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No catches, no fine print just unadulterated book loving, with your favourite books saved to your own digital bookshelf.
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Jeanette Winterson OBE is the author of ten novels, including Oranges are not the Only Fruit, The Passion and Sexing the Cherry; a book of short stories, The World and Other Places; a collection of essays, Art Objects as well as many other works, including children's books, screenplays and journalism. Her writing has won the Whitbread Award for Best First Novel, the John Llewellyn Rhys Memorial Prize, the E. M. Forster Award and the Prix d'argent at Cannes Film Festival.
In Brexit Britain, a young transgender doctor called Ry is falling in love - against their better judgement - with Victor Stein, a celebrated professor leading the public debate around AI. Meanwhile, Ron Lord, just divorced and living with Mum again, is set to make his fortune launching a new generation of sex dolls for lonely men everywhere. Across the Atlantic, in Phoenix, Arizona, a cryonics facility houses dozens of bodies of men and women who are medically and legally dead... but waiting to return to life. But the scene is set in 1816, when nineteen-year-old Mary Shelley writes a story about creating a non-biological life-form. 'Beware, for I am fearless and therefore powerful.' What will happen when homo sapiens is no longer the smartest being on the planet? Jeanette Winterson shows us how much closer we are to that future than we realise. Funny and furious, bold and clear-sighted, Frankissstein is a love story about life itself.
'Behind every great woman is a man who tried to stop her.' A century on from the first votes for women in the UK, award-winning author Jeanette Winterson asks what we still stand to learn from the Suffragettes. Examining recent women's rights movements, the worlds of politics and technology, social media and changes in the law, Winterson celebrates how far we have come but demands that we do more. There is still so far to go, so much courage we still need to find. Witty and wise, incisive and inspiring, Courage Calls to Courage Everywhere is a powerful reminder of the need for true equality and an urgent call to arms.
A Sunday Telegraph Book of the Year `Winterson's winter tales unfailingly succeed in their endeavor to leave you aglow' - Guardian For years Jeanette Winterson has written a new story at Christmas time and here she brings together twelve of her brilliantly imaginative, funny, and bold tales, along with twelve delicious recipes for the twelve days of Christmas.
There are ghosts here and jovial spirits. Chances at love and tricks with time. There is frost and icicles, mistletoe and sledges. There's a cat and a dog and a solid silver frog. There's a Christmas cracker with a surprising gift inside. There's a haunted house and a SnowMama. There are Yuletides and holly wreaths. Three Kings. And a merry little Christmas time. And for the icing on the Christmas cake, there are twelve festive recipes from Yuletides past and present. Red cabbage, gravlax, turkey biryani, sherry trifle, Mrs Winterson's mince pies and more.
A baby girl is abandoned, banished from London to the storm-ravaged American city of New Bohemia. Her father has been driven mad by jealousy, her mother to exile by grief. Seventeen years later, Perdita doesn't know a lot about who she is or where she's come from - but she's about to find out.
'I saw the strangest sight tonight.' New Bohemia. America. A storm. A black man finds a white baby abandoned in the night. He gathers her up - light as a star - and decides to take her home. London. England. After the financial crash. Leo Kaiser knows how to make money but he doesn't know how to manage the jealousy he feels towards his best friend and his wife. Is the newborn baby even his? New Bohemia. 17 years later. A boy and a girl are falling in love but there's a lot they don't know about who they are and where they come from.
A 2013 World Book Night selection. Shortlisted for the Specsavers National Book Awards 'UK Author of the Year' 2012. A companion to Oranges are Not the Only Fruit, allowing Jeanette Winterson to explore the truth (often far worse than the fictionalised version) behind her autobiographical novel. The power of the written word to a child is brilliantly captured, books allowing the author to escape and find her own life – to write her own words.Like for Like ReadingBad Blood, Lorna SageOnce in a House on Fire, Andrea Ashworth
Featured on The Book Show on Sky Arts on 15 December 2011. Jeanette Winterson: In 1985 Jeanette Winterson's first novel, Oranges Are Not the Only Fruit , was published. It tells the story of a young girl adopted by Pentecostal parents. The girl is supposed to grow up and be a missionary. Instead she falls in love with a woman. Disaster. Written when Jeanette was only twenty-five, her novel went on to win the Whitbread First Novel award, become an international bestseller and inspire an award-winning BBC television adaptation. Oranges was semi-autobiographical. Mrs Winterson, a thwarted giantess, loomed over that novel and its author's life. When Jeanette finally left her home, at sixteen, because she was in love with a woman, Mrs Winterson asked her: why be happy when you could be normal? This book is the story of a life's work to find happiness. It is a book full of stories: about a girl locked out of her home, sitting on the doorstep all night; about a tyrant in place of a mother, who has two sets of false teeth and a revolver in the duster drawer, waiting for Armageddon; about growing up in an northern industrial town now changed beyond recognition, part of a community now vanished; and, about the Universe as a Cosmic Dustbin. It is the story of how the painful past Jeanette Winterson thought she had written over and repainted returned to haunt her later life, and sent her on a journey into madness and out again, in search of her real mother. It is also a book about other people's stories, showing how fiction and poetry can form a string of guiding lights, a life-raft which supports us when we are sinking.
'She is a master of her material, a writer in whom great talent abides' Vanity Fair
In this, her first collection of short stories, Winterson reveals all the facets of her extraordinary imagination. In prose that is full of imagery and word-play, she creates physical and psychological worlds that are at once familiar and yet shockingly strange.
These interlocking essays uncover art as an active force in the world - neither elitist or remote, present to those who want it, affecting even those who don't. Winterson's own passionate vision of art is presented here, provocatively and personally, in pieces on Modernism, autobiography, style, painting, the future of fiction, in two essays on Virginia Woolf, and more intimately in pieces where she describes her relationship to her work and the books that she loves.
'She is a master of her material, a writer in whom great talent abides' Vanity Fair


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Olathe & Southwest Joco
Olathe School District's 'ultimate show-and-tell'
The 'ultimate show-and-tell'
The Olathe School District calls it "the ultimate show-and-tell" for a fifth-grader as well as "a unique learning experience for all students" at Indian Creek Elementary School.
A fifth-grade girl's parents, aeronauts Jennifer and Maury Petrehn, brought one of their balloons to school this spring to demonstrate the science behind hot-air ballooning.
"During the week leading up to the hot-air balloon demonstration, our school focused their science lessons on standards such as states of matter (liquid, solid, gas), the Earth's spheres and weather patterns," said Principal Julie Sluyter. "Incorporating a hot-air balloon into those lessons provided a real-world experience for our students to see the science standards in action."
The special event began in the gym, where students learned about the history of hot-air ballooning. The flights date to the late 1700s when the Montgolfier brothers of France combined their interest in science with their jobs in the family paper business. One of their earliest experiments sent three small farm animals aloft in a basket suspended below a paper bag filled with hot air, the district said.
Today, hot-air balloons are made of coated nylon. Pilots initially fill them more than half full with cold air before turning on propane burners to heat the air. The hot air makes the balloon envelope rise, Jennifer Petrehn explained, and letting the air cool will cause the balloon to descend. The flight path is affected by the speed and direction of the wind because aeronauts can steer a balloon only by going up and down.
The students then went outside to see the inflation of a colorful balloon that held 56,000 cubic feet of air. That's smaller than the Petrehns' commercial balloon, which holds 105,000 cubic feet of air, the district said.
The students learned that hot-air ballooning is a nature-friendly form of travel that uses little fossil fuel.
"We combined our Earth Day activities that week and focused on how we can protect our environment," Sluyter said. "Hot-air ballooning is very protective of the environment and encourages outdoor activities."
Rabies clinic June 1 for Olathe residents
The Olathe Animal Shelter is holding a rabies vaccination clinic from 8 a.m. to noon June 1 at the Olathe Animal Shelter, 505 E. Sunvale Drive.
The clinic is open to Olathe residents only. The cost is $10 for a one-year vaccination and $30 for a vaccination that's effective for three years. Visitors can also take care of licensing their pets. Cash or check only.
For details on other services and prices, click on the calendar listing at olatheks.org.
Free summer lunches at Olathe Downtown Library
The Olathe Public Library is providing free meals on weekdays throughout the summer.
Through Aug. 14, kids can enjoy free lunches at the Olathe Downtown Library, 201 E. Park St., from 11 a.m. to 12:30 p.m. Mondays through Thursdays.
Meals are only for those ages 18 and under, but adults may accompany the children. Kindergartners and younger must be supervised. The meals, which must be eaten at the library, are sponsored by Harvesters Community Food Network, Friends of the Library and the Mayor's Christmas Tree Fund.
And on Fridays from June 7 through Aug. 9, free hot lunches will be offered for all ages from 11 a.m. to 1 p.m. at the downtown library. Menus will vary each week. The meals are presented in part by the Mayor's Christmas Tree Fund and Friends of the Library.
Registration is not required.
For a child's perspective on the past…
Every Wednesday and Thursday this summer, the Mahaffie Stagecoach Stop & Farm will offer "A Day Out with Frank and Ella," named for the two youngest Mahaffie children.
The historic site, at 1200 E. Kansas City Road, will explore a different theme each week related to growing up during the mid-1800s.
Weekday hours are 10 a.m. to 4 p.m. Admission is $5 per youth with one adult admitted free with each paid youth. A fee will be charged for additional adults.
Activities for June 5 and 6 focus on the pioneer trails and their history in this area. For the full schedule of themes, consult the calendar at mahaffie.org.
On Thursday nights in June and July — except for July 4 — the site will offer activities around the same themes during Family Fun Nights from 6 to 8 p.m. Admission to the night sessions are free, but there is a fee for stagecoach rides and some other activities.
Get rid of paint and chemicals
Residents from across Johnson County can dispose of old paint cans, gas and oil, chemicals, pesticides and other household hazardous waste at Olathe's drop-off location, which is open the second Saturday of each month.
The next free event will be from 8 a.m. to 2 p.m. June 8 at 1420 S. Robinson Drive. No appointment is necessary, but people should bring a photo ID and label items not in their original containers.
For a full list of acceptable items, go to olatheks.org.

subset: commoncrawl


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- This event has passed.
Monmouth Associates Luncheon
Thursday, February 21 @ 12:00 PM| $10
Event Navigation
At the next Monmouth Associates program on Feb. 21, educational studies professors Tammy La Prad, Michelle Holschuh Simmons and Craig Vivian will shed light on challenges faced by rural districts and also explain the College's new education initiatives.
The program will begin at noon in the Whiteman-McMillan Highlander Room of Stockdale Student Center.
Teaching at an institution located in a rural area, the College's educators are keenly aware of some rural school districts' struggle to recruit and retain teachers. In response to that need, the educational studies department has developed several initiatives, including its Rural Teacher Corps and TARTANS – Teachers Allied with Rural Towns and Neighborhood Schools.
The cost for a buffet lunch is $10 ($9 for Monmouth faculty and staff). Reservations can be made by calling 309-457-2231 by Feb. 19 or by email at email@example.com.
A free shuttle van transports passengers from two locations to every Associates luncheon. The van stops at the northeast quadrant of the Public Square at 11:45 a.m. and at the Faith United Presbyterian Church parking lot at around 11:50 a.m. It returns to both locations immediately following the program. Shuttle reservations can be made also by calling 309-457-2231.
Parking is available along North Ninth Street and in the parking lot near the Stockdale Center.


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Hotel Meurice
Property LocationWith a stay at Hotel Meurice in Calais (Calais City Center), you'll be steps from Musee des Beaux-Arts et de la Dentelle and 10 minutes by foot from Les Six Bourgeois. This hotel is 2.4 mi (3.9 km) from Port of Calais and 0.1 mi (0.2 km) from Eglise Notre Dame.RoomsStay in one of 39 guestrooms featuring flat-screen televisions. Your room comes with a pillowtop bed. Complimentary wireless Internet access is available to keep you connected. Conveniences include desks and coffee/tea makers, and you can also request cribs/infant beds (surcharge).AmenitiesTake in the views from a garden and make use of amenities such as complimentary wireless Internet access and gift shops/newsstands.DiningSatisfy your appetite for dinner at La Diligence, a restaurant which specializes in French cuisine, or stay in and take advantage of the room service (during limited hours). Quench your thirst with your favorite drink at the bar/lounge. Buffet breakfasts are available daily from 6:30 AM to 10:30 AM for a fee.Business, Other AmenitiesFeatured amenities include complimentary newspapers in the lobby, a 24-hour front desk, and luggage storage. Self parking (subject to charges) is available onsite.
Check in: 3:00 PM
Check Out: 11:30 AM
- 24-hour front desk
- Bar/lounge
- Breakfast available (surcharge)
- Elevator
- Free WiFi
- Free newspapers in lobby
- Free parking nearby
- Garden
- Gift shops or newsstand
- Laundry facilities
- Luggage storage
- Number of buildings/towers - 1
- Number of floors - 2
- One meeting room
- Parking (limited spaces)
- Safe-deposit box at front desk
- Self parking (surcharge)
- Total number of rooms - 39
- Year Built - 1771
Select room type
Hotel policy
Check-in Instructions
Government-issued photo identification and a credit card, debit card, or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
* All rates are per double room per night unless specified otherwise and exclude taxes and fees.
They are subject to availability. Some rates may require special identification. Extra people,
children, roll-away beds and incidentals may be subject to additional charges.
* Room pictures may differ from the actual room you receive.
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The flight of a military aircraft. View from the cockpit. - stock video
Your Easy-access (EZA) account allows those in your organisation to download content for the following uses:
- Tests
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It overrides the standard online composite licence for still images and video on the Getty Images website. The EZA account is not a licence. In order to finalise your project with the material you downloaded from your EZA account, you need to secure a licence. Without a licence, no further use can be made, such as:
- focus group presentations
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Because collections are continually updated, Getty Images cannot guarantee that any particular item will be available until time of licensing. Please carefully review any restrictions accompanying the Licensed Material on the Getty Images website and contact your Getty Images representative if you have a question about them. Your EZA account will remain in place for a year. Your Getty Images representative will discuss a renewal with you.
By clicking the Download button, you accept the responsibility for using unreleased content (including obtaining any clearances required for your use) and agree to abide by any restrictions.
- Advanced Tactical Fighter
- Aerial View
- Aeroplane
- Aeroplane Part
- Aerospace
- Aggression
- Air Force
- Air Raid
- Air Vehicle
- Aircraft Point of View
- Army
- Battle
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- Cockpit
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- Dashboard
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- Fighter
- Film
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- Military
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The tumor suppressor p53 is activated in response to many forms of cellular stress leading to cell cycle arrest, senescence or apoptosis. Appropriate sub-cellular localization is essential for modulating p53 function. We recently showed that p53 localizes to the nucleolus after proteasome inhibition with MG132 and this localization requires sequences within its carboxyl terminus. In the present study, we found that after treatment with MG132, p53 associates with a discrete sub-nucleolar component, the fibrillar center (FC), a region mainly enriched with RNA polymerase I. Moreover, we now demonstrate that this localization is an energy-dependent process as reduction of ATP levels prevents nucleolar localization. In addition, p53 sub-nucleolar accumulation is abolished when cells are subjected to various types of genotoxic stress. Furthermore, we show that monoubiquitination of p53, which causes it to localize to the cytoplasm and nucleoplasm, does not prevent the association of p53 with the nucleolus after MG132 treatment. Importantly, we demonstrate that p53 nucleolar association occurs in lung and bladder carcinomas.
The p53 tumor suppressor protein is a central target of inactivation in human cancer and a key regulator of genotoxic stress-induced growth arrest or apoptosis (Appella and Anderson, 2001; Giaccia and Kastan, 1998; Ko and Prives, 1996; Levine, 1997; Oren, 1994). Several reports have suggested a role for p53 in ribosome biogenesis (Budde and Grummt, 1999; Cairns and White, 1998; Chesnokov et al., 1996; Marechal et al., 1994; Sugimoto et al., 2003; Zhai and Comai, 2000). Mutations of the p53 core domain are one of the most frequent alterations detected in cancer cells (Levine, 1997). Interestingly, this region is needed to repress transcription by RNA polymerase I (Pol I) or RNA polymerase III (Pol III) (Stein et al., 2002a; Stein et al., 2002b; Zhai and Comai, 2000). Indeed, several point mutations in the core domain abrogate the capacity of p53 to repress rRNA synthesis (Budde and Grummt, 1999). Furthermore, in fibroblasts from p53 knock-out mice (p53–/–) the amount of pre-rRNA and tRNA is significantly higher than in wild-type animals (Budde and Grummt, 1999; Cairns and White, 1998). In accordance with these results p53-null epithelial cells have an elevated Pol I transcriptional activity compared to epithelial cells that express p53, indicating that repression of Pol I transcription by p53 occurs by an active repression mechanism (Zhai and Comai, 2000).
Multiple studies have demonstrated that under certain conditions p53 can be found in the nucleolus in several cell lines. The first study documenting p53 in the nucleolus revealed the presence of two different subsets of p53 protein in prostate and bladder carcinoma cells where the wild-type protein accumulates in the nucleolus whereas the mutant subgroup localizes to the nucleoplasm and is excluded from the nucleolus within the same cell (Benninghoff et al., 1999). p53 associates with nucleoli in detergent-permeabilized cells (Rubbi and Milner, 2000) and the proteasome inhibitor MG132 causes p53 to localize in the nucleolus and nucleoplasm (Karni-Schmidt et al., 2007; Klibanov et al., 2001; Latonen et al., 2003; Pokrovskaja et al., 2001). Nucleolar accumulation of p53 was also shown to be important for the reactivation of p53 in HeLa cells after cisplatin treatment (Horky et al., 2001; Wesierska-Gadek et al., 2002). Furthermore, in fibroblasts derived from spinal muscular atrophy (SMA) patients [caused by mutation in the survival motor neuron gene (SMN1)] with markedly reduced levels of SMN protein, p53 accumulated exclusively in the nucleolus (Young et al., 2002). Also, it has been reported that PRIMA-1, a compound that restores DNA binding properties and conformation of mutant p53, induces nucleolar translocation of mutant p53 (Rokaeus et al., 2006).
There are multiple potential mechanisms that might facilitate nucleolar localization of p53. p53 can bind sequences within the ribosomal gene cluster (Kern et al., 1991) and can associate with various nucleolar components such as L5 ribosomal protein (Marechal et al., 1994), topoisomerase I (Gobert et al., 1996), nucleolin (Daniely et al., 2002), nucleostemin (Tsai and McKay, 2002) and nucleophosmin (Colombo et al., 2002). Mdm2, the central negative regulator of p53 and an E3 ubiquitin ligase which ubiquitinates p53 and targets p53 for proteasome degradation, is not only able to form a complex with ribosomal proteins L5, L11 and L23 but itself is capable of nucleolar localization (Dai and Lu, 2004; Dai et al., 2004; Lohrum et al., 2003; Zhang et al., 2003). Nucleolar localization of Mdm2 and its association with the ribosomal proteins inhibit Mdm2 functions and help prevent p53 proteasomal degradation (Dai et al., 2004).
The nucleolus is the site of rRNA transcription, processing and ribosome assembly (Andersen et al., 2005; Carmo-Fonseca et al., 2000; Lam et al., 2005; Lamond and Sleeman, 2003; Leung et al., 2003; Olson and Dundr, 2005). As diagrammed in Fig. 1A the nucleolus consists of at least three morphologically distinct regions: the fibrillar center (FC), a dense fibrillar center (DFC) and a granular component (GC) (Scheer and Hock, 1999). Although, still not fully described, several studies indicate that the FC or the FC-DFC boundaries are the sites of pre-rRNA transcription (reviewed by Boisvert et al., 2007; Lamond and Sleeman, 2003). Ribosomal proteins and assembly factors are found in the GCs (Huang, 2002; Koberna et al., 2002; Lamond and Sleeman, 2003). FCs are particularly enriched in Pol I, topoisomerase I and the upstream binding factor (UBF, an activator of Pol I), whereas most processing molecules localize to the DFC and/or GC including nucleolin, nucleophosmin and fibrillarin [Casafont et al. (Casafont et al., 2007) and references therein].
In a previous study we showed that p53 accumulation in the nucleolus, after proteasome inhibition, requires two regions within the carboxyl terminus of p53 (Karni-Schmidt et al., 2007). We show now that after proteasome inhibition, p53 can associate with a discrete sub-nucleolar component, the FC. We demonstrate that p53 association with the FC is ATP dependent and it is abrogated by genotoxic stress. Our results also suggest that monoubiquitination of p53 does not preclude its association with nucleoli. Perhaps most relevantly we have found that p53 associates with nucleoli in human bladder and lung carcinomas.
p53 associates with a discrete sub-nucleolar compartment after MG132 treatment
To extend our observations and those of others that the treatment of cells with a proteasome inhibitor causes nucleolar localization of p53 (Karni-Schmidt et al., 2007; Klibanov et al., 2001; Latonen et al., 2003; Pokrovskaja et al., 2001) we examined the precise location of ectopic p53 within the nucleolus. H1299 cells were transfected with a construct expressing wild-type p53 and after 24 hours, cells were treated with different concentrations of MG132 for various times or left untreated. The levels of transfected p53 either did not change or were modestly reduced by treatment with MG132 in our system (Fig. 1B). Nucleolar localization of p53 could be detected by 8 hours after MG132 treatment (Fig. 1C). Surprisingly, when we compared the p53 staining pattern to that of nucleolin, a major nucleolar phosphoprotein that localizes to the DFC and GC (Bugler et al., 1982; Shaw and Jordan, 1995), nucleolin formed a visible ring around p53, indicating that in addition to weak or strong nucleoplasmic staining, p53 is localized in the FC following MG132 treatment (Fig. 1C). In cases where nucleoli contained more than one FC, p53 was detected in several FCs within the same nucleolus. DIC imaging confirmed our results, clearly showing p53 localization at the FC (Fig. 1C bottom row, E bottom row). In our experimental setting, the optimal concentration of MG132 was 10 μM, leading to approximately 10-15% of cells displaying nucleolar p53. However, in all cells with nucleolar p53, we found that p53 localized to the FCs (Fig. 1D). Using UBF as a well validated FC marker (Fig. 1E) (Casafont et al., 2007) we confirmed that the staining pattern with the UBF antibody is virtually identical to that observed with the p53 antibody after treatment with MG132. Furthermore, p53 localization to the FC following MG132 treatment was assessed in several cell lines expressing both wild-type and mutant forms of p53 (Table 1). Our results show that a subset of cell lines have detectable nucleolar p53.
Addition of the NoLS from the HIV-1 Rev protein to the p53 C terminus redirects p53 to the nucleolar DFC
We previously found that there are two discrete regions within the p53 C terminus that contribute to its ability to associate with nucleoli (Karni-Schmidt et al., 2007) although neither is homologous to the well characterized HIV-1 Rev NoLS (Hope, 1999; Meyer and Malim, 1994). HIV-1 Rev protein was shown to localize efficiently to nucleoli and to associate specifically with the nucleolar DFC and GC regions (Dundr et al., 1995). It was of interest to determine whether p53 nucleolar localization could be made more efficient by attaching it to the HIV-1 Rev NoLS. Moreover, we wanted to ascertain whether addition of a different well-defined NoLS to the p53 protein would have an impact on its association with the FCs portion of the nucleolus. To this end we compared nucleolar localization of p53 to that of p53 with the HIV-1 Rev NoLS attached to its C terminus (Rev NoLSC'p53; Fig. 2A). Constructs expressing either wild-type p53 or Rev NoLSC'p53 were introduced into H1299 cells with and without MG132 treatment. Levels of p53 protein were assessed by western blotting analysis (Fig. 2B). MG132 treatment again caused either no significant change or a modest reduction in ectopic p53 protein levels both with and without the Rev NoLS. By contrast, treatment with this inhibitor led to a marked increase in the levels of endogenous HSP70, in agreement with previous results showing induction of heat-shock response by proteasome inhibitors (Lee and Goldberg, 1998). At present we do not know the reason for the difference found in p53 and HSP70 levels in response to MG132, although we can conclude that the localization of p53 to the nucleolus after proteasome inhibition is not due to increased levels of p53. Strikingly, in contrast to the FC nucleolar localization of untagged wild-type p53 after MG132 treatment, the Rev NoLSC'p53 protein associated with the DFC-GC within the nucleolus. The expression levels of p53 and of Rev NoLSC'p53 are similar, hence, the difference in the sub-nucleolar localization is due to the differences in the NoLSs directing those proteins to the nucleolus. Furthermore, proteasome inhibitor treatment had no effect on the efficiency of localization of Rev NoLSC'p53, which was somewhat more than that of wild-type p53 (20–25%). Thus our results indicate that the HIV-1 Rev NoLS has a dominant effect over the regions in p53 required for p53 FCs localization. It is also of interest that whereas only 20–25% of the cells expressing p53 Rev NoLSC'p53 showed p53 accumulation in the nucleolus, full-length HIV-1 Rev protein could be detected within the nucleolus in virtually every cell (Fig. 3B and data not shown). Our data thus indicate that p53 possesses both determinants for nucleolar localization that differ from those of the HIV-1 Rev protein and regions that negatively regulate such association. Thus, we conclude that distinct partners or unique pathways are involved in MG132-induced p53 nucleolar localization.
p53 localization to the nucleolus occurs in an ATP-dependent manner and is abrogated by genotoxic stress
To determine whether genotoxic stress can impact sub-nucleolar accumulation of p53 and Rev NoLSC'p53, cells were treated with the topoisomerase II inhibitor and DNA intercalating agent, daunorubicin (D) and/or with MG132 (MD or M, respectively). Cells were either untreated (Fig. 3B panel 1) or treated with MG132 (Fig. 3B panel 2) or were treated with daunorubicin for 16 hours and then MG132 was added 8 hours before fixing the cells (Fig. 3B panel 3). Daunorubicin treatment diminished sub-nucleolar localization of untagged wild-type p53 and Rev NoLSC'p53 (Fig. 3B last panels, C). After daunorubicin treatment, nucleolin and Rev NoLSC'p53 localized primarily to the nucleoplasm or cytoplasm, respectively, whereas HIV-1 Rev protein localized to both the nucleolus and to the cytoplasm. Our results are consistent with previously described localization of HIV-1 Rev following treatment with α-amanitin, a Pol II and Pol III inhibitor or DRB, a Pol II inhibitor (Dundr et al., 1995). Furthermore, no p53 nucleolar localization was detected after MG132 treatment in cells that were pre-treated with either NCS (500 ng/ml) or UV [(25 J/m2), data not shown].
We previously discovered that ATP hydrolysis is required for p53 accumulation in the nucleolus in detergent-permeabilized cells (Karni-Schmidt et al., 2007). To assess the possible effect of reduction in ATP levels on p53 localization to the FCs after MG132 treatment, wild-type p53, HIV-1 Rev NoLSC'p53 and HIV-1 or Rev-GFP protein, as a positive control were transfected into H1299 cells, which were then treated with a combination of sodium azide (to inhibit cytochrome oxidase) and 2-deoxyglucose (to inhibit glycolysis) in order to reduce ATP levels in cells (Fig. 3A,B). Treatment with azide and 2-deoxyglucose (Az/DOG) had no effect on the overall levels of p53 and nucleolin (Fig. 3A, lanes a and Ma for cells treated with Az/DOG or MG132 and Az/DOG, respectively). Interestingly, treatment with Az/DOG eliminated MG132-induced p53 nucleolar association (Fig. 3B panel 5, C) whereas Rev NoLS-C'p53 still accumulated in the DFC-GC of cells treated with Az/DOG (Fig. 3B middle panel 5, C). HIV-1 Rev-GFP protein localization was also not affected by Az/DOG treatment (Fig. 3B panel 5). Thus the p53 NoLSs but not the Rev NoLS require ATP for nucleolar translocation. This result extends our findings that the Rev NoLS signal differs from that of p53 and confirms that the Rev NoLS has a dominant effect on the regions in p53 required for its nucleolar localization.
In summary, genotoxic stress had a significant inhibitory effect on the ability of wild-type p53 and Rev NoLSC'p53 but not intact HIV-1 Rev to localize to the nucleolus. p53 and Rev NoLS-C'p53 were both found to associate with the nucleoplasm whereas Rev protein associated both with nucleoli and the cytoplasm, and was excluded from the nucleoplasm. We cannot absolutely rule out that because of features present in HIV-1, the extensive nucleolar localization of Rev allows some of it to remain in the nucleolus. It is interesting that nucleolin, a well described nucleolar protein, behaves more like p53 in daunorubicin-treated cells.
Monoubiquitination of p53 does not preclude ectopic p53 from localizing to the nucleolus after MG132 treatment
The proteasome inhibitor MG132 can increase the steady state levels of ubiquitinated p53 in cells (Klibanov et al., 2001; Latonen et al., 2003; Maki et al., 1996). The primary E3 ligase for p53 is Mdm2 and monoubiquitination of p53 by Mdm2 has been reported to have profound consequences on p53 sub-cellular localization (Brooks and Gu, 2006; Li et al., 2003). To investigate a potential role of monoubiqitination on p53 nucleolar localization after treatment with proteasome inhibitor, we ectopically introduced p53 and p53 fused at its C terminus to a single ubiquitin molecule (p53-ubiqAA). We examined the localization of p53-ubiqAA with and without treatment with the proteasome inhibitor MG132. We found that in the absence of MG132 treatment p53-ubiAA was localized to the cytoplasm as well as the nucleoplasm as has been shown before (Li et al., 2003). Interestingly, however, after MG132 treatment p53-ubiqAA localized to the nucleolus and was associated with the FCs to the same extent as wild-type p53 (∼15%; Fig. 4). Thus, although we cannot conclude that monoubiquitination of p53 is required for its nucleolar localization, our data clearly show that monoubiquitinated p53 is not precluded from associating with nucleoli.
p53 is found associated with nucleoli in human tumor tissues
Previously, Benninghoff et al. reported that p53 can be detected in nucleoli of bladder and prostate cell lines (Benninghoff et al., 1999). As described above, p53 can be detected in the H1299 lung epithelial cancer cell line. It was therefore of interest to determine whether p53 can be detected in nucleoli from human bladder or lung carcinoma tissue sections. Indeed, in a subset of these tissues (five out of 48 bladder, five out of five lung), we detected p53 association with ∼10% of nucleoli in invasive areas, using immunohistochemistry (Fig. 5A). When we co-stained the tissues with anti-p53 and anti-UBF antibodies (as described in Materials and Methods) we detected co-localization of p53 and UBF by immunofluorescence (Fig. 5B). These experiments demonstrate that p53 can associate with nucleoli in some human invasive cancer subtypes.
In this study, we have shown that p53 localizes to a specific sub-nucleolar compartment (the FC) after proteasome inhibition. This localization is an active process that occurs in an ATP-dependent manner. Furthermore, we observed that treatment of cells with daunorubicin eliminated the association of p53 with the FCs of the nucleolus. Our results also show that the Rev NoLS differs from the nucleolar determinants on p53 in that the function of the Rev NoLS is unaffected by the proteasome inhibitor MG132 and it localizes within the GC and DFC of nucleoli. Furthermore, the Rev NoLS exerts a dominant effect when fused to p53. In addition, our results suggest that p53 monoubiquitination does not restrict p53 from associating with the nucleolus. Finally, we show that p53 localizes to the nucleolus in some invasive human lung and bladder carcinomas.
Multiple groups have reported that Mdm2-E3 ligase activity is crucial for both nuclear export and degradation of p53 (Boyd et al., 2000; Geyer et al., 2000; Haupt et al., 1997; Honda et al., 1997; Lohrum et al., 2001). Interestingly, Mdm2 primarily catalyzes the attachment of multiple single ubiquitin molecules to p53. Yet, the role of monoubiquitination on p53 is still a subject of investigation. At present, no transcriptional function for the monoubiquitination of p53 has been reported. Furthermore, monoubiquitinated p53 is not a substrate for the proteasome. This raises the question of whether this modification is necessary for p53 cytoplasmic accumulation. A study by the group of W. Gu showed that monoubiquitination of p53 by low levels of Mdm2 might be associated with p53 nuclear export, and high levels of Mdm2 induce polyubiquitination required for p53 degradation (Li et al., 2003). Additionally, these authors showed that a p53-ubiquitin fusion protein similar to the one used in our experiments that mimics monoubiquitinated p53 was found to be localized in the cytoplasm in an Mdm2-independent manner (Li et al., 2003). Other reports confirm that attachment of p53 to ubiquitin causes relocalization of p53 to the cytoplasm in both human and mouse cells (Joseph and Moll, 2003; Joseph et al., 2003). In our assay in which MG132 redirects p53 to nucleoli in a subset of cells, ectopic expression of p53 attached to ubiquitin did not prevent nucleolar localization of p53 after MG132 treatment. Since we could not detect significant amounts of a slower migrating form of p53 after MG132 treatment, this suggest that only a minor amount of the steady state level of p53 in our experiments is monoubiquitinated after this treatment. If that is the case then we suggest that ubiquitin is not required for p53 nucleolar localization, nor does it prevent it. We cannot rule out, however, that ectopic expression of wild-type p53 in H1299 cells results in monoubiquitination of p53 as the levels of endogenous Mdm2 induced by p53 are insufficient for p53 degradation and it is the monoubiquitinated form of p53 that is found associated with nucleoli. Further experiments will be required in order to clarify the role of ubiquitin, if any, in the association of p53 with nucleoli.
Under stress conditions such as proteasomal inhibition where multiple proteins cannot be degraded, limiting ribosomal biosynthesis might be beneficial to the cell and important for its survival. Regulating rRNA biogenesis could be an important step in regulation of ribogenesis. We observed p53 to be associated with the FCs of the nucleolus, a sub-nucleolar domain enriched in Pol I, the polymerase responsible for rRNA synthesis. Indeed, p53 has been shown to repress Pol I transcription in vitro (Budde and Grummt, 1999; Zhai and Comai, 2000). Interestingly, transcription experiments with p53 deletion mutants have indicated that the p53 carboxyl terminus is essential, but not sufficient, to repress transcription of Pol I (Zhai and Comai, 2000). This is in agreement with our previous study showing that the p53 carboxyl terminus is crucial for p53 nucleolar localization (Karni-Schmidt et al., 2007). Our results here suggest another link between p53, Pol I and ribosomal biosynthesis. It is tempting to speculate that p53 localizes to the FCs of the nucleolus in order to repress Pol I. Further investigation of the direct effect of p53 on Pol I followed by MG132 treatment may provide a greater insight into p53-mediated regulation of Pol I. Perhaps regulation of ribosomal biogenesis could be explored as yet another target for therapeutic intervention in cells with deregulated cell proliferation.
It is interesting that in both cell lines and tissues we detected approximately only 10% of cells with nucleolar p53. Although we do not, as yet, have an explanation for this distribution, we can offer some conjectures. First, p53 nucleolar localization may be cell cycle stage dependent. Second, the conformation of p53 protein may play a role as previously shown by others (Benninghoff et al., 1999; Rokaeus et al., 2006). Third, p53 modifications may regulate its association with nucleoli as we previously showed (Karni-Schmidt et al., 2007). These and other possible scenarios may also explain why no nucleolar p53 is detected in some cell lines, as listed in Table 1. It is noteworthy that whereas HIV-1 Rev protein displays highly efficient nucleolar association, p53 fused to HIV-1 Rev NoLS nucleolar localization is a much less frequent event (Fig. 2D, Fig. 3C). Elucidating the mode(s) of p53 nucleolar localization will be a direction for future studies.
Materials and Methods
Cell culture and treatments
IMR90, LNCAP, WI38, U2OS and HCT-116 cells were grown and maintained in DME medium supplemented with 10% fetal bovine serum. H1299 cells (p53-null) expressing tetracycline-regulated wild-type p53, or several tumor-derived mutant forms of p53, including p53H175, p53S249, p53A143 and p53S248 were generated as previously described (Chen et al., 1996; Resnitzky et al., 1994). Cells were grown and maintained in DME medium supplemented with 10% fetal bovine serum, puromycin (2 μg/ml; Sigma), G418 (300 μg/ml; Invitrogen), and tetracycline (4.5 μg/ml; Sigma). To induce p53, cultures were washed twice with tetracycline-free medium following plating in the above medium lacking tetracycline for 24 hours. HT-29 cells (p53H273) were grown in McCoy medium supplemented with 10% fetal bovine serum, and T98G (p53I237) cells were grown in MEM with 2 mM L-glutamine supplemented with 10% fetal bovine serum. To inhibit proteasome function, cells were incubated with MG132 (10 μM) for 8 hours or as described in the text. For ATP depletion experiments, cells were washed twice with PBS and were then incubated for 45 minutes at room temperature with 5 mM sodium azide (NaN3) and 1 mM 2-deoxyglucose. To induce cellular stress, cells were treated with 0.22 μM daunorubicin (Sigma) for 16 hours.
Western blotting
Cells were washed twice with PBS and then collected by scraping into cold extraction buffer [10 mM Tris (pH 7.5), 1 mM EDTA, 400 mM NaCl, 10% glycerol, 0.5% NP40, 5 mM NaF, 1 mM DTT and 0.1 mM PMSF] followed by centrifugation at 13,000 rpm for 10 minutes at 4°C in a tabletop centrifuge. Supernatants were resuspened in 3× protein sample buffer [30% glycerol, 2 M β-mercaptoethanol, 12% SDS, 500 mM Tris (pH 6.8) and 0.5 mg/ml Bromophenol Blue] and boiled for 10 minutes at 95°C. The samples were loaded onto SDS-polyacrylamide gels and transferred to nitrocellulose for 3 hours. SDS-PAGE and immunoblotting procedures were performed under standard conditions. Monoclonal anti-p53 antibodies PAb 1801 and PAb DO-1 were used to detect p53. The anti-nucleolin antibody was used as described previously (Ghisolfi-Nieto et al., 1996). An anti-actin antibody (Sigma) was used as a loading control and anti-HSP70 (Santa Cruz Biotechnology) was used as a control to proteasome inhibition treatment.
Immunofluorescence in cell lines
Fourty-eight hours after plating in 60 mm culture dishes, cells were washed twice with PBS followed by incubation with 4% paraformaldehyde (Sigma) for 15 minutes. Cells were then washed three times with PBS, treated with PBS plus 0.5% Triton X-100 for 1.5 minutes and blocked with 0.5% BSA (Sigma) in PBS for 30 minutes at room temperature prior to incubation with 50 μl of the diluted primary antibodies, PAb DO-1, PAb 1801 and anti-nucleolin or anti-UBF (Santa Cruz) as indicated for 1 hour at room temperature. The coverslips were then washed three times with PBS and double staining was performed by incubating the coverslips for 1 hour with 50 μl of diluted (1:100) secondary Cy5 goat anti-mouse IgG antibody (Jackson Immunoresearch) and Alexa Fluor® 488 goat anti-rabbit (Molecular Probes). Coverslips were washed three times with PBS and mounted with 10 μl cold 50% glycerol. The images were collected and analyzed using confocal laser scanning microscopy (Olympus, model 1X70) and Fluoview software. In order to directly visualize nuclei and nucleoli, differential interference contrast (DIC) images were taken in parallel. To quantify the data, p53 proteins were analyzed by visually screening fields of 200 cells or more and choosing representative images. Cells were scored as positive for p53 nucleolar localization if p53 was detected in one or more nucleoli.
Immunohistochemistry and immunofluorescence in human tissues
Immunohistochemical and immunofluorescence analyses were conducted on formalin-fixed and paraffin-embedded tissue sections from human lung and bladder carcinomas. Sections (5 μm) were de-paraffinized and submitted to antigen retrieval by steam treatment for 15 minutes in 10 mM citrate buffer at pH 6.0. Subsequently, slides were incubated in 10% normal horse serum (for immunohistochemistry) or normal donkey serum (for immunofluorescence) for 30 minutes, followed by incubation with primary antibody overnight at 4°C. The primary antibodies used were mouse monoclonal anti-p53 (PAb1801; Calciochem) for immunohistochemistry, and rabbit polyclonal anti-p53 (Santa Cruz) and mouse monoclonal anti-UBF (Santa Cruz) for immunofluorescence. Slides were then incubated with Alexa Fluor® 488 anti-rabbit and Alexa Fluor® 594 anti-mouse for 30 minutes (Invitrogen) and then mounted with Mounting Medium for Fluorescence (Vector Laboratories, Inc.). For immunohistochemistry, slides were incubated with biotinylated anti-mouse immunoglobulins for 30 minutes (Vector Laboratories, Inc.) followed by avidin-biotin peroxidase complexes (Vector Laboratories, Inc.) for 30 minutes. Diaminobenzidine was used as chromogen and Hematoxylin as the nuclear counter-stain. The immunohistochemistry images were collected using a standard light microscope and the immunofluorescence images were collected and analyzed as stated above using confocal laser scanning microscopy (Olympus model 1X70) and Fluoview software.
Transfections and plasmids
Transfections were performed using Lipofectamine 2000 reagent (Invitrogen) in accordance with the manufacturer's protocols. Expression vectors derived from pcDNA3 were pc53-expressing full-length p53 cDNA and Rev NoLS-C'p53, expressing full-length p53 with the nucleolar localization signal (NoLS) of HIV-1 Rev protein attached to its carboxyl terminus. The Rev NoLS-C'p53 construct was made using standard cloning procedures and was confirmed by sequencing. Rev-GFP was a generous gift from Dirk Daelemans (Rega Institute for Medical Research, Belgium) (Stauber et al., 1998).
A construct expressing p53 fused to ubiquitin at the p53 carboxyl terminus (p53-ubiAA) was generously provided by Wei Gu (Columbia University, New York, NY) and made as described by Li et al. except that the last two amino acids of ubiquitin were mutated to alanine (Li et al., 2003). We are grateful to all the members of the Prives lab and particularly to Ella Freulich, Masha Poyurovsky, Nicole Baptiste, Lynn Biderman, Yingchun Li, Yan Zhu and Igor Matushansky for technical assistance, valuable discussions and great suggestions. This work was supported by grants CA58316, CA87497 and CA009503 from the NIH.
- Accepted August 17, 2008.
- © The Company of Biologists Limited 2008


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After two successive months of weather-imposed cancellations, there will be a Poetry Hoot on Wednesday, April 4.
The Hoot is held at Cafe Espresso in the 800 Plaza on Islington Street in Portsmouth, beginning usually a little before 7 p.m.
The featured poets on April 4th will be Lee Sharkey and Adam Scheffler
Sharkey�s poetry collections are "Walking Backwards" (Tupelo Press, 2016), "Calendars of Fire" (Tupelo, 2013), "A Darker, Sweeter String" (Off the Grid, 2008), and eight other full-length poetry books and chapbooks. Her work has appeared in Crazyhorse, FIELD, Kenyon Review, Massachusetts Review, Nimrod, Pleiades, Seattle Review, and other journals.
Her recognitions include the Ballymaloe International Poetry Prize, the Abraham Sutzkever Centennial Translation Prize, the Maine Arts Commission�s Fellowship in Literary Arts, the Maine Writers and Publishers Alliance�s Distinguished Achievement Award, and the RHINO Editor�s Prize. She leads a creative writing workshop for adults recovering from mental illness and serves as Senior Editor of the Beloit Poetry Journal.
Scheffler received his MFA in poetry from the Iowa Writers� Workshop, and his PhD in English from Harvard. His first book of poems � "A Dog�s Life" � was selected by Denise Duhamel as the winner of the Jacar Press Poetry Book Contest. His poems have appeared in The American Poetry Review, The Antioch Review, Rattle, North American Review, Verse Daily, and many other venues. He has been nominated three times for the Pushcart Prize and is the winner of River Styx�s 2014 International Poetry Contest.
Doors open about 6 p.m. at Cafe Espresso, 738 Islington St. (in the 800 Islington Plaza), Portsmouth. An �open mic� immediately follows the featured poets so bring your poem and be ready to read. Please limit your poem to 1 page or approx. 2 minutes. Please: no announcements from the podium. For more information, visit www.pplp.org.


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We consider the Hassan-Rosen bimetric field equations in vacuum when the two metrics share a single common null direction in a spherically symmetric configuration. By solving these equations, we obtain a class of exact solutions of the generalized Vaidya type parametrized by an arbitrary function. Besides not being asymptotically flat, the found solutions are nonstationary admitting only three global spacelike Killing vector fields which are the generators of spatial rotations. Hence, these are spherically symmetric bimetric vacuum solutions with the minimal number of isometries. The absence of staticity formally disproves an analogue statement to Birkhoff's theorem in the ghost-free bimetric theory which would state that a spherically symmetric solution is necessarily static in empty space.
The ghost-free bimetric theory is an extension to general relativity where two metric tensors are used instead of one. A priori, the two metrics may not have compatible notions of space and time, which makes the formulation of the initial-value problem problematic. Moreover, the metrics are coupled through a specific ghost-free interaction term that is nonunique and possibly nonreal. We prove that the reality of the bimetric potential leads to a classification of the allowed configurations of the two metrics in terms of the intersections of their null cones. Then, the equations of motion and general covariance are enough to restrict down the allowed configurations to metrics that admit compatible notions of space and time, and furthermore, lead to a unique definition of the potential. This ensures that the ghost-free bimetric theory can be defined unambiguously. In addition, we apply the results on spherically symmetric spacetimes. First, we explore the behavior of the black hole solutions both at the common Killing horizon and at the large radii. The study leads to a new classification for black holes within the bidiagonal ansatz. Finally, we consider the bimetric field equations in vacuum when the two metrics share a single common null direction. We obtain a class of exact solutions of the generalized Vaidya type parametrized by an arbitrary function. The found solutions are nonstationary and thus nonstatic, which formally disproves an analogous statement to Birkhoff's theorem in the ghost-free bimetric theory.
The Hassan–Rosen bimetric theory is an extension of general relativity which considers the interaction between two metric fields defined on the same differentiable manifold. Self-accelerating cosmologies are exact solutions of this theory, and this makes it interesting to explore. We analyze the theory and see if it can provide other physically consistent solutions.
Despite the effort put in studying the theory in recent years, very few exact solutions are known in the literature, and the majority are equivalent to those of general relativity. A valuable approach to try to simplify the field equations and find exact solutions is to impose some spacetime symmetry on the system, e.g., spherical symmetry.
Our study is concerned with symmetries of spacetimes in Hassan–Rosen bimetric theory. Two metrics being present, we investigate what relations exist between their spacetime symmetries. We focus on the isometries of the metrics and clarify when they are the same.
We apply the results in exploring solutions in the Hassan–Rosen theory. We consider maximally symmetric solutions and black hole solutions, and find a previously unknown class of non-stationary spherically symmetric solutions. The existence of the class of non-stationary spherically symmetric solutions disproof a similar statement to Birkhoff's theorem in the Hassan–Rosen bimetric theory. The study of bidiagonal non-rotating black holes sharing the isometries focuses on their properties both at the shared Killing horizon and far from it.
The thesis is a review of these results, and the relevant papers accompany it.


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#132 - 315 Dickson Cres, Saskatoon
The "Woodbridge Condominiums" at The Willows. Care-free lifestyle with all the features you want!! Bright south facing exposure with an incredible panoramic view of the Willows golf course and clubhouse from the Kitchen, dining room and living room areas. Plenty of room in this 1,769 sq. ft. condo with 9' ceilings, hardwood flooring and in-floor heating. Two spacious bedrooms plus a den. Walk-in closet off Master Bedroom plus 4 piece ensuite with separate tub and shower; Storage and laundry room with washer and dryer; granite kitchen counter tops; garden door to deck with natural gas hookup for the barbecue. Quality custom window blinds included. Amenities room with visiting area, shuffle board and exercise room. One heated underground parking stall, one surface parking stall (Two underground parking stalls available if desired) plus large storage locker and carwash bay.Lots of visitor parking! Enjoy the convenience of the golf course, Stonebridge shopping facilities and quick access to downtown. You will be impressed!! SELLER WILL CONSIDER TRADE!........... PUBLIC Open House: 2:00pm - 4:00pm Saturday & Sunday, September 21st & 22nd, 2019.
Amenities Room, Elevator, Exercise Area
Air Conditioner (Central), Elevator, Intercom, Natural Gas Bbq Hookup, Wheelchair Accessible
Dryer, Dishwasher Built In, Fridge, Garburator, Microwave Hood Fan, Stove, Window Treatment, Washer
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The above information is from sources deemed reliable but should not be relied upon without independent verification. The information presented here is for general interest only, no guarantees apply.
MLS®, REALTOR®, and the associated logos are trademarks of The Canadian Real Estate Association.


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Story map: Seeking softer shores on Puget Sound
A 2019 story map produced by the University of Washington Puget Sound Institute in collaboration with the Washington Department of Fish and Wildlife shows how shoreline armoring can often be replaced by softer, shore-friendly features.
- Action Agenda
- Adaptive management
- Algae
- Amphibians
- Aquatic reserves
- Bald eagles
- Ballard Locks
- Biennial Science Work Plan
- Biofilm
- Birds
- Bivalves
- Circulation
- Climate change
- Contaminants of emerging concern
- Counties
- Disease
- Dungeness crabs
- Ecoregions
- Ecosystem-based management
- Ecosystem services
- Eelgrass
- Estuarine habitat
- Eyes Over Puget Sound
- Fishes
- Floodplains
- Food web
- Forage fish
- Freshwater habitat
- Geography
- Harbor porpoise
- Harbor seals
- Harmful algal blooms
- Healthy human population
- Herring
- History
- Human quality of life
- Hypoxia
- Implementation Strategies
- Invasive species
- Invertebrates
- Jellyfish
- Kelp
- Killer whales
- Mammals
- Marine birds
- Marine debris
- Marine habitat
- Marine Protected Areas
- Marine Waters Overview
- Modeling
- Monitoring
- National Estuary Program
- Nearshore habitat
- Noise
- Nutrient pollution
- Ocean acidification
- Oil spills
- Persistent contaminants
- Physical environment
- Plants
- Puget Sound boundaries
- Puget Sound Fact Book
- Puget Sound Pressures Assessment
- Puget Sound Update
- Reptiles
- Rockfish
- Salish Sea
- Salish Sea Currents magazine
- Salish Sea Ecosystem Conference
- Salmonids
- Sea-star wasting disease
- Sea level rise
- Sewage and fecal pollution
- Shellfish
- Shoreline armoring
- Social science
- Species and food webs
- Species of concern
- State of the Sound
- Stormwater
- Terrestrial habitat
- Tidal energy
- Toxic contaminants
- Traditional ecological knowledge
- Tribes
- Vital Signs
- Water quality
- Water quantity
- Water Resource Inventory Areas (WRIA)
- Watershed hydrologic unit codes (HUC)
- Watersheds
- Zooplankton
Connections between terrestrial, freshwater, and marine habitats
Freshwater habitat in the Puget Sound region consists of rivers, marshes, streams, lakes and ponds that do not have any saltwater input. Many species depend on these freshwater resources, including salmon, salamanders, frogs, and beavers.
Eyes Over Puget Sound: Surface Conditions Report - July 29, 2019
In July, the recent trends of warm, dry conditions lessened; however, river flows remain low. Extensive macroalgae drifted through South and Central Sound and washed up on beaches. Macroalgae growth is fueled by excessive nutrients and sunshine. When it washes onto the beach, it is called beach wrack, and it can be a health risk to beachgoers because of bacteria it can harbor. From our aerial photography, we saw that Southern Hood Canal looks tropical because of a bloom of coccolithophores coloring the water turquoise. Schools of fish congregate in South Sound and southern Hood Canal. Jellyfish are abundant in Quartermaster Harbor.
Ancient harvests: A history of Salish Sea herring
Scientists believe that herring have been a staple of Salish Sea food and culture since humans first arrived here at least 12,500 years ago. That importance has continued into modern times, even as herring numbers have declined in parts of the region.
Salish Sea Model looks at climate impacts on the nearshore
A 2019 paper in the Journal of Geophysical Research: Oceans outlines how the Salish Sea Model describes the impacts of climate change, sea level rise and nutrient loads on the region's nearshore environment.
Unsung seabirds could help track Puget Sound health
Pigeon guillemots have attracted relatively little scientific attention compared to other seabirds in Puget Sound. That may be because their population is generally stable, but a group of citizen scientists is helping to put guillemots on the conservation radar. They hope the birds can be used as an indicator of Puget Sound health.
Fine-scale variability in harbor seal foraging behavior
A 2014 paper in the journal PLoS ONE examines differences between foraging behavior of harbor seals based on haulout site locations, seasons, sexes and times of day. The authors hypothesize that these factors may help explain the variability in diet among harbor seals observed at different haul-out site groups in the Salish Sea.
Eyes Over Puget Sound: Surface Conditions Report - May/June 2019
Warm and dry conditions this spring are predicted to persist into summer, resulting in saltier and warmer than normal Puget Sound water conditions. Early upwelling and a premature melt of the snowpack means nutrient-rich ocean water likely already entered Puget Sound. This sets the stage for a lot of biological activity. From the air, it is obvious that the productive season is in full swing. We saw large algae blooms in Central Sound along with abundant Noctiluca. Huge numbers of anchovies were documented in Case Inlet and other finger inlets in South Sound, attracting hundreds of marine mammals.
Video series features science and adventure in the Salish Sea
A new video series follows local scientists into the water, capturing the adventure behind the research. "Salish Sea Wild" is entering its second season and we interviewed the series host and producers. Among our burning questions: What's it like to have a Steller sea lion chew on your head?
Predator–prey dynamics of bald eagles and glaucous‐winged gulls
An unintended consequence of the recovery of bald eagles (Haliaeetus leucocephalus) has been the impact on seabirds. The authors of a 2019 paper published in Ecology and Evolution suggest that the effects of bald eagle activity on a large glaucous-winged gull (Larus glaucescens) colony on Protection Island in the Strait of Juan de Fuca include the possibility of coexistence but also the possibility of gull colony extinction.
Science in the spotlight: Eelgrass recovery
The Washington Department of Natural Resources is studying new ways of increasing ecologically important eelgrass habitat in Puget Sound. It is part of the state's effort to boost eelgrass 20% Sound-wide by 2020. So far, recovery of the species has fallen short of that goal, but transplanting efforts are showing promise.
State aquatic reserves lean heavily on citizen scientists
Eight aquatic reserves in Puget Sound are being studied by volunteers working under the direction of state experts. Washington Department of Natural Resources manages the reserves with guidance from nearby communities.
Story map: European green crab in Puget Sound
A May 2019 story map shows how a broad collaboration of volunteers, agencies, and tribes is working together to keep invasive European green crabs at bay in Washington state. The story map was produced by the Encyclopedia of Puget Sound and the Puget Sound Institute in collaboration with the Washington Sea Grant Crab Team.
Washington state aquatic reserves
Eight aquatic reserves, managed by the Washington Department of Natural Resources, have been established to protect important ecosystems on state aquatic lands.
In most reserves, area residents work with state, local and tribal officials and nonprofit groups to develop and carry out management plans, including scientific research.
The aquatic reserves in the order they were established include:
- Maury Island 2004
- Cypress Island 2007
- Fidalgo Bay 2008
- Cherry Point 2010
- Protection Island 2010
- Smith and Minor Islands 2010
- Nisqually Reach 2011
- Lake Kapowsin 2016
Survey illustrates a lack of familiarity with the Salish Sea
Washington and British Columbia residents are largely unfamiliar with the Salish Sea. A recent study conducted by the SeaDoc Society and Oregon State University reveals a need to improve geographic literacy and familiarity with the Salish Sea among those communities who share and live alongside this integrated transboundary ecosystem. This summary was provided by two of the collaborators on the survey, David Trimbach of Oregon State University and Joe Gaydos, Science Director at the SeaDoc Society.
Ghost nets still fishing in the deep waters of Puget Sound
New technology is helping to remove deadly "ghost nets" that have been lost in the depths of Puget Sound. It is part of an effort that saves millions of animals every year, but managers say better reporting of these lost nets by fishermen is still needed.
The herring defenders
Each winter and spring, researchers survey the sometimes spectacular spawning events of Puget Sound's Pacific herring. They have found wide swings in the fish's population and an overall decline in herring numbers since the 1970s, but little is known about the cause or what this might mean for the health of the food web. We spent a day with a biologist spotting herring eggs and considering the future of one of our region's most ecologically and culturally important fish species.
Toxics in Fish Implementation Strategy
The Toxics in Fish Implementation Strategy is a recovery plan that will be used to guide funding and activities to reduce the impacts of toxics contaminants on marine fish and the humans that consume them. The plan is scheduled to be completed in 2019.
Eyes Over Puget Sound: Surface Conditions Report - March 26
Following a generally warmer and drier winter and then a cold spell in February, Puget Sound waters are cold for anchovies. The productive season is in full swing with algal blooms spotted in South Sound, Kitsap Peninsula, and Quartermaster Harbor. Jellyfish are abundant in some inlets, and Noctiluca stains the water orange in Hood Canal. We collect monthly data to keep you informed about the conditions around Puget Sound. Come into the lab and see how we assure the highest data quality from our field instruments!
Social networks a key to orca survival
Understanding the social networks and family bonds of Puget Sound's southern resident orcas may be critical to keeping the endangered whales from extinction. A healthy population is about more than numbers, scientists say. It's about connections.
2018 Salish Sea toxics monitoring synthesis: A selection of research
A 2019 report from the Puget Sound Ecosystem Monitoring Program presents an overview of selected recent monitoring and research activities focused on toxic contaminants in the Salish Sea.
Tracking the effectiveness of armor removal in Puget Sound
The removal of shoreline armoring has become a priority for the state's Puget Sound recovery efforts. The Puget Sound Ecosystem Monitoring Program (PSEMP) is providing details on the effectiveness of armor-removal projects at 49 study sites.
Risk of a major oil spill generates action in Olympia
Bills in the state legislature target oil spill threats to Puget Sound and its endangered killer whales.
Oil spill risks by the numbers
An EPA-funded study of oil spill risks in Puget Sound forms the basis of new legislation to regulate vessel traffic in the region. We break down some of the numbers from the study and look at where the risks may be greatest.
The survival of hatchery‐origin pinto abalone Haliotis kamtschatkana released into Washington waters
In Washington State, the pinto abalone (Haliotis kamtschatkana) has declined by 97 percent since 1992 and is unlikely to recover without intervention. A captive rearing and restocking pilot study shows promise for saving wild populations from local extinction.
Disease epidemic and a marine heat wave are associated with the continental-scale collapse of a pivotal predator (Pycnopodia helianthoides)
The sunflower sea star (Pycnopodia helianthoides) is highly susceptible to sea star wasting disease. The authors of a 2019 paper published in Science Advances document the rapid, widespread decline of sunflower stars and discuss the ecological implications of losing this important subtidal predator species.
The Orca Docs: Can medical interventions help?
This three-part series explores opportunities and challenges of using medical interventions to save Puget Sound's southern resident orcas from extinction. Part 1 looks at how scientists might treat endangered southern resident orcas that face starvation and risks of disease; Part 2 considers how veterinarians have intervened with other animals in the wild, and how this might apply to orcas in Puget Sound; and Part 3 explores a federally approved vaccination program designed to ward of a deadly virus among endangered Hawaiian monk seals.
Eyes Over Puget Sound: Surface Conditions Report - February 21, 2019
Although fall and winter were warm, February brought cold snowy weather and low river flows. Despite colder air temperatures, the productive season has already started in Hood Canal and Holmes Harbor. Puget Sound waters were warmer than expected through January, and the warmest waters were in Hood Canal, possibly creating a thermal refuge for cold-sensitive species such as anchovies. We saw lots of sea lions feasting on anchovies in Case Inlet, and we may have captured some herring spawning activity. Unusual for mid-winter, we saw jellyfish patches in Eld and Budd inlets. See the new publication about ocean acidification featuring twenty-five years of our marine monitoring data!
Return of a native: Olympia oysters are making a comeback
Puget Sound's only native oysters were nearly wiped out in the 19th century from overharvesting. Now a network of scientists and advocates is working to restore them to their historical and cultural prominence.
The whales in our waters: The economic contribution of whale watching in San Juan County
A 2019 report from the non-profit group Earth Economics look at revenues and other economic activity resulting from whale watching in San Juan County, Washington.
Vaccines now used to reduce the risk of extinction in Hawaiian monk seals
For critically endangered animal populations, experts worry that a highly infectious disease could be the final nail in the coffin, forcing the species into extinction. That's one reason why federal authorities approved the development and deployment of a new vaccine to ward off the deadly morbillivirus among Hawaiian monk seals. The vaccination program raises the possibility of using vaccines to prevent disease among Puget Sound's southern resident killer whales, but no specific steps have been taken so far.


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The Brave One (R1) in February
Warner Home Video have announced the Region 1 DVD release of The Brave One on 5th February 2008 priced at $28.98 SRP. Jodie Foster stars as Erica Bain, host of a New York radio show whose life is shattered by a brutal attack that leaves her fiancé (Naveen Andrews, Lost) dead, her own body shattered and her sense of safety permanently destroyed. Now, when Erica's fear of her beloved city becomes too much to bear, she arms herself...
Directed by Neil Jordan, this vigilante thriller also stars Terence Howard.
Available in separate Widescreen and Full Screen editions, features include:
- English DD5.1 Surround
- English, French and Spanish subtitles
- Featurette I Walk The City (22 mins)
- Additional Scenes (7 mins)


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Tech and the City: MOU with UN Habitat Aims To Improve Urban Planning in South Asia
New York, USA – A Memorandum of Understanding (MoU) has been reached between the UN Office of Communication and Information Technologies (OICT) and UN-Habitat that provides a framework of cooperation regarding the development of emerging technology tools and digital platforms for urban design and planning. Read More.
The creation of new technologies for urban planning and municipal governance is a reality which improves the provision of services to citizens. ICT and UN-Habitat are pleased to join efforts with the goal of fostering inclusive, safe, resilient and sustainable cities, as enshrined in Agenda 2030 for Sustainable Development.
The MoU identifies four principle areas of collaboration: 1) Facilitate the development and implementation of emerging technology tools and digital technologies to assist in building capacity for safe and resilient cities; 2) Jointly monitor the development of emerging technology tools and digital technologies related to sustainable urbanization; 3) Collaborate and participate in events of mutual interest; 4) Strengthen collaborations through joint public/private partnerships at the regional and country level with a focus on Afghanistan, India, Bangladesh, and Sri Lanka.
UN Habitat is the focal point for sustainable urbanization in the UN system. With more than 40 years of experience worldwide, this programme works in over 90 member states to create a better urban future for all as more than half of the world population is already living in cities. Its core mission is to promote the development of socially and environmentally sustainable human settlements and strive for adequate shelter with better living standards for all.
The MoU with Habitat is the latest MoU signed between OICT and UN entities. Last month an MOU was reached with UNICRI. Together with its emerging network of UNTIL Labs – which operate in four countries – OICT is emerging as a go-to source for UN bodies and member states seeking technology solutions.


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If you have purchased an iPhone from a friend or second hand, you could be experiencing problems with your iCloud.
iCloud is one of many ways to sync, store, and update data between Apple devices. You are allowed 5 GB of free iCloud space but it can fill quite fast when adding photos and documents. iCloud is very useful when you want to free up space on your handset. You can decide what data is backed up and what is not. For example, you can sync photos, and documents and apps, but you don't have to if you do not wish to. It might be wise to back your data up to your computer as well.
If you have received a phone with iCloud lock, this means that the previous owner is still connected to that handset through iCloud and before you can use it, iCloud needs to be unlocked. iCloud is similar to an external hard drive, but instead of a physical item holding your data, everything is accounted for online.
Whenever you purchase a new iPhone from either your network or from the Apple Store, your phone is in its most secure state. After setting up your phone, including your iCloud with your Apple ID and password, you have secured your phone to you and only you as the user. This is called an activation lock. It makes it harder for anyone else to use your phone if it's ever lost or stolen.
Therefore, when you purchase a phone from someone else, the phone should be reset back to its factory settings in order to unlock iCloud for the next user. But if this is not the case when you purchased your second hand phone, we can help you with that.
Official iPhone unlock offers the service of unlocking your iCloud if it is locked from the previous owner and still has information data stored on the phone. Unfortunately, you will not be able to use your phone if the iCloud lock is activated.
You can easily unlock your iPhone or other device such as an iPad with us by simply entering your IMEI or serial number. This will reset the device's iCloud and you will then be free to enter your own Apple ID and password.


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Breaking news from around the world
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Is there any planning for the deprecated features that were announced prior to version 9.x and when they will definitely stop working?
For example Dialogs, some of the client APIs, or in general:social.technet.microsoft.com/.../51057.what-s-being-deprecated-in-dynamics-365-version-9.aspx
Maybe in the new release April 2019?
Thank you in advance
""Deprecated" means we intend to remove the feature or capability from a future major release of Dynamics 365."
What I know, there is no word on when 10.0 will be released.
Business Applications communities


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I am a self-confessed Woolworths fan. And why not? A brand synonymous with quality and always on trend is definitely a front runner in South African chain stores. So with winter almost here, they always impress with their winter range. Here are a few items I'm saving up for because besides being the ultimate winter wardrobe essentials, they are oh so stylish.
Oh yes did I mention that you get a 10% WReward off on all women's boots? I suggest you calmly sprint to your nearest Woolworths and shop, shop, shop!


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If you've used a computer in the last 10 years (and let's face it, you probably have used one regularly) you have probably heard one or more of these terms and explanations:
- CPU. The Central Processing Unit is strictly defined as "the electronic circuitry within a computer that carries out the instructions of a computer program by performing the basic arithmetic, logical, control and input/output (I/O) operations specified by the instructions." They are some of the most important parts of a machine.
- Monitor. A graphics display of any kind. The complication with monitors comes when deciding on two things: resolution and technology. You might want a 4K capable monitor right now, or you only need a 720p one. Then you have to choose whether it is an LED, OLED or LCD. Or perhaps use a TV for a display.
- GPU. The Graphics Processing Unit. A dedicated video card that will help a computer push the limits of graphics be it in video games, rendering or editing video. The choices here are mainly between Nvidia and AMD cards, and each manufacturer has multiple tiers of GPUs to choose from. It can get complicated.
- PSU. The Power Supply Unit. You have to choose the correct wattage, as well as whether you want a modular PSU or not. This depends on the type of machine you are building, since more powerful computers need a lot more power than regular ones.
- Motherboard. The most important part of a PC. Everything we have in this list hooks up to the motherboard in one way or another. As such, it is very important to look for good quality MoBos, as they are popularly called. Aside from that, you need to be able to choose the correct type for socket for your CPU as well.
- RAM. Random Access Memory. The main choice here is with size: get 8, 16, 32 or even more Gigabytes of RAM? (The truth is that for most systems anything above 16 Gbs is overkill). But then there's the type of memory to get: DDR2, 3, 4 or 5, to name a few. It all comes down to how much you need to do and what you're willing to spend.
As you can see, this was only a small list designed to skim through the surface of some of these choices. For the rest of this article we will be centring on the CPU. To be more specific, we will centre on AMD CPUs' performance.
Let's be frank for a moment and simply state the fact that at the present time AMD's CPUs are simply not performing at the same level of Intel's CPUs. Sure, a CPU from both companies might advertise a clock speed of 2.5 Megahertz (MHz), but clock speed is not the be all and end all when it comes to a processor.
There are many components to a CPU:
- Logic Units
- Caches
- Registers
- Pathways
And a lot of other things. And all of them contribute to the performance of the CPU. These components are all incredibly complex, and the way they interact with each other, how they are designed to interact with each other will determine overall performance.
Both Intel and AMD use radically different cores. In AMD chips, every 2 of these cores share many of the same parts, which mean that they technically cannot be counted as 2 cores, but more like one-and-a-half cores. While this does not mean that programs that are well designed to take advantage of multiple cores don't get a benefit of these multi-cores, but it does limit the effectiveness of these multi-core processors in other situations.
There's also another fact that comes into play: processing stages. Different chips break down different tasks into manageable pieces, so a single process such as adding X to Y is broken down into a number of stages that the processor handles one at a time and then outputs the result. In most cases, the AMD processors take more cycles to complete the same operation or directive that an Intel processor would take. This means that even though both of these CPUs have the same clock speed, one of them is effectively taking twice the processing time in order to complete a task than the other.
There is a silver lining to all this, however: most of these issues will only affect your CPU's capacity to process whatever tasks you give it when it comes to extremely high performance use, and even then, it would just put more strain on the processor, but they will undoubtedly still handle it. When it comes to a regular user, the difference that these processors have in performance will not come into play.
On a final note, why not give SafeBytes DriverAssist a try? With this handy lightweight program, you get all the benefits of always having your drivers completely up to date, without the hassle of actually having to do it yourself!
Here are some of DriverAssist features:
- Automatic Driver Installation. You have no need to do anything else with DriverAssist other than install it onto your PC; the sophisticated software does all the work for you.
- Massive driver database always ensures you can identify hardware, even when Windows can't!
- Smart Driver ID. The DriverAssist software is based on sophisticated technology that automatically detects all of the appropriate PC drivers in a matter of seconds. It will then match them with the latest available version, and update every driver required by your system with a compatible version on a regular basis.
- Full driver backup and restore. DriverAssist helps you backup all of your data and restore your machine to the time when it was previously working if you encounter any problems.
- Enhanced USB management. DriverAssist also helps you manage USB devices. There is no need to worry about safely removing hardware with DriverAssist, as it makes the ejecting process and safely removing hardware from your PC extremely straightforward.
- 24/7 customer support. SafeBytes is on call 24/7 to assist you with any problems that you might encounter.


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HD Media
BRISTOL, Tenn. - Contura Energy Inc., which emerged from the bankruptcy of Alpha Natural Resources, announced on Wednesday a new metallurgical coal project in Logan County in its 2019 first quarter earnings report.
The company's board of directors approved investment in the "Lynn Branch Project," which it says has a high-quality, high volume met coal reserve of over 25 million tons.
"The project is expected to produce approximately 1 million to 1.2 million tons annually at its full estimated run rate, with a projected cash cost per ton in the low $60 range," according to the report. "The capital allocation required to bring the project online is estimated at approximately $25 million to $30 million and does not require an increase to previously guided capital expenditures."
The company said production is expected to commence in the second quarter of 2020.
Contura Energy reported its results for the first quarter of 2019 include the full quarter impact of the Alpha merger, which closed in November 2018.
"The Trading and Logistics segment sales in the first quarter of 2018 were largely driven by Alpha-related shipments, which are now included in the Central Appalachia Met (CAPP - Met) segment for the first quarter of this year," the report said.
Contura reported net income from continuing operations of $8 million, or $0.41 per diluted share, for the first quarter 2019. In the first quarter 2018, the company had net income from continuing operations of $58.3 million or $5.66 per diluted share.
Based in Tennessee, Contura operates the Cumberland mine in Waynesburg, Pennsylvania and several other Appalachian mines, as well assets in Wyoming.
About half of its coal revenue comes from metallurgical coal, which is used in steel making, according to data from the second half of last year.


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"Time isn't the main thing it's the only thing!"
Miles Davis
How can Red Dog VA help you?
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If you want to make more time to create, think, plan, learn, have fun or just to be, it may be time to consider outsourcing some of your work!
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