VIRUS AID: Direct deposits of virus aid money began going into bank accounts on April 12. If you paid your 2018 or 2019 taxes electronically or receive direct deposit Social Security, SSDI, survivor benefits, or Railroad Retirement benefits, the money will go straight into your bank account.
If you filed taxes and got a check in the mail, you’ll get the money the same way and it’ll be a while.
People who don’t normally file taxes can go to the link below for instructions.
MEDIA MALPRACTICE: The Slime Stream Media Talking Point of the Day was “grim new milestone” –
- ABC reported, “Tonight, the grim milestone. As we come on, America now the deadliest nation in confirmed coronavirus cases. More than 20,000 have died from COVID-19, the U.S surpassing Italy.“
- CBS reported, “Tonight, a grim new milestone — the U.S. now leads the world in coronavirus deaths, more than 20,000 and counting. Worldwide coronavirus deaths now top 107,000, and today the United States surpassed Italy as the country with the most deaths. So far, about 20,000 Americans have died.”
- NBC reported, “Breaking tonight, the devastating toll of the coronavirus — 2,000 Americans killed in one day — more people have died now in the U.S. than any other country. … The U.S. reached a new grim milestone today — more than 20,000 Americans dead by the virus, now the highest death toll in the world.”
Missing in these look-alike reports (who does write these memos?!) were two important points.
A. China has been grossly under-reporting their numbers and almost certainly has the highest death toll in the world.
B. On a per capita basis, most of the countries in Western Europe have higher reported death rates from the virus than the we do. For example, as of Sunday, April 12, the per capita death toll stands at:
- USA = 6.61 deaths per 100,000 residents.
- UK = 15.84 deaths per 100,000 residents (more than double USA rate).
- France = 21.48 deaths per 100,000 residents (more than triple USA rate).
Italy, Spain, Switzerland, Belgium, and the Netherlands also all have per capita death rates that are substantially higher than ours is.
KENTUCKY: On Good Friday, Kentucky’s Democratic governor, Andy Beshear, announced that authorities would be recording the license plates of everyone who attends a “mass gathering” (read: Easter service). Local health departments will then come to the homes of every one of those nasty Christians with a mandatory order to self-quarantine for 14 days.
Kentuckians need to remember this in November.
MICHIGAN: Over the weekend, Michigan’s Democratic governor, Gretchen Whitmer, instituted an expansive new set of restrictions requiring big box stores like Walmart and Target to shut down sections dedicated to items that she considers nonessential and forbidding visits between households.
In case you misunderstood, that is a great bit “Screw you!” to Michiganders who want to plant their gardens, work on DIY home improvement projects, check up on relatives, or help out their elderly neighbors during the quarantine. Cuz, Governor Whitmer says you’re too stupid to figure out how to do any of that safely.
Rep. Justin Amash (I-Mich.) tweeted, “I have a constitutional duty to ensure states don’t trample on the rights of the people. @GovWhitmer’s latest order goes too far and will erode confidence in her leadership. She should immediately reassess it.”
Michiganders need to remember this in November.
CANADA: The Royal Canadian Mounted Police announced they will be making home visitations to check compliance with the Quarantine Act, which requires all Canadians who have come from abroad to self-quarantine for two weeks upon arrival. Fines for non-compliance will be $750,000 to $1 million Canadian dollars and three years in jail.
RCMP detectives will use arrest only as a last resort, as the agency wants to protect its officers from infection with the virus as much as possible. Preferably, a summons or notice will be issued with a future court date for the defendant to appear.
“Our officers will speak to the person, inform them of the law, and explain the importance of compliance, as well as the potential consequences of non-compliance,” the RCMP statement further read.
CHINA: [3:50] – Days after lock down measures were relaxed in virus epicenter Wuhan, a province in northern China is preparing thousands of hospital beds in anticipation of a surge of CCP virus patients.
Not everything is about the virus …
OBAMA/BIDEN: [2:39] – Biden has been slamming Trump for the shortage of PPEs, but it was the Obama administration’s use of AND FAILURE TO RESTOCK the strategic national stockpile that left us with critical shortages of masks, gloves, etc.
We all need to remember this in November.
MEDIA MALPRACTICE: [3:22] – On Easter Sunday, the New York Slime published the result of its investigation into Tara Reade’s accusation that Joe Biden had sexually assaulted her in 1993 when she was working in his U.S. Senate office.
The story said, “We found no pattern of sexual misconduct by Biden, beyond hugs, kisses and touching that women previously said made them uncomfortable.”
Readers pointed out that hugging, kissing, and touching women who do not want to be hugged, kissed, or touched is the very definition of sexual misconduct.
The New York Slime edited its story, which now says only, “We found no pattern of sexual misconduct by Mr. Biden.”
TRUMP’S TAXES: And another hoax bites the dust – Democrats tried to force President Trump to turn over six years of his tax returns to Congress.
When Treasury Secretary Steven Mnuchin and other department officials told them to go pound sand, House Ways and Means Chairman Richard Neal (D-Mass.) requested the Treasury Department’s Inspector General (TIG) investigate.
On April 8, the TIG issued its report finding no wrongdoing.
BERNIE BROS: No on Joe – The Democratic Socialists of America (DSA) tweeted, “We are not endorsing [Joe Biden].”
The DSA is an ultra-progressive group with roughly 56,000 members.
VIRGINIA: On Good Friday, Virginia’s Democratic governor, Ralph Northam, signed another bunch of new and even stricter gun control measures into law.
The laws—which go into effect on July 1—include universal background checks on all gun sales, a limit on handgun purchases to one per month, a law penalizing gun owners for not reporting a stolen firearm, and a “red flag” bill that would allow authorities to temporarily confiscate guns from anyone deemed by a judge to be dangerous to themselves or others.
Virginians need to remember this in November.
HATE GROUP: The Southern Poverty Law Center (SPLC) Hatewatch division maintains a watch list of “hate groups” that supposedly promote violence.
The site’s banner is more honest about what it is really all about: “Hatewatch is a blog that monitors and exposes the activities of the American radical right.”
The site is really about targeting and harassing groups THEY hate. Among the “activities of the American radical right” that they oppose are promoting traditional marriage, professing traditional Christianity, and holding to conservative political positions.
E.g., if you oppose open borders and illegal immigration, they call you a “nativist hater,” “anti-immigrant” and/or “white nationalist.”
Their latest target is the American Family Association (AFA) which promotes traditional marriage, asks Americans to pray for President Trump, and maintains a list of companies that do and do not support Christian values.
In an article on the SPLC Hatewatch site, they lump the AFA in with neo-Confederates and Ku Klux Klanners, one of whom was sentenced to 25-months for his role in a gang-beating of a black man.
ANTI-CHRISTIAN HATE: This past year of anti-Christian hate on television has been overwhelming. The faithful have been called “right-wing ding dongs,” “idiots,” “suckers,” “mental,” and “stupid people” for their belief in God. The Bible was called “male revisionist history” that needed an “upgrade” like an iPhone. And, of course, there was a lot of sexualization of religious themes.
Fox Drama gave Catholics an extra big bashing on its Star finale last year. The awards show featured singer Lil Dini, shirtless with a bunch of crucifixes hanging from his neck, performing alongside a giant light-up cross and a likeness of himself cloaked with a halo behind his head and Rapper Rashad, dressed like the pope with a backup choir dressed in white and scantily clad “nuns” gyrating and shaking their behinds at him.
DOGS BREAKFAST: [1:31] – The pandemic has left professional sportscaster Andrew Cotter without any professional sports to commentate … so he’s making do with what’s around home, as so many of us are.


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text:
From The Guardian (18 June 2007)
Greece is mounting a nationwide effort to remove “eye candy” billboards from roadsides, amid growing evidence that images of women wearing not very much contribute to Europe’s worst road accident figures.
With 15,000 hoardings in the capital alone, drivers are distracted by “unacceptable levels of eye candy”, say campaigners who have convinced the courts to rule that all roadside adverts be dismantled.
Billboards invariably depict svelte females in outre poses. “Many of them not only hide traffic lights and road signs, they are put up illegally,” said an Athens traffic police official.
“We believe they are the cause of 10% of all accidents in the city.”
Driving in Greece is not for the faint-hearted. More than 2,000 people die on the roads annually; another 4,000 are seriously injured in 22,000 car accidents a year – one every 24 minutes.
The Foreign and Commonwealth Office in London even highlights the issue in its travel advice for Greece.
This month, drivers faced a new highway code, with fines of up to €700 (£490) for ignoring a stop sign or running through a red light.
The adverse effects of billboards have been highlighted due to the efforts of an Athenian lawyer, Athanasios Tsiokos, who killed his son when he crashed into a billboard on a busy avenue in the capital. He has since campaigned to punish advertising companies, and this year his complaint was upheld by the State Council, which ordered the billboards removed.
Municipalities have begun dismantling them. “This is an issue of public safety and it only happens in our country,” said Aris Stathakis, MP for the ruling New Democracy party. “All the dangerous advertising billboards have to be removed.”
The campaign has not been easy. Corrupt local government officials have long ensured that billboards have flourished. Recently, campaigners have woken up to find that those removed frequently have been re-erected overnight.

domain: news


text:
A new design for a home temple in California has the potential to make a significant difference in the way people experience the temple experience.
The design by architect Michael Schoen, based on the design of the temple in a Chinese temple, is being developed by the Temple Design Institute.
The temple, a modern and unique design, is designed for two main functions: worship and the reception of guests.
In a temple, worship and reception take place simultaneously, as the temple provides services to the public and to the temple itself.
This temple design, called the Temple at Sea, is also designed to provide a safe environment for people to be able to pray, sing, and interact with one another.
The Temple at Landscape, a similar design, was designed to serve a more casual worship service.
The idea is to create a temple where people can come together and relax in the midst of a busy, noisy city.
“We were looking at the architecture of the Temple in the temple, and we knew we had a temple design that would be a great fit,” said Michael Schön, president and CEO of the Temples at Sea Foundation, a nonprofit organization that promotes the temple design in the United States.
“This temple design is very much in the tradition of Chinese temples.
It’s a temple with an open space in the center and a very welcoming, family-friendly atmosphere.”
Schoen’s Temple at the Sea design is based on a temple that is in the heart of the city of San Diego, California.
In this temple, the design focuses on the open spaces and welcoming nature of the design, and it has a space for guests to gather.
“It’s an open-plan temple,” Schoen said.
“You can’t put a temple on top of a mountain, but you can put a small temple in the middle of a residential neighborhood.”
A view of the original Temple at sea design at a Chinese Temple site in Hong Kong.
Courtesy of Michael Schoonsten.
“What we found is that the Temple At Sea design, for one, has the capacity to be more welcoming than a traditional temple,” he continued.
“I think it’s a very beautiful design that has the ability to be very inviting to the people who are coming to the site, because it’s an urban temple.”
The design also has the flexibility to accommodate a wide range of worship needs, including a temple designed for family gatherings, a temple for a small group of people, and a temple as a community service.
Schoen also said the temple will be able be built to accommodate the needs of a larger congregation, which can be a challenge.
“The Temple At the Sea is the first design we’ve done that has a large, open space that accommodates worship and a large community gathering,” Schön said.
The original temple in China was built to honor the Chinese emperor and is known as the Great Hall of Heaven.
The modern temple was designed by the temple’s architect, Huayong Liu, who has also designed other temples around the world.
Temple design has become a trend in the U.S. in recent years, with the popularity of temples in the cities of Los Angeles, Chicago, New York, and Washington, D.C. There are also many temples designed for weddings, funerals, and other ceremonies, and temples have become popular in schools and other residential settings.
Schön and Temple at The Sea are currently working on designing another temple in Los Angeles.
“Our first temple in New York was designed for a group of 10 people,” Schonen said.
Schoonster said he hopes to complete his temple design soon, which he believes will provide a “very welcome space.”
Temple design is not just about creating a temple in which people can gather together, but it’s also about creating an experience where people are welcome and valued.
“People are often not welcomed in temple design because of the culture they’re coming from,” he said.
Temple designers have always been able to incorporate elements of their designs into other parts of the building, such as the design that connects the temple to the rest of the structure, the shape of the floor, and the design and placement of the windows and doorways.
The more diverse the temple is, the more appropriate it is to incorporate cultural elements, said Schön.
“So, the temple designs that are used in temple sites are always going to be in the cultural context,” he added.
“But the temple that’s built in California is a very different temple than the one in New Orleans, where you have a traditional building and you have that temple design.”
A temple design should not be just a one-time project, but a series of design efforts that can be continued over time, said Temple at home designer, John C. Stearns, who is also the director of Temple Design at the Temple of Solomon Institute.
“Temple design is a process that takes time, so it’s good to have a long-term vision,” he explained.
“For instance, a lot

domain: artistic


text:
Welcome to our Prichard, Alabama Food Pantries and Soup Kitchens. Below are all of the Emergency Food Programs provided through Food Pantries and Soup Kitchens in Prichard, AL and surrounding cities. If you are searching for Prichard Food Banks - Food banks are distribution hubs. They supply the food to the Soup Kitchens, Food Pantries, Shelters etc. They in turn provide that food to the individuals that need it. Food Banks do not directly serve individuals in need.
There are no listing located within the Prichard city limits.
Free Community Soup Kitchen Every Thursday 2 - 4 p.m. Free Food Pantry (Food Boxes) Every Wednesday 10:30 a.m. - 12:30 p.m. Free Clothing Ministry - Every Wednesday and Friday 10:30 a.m. - 12:30 p. [...]
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When shorts are too short, they’re just uncomfortable. That’s where the women’s Burton Mid Short comes in. Made from a movement-encouraging stretchy blend of 99% cotton and 1% spandex, and with a 5” inseam, they are the perfect length for actually doing stuff like biking and hiking. And that’s what you’re all about, right?
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Whopping 50 percent of 18-to-24-year-olds voted in that 1972 election, according to American Demographics magazine. Voting was as cool as protesting wars in patchouli-scented scarves and drag-racing for pinks. But then something happened.
I am a 24-year-old teacher -- smack in the middle of America's "young generation" of 18-30-year-olds -- and I am troubled that half of my cohort -- my peers, my co-workers, my friends -- choose not to exercise their right to vote.
The voting power of young adults certainly exists; the issue has now just become a question of action. As a person who is both a youth voter and a college student, the non-empirically based answer seems quite easy: convenience.
Smart politicians know that the 2010 electorate is unlikely to repeat itself in 2012. Expect them to adjust their policies and rhetoric accordingly if they plan to be reelected to another term in office.


text:
Effect of tempe-yellow maize porridge and milk-yellow maize porridge on growth rate, diarrhoea and duration of rehabilitation of malnourished children.
MetadataShow full item record
The diet for 117 protein-energy-malnourished children admitted into the Mbooni Family Life Training Centre between November 1992 and March 1993, was supplemented with either tempe-yellow maize porridge (TYMP) or milk-yellow maize porridge (MYMP). Fifty-six malnourished children had their diet supplemented with TYMP. Another group of 61 children had theirs supplemented with MYMP. The growth rate (weight gain), duration of diarrhoeal episodes and rehabilitation period for each child was recorded and a comparison made between the two dietary groups. The TYMP group achieved a significantly (p < 0.05) higher growth rate (5.2 gm/kg body weight/day) compared to the MYMP whose growth rate was 2.2 gm/kg body weight/day. The mean daily protein and caloric consumption (per kg body weight) by the children in both groups (at 3.4 gm of proteins and 83 calories for the MYMP group and 3.9 gm of protein and 101 calories for the TYMP group) were not significantly different. The duration of diarrhoeal episodes for the MYMP group was significantly (P < 0.05) longer (4.6 diarrhoea days) than that of the TYMP group whose diarrhoeal episodes lasted 0.7 diarrhoea days. The rehabilitation period for the TYMP group (15 days) was significantly (p > 0.05) lower than that for the MYMP group which was 20 days. Furthermore, the cost of supplementing the diet for each child with the tempe-yellow maize porridge (KSh. 1625) was approximately 25% lower than that of supplementing it with milk-yellow maize porridge which was KSh. 2060. These results suggest that it may be more beneficial in terms of duration of both diarrhoeal episodes and rehabilitation period and overall institutional cost if malnourished children's diets are supplemented with tempe-yellow maize porridge.

domain: literature


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The first official book on the most important Italian building by the Brazilian architect Oscar Niemeyer, one of the leaders of modern architecture.
With a step-by-step study, the volume tells the story of the construction, near the city of Milan, of Niemeyer’s building, now headquarters of Mondadori Editorial Group. A collection of never-before-seen architectural plans, original drawings, and unpublished sketches offers a fascinating look at the architect’s work and a tool for all who wish to deepen their knowledge of Oscar Niemeyer’s works.
The volume is enlivened by an extraordinary photographic shoot by Roland Halbe, an international architectural photographer who captures every detail that makes this spectacular building a harmonious set and an example of indissoluble integration between form and structure, still considered one of the most unique and elegant works ever made by the Brazilian architect.
Electa has been an acknowledged leader in the various sectors of the visual arts for over 60 years, thanks to our innovative publishing initiatives, academic rigour, signature graphics and top quality illustrations which have helped build the history of illustrated publishing.
Electa is the leading player in the visual arts publishing sector in Italy with our huge catalogue and broad range of series, but also thanks to the sheer quality of our publications and our ongoing work with museums and galleries in a variety of fields.
011 Form and Strcture: Oscar Niemeyer and the Mondadori Building
035 The Project, the Construction, the Building
140 Writings about Oscar Niemeyer and the Mondadori Building


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text:
I know I want to marry you. But I wish we were the first to ever get married. I cannot help feeling that the institution has been somewhat corrupted and corroded by the misuse of others. We could show them, by a beyootiful and myoochooal respect for each other, how things must be conducted. Have I ever told you how much I love you? Well, darling, I am telling you now...
Fear, jealousy and anger are manifestations of a lack of being in love. Being in love is possible only when you are in love with yourslef first. Self-love is an inborn quality that has been corroded because of social conditioning. As a result, we forget that love is within is, or rather, we are the source of love.
The weathered dairy barn, the wilted chicken coop, the leaning corn crib, the corroded silos- all were revealed as structures of utility and grace. Someone must have rigged Ry's perception so that he had spent his whole life seeing only the ultimate futility of these structures while concealing what made them worthy, the struggle itself, the striving for a better day.
What is the Absolute? Something that appears to us in fleeting experiences--say, through the gentle smile of a beautiful woman, or even through the warm caring smile of a person who may otherwise seem ugly and rude. In such miraculous but extremely fragile moments, another dimension transpires through our reality. As such, the Absolute is easily corroded;it slips all too easily through our fingers and must be handled as carefully as a butterfly
We are living in a world of fear. The life of man today is corroded and made bitter by fear: fear of the future, fear of the hydrogen bomb, fear of ideologies. Perhaps this fear is a greater danger than the danger itself because it is fear, which drives men to act thoughtlessly, to act dangerously.
All cultures forged by nations""the noble indigenous past of America, the brilliant civilization of Europe, the wise history of Asian nations, and the ancestral wealth of Africa and Oceania""are corroded by the American way of life. In this way, neoliberalism imposes the destruction of nations and groups of nations in order to reconstruct them according to a single model. This is a planetary war, of the worst and cruelest kind, waged against humanity.
We are living in a world of fear. The life of man today is corroded and made bitter by fear-fear of the future, fear of the hydrogen bomb, fear of ideologies. Perhaps this fear is a greater danger than the danger itself, because it is fear which drives men to act foolishly to act thoughtlessly, to act dangerously.
The 16th Amendment corroded the American concept of natural rights; ultimately reduced the American citizen to a status of subject, so much so that he is not aware of it; enhanced Executive power to the point of reducing Congress to innocuity; and enabled the central government to bribe the states, once independent units, into subservience. No kingship in the history of the world ever exercised more power than our Presidency, or had more of the people's wealth at its disposal.
Of every ten who tread the Middle Way to Knowledge there are nine who turn aside through avarice, though not all avarice is born of belly-hunger or the greed for gold. Some seek preeminence, such eminence as they have won corroded by insane pride. So by this mark you shall know the Middle Way, that whoso treads it truly avoids vices, having found them in himself, so that he knows their habit and is temperate in judgment, throwing no stones lest he break the windows of his own soul.-From The Book Of The Sayings of Tsiang Samdup
Do jungle animals understand the true nature of the trees among which they have their daily being? In the parent-forest, amid those mighty trunks, we shelter and play; but whether the trees are healthy or corroded, whether they harbour demons or good spirites, we cannot say. Nor do we know the greatest secret of all: that one day we, too, will become as arboreal as they. And the trees, whose leaves we eat, whose bark we gnaw, remember sadly that they were animals once, they climbed like squirrels and bounded like deer, until one day they paused, and their legs grew down into the earth and stuck there, spreading, and vegetation sprouted from their swaying heads. They remember this as a fact; but the lived reality of their fauna years, the how-it-felt of that chaotic freedom is beyond recapture. They remember it as a rustle in their leaves.
to cheat your way into a job bigger than your mind can handle is to become a fear-corroded ape on borrowed motions and borrowed time, and to settle down into a job that requires less than your mind's full capacity is to cut your motor and sentence yourself to another kind of motion: decay - that your work is the process of achieving your values, and to lose your ambition for values is to lose your ambition to live - that your body is a machine, but your mind is its driver, and you must drive as far as your mind will take you, with achievement as the goal of your road - that the man who has no purpose is a machine that coasts downhill at the mercy of any boulder to crash in the first chance ditch, that the man who stifles his mind is a stalled machine slowly going to rust, that the man who lets a leader prescribe his course is a wreck being towed to the scrap heap, and the man who makes another man his goal is a hitchhiker no driver should ever pick up - that your work is the purpose of your life, and you must speed past any killer who assumes the right to stop you, that any value you might find outside your work, any other loyalty or love, can be only travelers you choose to share your journey and must be travelers going on their own power in the same direction.
I took a glass retort, capable of containing eight ounces of water, and distilled fuming spirit of nitre according to the usual method. In the beginning the acid passed over red, then it became colourless, and lastly again all red: no sooner did this happen, then I took away the receiver; and tied to the mouth of the retort a bladder emptied of air, which I had moistened in its inside with milk of lime lac calcis, (i.e. lime-water, containing more quicklime than water can dissolve) to prevent its being corroded by the acid. Then I continued the distillation, and the bladder gradually expanded. Here-upon I left every thing to cool, tied up the bladder, and took it off from the mouth of the retort.- I filled a ten-ounce glass with this air and put a small burning candle into it; when immediately the candle burnt with a large flame, of so vivid a light that it dazzled the eyes. I mixed one part of this air with three parts of air, wherein fire would not burn; and this mixture afforded air, in every respect familiar to the common sort. Since this air is absolutely necessary for the generation of fire, and makes about one-third of our common air, I shall henceforth, for shortness sake call it empyreal air, [literally fire-air] the air which is unserviceable for the fiery phenomenon, and which makes abut two-thirds of common air, I shall for the future call foul air [literally corrupted air].
Carl Wilhelm Scheele
Productiveness is your acceptance of morality, your recognition of the fact that you choose to live-that productive work is the process by which man's consciousness controls his existence, a constant process of acquiring knowledge and shaping matter to fit one's purpose, of translating an idea into physical form, of remaking the earth in the image of one's values-that all work is creative work if done by a thinking mind, and no work is creative if done by a blank who repeats in uncritical stupor a routine he has learned from others-that your work is yours to choose, and the choice is as wide as your mind, that nothing more is possible to you and nothing less is human-that to cheat your way into a job bigger than your mind can handle is to become a fear-corroded ape on borrowed motions and borrowed time, and to settle down into a job that requires less than your mind's full capacity is to cut your motor and sentence yourself to another kind of motion: decay-that your work is the process of achieving your values, and to lose your ambition for values is to lose your ambition to live-that your body is a machine, but your mind is its driver, and you must drive as far as your mind will take you, with achievement as the goal of your road-that the man who has no purpose is a machine that coasts downhill at the mercy of any boulder to crash in the first chance ditch, that the man who stifles his mind is a stalled machine slowly going to rust, that the man who lets a leader prescribe his course is a wreck being towed to the scrap heap, and the man who makes another man his goal is a hitchhiker no driver should ever pick up-that your work is the purpose of your life, and you must speed past any killer who assumes the right to stop you, that any value you might find outside your work, any other loyalty or love, can be only travelers you choose to share your journey and must be travelers going on their own power in the same direction.
Old Hubert must have had a premonition of his squalid demise. In October he said to me, 'Forty-two years I've had this place. I'd really like to go back home, but I ain't got the energy since my old girl died. And I can't sell it the way it is now. But anyway before I hang my hat up I'd be curious to know what's in that third cellar of mine.' The third cellar has been walled up by order of the civil defence authorities after the floods of 1910. A double barrier of cemented bricks prevents the rising waters from invading the upper floors when flooding occurs. In the event of storms or blocked drains, the cellar acts as a regulatory overflow. The weather was fine: no risk of drowning or any sudden emergency. There were five of us: Hubert, Gerard the painter, two regulars and myself. Old Marteau, the local builder, was upstairs with his gear, ready to repair the damage. We made a hole. Our exploration took us sixty metres down a laboriously-faced vaulted corridor (it must have been an old thoroughfare). We were wading through a disgusting sludge. At the far end, an impassable barrier of iron bars. The corridor continued beyond it, plunging downwards. In short, it was a kind of drain-trap. That's all. Nothing else. Disappointed, we retraced our steps. Old Hubert scanned the walls with his electric torch. Look! An opening. No, an alcove, with some wooden object that looks like a black statuette. I pick the thing up: it's easily removable. I stick it under my arm. I told Hubert, 'It's of no interest... ' and kept this treasure for myself. I gazed at it for hours on end, in private. So my deductions, my hunches were not mistaken: the Bie¨vre-Seine confluence was once the site where sorcerers and satanists must surely have gathered. And this kind of primitive magic, which the blacks of Central Africa practise today, was known here several centuries ago. The statuette had miraculously survived the onslaught of time: the well-known virtues of the waters of the Bie¨vre, so rich in tannin, had protected the wood from rotting, actually hardened, almost fossilized it. The object answered a purpose that was anything but aesthetic. Crudely carved, probably from heart of oak. The legs were slightly set apart, the arms detached from the body. No indication of gender. Four nails set in a triangle were planted in its chest. Two of them, corroded with rust, broke off at the wood's surface all on their own. There was a spike sunk in each eye. The skull, like a salt cellar, had twenty-four holes in which little tufts of brown hair had been planted, fixed in place with wax, of which there were still some vestiges. I've kept quiet about my find. I'm biding my time.


text:
Some individuals, including those who do not eat fish and those who’re at significant risk for cardiovascular events, may benefit from fish oil supplements.
More often than not, supplements are needed to achieve the omega 3 fatty acids dosage.
Fish oil, which is high in omega 3 essential fatty acids, is the most appropriate supplement to be used for this objective.
Fish oil supplements provide a good source of omega 3 essential fatty acids.
In the following paragraphs are likely health benefits research has found for fish oil or fish when added to the diet.
Many of the health benefits fish oil offers come from the omega 3 it contains.
These fatty acids which have been known to play a role in improved health, include for example, Alpha-linolenic acid, EPA (Eicosapentaenoic acid), DHA (Docosahexaenoic acid), and Gamma-linolenic acid.
Foods are simply the only supply of these omega 3 essential fatty acids, given that the body is not able to create them by itself.
Various skin disorders, like eczema and psoriasis, affect people from all over the globe, and a lot experience these disorders.
While pharmacies supply a wide range of solutions for these problems, fish oil may help improve the appearance of your skin significantly.
It’s because the omega 3 from fish oil may assist with maintaining moisture in the skin.
Dry skin or any condition wherein dry skin can become a root cause or make problems worse, such as eczema, may markedly gain from fish oil supplements.
You may add fish oil to psoriasis spots directly, working with it as a topical solution.
Research conducted in Denmark of about 9,000 women that are pregnant revealed that expecting women who took fish oil all through the pregnancy were a lot less prone to suffer from post-natal depression.
The possibility of premature labor occurring had also been substantially reduced when a woman had sufficient levels of omega 3 in her system, the same research showed.
Moreover, young ones whose mothers had taken fish oil, displayed greater hand-eye coordination unlike those who had not enjoyed the benefits of omega 3.
It must be said that a expectant woman should refrain from using supplements sourced from liver oil, such as cod-liver oil.
Pure fish oil supplements is really what one should be taking.
It’s long been understood that persons with diabetes type 2 are prone to encounter other health issues associated with this condition.
It is because incessantly high blood sugar levels can result in damage to different internal organs elsewhere in the body with time.
Diabetic patients can suffer from injury to both the large and small blood vessels, which may increase the risks of experiencing heart attacks or stroke.
Numerous researchers accept that some individuals with diabetes could notice a decrease in symptoms through closely watching dieting and exercise.
However, research from the University of Oxford in the United Kingdom revealed that diabetics receiving fish oil, may notice that blood triglyceride levels can be decreased.
Helping to keep triglyceride down, is a big factor in decreasing the risk of cholesterol-related clogged arteries, which can play a part in heart attack and stroke.
Research undertaken at the Linus Pauling Institute reveals that someone with insufficient levels of omega 3 fatty acids is significantly more likely to suffer from heart disease.
Actually, clinical tests conducted by the American Heart Association reveal that getting more omega 3 has a noticeable impact on reducing the chance of cardiovascular disease.
The research also indicates that fish oil can help to prevent the harmful build-up of triglycerides, as well as decreasing and minimizing the risk of atherosclerosis.
The study also discovered that those subjects who took fish oil supplements as part of a regular routine were far less likely to experience a fatal cardiac arrest.
A number of serious health conditions, such as heart disease, bronchial asthma, some types of cancer, rheumatoid arthritis, crohn’s disease and many more can be caused by inflammation.
The body has its own defense system against inflammation in the form of hormones called prostaglandins, whose production has been shown to be stimulated by fish oil.
Omega 3 has powerful anti-inflammatory properties, which helps to reduce inflammation in the joints caused by arthritis.
People who take fish oil supplements on a regular basis are less likely to suffer from ailments and issues triggered by inflammation.
Research conducted from the Case Western Reserve University, discovered that fish oil can be quite effective at controlling depression and anxiety.
Experts noted that the degree of depression reported in places where fish is a mainstay is significantly lower.
On the flip side, there’ve been an increasing number of instances of depression and anxiety recorded in locations where fish is not a significant part of the diet.
It was learned that symptoms of depression can be taken care of with the assistance of omega 3 fatty acids in fish oil, and it was capable of regulating stress and anxiety.
For some time now, improving blood circulation was accomplished with the help of omega 3 essential fatty acids.
It aids to keep blood pressure in check in some individuals, and reduce the odds of a cardiac arrest or stroke.
Fish oil may prove helpful in lowering the amount of triglycerides in the blood, as researchers at the University of Oxford in the UK have discovered.
Restricted blood flow due to the constriction of the arteries, is usually the result of these unhealthy fats, that are linked to cholesterol.
The brain receives more blood if your whole blood flow improves.
Anxiety and depression, and other neurological disorders, benefit from higher and balanced amount of key nutrients to the brain, due to the fact the seriousness relating to the symptoms is greatly reduced.
Asthma is a ailment that affects the airways, causing it to be challenging for individuals to breathe well.
There aren’t many of us who are conscious of just how serious long-term, chronic swelling associated with the airways is.
Still research carried out by the University of Sydney may have found out that fish oil may actively assist to lower the risk of asthma attacks.
Research undertaken in Wyoming supported these results as it was found that a number of asthma patients underwent an alleviation of their symptoms when they began to take fish oil regularly.
The minute someone suffers from an asthma attack, the airways constrict as a result of inflammation and fish oil appears to decrease the level of inflammation, as reported by French researchers.
It was also found out that overall lung function did actually improve once people started to take good levels of fish oil.
Fish oil may be able to decrease the chances of symptoms but that doesn’t indicate that you need to stop taking the treatment your physician recommended if you’re an asthmatic.
The fact that fish oil has become much more widespread and highly regarded should be abundantly clear, seeing as it offers so many benefits to anyone’s overall health.
Including certain foods into your diet two or three times each week, you will discover that it’s easy to acquire more omega 3.
you will receive the most potent amounts of omega 3 essential fatty acids from fish, but it’s also wise to include other foods too, such as eggs and nuts.
However, to guarantee your body is getting enough omega 3 essential fatty acids, you should consider fish oil supplements.
Always choose a brand that creates its supplements using good quality, pure fish oil that is contaminant-free.
Always take time to read the label so you can avoid any supplements using low quality fish oil, seeing that pure fish oil is best.

domain: biology


text:
Friday, December 2, 2011
What you need:
some wool, and
a piece of hessian.
It also helps if you have patience and quite a bit of determination!
We chose an animal that we saw at Mt Bruce and drew it onto card and cut it out. These animals were traced onto hessian. Then we had to sew around the trace mark. Sewing was very tricky. Even threading the wool into the needle was tricky but some of us could do it by ourselves.
It took a few weeks to finish these. We think they look pretty fabulous. Come into our room and see them with your own eyes.

domain: design


text:
• The Magic of Children in the Arts will be hosting FREE art workshops at the Collingwood Public Library, 55 Ste. Marie Street, Collingwood
• The workshops are for children ages 4 to 13, and will be held on:
• Saturday, February 9th From 10am to NOON and 1 pm to 3 pm
• All of the art created by the kids will be entered into this season’s Magic of Children in the Arts art show!
• Sign up at the library’s front desk, or visit: www.MagicOfChildren.ca
(Source and Image: Collingwood Public Library)

domain: entertainment


text:
Antitumor effect of iRGD-modified liposomes containing conjugated linoleic acid–paclitaxel (CLA-PTX) on B16-F10 melanoma
Received 7 April 2014
Accepted for publication 6 May 2014
Published 24 June 2014 Volume 2014:9(1) Pages 3091—3105
Checked for plagiarism Yes
Review by Single anonymous peer review
Peer reviewer comments 4
Ruo Du,1 Ting Zhong,1 Wei-Qiang Zhang,1 Ping Song,1 Wen-Ding Song,1 Yang Zhao,1 Chao-Wang,1 Yi-Qun Tang,3 Xuan Zhang,1,2 Qiang Zhang1,2
1Department of Pharmaceutics, 2State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 3Department of Clinical Pharmacy, China Pharmaceutical University, Nanjing, People’s Republic of China
Abstract: In the present study, we prepared a novel delivery system of iRGD (CRGDK/RGPD/EC)-modified sterically stabilized liposomes (SSLs) containing conjugated linoleic acid–paclitaxel (CLA-PTX). The anti-tumor effect of iRGD-SSL-CLA-PTX was investigated on B16-F10 melanoma in vitro and in vivo. The in vitro targeting effect of iRGD-modified SSLs was investigated in a real-time confocal microscopic analysis experiment. An endocytosis-inhibition assay was used to evaluate the endocytosis pathways of the iRGD-modified SSLs. In addition, the in vitro cellular uptake and in vitro cytotoxicity of iRGD-SSL-CLA-PTX were evaluated in B16-F10 melanoma cells. In vivo biodistribution and in vivo antitumor effects of iRGD-SSL-CLA-PTX were investigated in B16-F10 tumor-bearing mice. The induction of apoptosis by iRGD-SSL-CLA-PTX was evaluated in tumor-tissue sections. Real-time confocal microscopic analysis results indicated that the iRGD-modified SSLs internalized into B16-F10 cells faster than SSLs. The identified endocytosis pathway of iRGD-modified SSLs indicated that energy- and lipid raft-mediated endocytosis played a key role in the liposomes’ cellular uptake. The results of the cellular uptake experiment indicated that the increased cellular uptake of CLA-PTX in the iRGD-SSL-CLA-PTX-treated group was 1.9-, 2.4-, or 2.1-fold compared with that in the CLA-PTX group after a 2-, 4-, or 6-hour incubation, respectively. In the biodistribution test, the CLA-PTX level in tumor tissues from iRGD-SSL-CLA-PTX-treated mice at 1 hour (1.84±0.17 µg/g) and 4 hours (1.17±0.28 µg/g) was 2.3- and 2.0-fold higher than that of CLA-PTX solution at 1 hour (0.79±0.06 µg/g) and 4 hours (0.58±0.04 µg/g). The value of the area under the curve for the first 24 hours in the tumors of iRGD-SSL-CLA-PTX-treated mice was significantly higher than that in the SSL-CLA-PTX and CLA-PTX solution-treated groups (P<0.01). The in vivo antitumor results indicated that iRGD-SSL-CLA-PTX significantly inhibited the growth of B16-F10 tumors compared with the SSL-CLA-PTX or CLA-PTX solution-treatment groups (P<0.01). The results of tumor-cell apoptosis showed that tumors from the iRGD-SSL-CLA-PTX-treated group exhibited more advanced cell apoptosis compared with the control, CLA-PTX solution-, and SSL-CLA-PTX-treated groups. In conclusion, the antitumor effect of iRGD-SSL-CLA-PTX was confirmed on B16-F10 melanoma in vitro and in vivo.
Keywords: peptide-modified liposome, iRGD, CLA-PTX, antitumor effect, in vitro, in vivo
Paclitaxel (PTX) is a chemotherapeutic antitumor drug widely used in clinical situations for the treatment of many tumors.1 In order to overcome the side effects of Cremophor® EL (CrEL; BASF, Ludwigshafen, Germany), which is used as a solubilizer in the current PTX-injection formulation named Taxol® (Bristol-Myers Squibb, New York, NY, USA), many CrEL-free PTX-delivery systems have been investigated, such as liposomes, nanoparticles, and micelles.2 These PTX-delivery systems express passive targeting effects because of an enhanced-permeability-and-retention effect in tumor tissues.3,4
Conversion of PTX into a water-soluble conjugate is an important alternative strategy.5–8 Examples of this include the poly(L-glutamic acid)–PTX conjugate (coded as CT-2103), polyethylene glycol (PEG)–PTX conjugate, hyperbranched poly(ether-ester)-PTX conjugate, poly(L-γ-glutamyl-glutamine)–PTX conjugate, N-(2-hydroxypropyl)-methacrylamide–PTX conjugate, PEG–polylactic acid–PTX, and hyaluronic acid–PTX conjugate.9–16 These polymer–PTX conjugates possess many evident advantages compared to conventional polymeric carrier drug-delivery systems, including enhanced water-solubility and antitumor activities.
Another strategy for PTX conversion is to improve its lipophilicity.17–19 Modification of PTX with fatty acid derivatives is one potential approach. It has been reported that docosahexaenoic acid–PTX is currently in Phase III clinical trials.20 According to this strategy, we previously reported a new PTX–fatty acid-derivative conjugate, involving the conjugation of linoleic acid with PTX (CLA-PTX).21 The antitumor effect of CLA-PTX in C6 glioma and B16-F10 melanoma cells was confirmed by its in vitro cytotoxicity.21–23 CLA-PTX also exhibits higher cellular uptake in C6 glioma and B16-F10 melanoma cells compared with PTX. The in vivo antitumor activity of CLA-PTX was confirmed in brain tumor-bearing rats. However, the solvent used for the CLA-PTX formulation for intravenous (IV) administration was a mixture of physiological saline, CrEL, and ethanol (80:10:10, volume:volume:volume [v:v:v]), the same as that used for Taxol.21 Therefore, a novel CLA-PTX microemulsion containing less CrEL compared with the CLA-PTX solution was designed and prepared.22 The antitumor activity of the CLA-PTX microemulsion was confirmed in our in vivo C6 glioma tumor-bearing nude mouse model as well as in a rat model in the treatment of brain tumors.22 In addition, the CLA-PTX microemulsion is safer than CLA-PTX solution due to the reduction in CrEL. Therefore, we considered that the CrEL-free as well as the active targeted modified CLA-PTX-delivery system would have none of the toxicity produced by CrEL and would also exhibit increased antitumor activity.
It has been reported that the iRGD peptide (sequence CRGDK/RGPD/EC) could increase vascular and tissue permeability in a tumor-specific and neuropilin-1-dependent manner.24,25 The targeting activity of iRGD-modified sterically stabilized liposomes (SSLs) has been demonstrated in our previous flow-cytometry and confocal analysis studies in B16-F10 cells.26 However, the cellular uptake pathway of iRGD-modified SSLs in B16-F10 cells was unclear. In addition, we found that the antitumor activity of CLA-PTX solution on B16-F10 bearing mice was similar to that of PTX injection.23 Therefore, we suggested that the antitumor effect of the CLA-PTX formulation after being iRGD-modified would be increased in B16-F10 melanoma cells. This assumption needs to be confirmed in detail.
In the present study, iRGD was selected as a targeting ligand, SSLs were used as a delivery system, and CLA-PTX was selected as a model drug to prepare CLA-PTX-loaded iRGD-modified SSLs (iRGD-SSL-CLA-PTX). Because of overexpression of both αv-integrin receptors and neuropilin-1 (NRP-1), B16-F10 melanoma cells were chosen for the tumor-cell model. The antitumor effect of iRGD-SSL-CLA-PTX was investigated on B16-F10 melanoma in vitro and in vivo. The in vitro targeting effect and endocytosis pathway of iRGD-modified SSLs were investigated. In addition, the in vitro cellular uptake and in vitro cytotoxicity of iRGD-SSL-CLA-PTX were also evaluated. The in vivo biodistribution and the in vivo antitumor effect of iRGD-SSL-CLA-PTX were investigated in B16-F10 tumor-bearing mice. The induction of apoptosis by iRGD-SSL-CLA-PTX was evaluated in tumor-tissue sections.
Materials and methods
PTX was obtained from Yunnan Hande Bio-Tech (Kunming, People’s Republic of China [PRC]). CLA-PTX was synthesized from CLA and PTX according to our previous report.21 The structure of CLA-PTX is shown in Figure 1. The iRGD peptide (CRGDK/RGPD/EC) was supplied by GL Bio-Chem (Shanghai, PRC). Cholesterol, sulforhodamine B, chlorpromazine, amiloride, filipin, methyl-β-cyclodextrin (M-β-CD) and Tris base were purchased from Sigma-Aldrich (St Louis, MO, USA). Coumarin-6 and Hoechst 33258 were obtained from Thermo Fisher Scientific (Waltham, MA, USA). Egg phosphatidylcholine was purchased from Lipoid (Ludwigshafen, Germany). 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG) was obtained from Avanti Polar Lipids (Alabaster, AL, USA). iRGD-PEG-DSPE, as shown in Figure 1, was synthesized according to our previously reported method.26
Cell-culture media, Dulbecco’s Modified Eagle’s Medium DMEM (high glucose), penicillin–streptomycin, fetal bovine serum, and L-glutamine were purchased from Thermo Fisher Scientific. All solvents and other reagents were of analytical grade.
Murine B16-F10 cells (Chinese Academy of Sciences Cell Bank, Shanghai, PRC) were grown in Dulbecco’s Modified Eagle’s Medium (high glucose) supplemented with 10% fetal bovine serum and 1% glutamine penicillin–streptomycin at 37°C and 5% CO2.
Female C57BL6/N mice weighing 20–25 g (5–6 weeks old) were obtained from the Experimental Animal Center of Peking University Health Science Center (Beijing, PRC). All care and handling of the animals were performed according to the requirements of the Institutional Authority for Laboratory Animal Care of Peking University. For preparation of the tumor-bearing C57BL6/N mice model, C57BL6/N mice received a subcutaneous inoculation of 100 μL B16-F10 cell suspension (1×106) in the right armpit.
Preparation of iRGD-SSL-CLA-PTX
The iRGD-modified SSLs containing CLA-PTX were prepared as described previously.27 As shown in Figure 1, a mixture of egg phosphatidylcholine, cholesterol, PEG-DSPE, and iRGD-PEG-DSPE (65:30:4:1 molar ratio) was dissolved in chloroform, then CLA-PTX (CLA-PTX:lipid [weight:weight] ratio of 1:20) was added, and the resulting solution was then dried to obtain the lipid film. The resulting thin film was hydrated in phosphate buffered saline (PBS; pH 7.4) and sonicated to produce a liposome suspension. After that, the liposome suspension was passed through a 0.2 μm pore-size membrane, then through a Sephadex G-50 column to remove any free CLA-PTX, obtaining the final iRGD-SSL-CLA-PTX.
To prepare the SSL-CLA-PTX, an identical procedure was performed, except that the equivalent molar iRGD-PEG-DSPE was replaced by PEG-DSPE. The SSL-CLA-PTX was used as nontargeting liposome. Coumarin-6-loaded liposomes (iRGD-SSL-coumarin-6 and SSL-coumarin-6) were prepared exactly the same as for CLA-PTX-loaded liposomes, except that CLA-PTX was replaced by coumarin-6. To prepare the CLA-PTX solution, CLA-PTX was directly dissolved in a mixture of physiological saline, CrEL, and ethanol (80:10:10, v:v:v).
Characterization of iRGD-SSL-CLA-PTX
The particle size and zeta potential of iRGD-SSL-CLA-PTX (0.3 mg/mL) were measured by a Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK) at 25°C. For measurement of the encapsulation efficiency of iRGD-SSL-CLA-PTX, the final liposomes were passed through the Sephadex G-50 column to remove free CLA-PTX, followed by disruption with 10% Triton X-100 (v:v), then the CLA-PTX in the liposomes was measured by high-performance liquid chromatography (HPLC). Meanwhile, the same amount of iRGD-SSL-CLA-PTX was treated as previously described, except that it was not passed through a Sephadex G-50 column to obtain the total concentration of CLA-PTX. The encapsulation efficiency was calculated using the formula below:
In vitro release of CLA-PTX from iRGD-SSL-CLA-PTX
The CLA-PTX released from iRGD-SSL-CLA-PTX was investigated by dialysis under sink conditions. Briefly, a volume of 1 mL iRGD-SSL-CLA-PTX (0.15 mg/mL) was put in a dialysis bag (molecular weight cutoff 8,000–14,000). The dialysis bag was immersed in 40 mL release medium (distilled water containing 0.1% [v:v] Tween 80) and incubated in an orbital shaker at 37°C with mild oscillating at 100 rpm for 48 hours. Each sample (0.5 mL) was taken from the release medium at a predetermined time and replaced with an equal volume of fresh release medium. After the last sample was removed at 48 hours, the solution in the dialysis bag was mixed with the release medium. Then, 10 μL of 10% (v:v) Triton X-100 was added to the release medium, and this mixture was used as a positive control. The concentration of CLA-PTX was determined by HPLC, as described in a later section.
Cellular uptake pathway of iRGD-modified liposomes
To evaluate the effect of inhibitors on the cellular uptake of iRGD-modified SSLs, B16-F10 cells were preincubated with different inhibitors for 30 minutes at 37°C. Briefly, the cells were incubated for 24 hours, then chlorpromazine (10 μg/mL), amiloride (20 μM), filipin (5 μg/mL), or M-β-CD (2.5 mM) was added, followed by incubation for 30 minutes at 37°C. After that, the inhibitor-containing culture medium was replaced with iRGD-SSL-coumarin-6 (the final concentration of coumarin-6 was 150 ng/mL) and incubated for 2 hours at 37°C. Then, the cells were washed three times with PBS solution, harvested by trypsinization, and centrifuged at 1,000 rpm for 5 minutes. After being resuspended in 500 μL PBS medium, the cells were tested using a FACScan™ (BD Biosciences, San Jose, CA, USA). The coumarin-6 in the cells was excited with an argon laser (467 nm), and fluorescence was detected at 502 nm. To investigate the effect of temperature on cellular uptake, the cells without inhibitor treatment were incubated at both 37°C and 4°C, and treated as described earlier.
Real-time confocal microscopic analysis
To study the real-time cellular uptake of iRGD-SSL-coumarin-6, B16-F10 cells (3×105 per dish) were seeded on glass-bottom dishes and allowed to grow and differentiate for 4 days prior to being studied. After that, the medium was removed, and each dish was placed on a microscope stage. Each solution containing SSL-coumarin-6, iRGD-SSL-coumarin-6, or coumarin-6 solution (final drug concentration of coumarin-6, 150 ng/mL) was added to each dish, respectively. The series of images were recorded continuously using a TCS SP2 confocal laser-scanning microscope (Leica Microsystems, Wetzlar, Germany). For quantification of the uptake rate, regions of interest (ROIs) were randomly selected from the cell layer, and their fluorescence intensity was determined using the SP2 confocal software. Each spot on the curve was calculated using the fluorescence intensity of 20 ROIs.
In vitro cellular uptake of iRGD-SSL-CLA-PTX
B16-F10 cells (1×106 cells/well) were seeded in six-well flat-bottom tissue-culture plates for a 24-hour incubation. After that, the medium was replaced by iRGD-SSL-CLA-PTX, SSL-CLA-PTX, or CLA-PTX solution (10 μM), and incubated for 2, 4, or 6 hours at 37°C. After incubation, cells were washed twice with cold PBS and collected using 0.25% trypsin. The cells were centrifuged at 6,000 g for 5 minutes, and the supernatant removed carefully. The cell pellets were added to 0.1 mL 10% sodium dodecyl sulfate to destroy the cells. A volume of 0.1 mL acetonitrile was added to each cell solution to precipitate the protein. After centrifugation at 6,000 g for 5 minutes, a volume of 50 μL supernatant was used for HPLC analysis. Three wells were measured for each sample. Cellular uptake efficiency was calculated using the formula below:
In vitro cytotoxicity
B16-F10 cells (5×103 cells/well) were seeded in 96-well transparent plates and incubated for 24 hours. The medium was removed, and increasing concentrations of iRGD-SSL-CLA-PTX was added. At 48 hours’ incubation, cell viability was determined by sulforhodamine B assay. Absorbance was measured at 540 nm using a 96-well plate reader (model 680; Bio-Rad Laboratories, Hercules, CA, USA). The survival percentages were calculated using the formula: survival % = (A540 nm for the treated cells/A540 nm for the control cells) ×100%, where A540 nm is the absorbance value. Each assay was carried out in triplicate. The half-maximal inhibitory concentration (IC50) values were calculated according to the dose–effect curves.
When the tumor volume reached about 300 mm3, the tumor-bearing mice were randomly assigned to one of three groups: group 1 was given IV CLA-PTX solution, group 2 was given IV SSL-CLA-PTX, and group 3 was given IV iRGD-SSL-CLA-PTX. All the CLA-PTX preparations were injected through the tail veins at a dose of 2 mg CLA-PTX/kg. After drug administration, the mice were killed at 1, 4, 8, 12, and 24 hours. The tumors or excised organs (heart, liver, spleen, lung, kidney, and brain) were collected, blotted with a paper towel, rinsed in saline, blotted to remove excess fluid, weighed, and homogenized. The homogenized tissue samples were extracted using our previously reported method.21 The content of CLA-PTX in each tissue was measured by HPLC under the chromatographic conditions described in the “HPLC analysis of CLA-PTX” section.
In vivo antitumor activity of iRGD-SSL-CLA-PTX
When the tumor volume reached about 100–150 mm3, the tumor-bearing mice were randomly assigned to one of four groups: group 1 was given IV physiological saline as a control, group 2 was given IV CLA-PTX solution, group 3 was given IV SSL-CLA-PTX, and group 4 was given IV iRGD-SSL-CLA-PTX. Each group contained six animals. All the CLA-PTX preparations were injected through the tail veins at a dose of 2 mg CLA-PTX/kg on days 7, 9, 11, 13, and 15. Throughout the study, animals were weighed and tumors measured with calipers twice a week. Tumor volumes were calculated according to the formula: V = length (cm) × width2 (cm2) ×0.5236. On day 19 of tumor inoculation, one or two mice in each group were killed and the tumors collected for the preparation of sections. Terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling (TUNEL) staining of the paraffin-embedded tumors was performed according to the standard protocols provided by the manufacturers. The survival time was calculated from the day that the B16-F10 cells were inoculated (day 0) to the day the animal died. Kaplan–Meier survival curves were drawn for each group.
HPLC analysis of CLA-PTX
The content of CLA-PTX was measured by a Waters HPLC system consisting of a 1525 pump, and a 2487 ultraviolet detector (Waters, Milford, MA, USA). The wavelength was set at 227 nm. The mobile phase was started with solvent A (acetonitrile:water 60:40 v:v) for 10 minutes, and then a linear gradient was used to change to solvent B (acetonitrile) at 12 minutes, remaining at solvent B for 20 minutes. The flow was set at 1 mL/min. An ODS 3 C-18 analytical column (5 μm, 250×4.6 mm; Phenomenex, Torrance, CA, USA) was used for chromatographic separation. The retention time of CLA-PTX was approximately 15 minutes.
All data are shown as means ± standard deviation. One-way analysis of variance was used to determine the significance of differences among groups, after which post hoc tests with the Bonferroni correction were used for comparisons among the individual groups. Statistical significance was set at P<0.05.
Characterization of iRGD-SSL-CLA-PTX
The average particle size of iRGD-SSL-CLA-PTX was about 93±0.2 nm, with a polydispersity index of 0.19±0.01, as shown in Table 1. The zeta potential of iRGD-SSL-CLA-PTX was slightly negative (−23.97±0.35 mV). The entrapment efficiency of iRGD-SSL-CLA-PTX was 83.22%, which indicated that CLA-PTX was almost completely entrapped within the liposomes. The typical particle size and distribution of iRGD-SSL-CLA-PTX is shown in Figure S1.
Table 1 Characteristics of iRGD-SSL-CLA-PTX (means ± standard deviation, n=3)
In vitro release of CLA-PTX from iRGD-SSL-CLA-PTX
The in vitro released CLA-PTX from iRGD-SSL-CLA-PTX is shown in Figure 2. The CLA-PTX released from the iRGD-SSL-CLA-PTX groups at the 48-hour time point was only 40%. Similar CLA-PTX-release behavior was observed in the SSL-CLA-PTX groups. However, CLA-PTX was rapidly released from the CLA-PTX solution, and this was almost complete within 48 hours.
Cellular uptake pathways
In order to investigate the endocytosis pathway for iRGD-modified SSLs, an endocytosis-inhibition assay was performed. As shown in Figure 3, the rate of uptake of B16-F10 cells was inhibited by 51% at 4°C compared with that at 37°C (P<0.01), and was also inhibited by M-β-CD (down to 76%) (P<0.01), showing energy- and lipid raft-mediated endocytosis. Other inhibitors, such as chlorpromazine, amiloride, and filipin, had only a weak inhibitory effect.
Real-time confocal microscopic analysis
Figure 4, A and B show that iRGD-SSL-coumarin-6 was internalized into B16-F10 cells faster and exhibited higher fluorescence intensity at every time compared with SSL-coumarin-6. In addition, coumarin-6 was also rapidly absorbed into the B16-F10 cells (Figure S2).
To further investigate the internalization speed and degree of iRGD-SSL-coumarin-6 and SSL-coumarin-6, a quantitative analysis of the images was conducted. Twenty ROIs were randomly selected to determine the fluorescence intensity. As shown in Figure 4C, iRGD-SSL-coumarin-6 showed greater internalization in B16-F10 cells at every time point compared with SSL-coumarin-6. We found that the fluorescence intensity of iRGD-SSL-coumarin-6 internalized in B16-F10 cells at the 80-second time point was almost 100% compared with that at the end of the test (Figure 4C). On the other hand, only about 30% was found in SSL-coumarin-6 groups.
In vitro cellular uptake of iRGD-SSL-CLA-PTX
The uptake of CLA-PTX by B16-F10 cells in the iRGD-SSL-CLA-PTX-, SSL-CLA-PTX-, and CLA-PTX-solution groups is shown in Figure 5. The cellular uptake of CLA-PTX in the iRGD-SSL-CLA-PTX group was significantly higher than that in the SSL-CLA-PTX- and CLA-PTX-solution groups (P<0.01) after 2, 4, and 6 hours’ incubation. These results indicated that the increased cellular uptake of CLA-PTX in the iRGD-SSL-CLA-PTX-treated group was 1.9-, 2.4-, or 2.1-fold compared with that in the CLA-PTX group after 2, 4, or 6 hours’ incubation, respectively. For SSL-CLA-PTX groups, there was 1.2-, 1.5-, or 1.3-fold increased uptake compared with that in CLA-PTX group after 2, 4, or 6 hours’ incubation, respectively.
In vitro cytotoxicity
B16-F10 cells were used to evaluate the cytotoxicity of iRGD-SSL-CLA-PTX in comparison with SSL-CLA-PTX and CLA-PTX solutions. IC50 values are given in Table 2. The IC50 value of iRGD-SSL-CLA-PTX was similar to that of CLA-PTX (P>0.05). The IC50 value of SSL-CLA-PTX was significantly higher than that of iRGD-SSL-CLA-PTX and CLA-PTX (P<0.01).
The concentration of CLA-PTX in the tumor tissues and excised organs (heart, liver, spleen, lung, kidney, and brain) of tumor-bearing mice treated with CLA-PTX solution, SSL-CLA-PTX, and iRGD-SSL-CLA-PTX is shown in Figure 6. The calculated area under the curve for hours 0–24 (AUC0–24) is shown in Table 3. The recovery efficiency of CAL-PTX in organs was in the range of 81%–94%. A targeting effect of iRGD-SSL-CLA-PTX was observed in tumor tissues compared with the SSL-CLA-PTX and CLA-PTX solutions. As shown in Figure 6, the CLA-PTX level in tumor tissues from iRGD-SSL-CLA-PTX-treated mice at 1 hour (1.84±0.17 μg/g) and 4 hours (1.17±0.28 μg/g) was 2.3- and 2.0-fold higher than that of CLA-PTX solution at 1 hour (0.79±0.06 μg/g) and 4 hours (0.58±0.04 μg/g). The AUC0–24 value in the tumors of the iRGD-SSL-CLA-PTX-treated mice was significantly higher than that in the SSL-CLA-PTX and CLA-PTX solution-treated groups. In addition, drug accumulation in the liver was higher than that in other tissues.
In vivo antitumor activity of iRGD-SSL-CLA-PTX
As shown in Figure 7, tumor growth was significantly inhibited in all CLA-PTX treatment groups compared with the physiological saline treatment group (P<0.01). iRGD-SSL-CLA-PTX significantly inhibited the growth of B16-F10 tumors compared with the SSL-CLA-PTX or CLA-PTX solution-groups (P<0.01). The average tumor size at day 19 in the CLA-PTX-solution, SSL-CLA-PTX, and iRGD-SSL-CLA-PTX groups was 4,723±853, 3,969±495 and 1,793±355 mm3, respectively, compared with 10,054±996 mm3 in the physiological saline group (P<0.01). The corresponding tumor-growth inhibition in the SSL-CLA-PTX- and iRGD-SSL-CLA-PTX-treated groups was 52.5% and 82.7%, respectively. Figure 8 shows the Kaplan–Meier survival curve. After five administrations, the median survival time of mice treated with iRGD-SSL-CLA-PTX (34 days) was significantly longer than that of mice treated with physiological saline (24 days, P<0.01), CLA-PTX solution (28 days, P<0.01), and SSL-CLA-PTX (29 days, P<0.01), respectively.
We also evaluated the effect of tumor-cell apoptosis by TUNEL analysis staining of tumor-tissue sections. As shown in Figure 9A, tumors from the iRGD-SSL-CLA-PTX-treated group exhibited more advanced cell apoptosis compared with the control, CLA-PTX solution-, and SSL-CLA-PTX-treated groups. The calculated results are shown in Figure 9B.
The in vitro cellular uptake and in vitro cytotoxicity of CLA-PTX in B16-F10 melanoma cells were demonstrated in our previous research study;23 however, the efficacy of active targeting of a modified CLA-PTX delivery system is unknown. Therefore, we designed and prepared iRGD-SSL-CLA-PTX and investigated its antitumor effect in B16-F10 melanoma cells in vitro and in vivo.
The tumor-targeting and tumor-penetrating mechanisms of iRGD have been reported.24,25 Recently, iRGD-modified delivery systems have been reported for targeted tumor therapy.28–30 Our previous results indicated that iRGD could target cells that over-express αv-integrins or the NRP-1 receptor.26 The targeting effect of iRGD-modified SSLs on B16-F10 melanoma cells has been demonstrated by our previous in vitro flow cytometry and confocal microscopy.26 In order to further confirm the targeting effect of iRGD-modified SSLs on B16-F10 melanoma cells in the present study, real-time confocal microscopic analysis was performed. The real-time uptake results indicated that iRGD-modified SSLs internalized into B16-F10 cells faster than unmodified SSLs at every time point (Figure 4). This is because ligand-modified delivery systems may prove beneficial in increasing drug exposure, due to increased target-cell uptake and target-tissue retention compared to ligand-lacking delivery systems.31 Our in vitro cellular uptake experiment also demonstrated that the iRGD-SSL-CLA-PTX was absorbed more than unmodified SSLs (Figure 5). In addition, a higher level and AUC of CLA-PTX in tumor tissue in the iRGD-SSL-CLA-PTX-treatment group was also observed in our in vivo biodistribution experiment (Figure 6). In addition, drug accumulation in the liver was higher than that in other tissues. It has been reported that iRGD-modified nanoparticles accumulate in the liver at high levels.32,33 Those researchers suggested that these nanoparticles were rapidly recognized by the phagocytic cells and the reticuloendothelial system or easily penetrated through the endothelial wall of liver fenestrae, resulting in enhanced liver uptake.
Endocytosis is a fundamental process that is used to internalize molecules and macromolecules by cells. Many nanoparticles have been customized to enter cells through endocytosis and deliver the cargo within the cells,34 such as clathrin-mediated and caveolae-mediated endocytosis, as well as rhodamine A-mediated and CDC42-mediated macropinocytosis. For example, conventional liposomes are taken up by cells via clathrin-mediated endocytosis.35,36 It has been reported that RGD-modified nanoparticles can be internalized via the receptor-mediated or clathrin-mediated endocytosis pathway.37 Therefore, understanding the cellular entry of nanoparticles has become a key factor in drug delivery. Many endocytic inhibitors have been used to characterize the endocytosis pathway involved in nanoparticles.38–42 In the present study, we selected chlorpromazine, amiloride, filipin, and M-β-CD as endocytic inhibitors to investigate the endocytosis pathway of iRGD-modified SSLs. Also, 4°C was selected to investigate the impact of energy on cellular uptake. Our results indicate that 4°C and M-β-CD significantly reduced the cellular uptake of iRGD-modified SSLs, suggesting that energy- and lipid raft-mediated endocytosis play a critical role in the cellular uptake of iRGD-modified SSLs (Figure 3). It was found that chlorpromazine, amiloride, and filipin did not reduce the cellular uptake of iRGD-modified SSLs, indicating that the clathrin-, macropinocytosis-, and caveolae-mediated endocytosis pathways are not involved in iRGD-modified SSL cellular transport (Figure 3). Our results suggest that iRGD-modified SSLs, unlike RGD-modified nanoparticles, are not internalized via the clathrin-mediated endocytosis pathway.37 Although iRGD has the RGD sequence, we speculated that the endocytosis pathway of iRGD-modified SSLs was different from that of RGD-modified nanoparticles.
It has been reported that liposomes could be efficiently accumulated in tumor because of the enhanced-permeability-and-retention effect. The addition of targeting moieties, such as antibodies or Fab fragments or small peptides and aptamers, to the surface of liposomes allows liposomes to actively target tumor cells. A lipsome is a spherical vesicle consisting of a lipid bilayer that can encapsulate various types of drugs into an inner aqueous phase or lipid bilayer.43–45 Previously, we prepared iRGD-modified SSLs containing the aqueous antitumor drug doxorubicin.26 In the present study, we prepared RGD-SSL-CLA-PTX. Our results also demonstrated the antitumor effect of iRGD-SSL-CLA-PTX (Figures 7–9).
In the present cellular uptake study, we did not observe PTX that was hydrolyzed from CLA-PTX. In addition, our biodistribution study indicated that intact CLA-PTX was identified in tumor tissue and other organs after iRGD-SSL-CLA-PTX, SSL-CLA-PTX, or CLA-PTX solution was given to tumor-bearing mice. According to these results, we suggest that the antitumor activity was due to the intact CLA-PTX, not the hydrolyzed PTX.
In the present study, we prepared a novel iRGD-SSL-CLA-PTX solution with the objective of investigating its in vitro and in vivo targeting and antitumor activity in B16-F10 cells. The targeting effect of iRGD-modified SSLs was demonstrated by the real-time confocal microscopy analysis and in vivo biodistribution experiments. The results of the endocytosis pathway of iRGD-modified SSLs indicated that energy- and lipid raft-mediated endocytosis played a key role in the cellular uptake of iRGD-modified SSLs. In addition, iRGD-SSL-CLA-PTX produced a very marked in vivo antitumor effect in B16-F10 tumor-bearing mice.
We gratefully acknowledge the financial support from the National Natural Science Foundation of China (81172992), the National Basic Research Program of China (973 Program 2013CB932501), and the Innovation Team of the Ministry of Education (BMU20110263).
The authors report no conflicts of interest in this work.
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Figure S1 Typical particle size and distribution of the SSL-CLA-PTX (A) and iRGD-SSL-CLA-PTX (B).
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domain: medical


text:
Paraffin wax is a kind of mineral wax, also is a kind of petroleum wax, mainly is C, H two kinds of elements mixed material, its
chemical formula CxHy. It is a by-product of crude oil processing and forms flake or needle-like crystals, also known as
crystalline wax. All kinds of waxed paper with good waterproof performance can be made after the paper is immersed in paraffin
wax, which can be used for food, medicine and other packaging, metal rust prevention and printing industry; Paraffin wax added to
cotton yarn can make textiles soft, smooth and elastic; Paraffin can also be detergents, emulsifiers, dispersants, plasticizers,
grease and so on.

domain: chemistry


text:
1760 – 1789
LOUIS XVI STYLE
Excavations of Herculaneum (1719) and that of Pompeii (1748) mark a return to antiquity sobriety in the Greco-Roman style. This archaeological discovery gave birth to the Louis XVI style. Mme de Pompadour (1721-1764), adheres to this “Greek” style of a new kind, it will gradually impose itself with Marie-Antoinette de Hasbourg (1755-1793), wife of Louis XVI who succeeded her as Ambassador of the Arts.
Artists and craftsmen of the Louis XVI style were able to draw inspiration from Greco-Roman art and nature with dexterity and harmony in proportions that make the Louis XVI style known and appreciated beyond national borders.
Louis XVI style is characterized by its straight lines and the symmetry of the decorations. The frames moldings are always in oak, beech, amaranth or walnut, and European linden for the sculpture.
Ornamentations are inspired by the antique, nature, exoticism: Garlands of flowers, ribbon knots, palmettes, pearls, medallions, foliages, acanthus leaves, olive leaves and laurel, wicker baskets, sickles, bee hives, pastoral, shepherdesses, country scenes, chinoiseries (asian inspiration)…
Carved woods are gilded or painted in a pastel rechampi at gold.
Louis XVI mirror will become an essential object of the decoration of its time.
Lounges and bedrooms in bourgeois and noble residences are decorated with pediment mirrors. Louis XVI style pediment mirror usually has a stylized carved wood knot. We also find pareclose mirrors and trumeau mirrors decorated with ribbons and knots.
It is under the Louis XVI style that the wedding mirrors appear which are part of the bride’s trousseau.
It was towards the end of the reign of Louis XVI that the first stucco framed mirrors appeared with carved wooden parts.
Manufacture of mercury glass mirror is still carried out in the royal manufacture which in the french revolution of 1789 will lose its privileges and monopoly.
Louis XVI style persisted until 1792 by adding revolutionary emblems to it, or was replaced by manufacture “at the Etruscan” announcing the Directoire style.


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On 9 April the Netherlands’ National Growth Fund announced to invest €615 million Euro into Quantum Delta NL, to further scale the growth of our national quantum ecosystem. Part of this investment is an amount of € 150 million for NanoLabNL to improve and expand the national nanolab facility infrastructure.
Quantum Delta is organising an online information session to inform the quantum community about the process behind the full endorsement of the proposal, further explain the proposal itself, what this means for Quantum Delta NL and what this could potentially mean for those working with quantum technology in the Netherlands.
The event will take place on Thursday 20 May and starts at 10.30 CEST.
Details on the programme and registration can be found at the website of Quantum Delta.