Många undrar om JAG ska till Göteborg Horse Show! Här får ni ert svar.
JA, jag ska till Göteborg Horse Show 2014! Jag kommer dit den 27 Februari. Kommer vara där runt 10.00 på morgonen och se på förmiddagen och sedan på på mässan!
Så om NI ska dit då, kom fram och snacka, ta bild, autograf. Vad ni vill och är nyfikna på!
Här får ni lite bilder ifrån förra årets horse show i februari.
Hej, kan du läsa min blogg?
Är du misslisibells största fan lina.
Jag ska dit på lördagen efter sport lovet. Ses kanske!
Emilia Toftebjörk. När ska du dit?
Vilken plats kommer du sitta på??
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text:
The First Snow of the Season
"He [God] giveth snow like wool: He scattereth the hoarfrost like ashes. He casteth forth His ice like morsels: who can stand before His cold? He sendeth out His word, and melteth them: He causeth His wind to blow."
~ Psalm 147:16-18 ~
s the jeep neared home, Abby glanced at her quiet passenger. She noticed that the nervous excitement in his face was starting to give way to weariness, for he could still feel the lingering effects of the hospital medications. Sedated with drugs and fatigue, the young man was becoming increasingly anxious to reach home so he could finally lay down and let the pain in his chest subside. Just as he began to think that he could wait no longer, Jake saw the small yellow house he called home, suddenly come into view.
"We're home," announced Abby, pulling the vehicle as close as she could to the front of the house. "Do you think you can make it to the front door, Jake? I'm afraid we don't have a wheelchair, like at the hospital."
"I just want to lay down," he softly breathed. Seeing Abby still looked concerned, he added, "I can make it."
John and Terry got out of the jeep and went to Jake's passenger door, ready to help in whatever way they could.
Mentally preparing himself for the pain that would follow, Jake carefully stepped out of the jeep, and weakly stood up in the bracing November air, his knees buckling ever so slightly beneath him. The ex-convict wasn't wearing the coat Abby had offered him earlier that morning, and he was beginning to regret it. The cold effortlessly penetrated his thin, long sleeved shirt, chilling him to his very core.
When Abby joined him on the other side of the vehicle, something small and white, gently floated to the ground. The snowflake was soon joined by its friend, and others quickly followed. Before long, the air was scattered with delicate bits of frosty white.
"What do you know," smiled John, looking up at the overcast sky. "It's the first snow of the season."
"Yeah," added Terry, "but it'll probably melt by tomorrow afternoon."
"Jake, let's get you inside," coaxed Abby. "I should have made you wear that coat. Your body has enough to overcome right now, without catching a cold."
Resisting help from anyone, Jake insisted on walking to the porch under his own power. Racing ahead of him, Abby unlocked the front door and swung it wide open.
"Head straight for your bedroom," she needlessly instructed Jake, walking ahead so she could turn down the blankets and make sure everything was perfectly ready for him.
Terry had been quite excited to show Jake the new fireplace and all its novelties, but he quickly realized that this was not the time. Jake was fading fast, and the sooner he went to bed, the better. Subduing his own excitement, Terry watched as John walked down the hall behind the young man, making sure that he was there to help, if called upon.
As the three men finally entered the bedroom at the end of the hall, Abby folded her arms and sighed.
"Jake," she informed him, "you can't stay in those jeans and shirt."
"But, I always sleep in day clothes," Jake weakly protested, sinking down onto the edge of his bed.
"Dad bought you some nice, long nightshirts," continued Abby, trying to sound upbeat. When Jake opened his mouth to resist, she went to his dresser and pulled out one of the white garments. "You can't bend over," she pointed out, "so this will make trips to the bathroom much easier for you. And unless you're ready for someone to help you in the bathroom, just stand in the shower when you need to clean yourself, all right?"
"If you say so," sighed Jake, for he had to admit that she was making sense. "Just let me lay down for awhile," he whispered, ready to recline that very instant. "I'm so tired, Abby."
"Oh, no you don't!" exclaimed his wife. "I want you to change into that nightshirt BEFORE those painkillers from the hospital wear off any more. Now, do you want me to help, or Dad and Uncle Terry?"
Even through his fatigue, Jake glowered at her defiantly.

redpajama_set_name: RedPajamaCommonCrawl


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Libertarian Kim Tavendale for Colorado state rep fighting to end crony capitalism
By Staff on October 5, 2016 in News
Libertarian Kim Tavendale is running for on open state rep seat in Colorado's 33rd district to "root out crony capitalism."
She proposes eliminating regulations that insulate market incumbents from competition and abolishing government subsidies of specific industries or companies.
She calls for eliminating business taxes, which, she says, "hide the cost of taxation, burden employers, and reduce business activity."
She strongly favors a repeal of Obamacare, arguing that "federal policies are a major source of the pathologies in our healthcare system."
On education, the Libertarian proposes private-school deregulation and public-school reform that would ease licensing and certification rules, accreditation requirements, and reduce state and federal mandates.
Tavendale also promises to work tirelessly to stop wasteful government spending and get government out of businesses where it does not belong.
"These days, politicians run trains, bail out banks, construct houses, sell insurance, print books, and build basketball courts," she said. "But the fact is, every service supplied by the government can be provided better and cheaper by private business in a competitive marketplace."
On drugs, she argues that drug use that does not harm others is a personal matter and "should not be punished."
Ms. Tavendale is in politics because of the arrogance of power. Not her own, but of those who hold elected office in her home state of Colorado.
She vividly remembers the day several years ago when she called the office of her state representative, Democrat Dianne Primavera, to voice her opposition to a gun bill being considered by the legislature.
Among other things, the measure provided for limiting the size of ammunition magazines, making the gun Tavendale had at home practically illegal.
She wanted to give Rep. Primavera a piece of her mind, but did not even get through.
"I spoke to someone in her office, and the essential message I got from that conversation was 'We don't care,' " Tavendale recalls. "I understood that my opinion was not even wanted."
It is this arrogance of power, which has metastasized from faraway Washington all the way to Denver and beyond, that Tavendale, a multifaith minister from Broomfield, Co., is vowing to fight.
"We have too many career politicians," she told LP News. "It is not what the Founding Fathers wanted. Ordinary people don't get involved in politics anymore. And if we don't get involved, we lose our ability to participate in the system."
She wants Primavera's seat, and she is convinced that her inclusive Libertarian agenda will resonate with voters tired of politics as usual.
Kim Tavendale immigrated from New Zealand in 1999 and is now a U.S. citizen.
Campaign website: KimForColorado.com
Washington Post article shows Gov. Gary Johnson a guardian of taxpayer money
Well-known political statistician: It's 'plausible' the election could end in a tie


Early childhood development isn't just about creating baby Einsteins. It's important to teach soft skills, such as playing well together, solving problems and being organized. We'll consult experts as we continue to look at the impact of the First 2000 days on a child's life. We'll also learn about the impact of toxic stress on young people, and intervention strategies. And, a Cavs championship parade preview, on The Sound of Ideas.


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Talk:Adi Da
I think it's important that quotes about this interesting and controversial guru reflect experiences of present and former students, who have had direct experience of him and his religious community, as well as the opinions of relatively "famous" people who know him primarily through his writings. To that end, I've restored a critical quote deleted by a user who (on Wikipedia) has in the past deleted material critical of Adi Da. —Jim Butler 2005 June 3
categorizaionEdit
How should we categorize him? Wikipedia put him under "New religious movements". I got an impression "religious leaders" is inappropriate, neither thinkers or philosophers. But I am not sure "movements" is a good grouping for people. --Aphaia 6 July 2005 06:15 (UTC)
I don't see why "leader" is bad. If people follow him, he's a leader -- no matter how cranky we happen to think his religion is. ~ MosheZadka (Talk) 6 July 2005 06:56 (UTC)
The problem is, not the part of "leader" but "religious". Many new age thought people protest to be attributed "religious" or gives no comment if such attribution is adequate. And I have no idea if he is thought as such. For example, English Wikipedia doesn't classify him as "religious leader"... --Aphaia 6 July 2005 07:05 (UTC)
Our intro text currently says "the founder of the new religious movement known as Adidam". So we already blame "Adidam" for being a religion, might as well be consistent about it. ~ MosheZadka (Talk) 6 July 2005 07:23 (UTC)
I concur. --Aphaia 6 July 2005 07:37 (UTC)
Verifiability, etc (edits on April 1st 2006)Edit
Added latest verified Wilber quote. Updated links, using most verifiable ones where possible. Qualified sources when likely verifiable (by virtue of having been published in some other verifiable source in the past) but hosted on POV or personal sites; some such sites have online copies of stuff that has been published elsewhere (like the Watts and Wilber book blurbs), and those seem fine. The Regardie quote, however, needs an original source to be verifiable, as may a couple others from the About Adidam site. I am assuming that devotees like Dan Roberts have consented to have their words used there, so that it's in effect a primary source for devotees. But other folks like Regardie need to be verified.
Quotes that has not been published elsewhere may not be verifiable by Wikipedia standards: e.g., the unpublished Wilber letter to Adidam and probably the Broken Yogi (Conrad Goehausen) Lightmind posts. Mr Goehausen will undoubtedly vouch that the quoted words are his own, but they're not notable enough and it's not worth the trouble: he has since published his own blog and makes the same points there, so that materiial can be used instead (I'll do this later). The Wilber Adi Da letter I don't have a problem with in "truth" terms, but in verifiability terms, I'm not sure. Has Wilber vouched for it?
Adi Da is a guru with a relatively small following (1-2K current, ca. 10K ex, estimated) and it's understandable that "experts", i.e. people who have been directly involved, would tend to self-publish their stuff. So the self-published sites and blogs are cool as primary sources. I trimmed the Jim Chamberlain quote; it's offlinked anyway.
I am tagging the Alex Grey and Godfree Roberts quotes which do need citations, and will delete if no one sources them. It's fine if Godfree Roberts chooses to self-publish his thoughts somewhere, and his quote is valuable, but Wikipedia itself cannot function a primary source for self-publication of such things. thanks, Jim Butler 07:03, 2 April 2006 (UTC)
P.S. I'm having difficulty, obviously, tagging the quotes that need citations. Ther tags that work on Wikipedia don't work here, such as {{fact}}, which just leads to en empty Template page on Wikiquote instead of the functioning page on Wikipedia. Maybe there is a Wikimedia-specific template page; need to find out. Jim Butler 19:06, 2 April 2006 (UTC)
Quotes page clean upEdit
The wikiquote page on Adi Da has been used as a anti- pov propaganda page up to this point and is a mess for the reader or any one seeking quotes by Adi Da Samraj , cannot see any need to have quotes about Adi Da (rather than by Adi Da) here at all (from any source --if nessacery these could be external links at bottom of page --) after looking at many Bio Article quote pages , this approach seems out of step with how this page is generally used ( wikiquotes ) so will edit out the current format --Scribe5 21:41, 26 April 2006 (UTC)
Thank you for your work on the page but it's not appropriate to delete quotes by others about Adi Da as these exist on other wikiquote pages. I restored the section and will fill out over time Carry18 01:10, 30 April 2006 (UTC)
In my opinion they really have no useful place here only personal propaganda material ( pro or anti) , if you choose to use them , one liners and very few should suffice , otherwise will edit them to a useful length, best if you do this to avoid conflict --Scribe5 01:49, 30 April 2006 (UTC)
I will be happy to comply with your suggestions if you can show a basis for them. Otherwise we should follow guidelines like this one. AFAIK there is no special limit on quote length (if there is, please practice what you preach!). Quotes should be notable. Miller is a notable critic of Adi Da. Carry18 03:01, 30 April 2006 (UTC)
Miller is a notable critic of Adi Da but Miller is not noteworthy, is he a famous person of any description? other than an ad homien critic of Adi Da , the fact that he is "the champion" used by other non noteworthy critics does not add up to being famous or of note , you may as well say 'joe bloggs" got a letter in to the editor in 1960 , therefore he is some one of note , also check your Cromwell page only famous people there . To bring quotes in merely to defame someone is just not right, no matter what specious arguemets you use. My point is made clearly will tag this page and get outside views if you persist
I think critical quotes are allowed under NPOV. Miller's statements refer to sworn statements and allegations covered in major media so I do not think they can be considered defamation. So please feel free to tag and request assistance, thank you Carry18 03:41, 30 April 2006 (UTC)
Also realize that the problems that Wilber refers to are exactly the controversial events of 1985 which involved Miller and others. Da is himself not that notable. But when speaking of Da, the 1985 critics are notable. That would be my interpretation. Carry18 03:45, 30 April 2006 (UTC)
Retrieved from "https://en.wikiquote.org/w/index.php?title=Talk:Adi_Da&oldid=212340"
Return to "Adi Da" page.


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The Umbrella Academy fans think series just revealed a major change for season three
Louis Chilton
March 16, 2022, 11:59 AM ·1 min read
Fans of The Umbrella Academy think that Netflix has just hinted at a major change of direction for the series going forward.
The change in question concerns the show's age rating. The popular Netflix sci-fi show, which stars Elliot Page, Robert Sheehan and Tom Hopper, had been classified as a TV-14 rating in the US for its first two seasons.
A TV-14 rating usually equates to a 12 rating by the UK guidance board, the BBFC.
However, ahead of the third season's release, this has now been changed to TV-MA rating (not suitable for viewers under 17) – suggesting that the graphic violence, bad language and sexual content in the new episodes have been turned up a gear.
Fans shared their reactions to the news on Twitter.
"About time!" wrote one viewer. "This deserves a TV-MA story like Doom Patrol."
"They bringing the violence for season 3??????" speculated another.
"Ok hello im literally so excited," wrote someone else, while another fan exclaimed: "OKAYYYY IM EVEN MORE EXCITED NOW."
One fan simply put: "THANK. GOD."
The Umbrella Academy season three arrives on Netflix on Wednesday 22 June. The previous two seasons can be watched on the streaming service now.
France, Australia to supply Ukraine with artillery shells
Memphis weather: Live updates on ice storm, power outages, closures and more


Join Oldfather Group Agent Pamela Foster this Saturday, August 19, from 11 a.m. to 2 p.m., as we showcase this welcoming home in the Indian River Bay community of Oak Orchard.
Located at 33188 Pearl Street, this inviting coastal home features four bedrooms and two bathrooms and an eat-in kitchen with views of the bay.
The property also boasts an open floor plan above a two-car garage and just looks the part of a coastal Delaware beach home. There's also a deck and a patio that both offer views of the water and the community.
This coastal home will be open on Saturday, August 19, from 11 a.m. to 2 p.m., so be sure to stop by and have a look.
And if you'd like to see this or any of the other great homes available in southern Delaware, please give us a ring at 302-260-2000. We'd love to welcome you into the Oldfather Group family.


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Showing 91-100 of 169
The Masquerade looks like an ordinary cocktail, but once you peel away the mask you realize is an incredible drink packed with flavor and aroma. Made form Gran Gala Triple Orange liqueur, Pisco, lime juice, orange bitters, egg white and cardamom, and served in a chilled rocks glass.
Merry Widow Fizz
Celebrate the passing of a spouse with a toast. The Merry Widow Fizz drink recipe is made from Dubonnet Rouge, orange juice, lemon juice, egg white and club soda, and serve dover ice in a Collins glass.
Mini Sour
The Mini Sour is a pair of Saint Patrick's Day shots made from Jameson Irish whiskey, lemon juice, simple syrup, bitters and egg white, and served in chilled shot glasses.
Montezuma was an emperor of the Aztec empire. The Montezuma Cocktail is a full-bodied Cinco de Mayo drink recipe. Made from gold tequila, madeira wine and coddled egg, and served in a chilled cocktail glass.
Morning Glory Fizz
The Morning Glory Fizz drink recipe is made from scotch, Pernod absinthe, lemon juice, sugar, egg white, bitters and club soda, and served over ice in a highball glass.
Mountain Cocktail
The Mountain cocktail recipe is an earthy drink made from blended whiskey, dry vermouth, sweet vermouth, lemon juice and egg white, and served shaken in a chilled cocktail glass.
My Flip
In general, any of the 'flip' cocktails use some part of an egg with an aged spirit. The My Flip cocktail recipe is made from cognac, raspberry liqueur, Wenneker Cinnamon Red liqueur and egg yolk, and served in a chilled cocktail glass garnished with freshly ground nutmeg.
New Orleans Gin Fizz
The New Orleans Gin Fizz drink recipe is made from gin, lime juice, lemon juice, half-and-half, simple syrup, egg white and club soda, and served over ice in a rocks glass.
New Year's Fizz
The New Year's Fizz is likely one of the best New Year's Eve drink recipes. Made from Wenneker Mandarin orange liqueur, gin, prosecco, lime juice, ginger juice, ginger syrup, egg white, basil and pineapple, and served over ice in a highball glass garnished with pineapple and basil.
The Nightingale cocktail recipe is made from banana liqueur, triple sec, light cream and egg white, and served in a chilled sherry glass.


A Beefsteak Tomato is a large fleshy variety that we vine ripen in a Greenhouse to produce the fullest flavor and quality of a great eating tomato. This large tomato will have the distinctive bright green stem (calyx) still attached. Choose them at a red or pink color, with no bruising or excessive softness. Store on the kitchen counter at room temperature and consume at a bright red color. Avoid putting your tomatoes in cold refrigerated temperatures as this will lower the flavor.
Temperature: 55-70 F, 12.8-21 C. No Mist.
Typical Shelf life: 7-14 days.
Highly sensitive to freezing and chilling injury.
Humidity higher than 85-88% will encourage decay.
Continues to ripen after harvest.
Good quality tomatoes should have bright, shiny skins and firm flesh.


text:
Digital game downloads are bad for the surroundings than Blu-ray discs
Do you think that the world would be a less contaminated place if all and sundry switched to digital downloads, decreasing the number of physical objects — and factories to manufacture them — in the world. A recent study suggests that despite the fact that eradicating physical objects lowers the potential for future for junk and factory pollution, going digital isn't the best response to clean-up up the planet — in reality, the carbon footprint of downloading games is in fact higher than purchasing a physical copy.
The study, published in the Journal of Industrial Ecology, focuses on the carbon footprint of games distribution — specifically, an 8.8GB PlayStation 3 game — using the metrics of a typical UK production and distribution cycle. When calculating carbon emanation, there are more issues that go into both the physical retail and digital cycles than it may appear. Aside from the palpable metric of factories producing the discs and packaging, other issues — such as the vehicles used to transport the products, and the output produced when ultimately disposing of the items — adds to the overall trail. The similar thing pertains to digitally distributed games, for instance, if televisions and monitors are powered on during a long download — something that doesn't apply to acquire a retail disc.
Startlingly, it turned out that the carbon emanation of a retail disc — including the production, vehicle delivery, and so on — was in fact lesser than that of a downloaded game.
Carbon emissions for the 8.8GB PS3 disc were anticipated to be around 20.82 kilograms, but a corresponding digital download reached 27.53 kilograms. A variety of factors add to the increased carbon footprint of a downloaded game, such as the electricity consumed by the downloader, the power used by data centers to send the content, and even the power which is used to produce the digital file in the initially. The study does confess though, that it had to make quite a few hypotheses for one rationale or another. As the study states, for example, the type of software used, the reason of a download, and the resources accessible to any given company are not entirely accessible on an individual basis. Nevertheless, there are available reasonable averages that can be used.
The study does look to the future, but exclaims that it's too premature to tell to see if carbon emanation of digital games will stay alike or reduce. In theory, internet competence will boost in the future, thus lessening the carbon footprint of a downloaded title. Nevertheless, games usually increase in file size, and it's hard to distinguish whether or not internet speeds will swell enough in turn to lower download times in spite of the larger file sizes. Other factors that are presently becoming extensive, for example, remote downloading while you're travelling home from work or being capable to use your console while a game downloads in the background, will also play significant roles in the game industry's carbon footprint.


It's an unfortunate fact that finances and insurance coverage often limit people's ability to choose the treatment option they would prefer. We established the CLEAR Family Fund specifically to help people overcome these barriers to receiving treatment. 100% of donations to the CLEAR Family Fund go towards empowering a deserving patient to obtain the care they need. If you or someone you love has been helped by chiropractic or the CLEAR methods, please consider donating to help someone suffering from scoliosis experience a similar life-changing experience. Your tax-deductible gift will relieve some of the stress and financial burden that can come with a diagnosis of scoliosis, and the stories of the patients who benefit from your donation will be shared in CLEAR's blog so that you can see the difference you made in their life.
The journey to living a full life with scoliosis relies on a strong network of support, including family, friends and people like you. And the CLEAR Scoliosis Institute is ready to build on that support by offering highly customized care with proven results.
Our first milestone is within reach! Can you help us close the gap? The fund only has $150 left to go before we can offer treatment to a scoliosis patient in need.
Will you help a family today?
Join us in offering CLEAR treatment to scoliosis patients looking for the opportunity to live a better life. You can also support the CLEAR Family Fund by making a purchase in our new online store.
I would like to recieve emails from CLEAR.


Most eukaryotes contain many tandem repeats of ribosomal RNA genes of which only a subset is transcribed at any given time. Current biochemical methods allow for the determination of the fraction of transcribing repeats (ON) versus non‐transcribing repeats (OFF) but do not provide any dynamical information and obscure any transcription activity at the single‐cell level. Here, we use a fluorescence in situ hybridization (FISH) technique that allows the detection of single‐RNA molecules in individual yeast cells. We use this method complemented with theoretical modeling to determine the rate of switching from OFF to ON (activation rate) and the average number of RNA molecules produced during each transcriptional burst (burst size). We explore how these two variables change in mutants and different growth conditions, and show that this method resolves changes in these two variables even when the average rDNA expression is unaltered. These phenotypic changes could not have been detected by traditional biochemical assays.
Fluorescence in‐situ hybridization (FISH) technique allows the detection of single RNA molecules in individual yeast cells. We use this method complemented with theoretical modeling to determine the rate of switching from OFF to ON (activation rate) and the average number of RNA molecules produced during each transcriptional burst (burst size).
Switching of the rDNA repeats between the inactive non‐transcribing state and the active transcribing state occurs rapidly. On average, it takes an inactive repeat 8 2 min to transit to the active state.
As cell density increases, mean rRNA transcriptional activity decreases. This is due to the decreases in burst size. The activation rate remains constant.
In the rpd3 strain, activation rate doubles and the burst size is half that of wild‐type cells in log‐phase. This effect of Rpd3 on activation rate is independent of cell density. As the cell density increases, the burst size of the rpd3 strain becomes similar to that of wild‐type cells.
Regulation of rDNA synthesis is important for cell growth control (Warner, 1999; Grummt, 2003). Yeast cells carry about 150 copies of tandemly repeated rRNA genes. The regulation of rDNA transcription occurs by controlling both the fraction of actively transcribing repeats and the transcription rate of the active genes (Sandmeier et al, 2002; French et al, 2003). During rapid growth, only about half of these genes are active (ON) and being transcribed (Conconi et al, 1989; Dammann et al, 1995). The remaining repeats are inactive (OFF) and are not transcribed. Two distinct chromatin states correlating with the difference in transcriptional activity has been observed (Dammann et al, 1993). It has been shown that OFF repeats are arranged into regular nucleosomal arrays that prevent access of the Pol I transcription machinery, whereas ON repeats have an 'open conformation'. More recently, it has been shown that active repeats exist in a dynamic chromatin structure of unphased nucleosomes, associated with chromatin‐remodeling enzymes that may act to facilitate efficient rRNA synthesis (Jones et al, 2007). Current biochemical techniques like in vivo psoralen (Dammann et al, 1993) and electron microscopy (EM) Miller spread (Miller and Beatty, 1969) assays are frequently used in the study of rDNA regulation. Psoralen selectively cross‐links to ON repeats and slows their migration during electrophoresis, allowing the determination of the fraction of ON repeats. rDNA regulation studied using EM by the Miller spreading method allows visualization of active and inactive rDNA genes. These methods have greatly enhanced our understanding of rDNA regulation but they are tedious, require lysis of cells and do not give any information on the kinetics of regulation. Furthermore, these studies have primarily focused on population statistics. Here, we use a sensitive fluorescence in situ hybridization (FISH) method that allows visualization of single‐RNA molecules in individual cells. We obtain distributions of rDNA transcriptional activity in single cells and use these to gain insight into the kinetics of regulation and to investigate how the kinetics changes as the cell adapts to different conditions.
To monitor rDNA transcription in single‐yeast cells, a synthetic construct containing 32 tandem copies of a 43–base‐pair probe‐binding sequence was inserted downstream of the 35S promoter into one of the repeats in the rDNA array (Figure 1A). These cells were then fixed and hybridized with a single‐stranded oligodeoxyribonucleotide probe, labeled with five tetramethylrhodamine (TMR) fluorescent molecules, which was complementary to the tandem repeated sequences. The binding of many fluorophores to each RNA molecule allows its detection as a diffraction‐limited spot in a conventional wide‐field fluorescence microscope (Femino et al, 1998; Raj et al, 2006). We observed cells with no or few RNA molecules and also cells with many RNAs and bright transcription sites (Figure 1B, top‐right panel). These correspond to cells with silenced and active tagged repeats, respectively. The bright transcription sites observed are consistent with EM Miller spreads that show many polymerases transcribing from an active gene producing many RNAs.
Genetic manipulation, methods and modeling. (A) A synthetic construct is inserted behind the 35S promoter in one of the repeats in the array to report the transcriptional activity of that repeat. Probes labeled with TMR fluorphores are designed to bind to the RNA transcribed from this repetitive synthetic construct. (B) (Top‐left) Bright‐field image of the fixed yeast cells. (Bottom‐left) A cell segmentation program is used to identify individual cells. (Top‐right) Fluorescence images of cells that had been fixed and hybridized with fluorescent probes. Some cells have bright transcription sites and the individual RNA cannot be resolved as single dots. Other cells have fewer RNA that can be resolved. (Refer to Supplementary Figure S8 for more information.) (Bottom‐right) Image obtained after processing the TMR image on top‐right. First, a program is used to identify cells with bright transcription sites whose RNAs cannot be resolved. These cells are labeled red. The remaining cells are outlined in green and an image processing program is used to count the number of RNAs in them. The individual RNAs are shown in green. (C) Gene expression is modeled by having an ON state of high transcription and an OFF state of no transcription. Gene activation is modeled by the parameter α, which is the transition rate from the OFF state to the ON state. Gene deactivation is modeled by the parameter γ, which is the transition rate from the ON state to the OFF state. μ is the rate of transcription when the gene is in the ON state. RNA is degraded at a rate δ. (D) Simulation of the steady‐state distribution obtained for different switching rates.
For cells with few RNAs, the RNAs are spatially resolved as individual spots. However, for cells with RNA counts above 40 or cells with bright transcription sites, the RNA molecules are too close to each other to be resolved. Using the cells with low RNA counts, we obtained a linear relationship between the total fluorescence and number of RNAs (Supplementary Figure S1) and estimated the number of RNA molecules in bright cells by linear extrapolation from their total fluorescence. This process is automated by a custom‐designed MATLAB image processing code. The RNA distribution obtained for wild‐type cells is shown in Figure 2A. To determine whether genome position within the rDNA locus has an important function in regulating transcription activity, five different colonies obtained after plasmid integration were examined. As the construct integrated randomly along the array, these colonies have the construct integrated at different positions. The distributions of the five different integrants were similar (Supplementary Figure S2), indicating that position effect does not affect transcription significantly. Colony 1 was selected for further studies.
Transcriptional changes during diauxic shift. During diauxic shift, mean RNA production decreases. This decrease is due to a decrease in burst size. The gene activation rate remains constant. (A) RNA expression profiles of a single‐rDNA repeat at t=0, 90, 120, 180 min after the culture reaches OD600=0.4 were determined using single‐molecule FISH. Red lines are best fit from the burst model. Binsize is 20. (B–D) Mean RNA, normalized activation rates and burst sizes of the rDNA repeat at the different times. Error bars are standard errors from three independent experiments.
The experimentally determined distributions enable us to test dynamical models of transcription regulation and to determine their quantitative parameters. We assume that the burst model (Raj et al, 2006) holds for rDNA transcription. Each repeat can be either in the active state of high transcription or in the inactive rate of no transcription. This model is supported by Miller spread (French et al, 2003) data that shows that each rDNA gene is either silent or transcribing many RNA. This model is characterized by four parameters (Figure 1C). The activation rate α is the rate of switching from the inactive state to the active state. The inactivation rate γ is the rate of switching from the active state back to the inactive state and transcription rate μ refers to the rate of RNA synthesis in the active state. δ is the first‐order rate constant of RNA decay. The three normalized rates, α/δ, γ/δ and μ/δ, characterize the steady‐state distributions. Figure 1D displays calculated distributions obtained at different switching rates.
The model depicted in Figure 1C can be solved analytically (Raj et al, 2006) and was fit to the experimental steady‐state distribution. Because of the long tail of the distributions, the normalized deactivation rate γ/δ and the normalized transcription rate μ/δ cannot be obtained independently. However, the normalized activation rate α/δ and the burst size μ/γ, which is the ratio of transcription rate over gene inactivation rate can be obtained accurately. The burst size μ/γ corresponds to the average number of RNA molecules that are transcribed in each gene activation event. We obtained an average of α/δ=(0.42±0.07) and μ/γ=(190±20). We determined the decay rate δ of the RNA directly in an independent experiment by using thiolutin, an inhibitor of RNA synthesis, and measuring the amount of RNA left at various times after transcriptional inhibition (Supplementary Figure S4). From these measurements, we obtained δ=(0.28±0.03) per min. This yields α=(0.12±0.02) per min, corresponding to an average time of (8±2) min for an inactive repeat to transit to the active state. Earlier, psoralens cross‐linking results have shown that at log‐phase in minimal medium, about 40% of the repeats are active (Dammann et al, 1995). Hence, an active repeat transcribes RNA for about 5 min before transiting back to the inactive state.
Next, we explored how rDNA synthesis responds to perturbations in external conditions. It is known that rRNA synthesis decreases in response to nutrient downshift (Sandmeier et al, 2002). To determine whether the decrease is due to a decrease in normalized activation rate α/δ or burst size μ/γ, a culture starting at OD600=0.4 was monitored for 180 min. We measured the RNA distribution of cells at t=0, 60, 90, 120, 150 and 180 min and fitted the distributions to the steady‐state solution of the burst model. Because of fast decay rate of the RNA as compared with the rate of change of the other parameters, the distribution of RNA closely reflects that of the distribution at steady state. Hence, fitting the RNA distributions to the steady‐state solution is a good approximation (see Materials and methods; Supplementary Figure S6). As expected, the mean RNA counts decreases as the cell density increases (Figure 2A and B). Using the model, we infer that the normalized activation rate α/δ remains fairly constant, whereas the burst size, μ/γ decreases (Figure 2C and D; Supplementary Table I). We measured the RNA half‐life to be (3.4±0.8) min for the cells at t=180 min (Supplementary Figure S4). This decay rate is similar to the half‐life (3.6±0.5) min measured at early log‐phase, suggesting that activation rate α is constant throughout the 180 min of the experiment. Hence, the decrease in rRNA synthesis is due to the decrease in burst size. Earlier work has shown that the fraction of open repeats decreases during diauxic shift and that Rpd3, a histone deacetylase, is responsible for this decrease (Sandmeier et al, 2002). As our measurements show that the activation rate α remains constant, we deduced that Rpd3 decreases the fraction of open repeats by affecting the deactivation rate γ.
To further explore the role of Rpd3 in controlling rDNA transcription, we examined the effect of deleting Rpd3. Despite Rpd3's role in diauxic shift, little is known about its effect during log‐phase. Indeed, it seems like Rpd3 does not affect rDNA transcription in the log‐phase as both the fraction of open repeats and average rRNA concentration in wild‐type and rpd3Δ strains are similar (Sandmeier et al, 2002). To determine its effect during the log‐phase, rpd3Δ strains were grown to OD600=0.4 and fixed for hybridization. Consistent with earlier data, we observed that the mean of the rpd3Δ strain is similar to wild type (Figure 3B). However, the RNA distribution is different (Figure 3A). After analyzing the experimental distribution with the burst model, we find that the activation rate α in the rpd3Δ cells is twice that of wild type, and its burst size μ/γ is reduced by half (Figure 3C and D). The change in the activation rate α and burst size μ/γ in the rpd3Δ cells could not have been revealed by traditional biochemical methods, which measure population averages. We further measured the RNA distributions for the rpd3Δ strain at t=90, 150 and 180 min after it reached OD600=0.4. Unlike the increase in activation rate α, which is constant in the rpd3Δ strain at the different time points, the burst size μ/γ becomes more similar to that of wild type as cell density increases (Supplementary Figure S9A; Supplementary Table II).
Effect of Rpd3 deletion. Although the rpd3Δ strain has similar mean RNA expression as wild type, its activation rate is double and its burst size is half that of wild type. (A) RNA expression profiles of a single‐rDNA repeat in the rpd3Δ strain (red) and wild type (blue) at OD=0.4. Best fits from the model are shown in dash lines. Binsize is 20. (B–D) Mean RNA, burst sizes and normalized activation rates of the wild‐type and rpd3Δ strains. Error bars are standard errors from two independent experiments.
Here, we used transcript counting to study the kinetics of rDNA transcriptional regulation. Our measured switching rate of the rDNA repeat is faster than reported switching rates of markers inserted into other regions that show gene silencing like the telomeres (Gottschling et al, 1990) and the mating locus (Xu et al, 2006). Our results suggest that each repeat spends about 5 min in the active state producing about 190 RNA molecules before switching off. This yields a Pol I reinitation interval of 300/190=1.6 s, which is comparable to the 1.2–2.2 s estimate obtained using EM Miller spreading (French et al, 2003). It has been suggested that active chromatin structure cannot be directly inherited as replication of transcriptionally active rDNA gene results in two newly replicated inactive coding regions packaged into nucleosomes (Lucchini and Sogo, 1995). Here, we suggest that not only the subset of active chromatin is not inherited but the set of active repeats is changing within one cell cycle. We ran a simulation to obtain the probability density for the amount of time that each repeat is open per cell cycle using different switching rates (Supplementary Figure S7). With the inferred switching rates, almost all the repeats will be active in each cell cycle. It has been shown that repair of active genes occurs faster than silenced genes (Conconi et al, 2002). Hence, one advantage of fast switching may be to allow damaged repeats to be repaired before it is inherited.
Population measurements performed on rpd3Δ and wild‐type strains showed that the two strains have similar fractions of active genes and mean transcriptional activity during log‐phase. However, our single‐cell measurement shows that the activation rate doubles and the burst size is reduced by half in the rpd3Δ strain. As the fraction of open repeats in the rpd3Δ strain is reported to be similar to that of wild type (Sandmeier et al, 2002), we suspect that the deactivation rate would have also doubled. This would explain the reduction in burst size if transcription rate is unaffected by Rpd3. This suggests that Rpd3 functions to decrease both the activation and deactivation rates in log‐phase. There are evidences showing that Rpd3 can function both as a transcriptional activator and deactivator (Deckert and Struhl, 2002; Sertil et al, 2007). If we assume that the transcription rate is unaffected by Rpd3, we can normalize the burst size in the wild‐type strain to that of the rpd3Δ to obtain the ratio of their deactivation rates γrpd3Δ/γWT at different time points. This ratio becomes closer to 1 at later time points. This suggests that unlike the effect of Rpd3 on the activation rate α, which is independent of cell density, the effect of Rpd3 on the deactivation rate γ decreases with cell density. One hypothesis for how Rpd3 can regulate activation and deactivation rates independently is that Rpd3 uses different mechanisms to control them. For example, Rpd3 may decrease the activation rate α by deacetylating the H3 or H4 subunits of UAF, a transcription factor for Polymerase I recruitment (Keys et al, 1996) and increase the deactivation rate γ by affecting the nucleosomes surrounding the rDNA promoter. This suggests that in wild‐type cells, Rpd3 does not inactivate ON repeats as cell density increases. Rather, Rpd3 actively acts to repress the deactivation rate γ at low cell density and stops repressing at high cell density, thus enabling the deactivation rate γ to increase (Supplementary Figure S9B). Future work is needed to elucidate the detailed molecular mechanism through which Rpd3 affects these rates.
The 'R's represent locations where an amino‐dT was introduced in the place of a regular dT. The oligonucleotides were synthesized on a controlled pore glass column (Glen Research) that introduced an additional amine group at the 3 end of the oligonucleotide. The probes' amine groups were then coupled to the TMR (Molecular Probes, Eugene, OR). The probes were purified on an HPLC column to isolate oligonucleotides displaying the highest degree of coupling of the fluorophore to the amine groups.
A plasmid with the promoter of 35S driving the synthetic repeat was linearized with HindIII, which cuts the plasmid within the promoter region. This plasmid was then integrated into haploid W303a by plasmid integration. Transformations were performed with different concentration of the plasmid using URA3 as the selectable marker. Colonies on the plate with the lowest plasmid concentration yielding successful transformants were selected.
To ensure that the RNA counts we obtained do not include partial degraded RNAs, a Northern blot was performed using the FISH probe sequence. As observed in Supplementary Figure S3A, the Northern blot yields a single band, showing that the transcripts detected by FISH are full length. We noticed that the length of the transcripts is about 3 kb, which is longer than the 2 kb reporter sequence in the plasmid. This is not surprising as Polymerase I transcriptional read‐through of a polymerase II termination site has been reported earlier (Lo et al, 1998). The sequence downstream of the reporter sequence is part of the URA3 sequence in the plasmid downstream of the termination site.
To check that the reporter is integrated within the rDNA repeats, we performed a PCR using a primer binding to the AMP sequence on the plasmid and the other primer binding to a region of the 35S sequence. This PCR reaction detects a band of 2 kb, showing that the reporter is integrated within the rDNA repeats (Supplementary Figure S3B).
For confirming that a single plasmid has been integrated, quantitative PCR was performed using primers against URA3 and ACT1. W303a has a single copy of URA3 gene. Integration of the plasmid leads to additional copies of URA3 being integrated into the genome of W303a. This will lead to a reduction in the number of cycles needed to generate similar amount of the URA3 PCR product. We compared this cycle difference in the strains before and after plasmid integration. A change in cycle number of −0.9±0.3 is obtained after plasmid integration, showing that one copy of the plasmid has been inserted (Supplementary Figure S3C). ACT1 was used as a loading control.
To generate the rpd3Δ strain, primers with 50 bp homology to the beginning and end of the coding sequence of Rpd3 were used to PCR out a sequence containing the LEU2‐coding sequence. This PCR product was then transformed into the wild‐type strain and plated on plates lacking leucine. Colonies selected are those with the LEU2 replacing the RPD3 gene and were confirmed by PCR.
Yeast cells (haploid W303a) were grown in CSM‐URA minimal media at 30°C to an OD600 of 0.4 (for log‐phase experiments). Cells were fixed by adding formaldehyde to a final concentration of 4% (v/v) for 45 min at room temperature (23±2°C). The cell wall was digested with lyticase at 30°C for 20 min. After that, cells were stored in ethanol at 4°C. Before hybridization, cells were rehydrated in 30% (v/v) formamide, 2 × SSC for 10 min. Cells were hybridized in 30 μl of hybridization solution containing labeled DNA probes in 25% (v/v) formamide, 2 × SSC, 1 mg/ml BSA, 10 mm VRC, 0.5 mg/ml Escherichia coli tRNA and 0.1 g/ml dextran sulfate overnight at room temperature. Before imaging, cells were washed twice in 30% (v/v) formamide, 2 × SSC for 30 min. DAPI was added to the first washing step. The cells were then added to multichambered coverglass (Lab‐Tek, Nalge Nunc, Rochester, NY) coated with concanavalain A (0.1 mg/ml, Sigma) for 10 min. Cells that did not stick to the coverglass were washed off.
Images were taken with a Nikon TE2000 inverted fluorescence microscope equipped with a 100 × oil‐immersion objective and a Princeton Instruments camera using MetaMorph software (Molecular Devices, Downington, PA). The TMR filter was obtained from Chroma (Rockingham, VT). Exposure time for TMR was 1 s. Stacks of images were taken automatically with 0.2 microns between the z‐slices.
To segment the yeast cells, a watershed algorithm was used. Cell boundaries were obtained by running an edge detection algorithm on the bright‐field image of the cells. After that, individual cell nuclei were identified in the DAPI images by (1) selecting pixels above a certain threshold, (2) connecting these pixels that are close to each other and (3) rejecting connected regions that are larger or smaller than certain thresholds. These nuclei serve as markers for running a marker‐controlled watershed algorithm over the bright‐field image. Next, a program was run to count the number of RNA molecules in each cell. The general procedure was to run a median filter followed by a Laplacian filter on each optical slice taken. This enhanced particulate signals. A threshold was then selected to pick‐up individual spots in each plane. Next, the program counted the total number of isolated signals (i.e. connected components) in three dimensions. The particle count was robust for a range of threshold chosen. This particle count was accurate for cells with low number of RNA. For cells with RNA counts larger than 40 molecules, the RNAs were too close and could not be resolved as individual dots. To identify these cells, a program was run to determine cells with large connected regions of high fluorescence intensity. These regions consisted of the many individual RNAs that could not be resolved as individual dots. Next, the total cell fluorescence was obtained by summing up the fluorescence in each plane. By fitting the total cell fluorescence and the number of RNAs in cells with RNA counts smaller than 40 molecules, a linear relationship was obtained between cell fluorescence and RNA count. This relation was extrapolated to determine the number of RNAs in cells with high total cell fluorescence.
To determine the parameters of our model, we fit our experimental data to the steady‐state solution to the master equation of the two‐state model using a maximum likelihood method. We found that the likelihood value is similar for similar ratios of μ/β. This indicates that there was insufficient experimental data to resolve the two parameters. The error bars refer to 95% confidence interval. For more information on the details of the two‐state models, please refer to Raj et al (2006).
A measure of 8 μg/ml of Thiolutin, an inhibitor of RNA synthesis, was added to the culture at time t=0 min. Thiolutin is a metabolite produced by Streptomyces luteroeticuli that inhibits all yeast RNA polymerases, mainly at the level of transcriptional initation. Cells were collected at t=0, 1, 2, 3, 4, 5, 10, 15, 20 and 40 min after thiolutin addition. The mean RNA count at each time point was measured using FISH and fitted to the equation m=m0e−dt+b, where m(t) is the mean RNA at different times after thiolutin addition. m0 + b is the initial amount of RNA before thiolutin addition. d is the rate constant for RNA decay and b is the amount of RNA transcription after thiolutin addition. The best fit was determined by the least square method with m0, d and b as free parameters. To determine the error in the fit, we randomized the RNA count at each time points by drawing random number from normal distributions with the measured mRNA and standard derivation for that time point. This set of RNA obtained from the randomization was fitted to the decay equation using the least square fit. This process was repeated 100 times and the standard derivation obtained from these 100 sets of parameters was used as the estimate of the error in the parameters.
We compared the decay rate measured using thiolutin inhibition to that measured using temperature inhibition in a cbf5‐1 temperature sensitive strain (Cadwell et al, 1997). In the cbf5‐1 strain, rDNA synthesis is inhibited at the non‐permissive temperature of 38°C. To measure decay rate by thermal inactivation, the cbf5‐1 strain was first grown to OD600=0.6 at 27°C, spun down and resuspended in an equal amount of medium preheated to 38°C. The cells were then removed and fixed at t=1, 2, 3, 4, 6, 10 and 20 min after transfer.
The half‐lives measured using thermal inactivation and thiolutin inhibition are (1.5±0.2) min and (1.9±0.3) min, respectively (Supplementary Figure S5). The two half‐lives are similar. The slight difference in the decay rate measured may be due to the fact that enzymes for decaying the RNA may be more reactive at 38°C as compared with 27°C. It is also interesting to note that the decay rate of the reporter construct is faster in this strain as compared with our experimental strain. This discrepancy may be due to the differences in strain backgrounds and the fact that our strain is a haploid whereas the cbf5‐1 strain is a diploid.
For the diauxic shift experiments, cells were taken out of a culture at t=0, 60, 90, 120, 150 and 180 min after the culture reached OD600=0.4. The cells were then fixed and hybridized for imaging.
During diauxic shift, the mean mRNA decreases rapidly. To check whether the steady‐state assumption is valid, we ran a simulation modeling the decrease in RNA expression and fitting the RNA distributions obtained to the steady‐state equation.
To simulate the RNA decrease, we estimated the activation rates and burst sizes from the steady‐state fits obtained for the experimental RNA distributions at t=0, 60, 90, 120, 150 and 180 min during the diauxic shift. As the activation rate remains relatively constant, we used a constant activation rate for the simulation. For the burst size, we assumed that it is piece‐wise linear. Using these estimates for the activation rate and burst size, we were able to construct the time‐dependent master equation matrix and run a discrete time Markov process to model the RNA distribution for the 180 min duration of the experiment. To fit the simulated RNA distribution at t=60, 90, 120, 150 and 180 min, 1000 cells were randomly drawn with RNA numbers reflecting the RNA distribution at that time point. The maximum likelihood method was then used to obtain the best fits for the distribution. This was repeated three times to obtain the average activation rate and burst size.
As seen in Supplementary Figure S6, the activation rates and burst sizes obtained from the steady‐state fits are similar to the actual activation rates and burst sizes at each of the time points. This shows that the steady‐state approximation can be used to fit the distributions.
We thank J Gore, G Neuert, B Pando, LF Cheow, M O'Kelly and the van Oudenaarden laboratory for discussions. We are grateful to A Raj for providing the TMR probes. RZ Tan was supported by A*STAR, Singapore. This work was supported by National Science Foundation grant NSF‐0548484 and US National Institutes of Health grant R01‐GM068957.


* The ingredient synergy of Tetrase, oleanolic acid, and rosemary provide efficacy at a low dose for a maximum cost-effectiveness.
* Developed through the ExpresSyn Process, which combines cell proteomic research, safety evaluations, human ex vivo research, and clinical testing for maximum safety and effectiveness.
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Home F1 Teams not blaming Mercedes for recent dominance in F1
Teams not blaming Mercedes for recent dominance in F1
Copyright: Red Bull Content Pool
Darshan Chokhani
Selected teams along with Honda don't blame Mercedes for their top work and dominance in the hybrid era of F1.
In the F1 Beyond The Grid podcast, Ross Brawn revealed one of the secrets for Mercedes to be so ahead in the game. He talked about the manufacturer asking questions and seeking clarifications on certain items with nearly six months gap to their rivals.
It meant, Mercedes already had a commanding start over their then competitors Ferrari and Renault, while despite a one year late start for Honda to prepare themselves as much, it didn't work with the Japanese manufacturer faltering the most.
Since F1 2014 until 2018, Mercedes has taken all the constructors' championship and also the drivers' title – shared between Lewis Hamilton and Nico Rosberg. Ferrari looked like a worthy challenger but so far in 2019, Mercedes has taken a commanding lead.
While we can say Ferrari had their chances but even then the German manufacturer found their way and look set to win both the F1 titles for the sixth time in a row. It has certainly turned off certain section of fans but teams aren't blaming Mercedes for their dominance.
The team from whom, they took the mantle ahead, Red Bull Racing's Christian Horner said: "Everybody has the same set of rules presented to them at the beginning of each year and you've got to take your hat off to Mercedes and Lewis for doing such a great and dominant job.
"I think, for me, the lessons that you learn, particularly over time, is that the more you change things, the more spread you create in the field and the regulations change that we had over the winter didn't help anything.
"In fact it made a dominant team even more dominant so I think the aerodynamic changes, the tyre changes that were made actually didn't do anything to help the sport or the spectacle and I think that's where we need to be very very careful for 2021.
"This is because each team will believe they can get an advantage and ultimately somebody will get it right, somebody will get it wrong but there could be a much bigger spread than there is currently and for me I think the most important thing is once a set of regulations is achieved and found for the future, is consistency.
"The most important thing is leave it alone for a long period of time and then you will get the grid coming together with stability. The worst thing is dicking and changing something every year which isn't fully understood that then changes the dynamics of the car.
"So for me I think you will get absolute convergence with stability and I think that whatever is done for 2021 should be fixed pretty much for a five year period unless there's some fundamental flaw in it."
At the same time, his engine partner, Honda's Toyoharu Tanabe, added: "Mercedes and Lewis are doing a very good job. They are talking about the 2021 regulations and then the PU manufacturers working on 2021 regulations to have a more competitiveness for each person. Not fixed yet but we will see the result soon, I think."
While Red Bull focused on having consistency with the rules instead of constant changes, Renault's Marcin Budkowski pinned his hope on the 2021 rules, especially with regards to the budget cap, which has been an issue for them, even with manufacturer support.
"I don't think you can say something like Mercedes are killing the sport, they are doing a really good job and they are deserving their wins and their championships and all of us are sat here and thinking we should look up to them and do the same job and then we will compete with them.
"I think there are changes coming for 2021 under Liberty's and the FIA's guidance to make the sport more competitive, certainly to make it fairer and more sustainable as well. Whether they will be successful… we think they are on the right path.
"I certainly think the budget cap is probably the biggest impact on the relative competitiveness of the sport so we're very supportive, we're very pro-active in helping them writing the regulations and planning what the sport will be from there.
"Whether they will be sufficiently impactful for all the teams to compete for victories I'm not sure. However, is that what we really want? We want the best working teams to compete for victories. Hopefully there's more than just one or two."
McLaren's Zak Brown more or less agreed with others as well: "You've got to give credit to Mercedes and Lewis for doing an outstanding job. I think we all recognise the sport is not where it needs to be, to have a more level playing and therefore better competition.
"I think 2021 will be a big step forward so I think we can expect a lot more Lewis and Mercedes victories this year, most likely again next year and then hopefully in 2021 with enough rule change that we will start to see the grid get a little bit more mixed up because certainly we need to be putting on a better show for the fans."
Mercedes' customer, Williams' Claire Williams praised Red Bull's work for beating Mercedes in Austria and added on the 2021 rules as well: "I don't think you can take away from Mercedes the extraordinary job that they have done and it's down to the remaining nine teams to do a better job to take the fight to them.
"Someone's got to win, that's the whole point of this sport, that you employ the best people, you get the best resources you can around you in order to create the most competitive car and they have done the best job of that. Someone has to win in this sport.
"We don't obviously want the same team – it's not great for the health of our sport – for the same team to win in each and every race. I think that's why huge congratulations have to go to Red Bull because it's tough to beat Mercedes at the moment and to see another team taking a victory at an event was fantastic, fantastic for the sport.
"But it's about creating great racing, isn't it and making sure that all the regulations collude in order to achieve that and I'm very much hoping obviously that the regulations that are coming on board in 2021, whether that be the technical regulation, the financial regulations all contribute to helping improve the competition that we have, that ensure the sustainability of our sport moving forward into the future."
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As Per Sellers Updated (2018) Stainless Steel Appliances In The Main Kitchen, New Roof And Laminate Floor Done In 2018, Kitchen In The Basement And Updated Lighting Fixtures.
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103 illustrations (96 are b&w and 7 are in color).
UNDERSTANDING LAVA TUBES AND LAVA CAVES takes you inside an active lava flow.
* The two methods of lava tube formation are explained.
* The various internal processes responsible for changing the size and shape of a tube, are revealed.
* You'll learn how air shapes delicate cave features, and changes the color of the rock.
* You'll be amazed by the incredible diversity of formations found in lava caves, and understand how they form.
* And finally, you'll discover that not all lava caves, are lava tubes. Other types of lava caves do exist.
In addition to this, you are given practical information about lava tube caving, and preservation. And there is a Question section which provides further information.
Sections are well illustrated, informative, and interesting, making UNDERSTANDING LAVA TUBES AND LAVA CAVES a must have for anyone wanting to learn about this fascinating environment.
Harry has devoted several thousand hours of the past 21 years exploring lava caves, assisting in cave surveys, and giving lava tube tours. Today, he primarily guides lava tube tours in Kazumura Cave where he teaches visitors about lava tube geology, while stressing preservation.
An impressive presentation of a considerable amount of information regarding the formation of lava tubes; I was quite impressed with the content of this book. It is easy to read and understand the many of aspects of cave formation. They are presented in a format that does not make one feel like you must have a doctorate to "get it". I was fortunate to participate in an on-site tour as well. Harry Shick is very knowledgeable in his field and most capable in relaying the information in everyday language. Taking his tour is just amazing. He is ready and willing to answer all questions presented. I highly recommend this small book.
VERY INFORMATIVE BOOK Review by Kent Henderson.
This very informative book is written by Harry Shick, a guide Kazumura Cave in Hawaii – at 65.5 km reputedly the world's longest known continuous lava cave.
The book deals with Lava Tube Formation; Processes that alter the size and shape of a lava tube; Air – the Second fluid; Formations and Features; and Other Lava Caves. Appendices include Caving in Lava Caves and Preserving the Cave Environment. The book also includes a Glossary, References and an Index. It is profusely illustrated with diagrams and images.
This small book is a good read, and more importantly an excellent text on lava tubes and their formation. I highly recommend it.
After enjoying the Kazumura Cave tour while in Hawaii - a high point of our trip! - I settled down to read Harry Shick's "Understanding Lava Tubes". I thought I had learned a lot while on tour, and was unprepared for the logical thoroughness of the book. Speaking as someone with 30 years professional background in analytic writing, this little book is a masterpiece of clarity. Backed up by a series of clear, simple diagrams and photographs, it makes the mechanics of lava tube formation, and their complex features, flora and fauna easily understood. The author and guide has spent many years studying and investigating such caves first hand, and probably has a unique understanding of them which was obvious in both the book and the tour. Recommended as an essential accessory for anyone who tours Kazumura, but also as a reference for academics and teachers who need to explain lava tubes to students.


Albert Daniel Muir was the youngest son of Daniel and Ellen (née Taylor) Muir. He married Catherine Walker in 1928. His brother John was killed in action in 1917 in France, and another brother Archie served in World War I and was awarded the Military Medal.


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