Strange behavior: New study exposes living cells to synthetic protein

Dec. 27, 2012 ? One approach to understanding components in living organisms is to attempt to create them artificially, using principles of chemistry, engineering and genetics. A suite of powerful techniques -- collectively referred to as synthetic biology -- have been used to produce self-replicating molecules, artificial pathways in living systems and organisms bearing synthetic genomes.

In a new twist, John Chaput, a researcher at Arizona State University's Biodesign Institute and colleagues at the Department of Pharmacology, Midwestern University, Glendale, AZ have fabricated an artificial protein in the laboratory and examined the surprising ways living cells respond to it.

"If you take a protein that was created in a test tube and put it inside a cell, does it still function," Chaput asks. "Does the cell recognize it? Does the cell just chew it up and spit it out?" This unexplored area represents a new domain for synthetic biology and may ultimately lead to the development of novel therapeutic agents.

The research results, reported in the advanced online edition of the journal ACS Chemical Biology, describe a peculiar set of adaptations exhibited by Escherichia coli bacterial cells exposed to a synthetic protein, dubbed DX. Inside the cell, DX proteins bind with molecules of ATP, the energy source required by all biological entities.

"ATP is the energy currency of life," Chaput says. The phosphodiester bonds of ATP contain the energy necessary to drive reactions in living systems, giving up their stored energy when these bonds are chemically cleaved. The depletion of available intracellular ATP by DX binding disrupts normal metabolic activity in the cells, preventing them from dividing, (though they continue to grow).

After exposure to DX, the normally spherical E. coli bacteria develop into elongated filaments. Within the filamentous bacteria, dense intracellular lipid structures act to partition the cell at regular intervals along its length. These unusual structures, which the authors call endoliposomes, are an unprecedented phenomenon in such cells.

"Somewhere along the line of this filamentation, other processes begin to happen that we haven't fully understood at the genetic level, but we can see the results phenotypically," Chaput says. "These dense lipid structures are forming at very regular regions along the filamented cell and it looks like it could be a defense mechanism, allowing the cell to compartmentalize itself." This peculiar adaptation has never been observed in bacterial cells and appears unique for a single-celled organism.

Producing a synthetic protein like DX, which can mimic the elaborate folding characteristics of naturally occurring proteins and bind with a key metabolite like ATP is no easy task. As Chaput explains, a clever strategy known as mRNA display was used to produce, fine-tune and amplify synthetic proteins capable of binding ATP with high affinity and specificity, much as a naturally occurring ATP-binding protein would.

First, large libraries of random sequence peptides are formed from the four nucleic acids making up DNA, with each strand measuring around 80 nucleotides in length. These sequences are then transcribed into RNA with the help of an enzyme -- RNA polymerase. If a natural ribosome is then introduced, it attaches to the strand and reads the random sequence RNA as though it was a naturally-occurring RNA, generating a synthetic protein as it migrates along the strand. In this way, synthetic proteins based on random RNA sequences can be generated.

Exposing the batch of synthetic proteins to the target molecule and extracting those that bind can then select for ATP-binding proteins. But as Chaput explains, there's a problem: "The big question is how do you recover that genetic information? You can't reverse transcribe a protein back into DNA. You can't PCR amplify a protein. So we have to do all these molecular biology tricks."

The main trick involves an earlier step in the process. A molecular linker is chemically attached to the RNA templates, such that each RNA strand forms a bond with its newly translated protein. The mRNA-protein hybrids are exposed to selection targets (like ATP) over consecutive rounds of increasing stringency. After each round of selection, those library members that remain bound to the target are reverse-transcribed into cDNA (using their conveniently attached RNA messages), and then PCR amplified.

In the current study, E. coli cells exposed to DX transitioned into a filamentous form, which can occur naturally when such cells are subject to conditions of stress. The cells display low metabolic activity and limited cell division, presumably owing to their ATP-starved condition.

The study also examined the ability of E. coli to recover following DX exposure. The cells were found to enter a quiescent state known as viable but non-culturable (VBNC), meaning that they survived ATP sequestration and returned to their non-filamentous state after 48 hours, but lost their reproductive capacity. Further, this condition was difficult to reverse and seems to involve a fundamental reprogramming of the cell.

In an additional response to DX, the filamentous cells form previously undocumented structures, which the authors refer to as endoliposomes. These dense lipid concentrations, spanning the full width of the filamented E. coli, segment the cells into distinct compartments, giving the cells a stringbean-like appearance under the microscope.

The authors speculate that this adaptation may be an effort to maintain homeostasis in regions of the filamentous cell, which have essentially been walled off from the intrusion of ATP-depleting DX. They liken endoliposomes to the series of water-tight compartments found in submarines which are used to isolate damaged sections of the ship and speculate that DX-exposed cells are partitioning their genetic information into regions where it can be safely quarantined. Such self-compartmentalization is known to occur in some eukaryotic cells, but has not been previously observed in prokaryotes like E. coli.

The research indicates that there is still a great deal to learn about bacterial behavior and the repertoire of responses available when such cells encounter novel situations, such as an unfamiliar, synthetic protein. The study also notes that many infectious agents rely on a dormant state, (similar to the VBNC condition observed in the DX-exposed E. coli), to elude detection by antibiotics. A better understanding of the mechanisms driving this behavior could provide a new approach to targeting such pathogens.

The relative safety of E. coli as a model organism for study may provide a fruitful tool for more in-depth investigation of VBNC states in pathogenic organisms. Further, given ATP's central importance for living organisms, its suppression may provide another avenue for combating disease. One example would be an engineered bacteriophage capable of delivering DX genes to pathogenic organisms.

Share this story on Facebook, Twitter, and Google:

Other social bookmarking and sharing tools:


Story Source:

The above story is reprinted from materials provided by Arizona State University. The original article was written by Richard Harth.

Note: Materials may be edited for content and length. For further information, please contact the source cited above.


Journal Reference:

  1. Shaleen B. Korch, Joshua M. Stomel, Megan A. Le?n, Matt A. Hamada, Christine R. Stevenson, Brent W. Simpson, Sunil K. Gujulla, John C. Chaput. ATP Sequestration by a Synthetic ATP-Binding Protein Leads to Novel Phenotypic Changes inEscherichia coli. ACS Chemical Biology, 2012; : 121203123002005 DOI: 10.1021/cb3004786

Note: If no author is given, the source is cited instead.

Disclaimer: Views expressed in this article do not necessarily reflect those of ScienceDaily or its staff.

Source: http://feeds.sciencedaily.com/~r/sciencedaily/top_news/top_environment/~3/HOgFMtBFq6s/121227143001.htm

hangout todd haley kareem abdul jabbar miramonte elementary school mark jenkins super bowl commercials 2012 mia

Hawaii lieutenant gov. picked to fill Senate seat

Hawaii Lt. Gov. Brian Schatz speaks the state Capitol in Honolulu on Wednesday, Dec. 26. 2012 after Gov. Neil Abercrombie, right, announced he was appointing Schatz to fill the seat vacated by the late U.S. Sen. Daniel Inouye. (AP Photo/Audrey McAvoy)

Hawaii Lt. Gov. Brian Schatz speaks the state Capitol in Honolulu on Wednesday, Dec. 26. 2012 after Gov. Neil Abercrombie, right, announced he was appointing Schatz to fill the seat vacated by the late U.S. Sen. Daniel Inouye. (AP Photo/Audrey McAvoy)

FILE- In this Nov. 3, 2010 file photo, a supporter congratulates then-Lt. governor-elect Brian Schatz, center, as his wife Linda looks on at the Neil Abercrombie-Brian Schatz Hawaii governor's post election party in Honolulu. Schatz was appointed by Gov. Neil Abercrombie Wednesday, Dec. 26, 2012 to succeed the late U.S. Sen. Daniel Inouye. (AP Photo/Eugene Tanner, File)

Hawaii Lt. Gov. Brian Schatz, left, shakes hands with Gov. Neil Abercrombie at the state Capitol in Honolulu on Wednesday, Dec. 26. 2012 after the governor announced he was appointing Schatz to fill the seat vacated by the late U.S. Sen. Daniel Inouye. (AP Photo/Audrey McAvoy)

HONOLULU (AP) ? Hawaii's lieutenant governor and former head of President Obama's 2008 election effort there has been named the state's newest U.S. senator, a choice that went against the dying wish of revered longtime Sen. Daniel Inouye.

Gov. Neil Abercrombie appointed Brian Schatz to the post Wednesday, instead of Inouye choice U.S. Rep. Colleen Hanabusa.

The 40-year-old Schatz, a former state representative and onetime chairman of the state Democratic party, said his top priorities would be addressing global climate change, preserving federal funds used in Hawaii for things like defense spending and transportation, and getting federal recognition for Native Hawaiians for forming their own government, similar to many Indian tribes.

Schatz, who ran with Abercrombie for the state's top two offices in 2010, beat out Hanabusa and Esther Kiaaina, a deputy director in the state Department of Land and Natural Resources.

The three candidates were selected by state Democrats Wednesday morning from a field of 14. The candidates briefly made their cases before the state party's central committee.

"No one can fill Sen. Daniel K. Inouye's shoes, but what we want to do today is find the right person to walk in his footsteps," Schatz told the committee.

"I want to be your senator because I believe Hawaii needs seniority, and we need someone who can build it up over decades and decades. And I pledge to you, if I'm given this opportunity and obligation to serve, I will try to make this my life's work and rebuild the tenure that Hawaii so desperately needs," he said.

The White House said Schatz would fly to Washington Wednesday night with Obama, who was returning from his Hawaii Christmas vacation early as Congress considers what to do about the so-called fiscal cliff.

Schatz could be sworn in as early as Thursday, which would make him Hawaii's senior senator. Hawaii's other senator, Daniel Akaka, is retiring at the end of this Congress after 22 years.

Even before winning the 2010 general election, Abercrombie expressed faith in Schatz, saying he would put him in charge of attracting more private and federal investment in Hawaii. Other responsibilities included leading the state's clean energy efforts and Asia-Pacific relations.

"To the people of Hawaii, I can assure you this: I will give every fiber of my being to doing a good job for the state of Hawaii," Schatz told a news conference in Honolulu. "We have a long and perhaps difficult road ahead of us, but we can succeed if we work together. I understand the magnitude of this obligation and this honor, and I won't let you down."

Inouye, by far Hawaii's most influential politician and one of the most respected lawmakers in Washington after serving five decades in the Senate, died last week of respiratory complications at the age of 88. He sent Abercrombie a hand-signed letter dated the day he died, saying he would like Hanabusa to succeed him, calling it his "last wish."

Four days after eulogizing Inouye in the courtyard of the Hawaii Capitol, Abercrombie said he had to consider more than just Inouye's wishes in filling his seat.

"Of course Sen. Inouye's views and his wishes were taken into account fully, but the charge of the central committee, and by extension then myself as governor, was to act in the best interests of the party ... the state and the nation," Abercrombie said.

"The law makes explicitly clear, as do the rules of the Democratic Party, that while everyone's voice is heard and everyone's view is taken into account, nonetheless, no one and nothing is preordained."

Under state law, the successor had to come from the same party as the prior incumbent. An Abercrombie spokeswoman said the governor did not feel any political pressure from within his party to make the choice he made.

"While we are very disappointed that it was not honored, it was the governor's decision to make," Jennifer Sabas, Inouye's chief of staff, said in a statement. "We wish Brian Schatz the best of luck."

Selecting Hanabusa, 61, would have required a special election in Hawaii's 1st Congressional District. Last time that happened, Hanabusa lost to Republican Charles Djou because of a winner-take-all format that split votes between Democrats.

Abercrombie said the possibility of a special election was a factor, as well as Hanabusa's "key position" on the House Armed Services Committee. The governor said she was on her way toward establishing a senior position on that panel, and it's important for Hawaii ? with its four-member delegation ? to establish seniority in both chambers.

Schatz will serve until an election is held in 2014. He said he will run for re-election to try to keep the Senate seat until 2016 ? the end of Inouye's original term ? and would run again for Senate in 2016 if given the chance.

Hanabusa congratulated Schatz in a statement.

"Having served as chair of the Hawaii Senate Judiciary Committee when the succession law was passed, I fully respect the process and the governor's right to appoint a successor," she said.

Last week, when Abercrombie announced he would not seek the seat himself, his spokeswoman Louise Kim McCoy said Abercrombie intends to make his current post as Hawaii governor the last office of his political career.

Inouye would be "very happy" with the choice, Hawaii Democratic Party chairman Dante Carpenter said. Schatz has less experience than some older politicians in the Senate but he will be building seniority, which is "critical" to the state of Hawaii, he said.

"In the words of Sen. Dan Inouye ? invoked more than once ? seniority in the United States Congress is everything," Carpenter said.

Senate Majority Leader Harry Reid had urged Abercrombie to name Inouye's successor before the end of the year. The next Congress begins Jan. 3.

Democratic Rep. Mazie Hirono, D-Hawaii, was elected in November to succeed Akaka.

First in line to replace Schatz as lieutenant governor is Senate President Shan Tsutsui, who said he planned to discuss the prospect with his family before deciding.

___

Becky Bohrer can be reached on Twitter at http://twitter.com/bbohrerap.

___

Associated Press writer Audrey McAvoy contributed to this report.

Associated Press

Source: http://hosted2.ap.org/APDEFAULT/386c25518f464186bf7a2ac026580ce7/Article_2012-12-27-US%20Senate-Hawaii/id-888ae319a06044509b89cd1e2539916e

Hurricane Sandy new jersey atlantic city ocean city maryland Nexus 7 KDKA Pumpkin Carving Ideas

No deal in sight as deadline for fiscal deal nears

President Barack Obama waves as he boards Air Force One to return to Washington, at Honolulu Joint Base Pearl Harbor-Hickam in Honolulu, after spending Christmas with his family in Hawaii, Wednesday, Dec. 26, 2012. (AP Photo/Gerald Herbert)

President Barack Obama waves as he boards Air Force One to return to Washington, at Honolulu Joint Base Pearl Harbor-Hickam in Honolulu, after spending Christmas with his family in Hawaii, Wednesday, Dec. 26, 2012. (AP Photo/Gerald Herbert)

President Barack Obama and first lady Michelle Obama arrive to visit with members of the military and their families in Anderson Hall at Marine Corp Base Hawaii, Tuesday, Dec. 25, 2012, in Kaneohe Bay, Hawaii. The first family is in Hawaii for a family holiday vacation. (AP Photo/Carolyn Kaster)

President Barack Obama waves from the window of his motorcade vehicle as he returns from golf and a walk on the beach with his family at Marine Corp Base Hawaii, in Kailua, Hawaii, Wednesday, Dec. 26, 2012. The president and the first family are in Hawaii for a family holiday vacation. (AP Photo/Gerald Herbert)

President Barack Obama waves to base peronnel before boarding Air Force One to return to Washington, at Honolulu Joint Base Pearl Harbor-Hickam in Honolulu, after spending Christmas with his family in Hawaii, Wednesday, Dec. 26, 2012. (AP Photo/Gerald Herbert)

President Barack Obama greets base visitors and personnel before boarding Air Force One to return to Washington, at Honolulu Joint Base Pearl Harbor-Hickam in Honolulu, after spending Christmas with his family in Hawaii, Wednesday, Dec. 26, 2012. (AP Photo/Gerald Herbert)

(AP) ? Lawmakers are engaged in a playground game of "who goes first," daring each political party to let the year end without resolving a Jan. 1 confluence of higher taxes and deep spending cuts that could rattle a recovering, but-still-fragile economy.

President Barack Obama returns from Hawaii Thursday to this increasingly familiar deadline showdown in the nation's capital, with even a stopgap solution now in doubt.

Adding to the mix of developments pushing toward a "fiscal cliff," Treasury Secretary Timothy Geithner informed Congress on Wednesday that the government was on track to hit its borrowing limit on Monday and that he would take "extraordinary measures as authorized by law" to postpone a government default.

Still, he added, uncertainty over the outcome of negotiations over taxes and spending made it difficult to determine how much time those measures would buy.

In recent days, Obama's aides have been consulting with Senate Democratic Leader Harry Reid's office, but Republicans have not been part of the discussions, suggesting much still needs to be done if a deal, even a small one, were to be struck and passed through Congress by Monday.

At stake are current tax rates that expire on Dec. 31 and revert to the higher rates in place during the administration of President Bill Clinton. All in all, that means $536 billion in tax increases that would touching nearly all Americans. Moreover, the military and other federal departments would have to cut $110 billion in spending.

But while economists have warned about the economic impact of tax hikes and spending cuts of that magnitude, both sides appear to be proceeding as if they have more than just four days left. Indeed, Congress could still act in January in time to retroactively counter the effect on most taxpayers and government agencies, but chances for a large deficit reduction package would likely be put off.

House Republican leaders on Wednesday said they remain ready to negotiate, but urged the Senate to consider or amend a House-passed bill that extends all existing tax rates. In a statement, the leaders said the House would consider whatever the Senate passed. "But the Senate first must act," they said.

Aides said any decision to bring House members back to Washington would be driven by what the Senate does.

Reid's office responded shortly after, insisting that the House act on Senate legislation passed in July that would raise tax rates only on incomes above $200,000 for individuals and $250,000 for couples.

Meanwhile, Obama has been pushing for a variant of that Senate bill that would include an extension of jobless aid and some surgical spending reductions to prevent the steeper and broader spending cuts from kicking in.

For the Senate to act, it would require a commitment from Senate Republican Leader Mitch McConnell not to demand a 60-vote margin to consider the legislation on the Senate floor. McConnell's office says it's too early to make such an assessment because Obama's plan is unclear on whether extended benefits for the unemployed would be paid for with cuts in other programs or on how it would deal with an expiring estate tax, among other issues.

What's more, House Speaker John Boehner would have to let the bill get to the House floor for a vote. Given the calendar, chances of accomplishing that by Dec. 31 were becoming a long shot.

Amid the standoff, Geithner advised Congress on Wednesday that the administration will begin taking action to prevent the government from hitting its borrowing limit. In a letter to congressional leaders, Geithner said accounting measures could save approximately $200 billion.

That could keep the government from reaching the debt limit for about two months. But if Congress and the White House don't agree on how to avoid the "fiscal cliff," he said, the amount of time before the government hits its borrowing limit is more uncertain.

"If left unresolved, the expiring tax provisions and automatic spending cuts, as well as the attendant delays in filing of tax returns, would have the effect of adding some additional time to the duration of the extraordinary measures," he wrote.

Whenever the debt ceiling hits, however, it is likely to set up yet another deadline for one more budget fight between the White House and congressional Republicans.

Initially, clearing the way for a higher debt ceiling was supposed to be part of a large deal aimed at reducing deficits by more than $2 trillion over 10 years with a mix of tax increases and spending cuts, including reductions in health programs like Medicare. But chances for that bargain fizzled last week when conservatives sank Boehner's legislation to only let tax increases affect taxpayers with earnings of $1 million or more.

Obama and his aides have said they would refuse to let Republicans leverage spending cuts in return for raising the debt ceiling. But Republicans say the threat of voting against an increase in the limit is one of the best ways to win deficit reduction measures.

Another potential showdown is pending. A renewed clash over spending could come in late March; spending authority for much of the government expires on March 27.

___

Follow Jim Kuhnhenn on Twitter: http://twitter.com/jkuhnhenn

Associated Press

Source: http://hosted2.ap.org/APDEFAULT/89ae8247abe8493fae24405546e9a1aa/Article_2012-12-27-Fiscal%20Cliff/id-c01bc8ee9ea449a7b0a5222396e26564

megamillions winning numbers lotto winner michael oher jerry lee lewis cesar chavez winning lotto numbers lottery tickets

TV Sex, Love, & Romance ? 2012?s Best & Worst Relationships ...

Looking for something to keep you cozy as 2012 comes to a chilly close? TVLine hereby kicks off its two-part review of the year in TV with this heartwarming (and perhaps other body parts-warming) rundown of the year?s most notable sex- and love-related moments on TV.

This gallery captures what and who we loved, hated and couldn?t shut up about for the past 12 months. Best Love Story? Most Kick-Ass Parent? Sex Scene That Made Us Most Uncomfortable? Yeah, it?s all in here.

Click through the gallery below, then hit the comments with your thoughts. Which of our picks do you like? Which would you swap? Coming Thursday: Part 2 of our review, including Most Disappointing Show, Most Underrated Actor, Best Show No One Was Watching, Wackiest Plot Twist and other categories.



Source: http://tvline.com/2012/12/26/tv-sex-scenes-love-romance-best-worst-television-relationships/

Django Unchained Led Zeppelin Ned Rocknroll Norman Schwarzkopf Avery Johnson netflix weather channel

Drink Recipe: Hibiscus Ginger Punch Recipes from The Kitchn | The ...

2012_12_20-hibiscus1.jpgMost punch recipes cater to a single crowd: drinkers or non-drinkers. This one is delicious either way. Whether you fizz it up with champagne or mineral water, hibiscus ginger punch will put a colorful sparkle in your celebrations.

2012_12_20-hibiscus2.jpg

When I started dreaming of a festive punch, I initially considered cranberry or pomegranate but, frankly, I was a bit tired of these ingredients, which appear in every other beverage this time of year. Instead, I turned to a holiday tradition from Jamaica, where a drink called "sorrel" is made from hibiscus blossoms. Throughout the world, dried hibiscus or roselle is steeped in water to make crimson-colored tisanes, juices, and punches, and in Jamaica especially the hibiscus is combined with ginger for a tart and pleasantly spicy beverage.

To make this punch, I mixed a rich hibiscus and ginger base with ginger ale and then either champagne, sparkling white wine, or sparkling mineral water. For the non-alcoholic version I recommend using a true mineral water (like Perrier) rather than other carbonated waters. Its subtle mineral flavor and more delicate bubbles are a sophisticated substitute for the alcohol. But use any fizzy water that works for you.

On the subject of substitutions, what if you can't find dried hibiscus blossoms (also known as "flor de jamaica" in Latin American markets)? Try making a strong brew with hibiscus tea bags; Celestial Seasonings and The Republic of Tea are two widely available brands.

2012_12_20-hibiscus4.jpg

Hibiscus Ginger Punch

Serves about 12

4 1/2 cups water
1 cup dried hibiscus blossoms
2-inch piece of ginger, thinly sliced
1 cup sugar
4 cups chilled ginger ale
2 750-mL bottles chilled champagne, sparkling white wine, or sparkling mineral water
Optional: ice, garnishes of mint, lemon or lime slices

Combine water, hibiscus blossoms, and ginger in a pot. Bring to a boil, then lower heat and simmer for 20 minutes. Turn off heat and stir in sugar. Let cool, strain, and chill. (If you like, save some of the plumped-up hibiscus blossoms for garnish.)

To make punch, stir hibiscus juice with ginger ale and champagne or mineral water. Garnish if desired and serve immediately.

Related: Recipe: Hibiscus-Earl Grey Iced Tea

(Images: Emily Ho)

Source: http://www.thekitchn.com/drink-recipe-hibiscus-ginger-punch-recipes-from-the-kitchn-181852

wizards of waverly place cedric benson playoff schedule charles addams pinewood derby cars republican debate tonight tinker tailor soldier spy

Paul Daley out of Bellator tournament after problems with visa arise

Paul Daley's next shot in MMA has hit a roadbump. The British fighter, who was cut from the UFC after he threw a cheap shot after the bell at Josh Koscheck in 2010, is out of Bellator's welterweight tournament. He can't enter the United States because of issues getting a work visa.

ESPN reports Daley was involved in a bar brawl and was charged with assault, which is the problem with the visa. Daley denies the brawl, and says via his Facebook page it was just a problem with his visa.

It seems as though (sp) a mass press release has gone out to the MMA media stating I was involved (in a) bar brawl, arrested, and cannot obtain a P-1 visa to compete in the USA. I am currently awaiting a decision on my visa, which is not expected until late April. Which means I am unable to compete in the season 8 Bellator tournament, as it starts in January. I will be fighting in Europe in the coming months, so I am able to stay active while awaiting a decision on my visa status. I was not involved in a bar brawl. This is 100-percent a false statement.

Previously, Daley has fought in the United States for Bellator, Strikeforce and the UFC without any visa issues.

Source: http://sports.yahoo.com/blogs/mma-cagewriter/paul-daley-bellator-tournament-problems-visa-arise-133424597--mma.html

secret service scandal shea weber greystone sidney crosby at the drive in alternative minimum tax modeselektor

How Do I Tell My Friend Her Place Smells Like Cat Pee? | xoJane

I recently visited the apartment of a friend for the first time and got a narsty whiff of excessive cat pee. I was only there for about 5 minutes to drop something off, and I don?t know her all that well, so I don?t even know if she has cats. It?s one thing to break it to a close friend that her apartment reeks. It?s another thing entirely to unload your opinions on an acquaintance/stranger whose emotional intelligence has yet to be revealed to you. I?m kind of a pussy when it comes to this sort of thing, so I?d rather not say anything at all and pray that I never have to go over there again. But is that really the best way to handle this situation?

My former cat, Joga (named after Bj?rk?s song, which matches her personality dead-on), now belongs to my parents in a feline-owner switcheroo stealthily pulled off by my mom. She once read in some crazy Russian newspaper that cats are very handy in lowering high blood pressure, and asked if she could ?borrow? Joga for a couple of weeks to test out her new ?scientific? discovery. I lovingly obliged, sending the cat off to Pennsylvania, and yada yada yada, she still lives there like a fat furry princess two years later.

image

Obligatory cat photo of Joga. (photo credit: Mom)

?

But when Joga still lived with me in my pathetically cramped apartment, I?m proud to say my living quarters were never cat-pee-scented. None of my friends ever told me either way or anything, so maybe I?m just as na?ve as the current cat pee offender. My lovely circle of friends can be brutally honest sometimes though (Recent comments I've received: ?Your hair is super fried, but you could sorta pull it off if you were dressed like a surfer chick,? ?I hate your outfit, nothing matches,? and ?I have never seen you NOT dress slutty.?), so I?m going to assume they would have told it to me straight.

Since we?re on the topic of pet odor, let?s say you have a cat and you want to get to the root of the problem. How does one keep the pet stank out of the house, anyway?

I consulted my friend, a crazy cat lady who has not one but TWO cats (a ballsy move for New York City, imho) for advice. In all the years I?ve known her, her apartments have always smelled so fresh and so clean you?d never even know she had cats (of course, you?d know if you were Facebook friends with her, bitch posts photos of her cats and other random cats like it?s her JOB). Here is what she told me:

1. Clean the litter box every day. If you?re lazy, do it AT LEAST every other day.
2. Use Arm and Hammer brand cat litter, it?s awesome.
3. Mix in an odor-neutralizing powder in the litter (Arm & Hammer makes this stuff, too).
4. Place an odor absorber near the litter thingy.

image

Mmmm, orange-scented urination.

?

?

5. Dump the entire box of litter and wipe down the litter box and any affected area as often as possible. Once a month MINIMUM.
6. You are what you eat. Feed your cats high quality food for nicer-smelling droppings.

image

Smelly cat, smelly cat, what are they feeding you?

?

?

7. Invest in litter box ?furniture.? It can be pricey, but so worth the feeling of someone coming in and asking, ?Where on earth is your litter box??

For the price, I?d expect a rhinestone-encrusted model to match my iPhone, but it?s much classier than looking at a cat toilet that doesn?t flush.

This is pretty basic advice if you ask me (except for #5, which I never did. It?s really gross and I hate it, but you should do it), but for some reason some people just don?t get it, or maybe their sense of smell isn?t that great and they think it?s all good or something.

Now, some cats are gonna be dicks and disrespect your authority by refusing to cover up their daily deeds in the litter box, or they?ll leave little presents lying around for you in your household. If you are one of these people, these rules don?t apply. All I can say is I?m sorry and don?t piss off your cat, whatever you do. Good luck.

If you?re paranoid about your place smelling off, sans or avec pets, just use common sense and bomb it with air fresheners. Just do me a favor and use fancy stuff (Yankee candles = NO) so it looks like you know how to accessorize rather than trying to hide something.

?

Anyway, back to my friend with the cat pee issue she is seemingly unaware of. I have thought long and hard about this and I still don?t have a solution. I could get all passive-aggressive-like on her and share this article on Facebook, hoping she sees it and magically realizes it?s about her. Do you have a better suggestion?

Source: http://www.xojane.com/relationships/how-do-i-tell-my-friend-her-place-smells-like-cat-pee

beltane capitals john edwards conocophillips octomom dan savage new world trade center

Clippers roll past Celtics 106-77

LOS ANGELES (AP) ? Matt Barnes tied his season high with 21 points off the bench, Chris Paul had 11 assists and the Los Angeles Clippers defeated Boston 106-77 on Thursday night, becoming the first team to win 15 consecutive games since the Celtics four years ago.

Fellow reserve Jamal Crawford added 17 points, Blake Griffin had 15 points and Caron Butler 14 to help the Clippers improve the NBA's best record to 23-6.

Kevin Garnett scored 16 points for Boston, which never led in dropping to .500 at 14-14 with its fifth loss in seven games. Paul Pierce and Jeff Green added 12 points each to go with 10 apiece from Jason Terry and Rajon Rondo.

Boston's winning streak extended to 19 games in 2008-09.

Barnes had the Clippers' first five points in a 9-0 run that opened the fourth quarter and boosted their lead to 90-67. Willie Green briefly played; otherwise the Clippers' other starters rested to close out the game.

The Celtics were held to 10 points in the final 12 minutes.

Garnett had six points in the third when the Celtics were narrowly outscored 22-20, but still trailed 82-67. The Clippers' 9-0 run pushed their lead to 21 points before Boston went on a 10-4 run, including 3-pointers by Courtney Lee and Rondo to end the quarter.

The game got chippy at times. In the third, Jared Sullinger was called for a flagrant-1 foul for grabbing the front of Griffin on his way up to dunk. Then Butler and Pierce emerged from a big mass both holding onto the ball, leading to a jump ball. In the first, Rondo was lying on the ball and Paul tried to take it out from under him on the sideline, leading to a testy moment.

The Celtics cut their deficit to four points early in the second quarter ? the closest they came after the opening quarter ? with consecutive 3-pointers by Terry and Lee before the Clippers' second unit rebuilt the double-digit lead the starters had created. Crawford scored 13 points for Los Angeles, highlighted by a fast-break floater and a rainbow 3. He scored the final four points to keep the Clippers ahead 60-47 heading into halftime.

The Clippers came roaring out of the opening tip, outscoring the Celtics 24-9.

NOTES: The Clippers are assured of finishing December as their best month ever with two games remaining, including the second half of a back-to-back at Utah. They've already surpassed the franchise record for most wins in a month, bettering the old mark of 12 shared by the 1974-75 Buffalo Braves and the 1991-92 Clippers (which included Celtics coach Doc Rivers). ... Boston fell to 5-9 on the road. ... The Clippers' last loss was on Nov. 26 at home against New Orleans. ... Justin Bieber (who had Paul's young son sitting on his lap), Billy Crystal and Olympic 200 sprint champion Allyson Felix were among the Clippers' 67th consecutive sellout crowd.

Source: http://news.yahoo.com/clippers-roll-past-celtics-106-77-062716724--spt.html

how to carve a turkey ipad 2 Belk Macys Thanksgiving Day Parade 2012 Turkey Cooking Times wal mart happy thanksgiving

Paul Daley out of Bellator tournament after problems with visa arise

Paul Daley's next shot in MMA has hit a roadbump. The British fighter, who was cut from the UFC after he threw a cheap shot after the bell at Josh Koscheck in 2010, is out of Bellator's welterweight tournament. He can't enter the United States because of issues getting a work visa.

ESPN reports Daley was involved in a bar brawl and was charged with assault, which is the problem with the visa. Daley denies the brawl, and says via his Facebook page it was just a problem with his visa.

It seems as though (sp) a mass press release has gone out to the MMA media stating I was involved (in a) bar brawl, arrested, and cannot obtain a P-1 visa to compete in the USA. I am currently awaiting a decision on my visa, which is not expected until late April. Which means I am unable to compete in the season 8 Bellator tournament, as it starts in January. I will be fighting in Europe in the coming months, so I am able to stay active while awaiting a decision on my visa status. I was not involved in a bar brawl. This is 100-percent a false statement.

Previously, Daley has fought in the United States for Bellator, Strikeforce and the UFC without any visa issues.

Source: http://sports.yahoo.com/blogs/mma-cagewriter/paul-daley-bellator-tournament-problems-visa-arise-133424597--mma.html

windows 8 preview leap year moratorium dwts season 14 cast leap day michigan primary results olympia snowe

blondelle seamanship: Simple Abundance Exercises Can Change ...

Article title: Simple Abundance Exercises Can Change Your Mindset
Article Category: Self-Improvement

5 free summer clipart illustration of a happy smiling sun Simple Abundance Exercises Can Change Your Mindset

When you feel more prosperous, you will lead life of improved health, happiness and prosperity. However, when most of us think about our finances, most of the time we don?t have thoughts of prosperity.
Continue reading this article?

Source: http://www.medicalguide.pro/3210/simple-abundance-exercises-can-change-your-mindset-4.html

Marissa Mayer Jon Lord weather.com Colorado shootings dark knight rises Aurora shooting James Eagan Holmes

Source: http://kaokely.posterous.com/simple-abundance-exercises-can-change-your-mi

sleigh bells meek sturgis sturgis whitney houston laid to rest daytona bike week mary kay ash

Source: http://binudsingh.blogspot.com/2012/12/simple-abundance-exercises-can-change.html

football score ron paul nevada buffalo chicken dip soul train nevada caucus ufc 143 what time does the super bowl start

Source: http://blondelle-seamanship.blogspot.com/2012/12/simple-abundance-exercises-can-change.html

PacSun apple store bestbuy bestbuy gamestop black friday deals Sephora