibroblasts upon expression of Oct4, Sox2, Klf4 and c-Myc. To understand how these factors induce pluripotency, we carried out genome-wide analyses of their promoter binding and expression in iPS and partially reprogrammed cells. We find that target genes of the four factors strongly overlap in iPS and embryonic stem (ES) cells. In partially reprogrammed cells, many genes co-occupied by c-Myc and any of the other three factors already show an ES-like binding and expression pattern. In contrast, genes that are specifically co-bound by Oct4, Sox2 and Klf4 in ES cells and encode pluripotency regulators severely lack binding and transcriptional activation. Among the four factors, c-Myc promotes the most ES cell-like transcription pattern when expressed individually in fibroblasts. These data uncover temporal and separable contributions of the four factors during the reprogramming process and indicate that ectopic c-Myc predominantly acts before pluripotency regulators are activated.

domain: literature


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Well, I finally decided to take the plunge. For several years now, I have been toying around with this idea, but never could seem to commit to anything. But I finally decided it's time. I am now officially going vegetarian! YAY for me!!
So why in the world would I do this? I have several reasons. But the one most prevalent is my stomach. I guess for about the last 3-4 years I have just had one stomach problem after another. My belly constantly HURT! I did TONS of research, and I realized that meat and dairy products are some of the most problem causing foods. One of the reasons meat can give you so much trouble is the simple fact that it doesn't digest well in your stomach. It just sits there for days before it passes out of you. Which means for me, it was sitting there causing my stomach to hurt for days. And every time I would eat more meat, it just compounded the problem. The thing, for me at least, about dairy is the protein that is found in cows milk. There is just an amazing number of people who have a milk protein allergy, and I happen to be one of them. Every time I would eat a bowl of cereal or chow down on some delicious dutch chocolate ice cream, the protein in the dairy would just make my tummy SCREAM! Once I gave up eating red meat and dairy products, my tummy was feeling much better - but not completely. About 2 months ago I decided to try going without any chicken to see if the complete absence of meat would make a difference, instead of just doing without the red meat. Now, my tummy is even BETTER!! (Oh and just so you know, pork is just as bad, only I don't eat it anyway because it tastes like DIRT to me. Probably because the little porkers live in it all the time!)
So here I am. No meat and no dairy, and I feel worlds better!! In truth, I am working towards a complete vegan diet, but as of right now, I'm not there yet. There are 2 main things keeping me from saying I am a vegan. The first is the fact that I still eat eggs. I get my eggs from my daddy, who happens to treat his hens like royalty. So I know these gals aren't being mistreated just so I can have an egg from them. And the second thing is that I am not giving up honey. I happen to be very good friends with my bee keeper (she and I have been friends since high school), and I know for a fact that she takes really great care of her hives. So I know eating her honey is perfectly great for me. Not to mention the fact that I LOVE the stuff, and it is quite medicinal, so I don't plan to stop with it. Those are 2 things that vegans don't consume, so I guess I'll just have to say I'm sticking with being a vegetarian. Oh and I also occasionally still sprinkle some cheese on my food. Vegan cheese is expensive, so I just don't use it often. But occasionally I just really want some cheese on top of my baked potato or soup!
And what am I supposed to eat?? Oh my goodness, there is a world of great food out there! When you eat mainly a meat centered diet, you seem to limit the "extra" foods you are willing to try. You just pull into the first fast food restaurant you come to and order the same thing - burger and fries - and you think that's all there is. Well let me tell you, there is so much more to life and eating than burgers and fries!
One of my favorite websites is Vegetarian Times. If you spend only a few minutes perusing their site, you will find a plethora of yumminess that you just must try! Even the most die hard meat eater will want to sample some of the dishes, if only as a side dish. Believe me, I didn't start out with eating like this completely. Everything I tried was considered a side dish, because I felt like, if it bombed, at least we had our other food to eat. But now I find myself making more and more great veggie dishes, and they are now my main course. And oh the variety!
A book that I read recently was The Kind Diet by Alicia Silverstone. That is one AMAZING book, and I HIGHLY recommend it to anyone thinking of a change in lifestyle where your diet is concerned, or if you're just curious about why anyone would choose to live a veggie lifestyle. You can also check out her website - The Kind Life - for more recipes, info, and anything else you may be wondering.
Also, vegetarian items are showing up more and more in grocery stores everywhere! I can get vegetarian meat from Morningstar Farms that is just fantastic! They have the greatest veggie burgers that are so full of flavors and variety. Meat pretty much has one flavor, but not these tasty veggie burgers! I buy Rice Dream rice milk and Almond Breeze almond milk to drink, both of which are at my local Wal-Mart. Both of these companies sell some of the greatest dairy free ice cream, too!! YAY for ice cream!
So I say all that to say, I am really excited about my new vegetarian diet! It's still a work in progress. The rest of my family is not going this route with me, and that's OK. I still cook plenty of meat for them. So all of you meat eaters out there, don't worry! You don't need to invite my husband and kids over for supper. I'm still feeding them! But I will say that veggies are becoming the bigger part of our meals, and meat is taking a back seat for everybody. And as long as I keep cooking really tasty foods, no one is complaining!
One more change I am heading toward but haven't reached as of yet is a gluten free diet. Even after cutting so much from my diet, I still find I have stomach aches often. Even though my tummy feels worlds different from when I ate meat and dairy, it still isn't enough. I have too much to do in this life to be held back because of feeling lousy. So I am leaning towards going gluten free. But for right now, I'm sticking with the newest changes I have made and just getting used to them for awhile. It's a process. Good health and changing a lifestyle of bad habits doesn't happen overnight. It takes time. This change has taken a long time. But here I am now, and I am feeling all the better for it!
Have you made any lifestyle changes where your eating is concerned? Have a great vegetarian or gluten free recipe you want to share to help me out here? Leave me a comment and tell me all about it!


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Merger of two recycling associations would lead to the formation of the NorthAmerican Recycling Coalition.
The National Recycling Coalition (NRC) and the Recycling Organization of North America Inc. (RONA) have announced a significant step toward consolidating the two associations. If the merger is complete, the result would be the creation of a new organization called the North American Recycling Coalition.
According to the NRC, both associations are already providing services, representation, institutional knowledge and staff to their individual and shared members. The intent is to unify the efforts in one body. Through consolidation, both organizations would build on each other's successes, resources, assets, vision, and goodwill to build a new coalition, the NRC speculates.
To accomplish this, boards of both associations have been meeting to finalize the creation of the new NRC. While acknowledging significant work that needs to be addressed, the NRC is calling a special meeting of the association July 13. Additionally, the association is taking actions to amend its current New York State Certificate of Incorporation to change the National Recycling Coalition name to that of North American Recycling Coalition (NRC).
The associations say they expect to make a formal announcement on the merger at the Resource Recycling Conference in Austin, Texas, this August.


Recently I was invited to attend a concert at a venue new to me. It was a large space, well lit, with great acoustics, very warm and inviting–like sitting in an incredibly large living room, including plush leather chairs. I noticed there were a lot of microphones set up, and assumed they were making a recording of the evening–which is fine.
However, it seemed that the audience was not really included in the performance. As a classical music performer, I know that it isn’t customary to speak to the audience during the concert. In this case, they started with a selection of art songs sung in German, with neither a verbal explanation of the text nor printed concert notes. The audience never knew what the songs were about, nor when the songs ended–apart from the heavy sigh from the singer as she turned to the next page. At the end of the fifteen minute set, the vocalist–who sang beautifully, I might add–closed all her songbooks, and gave the audience a quick nod, to let us know she was finished.
Next the pianist played a selection of short pieces by a contemporary composer, who then rose from the audience himself, to do 3 more of his compositions. To his credit, the composer did give us the names of each piece as he played them, but again, the audience sat there wondering what was going on, and hoping we would be able to guess when the pieces were over. Overall, it was a puzzling and frustrating experience.
My first voice teacher told me that when you bow at a concert you are bending your head, as if you were allowing the audience to pat you on the head. In this case, the audience felt like the performers flinched out of the way before we could touch them.
I learned in discussing the concert afterwards with one of the performers and the sound engineer that they had been trying to save editing time post concert, by having minimal clapping and speech between numbers as they made the recording. OK, I understand what they were trying to do. But I felt cheated. The audience had expended time and effort and money to be there to support them, but they were shut out of the performance. I don’t understand why they chose to have an audience at all. They were making a studio recording, with the audience as decoration–but they didn’t take photos either, so the audience had no contribution to the process at all. It was a great shame, because the performers really did a great job, and it would have been nice to be able to let them know it.
I once attended a concert at Carnegie Hall, where the guest artist announced sternly at the beginning of the concert that they were recording the evening for an album, and wanted the audience to keep absolutely still, and not cough or turn pages of the program during the pieces, because it would ruin the recording. We all sat there in terror during the first piece, then the audience EXPLODED into fits of coughing when the conductor signaled they had finished. Some people rushed for the exits to escape….At intermission there was a mass exodus…Again, if they didn’t want an audience, why book a venue rather than a studio? I suppose, the income from ticket sales is the reason, but I wouldn’t buy the album afterwards…
Just a thought….


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The 22-year-old Abreu, the No. 5-ranked prospect in the organization according to Baseball America, was having one of the most impressive camps on the club. He had just singled in the bottom of the seventh inning, raising his Spring Training batting average to .394. His 10 RBIs are tied for the club lead. He played last year at Double-A Jacksonville.
Bet on it: Wilson Betemit had the game-winning hit in Saturday's 7-5 victory over Houston, and he did it right-handed, bouncing a tie-breaking double down the third-base line off former Dodgers reliever Kelly Wunsch in a three-run eighth inning.
"I know I can hit right-handed," said the switch-hitting Betemit, who could do it at only a .189 clip last year. "It's coming. I have to be patient. Right-handed, I lean and jump at the ball. Eddie [Murray, hitting coach] tells me to stay back, but hit the ball in front of the plate, don't wait for it to come too deep.
"I also know that I move my head too much. I watch Olmedo Saenz hit and he's very quiet at the plate and that's how I want to be."
After a very slow start, Betemit's spring average is up to .250 with two hits and a walk Saturday.
"Lately, I like just about everything he's doing, to tell you the truth," said manager Grady Little. "He's been playing good baseball the last week. He's turned it up a bit. He's getting hits, right-handed as well as left-handed, and making plays. That's all we ask."
Little said if the season were to open Sunday, Andre Ethier would bat seventh in the order behind Russell Martin and Betemit would bat eighth.
"I like the way the lineup looks, regardless of the bottom," he said.
Earlier in the game, the top two of Little's order went 4-for-6, with Rafael Furcal and Juan Pierre getting two hits each, Pierre driving in a pair with a run and Furcal scoring twice.
Brett Tomko burned up his pitch count in three innings, limiting the damage to only one run.
"He had a rough start, but he settled in and pitched okay," said Little.
The best outing of the day belonged to forgotten man D.J. Houlton, with two perfect innings that dropped his ERA to 1.35. Joe Mays was charged with three runs, looking better at the end than the beginning. Matt White allowed a home run in the ninth but got the save.
Simulation time: Reliever Joe Beimel was on the back field Saturday in a simulated game against Minor Leaguers, throwing 35 pitches in two innings, on breaking balls particularly.
"It was very productive," said Beimel. "I don't like giving up runs in games or walking guys and in this setting, you can fall behind in the count and throw a breaking ball and not worry about it. I like these simulated games. When you get in even an exhibition game, the competitiveness kicks in and don't want to get ripped around so you don't work on things that need working on."
Beimel said he has no lingering effects of the hand gash that wiped out his postseason.
"It's a non-issue," he said. "I feel pretty much ready for the season. The last game I pitched in, everything started to click. My arm speed was better and the pitches were crisper."
Injury update: Jason Repko ramped up his rehab from a strained groin muscle, sprinting straight ahead, working on turns and taking fly balls.
Duncan excused: First-base coach Mariano Duncan was excused for the weekend because of personal business. Mark Weidemaier, special assistant to the general manager, coached first base in the game.
World Congress of Sports: Dodgers Vice Chairman and President Jamie McCourt will take part in a panel discussion on sports, entertainment and some of the "hot-button" issues in today's news on Wednesday, March 21, presented by Street & Smith's Sports Business Journal/Daily. The event, which will take place in New York, features McCourt, NCAA president Myles Brand, executive vice president of Business for Major League Baseball Tim Brosnan, St. Louis Blues owner Dave Checketts and executive vice president of content for ESPN John Skipper.
Coming up: Opening Day starter Derek Lowe and Chad Billingsley are scheduled to pitch Sunday in Winter Haven against Cleveland, but the rest of the travel squad is light on regulars, with catcher Russell Martin and right fielder Andre Ethier the only ones scheduled to play.


Most of Scotland could be hit by ice tomorrow, affecting driving conditions, the Met Office has warned.
The Yellow warning of ice is in place for tomorrow morning, running from 1am on Saturday until 11am, with the official forecaster warning of an increased risk of injuries or accidents.
The warning, which comes after the country was hit with snow and rain earlier in the week, is in place for Strathclyde, the Highlands, Lothiian and Borders, and Central, Tayside and Fife.
Much of Northern England is also expected to be hit by the ice, which is likely to affect travelling conditions.
The Chief Forecaster’s assessment read: “As we enter a spell of cold weather, a band of rain and snow will move into the UK overnight and into Saturday. Over high ground in Scotland and across the higher Pennine routes, snowfall may amount to 2 or 3 cm in places.
However the main hazards seem more likely to come from ice, where rain falls on frozen surfaces.”


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Dikkie Dik Flowers and butterflies window drawing
During Spring, Dikkie Dik loves to sit in the flowers and watch the butterflies, you too? Sweet window drawing, illustrated by Jet Boeke.
Love this style? Go over here to find all Dikkie Dik window drawing designs in the shop.
design ©Jet Boeke / Dikkie Dik
photo Esther W, Shandy B and Manon K
file(s) 72 dpi PDF / mirrored n/a
download on computer/laptop, tablet/phone not possible
sizes ± (all included)
S - 15 x 18,5 cm
M - 20 x 25 cm
L - 27,5 x 34,5 cm
XL - 36 x 45 cm
XXL - 48 x 60 cm
This is not a physical product, but a digital design you print (at home). Direct download after order is completed (up to 3 downloads, but print as often as you like). All designs are only for personal use.


Thousands Of Imported Models In Tyre Database
Thousands Of Imported Models Listed In Unique Tyre Database
THOUSANDS OF IMPORTED MODELS LISTED IN UNIQUE TYRE DATABASE
Need a tyre for a 1994 McLaren, or a 1999 Autozam Laputax? Any idea what size?
A call to your nearest Beaurepaires, Goodyear Auto Service Centre or Frank Allen Tyres Store should yield the correct details.
They have just installed New Zealand Tyre Match, a powerful new database developed over the past twelve months by South Pacific Tyres NZ Ltd.
The huge range of used imported Japanese cars had made specifying the correct tyres a minefield, product development manager Bill Prebble said.
"Fitting the correct tyre is very important to the handling and safety of any car or recreational vehicle.
"Matching tyres is critical on 4WD recreational vehicles as different diameter tyres either side to side or front and back can create mechanical problems when the vehicle is operated in four wheel drive.
"All our retail stores are on line to a database that covers the last six years of Japanese vehicle production and includes the original equipment fitment, the overall diameter of the OE tyres when new and specifies tyres of similar overall diameter suitable for replacement.
"We now have over 8000 different Japanese models listed. In addition we have data for all vehicles manufactured in New Zealand over the past 15 years and list data for product imported from other countries too.
"The New Zealand Tyre Match database is now live on the computer at every one of our retail outlets. It's a tremendous tool for our forecourt staff and motorists can feel re-assured that the correct tyres are being fitted to their vehicle."
FROM SOUTH PACIFIC TYRES NZ


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A father and girlfriend have been arrested on charges for a deadly canal crash that killed two young boys.
Following an investigation, Cape Coral police arrested the father, Kenneth Wayne Lawson, on two counts of DUI manslaughter after a car crashed into a Cape Coral canal and killed his two sons, 10-year-old John Wayne and 7-year-old Titus.
CCPD also announced Friday afternoon that girlfriend, Julia Ann Drudy, who was also in the car at the time of the crash was wanted. She was arrested later that night.
Lawson faces separate charges in Lee County related to property damage at a woman’s home. He is also wanted on charges in Michigan. CCPD confirmed Lawson will be prosecuted in Florida before he would see extradition to Michigan for charges there. However, Florida would have to pay to extradite him, since investigators there won’t.
On Saturday, both had a first appearance in court. Lawson is held with bail set at $525,000; $250,000 for each DUI manslaughter count and another $25,000 for a suspended license charge. Drudy has bail set at $10,000. Both are restricted by the court from the possession of, or consuming, alcohol or drugs.
“The prosecutor and the police have made the decision that, if he is caught out of state, they are not going to extradite him,” Swartz explained to WINK News previously. “Police officers have to come down here and transport the defendant up there, and it’s all a very expensive proposition.”
A family member told us they are happy justice will be served for the two boys.
Acting CCPD Chief Anthony Sizemore says the officers and the good Samaritan felt the same way the community did — anger over the deaths of the brothers and a need for justice.
“Especially for our investigators and our team, I mean, we’re mothers and fathers and have children as well, and the firefighters who assisted and the good Samaritan,” Sizemore said. It is just tragedy upon tragedy.”
Cape Coral Police Department arrested Julia Ann Drudy, 31, Friday night.
According to police, Drudy spoke with officers after seeing the press conference and wanted to turn herself in.
Drudy was arrested and faces charges for permitting an unauthorized operator to drive. She is in the process of being taken to Lee County Jail for booking.
With the arrests made, community members continue to pray for the two brothers. Good Samaritan Tyler Ravert jumped into the canal and did everything he could to save John Wayne and Titus from the car they were trapped inside.
“He (Lawson) put me in that situation. He put my wife in that situation,” Ravert said. “This is a very personal thing to me.”
During the conference for Lawson’s arrest, acting Chief Sizemore explained how these investigations can affect the public and the police department.
“I realize it can be frustrating as we wait for an arrest, but we sacrifice a quick arrest for a thorough investigation,” Sizemore said.
Police say Lawson and Drudy got out of the car after the crash and left the boys behind. That’s something Ravert doesn’t accept.
“It’s constantly in my head, just constantly, and it’s kind of like put a shadow over everything,” Ravert said. “This is like all I’m focusing on. It’s just for those children.”
You can watch police announce Kenneth Lawson’s arrest below, or follow this link:

domain: law


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Stay at home restrictions are tough for everyone. Today, I have collected some free e-books for children and adults that are a gift by writers to help you through lockdown, easy your journey and also to help take your mind away from your troubles for a while.
Did you take up the #HealthyAtHome challenge by the WHO? I did. If you haven't heard about it, you can see here what the Healthy At Home challenge is all about. I did this video on 10 ways you didn't think of to prevent CORONAVIRUS infection. We are all being very careful, and that is... Continue Reading →
As my country goes under lockdown, I have started The Lockdown Journal to document this epic battle the world is fighting against this pandemic. Through my writing, I aim no provide a place to share our news and stories, to give support to those who are feeling stressed, to show that none of us is alone in this. We are all here for each other.
Who are you? You are the sunshine and the dark night, a tempest and the gentle spring. You are every colour of the rainbow. Hold your head high, wear your crown with grace and remember, you are enough. You are worth it. You are a woman, and that is your power!
Celebrations!!!! My latest book LOVE SYMPHONY is #1 New Release on Amazon! I am over the moon... It is a happy moment to have my second book after Breathless to touch the top spot. God is good! Thank you, everyone, for your awesome support!
It's the season of love and it's raining freebies!
Just for this weekend, that is 8th and 9th of February, 2020, you can get the "Love Symphony" kindle book absolutely free on Amazon. All you need to do is visit your Amazon account and click on the buy link when you see the kindle price mentioned as zero.
This book is also free for the subscribers of Kindle Unlimited.
Who doesn't love E-cards! And when they are free to use...that's a wish come true.
Now, send your thoughts and wishes to your loved ones with these free video and animated cards.
Hi friends Sometimes life reminds us to take it slow... Until the bug playing in my veins decides to leave, I would not be able to get to your posts much. But I will be back very soon. Until then, keep showing up here, it feels wonderful to have such beautiful people around. Happy days... Continue Reading →


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One of the great examples of literary advice-giving took place in the summer of 1878. Guy de Maupassant was on the verge of becoming famous. As Flaubert’s literary nephew, and a member of the new group calling themselves Naturalists, he was already well known in Paris; three years previously, he had made his first appearance – as ‘le petit Maupassant’ – in the Goncourt Journal, delighting a company of already famous writers with a long story about Swinburne’s decadent behaviour in Etretat. He had written poems, stories and journalism, coauthored a lewd play, and was working on his first novel, Une Vie. He was socially and sexually successful, and physically very fit: the previous summer, having bought a small boat on Zola’s behalf, he had rowed it the 50 kilometres from Bezons to Zola’s house at Médan. Yet on 3 August, two days before his 28th birthday, he made the following complaints to Flaubert about life: ‘Fucking women is as monotonous as listening to male wit. I find that the news in the papers is always the same, that the vices are trivial, and that there aren’t enough different ways to compose a sentence.’
more from Julian Barnes at the LRB here.


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At 10 Weeks Old: Sophie is still doing great! She is sleeping through the night in her crate without crying. She goes outside to go potty 80% of the time and only uses the litter box occasionally now. She is learning to lots of things, and can already do sit, lay down, and come. We are working on come with distractions now! They finished the fence in our yard last week and she LOVES it. She runs all over the backyard. She also loves to sit on our deck and watch all the wildlife (ducks, squirrels, birds). We are so impressed that she doesn’t bark at all! She just takes it all in. We are starting to work on leaving her for short periods of time, which is hard for her since she has been with us every second for the past month. We started yesterday with us all going upstairs for 5 minutes while she was in her downstairs crate. She hated it and cried, but hopefully she will get used to it. Luckily for her she will never have to be left alone for long!
Thanks again for all the great information you guys send! – Kari & Mark
Congrats to Kari and Mark in Ohio on their new doodle girl! Sophie is a female black phantom AussieDoodle puppy with a soft black based coat and vivid copper accents. Her copper should fill in more around her precious little muzzle as her hair gets longer. We think she will look a lot Stormy as she grows and gets fluffy. Such sweet little honey isn’t she?

domain: pet