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Perry responded that he is taking the threat so seriously, he is creating a whole new office within the Energy Department to handle it.
The new Office of Cybersecurity, Energy Security and Emergency Response (CESER) will coordinate efforts to protect the electric grid -- and should the worst happen, respond to compromises.
Russian efforts to disrupt the 2016 presidential election through cyberattacks and fake news have been the subject of numerous news stories for more than a year. Less publicized, but also serious, are a series of remote intrusions Russians have made into U.S. power plants, which have been confirmed by the Department of Homeland Security and the FBI.
"Our energy infrastructure is under attack," said Sen. Maria Cantwell of Washington, the top Democrat on the Senate Energy and Natural Resources Committee who over the past several months has been loudly beating the drum on the Russian threat to the grid. "It's under cyberattack and we need to do much more to protect it as a national critical asset. Russia has proven its ability to disrupt the grid."
Sen. Lisa Murkowski (R-Alaska), chair of the committee, backed her Democratic colleague: "Know I share Senator Cantwell's concern on this. I want to make sure that DOE is cooperating with DHS and the FBI with implementation of actions in response."
After first alerting utility companies last June, the Department of Homeland Security and the FBI last week reported that Russian agents had gained the access needed to shut down or otherwise sabotage power plants. It was the first time the Trump administration, which issued new sanctions against Russians in the wake of the revelations, pointed the finger squarely at Russia for the energy infrastructure hacks.
Perry tried to reassure the senators the threat was on his radar. "There is a clear role DOE plays on cyber," Perry said. "We are committed to being as technically advanced as possible."
Even before the blockbuster report, the Energy Department has begun to lay the groundwork for the creation of a new office meant to address the heightened natural and man-made threats to the U.S. electric grid.
The department first announced it was creating CESER last month. To ensure the office works with scientists at the Energy Department's national laboratories dotted across the country, CESER will report directly to Mark Menezes, who as undersecretary of energy is the No. 3 official in the department. Perry said Tuesday that reporting relationship creates a "direct pipeline of information" back to him.
The Energy Department, already designated by Congress as the lead federal agency to work with utilities on making sure their systems protect against intrusion, can establish the new office and assistant secretary post to run it without additional congressional approval.
Now, the Trump administration is asking for $470 million to prevent and address cyberattacks on the energy sector, $93 million of which would be injected into the new office.
On Tuesday, Cantwell countered that is not enough. "I've called for a doubling," she said, "but I can see where I am wildly underfunding what is one of the most serious threats to us as a nation right now."
At least three times over the past year, Cantwell has called on the Trump administration to produce a risk assessment on Russia's capability to hack the grid.. "We are deeply concerned that your administration has not backed up a verbal commitment prioritizing cybersecurity of energy networks and fighting cyber aggression with any meaningful action," Cantwell and other Democratic senators wrote to President Trump last June.
At least three times over the past year, Cantwell has called on the Trump administration to produce a risk assessment on Russia's capability to hack the grid. So far there has been silence from the Trump administration on the request, her office said.
"Do you believe that we need a risk assessment as a nation?" Cantwell finally asked Perry at the hearing.
"I think that's going on as we speak," Perry responded. "We have three different areas in DOE that are focused on cyber and have been meeting and having these conversations."
Yet just last week, Perry expressed doubts the federal government as a whole is doing enough to check the threat.
"I'm not confident that the federal government has a broad strategy in place that is not duplicating," Perry told the House Appropriations subcommittee on energy and water issues, "or is least duplicative as it can be."
Concerns over Russian cyberattacks have animated congressional hearings recently. Facing intense grilling from Democrats on the Senate Armed Services Committee last month, Adm. Michael S. Rogers, who heads both the National Security Agency and U.S. Cyber Command, bluntly admitted "we're probably not doing enough" to counter Russia online.
"They haven't paid a price at least that's sufficient to get them to change their behavior," Rogers said. He added the president has not yet ordered Cyber Command to try to disrupt Russian threats.
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— One more time: Perry again repeated his reassurance that he's "not going anywhere" after reports last week that Trump was considering the energy chief to replace embattled Veterans Affairs Secretary David Shulkin. Perry told the Senate energy panel Tuesday that he plans to stay in his role.
— Trump's Arctic refuge story: During a National Republican Congressional Committee fundraiser yesterday, Trump explained how he lifted a ban on oil and gas drilling in the Arctic National Wildlife Refuge and told the story of how he realized the issue was "a big deal."
Trump said a friend in the oil business called him following the passage of the Republican tax bill, which included a provision for Arctic refuge drilling. "Is it true that you have ANWR in the bill?'" Trump said his friend asked. "I said: 'I don't know. Who cares? What is that? What does that mean?' ... What's the big deal if they did put it in?'"
Trump recalled his friend telling him "every single president" including former president Ronald Reagan had attempted to open up drilling there. "I said, 'You've got to be kidding, I love it now.' And after that, we fought like hell to get ANWR."
The truth: After Trump told the same tale at a congressional Republican retreat in February, Sen. Dan Sullivan (R-Alaska) countered by recalling exactly how he and Sen. Lisa Murkowski (R) made their pitch early in Trump's tenure. "We had the opportunity to brief the president last year," Sullivan told The Post's Paul Kane and Erica Werner last month. "It was early, like February or March. Over an hour, in the Oval, you know, that's a lot of time." He added that Trump was familiar with Alaskan issues because his grandfather worked on Chilkoot Trail, an 1890s route that led to the Yukon gold fields.
— BLM vs. NPS: The Bureau of Land Management did not comply with a request from the National Park Service to hold off on a lease sale planned for Tuesday. An Oct. 23 letter outlined concerns that oil and gas drilling activities near two national monuments (Hovenweep and Natural Bridges) and two national parks (Arches and Canyonlands) could adversely affect views and air, groundwater and sound quality. All 13 parcels were sold online as part of a broader sale, with the lease prices ranging from $3 to roughly $91 an acre, The Post's Juliet Eilperin reported.
— Pruitt's pricey travel: Two groups investigating Scott Pruitt's travel spending both released more details about how much the Environmental Protection Agency chief spent in past months on hotels and airfare. The travel costs for Pruitt's round-the-clock security detail exceeded $30,000 during his trip to Italy last year, according to documents the EPA released to the nonprofit watchdog Environmental Integrity Project. And the EPA turned over documents to House Oversight and Government Reform Committee Chairman Trey Gowdy (R-S.C.) late Tuesday detailing nearly $68,000 in newly disclosed travel costs, including for stays at high-priced hotels in New York City and Paris.
— A call for a workforce analysis: Other Republican lawmakers called on Pruitt to schedule a briefing with congressional staff on the EPA's plans to make cuts and reorganize the agency. Reps. Greg Walden (R-Ore.), John Shimkus (R-Ill.) and Gregg Harper (R-Miss.) sent a letter to Pruitt urging him "to assist us in understanding more about EPA's plans to reorganize the agency and how workforce analysis will factor into those plans." The letter also noted that the agency had not conducted a workforce analysis in more than two decades.
— Snow day: On the first official day of spring, the Mid-Atlantic and Northeast had yet another major nor'easter in the forecast, The Post's Angela Fritz reports. And in Washington, with the "biggest snowstorm of winter" expected to begin Wednesday morning, federal offices in D.C. are closed.
— McDonald's is trying to boost its brand by promising to go green: The restaurant chain — famous for slinging beef, one of the most carbon-intensive meats out there — is adding LED lights and efficient kitchen equipment to its restaurants and offices with the aim of reduce greenhouse gas emissions by 36 percent compared with 2015 levels by the year 2030, Bloomberg News reports.
— Tariffs' toll: Trump's tariffs on foreign aluminum and steel will add thousands of jobs at domestic producers, helping to offset labor losses in other industries, according to a new study from the Coalition for a Prosperous America. Yet the nonprofit group, which shares Trump's skepticism toward free trade, admitted the tariffs would slow the growth of the U.S. economy has a whole, shrinking it by "8/1000th of 1 percent of U.S. gross domestic product," according to Bloomberg News.
- The Senate Environment and Public Works holds a hearing on oversight of the Nuclear Regulatory Commission.
- The American Coalition for Ethanol holds the 10th annual DC Fly-In & Government Affairs Summit
- The House Natural Resources Subcommittee on Water, Power and Oceans holds a hearing.
- The House Natural Resources Committee holds a markup on various bills.
- The House Appropriations Subcommittee on Commerce, Justice, Science and Related Agencies holds a hearing on the 2019 budget for the National Oceanic and Atmospheric Administration.
- The House Oversight Subcommittee on National Security holds a hearing on bureaucratic challenges to hurricane recovery in Puerto Rico.
Coming Up
- The Senate Energy and Natural Resources Committee holds a hearing on the 2018 Western Water Supply Outlook and Bills Related to Water Infrastructure and Drought Resiliency on Thursday.
- The American Enterprise Institute holds an event on the future of infrastructure policy under Trump on Thursday.
- USTR Robert Lighthizer will testify before the Senate Finance Committee on the president's 2018 trade policy agenda on Thursday.
- The Wilson Center holds an event on "Linking China's Domestic and Global Energy Ambitions" on Thursday.
- Berkeley's Energy Institute at Haas holds its annual POWER Conference on Energy Research and Policy on Friday.
— The brink of extinction: Sudan, the world's last male northern white rhino, was euthanized Monday by a veterinary team in Kenya. Now just two female northern white rhinos remain, The Post's Max Bearak reports — Sudan's daughter and granddaughter. The rhino's death puts the species on the brink of extinction.


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Act II of Thomas's Faison's "Phase Change" at the Frederick Arts Council's Pop-Up Gallery (11 West Patrick Street) – (Hours are 10 AM-4PM Weekdays)
The last three photos are of Ashley Hoffman's photography series created in conjunction with Faison's work.
Info about the artists from Facebook: Artist Thomas Faison is a filmmaker and installation artist living and working in Frederick. His films have been featured in film showcases and festivals in Baltimore, Ann Arbor, Chicago, and Frederick. His installation artwork has been featured in artist run spaces throughout Baltimore and he is the co-founder of the Baltimore Student Film Showcase, now operating within the Maryland Film Festival.
Ashley Renee Hoffman is a film photographer born and raised in Frederick, Maryland. Her photographs serve as means to understand the human condition, spirituality and our relationship with Nature. Most of her work is made in the Appalachian region, exploring both the vast beauty and conflicts on an ethical and environmental level. Hoffman joined Faison as he produced the video opera, finding and photographing alternative perspectives of the event. Her work will be on display and for sale in the lobby and halls of the gallery building.
The exhibit is made possible by a grant from the National Endowment for the Arts (NEA) gallery and it is also sponsored by Market and Research Resources (M&RR). The space is provided in-kind by the Ausherman Family Foundation.

subset: commoncrawl


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On Tuesday, Dec. 5, Sports Illustrated presented Colin Kaepernick with the Muhammad Ali Legacy award. An honor the magazine says is intended to "celebrate individuals whose dedication to the ideals of sportsmanship has spanned decades and whose career in athletics has directly or indirectly impacted the world."
There to personally hand the award to Kaepernick was none other than Beyoncé, who said she was both "proud and humbled" to present the award and thanked the quarterback for his personal sacrifice.
"Colin took action with no fear of consequence or repercussion, only hope to change the world for the better," said the singer "To change perception, to change the way we treat each other, especially people of color. We're still waiting for the world to catch up."
The crowd seemed thrilled to see the two icons on stage together, but not everyone was happy. Well known right-wing political commentator Tomi Lahren, who has a history of creating incendiary tweets, decided it was the perfect opportunity to attack both Kaepernick and Beyoncé.
Police-hating Beyoncé presents police and America-hating Kappy with a "legacy" award. This is how far we've fallen. Wow. pic.twitter.com/KIe6IqEQey
— Tomi Lahren (@TomiLahren) December 6, 2017
And obviously, if you attack Queen Bey, you're gonna get stung.
The Beyoncé jealousy is real. 😂 Perhaps because she is a beautiful, successful, talented woman and isn't on her parent's insurance?
— Marcella~I'm speaking (@marcellalaluna) December 6, 2017
These two have more talent and integrity than you could ever dream of #jealous
— Craig R. Patrick (@craigtastic85) December 6, 2017
Racism is so American that when people protest racism, certain racists, I mean people assume those who protest hate America. Case and point. https://t.co/M0OSZiBNSD
— Francis Maxwell (@francismmaxwell) December 6, 2017
Girl, you need to be working on your resume instead of watching what Bey's doing … she's employed… go apply somewhere @TomiLahren
— Alex (@college_girrl) December 6, 2017
There are some pretty sweet burns in there, but perhaps it's best to let Lonnie Ali, Muhammad's widow, have the last word.
I am proud to be able to present this to Colin for his passionate defense of social justice and civil rights for all people. Like Muhammad, Colin is a man who stands on his convictions with confidence and courage, undaunted by the personal sacrifices he has had to make to have his message heard. And he has used his celebrity and philanthropy to benefit some of our most vulnerable community members.
H/T Someecards


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Written In The Stars Nappy Clutches
This personalised Nappy Clutch is the perfect accessory to help keep Mummas and Little Ones stay organised.
Part of the new limited edition collection of modern Mumma essentials, this play mat was designed in partnership with fellow Australian children's accessories brand Bella Buttercup and is part of the Written in the Stars collection.
4 products


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There are many benefits of forming limited
liability partnership with partners. One of the advantages is limited liability
preserves the personal assets of partners from the business debts. This is because
limited liability partnership is a separate legal entity to the members and
thus partners' assets will not be affected. Sherrie (2017) stated that it is
also more flexible in setting up the process by which the business is to be run
unlike company requires to follow fixed and complicated process. There is a
better flexibility in the business management could be allowed. Partners also have
the power and right to decide on how they contribute to business operations
individually. Moreover, it has less formality and regulations to follow about the
management and structure of business compare to a company. Limited liability
partnership also can enjoy perpetual succession. That is, if one of the
partners passed away, retired or leave the partnership, the partnership still
can be continued to exist and operate. Aaron (2015) said that it is also more
simple and cheaper to incorporate and maintain because partners are not
required to have Annual General Meeting (AGM), hire a company secretary and
present audited financial reports. In addition, it is managed and controlled by
compliance officers or partners who responsible for decision making and to
vote. Lee (2015) mentioned that small businesses are likely to use limited
liability partnership.
limited liability partnership also has disadvantages. That is, it is required
to disclose accounts to the public. In this situation, they will have to
disclose the financial information and accounts to public. Furthermore, the
capital of limited liability partnership can only obtain from the members.
Thus, their relatives and close friends have to become members first if they
wish to raise capital from them.
Besides that, private company is also another business
structure which is suitable for the business of Sandy and Damian. Under Section
2(a) of Companies Act 2016, private company refers to any company which
immediately prior to commencement of the 2016 Act was a private company under
any previous repealed written law. Under Section 2(b), any company incorporated
as private company under the act, or Section 2(c), any company converted into
private company under Section 41.


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Love Hina (Episode 5) – A Trip to Kyoto
The time has come for the Tokyo U entrance exams Both have a past promise to keep, and have been studying hard. Will their hard work pay off?
"Wow, a Trip to Kyoto! Exciting!"
The fateful day has arrived – Tokyo U entrance exams.
Long story short – they both failed. After going together to see how they did, both Keitaro and Naru couldn't find their names on the board. They go to a restaurant for some drinks. Keitaro begins gushing about his promise, telling Naru that they can live happily together without going to Tokyo U, which prompts Naru to ask what he is on about.
Keitaro then finds out that Naru is not the girl he made the promise with, because 15 years prior Naru was only 2 years old. The girl he made the promise with would have to be closer to his own age, which is around 23 if I remember correctly. They part ways after Keitaro finds this out.
The next day, both Naru and Keitaro individually decide to go on a soul-searching trip to a traditional inn in Kyoto. They happen to get seats next to each other on the train. Keitaro bumps into Naru causing their glasses to fall onto the ground, and then while searching for them he accidentally sits on them, breaking them.
As a result, they both mistake each other for someone else, somehow. After spending time together in Kyoto, they both end up at the same inn, and are asked if they can share a room due to overbooking. Their repaired glasses are then delivered to them. They both end up getting over their post-failure sorrows, and the episode ends.
Episode Thoughts
Believe it or not, we didn't get any ecchi tropes this episode! The closest we got was Keitaro bumping into Naru on the train, but there was no accidental groping or anything. I guess Keitaro decided to be extra careful to not anger Naru after she failed her Tokyo U exam.
While it seems like a coincidence that they ended up going on the same trip, we find out that there was actually a flier for that specific inn in Kyoto lying around at the Hinata Apartments, so they both must have saw it and got the same idea.
There were some seemingly impossible moments this episode. How they didn't recognize each other's voice for an entire day doesn't make too much sense, for starters. And then when sleeping next to each other at the inn, Keitaro somehow managed to tie himself up in order to give Naru peace of mind, which seems impossible considering how he did it.
In Summary…
The biggest question we leave this episode with now is how the other girls at the Hinata Apartments handle these developments – first, they know that Keitaro and Naru went to Kyoto together, and second, they know Keitaro failed. Will they let him stay on as the Hinata Apartments manager?
The answer is clearly yes, because the anime would end otherwise. But I still wonder how things will play out in that regard.
Other Posts in the Series
- Season One
- Episode 1 – Welcome to the Hina Apartments
- Episode 2 – A New Guest
- Episode 3 – Motoko Returns
- Episode 4 – Keitaro's Academic Struggle
- Episode 5 – A Trip to Kyoto
- Episode 6 – Lost and Found
- Episode 7 – Introducing… Kentaro
- Episode 8 – Classic Gaming
- Episode 9 – Rent Left Unchecked
- Episode 10 – Su Confusion
- Episode 11 – Starstruck
- Episode 12 – Turtle Mayhem
- Episode 13 – Shinobu's Curiosity
- Episode 14 – Rat Race
- Episode 15 – Rekindled Love?
- Episode 16 – Teahouse Hinata
- Episode 17 – Ghostly Encounters on Vacation: Haunted Necklace?
- Episode 18 – Jealousy
- Episode 19 – The Crown Prince
- Episode 20 – Doll From A Bygone Era
- Episode 21 – Romantic Rivalry
- Episode 22 – Sisterly Conduct
- Episode 23 – Drama Ensues
- Episode 24 – Resolutions


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It did not take long for President Donald Trump to react to left-wing activist and vocal anti-Trump film director Spike Lee for his digs at the president during Oscar broadcasting.
In his acceptance speech for winning Best Director for "BlacKkKlansman," Lee said, "Let's all mobilize," he continued. "Let's all be on the right side of history. Make the moral choice between love versus hate. Let's do the right thing! You know I had to get that in there."
The Hollywood leftist also praised his ancestors, "who built our country."
His comments, a direct jab at the president, prompted this scathing response from Trump, who Tweeted Monday morning, "Be nice if Spike Lee could read his notes, or better yet not have to use notes at all, when doing his racist hit on your President, who has done more for African Americans (Criminal Justice Reform, Lowest Unemployment numbers in History, Tax Cuts,etc.) than almost any other Pres!"
Lee Not The Only Oscar Winner to Toss Shade on Trump
Though nominated before, this was the first Oscar for Lee. BlacKkKlansman is based on the true story of a black Colorado detective who infiltrated the Ku Klux Klan in Colorado in the 1970s.
Lee was far from the only Academy Award winner who referred to notes when delivering their acceptance speeches Sunday, nor was he the only attendee who took not so thinly veiled shots at the president during their time on stage.
In the opening monologue of the show, actress Maya Rudolph took a dig at Trump, lumping his reversal on having Mexico pay for a border wall in with the reversal of the Academy in creating a popular movie category and its decision not to have a host for this year's awards.
"Just a quick update for everybody in case you're confused," she said. "There is no host tonight. There won't be a popular movie category. And Mexico is not paying for the wall."
Trump, who was up particularly early to give his view on the Oscars, hasn't yet commented on Rudolph's jabs, nor the other so-called stars that made subtle digs at the Commander-in-Chief including Barbra Streisand and Javier Bardem.


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Reorganization Planning Team: Getting Started…
One of the most important components of the planning and creation of our newly reorganized Elementary Schools is the involvement of staff, parents, and community members in two different teams: The Reorganization Planning Team (RPT) and the Collaborative Design Team (CDT). The application process concluded last week and the teams are being launched this week.
Deb Landry, Trish Hill and Marc Smith will be having a joint meeting of the K-2 and 3-6 RPT groups on Tuesday October 14th from 6-8 pm in the Forestdale School Library. This meeting has two goals. First, is to plan for the first site visit, which has been organized for Friday, October 17th in Duxbury and East Bridgewater. This will be an all day excursion and it is understood that not all will be able to attend. A site visit will allow for various team members to observe and ask questions in a district that is also organized as two separate schools: Primary and Intermediate.
The second goal for the meeting is to create a meeting schedule for the entire team and participants are asked to bring a calendar.
The Administrative Team is very grateful to all those who volunteered for this important work. When making determinations for the group members, their goal was to create well balanced groups with balanced representation from each of the current school communities. A list of the Reorganization Planning Team is available here: RPT Contacts district


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PopMech posted an interesting story testing multiple padlocks against multiple stressors including a "Salt Fog" test for rust resistance and, it seems, a gun test.
Not to spoil things, but the Master lock was the only one that survived a gunshot to the gut. Everyone else shattered and ran away crying.


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Reviews (1)
I love how they mix well with my other plants
Helpful Gardening Tips
Goes Well With
Ostrich FernMSRP:Was:Now: $5.99
Fiddlehead FernMSRP:Was:Now: $8.99
Fern Plants Grab bag - 40 FernsMSRP:Was:Now: $74.99
Perennial Garden PackageMSRP:Was:Now: $49.99
Christmas FernMSRP:Was:Now: $8.99
Lady FernMSRP:Was:Now: $8.99
Fundraiser Fern Garden CollectionMSRP:Was:Now: $89.99
EvergreensMSRP:Was: $15.99Now: $13.99 - $499.99
Virginia CreeperMSRP:Was:Now: $8.99
Native GrassesMSRP:Was:Now: $8.99
Shipping Information
We ship all plants usps priority mail. They arrive to most locations within 2-3 days. We package all plants to retain moisture to up to 10 days in transit. All plants ships from our warehouses in Tennessee. All plants are grown and shipped from out Altamont (zip) 37301 location. We do drop ship for re-sellers also for those wanting to resell our plants.
How We Protect Your Plants For Transit
All plants are dug and immediately taken to our warehouse and tera-sorb moisture retention gel is applied to the roots and then wrapped in plastic to retain superior moisture for transit. They are placed in corogated cardboard shipping boxes for protection when shipped
Upon Receipt Of Your Plants
Upon receipt of your plants, unpack and unwrap the roots and mist with water. Plant within 24-48 hours. If you can not plant within this time frame, put your plants in a cool location (ex- basement, garage or cellar) and water the roots daily. Cover them back up with the plastic so they will not dry out until you can plant them. After planted, water every evening after the sun goes down for 5 days.
|Shipping Dates
Ferns are Beneficial for Landscaping
Ferns for shade
1.) How Ferns Are Beneficial in Landscaping
How Ferns Are Beneficial In Landscaping
If people are unable to see the forest for the trees, then imagine how easy it is to overlook the many benefits of the lowly fern in landscaping design. Ferns are such a common and essential part of the fabric of both field and forest; it's easy to see why this plant is dismissed. But various fern species provide environmental cover, act as food for both wild animals and humans, and even filter toxins from polluted soils, making them a critical "medic" in land restoration. And today, the "lowly" fern is occupying an increasingly important space in landscape planning on private land. Read on to see why.
Ferns Provide Excellent Ground Cover
1. There's A Fern For All Occasions
A few decades ago, there were less than ten species of Ferns for shade appropriate for backyard landscaping. Today, there's more than 150. And while even modified ferns remain nonflowering plants, their colors and shape more than make for lack of blossoms.
2. Ferns Command Respect
They do have animal predators and disease problems, but many fewer than other plant species.
3. Ferns Are Hardy And Don't Need Much Attention
A lack of or overabundance of moisture is possibly the single most significant deterrent to the thriving of a fern plant or colony. Working with professional landscapers is the best way to ensure the pairing of the right ferns with the right property.
4. Ferns Can Restore "Injured" Properties
Multiple studies have shown how ferns can filter toxins like heavy metals from land. Certain species of ferns such as marsh ferns provide excellent ground cover for very wet areas and can reduce damage from erosion.
Hardy as they are, fern species have particular requirements. This is why it's best to have property types assessed and recommendations made professionally before proceeding. With that in mind, here are a few of the more popular ferns for specific property types:
For Very Wet Areas
Native species like cinnamon ferns and sensitive ferns do well in wet areas, including thriving in bogs and at the edge of ponds.
For Dry Areas
Not all ferns need lots of water to thrive. There are fern species that can survive in desert conditions, though these plants are smaller and less showy. The gigantic wood ferns and lady ferns do very well in drier yards, however.
For Areas With Moderate Light And Moisture
Hart's tongue (needs extra lime), oak, and autumn ferns all do well in moderate light. They also require less moisture than some other species, provided that this moisture is received consistently.
Ferns for shade


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Austrian team Zele Racing have confirmed that Angolan-born Luis Sa Silva will make his Auto GP World Series debut in the season ending round at Estoril.
It will be the first time Sa Silva will compete in Auto GP, having raced primarily in the GP3 Series with Carlin for the past two seasons, but the 550bhp Auto GP car will be a big step up for him.
He will also need to adapt to the two compounds of Kumho tyres the series uses, whereas GP3 only uses one compound per weekend.
"I'm happy to drive such a powerful race car like the Auto GP one," insisted Sa Silva. "It will be a tough challenge but I'm happy that it will happen at Estoril. It's one of the tracks I know better as I turned lot of laps there. I also consider it my home circuit because I live in my second house not too far away from it".
Sa Silva will be Zele Racing's sixth driver of 2014, having seen Sergio Campana, Yoshitaka Kuroda, Christof von Grunigen, Tamas Pal Kiss and Samin Gomez all race for the team at some point during the season. The team were absent from the previous round at the Nurburgring.
The Estoril weekend marks the end of the 2014 Auto GP World Series season, with Kimiya Sato already being confirmed as champion.


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I go Nilla a Stall Snack on Saturday when she first injured herself. It's like this but without the attached ball. She didn't touch it once in 3 days so I gave it to Dijon who finished half of it in one day.
On Tuesday, I gave her a Jimmy's Hanging Ball. She was initially interested:
But she hasn't touched it since.
One thing she has maintained interest in the whole time is her treat ball. The barn owner said she hears her rolling it around the stall a lot during the day. She still has't really seemed to figure out how to get the treats out, since she's always trying to put her lips into the hole or pick it up by the hole, but she must be knocking some out by accident, since it does get emptier every time I check on her.
So I would highly recommend these treat balls if you have a bored horse. I put a mix of treats and hay pellets in it and that seems to work pretty well.
Does anyone else have any great boredom busters for horses? Nilla's allowed to use her paddock now, but she's still pretty bored without getting any work or pasture time.
Has anyone else read Next Town Over? It's a gorgeous web comic filled with pretty horses. One of the horses has a metal leg from an injury:
Nilla is going for the cyborg look herself:


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Top Charities
ACE's Top Charities work in ways that are most likely to produce the greatest gains for animals, actively evaluate and improve their programs, and have a demonstrated need for more funding. We promote them as our top giving opportunities, and as ideal examples of effective advocacy.
Albert Schweitzer Foundation
Albert Schweitzer Foundation (ASF) conducts corporate outreach campaigns and a variety of vegetarian outreach campaigns. They are one of the first animal charities conducting corporate outreach campaigns on behalf of fishes.
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Anima International
Anima International runs corporate campaigns, releases undercover investigations, and engages in online and offline ad campaigns and outreach. They work to build the capacity of the animal advocacy movement, including in relatively neglected regions.
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The Humane League
The Humane League runs a variety of programs that advocate for farmed animals, including corporate outreach, online veg advertising, and individual outreach. They continually seek to optimize the impact of their activities.
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The Good Food Institute
The Good Food Institute works in an underutilized but critically important area by promoting and developing competitive alternatives to animal-based meat, dairy, and eggs. They provide support in a variety of ways to cellular agriculture and cultured meat companies, including building relationships with distributors and retailers and driving R&D of the products.
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ACE's Standout Charities conduct strong work on behalf of animals in ways that could potentially affect large numbers of animals. We feel confident in the quality of their work, but for one reason or another we did not select them for a Top Charity recommendation. Standout Charities retain their status for two years, after which they are re-evaluated by ACE.
Compassion in World Farming USA (Compassion USA) is the U.S. branch of an international organization working to advance farmed animal welfare. They work to improve farmed animal welfare, primarily by changing corporate practices and policies through corporate campaigns and their Food Business program. Read Review.
Faunalytics works to support other animal organizations through research. They conduct independent research, work with client organizations on various research projects, and provide resources for individual advocates through the content library they host on their website. We think they are a particularly good giving opportunity for donors with a special interest in supporting research that can benefit the entire animal advocacy movement. Read Review.
Federation of Indian Animal Protection Organisations (FIAPO) works to build the capacity of the animal advocacy movement in India. Their primary focus is reducing the suffering of farmed animals. As farmed animal advocacy in India is relatively neglected, we believe that FIAPO's work has the potential to be highly effective. Read Review.
Sinergia Animal is a Brazilian organization that engages in corporate outreach with the goal of reducing animal suffering and decreasing the consumption of animal products. They focus especially on countries where major international animal advocacy organizations are not currently working, and aim to work as pioneers, paving the way for other organizations to expand their operations. We recommend Sinergia for donors who want to promote corporate reforms in relatively neglected countries. Read review.
Sociedade Vegetariana Brasileira (SVB) is a Brazilian organization that promotes plant-based diets by organizing veg fests, promoting Meatless Mondays, administering a vegan label, and working with restaurants to increase vegan offerings. They also work with health professionals to spread knowledge of the health benefits of a plant-based diet. We recommend SVB for donors who are interested in promoting plant-based diets in Brazil. Read review.
Considered Organizations
Since we began our charity evaluations in 2014, we have considered over 500 charities in total. For more details, see our complete list of Charity Reviews.


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Potters Bar business targeted by brazen burglar
PUBLISHED: 17:22 18 September 2018 | UPDATED: 17:22 18 September 2018
©2018 Danny Loo Photography - all rights reserved
The manager of a successful Potters Bar business has described his shock and disappointment after his High Street premises were burgled.
The culprit was caught on CCTV, first managing to somehow open the shutter and then fiddling around before opening the door.
Once inside the thief did not take long to quickly unplug two high-value computers, put them screen to screen, cover them in a sheet and walk out with them.
Colin Levy is the manager of Cre8business, which was targeted on the night of Monday, September 10.
He said: "It's quite a shock really.
"They unplugged two iMac computers very quickly."
The thief wore a baseball cap and left in the direction of Canada Life.
Mr Levy added: "He looked like he had an earpiece in so whether he had a driver waiting or an accomplice, I don't know.
"It was very organised.
"They targeted what they wanted.
"They came straight in and straight out.
"It's just the aftermath now because all the passwords have had to be changed.
"There's nothing on the hard drives.
"It's just the inconvenience."
The police forensics team was quick to visit the business premises and police have been in regular contact with Mr Levy, who is now offering a substantial reward for information that leads to the return of the two computers in their original condition.
He said: "It's an established business with a good reputation.
"I don't know if the person responsible came in as a customer to have a look.
"I don't know if this has happened anywhere else in the town."
Crebusiness opened in Potters Bar in April 2008 and since then has provided hundreds of local and national businesses with design, print and web services, as well as vehicle graphics.
A police spokeswoman said: "Police are investigating.
"Between 5.30pm on September 10 and 9.30am on September 11, two Apple Mac desktops were taken from Cre8 in the High Street.
"Any witnesses or anyone with information should contact Hertfordshire Constabulary on the non-emergency number 101, quoting crime reference 41/35715/18."
If you value what this story gives you, please consider supporting the Welwyn Hatfield Times. Click the link in the orange box above for details.


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The dialysis coordinator seemed to suggest that Peritoneal Dialysis (PD) seems to be the best option. I was shown a photo of a man who travels with PD & who leads a very active life. He seemed to look quite healthy. But that catheter tube that will stick out of one's abdomen is not ideal - it looks risky where infections are concerned. And the thought of having all that extra fluid in the abdominal cavity seems debilitating (for one thing, the missionary sexual position would be unviable because of abdominal pressure).
In the meantime, should I get the flu vaccine? I've never bothered with this thing before as I've not been prone to getting serious colds or flu. I've had a pretty good immune system. I am worried about any serious reactions to the flu vax - I seem to be becoming paranoid & anxious about the concept of being injected with chemicals lately.
But from everything I have learned - the pancreas tx is very risky to me because if the new pancreas fails - I can't go back to my native pancreas (which functions normally except for the deficient insulin cells). I wonder what your thoughts are on this UkrainianTracksuit?
My nephrologist is literally on the board of directors for my disease (PKD). She was the chief science adviser for five years
Athena,Okay, so, reading your post from a couple months ago now, I do have some thoughts for you to consider. I just don't frequent here very much.First of all, I too went to transplant eval recently. August. They were then, and still am today, shocked that I am not on dialysis. My egfr is around 17-18% and I only have one kidney!We are in similar places, except I am younger (46). I still lift weights (compound barbells, the real stuff) three times a week. Just today I squatted 3 sets of 195 lbs for 8 reps for my core lift. Then did 3 sets of 10 seated overhead presses at 75 lbs right after. Then I went to the easier stuff.I get to yoga as often as my schedule allows (twice a week, most weeks). I walk a fair bit still.My blood type is B+, which means I'm going to have a long, long wait for a cadaver kidney. Sucks.Anyway - I would suggest that, although I don't believe you should categorically refuse dialysis, I wouldn't necessarily let them force you onto it either. For the record, and I've told the doctors this - I'm going to fight to go without dialysis as long as I possibly can. I've read people functioning adequately at 8-10% eGFR. Everyone is different, and in my experience so far, very, VERY few people are as fit as I am. Additionally, there's not a single other health related problem of consequence, outside of the standard kidney related stuff. I just did an echo two weeks ago and crushed it. Got my BPM to a steady 160 and felt my knees would give out before my heart ever would. I was at like 15-16 on the borg scale at the very end. Also, I too have been recommended that an 'average' protein diet is fine. My nephrologist is literally on the board of directors for my disease (PKD). She was the chief science adviser for five years. I am 170 lbs and she wants me at 65-75g protein a day. Many days I'm under simply because the process of eating low salt, low potassium, and now lower phosphorus really makes eating healthy a struggle. Never mind maintaining muscle mass.I will add that as far as animal protein goes, my neph did suggest keeping it limited.You sound like a fighter. Keep fighting. It's hard, I have dark days and short periods of time. It sucks. However, I rebound. Because surrendering only expedites the problems. To hell with that.
When those who maintain a fitness routine are stricken by an illness, they are likely to have better outcomes than those with a similar illness who are out of shape


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Vibrio fischeri possesses two quorum-sensing systems, ain and lux, using acyl homoserine lactones as signaling molecules. We have demonstrated previously that the ain system activates luminescence gene expression at lower cell densities than those required for lux system activation and that both systems are essential for persistent colonization of the squid host, Euprymna scolopes. Here, we asked whether the relative contributions of the two systems are also important at different colonization stages. Inactivation of ain, but not lux, quorum-sensing genes delayed initiation of the symbiotic relationship. In addition, our data suggest that lux quorum sensing is not fully active in the early stages of colonization, implying that this system is not required until later in the symbiosis. The V. fischeri luxI mutant does not express detectable light levels in symbiosis yet initiates colonization as well as the wild type, suggesting that ain quorum sensing regulates colonization factors other than luminescence. We used a recently developed V. fischeri microarray to identify genes that are controlled by ain quorum sensing and could be responsible for the initiation defect. We found 30 differentially regulated genes, including the repression of a number of motility genes. Consistent with these data, ain quorum-sensing mutants displayed an altered motility behavior in vitro. Taken together, these data suggest that the sequential activation of these two quorum-sensing systems with increasing cell density allows the specific regulation of early colonization factors (e.g., motility) by ain quorum sensing, whereas late colonization factors (e.g., luminescence) are preferentially regulated by lux quorum sensing.
The association between the luminescent marine bacterium Vibrio fischeri and the Hawaiian bobtail squid, Euprymna scolopes, is a model system for the study of mutualistic bacterium-host interactions that has been successfully used to investigate the processes underlying the initiation, accommodation, and persistence of symbioses (33, 34). The association is initiated every generation, as newly hatched juvenile squids collect their bacterial symbionts from the surrounding seawater (30). Within the first hour after hatching, bacterial cells present in the ambient seawater trigger the secretion of mucus from the superficial epithelium of the squid light organ. V. fischeri cells attach to this mucus, forming dense aggregates. Between 3 and 6 h after hatching, the bacterial aggregates begin to move towards pores that lead into the interior of the light organ. After entering the pores, bacterial cells travel through ciliated ducts into the deep crypt regions of the light organ. Interestingly, this latter process is V. fischeri specific and requires flagellum-driven motility (30). Once inside the light organ, V. fischeri cells grow on nutrients supplied by the squid host and reach maximal colonization levels at approximately 24 h postinoculation (15, 35). Several genetic factors required for the different stages of this cooperative association have been identified; for example, (i) motility is essential for the initiation of the symbiosis (14, 26-28), (ii) the ability to acquire iron is important for the symbionts to maintain a normal level of colonization (16), (iii) defense against oxidative stress is necessary to efficiently colonize (44), (iv) luminescence expression is required for persistent colonization (41), and (v) the regulation of colonization gene expression is important at each of these stages (22, 28, 41, 45, 46).
The coordinated expression of colonization factors can be controlled by a process termed quorum sensing, which relies on the secretion of small signal molecules that accumulate in the surrounding environment. Once the bacterial population reaches a threshold cell density, the concentration of the quorum-sensing signal is sufficient to induce gene expression either directly by interacting with a transcriptional regulator or indirectly by activation of a signaling cascade. Thus, colonization factors are expressed only when they are beneficial to the bacterial cell (e.g., when they are associated with the host), avoiding the execution of costly processes in nonpermissive environments (11, 47). In gram-negative bacteria the most commonly used signals are either (i) acylated homoserine lactones (acyl-HSLs) synthesized by a LuxI or LuxM type of enzyme or (ii) autoinducer-2 (AI-2), a furanosyl borate diester synthesized by the LuxS enzyme (5, 47). While acyl-HSLs are generally species specific due to differences in the acyl side chain, the LuxS signal appears to be essentially ubiquitous; i.e., many gram-negative and gram-positive species produce and respond to this signal molecule (2).
Quorum sensing based on the lux system was first described in V. fischeri as the principal process regulating luminescence expression (7). The expression of the lux genes, luxICDABEG, in V. fischeri is regulated in a cell density-dependent fashion through the LuxI-directed synthesis of N-3-oxo-hexanoyl homoserine lactone (3-oxo-C6-HSL) and its subsequent binding to the transcriptional activator protein LuxR (8, 25). The LuxR-acyl-HSL complex interacts with the lux promoter and induces the transcription of the luxICDABEG locus (12). The V. fischeri lux system is required for both luminescence expression in vivo and persistence of the bacterial symbiont in the squid light organ (41). V. fischeri possesses a second quorum-sensing system based on AinS, an enzyme belonging to the LuxM type of acyl-HSL synthases, which synthesizes an N-octanoyl homoserine lactone signal (C8-HSL) (13). We previously proposed a model of luminescence gene regulation by the two quorum-sensing systems ain and lux (22) (Fig. 1). At low cell densities, luminescence gene regulation is repressed through LuxO (22), a negative modulator of the transcriptional regulator LitR (29), (Fig. 1). At the intermediate cell densities achieved in laboratory culture, the AinS-synthesized signal has two effects, i.e., induction of luminescence gene expression through direct interaction with LuxR and inactivation of LuxO (22); the second effect results in increased transcription of litR (29) (Fig. 1). LitR positively regulates transcription of V. fischeri luxR (9), thereby connecting the two quorum-sensing systems ain and lux and allowing the sequential induction of luminescence gene expression with increasing cell density (22) (Fig. 1). Inactivation of the V. fischeri ain system results in a luminescence defect and affects colonization competence (22). The V. fischeri LuxS signal AI-2 was shown to be involved in the regulation of luminescence expression and colonization competence as well; however, compared to that of ain and lux quorum sensing, the impact of AI-2 was quantitatively small (21).
We asked here whether the stepwise activation of the ain and lux quorum-sensing systems with increasing cell density is functional in the symbiosis. We found that (i) ain, but not lux, quorum sensing is required for efficient initiation of colonization; (ii) the lux quorum-sensing system is not maximally induced until the later stages of colonization; and (iii) ain quorum sensing differentially regulates several factors, such as flagellar motility, with potential importance for colonization initiation.
Bacterial strains and growth conditions.Strains and plasmids used in this study are listed in Table 1. V. fischeri strains were grown at 28°C either in a seawater-based nutrient medium (SWT) (3) or in Luria-Bertani salt medium (LBS) (14). E. coli strains were grown at 37°C in Luria-Bertani medium (36). Media were solidified with 1.5% (wt/vol) agar as needed. Antibiotics were added to the media at the following concentrations when appropriate: chloramphenicol (Cam), 2.5 μg/ml for V. fischeri and 20 μg/ml for Escherichia coli; erythromycin, 5 μg/ml for V. fischeri and 150 μg/ml for E. coli; and kanamycin (Kan), 100 μg/ml for V. fischeri and E. coli. 3-Oxo-hexanoyl-l-homoserine lactone (3-oxo-C6-HSL) was obtained from Sigma-Aldrich Corp. (St. Louis, MO); octanoyl-l-homoserine lactone (C8-HSL) was obtained from Aurora Biosciences (Coralville, IA). Other medium reagents were purchased from Difco Laboratories (Sparks, MD) and Sigma-Aldrich.
Colonization assays.Colonization levels were determined as the number of CFU per squid light organ present at 12 h postinoculation (34). For each experiment, 12 to 20 newly hatched squids were placed into 50 ml of filter-sterilized seawater containing about 3,000 CFU of the indicated strain per ml. Another group of animals was placed into filter-sterilized seawater without added bacteria. After incubation for 12 h in the dark to prevent the usual expulsion of bacteria at sunrise (15, 19), the animals were homogenized, and the homogenate was diluted and spread onto SWT agar. The colony number was counted after overnight incubation, and the mean number of CFU per squid was calculated.
To determine the relative competitive index of the mutant strains, 10 to 15 newly hatched squids were placed into 50 ml of filter-sterilized seawater containing approximately 4,000 CFU of each of two competing strains and were incubated for 12 h. An aliquot of the inoculated seawater was spread onto LBS agar to determine the number and exact ratio of the two strains in the inoculum. At 48 h postinoculation, squids were homogenized and a dilution of the homogenate spread onto LBS agar. Approximately 100 CFU from the inoculum, as well as from each of the homogenates, was patched onto both antibiotic-containing and antibiotic-free LBS agar to determine the ratio of V. fischeri wild-type (Cam and Kan sensitive) to luxO mutant (Kan resistant) or ainS mutant (Cam resistant) cells. The relative competitive index was calculated as the ratio of the two strains in the symbiosis divided by the ratio in the inoculum.
To observe aggregation behavior, newly hatched squids were incubated with an inoculum of 500,000 CFU per ml of either wild-type V. fischeri or the ainS mutant, each carrying pKV112, a green fluorescent protein expression plasmid (Table 1). Every 30 min, two squids per treatment were dissected and the formation of aggregates was observed using fluorescence microscopy as previously described (30).
Mutant construction.Genetic techniques (DNA extraction, PCR, ligation, conjugation, and DNA sequencing) were carried out as previously described (22).
The LuxO(D47E) mutant was constructed using the QuikChange site-directed mutagenesis kit according to the manufacturer's protocol (Stratagene Inc., La Jolla, CA). Plasmid pCL145 (Table 1), carrying the intact luxO gene in the allelic exchange vector pEVS79 (Table 1), served as a template to create the site-directed mutation. The asparagine GAT codon in position 47 of the LuxO protein was changed into a glutamine GAG codon by introducing a T→G point mutation. The resulting plasmid, pCL150, was transferred into the V. fischeri luxO mutant CL42 by triparental mating, and double recombinants that had lost kanamycin resistance were selected as previously described (39), generating the V. fischeri LuxO(D47E) mutant strain CL59. The introduction of the point mutation into the genome was confirmed by sequence analysis of the mutant's luxO gene. Luminescence and growth characteristics were determined as previously described (22).
Microarray comparison.Wild-type V. fischeri and the ainS, luxO, and LuxO(D47E) mutants were grown in SWT medium to an optical density (OD) at 600 nm of 2.5, a point at which V. fischeri cells still grow exponentially and, based on luminescence expression data (22), ain quorum sensing is maximally induced. RNA extractions were performed as previously described (37). cDNA synthesis was carried out using 10 to 12 μg of purified RNA, 15 μg random decamer primers (IDT, Coralville, IA), a deoxynucleoside triphosphate mixture (Amersham Biosciences, Piscataway, NJ) containing a 3:2 ratio of aminoallyl-dUTP (Sigma-Aldrich) to dTTP, and 500 units of Superscript II reverse transcriptase (Invitrogen Life Technologies, Carlsbad, CA). RNA was removed by alkaline hydrolysis, and cDNA was purified and concentrated using Microcon-30 filters (Millipore Corp., Billerica, MA). Labeling of cDNA was carried out using monoreactive Cy3 and Cy5 dyes according to the manufacturer's protocol (Amersham Biosciences), and labeled cDNA was desalted with a PCR purification kit (QIAGEN Inc., Valencia, CA). Nucleotide and label dye concentrations were determined by measuring absorption at 260 nm (nucleotide), 550 nm (Cy3), and 650 nm (Cy5). Microarray glass slides spotted three times with PCR products representing approximately 95% of all V. fischeri open reading frames (Integrated Genomics Inc., Chicago, IL) were incubated for 45 min at 42°C in prehybridization solution (5× SSC [1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate] [Sigma-Aldrich], 0.001% sodium dodecyl sulfate [SDS], 0.5% bovine serum albumin). The two differentially labeled samples were mixed at a Cy3:Cy5 dye ratio of approximately 1:1 and concentrated to a final volume of 10 μl using Microcon-30 columns (Millipore Corp.). To each sample, 3.5 μl of 1-mg/ml salmon sperm DNA, 3.5 μl of 1-mg/ml yeast tRNA, and 18 μl PerfectHyb Plus hybridization buffer (Sigma Chemical Co.) were added, and the mixture was denatured at 99°C for 1.5 min. Each sample was added to the microarray glass slides and hybridized for 14 to 18 h at 42°C. The slides were washed at approximately 24°C for 5 min each in (i) 1× SSC, 0.001% SDS; (ii) 0.1× SSC, 0.001% SDS; (iii) 0.001% SDS; and (iv) water. After drying, fluorescence intensities were determined using a 428 Array Scanner (Affymetrix Inc., Santa Clara, CA). For each hybridization spot, background intensities were subtracted from spot intensities. The data were then log transformed and normalized by equalizing 25% and 75% tiles. Ratios of the normalized Cy3 and Cy5 dye intensities were determined for each spot and averaged for each microarray. For statistical analysis, a two-sided, paired Student t test was performed.
Transcriptional activity of flaA.Plasmid pDM104-6 (Table 1) was introduced into V. fischeri wild-type and ainS mutant strains by triparental mating as previously described (39). Both strains were cultured in SWT plus erythromycin, and flaA transcriptional activity was estimated by assaying beta-galactosidase activity in six samples from two independent cultures at different time points during growth, using a standard method (36).
Motility assays.Motility and chemotaxis behaviors of V. fischeri were determined by growing wild-type and mutant strains in the appropriate liquid medium to an optical density of approximately 0.4. Three microliters of the culture was spotted on the surface of a fresh 0.25% agar medium plate, and the relative diameters of the colony and the expanding rings of migrating cells were determined after 6 to 24 h of incubation at 25°C. Assays were carried out on (i) SWT medium or (ii) defined medium (10 mM NH4Cl, 0.33 mM K2HPO4, 300 mM NaCl, 50 mM MgSO4, 10 mM CaCl2, 10 mM KCl, 0.01 mM FeSO4, and 50 mM Tris-HCl [pH 7.4]) containing either 0.3% (wt/vol) Casamino Acids (CAA) or 0.5 g/liter N-acetylglucosamine (NAG) as a carbon source.
Colonization initiation by V. fischeri quorum-sensing mutants.To determine whether any of the V. fischeri quorum-sensing systems is important for the initial stages of light-organ colonization, we determined the number of CFU in the organ 12 h after inoculation with either wild-type V. fischeri or the luxI, ainS, or luxS mutant (Table 2). At this early point in colonization, the number of CFU in squid colonized with the luxI mutant was indistinguishable from wild-type levels (Table 2). The fact that luxI mutant-colonized animals do not produce detectable luminescence (i.e., <1% of the wild-type level) (22) suggests that even a severe defect in light production does not affect colonization efficiency during the initial stages of colonization. In contrast, the ainS mutant, while expressing low (only 10% of wild-type) luminescence levels (22), was able to colonize the light organ only to <50% of the wild-type number of bacterial cells. Thus, although light emission was affected by a mutation in ainS, this gene probably regulates additional factors beyond luminescence that are important for colonization initiation. Squid colonized by the luxS mutant displayed colonization levels indistinguishable from those of wild-type-colonized animals at 12 h postinoculation (Table 2), indicating that luxS is not required for normal initiation of colonization. A secondary mutation in either luxI or luxS did not enhance the colonization defect of the ainS mutant (Table 2), suggesting the absence of a synergistic effect of these mutations in colonization initiation.
One of the early events in colonization initiation is the formation of bacterial aggregates outside the squid light organ (30). Because a defect in this activity could explain the observed initiation phenotype, we examined aggregation behavior of the ainS mutant in comparison to wild-type V. fischeri. However, we could not detect any difference between the two strains, either in the timing of aggregate formation or in the size of the bacterial aggregate formed (data not shown). These results suggest that the ainS mutant is not defective in attaching to the squid mucus and that the colonization process(es) regulated by ainS becomes important after bacterial aggregation.
Regulation cascade of the initiation defect.In our previous study (22) we demonstrated that the ainS mutant displays a persistence defect; i.e., the number of CFU is similar to that of the wild type at 24 h after colonization (the earliest time point examined) but becomes significantly lower by 48 and 72 h. This persistence defect could be reversed by introducing a luxO mutation into the ainS mutant background, suggesting that AinS regulates gene expression through the inactivation of LuxO (22). LuxO, in turn, down-regulates transcription of litR, a gene encoding a positive regulator of luxR (9, 29). To investigate whether the colonization factors underlying the initiation defect of the ainS mutant are regulated through this AinS-LuxO-LitR pathway (Fig. 1), we compared light-organ colonization levels at 12 h postinoculation with either the V. fischeri wild-type, luxO, ainS-luxO, litR, or luxR strain (Table 2).
Previous work (22) has shown that a luxO mutation does not interfere with the ability of V. fischeri to persist in the squid light organ after 24 h of colonization. However, the results presented here demonstrate that luxO, like ainS and litR but not luxR, is required for a normal level of colonization by 12 h (Table 2). Furthermore, the luxO mutant displayed a competition defect that was comparable to that of the ainS mutant. Specifically, when juvenile squids were colonized with a 1:1 mixed inoculum of either luxO mutant and wild-type cells or ainS mutant and wild-type cells, the average relative competitive index of both mutants was 0.4 (data not shown). Thus, mutations in both positive (AinS and LitR) and negative (LuxO) regulators in the pathway (Fig. 1) resulted in a similar defect in the level of colonization at 12 h (Table 2), which might also lead to a competitive defect. One interpretation of this surprising outcome is that, rather than a simple on-off switch, the AinS-LuxO-LitR pathway serves to maintain the colonization factor(s) it regulates at a proper level of expression. Because inactivation of luxR had no effect on the initiation of symbiosis (Table 2), the lux system is apparently not important for early stages of colonization, and thus the initiation defect of the ainS mutant does not operate through LuxR (Fig. 1).
Growth, luminescence, and colonization phenotypes of the V. fischeri LuxO(D47E) mutant.While inactivation of V. fischeri ainS results in both initiation and persistence defects, a mutation in litR, operating downstream of ainS, affected only colonization initiation (Table 2) (9, 22), suggesting that the effect of ainS on colonization initiation may be distinct from that on colonization persistence. We therefore hypothesized that the AinS-LuxO-LitR cascade might directly activate genes important for colonization initiation, while the absence of ain-dependent induction of lux quorum sensing results in a defect in colonization persistence (Fig. 1).
To investigate this possibility further, we constructed a site-directed LuxO(D47E) mutant encoding an aspartate-47-to-glutamate substitution that has been shown to mimic the phosphorylated, and thus active (i.e., repressive), state of the Vibrio harveyi LuxO protein (10). According to the model of quorum sensing (Fig. 1), we predicted that this mutation would simulate the effects of an ainS mutation on the LuxO-LitR regulatory cascade but would allow the induction of lux quorum sensing (22). When we compared the growth, luminescence, and colonization phenotypes of the LuxO(D47E) mutant with those of the ainS, litR, and luxO mutants (Table 3), we found that their behavior was consistent with our hypothesis. Specifically, the growth yield of the LuxO(D47E) mutant in culture was reduced, while its doubling time was indistinguishable from that of the wild-type strain. However, the growth yield defect of the LuxO(D47E) mutant was smaller than those of the ainS and litR mutants, suggesting that LuxO(D47E) might not repress downstream genes to the same extent as the native phosphorylated LuxO protein. As expected, the growth defect could not be eliminated by exogenous addition of C8-HSL (data not shown), because the mutated LuxO(D47E) protein only mimics the phosphorylated LuxO state and cannot be dephosphorylated (and thus deactivated) by ain signaling. Luminescence of the LuxO(D47E) and litR mutants in culture was decreased compared to wild-type levels but was not eliminated (Table 3) (9). This result is consistent with the model of regulation (Fig. 1) in which AinS has two effects on luminescence regulation: the inactivation of LuxO repression and the activation of the lux operon through a C8-HSL-LuxR interaction (22). Thus, inactivation of AinS is expected to result in a more severe luminescence defect than a constitutively active LuxO protein or a mutation in litR. The luminescence levels of the LuxO(D47E) and litR mutants did not change in response to exogenous C8-HSL addition, implying that both mutant strains synthesize sufficient amounts of C8-HSL to induce luminescence through C8-HSL-LuxR interaction. When 3-oxo-C6-HSL was provided, luminescence reached nearly wild-type levels, indicative of proper lux quorum-sensing induction. As previously shown, the ainS mutant displayed essentially opposite effects, i.e., an increase in luminescence to wild-type levels when C8-HSL was added but a reduced response to 3-oxo-C6-HSL (22) (Table 3). This effect may be due to the absence of C8-HSL-LuxR interaction, which is necessary for proper lux quorum-sensing induction (22).
Finally, while the colonization levels of the LuxO(D47E), litR, and ainS mutants were comparable at 12 h postinoculation, only the ainS mutant displayed a persistence defect at 72 h (Table 3). Thus, the ainS mutant is compromised in both the initiation and persistence of symbiosis; however, the underlying causes of these two colonization defects appear to be distinct.
Onset of lux quorum sensing.Having established that lux quorum sensing does not affect the ability of V. fischeri to initiate colonization, we asked whether this system is fully active during the early stages of colonization. Because induction of the lux system is required for light expression in the host (i.e., luxI mutant-colonized animals are dark [22]), lux activity can be easily monitored by measuring symbiotic light production. In culture, where the lux system is inactive, the exogenous addition of saturating concentrations of the LuxI-synthesized 3-oxo-C6-HSL stimulates light expression more than 1,000-fold (22). We used a similar approach to determine the level of lux activity during the initiation of symbiosis by comparing luminescence expression by wild-type-colonized juvenile squid exposed to seawater containing (i) no addition, (ii) 120 nM 3-oxo-C6-HSL, or (iii) 1,200 nM 3-oxo-C6-HSL during the first 24 h of colonization (Fig. 2A). To minimize any possible effects of 3-oxo-C6-HSL on colonization phenotypes other than luminescence expression, juvenile squids were incubated with wild-type V. fischeri cells for 6 h before being transferred into seawater containing 3-oxo-C6-HSL. Almost immediately after exposure to 3-oxo-C6-HSL, and 3 h earlier than the no-addition control, squid became detectably luminous (Fig. 2A). This positive effect of 3-oxo-C6-HSL on luminescence expression continued until approximately 12 h postinoculation, after which luminescence levels were indistinguishable between exposed and control animals (Fig. 2A). In contrast, exposure of wild-type-colonized squid to 1,200 nM of the AinS-synthesized signal C8-HSL had no significant effect on the development of luminescence expression (data not shown), suggesting that this signal is produced in saturating concentrations by 6 h postinoculation. Because acyl-HSLs are susceptible to hydrolysis, it was a concern that the absence of any positive effect on luminescence at time points beyond 12 h was due to degradation of the compound. However, because luminescence expression by animals exposed to 10 times the saturating concentration (1,200 nM 3-oxo-C6-HSL) was indistinguishable from that by animals exposed to 120 nM 3-oxo-C6-HSL (Fig. 2A), it seemed unlikely that degradation of 3-oxo-C6-HSL affected the outcome of this experiment.
Finally, we tested our hypothesis that LuxR, a target of LitR induction (Fig. 1), is limiting during the early stages of colonization. When juvenile squids were colonized with wild-type V. fischeri overexpressing the luxR gene in trans, luminescence levels were higher than those of animals colonized with V. fischeri carrying the vector control in trans (Fig. 2B). Again, the differences in the levels of light expression were observed only during the initiation of colonization. These data suggest that, because of the limited expression of LuxR, the lux system is not maximally induced until approximately 12 h postinoculation, coincident with the time at which the squid light-organ crypts become densely colonized with V. fischeri cells (30).
Microarray comparison of wild-type V. fischeri and quorum-sensing mutants.In an effort to identify factors regulated by V. fischeri ain quorum sensing that could be responsible for the observed colonization defects, we performed three independent microarray experiments using culture-grown cells. Two microarray experiments compared the transcriptomes of the ainS mutant and the wild type. To ensure that the regulation was ain quorum-sensing specific and not a result of gene regulation by the C8-HSL-LuxR interaction (Fig. 1), a third experiment compared the transcriptomes of the LuxO(D47E) and luxO mutants. We used the two selection criteria of (i) greater than twofold differential regulation in each of the three experiments and (ii) a P value of <0.05 to identify genes that were differentially regulated by ain quorum sensing (Table 4).
The most striking finding of the microarray study was that genes encoding several flagellins and flagellar basal-body proteins, appeared to be negatively regulated by the V. fischeri ain quorum-sensing system (Table 4). In addition, one of the 31 putative methyl-accepting chemotaxis protein genes (http://www.ergo-light.com/ERGO/ ) appeared to be differentially repressed by ainS, suggesting a possible role of quorum sensing in the regulation of a behavioral response to a specific environmental condition (Table 4). Finally, the gene for the V. fischeri FlgM homolog, a putative sigma-28 antagonist, appeared to be down-regulated as well, indicating that the repression of sigma-28-dependent motility genes by ain quorum sensing is unlikely to function through FlgM (Table 4). The majority of the V. fischeri motility genes are located in a distinct chromosomal region (VF1826 to VF1881 [http://www.ergo-light.com/ERGO/ ]). When we lowered the selection criteria somewhat to include genes that were on average greater than twofold differentially regulated (yet still with a P value of <5%), we found an additional six motility genes that were down-regulated by ain quorum sensing (Table 4).
We confirmed the microarray data for one of these genes (flaA) by comparing the beta-galactosidase activity levels of a flaA::lacZ fusion carried in trans by both wild-type V. fischeri and the ainS mutant. The beta-galactosidase activity was on average 3.3 times higher in the ainS mutant than in wild-type V. fischeri (data not shown), supporting a role of ainS in the expression of this important flagellin (26).
The genes for several regulatory proteins also appeared differentially regulated by ain quorum sensing (Table 4), suggesting that the V. fischeri quorum-sensing regulon is connected to other regulatory circuits. For instance, the previously proposed positive regulation of V. fischeri litR by AinS (22) was confirmed (Table 4). While the V. fischeri ainS mutant does not express detectable luminescence levels in culture and the specific luciferase activity is only 10% of that of the wild type (22), we could not detect differential luminescence (luxCDABE) or regulatory (luxR) gene expression by ain quorum sensing. We believe that this result may reflect how luciferase specific activity increases dramatically around an OD of approximately 1.0 and stays constant thereafter (22). Thus, at the OD of 2.5 at which the sample was taken for microarray analysis, positive regulation of lux transcription by ain quorum sensing may have already ceased.
One interesting regulatory locus contains the genes VFA1013 to VFA1017, some of which were positively regulated by ain quorum sensing (Table 4). The proteins encoded by both VFA1014/VFA1013 and VFA1016/VFA1017 have regions homologous to protein kinases and response regulator pairs, respectively (http://www.ergo-light.com/ERGO/ ). Located between these putative two-component regulatory systems is another gene (VFA1015) that was positively regulated by ain quorum sensing (Table 4). This gene is most closely related to the stationary-phase sigma factor gene V. fischeri rpoS (VF2067) (http://www.ergo-light.com/ERGO/ ). The functions of the genes in this regulatory locus remain to be determined; nevertheless, the ainS regulation of VFA1015, together with the growth yield defect of the ainS mutant (Table 3), led us to the hypothesis that the genes in the VFA1013-to-VFA1017 locus may encode important sensory and response systems used by V. fischeri to switch from the planktonic to the symbiotic lifestyle.
Motility phenotype of V. fischeri quorum-sensing mutants.Because the microarray results indicated that flagellin and flagellar basal body genes are regulated by the ain system, we assessed the motility behaviors of several of the quorum-sensing mutants. As we reported previously (22), the swimming behaviors of the ainS and the luxI mutants on rich SWT medium containing either 0.4% or 0.7% agar were indistinguishable from that of the wild type. However, at a lower agar concentration (0.25%) on both SWT and a minimal salts medium supplemented with either CAA or NAG, differences in motility behavior between wild-type and mutant strains became apparent (Table 5).
In general the effects of quorum-sensing gene mutations on motility were more pronounced in the defined media (Table 5), suggesting that nutrient availability affects V. fischeri motility by an unknown mechanism. While the ainS mutant displayed a hypermotile phenotype in all media, the luxO mutant produced slightly smaller migration ring diameters in SWT, and no migration could be observed in minimal medium containing either CAA or NAG (Table 5). The migration patterns of the ainS-luxO mutant in each medium were indistinguishable from those of the luxO mutant, suggesting that luxO has a dominant effect on the motility phenotype. The luxS mutant migrated slightly faster than the wild type in SWT, but the migration was not affected in minimal medium (Table 5). Furthermore, the introduction of the luxS mutation into the ainS mutant did not noticeably enhance the hypermotile phenotype of the ainS mutant (Table 5), suggesting the absence of a synergistic effect. The litR mutant displayed essentially the same phenotype as the ainS mutant (Table 5), consistent with the gene regulation model (Fig. 1) in which litR functions downstream of ainS (22). Neither mutation of luxI nor mutation of luxR resulted in an altered motility behavior under the conditions tested (Table 5). All of the strains grew with identical doubling times in each of the media, and only the hypermotile ainS, ainS luxS, and litR strains (22) (Table 3) displayed a growth yield defect (data not shown), indicating that the observed phenotypes are not due to a metabolic defect.
In this study we investigated the impact of the V. fischeri quorum-sensing systems ain and lux on the early stages of colonization. Our data demonstrate that (i) ain (but not lux) quorum sensing is required for the normal initiation of colonization, (ii) the lux system is not fully active during initiation of colonization, and (iii) the ain system regulates not only the lux system but also several putative early colonization genes, including those controlling motility. Taken together, these data demonstrate for the first time the differential activation of two bacterial quorum-sensing systems at distinct stages of a beneficial host tissue colonization.
A model of colonization factor regulation by V. fischeri quorum sensing.Many gram-negative bacteria possess multiple quorum-sensing systems (47), a phenomenon that has led to an examination of how these systems may function together. Our results show that in the case of V. fischeri two systems are arranged to allow the induction or repression of colonization factors important during specific temporal phases of the symbiotic relationship. The fact that the ain system is functional and essential at early stages of colonization, whereas the lux system is neither fully induced nor required at this time (Fig. 2), suggests that the ain system is operative at a lower threshold cell density than the lux system and provides evidence for a stepwise pattern in their control during the onset of symbiosis. The sequential nature of quorum-sensing induction is further supported by our previous finding that in symbiotic strains of V. fischeri the effect of the ain system on luminescence is apparent at the relatively moderate bacterial concentrations occurring in culture (108 to 109 cells ml−1), whereas the lux system is the predominant inducer of luminescence expression at the very high cell densities (>1010 cells ml−1) achieved only within the squid light organ (22).
We proposed previously a model by which luminescence is regulated by ain and lux (22). The results of this present study expand this model to include the regulation of non-lux genes at specific phases in the colonization of E. scolopes by V. fischeri. At the very low cell densities found in seawater (<104 cells/ml), the absence of ain quorum sensing allows repression of colonization gene expression through the transcriptional regulator protein LuxO (Fig. 1). When V. fischeri initiates colonization of its host by aggregating and then migrating through the squid light-organ ducts (30), the cell density increases to the threshold cell density of ain quorum sensing. This threshold level is approximately 108 cells/ml, based on both ain-dependent luminescence induction and C8-HSL signal production in culture (21, 22), and correlates with the cell densities occurring during the initiation process (35). At this stage the AinS signal, C8-HSL, inactivates LuxO, allowing the appropriate level of expression of early colonization genes through LitR (Fig. 1). This model is consistent with two observations: the ainS, LuxO(D47E), and litR mutants are similarly defective in colonization initiation, and a double ainS luxO mutant displays an initiation defect comparable to those of the single ainS and luxO mutants (Table 2), suggesting that their effects are modulated through a common pathway. While our evidence supports the notion that both luminescence genes (22, 29) and motility genes (Tables 4 and 5) are regulated through the AinS-LuxO-LitR pathway (Fig. 1), it remains possible that LuxO regulates some colonization factors independently of LitR.
We have previously demonstrated that ain quorum sensing is required for appropriate expression of the lux quorum-sensing system (22). Thus, inactivation of ainS produces two effects in vivo: an initiation defect resulting from the loss of regulation of early colonization factors (Table 2) and a persistence defect caused by the absence of appropriate lux quorum-sensing induction (22). As predicted, both the LuxO(D47E) and the litR mutants display an initiation defect (Table 2) but not a persistence defect (Table 3) (9).
Finally, once V. fischeri has densely colonized (>1010 cells/ml) the deeper crypt regions of the squid light organ, and after initial induction by the ain system, lux quorum sensing induces expression of late colonization genes, such as those encoding luminescence (Fig. 1). A previous study demonstrated that mutations in either luxI, luxR, or luxA (luxA encodes one of the subunits of luciferase), produced similar defects in colonization persistence (41), suggesting that luminescence might be the only colonization factor regulated by the lux quorum-sensing system. However, a separate study of the fish light-organ symbiont V. fischeri MJ1 showed that lux quorum sensing regulated the expression of five non-lux proteins (4). Inactivation of one of the corresponding genes, qsrP, in the squid light-organ symbiont V. fischeri ESR1 resulted in a competitive defect, suggesting that at least this lux quorum-sensing-regulated protein is a colonization factor.
Quorum sensing affects V. fischeri motility.When planktonic V. fischeri cells colonize juvenile E. scolopes, both the animal host and the bacterial symbiont undergo dramatic developmental changes (24). Two of the most apparent phenotypes of symbiotic V. fischeri cells are luminescence induction and an apparent loss of flagella (35). The ability to express luminescence during colonization is principally regulated by lux quorum sensing (42). Based on the data presented here (Tables 4 and 5), we hypothesize that, while flagellated planktonic V. fischeri initiate the colonization process, as they reach the deeper crypt tissues where proliferation occurs, flagellar elaboration is repressed as a consequence of ainS signaling.
There are several lines of evidence that the altered (either decreased or increased) motility behavior of mutants in the ain quorum-sensing pathway is the cause of the colonization initiation defect (Fig. 1; Table 2). Proper motility behavior has been shown previously to be essential for colonization initiation; i.e., both nonmotile and hypermotile mutants are significantly impaired in establishing a light-organ colonization (14, 27). However, at later stages of colonization the majority of bacterial cells have lost their flagella, implying that proper swimming motility is not required for maintenance of the symbiosis (35). The results presented here are consistent with this paradigm: both the hypermotile ainS and litR mutants and the motility-deficient luxO mutants (Table 5) display a delay in colonization initiation (Table 2). The presence of this delay in mutants predicted to have either higher (luxO) or lower (ainS or litR) expression of LitR suggests that targets of this regulator, like motility genes, must be carefully modulated.
Whole-genome transcription analysis showed that ain quorum sensing regulates both flagellar basal body and flagellin genes (Table 4). Motility gene regulation follows a hierarchical regulation cascade allowing the assembly of the flagellar apparatus in a sequential manner. The different classes of motility genes include early genes encoding regulatory proteins that control the entire motility regulon, middle genes encoding structural components of the hook-basal body and the type III secretion apparatus, and late genes encoding the flagellar filament, chemotaxis, and motor force generator proteins (1). The motility regulatory cascade in V. fischeri has not been completely elucidated; however, the analysis of regulatory proteins and the promoter regions of motility genes shows similarities to the regulatory cascade of Vibrio cholerae (26, 28, 31, 48). For example, inactivation of the early sigma-54-dependent master regulator (flrA), as well as sigma-54 (rpoN) itself, abolishes motility, suggesting that, as in V. cholerae, flrA is the master regulator of the V. fischeri motility regulon (28, 31, 48). Because ain quorum sensing regulates both sigma-54-dependent middle genes (i.e., flgBCDEM, flhF, fliEL, and flaA) (28) and sigma-28-dependent late genes (i.e., flaBCDEF) (26), it is not clear at which regulatory level quorum sensing affects motility gene regulation. Interestingly, while V. cholerae quorum sensing also negatively regulates motility, only chemotaxis genes were shown to be differentially regulated (49), suggesting that the regulatory level at which quorum sensing affects the motility regulon is different in these two Vibrio species.
ain quorum-sensing regulon.While our findings are consistent with the hypothesis that the altered motility behavior of the ain quorum-sensing mutants is, by itself, sufficient to explain their inability to efficiently initiate colonization, it remains possible that AinS and LuxO also regulate other, perhaps disparate, initiation factors. Our transcriptome analysis revealed several additional genes that are regulated by ain quorum sensing (Table 4) and might play a role in colonization initiation. For instance, V. fischeri may adapt to growth conditions in the light-organ crypts by using ain quorum sensing to regulate factors involved in nutrient uptake or metabolism (Table 4). Furthermore, the differential regulation of these genes provides a possible explanation for the growth yield defect exhibited by the ain quorum-sensing mutants in vitro (22) (Table 3).
Another result of ain quorum sensing appears to be the induction of genes in a putative exopolysaccharide (EPS) locus (Table 4). This locus contains many genes homologous to the Escherichia coli wzy locus, a system involved in O-antigen biosynthesis (VF0151 to VF0201 [http://www.ergo-light.com/ERGO/ ]). The consequences of EPS production in bacterium-host associations are diverse, but they can include enhanced adherence to tissue surfaces and/or protection against the challenges of complement-mediated phagocytosis and killing by the innate immune system (32). While the role(s) of EPS in this association remains to be determined, both of these challenges are faced by symbiotic V. fischeri (42). Mutations in homologs of LuxO or LitR in other Vibrio species affect colony morphology and biofilm formation, which in some cases correlates to changes in EPS production (6, 20, 23, 38, 49). Accordingly, the V. fischeri litR mutant was reported to display a more translucent colony morphology and to produce more biofilm than the wild type (9; P. Fidopiastis, personal communication). Not surprisingly, we found that the V. fischeri ainS mutant displayed similar phenotypes (data not shown); however, it remains to be determined whether these phenotypes are related to LPS and/or EPS production.
Our analysis indicates that ain quorum sensing has an effect on the transcription of a number of genes, including some unique ones (VF1355 and VF2255) with unknown functions (Table 4). While the functions of others can be inferred, it is not yet clear whether their regulation is direct or is an indirect effect of a multistep signaling pathway involving other transcriptional factors. In any case, the discovery of these regulated genes not only has given us insight into the way in which ainS signaling controls both early and late stages of symbiosis but also has provided candidate genes that may encode novel colonization determinants.
We thank Amy Schaefer for developing the protocol used in the microarray study and for help with the experiment, David Goode for performing normalization of the microarray data sets and advice on statistical analysis, and K. Visick for kindly providing plasmids. We are grateful to Debbie Millikan and Amy Schaefer for critically reading the manuscript. V. fischeri ES114 genomic sequence information was made available by Integrated Genomics Inc., Chicago, Ill.
This work was supported by National Institutes of Health grant RR12294 to E. G. Ruby and M. McFall-Ngai, by National Science Foundation grant IBN0211673 to M. McFall-Ngai and E. G. Ruby, and by a W. M. Keck Foundation grant to E. P. Greenberg, E. G. Ruby, M. McFall-Ngai, and others.
- Received 14 December 2004.
- Accepted 2 March 2005.
- Copyright © 2005 American Society for Microbiology


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Just who is this woman on the top of this page? It's Dale, head of the grandaddy of Atlanta food fests, Taste of Atlanta. She'd really love to see you this weekend. Taste of Atlanta has bounced around the city in year's past and has seemed to have found it's groove in Midtown at Technology Square. It's been blessed with great weather the past several years and it looks like we're in for a repeat performance this year.
I guess is the point where I tell you that hundreds of restaurants will be offering tastes of deliciousness, there will be chef demos, wine tastings, booze in the VIP tent etc ( I guess I just did?. I'd love to elaborate, but I'm actually running late for lunch with my man Brad, Mr. Thirsty South.
One of the highlights last year for me was a session of Inside the Food Studio, which was a ticketed event last year and now is open to the public on a first come, first served basis. I hate regurgitating press releases, but I'll do so on this occasion. Please forgive me y'all.
The 2011 Inside the Food Studio session highlights include:
- · A Knack for Global Snacks, presented by John Kessler: The seasoned AJC critic guides this journey across the world as Atlanta's own Top Chef contestant Hector Santiago of Pura Vida joins Guy Wong of Miso Izakaya to discuss and sample the international snacks the chefs grew up loving.
- · Simply Southern Thanksgiving: With the turkey-centric feast looming, guests delve into the dishes and traditions of a southern feast and get inspired with modern twists on all their hearty favorites. Chef Ford Fry of JCT. Kitchen and Drew Belline of No. 246 dish on how to dazzle at this year's Thanksgiving table.
- · Tongue-in-Cheek Tacos and Tequila: Revelers can get in on the fiesta with this spicy session. Guests get the scoop on the history and culture behind this idyllic duo that is pushing the boundaries of Atlanta's dining scene.
- · Cake Crusade: Some of Atlanta's top pastry chefs go head-to-head for a decorating throw down. When the secret theme is revealed, the sweets savants craft their custom creations. For some sugary justice, the audience votes on who takes the cake
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text: Cryptid Con is Kentucky's LEGENDary Bigfoot and Paranormal convention. Come meet media celebrities, authors, lecturers, vendors and other cryptozoology and legend enthusiasts for an amazing weekend event! We are bringing you a monstrous weekend of fun that you won't find anywhere else in the Bluegrass. Meet celebrity guests. Learn from experienced researchers and authors. HearRead More →


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We are an independent Audiology clinic so you can be assured our recommendations are based on the brand we feel best suits you. We are pleased to offer a full range of devices from any manufacturer.
We are constantly updating our list of hearing aids so be sure to ask about the latest developments.
Oticon Hearing Aids
Oticon hearing aids is a subsidiary of William Demant Holdings in Denmark who also owns Bernafon and Sonic Innovations. They are therefore one of the largest hearing aid manufacturers in the world and trades through retail chains including Audiolclinic and HearingLife on a national scale and Adelaide Digital Hearing Solutions at a local level.
The latest range of hearing aids released by Oticon is called the Intiga. This is a very small wireless capable hearing aid with a newly designed casing and speaker wire making this the smallest wireless hearing aid on the market
This processing unit and features are also found within Oticon's other models - Agil Pro, Agil, Acto Pro, Acto and Ino and Ino Pro range of hearing aids.
Phonak Hearing Aids
Phonak is a premium Swiss hearing aid company owned by Sonova. Phonak is one of the largest hearing aid manufacturers in the world and has always been at the leading edge of technology especially when it comes to improving a person's ability to hear speech in noise. Phonak has some very capable wireless technologies built into their hearing aids, allowing true stereo wireless transmission of sound through their Bluetooth capable devices. They also have a whole range of hearing aid compatible and free standing FM systems which extends the connectivity even further.
Their latest chipset, called the SPICE+ platform, is the words fastest hearing aid platform running at 200 million instructions per second and is based on a three processor, 6 core design. It is the only hearing aid platform capable of real time wireless broadband audio transmission and simultaneous processing.
Phonak's current top of the line hearing aids containing SPICE+ consists of the Ambra, Naida S IX, Naida S IX CRT and Smart S IX. The all have the same features but are different styles and can fit different hearing losses. With the introduction of the SPICE+ Phonak has dramatically improved their hearing aids initial acceptance and overall performance in quiet.
Phonak's range of Spice hearing aids have been extended into the basic category with the Phonak Dahlia, Smart S and Naida S I hearing aids. The offer the superior Spice+ processing, but with limited features for the less demanding client.
Starkey Hearing Aids
Starkey is an American hearing aid company that came to fame when Ronald Regan first wore their completely in the canal hearing aids. Their philosophy is to only introduce technology that has been proven through research to add client benefit. They have some novel technology such as real ear measurements through their hearing aids, they are also one of only two companies using the superior 2.4Ghz Wireless connectivity. They were also the first company to introduce the invisible in the canal style hearing aids late in 2010.
Starkey has been hard at work developing new hearing aid technology to compete with some of the best in the world. The have just released their X series during the latter part of October 2011, which includes their proprietary frequency lowering technology, called Spectral IQ. This often leads to improved speech understanding in quiet and noise for these clients.
Sonic Innovations
Sonic Innovations is an American hearing aid company. They have recently been bought by William Demant Holdings, which also owns Oticon and Bernafon and through the Sonic Innovations acquisition, obtained several hundred HearingLife clinics throughout the world.
Sonic has released their new Flip range of wireless hearing aids in Australia. This is their first wireless capable hearing aid range.
Bernafon Hearing Aids
Bernafon is a Swiss Hearing Aid Company that is part of the William Demant group who also owns Oticon and Sonic Innovations.
Bernafon hearing aids are good value to clients, they offer exceptional design and sound quality, but without the high price tag.
Widex Hearing Aids
Widex is a Danish hearing aid manufacturer that has lead the field in excellent sound quality.
Widex has released the Clear range of wireless capable hearing aids toward the end of 2010, starting with the top end Widex Clear 440 hearing aid.
Unitron Hearing aids
Unitron is originally a Canadian hearing aid company, who was bought by Sonova, the same company who owns Phonak hearing aids.
They produce good value hearing aids by packaging many of the premium technologies on the market into a lower cost, well performing hearing aid.
Siemens Hearing Aids
Siemens is a well know producer of electronic equipment in almost all industries.
Siemens has made a lot of headlines with their release of the Aquarius 701 and Aquarius 501 water proof and dustproof hearing aids in the latter part of 2011. These hearing aids promise improved reliability to those working in very humid or dusty environments and were even tested by people who swam and surfed while wearing them.
GN Resound
GN Resound has always been know for its excellent hearing aids with their natural sound quality. They are one of the most flexible Danish brands of hearing aids and their experience in wireless technology shows in their latest Alera Range of hearing aids.
GN Resound were the first hearing aid manufacturer to develop open fit hearing aids with their introduction of the Resound Air in the early 2000's. This was a remarkable breakthrough and had lead to significant improvements in hearing aid user satisfaction. Their open fit technology has now been embraced by all other manufacturers.


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