text:
The Grace Card
- Fecha al aire:
- Everything can change in an instant ... and take a lifetime to unravel. When Mac McDonald loses his son in an accident, years of bitterness and pain erodes his love for his family and leaves him angry with God ... and everyone else. Can Mac and his new patrol partner, Sgt. Sam Wright, somehow join forces to help one another when it's impossible to look past their differences--especially the most obvious one? Every day, we have the opportunity to rebuild relationships and heal wounds by extending and receiving God's grace. Offer THE GRACE CARD ... and never underestimate the power of God's love.
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Aylmer, Quebec
Aylmer | |
Sector within City of Gatineau | |
Rue Principale (Main Street)
Coordinates: 45°24′00″N 75°51′00″W / 45.40000°N 75.85000°WCoordinates: 45°24′00″N 75°51′00″W / 45.40000°N 75.85000°W | |
Country | Canada |
Province | Quebec |
Region | Outaouais |
RCM | Gatineau |
Incorporated | July 1, 1855 |
Merged | January 1, 2002 * |
Area | |
• Land | 88.86 km2 (34.31 sq mi) |
Population (2011) 2011 census |
• Total | 55,113 |
• Change ~2001 | +15.1% |
• Dwellings | 15,370 |
Demonym | Aylmerite |
Time zone | EST (UTC-5) |
• Summer (DST) | EST (UTC-4) |
Area code(s) | 819 |
Access Routes | Route 148 |
* Merged with Gatineau on January 1, 2002 |
Aylmer is a former city in Quebec, Canada. It became a sector of the City of Gatineau on January 1, 2002. Located on the Ottawa River and Route 148 it is a part of the National Capital Region. The population in 2011 was 55,113.[1] It is named after Lord Aylmer, who was a Governor General of British North America and a Lieutenant Governor of Lower Canada from 1830 to 1835.
It bills itself as the "Recreation Capital of the National Capital", given its many golf courses, green spaces, spas, marina and bicycle paths. There is little industry in the sector, the area being mainly residential. Virtually all the major shops, services and restaurants are located along Chemin d'Aylmer. The sector's indoor swimming pool and skateboard park are also located on that road.
Aylmer's population is about 30% anglophone ; 61% francophone[citation needed]and 9% other ; much of its workforce commutes across the river to Ottawa. As such it tends to be a very federalist area, with much of the population being bilingual.[citation needed]
Prior to its foundation, parts of Aylmer, like most surrounding areas of the Ottawa region, were often occupied as summer camps by the Algonquin First Nation population. The first European explorers known to reach the actual location of Aylmer were Nicolas-du-Vigneau and Samuel de Champlain during the early 17th century in their explorations west of Quebec City. It was only during the early 19th century that colonization began in the region; during the same period the foundations of the communities of Hull and Bytown were being established. In 1802, agricultural lots were given to Philemon Wright, an American from Massachusetts who was the first major developer/pioneer of the Ottawa Valley and Wright, in turn, gave most of his lots to his sons to operate as farms. Philemon Wright Jr. owned and operated the Chaudière Farm, established in 1816 at the end of the Britannia turnpike (more recently known as the Aylmer Road). After Philemon Jr's tragic death in 1821, Charles Symmes, who was Philemon Jr.'s first cousin, assumed the operations of the farm. Charles Symmes played a major role in the early development of Symmes' Landing, overseeing the construction of the Steamboat dock and the beautiful Inn, which still carries his name today. The post office and county registration office in Aylmer were opened in 1831; it was named after then Governor General Lord Aylmer. The village was first incorporated in 1847[2] and served as administrative centre for the region until 1897. A courthouse and jail that served the Outaouais region were built in 1852.[3] With the important shipbuilding yards on the banks of the Ottawa and its significant growth as one of the region's economic powerhouses of that time,[1] Aylmer was actually considered as a real contender to become the new Nation's Capital by Queen Victoria.[citation needed]
The Aylmer Boating Club was founded in 1890. The Club was renamed the Aylmer Yacht Club in 1900. In 1901, Moses Chamberlain Edey designed the clubhouse. By 1906, the Club was renamed the Victoria Yacht Club. In 1921, the Club burned down and was not rebuilt. [4]
During most of the 19th century, the town of Aylmer, like much of the Outaouais, was an important center for the wood industry. During that period several steam boats were built alongside the Deschênes Rapids and the Ottawa River across from Britannia. Railroad construction began during the early Canadian Confederation years. Meanwhile, the economy of Aylmer was more focused on the wood and wood pulp industries and much later, on tourism. In 1921, a destructive fire ravaged large sections of the village destroying dozens of homes and businesses. During the Great Depression Aylmer's biggest sawmill closed its doors. [2]
Aylmer would regain importance during the second half of the 20th century when, due to urban sprawling from the Ottawa and Gatineau areas, it became an important suburb to the region. In 1975 the villages of Lucerne and Deschênes, located just east of downtown Aylmer, were amalgamated. Several new residential developments were created on the northern and eastern side of old Aylmer. Numerous businesses and shopping malls were built along the Main Street including les Galeries Aylmer and the Glenwood Plaza, the latter being destroyed by a fire in 2005 and rebuilt. In addition, several golf courses, a Sheraton hotel, and a movie theater were added through the city. On August 4, 1994, a destructive tornado tore through the city damaging nearly 400 to 500 homes (including a dozen homes that were completely destroyed) and injuring at least 15 people. Damage figures were estimated at about $15 million. [3] Rated F3 on the Fujita scale, the tornado tracked for 8 kilometers and was one of the most intense tornadoes in history across the National Capital Region. [4] Aylmer also saw many power outages and very dangerous driving conditions throughout the North American ice storm of 1998, which left more than 4 million people without electricity, most of them in southern Quebec, western New Brunswick and Eastern Ontario, some of them for an entire month. The Canadian Forces were called in to assist with the relief efforts.
Before the amalgamation of the Urban Community Region of the Outaouais, Aylmer had a population exceeding 40,000 with additional growth after 2002 stemming from development in several areas of the sector, including the expansion of the Plateau de la Capitale neighbourhood which started in the former city of Hull in the early 1990s.
In recent years, efforts have been made to create the Boucher Forest protected area, in light of increasing suburban sprawl and housing development. The group that strives for protection of the Boucher Forest is the Fondation Forêt Boucher, the Boucher Forest Foundation. The forest is a reservoir of biodiversity in the region; at-risk and endangered species such as Panax quinquefolium (American ginseng), and Juglans cinerea (butternut) exist in this threatened ecosystem. The forest is bound by Highway 148 to the south, Chemin Vanier to the east, the Jardins Lavigne neighbourhood to the west (one of Aylmer's newer suburban areas), and Chemin Pink to the north. Furthermore, an old mining quarry is directly adjacent to the forest's northwest corner.
Roads and recreational pathways[edit]
Aylmer is served by provincial Route 148, known as the boulevard des Allumettières within the city of Gatineau, which extends from the Ontario border near Pembroke, to Montreal, about two hours away. Other main roads include the Chemin d'Aylmer/rue Principale, Lucerne Blvd., Vanier, Eardley, Broad/Klock, Wilfrid-Lavigne, Mountain Rd., Pink Rd. Aylmer is connected to Tunney's Pasture and Westboro in Ottawa by the Champlain Bridge, at the southeast corner.
It has been proposed to build an extension from Autoroute 50 in Gatineau that would come though Chelsea and central Aylmer to become a bridge between Deschênes and Brittania, but the plan is only tentative, mainly due to strong local opposition from the Brittania sector.
Aylmer is home to an effective and generally well-maintained network of bicycle paths that encircle the central portion of the area and run past many scenic locations, such as the Aylmer Marina and the Deschênes Rapids. The bike path system is maintained by the National Capital Commission.
Public transit[edit]
Public transit is provided by the Société de transport de l'Outaouais or STO, which runs twelve bus lines through the region (although many only operate during rush hour). The STO has been criticized by Aylmer residents—particularly youth—for not providing enough service to the area, and for not providing enough inter-sector bus lines (the eastern terminus for most Aylmer lines is in the Rideau Centre in downtown Ottawa). The STO is planning a bus rapid transit system known as Rapibus that would connect the Hull and Gatineau sectors, with the possibility of an expansion to Aylmer.
The railroad bedding still exists from Aylmer's now-defunct rail line, and pressure has been put on the STO to set up a light rail system in Gatineau that could connect to Ottawa's O-Train network via the Prince of Wales Bridge. If this were to happen, Aylmer could theoretically be served by light rail as well, but at present this appears highly unlikely.
In terms of population, Aylmer makes up about one third of the riding of Hull—Aylmer, which has elected a Liberal member of parliament in every federal election since its conception in 1984 — its predecessors, the ridings of Hull and Wright, in place from 1892 to 1984, also only ever elected Liberals. This, however, changed when the riding elected NDP MP Nycole Turmel to the House of Commons in the 2011 federal election.
In the 2005 Gatineau municipal election, Aylmer voters showed particularly strong support for current mayor Marc Bureau, over incumbent and former mayor of "old" Hull Yves Ducharme. Similar voting patterns appeared in the sectors of Buckingham and Masson-Angers, the other two "outlying" regions of Gatineau. This could be due to a perception among residents that the Ducharme administration was more focused on the urban core of the new city, as opposed to the periphery, as well as the rapid development of green-spaces into residential subdivisions. However, six months after the November election, residents are showing the highest level of dissatisfaction with the Bureau administration out of all the sectors, citing Aylmer's being left out of municipal processes, poor quality of municipal services, and little to no action to halt suburban sprawl. Interestingly, Masson-Angers and Buckingham residents are presently showing the most support for the new government.
Recently, the City of Gatineau administration had plans to reuse a former landfill site on Cook Road in the north end of the sector to build a new composting plant. A deal was planned with a non-profit organisation called La Ressourcerie to operate the site. However, local residents are strongly opposed with health and environmental concerns especially due to the past of the landfill site.[5] Despite displaying their fierce opposition, which included acts of intimidation and threats towards some councillors, at a Gatineau City Council Meeting, Mayor Marc Bureau mentioned that he will still build the plant at the Cook site, and that according to him it was the best possible site.[6] He later added that he will study other possibilities for the plant.
Aylmer's three wards are presently represented on the Gatineau city council by Stefan Psenak, André Laframboise and Alain Riel.
Amalgamation and de-Amalgamation[edit]
In 2002 the City of Aylmer became a part of Gatineau when the then-Parti Québécois government forcibly merged several clusters of cities and metropolitan areas throughout Québec. Residents of Aylmer were particularly against the amalgamation, citing fears of reduced municipal services, more suburban development, and a loss of cultural identity, as well as geographic differences (Hull and Gatineau arguably constitute a region of conurbation, whereas Aylmer was at the time separated by an expanse of sparsely inhabited green space).
A movement was started to halt the "forced fusion" of five cities surrounding Gatineau. The movement had particularly strong support in Aylmer. Signs reading "Je me souviendrai des fusions forcées" (literally, "I will remember forced fusions," a play on Québec's motto "Je me souviens") were a common sight.
When the Québec Liberal Party won the 2003 provincial election, the newly amalgamated former cities were given the opportunity to demerge. A referendum was held to decide the fate of the City of Gatineau which required a double vote: at least 35% of eligible voters from a given sector had to cast ballots, and more than 50% of these had to be in favour of de-amalgamation. Aylmer voters chose to separate from Gatineau but not enough ballots were cast, meaning Aylmer remained a sector of the larger city.
The voting outcome itself was done in such a way that even if Aylmer succeeded in de-amalgamating from Gatineau, they would only do so in name. The de-amalgamation claimed that once Aylmer was its own area once again, it would have to pay for all its own changes, while Gatineau would still be in control of the administration. This meant that the sector of Aylmer would receive no support from the administration that determined what was built where, but would still have to contribute 100% to the costs of the changes voted by the city of Gatineau.
Prior to the merger, Aylmer's residents and municipal laws had strongly opposed extensive construction programs. Following the amalgamation, many of the sector's prized green spaces were cut down for residential construction. Ex-Aylmer neighbourhoods like Wychwood and Village Lucerne have seen their undeveloped spaces sold to contractors. As a result of this unchecked development, there is a strong resentment of the current municipal administration among Aylmer Sector citizens.[citation needed]
List of pre-amalgamation mayors[edit]
List of mayors of Deschênes (1920-1974)[edit]
- John Routliffe (1920–1930)
- Israël Gravelle (1930–1937)
- Alexandre Madaire (1937–1944)
- Harry Rosenes (1944–1946)
- Jean Côté (1946–1951)
- Émilien Clairmont (1951–1953)
- Jean B. Côté (1953–1959)
- René Paulin (1959–1965)
- Jean-Paul Poirier (1965–1969)
- Kenneth Lloyd (1969–1973)
- Jean-Paul Poirier (1973–1974)
List of mayors of Lucerne (1879-1974)[edit]
Note:From 1879 to November 1964, this area was known as South Hull
English School Board[edit]
Western Quebec School Board
Provides English and French immersion education to primary and secondary students.
Primary schools[edit]
South Hull: Immersion primary school, located in Lakeview Terrace.
Rapides Deschênes: A francophone primary school, located on Vanier road, just south of Lakeview Terrace. Occupying three buildings, it teaches school from kindergarten to grade 6.The schools were originally known as St George ( grades 1 and 2 ), Notre Dame ( for girls, with a convent attached ) and St Medard ( for boys ). The schools had English and French classes in the 1950s and 1960s.
Vieux-Verger: A francophone primary school, located on Wilfrid-Lavigne boulevard. It teaches school from kindergarten to grade 6. Before the school was built, an orchard was present on that lot, which inspired the name for the school, which is French for "Old Orchard".
Euclide-Lanthier: A francophone primary school, located on Elizabeth street. It teaches kindergarten to grade 6.
Trois-Portages: A francophone primary school, located on Broad street, in the lot next to École Secondaire Grande-Rivière. It teaches kindergarten to grade 6.
École du Village:A francophone primary school divided into two separate buildings: Limoges and St-Paul. It teaches grades 1 through 6. Limoges (grade 1 to 3) is located on Symmes. St-Paul (grade 4 to 6) is located on Dalhousie.
Lord Aylmer School : Renamed after the amalgamation of Saint Mark's Elementary and Aylmer Elementary. The school has two campuses (located only across the street from each other). The former St. Mark's (known as the "Junior Campus") teaches students from kindergarten to grade 3. Aylmer Elementary (the "Senior Campus") teaches grades 4 to 6. Lord Aylmer Elementary has a program in English and French immersion. Located on Frank Robinson street.(Historical note: Aylmer Elementary was previously Aylmer High School, an English-language secondary school which was de-commissioned with the opening of the Philemon Wright High School in Hull in 1969.)
Montessori de l'Outaouais: Based on the Montessori theory of education, this French preschool and kindergarten is thirty years old. It is located at 161 Principale Street.
Secondary schools[edit]
École secondaire Grande-Rivière: a francophone high school, located on Broad street, this school teaches well over 2000 students, and supports an additional 100+ staff members, including teachers, administrators, janitors, and other service personnel. It is the largest secondary school in the sector. It sports a standard programme, a musical concentration programme, an artistic concentration programme, an IB Middle Years Programme, the International Programme (P.E.I), and several support programmes for students in difficulty. It also has a small community of highly active students who participate in the organization of school activities.
Other secondary schools are Symmes Junior High School (grades 7 and 8) and the recently constructed École secondaire D'Arcy McGee High School (grades 9, 10 and 11), both located on Blvd. du Plateau, this is because they are joined together.
Notable Aylmer individuals[edit]
- Meredith Henderson Gemini Award winning actress, now living in Los Angeles, CA
- Paul & Isabelle Duchesnay
- Polly Shannon Actress now living in Hollywood, CA.
- Dean DeBlois
- Merlin Bronques
- Charlie Major born in Aylmer
- Matthiew Klinck (Hollywood film director) born in Aylmer
- Larry Archambeault born in Aylmer, former hockey player
- Yannick Lupien Olympic swimmer (2000, 2004) silver medal World aquatic Montréal 2005
- Matthew Barnaby former NHL player
Geographic Location[edit]
Cantley Chelsea |
Pontiac | Hull Gatineau |
Ottawa River Bridge to: Ottawa, Ontario |
- Aylmer Nord
- Aylmer Park
- Champlain Park
- Connaught Gardens/Lucerne
- Des Cèdres
- Deschênes
- Glenwood/Vieux Moulin
- Jardins Lavigne
- La Seigneurie
- Lakeview Terrace
- Marina
- McCleod
- Mountain View
- Projet Pilon
- Queen's Park
- Rivermead
- Vieux Aylmer
- Wychwood
See also[edit]
- ^ Population calculated by combining Census Tracts 5050822.03, 5050822.04, 5050821.02, 5050821.01, 5050800.00, 5050820.00, 5050811.02, 5050811.01, 5050810.02, 5050810.01
- ^ "Aylmer, Quebec". The Canadian Encyclopedia.
- ^ "Heritage". City of Gatineau.
- ^ Moses Chamberlain Edey Dictionary of Architects in Canada http://dictionaryofarchitectsincanada.org/architects/view/1549
- ^ Residents raise stink over Gatineau composting plant
- ^ Duquette, Patrick, Gatineau fera son compostage dans l'ancien depotoir Cook, (Gatineau will do its compositing plant at former Cook landfill site), Le Droit, Ottawa, September 20, 2006, page 3.
- ^ Aylmer 1847-1974.PDF
- ^ http://www.ville.gatineau.qc.ca/Archives/Aylmer%201975-2001_Conseil.pdf
- ^ "Municipalité de Deschênes – Liste des maires et des conseillers". City of Gatineau.
- ^ "http://craoutaouais.ca/repertoire/Web/hull-sud.html". Répertoire des élus municipaux de l'Outaouais de 1845 à 1975 (in French). Archives nationales du Québec.
- ^ "Municipalité de Lucerne - Liste des maires et des conseillers". City of Gatineau.
External links[edit]
Wikimedia Commons has media related to Aylmer, Quebec. |

subset: commoncrawl


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A bit about me:
I spend much of my time at North Shore University Hospital in Manhasset, NY, volunteering. My children convinced my husband to buy me a Macintosh PPC for Mother's Day in 1994. Before that, I had never used a computer I really could "play" with--but I sure have learned how to play since! I do a fare amount of surfing, through CompuServe.
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Destination: Hawaiian Islands Article Source: Outrigger
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"Celebrate Kani Ka Pila" will offer an unforgettable concert from some of Hawaii's most noted musical artists, not to mention a grazing menu of delectable island cuisine. It addition to supporting our wonderful island musicians, the concert will also benefit MusiCares, an organization that provides support to musicians and other industry people who may need a little help getting back on their feet.
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$ 600 Standard work on Chinese painting" Osvald Sirén, Chinese Painting: Leading masters and principles, vols. I to VII, facsimile (orange cloth), Hacker Art Books Inc., New York 1973. 11 1/8 x 8 ½ inches. Vol. I Early Chinese painting Vol. II The first millennium. The Sung period Vol. III The first millennium. Plates Vol. IV The later centuries. The Yüan and early Ming masters Vol. V The later centuries. The later Ming and leading Ching masters Vol. VI The later centuries. Plates Vol. VII Annotated lists of paintings and reproductions of painting by Chinese artists Covers with some rubbing and slightly soiled on the cut of the paper, inside excellent condition, binding overall almost as good as new. |
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Fort St. Philip
Fort St. Philip
Fort St. Philip in the U.S. Civil War
Location | Plaquemines Parish, Louisiana, US |
Nearest city | Triumph, Louisiana |
Built | 1746 |
Governing body | Private |
NRHP Reference # | 66000380 |
Significant dates | |
Added to NRHP | October 15, 1966[1] |
Designated NHL | December 19, 1960[2] |
Fort St. Philip is an historic masonry fort located on the eastern bank of the Mississippi River, about 40 miles (64 km) upriver from its mouth in Plaquemines Parish, Louisiana. It formerly served as military protection of New Orleans, Louisiana, some 80 miles (130 km) up the river, and of the lower Mississippi River.
The first fort on this location, "San Felipe," was constructed in the 18th century during the period of Spanish control of Louisiana.
The fort served a role in protecting the United States from the British invasion during the War of 1812, seeing ten days of battle in January 1815, the 9th to the 18th, inclusive. Specifically, the fort held its defenses against British Navy vessels who were bombarding it, in a final attempt to invade Louisiana following the defeat of the British Army near New Orleans.
The current fort was constructed, along with Fort Jackson on the river's western bank, as a coastal defense for New Orleans and the Mississippi, upon the urging of Andrew Jackson. It was the site of a twelve-day siege in April 1862 by Union forces during the American Civil War.
It was declared a National Historic Landmark in 1960.[2][3]
In the 1930s the fort was used as a tanning factory. This explains the old railroad tracks and fire hydrants.[citation needed]
From 1978 through 1989 the fort complex served as the site of an intentional, nonsectarian spiritual community called Velaashby,[4][5] named by conjoining the surnames the original and subsequent private property owners respectively. The community members numbered as many as 16 at any one time and were known as the Christos family. They lived in four buildings—three two-story officers quarters and an officers club—that remained from the re-fortification of the site during the 1898 Spanish–American War.[2][5]
Fort St. Philip remains privately owned and in a state of bad deterioration. It was heavily damaged in hurricanes Katrina and Rita. According to the National Park Service, the owner reported that only the original brick fort and the concrete structures from the time of the Spanish–American War remain.[2]
The site is accessible only by boat or helicopter, and following erosion of the small levee is now subject to flooding during high water levels of the Mississippi River.[2]
- ^ "National Register Information System". National Register of Historic Places. National Park Service. 2007-01-23.
- ^ a b c d e "Fort St. Philip". National Historic Landmark summary listing. National Park Service. Retrieved 2008-02-01.
- ^ Patricia Heintzelman (1978). "National Register of Historic Places Inventory-Nomination: Fort St. Philip". National Park Service.32 KB and Accompanying 6 photos, aerial photos from 1935 and others, undated. PDF (3.09 MB)
- ^ Sutton, James (2007-09-10). "Ft. St. Philip -- Vella-Ashby". Panoramio. Retrieved 2008-11-08.
- ^ a b Raber, Rick (1984-02-26). "The Christos Experiment". Times-Picayune, Dixie supplement (New Orleans).
External links[edit]
Wikimedia Commons has media related to Fort St. Philip, Louisiana. |
- First Siege of Fort St. Philip (1815)—eyewitness accounts, as published in the Louisiana Historical Quarterly.
- Second Siege of Fort St. Philip (1862)—Chapter 15 of Kendall's History of New Orleans
Coordinates: 29°21′50″N 89°27′46″W / 29.36389°N 89.46278°W
- American Civil War forts
- Forts in Louisiana
- Louisiana in the American Civil War
- National Historic Landmarks in Louisiana
- Buildings and structures in Plaquemines Parish, Louisiana
- Ruins in the United States
- Spanish forts in the United States
- Colonial forts in Louisiana
- Forts on the National Register of Historic Places in Louisiana


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I have about 4 of these full size ionic breeze around the house and bout 4 of the bathroom size plugins. I read all about these but it is allways mixed results. I read Yes they work they create an ozone byproduct and nothing knocks out smell like Ozone. Then I hear not to use them near your plants because they create ozone it could be bad for you and your plants but they do cut the smell down a small %.... So I want to ask has anyone here used these with good success no damage to your plants ect ect.....
they can be harmful to your plants nut in larger dose than a few can put out. we have 3 and love them.
I haven't read any educated posts citing any evidence that these units are bad for your plants, but I've read plenty of threads stating that they do not effectively counter the odor of flowering plants. I have two relatively new Ionic Breeze units in my home, but I will not be using them for odor control....but I have the ability to vent to the outdoors. I guess the decision on whether or not to use them depends on your intake/exhaust options. If I was running an enclosed grow I might give it a try but, from what I've read, I honestly wouldn't set my hopes very high.
I didn't think the ionic breeze created ozone, and I don't think ionization alone would be effective. If you have them test them. If they do generate ozone it would not damage plants, but don't put them right in the grow area, put them in the room the grow area is in.
also dont know if the ionic breeze produces ozone.I have an ozone generator and use it in the room I vent into.my mentor (who gave me the ozone gen)warned me not to put it w/ the plants as it would take the smell away from the plants and affect the taste.
Well I put one in the closet with my 1 plant and the closet doesnt smell at all anymore after just 24 hours. So I am thinking about adding another one in the closet when flowering. I will see how the one Ionic works first . I hope to start flowering in 1-3 weeks so I will try and post on how they work in flowering.
i tried those and they did not cover my smell while flowering. i had to get carbon filter. but i had more than one plant
Air ionizers are not ozone generators. Those Ionic Breeze devices are actually designed to release as little ozone as possible. O3 is considered a pollutant and those air purifiers are only supposed to produce a certain amount as per federal guidelines. I had the largest Ionic Breeze they make and tried using it to remove smoke, and the smell of an oz of stinky ass Kush in a zip lock bag (this was back in the day). Even on high with the "ion boost" or whatever it's called on, it took a while for the thing to have any effect. It did eventually help, but not by much. I If you already have it, use it. Otherwise just buy an ozone generator/carbon filter or just open some windows
Yeah, because having your neighbors smelling the strong aroma of mary jane would be great. Do feds come on this site and post bullshit to get people busted or what?
Wow, smug much? I honestly never thought I'd see this kind of behavior on an MJ board. Are you suffering through a dry spell or something? You just took my suggestion out of context and assumed it applies literally to anyone in any situation. If I'm a narc, then you're a journalist. Think about it dude. Someone with a bunch of plants would realize that opening the window to get rid of the smell is a bad idea. If OP has a whole op going (lol op op), he's not going to blow up his spot just because he reads "or just open some windows " at the end of some post on an internet forum. The sentence ends with freakin mr. green blazing a J, how do take it that seriously? Ironically, OP is talking about one plant in a closet. I think it's safe to say that you wouldn't be able to smell it unless you're like 5ft from the window. Yes, he could have put a fan in the window and a decent amount of air actually leaves the room. Hell, he might even be one of those people with a fetish for opening and closing doors, or he may live in an urban area with strict penalties for growing a plant, and it's also possible that he told someone about the plant and cops may now be watching his property. You'd have to be an idiot to open windows to vent grow odor unless you know that it's safe. It seems that you are also assuming OP is retarded. I sincerely hope you understand what I was saying now. In the future, I will try to accompany any controversial statement with an asterisk and a footnote to dissuade the trolls and aid people that just don't get it.


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Please use this identifier to cite or link to this item: http://hdl.handle.net/1811/5381
Files | Size | Format | View |
V68N02_123.pdf | 347.5Kb |
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Title: | Heats of Hydration and Absolute Heats of Formation of Aqueous Ions |
Creators: | Myers, R. Thomas |
Issue Date: | 1968-03 |
Citation: | The Ohio Journal of Science. v68 n2 (March, 1968), 123-127 |
Abstract: | Accurate calculations are made for the hydration energies of the alkali halides, using the most recent data. These results, and the average of theoretical calculations made by others for the individual ions, are used to determine the most probable hydration energy of the individual ions. These in turn are the basis for calculating the absolute enthalpy of formation of aqueous H+: 98.6 kcal/g-equiv. A Born-Haber cycle can then be used to calculate the hydration energy of any other ion for which the enthalpy of the gaseous ion is known. |
Description: | Author Institution: Department of Chemistry, Kent State University, Kent, Ohio 44240 |
URI: | http://hdl.handle.net/1811/5381 |
Rights: | Reproduction of articles for non-commercial educational or research use granted without request if credit to The Ohio State University and The Ohio Academy of Science is given. |
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US Offers $10m For Pakistani Militant?
The United States has announced a $10 million reward for information leading to the capture of Hafiz Mohammad Saeed, a Pakistani militant leader accused of orchestrating the 2008 attacks in Mumbai, India, who in recent months has emerged at the vanguard of a prominent anti-US political-lobbying interest.
Wendy Sherman, the US undersecretary of state for political affairs, announced the reward while in India on Monday for help finding Saeed, described as the leader of the militant group Lashkar-e-Taiba. She also announced $2 million for information leading to the capture of Hafiz Abdul Rahman Makki, Saeed's brother-in-law.
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Page 1
Apoptotic surge of potassium currents is mediated
by p38 phosphorylation of Kv2.1
Patrick T. Redman*, Kai He*, Karen A. Hartnett*, Bahiyya S. Jefferson*, Linda Hu†, Paul A. Rosenberg†,
Edwin S. Levitan‡, and Elias Aizenman*§
Departments of *Neurobiology and‡Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261; and†Department of Neurology
and Program in Neuroscience, Children's Hospital and Harvard Medical School, Boston, MA 02115
Edited by Michael V. L. Bennett, Albert Einstein College of Medicine, Bronx, NY, and approved January 4, 2007 (received for review November 15, 2006)
Kv2.1, the primary delayed rectifying potassium channel in neu-
have described Kv2.1 phosphorylation events affecting channel
gating and the impact of this process on cellular excitability. Kv2.1,
however, also provides the critical exit route for potassium ions
during neuronal apoptosis via p38 MAPK-dependent membrane
insertion, resulting in a pronounced enhancement of K?currents.
mutants identify a p38 phosphorylation site at Ser-800 (S800) that
death. In addition, a phospho-specific antibody for S800 detects a
p38-dependent increase in Kv2.1 phosphorylation in apoptotic
neurons and reveals phosphorylation of S800 in immunopurified
channels incubated with active p38. Consequently, phosphoryla-
tion of Kv2.1 residue S800 by p38 leads to trafficking and mem-
brane insertion during apoptosis, and remarkably, the absence of
death program.
apoptosis ? ion channel ? MAPK
Blocking K?channels, or increasing the extracellular K?con-
centration, effectively attenuate cell death in many apoptotic
models (2–6), including oxidant exposure in cortical and mid-
brain dopaminergic neurons (7–10). With the use of dominant
negative mutant subunits, Kv2.1, the major component of the
delayed rectifier K?current in neurons (11, 12), was identified
as the channel responsible for mediating the apoptotic K?
current surge in cortical neurons (13). During oxidant-induced
neuronal apoptosis, the liberation of intracellular Zn2?from
metal binding proteins (7) leads to the activation of p38 MAPK,
which precedes and is necessary for the characteristic K?current
surge (8, 9). This enhancement of K?currents is caused by a
soluble N-ethylmaleimide-sensitive factor attachment protein
receptor-dependent insertion of new Kv2.1-encoded channels,
rather than an alteration in the properties of existing surface
channels, such as a change in activation kinetics (3, 14). Despite
this information, the molecular process connecting p38 activa-
tion and the apoptotic membrane insertion of Kv2.1 K?channels
had heretofore remained undefined.
Here, we combine several experimental approaches to estab-
lish a direct link between active p38 and Kv2.1 during apoptosis
after a sequence-based prediction model (15) was used to
identify a putative phosphorylation site for the MAPK on the C
terminal of the channel. First, alanine substitution of Ser-800
(S800) in Kv2.1 completely abolished the apoptotic enhance-
ment of K?currents. Second, a cysteine-containing mutant of
Kv2.1 and a thiol-reactive covalent inhibitor were used to
demonstrate that S800 is critical for membrane insertion of the
channel during apoptosis. Third, expression of phospho-mimetic
mutant channels resulted in significant increases in basal K?
current densities. Fourth, a phospho-specific antibody directed
at S800 detected a p38-dependent increase in phospho-Kv2.1
nhancement of voltage-gated K?channel activity, resulting
in K?efflux, is an essential step in neuronal apoptosis (1).
levels in apoptotic neurons and revealed phosphorylation of
immunopurified Kv2.1, but not of Kv2.1(S800A), incubated with
active p38. Most significantly, Kv2.1(S800A) did not support
apoptosis mediated by the WT channel in a recombinant ex-
pression system. This study establishes that p38-mediated phos-
phorylation of Kv2.1 is necessary and sufficient for its apoptotic
trafficking and completion of the cell death program.
S800 in Kv2.1 Is Necessary for the Apoptotic Surge in K?Currents.
Scansite (http://scansite.mit.edu), a program that predicts pro-
tein phosphorylation sites based on proteomic and biochemical
data (15), was used to search for potential MAPK targets in
Kv2.1 (GenBank accession no. NM?013186). A medium-
stringency scan revealed a single serine residue at position 800
(Fig. 1A) as a potential phosphorylation target for both p38 and
ERK. However, the Scansite score for this sequence suggested
a substantially better match for p38 than for ERK. S800 resides
in the serine- and threonine-rich intracellular C-terminal tail of
the protein (Fig. 1A), where other phosphorylation sites exist
(16–18). We thus examined the role of S800 in the up-regulation
of K?currents during oxidant-induced apoptosis in a recombi-
nant expression system.
Whole-cell electrophysiological recordings were performed
on CHO cells transiently expressing either WT Kv2.1 channels
or a nonphosphorylatable mutant, Kv2.1(S800A). CHO cells
have no endogenous voltage-gated K?channels (19), but can be
induced to readily undergo oxidant-induced apoptosis after
expressing Kv2.1 (13), and, like neurons, show a pronounced K?
current enhancement during this process (14). Recordings were
obtained under control conditions and after treatment with the
oxidant apoptogen 2,2?-dithiodipyridine (DTDP; refs. 7 and 14).
Electrophysiological measurements were routinely performed
3 h after oxidant exposure, a time when a robust K?current
surge is well established (8, 13, 14). As shown (14), currents
mediated by WT Kv2.1-encoded channels were substantially
enhanced after the apoptotic stimulus (Fig. 1B Left). In contrast,
no current surge was observed in CHO cells expressing
Kv2.1(S800A) after the oxidant treatment (Fig. 1B Right), indi-
cating that S800, a putative target for p38-mediated phosphor-
ylation, is required for the increase in K?currents during
Author contributions: P.T.R. and K.H. contributed equally to this work; P.T.R., K.H., P.A.R.,
E.S.L., and E.A. designed research; P.T.R., K.H., K.A.H., and B.S.J. performed research; L.H.
and P.A.R. contributed new reagents/analytic tools; P.T.R., K.H., K.A.H., B.S.J., and E.A.
analyzed data; and P.T.R., P.A.R., E.S.L., and E.A. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS direct submission.
§To whom correspondence should be addressed. E-mail: firstname.lastname@example.com.
This article contains supporting information online at www.pnas.org/cgi/content/full/
© 2007 by The National Academy of Sciences of the USA
3568–3573 ?
February 27, 2007 ?
vol. 104 ?
no. 9www.pnas.org?cgi?doi?10.1073?pnas.0610159104
Page 2
The apoptotic surge in K?current observed in both cortical
neurons and CHO cells is the result of trafficking and soluble
N-ethylmaleimide-sensitive factor attachment protein receptor-
dependent exocytotic insertion of new Kv2.1 channels into the
plasma membrane (14). New channel membrane insertion was
detected by using the cysteine-containing K?channel mutant
Kv2.1(I379C) (20). The presence of this mutation allows the
thiol reagent (2-trimethylammoniumethyl) methanethiosulfate
(MTSET) (21, 22). Virtually all previously existing surface
Kv2.1(I379C) channels can therefore be silenced after a 5- to
10-min exposure to MTSET. With existing plasma membrane
Kv2.1 channels permanently blocked, the insertion of new
channels is easily detected electrophysiologically by the appear-
ance of new currents under MTSET-free conditions (14). This
assay, which has the distinct advantage of detecting the insertion
of new functional channels, has been validated by parallel
biotinylation studies (14).
To investigate the role of S800 in the apoptotic insertion of
new channels, the double mutant Kv2.1(I379C, S800A) was
generated. Whole-cell recordings first confirmed the preserva-
tion of functional silencing by MTSET in this double mutant
channel (Fig. 2A). Next, CHO cells expressing either
Kv2.1(I379C) or Kv2.1(I379C, S800A) were exposed to MTSET
for 10 min to block all existing surface channels. The cells were
then exposed to oxidant to trigger the apoptotic cascade. Elec-
trophysiological recordings 3 h later revealed the presence of K?
currents in Kv2.1(I379C)-expressing cells (Fig. 2B), similar to
what was reported earlier (14). However, little or no current
could be measured in CHO cells expressing Kv2.1(I379C,
S800A), indicating a lack of apoptotic membrane insertion of
these mutant channels (Fig. 2B). This result not only accounts
for the lack of Kv2.1(S800A)-mediated apoptotic current surge
observed earlier, but demonstrates that phosphorylation of S800
is necessary for Kv2.1 membrane insertion during the cell death
Negatively Charged Amino Acids at Position 800 Mimic the Apoptotic
Surge. Recordings were performed from CHO cells transiently
expressing two additional variants of Kv2.1 to test whether
substitution of S800 with negatively charged residues would
mimic the apoptotic increase in currents observed in WT
channels. Indeed, basal current densities in untreated
Kv2.1(S800D) and Kv2.1(S800E) mutant-expressing cells dis-
played significantly enhanced amplitudes, comparable to those
observed in DTDP-treated Kv2.1-expressing cells (Fig. 3). Ox-
idant treatment of cells expressing these mutations did not
produce any additional current enhancement (data not shown).
Rather, a slight decrease in the currents was observed under
these conditions, likely the result of cellular damage by the
injurious stimulus. These results further support the role of S800
phosphorylation as a key mediator of the K?current surge
during apoptosis.
Fig. 1.
site on the cytoplasmic C terminus. (B) (Upper Left) Representative whole-cell
treatment conditions. Currents were obtained 48 h posttransfection and
densities from Kv2.1-expressing CHO cells under control (n ? 25) and DTDP
(n ? 12) treatment conditions (*, P ? 0.05; t test). Currents were induced with
a voltage step to ?5 mV from a holding potential of ?70 mV and normalized
to cell capacitance. (Upper Right) Representative whole-cell K?currents from
Kv2.1(S800A)-expressing CHO cells recorded under control and DTDP treat-
ment conditions. (Lower Right) Mean ? SEM. K?current densities from
Kv2.1(S800A)-expressing CHO cells recorded under control (n ? 24) and DTDP
(n ? 24) treatment conditions.
Ser-800 of Kv2.1 is critical for apoptosis-associated K?current en-
Fig. 2.
Kv2.1. (A) MTSET blocks Kv2.1(I379C) channel containing the Kv2.1(S800A)
mutation. Calibration was 2 nA, 10 ms. (B) (Upper) Diagram representing the
experimental protocol designed to measure the appearance of new K?chan-
nels during the apoptotic process. Shown are representative whole-cell K?
currents obtained from Kv2.1(I379C)- and Kv2.1(I379C, S800A)- expressing
of ?70 mV. Calibration was 1 nA, 10 ms. (Lower) Mean ? SEM. K?current
densities recorded from Kv2.1(I379C)-expressing CHO cells (n ? 17) and
Kv2.1(I379C, S800A)-expressing CHO cells (n ? 20) 3 h after the sequential
MTSET/DTDP treatments (*, P ? 0.01; t test). These currents represent the
newly inserted channels during apoptosis.
S800A mutation blocks apoptogen-induced membrane insertion of
Redman et al.
February 27, 2007 ?
vol. 104 ?
no. 9 ?
Page 3
S800 Phosphorylation Is p38-Dependent in Neurons Undergoing Apo-
ptosis. Biochemical studies were performed to detect phospho-
S800 levels in primary cortical neurons after oxidative injury.
Cell lysates were subjected to gel electrophoresis, and the
resulting blots were probed with a commercially available anti-
body directed against amino acids 841–857 of the C terminus of
Kv2.1 (Kv2.1; Alomone Labs, Jerusalem, Israel) or with an
antibody that we generated against the phosphorylated form of
S800 [pKv2.1; supporting information (SI) Fig. 6]. Immunoblots
probed with Kv2.1-directed antibodies revealed the presence of
two major bands in cortical extracts: a diffuse band near 100 kDa
and another, more compact band, at ?80 kDa (SI Fig. 7; see also
refs. 23–25). The diffuse band near 100 kDa likely represents
multiple, non-p38 dependent phosphorylation states of the
predicted full-length channel (18, 26), whereas the 80-kDa band
may reflect the presence of a protein generated by an alternative
mRNA isoform (23, 27) or a proteolytic fragment. Importantly,
preincubation of the two antibodies with their respective immu-
nizing peptides completely blocked both immunoreactive signals
(SI Fig. 7).
Phospho-S800 levels in cortical neuronal lysates were assessed
by using the pKv2.1 antibody under control conditions and after
oxidative injury. We observed a significant increase in pKv2.1
immunoreactivity in cortical neurons undergoing apoptosis,
compared with vehicle-treated controls (Fig. 4 A and B). This
increase was particularly apparent in the compact, 80-kDa band.
Importantly, SB-239063, a selective p38 inhibitor (8, 28), pre-
vented oxidant-mediated increases in pKv2.1 immunoreactivity
(Fig. 4 A and B). These data confirm a p38-dependent increase
in phospho-S800 levels in endogenous Kv2.1 channels from
cortical neurons undergoing apoptosis.
Active p38 Phosphorylates S800 in Cell-Free Assays. To determine
whether p38 could phosphorylate Kv2.1, we performed a cell-
free assay in which recombinantly expressed, immunoprecipi-
tated poly myc-tagged WT Kv2.1 (myc-Kv2.1; refs. 14 and 29)
was used as a substrate for purified, active p38. We observed that
myc-Kv2.1 protein exposed to active p38 produced a phospho-
specific immunoreactive band (Fig. 4C). Similar experiments
performed with an S800A mutant of the myc-tagged Kv2.1
yielded no phospho-specific signal (Fig. 4C). These results
indicate that active p38 can directly induce selective phosphor-
ylation of residue S800 of Kv2.1.
S800A Mutation Disrupts Kv2.1-Mediated Apoptosis. Unlike trun-
cated or pore mutants of the channel (13), Kv2.1(S800A) does
not function as a dominant interfering form in neurons under-
going apoptosis, but simply enhances the overall basal current
amplitude without eliminating the contribution of endogenous
channels (data not shown). As such, the requirement for S800
phosphorylation in apoptosis was evaluated in Kv2.1-expressing
CHO cells under oxidant exposure conditions that are normally
sublethal to vector-expressing control cells (13). A 15-min
exposure to 30 ?M DTDP was sufficient to induce apoptosis in
?50% of WT Kv2.1-expressing cells (Fig. 5). However, the same
oxidative insult was completely innocuous to CHO cells express-
ing Kv2.1(S800A) mutant channels. As a negative control, we
assayed the viability of CHO cells expressing a nonconducting,
double-pore mutant Kv2.1(W365C, Y380T) channel (12). As in
cells expressing Kv2.1(S800A), the oxidant was not toxic to cells
expressing the double-pore mutant channels. These data dem-
onstrate the requirement for functional Kv2.1 channels contain-
ing the phosphorylatable amino acid residue S800 to complete
the apoptotic program.
The results presented in this study reveal an important and
previously unrecognized mechanism of Kv2.1 regulation that has
a critical impact on apoptosis. Kv2.1 is subject to extensive
phosphorylation and dephosphorylation reactions, which alter
Fig. 3.
from Kv2.1-expressing CHO cells (Left), Kv2.1(S800D)-expressing CHO cells
(Center), and Kv2.1(S800E)expressing CHO cells (Right). Currents were ob-
?35 mV from a holding potential of ?70 mV. Calibration was 5 nA, 15 ms.
(Lower) Mean ? SEM. Current densities from Kv2.1-expressing CHO cells (n ?
13) (Left), Kv2.1(S800D)-expressing CHO cells (n ? 17) (Center), and
Kv2.1(S800E)-expressing CHO cells (n ? 14) (Right) (*, P ? 0.05; t test).
Substitution of S800 with negatively charged amino acids mimics
Fig. 4.
of Kv2.1-S800 is regulated by p38 MAPK in cultured cortical neurons. Cortical
cultures were pretreated with either vehicle or the specific p38 MAPK inhib-
itor SB293063 (SB; 20 ?M for 10 min) and then incubated with either vehicle
or DTDP (100 ?M for 10 min). Immunoblots were probed with anti-pKv2.1
antibody (1:5,000; Top), a commercially available anti-Kv2.1 antibody
(Alomone Labs; 1:1,000; Middle), and an anti-GAPDH antibody (Novus Bio-
logicals, Littleton, CO; 1:5,000; Bottom) to confirm equal protein loading
between lanes. These data are representative of results from three indepen-
dent experiments. (B) Oxidative injury significantly increases S800 phosphor-
ylation in cortical neurons in vitro. Optical density measurements of the
the 80-kDa band were quantified by using Scion Image software. Values
represent the mean ? SEM normalized pKv2.1 signal (to Kv2.1 immunoreac-
tivity) of both bands as a percentage of vehicle-treated control from three
separate, independent experiments (*, P ? 0.01; ANOVA/Dunnett). Note that
the observed increase in S800 phosphorylation was effectively blocked with a
p38 MAPK inhibitor. (C) S800 of Kv2.1 is phosphorylated by p38? in a cell-free
mM MgCl and 50 ?M ATP), and 50 ?g of activated p38? kinase. Reactions
containing kinase buffer and Mg/ATP and kinase buffer alone were used as
controls. Immunoblots (IB) were probed with the pKv2.1 antibody (1:12,000)
and the Kv2.1 antibody (1:3,000; Alomone Labs).
Phosphorylation of Ku2.1 by p38. (A) DTDP-induced phosphorylation
3570 ?
www.pnas.org?cgi?doi?10.1073?pnas.0610159104Redman et al.
Page 4
the functional properties of this channel by influencing gating,
and thereby, cell firing properties (18, 30). We have found,
however, a phosphorylation target for p38 on the C-terminal of
Kv2.1 that is vital for the apoptotic surge of K?currents
observed in apoptosis and the completion of the cell death
program. Interestingly, a recent report by Park et al. (18)
reported that S800 is a phosphorylation target, albeit by a
then-unidentified kinase. They further showed that S800 was not
one of several C-terminal sites subject to calcineurin-dependent
dephosphorylation and did not participate in the graded regu-
lation of Kv2.1 gating. We have found here that S800 is a unique
p38 phosphorylation target on Kv2.1 that is required for channel
trafficking and cell death.
Phosphorylation of S800 is not necessary, in and of itself, for
normal Kv2.1 trafficking and plasma membrane insertion, as
Kv2.1(S800A) mutant channels can be functionally expressed.
Indeed, normal neurons overexpressing this construct simply
have larger K?currents, which explains why this mutated form
of Kv2.1 cannot function as a dominant interfering channel
during neuronal apoptosis. Nonetheless, our studies in CHO
cells reveal that under injurious situations this site is critical for
de novo channel insertion, resulting in the current surge neces-
sary for cytoplasmic K?loss and cell death.
Earlier studies revealed that the Kv2.1-mediated current surge
during apoptosis is a result of the insertion of new channels into
the plasma membrane (14), and the p38 MAPK signaling
pathway is instrumental in this process (8, 9, 31). Interestingly,
p38 activation by TNF? has also been implicated in the poten-
tiation of tetrodotoxin-resistant Na?currents in sensory neu-
not been documented (32). The MAPK can phosphorylate
Nav1.6-encoded Na?channels, but in this case the resulting
interaction produces a decrease, rather than an increase, in Na?
currents (33). In addition, another member of the MAPK family,
ERK1/2, phosphorylates Kv4.2, a potassium channel responsible
for A-type K?currents in neurons (34, 35), resulting in a
down-regulation of dendritic K?currents in CA1 hippocampal
pyramidal neurons (36). Collectively, this information indicates
that MAPKs participate in cell signaling events that alter the
contribution of voltage-gated ion channels to overall neuronal
Kv2.1 is required for an entirely different, but critical process:
completion of the apoptotic program.
Several issues remain to be addressed. Primarily among them,
the molecular mechanism linking p38-induced phosphorylation
of Kv2.1 to the membrane insertion of this channel remains to
be determined. We previously observed that cleavage and
inactivation of the exocytotic soluble N-ethylmaleimide-sensitive
factor attachment protein receptor proteins SNAP-25 and syn-
taxin was sufficient to prevent the apoptotic K?current surge
(14). Importantly, both of these proteins are known to directly
interact with Kv2.1 (37): SNAP-25 with the cytoplasmic N-
terminal (38), and syntaxin with two domains of the cytoplasmic
C-terminal tail (39). Overexpression of syntaxin and Kv2.1
inhibits channel surfacing and leads to a decrease in Kv2.1-
mediated current density (39). It is thus tempting to speculate
that p38 phosphorylation of Kv2.1 leads to changes in its
association with syntaxin or other proteins, promoting, in turn,
exocytosis of channel containing vesicles. In addition, Kv2.1
subunits consistently colocalize with cortical neuron subsurface
cisterns, compressed stacks of smooth endoplasmic reticulum
situated ?5–8 nm beneath the plasma membrane (40). Thus, the
possibility exists for cisternae to serve as a nearby holding area
for channels destined for the plasma membrane (41), with p38
phosphorylation serving as the molecular switch necessary to
free Kv2.1 from cisternal retention during apoptosis.
In summary, we have identified a putative p38 phosphoryla-
tion target residue on the C-terminal of Kv2.1. Importantly,
mutation of S800 to a nonphosphorylatable residue is sufficient
to disrupt the apoptotic cascade initiated by oxidative injury and
block cell death. The work described here illustrates a critical
mechanistic link between oxidant-induced Zn2?release (7), p38
MAPK activation (8, 9), the ensuing Kv2.1-mediated apoptotic
K?current surge (13, 14), and cell death. Thus, prevention of
Kv2.1 phosphorylation by p38 is a potential novel therapeutic
stress conditions.
Materials and Methods
Plasmids and Site-Directed Mutagenesis. The mammalian expres-
sion vector encoding WT Kv2.1 was the gift of J. Trimmer
(University of California, Davis, CA). The polymyc-tagged
Kv2.1 plasmid was from K. Takimoto (University of Pittsburgh,
Pittsburgh, PA). The FLAG-tagged dominant negative
Kv2.1(W365, Y380T) plasmid was provided by J. Nerbonne
(Washington University, St. Louis, MO). S. Korn (National
Institutes of Health, Bethesda, MD) provided the Kv2.1(I379C)
vector. Mutagenesis of these cDNAs was performed with a
QuikChange XL kit (Stratagene, La Jolla, CA) according to the
manufacturer's directions. Primers containing the desired mu-
tations (S800A, S800D, S800E) were obtained from Integrated
DNA Technologies (Coralville, IA). Mutations were confirmed
by sequencing. A plasmid encoding EGFP (pCMVIE-eGFP;
Clontech, Palo Alto, CA) was used for the identification of
positively transfected cells.
Tissue Culture. CHO cells were plated at a density of 5.6 ? 104
cells per well on coverslips in 24-well plates 24 h before trans-
fection. Cells were treated for 4 h in serum-free medium (F12
nutrient medium with 10 mM Hepes) with a total of 1.2 ?l of
Lipofectamine (Invitrogen, Carlsbad, CA) and 0.28 ?g of DNA
per well (0.14 ?g of both EGFP and potassium channel cDNA).
Cells were briefly washed in MEM with Earle's salts containing
25 mM Hepes and 0.01% BSA. After transfection, cells were
maintained in F12 medium containing FBS at 37°C, 5% CO2for
48 h before recordings.
Cortical neurons were prepared from embryonic day 16 rat
embryos and grown in 6-well plates according to McLaughlin et
al. (8). Cultures were exposed to drug treatment procedures at
Fig. 5.
nonconducting Kv2.1(W365C, Y380T) and 24 h later exposed to 30 ?M DTDP
expressed as a percent of vehicle-treated control. Note that DTDP induced
?50% cell death in Kv2.1-expressing CHO cells but was not lethal to either
the mean ? SEM viability from three separate, independent experiments (*,
P ? 0.05; ANOVA/Dunnett).
S800A mutation blocks Kv2.1-mediated apoptosis. CHO cells were
Redman et al.
February 27, 2007 ?
vol. 104 ?
no. 9 ?
Page 5
25–29 days in vitro. Cells were briefly washed in MEM with
Earle's salts containing 25 mM Hepes and 0.01% BSA and
maintained in D2C growth medium until harvesting. Cells were
harvested 3 h posttreatment in lysis buffer [1% Triton X-100/
0.1% SDS/0.25% Na deoxycholate/50 mM Hepes/150 mM NaCl/
protease inhibitor mixture (Roche Diagnostics, Indianapolis,
IN), pH 7.5] after two washes with PBS. Cell lysate samples were
combined in a 1:1 ratio with reducing sample prep buffer and
incubated for 5 min at 100°C to denature proteins before gel
Drug Treatment. The apoptotic stimulus for the electrophysiolog-
ical experiments in CHO cells consisted of a 5-min treatment
with 25 ?M DTDP at 37°C, 5% CO2. The DTDP-containing
solution was then removed and replaced with fresh F12 medium
containing 10 ?M 1–3-boc-aspartyl (Ome)-fluoromethyl-ketone
(BAF), a broad-spectrum cysteine protease inhibitor. BAF was
necessary to maintain cells viable for electrophysiological re-
cordings because Kv2.1-expressing cells are highly susceptible to
DTDP-induced apoptosis (13). Cells were subsequently main-
tained in BAF-containing medium, and electrophysiological
recordings were performed ?3 h after oxidative injury. For
channel insertion experiments, cells were first treated with 4 ?M
MTSET for 10 min to covalently block all Kv2.1(I379C) or
Kv2.1(I379C, S800A) channels present on the plasma membrane
surface before the usual DTDP exposure (14). The apoptotic
stimulus for the biochemical experiments on rat primary cortical
neurons consisted of a 10-min treatment with 100 ?M DTDP at
37°C, 5% CO2 (7, 8). Cortical neurons were pretreated with
either vehicle or the specific p38 MAPK inhibitor SB-293063 (20
?M for 10 min).
Electrophysiological Measurements. Current recordings were per-
formed on EGFP-positive cells by using the whole-cell patch clamp
configuration technique as described (8). We observed that 97% of
GFP-positive CHO cells cotransfected with Kv2.1 had measurable
K-gluconate, 10 mM KCl, 1 mM MgCl2, 1 mM CaCl2? 2H2O, 10
mM Hepes; pH adjusted to 7.2 with concentrated KOH; 0.22 mM
sucrose. The extracellular solution contained 115 mM NaCl, 2.5
mM KCl, 2.0 mM MgCl2, 10 mM Hepes, 0.1 mM 1,2-bis(2-
aminophenoxy)ethane-N,N,N?,N?-tetraacetate acid, 10 mM D-
glucose, and 0.1 mM tetrodotoxin; pH was adjusted to 7.2. Mea-
surements were obtained under voltage clamp conditions with an
Axopatch 1-D amplifier (Axon Instruments, Foster City, CA) and
pClamp software (Axon Instruments) using 2- to 3-M? electrodes.
Recording electrodes were pulled from 1.5-mm borosilicate glass
pipette puller (Sutter Instruments, Novato, CA). Series resistance
at 2 kHz and digitized at 10 kHz with Digidata software (Axon
Instruments). Potassium currents were evoked with incremental
15-mV voltage steps to ?35 from a holding potential of ?70 mV.
To determine current density values, steady-state current ampli-
and normalized to cell capacitance. All data are expressed as
mean ? SEM, and statistical analysis was performed with InStat
software (GraphPad, San Diego, CA).
Generation of Phospho-Specific Antibody. To generate an antibody
specific for the phosphorylated p38 MAPK consensus se-
quence of Kv2.1 (pKv2.1; SI Fig. 6), we synthesized the peptide
C-KNHFESSPLPTS(p)PKFLR (Tufts University Core Facil-
ity, Boston, MA). The HPLC-purified peptide was conjugated
to keyhole limpet hemocyanin (Pierce, Rockford, IL) with
Sulfo-Link (44895; Pierce) following the manufacturer's pro-
tocol. Antiserum was generated in New Zealand White rabbits
at Covance (Princeton, NJ). After prebleed screening, animals
were initially injected with 1.5 mg of immunogen, followed by
three monthly boosts of 0.75 mg and three monthly boosts of
0.5 mg. The crude serum was affinity-purified by using an
Immunopure (A) IgG Purification Kit (Pierce) according to
the methods described by the manufacturer, and the resulting
product was used in the experiments reported here. A com-
mercially available Kv2.1 polyclonal antibody (Alomone
Labs), which was not targeted to the p38 site, was used as a
control in immunoblots.
Kv2.1(S800A)-expressed protein from transfected CHO cells
were immunoprecipitated by incubating cell lysates with an
anti-myc-tag rabbit polyclonal antibody (40 ?g/?l) (MBL
International Corp., Woburn, MA) followed by a protein A/G
PLUS-Agarose immunoprecipitation reagent (Santa Cruz
Biotechnology, Santa Cruz, CA). The immunopurified sub-
strate was incubated in 15 ?l of kinase buffer (25 mM Hepes,
pH 8.0/2 mM DTT/0.1 mM vanadate), 15 ?l of Mg/ATP (50
mM MgCl and 50 ?M ATP), and 50 ?g activated p38? (Roche
Protein Expression Group, Indianapolis, IN) for 1 h at 30°C.
Reactions containing kinase buffer alone and kinase buffer
plus Mg/ATP were used as controls. The reaction was stopped
with sample preparation buffer (625 mM Tris/25% glycerol/2%
SDS/0.01% bromophenol blue/5% ?-mercaptoethanol) and
incubated at 100°C for 5 min before SDS/PAGE and immu-
noblotting. Immunoblotting was performed with the pKv2.1
antibody (1:12,000) and a commercially available polyclonal
Kv2.1 antibody (1:3,000) (Alomone Labs).
KinaseAssay. Polymyc-Kv2.1- and polymyc-
Electrophoresis and Immunoblotting. SDS/PAGE was carried out
by standard procedures using the Mini Protean 3 System
(Bio-Rad, Hercules, CA). Equal amounts of cell lysate were
separated by reducing 6% or 10% SDS/PAGE gels. Separated
protein bands were transferred onto a 0.2-?m nitrocellulose
membrane (Bio-Rad). The membranes were then blocked with
1% BSA in PBS with 0.05% Tween 20 (PBST) at room
temperature for 1 h and probed with the appropriate primary
antibodies diluted in PBST. Blots were then incubated with
goat secondary antibody conjugated to HRP at room temper-
ature for 1 h. Blots were visualized with a SuperSignal
CL-HRP Substrate System (Pierce) and exposed to BioMax
films (Kodak, New Haven, CT). In the cortical culture exper-
iments, optical density measurements (Scion Image software,
National Institutes of Health) were taken of both pKv2.1
immunoreactive bands, normalized to their respective Kv2.1
immunoreactive bands, and the resulting values were pooled.
Viability Assays. CHO cells were plated and transfected with
EGFP and pRBG4 vector plus Kv2.1, Kv2.1(S800A) or
Kv2.1(W365C/Y380T) according to the protocol described
above, with 0.28 ?g of DNA added per well (0.14:0.13:0.0028
?g; EGFP/empty vector/potassium channel cDNA). Twenty-
four hours after transfection, cells were exposed to either 30
?M DTDP or vehicle for 15 min at 37°C, 5% CO2. Twenty-four
hours after treatment, counts of GFP-positive cells were
obtained by a person blinded to the experimental treatment
groups from 15 fields with a ?20 objective per coverslip; three
coverslips were counted per condition in three independent
We thank D. Jimenez for preliminary electrophysiological measure-
ments, M. Aras for helpful suggestions, and Drs. J. Trimmer, K.
Takimoto, J. Nerbonne, and S. Korn for the gifts of plasmids. This work
was supported by National Institutes of Health Grants NS043277 (to
E.A.) and HL080632 (to E.S.L.) and Mental Retardation Research
Center Grant HD018655 (to P.A.R. and L.H.).
3572 ?
www.pnas.org?cgi?doi?10.1073?pnas.0610159104Redman et al.
Page 6
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vol. 104 ?
no. 9 ?


text:
(Martes pennanti)
Webpage Author: Michael Sedon
The fisher one of the largest
members of the weasel family disappeared from
Fishers in the state of
Fishers are
similar to the much smaller weasels. They have a long body with short legs.
Adults range in length from about 2.5 to 4 feet. Males weigh about 7 to 13
pounds, females about 3 to 5.5 pounds. The fisher has a broad and flat
head with a sharp
pronounced muzzle. The eyes on a fisher face forward and the ears are
rounded (Pack and Cromer 1981). The fur on the fisher ranges from 1 inch
to 2.75 inches long and the color ranges from light brown to dark brown.
It also has a large bushy tail. Fishers have five toes on all four feet
with retractable claws like a cat. They can also rotate their hind paws almost
180 degrees allowing them to come down trees head first like a squirrel and
grasp limbs (Wallace 1985). Fishers are predators with a diverse diet that
includes birds, porcupines, snowshoe hare, squirrels, mice, shrews, voles,
reptiles, insects, deer carrion, vegetation and fruit. The breeding season for
fisher begins in February and ends in the middle of April. Litter sizes
can range anywhere from 1 to 4 but have been known to get as high as 5 or 6
(Powell 1993).
The primary threat is the
loss and fragmentation of fisher habitat, which is due to timber harvest,
roads, urban development, recreation, and stand-replacing fire (Berg
1982). Other factors cited include poaching and incidental capture and
injury, predation, mortality by vehicle collision, limited population size, and
isolation of populations (Halfpenny 1995).
Habitat Requirements:
The requirements for habitat for the fisher are
continuous forested areas, since they very rarely ever travel into unforested areas (Hagmeier 1956). It has long
been thought that coniferous forests were needed for the survival of fishers
but it seems that, in
Kingdom: Animalia
Phylum: Chordata
Class: Mammalia
Order: Carnivora
Family: Mustelidae
Genus: Martes
Species: pennanti
Common Names: fisher, tree
fox, and black cats
Berg, W. E. 1982. Reintroduction of fisher, pine marten and river otter.
Pages 159–173 in G. C. Sanderson,
Hagmeier, E. M. 1956. Distribution of marten and fisher in
Halfpenny, J. C., R. W.
Thompson, S. C. Morse, T. Holden, and P. Rezendes. 1995. Snow tracking. Pages
91–163 in
Zielinski and T. E. Kucera,
eds. American marten, fisher, lynx, and wolverine: survey methods for their


text:
Thursday, December 11, 2008
A Letter
6 comments:
@ rigtenzin--I absolutely agree about the license stickers and old bike shop stickers, I love those whenever I see them. But crusty old tape doesn't float my boat in the same way. Also, I probably wouldn't mind so much if the fenders were in decent shape paint-wise and I didn't have to chip that old tape off in order to re-paint them.
ReplyDelete@ R:B--Several California cities, including Davis and Los Angeles, currently require bike licenses, and the practice was much more widespread in the 1960s and 1970s, from what I understand. Nationwide, I believe a number of other cities also require licenses, but I don't believe there is any such requirement at the state level. My Columbia had the partial remnants of what I believe was a reflective registration sticker on the rear fender before I painted it, but it wasn't worth saving.
I've got mixed feelings about bicycle registration. On the one hand, it's a step toward recognizing the bicycle as transportation, and helps with recovering stolen bicycles, but on the other hand, it smacks of government bureaucracy sticking its money-grubbing hands in where it doesn't need to be.


text:
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Civil War letters shared
on some of those that served
If you would like to submit Ontario County data to this site please contact me.
Copyright resides with the contributor. Transcribed by Dianne Thomas.
Return to Home Page Return to CW Military Index page
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148 New York Infantry is doing garrison duty here in
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Esmans Vacations
Esmans is near a range of colorful Seine-et-Marne's more popular attractions. The Church of the Assumption of Esmans and Montereau-Fault-Yonne Park are just two of the popular places to visit. Two other popular spots that are located close by Esmans are the Church of Montereau-Fault-Yonne and the Golf Club Montereau La Forteresse.
To really understand Seine-et-Marne, visit one of its museums and theaters. Browse the interactive and traditional museums and collections in the Musee de Sens in Sens.
If you love to play golf, take a trip to the Fontainebleau Golf Club in Fontainebleau, the Fontenailles Golf Club in Fontenailles and the Bois-le-Roi Golf Club in Paris. It's a perfect way to get out and enjoy the outdoors.
When you're ready to rest up after checking out the charms of Esmans, check in to one of the area's many short and long stay accommodation options. The Novotel Fontainebleau Ury, the Hotel Première Classe Sens Nord Saint Clément, Saint-Clement and the Hotel Kyriad Nemours are just some of the most popular accommodation options here.


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Additional Physical Format: | Print version: Steele, Earl N. Rise and decline of the Olympia oyster. [Elma, Wash., Fulco Publications, 1957] (DLC) 58017211 (OCoLC)9470270 |
Material Type: | Document, Internet resource |
Document Type: | Internet Resource, Computer File |
All Authors / Contributors: |
E N Steele |
OCLC Number: | 271187741 |
Reproduction Notes: | Electronic reproduction. [S.l.] : Biodiversity Heritage Library, 2010. DcWaBHL |
Description: | 1 online resource (126 pages) illustrations |
Details: | Master and use copy. Digital master created according to Biodiversity Heritage Library Digital Imaging Specifications. |
Other Titles: | Olympia oyster |
Responsibility: | by E.N. Steele for the Olympia Oyster Growers Association. |
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It's a very fancy looking watch and can be used with a casual or elegant outfit! My husband really liked it!
This is the third Invicta I have purchased. This one is by fat=r the best-looking one. The blue face is so masculine and rich looking everyone will think your special man is James Bond
This is the second time I bought this watch in 8 years. so you can see that I like it a lot. the only thing is...I wish that it still had the automatic movement that came on the first watch, it had the glass back were you could see the movement of the watch and you never had to replace a battery. which was pretty cool!! but other then that it's still a very nice watch! even with the Swiss movement.
The watch looks good and keeps good time. The first day it lost 6 minutes but after that it didn't lose time. It is stainless steel and therefore rugged.. It is a good" wear everyday" watch.
I bought this watch for my husband's birthday. I've really enjoyed him getting used to the look of it, the weight of it (quite heavy) and pausing to admire it from time to time. I feel it was a very good value for the money and though it is new now, I anticipate it will wear well throughout the years.
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¹ "Measuring Up 2008: The National Report Card on Higher Education." The National Center for Public Policy and Higher Education, 2008. p. 8.
² Kroll, Andy. "Shut Out: How the Cost of Higher Education is Dividing Our Country." April 3, 2009 ; TomDispatch.com. Retrieved November 17, 2009: commondreams.org/view/2009/04/03-3.
Read the full story at http://www.prweb.com/releases/Eye_Care/Ophthalmology/prweb3349004.htm.
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Tuesday, October 25, 2011
Tuesday Talkback: Greatest Horror Sequels
For the purposes of this discussion, assume "so bad they're good" movies are perfectly acceptable.
6 comments:
I'm sure I'll receive a fair amount of flack for this, but how about Scream 2 and Saw 2?
Scream 2 is essentially just another story, it's killers are largely unimportant though when you look at the way the movie is structured around the survivors from the original. And there are some truly stressful sequences, such as Courtney Cox quietly running from Ghostface in a recording studio. Love it.
Saw 2 just has a terrific structure. The opening is a standard trap that simultaneously reacquaints you with Jigsaws methods while also being integral to the plot. The direction actually isn't as frantic as the later sequels would become, and the set of the run down house is terrific. Not the greatest movie in the world, but a damn good sequel. It also has an ending that you can pretend caps off the entire series. No one needs Saw III - VIIExorcist 2 for Showgirls level unintentional comedy.
Halloween H20 - terrible title, but has a fun character arc for Jamie Lee Curtis going from alcoholic to axe-wielding warrior.
Dawn of the Dead (sequel to Night of)
Friday the 13th Part 6 - Where a director finally realized how campy the whole series was and just had fun with it.
Psycho 3. Part 2 was overplotted, but not half bad. Part 3 just really lays on the weird and it better than anyone gives it credit for.Whole day marathon anyone?
1. Underworld: Evolution - Start with horror/action
2. Noriko's Dinner Table - Confusing/philosophical horror
3. 28 Weeks Later... - More violent action
4. The Silence Of The Lambs - Then bring it back down a notch
5. Dawn Of The Dead ('78)
6. Evil Dead II - Horror/Comedy to keep you awake
7. The Embodiment Of Evil - End with crazy cult/gore from Brazil


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Research/Internship Opportunity Detail Page
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NASA-ESMD internships
Primary Institution: | New York Space Grant Consortium |
States: | AL, CA, FL, MD, OH, TX |
Job Duration: | 10 weeks |
Class Requirement: | Completed Sophomore Year |
Minimum GPA: | 3.0 out of 4.0 |
Position Type: | Research |
Discipline(s): | Electrical Engineering Mechanical Engineering Other Engineering Interdisciplinary Physics: Biological Physics; Quantum Information; Etc. Various sciences, Various engineering |
Description: | Funding from the NASA Exploration Systems Mission Directorate (ESMD) allows the New York Space Grant Consortium to provide $5,000 stipends and $1,000 travel/relocation allowances for 10-week summer internships at various NASA centers, such as Ames Research Center, Glenn Research Center, Goddard Space Flight Center, Johnson Space Center, Kennedy Space Center, and Marshall Space Flight Center. Students will engage in projects that specifically address the ESMD goal of creating new capabilities and technologies that enable sustained and affordable human and robotic space exploration. Design, engineering, and research opportunities are available in the following areas: spacecraft, propulsion, lunar and planetary surface systems, and ground operations. The ESMD internship opportunities are listed on the following page: NASA-ESMD Space Grant Internships. Be sure to check the timeframe of the opportunities, as NY Space Grant support is for summer 2007 internships only. ELIGIBILITY: To receive NY Space Grant support, student must (1) be a United States citizen, and (2) be a New York state resident or attend a New York college/university. Additional eligibility requirements are listed at the top of the ESMD application form. APPLICATION: Apply directly to the NASA-ESMD Space Grant Internship Program via the following link: >>NASA-ESMD Space Grant Internship Application DEADLINES AND NOTIFICATION: February 16, 2007 is the direct application deadline. |
Stipend: | $5000 |
Benefits: | $1000 travel |
Application Deadline: | 02/16/2007 |
Job Website: | http://astro.cornell.edu/spacegrant/undergrad.html#esmd |
Research/Internship Opportunity Contact Information | |
Primary Contact: |
Erica Miles 517 Space Sciences Bldg. Cornell University Ithaca, NY 14853-6801 Email: firstname.lastname@example.com Phone: 607-255-2710 |