text:
Great West Life Assurance Co. Can Acquires Shares of 160,334 Smartsheet Inc (NYSE:SMAR)
Posted by MarketBeat News on Jan 22nd, 2023
Great West Life Assurance Co. Can acquired a new stake in shares of Smartsheet Inc (NYSE:SMAR – Get Rating) in the 3rd quarter, Holdings Channel.com reports. The fund acquired 160,334 shares of the company's stock, valued at approximately $5,516,000.
Several other hedge funds and other institutional investors have also made changes to their positions in SMAR. Raymond James & Associates boosted its holdings in shares of Smartsheet by 4.5% during the 1st quarter. Raymond James & Associates now owns 19,514 shares of the company's stock valued at $1,069,000 after purchasing an additional 845 shares during the last quarter. Cambridge Investment Research Advisors Inc. raised its position in shares of Smartsheet by 10.2% during the 1st quarter. Cambridge Investment Research Advisors Inc. now owns 16,949 shares of the company's stock valued at $928,000 after buying an additional 1,567 shares in the last quarter. Citigroup Inc. raised its position in shares of Smartsheet by 7.0% during the 1st quarter. Citigroup Inc. now owns 25,608 shares of the company's stock valued at $1,403,000 after buying an additional 1,677 shares in the last quarter. PNC Financial Services Group Inc. raised its position in shares of Smartsheet by 8.1% during the 1st quarter. PNC Financial Services Group Inc. now owns 10,683 shares of the company's stock valued at $585,000 after buying an additional 803 shares in the last quarter. Finally, MetLife Investment Management LLC purchased a new stake in shares of Smartsheet during the 1st quarter valued at about $431,000. Institutional investors own 89.50% of the company's stock.
Get Smartsheet alerts:
Analyst Ratings Changes
SMAR has been the topic of several recent research reports. KeyCorp initiated coverage on shares of Smartsheet in a research report on Thursday, October 13th. They set an "overweight" rating and a $46.00 target price on the stock. Barclays started coverage on shares of Smartsheet in a research report on Thursday, December 15th. They set an "equal weight" rating and a $45.00 target price on the stock. BMO Capital Markets increased their price objective on shares of Smartsheet from $38.00 to $41.00 in a research report on Friday, December 2nd. Morgan Stanley increased their price objective on shares of Smartsheet from $54.00 to $56.00 and gave the stock an "overweight" rating in a research report on Friday, December 2nd. Finally, Royal Bank of Canada increased their price objective on shares of Smartsheet from $32.00 to $36.00 in a research report on Friday, December 2nd. Four analysts have rated the stock with a hold rating and twelve have given a buy rating to the stock. According to data from MarketBeat.com, the company presently has a consensus rating of "Moderate Buy" and an average target price of $47.17.
In other Smartsheet news, insider Jolene Lau Marshall sold 6,001 shares of the firm's stock in a transaction that occurred on Wednesday, November 16th. The shares were sold at an average price of $31.45, for a total value of $188,731.45. Following the completion of the sale, the insider now directly owns 12,501 shares in the company, valued at $393,156.45. The sale was disclosed in a legal filing with the SEC, which is available through this hyperlink. In other Smartsheet news, CFO Pete Godbole sold 2,256 shares of the firm's stock in a transaction that occurred on Wednesday, December 14th. The shares were sold at an average price of $41.36, for a total value of $93,308.16. Following the completion of the sale, the chief financial officer now directly owns 18,229 shares in the company, valued at $753,951.44. The sale was disclosed in a legal filing with the SEC, which is available through this hyperlink. Also, insider Jolene Lau Marshall sold 6,001 shares of the firm's stock in a transaction that occurred on Wednesday, November 16th. The shares were sold at an average price of $31.45, for a total transaction of $188,731.45. Following the completion of the sale, the insider now owns 12,501 shares of the company's stock, valued at $393,156.45. The disclosure for this sale can be found here. Over the last ninety days, insiders have sold 15,623 shares of company stock worth $523,937. Corporate insiders own 4.58% of the company's stock.
Smartsheet Stock Performance
Shares of SMAR stock opened at $42.69 on Friday. The company's 50-day simple moving average is $36.66 and its two-hundred day simple moving average is $34.61. Smartsheet Inc has a one year low of $25.09 and a one year high of $66.74.
Smartsheet (NYSE:SMAR – Get Rating) last released its quarterly earnings data on Thursday, December 1st. The company reported ($0.30) earnings per share (EPS) for the quarter, beating analysts' consensus estimates of ($0.51) by $0.21. Smartsheet had a negative net margin of 31.75% and a negative return on equity of 45.17%. The firm had revenue of $199.58 million for the quarter, compared to the consensus estimate of $193.66 million. On average, analysts anticipate that Smartsheet Inc will post -1.66 EPS for the current fiscal year.
Smartsheet Profile
Smartsheet, Inc engages in managing and automating collaborative work. Its platform provides solutions that eliminate the obstacles to capturing information, including a familiar and intuitive spreadsheet interface as well as easily customizable forms. The company was founded by W. Eric Browne, Maria Colacurcio, John D.
Get a free copy of the StockNews.com research report on Smartsheet (SMAR)
Want to see what other hedge funds are holding SMAR? Visit HoldingsChannel.com to get the latest 13F filings and insider trades for Smartsheet Inc (NYSE:SMAR – Get Rating).
Receive News & Ratings for Smartsheet Daily - Enter your email address below to receive a concise daily summary of the latest news and analysts' ratings for Smartsheet and related companies with MarketBeat.com's FREE daily email newsletter.
iShares S&P Small-Cap 600 Value ETF (NYSEARCA:IJS) Stock Holdings Lowered by Park Avenue Securities LLC
Great West Life Assurance Co. Can Trims Holdings in Crown Holdings, Inc. (NYSE:CCK)
Daily Ratings & News for Smartsheet
Complete the form below to receive the latest headlines and analysts' recommendations for Smartsheet with our free daily email newsletter:


The goal of Catholic Youth Ministry is to help teens know and experience Christ. To do this we need to reach teens. But where are teens? Today many are on social media. Yet social media can seem daunting to those of us just a few years older. I figured I could offer some things that helped me reach teens on social media.
I don't think of myself as a social-media expert. However, somehow I've supposedly become the most influential person online in Catholic Youth Ministry – I have a Klout of 66, and Mark Hart (#2) has 64 – I should have something to say. I guess I'll try to say what I know. I hope it helps.
I only joined Twitter in November 2012 and Facebook in December of 2012 – yes, you read that right, I held out till last December to join Facebook. Somehow, I must have figured something out. I'm not 100% sure what but I try to share the little I have figured out here.
I wanted to write one big blog post but it began getting too long so I posting secrets for all types now and I'll post specific secrets for Facebook and Twitter later.
The first thing we need to realize is that social media is not some special foreign place. It's the modern town square. Don't be scared. Most of all, BE YOURSELF! If you know how to help teens in the real world, do the same in social media. Freelancers can have a special tendency to try to make themselves into someone they are not.
In social media, self-promotion is not the sin of pride. If you start a new blog, only your mom and wife will read it until somebody sees it on Twitter or Facebook. If you have a great event coming up, tell everyone.
They are following or friends so that they hear about it. Don't take this to the other extreme of being annoying and reposting your article so many times people are fed up. It's just like the real world, we like to know when our favorite team is playing but if our buddy tells us 16 times, we get annoyed.
Be ready to invest a bit of time into social media if you want to have success. Simply posting your events on Facebook does nothing if people don't read it. Posting a single event on Facebook is like posting a banner on a wall that stretches from Chicago to Detroit. Unless the people you want to read it know to stop there, it will be lost in the blur of 100 other posters. The whole value of social media is connections – active connections to be precise. Once someone connects with you, your content appears right outside their front door. However, with the exception of Twitter, this will slowly move further away unless you keep having points of contact. If Joe likes your page on Facebook, he'll likely see one or two updates but unless he comments on, shares, or likes those, Facebook will move your page to the bottom.
Cultivate your connections. If people start responding to what you're saying, show them you appreciate it. Just like we show attention when someone is talking to us in real life, we need to express our appreciation and attention to others who are talking to us on social media. If you post something, and a person responds "Amen," "Nice reflection," etc. then you should respond with a like / favorite / +1 to that comment. This is the same as nodding your head if they said this in real life. It is also nice to give likes, favorites, and +1's to other posts but its meaning is different. I like over 80% of the comments to my posts on Facebook but only about 10%-15% of the updates I read. Parishes should read what some of the youth group members are posting, and like or comment as needed. If you connect with them, they will connect with you; social media is inherently a 2-way street.
It's recommended to separate your personal and youth group account. This applies especially to parishes and less so to freelancers. You likely aren't going to be youth minister at parish X for 20 years so you want a way to separate once you're done. Plus, most of the youth group kids don't want to see your family vacation photos while your adult friends do. On the other hand, your cousin 1000 miles away doesn't need 5 reminders of your youth group events. I admit that I didn't start out this way on twitter and now I'm looking for a way to do it well.
It's very difficult to over-communicate on social media. (On twitter, it's close to impossible.) Therefore, don't just post one thing but repeat and amplify what you have to say. You shouldn't repeat the exact same thing but if you have an activity coming up and you post 4 different reminders the week before, that's completely acceptable. To get your message amplified by others, make it easy to share. If your note is complicated, few will share it but if it's simple and catchy, teens will share it among themselves (and some of them may be more credible to their peers than any of us).
You knew we had to talk about child protection norms. What can and can't we do with teens online? My basic rule: the same as real life. If you would not say something to a teen at youth group, don't say it to him online. Period. The difference here is that usually the norms require parental permission for private communication via texting or online. Responding to public posts or public tweets don't count. Usually private messaging and accepting friend requests require parental permission. If you're a parish, you have it easy, just add permission to talk to teen via social networks to the permission form. For us freelancers, I'd say that on twitter it's easy to say no DMs (5% of twitter bios say this so you won't stand out). On Facebook, do a page where everything is public, and don't accept friend requests from teens you don't know (let them follow your public posts – more on this later).
That's all for now. Stay tuned for specific secrets relating to Facebook and Twitter. To make sure you don't miss them, please go to the right hand side, (1) follow me on twitter, (2) like this page, and (3) subscribe by e-mail. As I said, it's hard to over-communicate.


EdChoice published a great compilation "The ABCs of School Choice 2018 Edition" for the all the latest news and information.
Forbes produced a great summary "Charter Schools, Vouchers, and ESAs: The Three Flavors of School Choice" although they leave out tax credit scholarships in the title.
It's always a great idea to read what your opponents are saying. NPR's "Why It's So Hard to Know Whether School Choice is Working" does just that.


EPDM Coatings - The ONLY Liquid EPDM Rubber In The World.
One of our sales representatives will be responding to you within 2 business days.
Did you know that Liquid EPDM outperforms Elastomerics and Acrylics and Urethanes 4 times longer? That means only one coat every 15 years with Liquid Rubber vs one coat every 4 years with other products!


text:
Tributes paid to globetrotting teacher of the deaf and pioneer of her field
By Charlotte Moreau Reporter
Dr Morag Clark, a pioneer of education for the deaf.
TRIBUTES have been paid to a "formidable pioneer and dedicated mentor" who dedicated her life to the field of deaf education.
Dr Morag Clark MBE, who died on April 9 aged 90, retired to Hereford in the late 1980s.
After training as a teacher in her native Scotland, Dr Clark went on to train as a Teacher of the Deaf at Manchester University.
She went on to become deputy principal and later, principal of Birkdale School for Hearing Impaired Children, where she worked until her retirement in 1986.
But Dr Clark, described by her former colleagues as having "seemingly boundless energy," brought a whole new meaning to the concept of being retired.
Driven by a wish to give deaf children the best life chances possible, she was a passionate advocate of what would become known as Natural Auralism, who believed strongly that, with appropriate aids, even the most profoundly deaf could develop spoken language in a natural way.
To that end, she went to live in Eskisehir, Turkey for two years, where she continued work with a project for deaf children at ICEM in Anadolu University.
She was also asked to lecture at the Universities of Istanbul and Ankara, and it was not long before word of the success of her work began to spread and Dr Clark was being asked to carry out similar work in many other countries.
For more than 20 years she travelled almost constantly in her voluntary role as a consultant, working in at least 14 different countries.
So formidable was her itinerary that there was one year in which she only spent a total of six weeks in her Hereford flat!
Despite this, she still made time to carry out acts of kindness such as taking a group of homesick African students to her flat for a good meal and to watch football on her television.
Dr Clark was justifiably honoured for her work in several ways.
Awarded an MBE for international services to the hearing impaired in 1989, she was also the recipient of three awards from the Alexander Graham Bell Association and awarded an Honourary Doctorate by Anadolu University.
For Dr Clark, however, the greatest reward was always the pleasure of chatting with young deaf people whose families she had helped and who were leading fulfilling lives making their way independently in the world.
"Those of us who had the pleasure of knowing and working with Morag are still awed by all that she achieved, but above all, we will remember what a wonderful friend and colleague she was," said former colleague, Liz Rothwell.
"The field of education of the deaf will be immeasurably poorer without Morag Clark, as were the lives of those of us who were privileged to have her friendship."

redpajama_set_name: RedPajamaCommonCrawl


text:
Home/News/61-year-old Asian man in critical condition after brutal assault caught on video
61-year-old Asian man in critical condition after brutal assault caught on video
Marlene Lenthang, ABC NewsApril 24, 2021
New York Police Department
(NEW YORK) — A 61-year-old Asian man was left in critical condition after he was brutally thrown to the ground and kicked in the head in New York City in a harrowing attack captured on surveillance video.
The incident unfolded Friday night at 8:20 p.m. on the corner of Third Avenue and E.125th Street in Manhattan.
The victim was approached from behind by an unknown individual who "struck the victim in the back causing him to fall on the ground," the New York Police Department said in a statement.
The individual then "kicked the victim multiple times in the head" and fled on foot, police said.
The attack was caught on surveillance video footage and shows a man repeatedly kicking the victim on the ground.
Paramedics responded and the victim was transported to NYC Health & Hospitals/ Harlem, where he is listed in critical but stable condition.
The NYPD's Hate Crime Task Force is investigating the incident.
Police are asking for the public's help in identifying the man behind the attack and shared photos of him from the surveillance footage.
Police described the individual as an adult male with a dark complexion, last seen wearing a black jacket, black pants, white sneakers and a multi-colored baseball cap.
Anyone with information in regard to this incident is asked to call the NYPD's Crime Stoppers Hotline at 1-800-577-TIPS (8477) or for Spanish, 1-888-57-PISTA (74782). The public can also submit their tips by logging onto the Crime Stoppers website at WWW.NYPDCRIMESTOPPERS.COM, or on Twitter @NYPDTips.
Marlene Lenthang, ABC News
Snake River among top 10 most endangered rivers in the US, conservation group says
Former NFL player allegedly left cellphone at scene of mass shooting: Warrants
American Jewish groups, leaders send support to a Texas synagogue after hostage situation–and look to what comes next


Save the Date! Linda Ott will be Speaking on Tuesday, December 5th.
Please be reminded that Linda Ott from Sustaining Way will be speaking at Temple B'nai Israel on Tuesday, December 5 at 12:00 noon on the topic "Tis a gift to be simple: How to Experience the Winter Celebrations in an Eco-Conscious Manner". Please see attachments. We hope you will join us for this informative lecture. Lunch will be served as well. RSVP to the temple at 864-582-2001.
Linda comes to the SCIPL (South Carolina Interfaith Power & Light) Director role most recently as a graduate of Columbia Theological Seminary with a Masters of Theology in Creation Care. Linda's vocation life journey included the military, social work, business, and law. Her passion for creation care stems from her time at Drew Theological School, where she completed a Masters of Divinity and discovered her passion for understanding the intersection of faith and the care of creation.
As a people of history it is essential to recall how it is we got from there to here; at times in celebration, at times in consolation. The review by one of my long time colleagues and classmates reviews the partition plan and the little the world hesitatingly gave our people after the Shoah, the Holocaust.
Seventy years ago today, November 29, 1947, the newly formed United Nations voted to partition Palestine into two states, one Jewish state and one Arab state. The Jewish people led by David Ben Gurion accepted the Partition Plan but it was rejected by all Arab States.
If Jewish destiny is placed in the hands of Jews for which Judaism is only a result of persecution, chance or a random gesture of kindness, it will surely sink into ignorance and indifference… if the steering wheel is left in the hands of Jews whose Judaism is an inner necessity, a covenant in their soul, who wish to continue to path of Jewish glory – both people and culture – only then can we be sure that the necessary institutions to enrich our lives, most importantly the synagogue and school – and particularly the school – will be established.
This link includes a speech given by Rabbi Silver before the United Nations on November 29, 19487 beginning at two minutes and thirty seconds to nine minutes and 40 seconds. You can hear, as well, the roll call vote of the nations voting on the Partition plan beginning at 11 minutes and 50 seconds.
The final vote was 33 to 13 with 10 abstentions in favor. The map as determined by the partition plan can be seen here and there is also a link to what countries voted for, against and abstained.
7 Israelis Who Have Made a Deep Impact on Life in the U.S.
When Natalie Portman was named the 2018 winner of the $1 million Genesis Prize, known as the "Jewish Nobel," it was in part an acknowledgement of her Israeli roots.
While the Oscar-winning actress mostly grew up in the United States, Portman — née Herschlag — is also Israeli. Her father, Avner Herschlag, grew up in Israel, and her mother married him there. The family moved to the U.S. when she was 3.
Human beings are not robots — sometimes they make irrational decisions, and they are always complex.
That idea might sound like common sense, but before economist and psychologist Daniel Kahneman started his Nobel Prize-winning work in the 1970s, economics plugged people into equations just as they would other sets of numbers. Kahneman's work with his Israeli research partner Amos Tversky (he died from cancer in 1996) on concepts such as cognitive biases and prospect theory helped change that and effectively launched the field of behavioral economics — which in turn has influenced several other fields. His 2011 book Thinking, Fast and Slow, which summarized much of his research, has sold more than a million copies around the world.
Gadot, a former Miss Israel who had previously had small roles in a few Fast and Furious movies, pulled it off, turning Wonder Woman into a blockbuster while inspiring woman around the world. Off screen, she has earned praise for refusing to work with a producer accused of sexual misconduct.
Gadot, now 32, grew up near Tel Aviv, served in the Israeli army and only recently moved to Los Angeles. Guest-hosting Saturday Night Live, she spoke briefly in Hebrew to her family back in Israel — no small gesture in an era of anti-Israel boycotts.
How did a guy who wrote music for cartoons become one of the most powerful media moguls, political donors and pro-Israel forces in the U.S.? The answer begins with Power Rangers.
Saban, who was born in Egypt but grew up in Tel Aviv, joined a band after serving in the Israel Defense Forces. He then became a concert promoter and eventually immigrated to the U.S. in 1983, where he lived a comfortable life as a cartoon music composer. He discovered the Power Rangers, which was first a show in Japan, in a hotel room during a business trip and instantly fell in love with the campy teenage superheroes.
He immediately bought the rights to the show, but shopped the idea around to American executives for years before one, Margaret Loesch at Fox, bought it. Since the show's debut in 1993, "Mighty Morphin Power Rangers," has netted billions of dollars in TV profits and merchandise. Saban grew his fortune by investing in other media ventures and became one of Los Angeles' most generous philanthropists (among other things, a research clinic at Children's Hospital Los Angeles bears his name).
Saban, 73, has also poured millions into Democratic politics over the years — notably he gave Hillary Clinton's failed presidential campaign $15 million. As a staunch Israel backer, he supports and hosts the annual Saban Forum on Israel, which brings together political leaders from the U.S. and Israel to debate topics related to the Jewish state.
Saban also was a key supporter of what became the Israeli-American Council, which aims to boost the Israeli and Jewish identity — and political clout — of Israelis living here.
Perlman's introduction to the international spotlight came in 1958, when the prodigy performed on The Ed Sullivan Show at the age of 13. But he had already been playing the violin for about a decade — and had been living with paralyzed legs from a bout of polio for nearly the same amount of time.
Fast forward several decades, and the New York-via-Tel Aviv native has earned 16 Grammy Awards, a Presidential Medal of Freedom and the title of "world's greatest living violinist." As a performer, Perlman is not only known for his smooth tone and effortless finger speed — his energy and love of playing, usually communicated through a signature wide grin, has helped make him an international music icon. He also fosters young musicians through the Perlman Music Program, which he founded with his wife in 1994.
Outside of music, Perlman has passionately advocated for rights for people with disabilities. After being awarded the Genesis Prize in 2015, he pledged his winnings toward improving the integration of people with disabilities into Israeli and American society — with a specific focus on his adopted hometown of New York.
Gabe Friedman is the editorial assistant at JTA.
Wofford College Lighting of the Hanukkah Menorah, November 28th!
Tonight is the Winter Lighting program in Leonard Auditorium which starts at 6:30. After the indoor festivities, the audience will move outside to watch the lighting of Wofford's 6-foot menorah in front of Old Main. That menorah lighting typically occurs between 7:20-7:30 depending on when the indoor program ends. The menorah lighting is a quick ceremony to mark the beginning of the holiday season. Please join the Rabbi tonight at Wofford College for the lighting.
I love Thanksgiving. Maybe because I grew up in Canada, and as a child, never had the experience of participating in this uniquely American holiday. But this year was the 31st Thanksgiving I spent with my husband's family (since my family doesn't celebrate Thanksgiving, it is the one holiday we never had to share between our two families) and the joy and sense of awe I feel as we all gather around the table has not diminished in all those years.
And yet, it always feels a little odd that there is no ritual to begin the meal. On all of the other holidays where we gather around the festive table – Rosh HaShanah or Passover or even a special Friday night Shabbat dinner – we begin with Kiddush, blessing the wine, and marking the specialness, even holiness, of the day.
So when the Belfer Center for Innovation & Social Impact at the 92nd Street Y in New York inaugurated #GivingTuesday in 2012, I finally found my way of infusing Thanksgiving with some holiness. For what could be more sacred than ensuring that tzedakah, giving to further the work of world-repair, be tied to the holiday?
And when the members of the household eat and drink, they are obligated to feed strangers, orphans and widows as well as all other poor people. However, if they lock their doors and eat and drink with their family and do not feed the poor and others going through hard times, this is not the joy which was commanded, but merely satisfying their stomachs.
We cannot truly celebrate a holiday, according to Maimonides, if we limit our celebration and feasting to ourselves alone. Rather, we must tend to the needs of others in our community.
But I believe our tradition has even more to teach us about giving.
The Hebrew word tzedakah, which is usually translated as charity, philanthropy, or giving, has a much more profound meaning. It comes from the same root as the word tzedek, justice, and implies that giving charity alone is not nearly enough. Jewish tradition demands that, particularly at a moment when we are enjoying our own material blessings, we must do all we can to ensure that our society is built on the pillars of economic and social justice.
Tzedakah cannot be translated because it joins together two concepts that in other languages are opposites, namely charity and justice. It is the idea that no one should be without the basic requirements of existence, and that those who have more than they need must share some of that surplus with those who have less. This is fundamental to the kind of society the Israelites were charged with creating, namely one in which everyone has a basic right to a dignified life and equal worth as citizens in the community.
In the midst of the post-Thanksgiving glow, and the pre-holiday season excitement, we should use #GivingTuesday as an opportunity to live out one of the most fundamental teachings of our tradition. Whether we choose to support a local food pantry or an international humanitarian relief agency, civil rights organizations in North America, or those fighting for religious pluralism in Israel, we must all do our part to strengthen social and economic justice in the world.
#GivingTuesday may be a new idea, but let us seize this moment, and commit ourselves to an ancient value, true tzedakah.
Support the Union for Reform Judaism, which upholds the principle of tikkun olam (repairing the world) by strengthening communities across North America and creating a world in which all Jewish people can participate and be proud.
Sisterhood General Meeting! Please join us Wednesday, November 29 at 7:00 for wine, nibbles and socializing. Come to hear how we did with the bake sale. We will also elect the new board.
The horizon of this world binds my perceptions.
What I see and what I know are tied to my awareness.
That I accept with humility and thanksgiving.
To become an instrument of Divine bounty.
With reverence for Your creation.
Providing bounty to be shared.
© 2010 Alden Solovy and tobendlight.com. All rights reserved.


Model MITMV6835 extends the capabilities of our MIT6830 vacuum brake bleeder by including our MITMVA6832 Clamp-on Auto-refill Kit. While the bleeder can evacuate up to 2 quarts (2 liters) per minute, the refill kit ensures the master cylinder does not run dry of new fluid, making this kit an extremely efficient method of bleeding hydraulic brake and clutch systems. Features Swivel air inlet with quick disconnect. Two refill kit styles ensure a fit to all makes and models of vehicles.
Many people regard the Mityvac MV6835 as one of the best brake bleeder kits on the market. One of the most distinguishing factors of this product is its large reservoir that can hold an impressive oil capacity of 1.8 liters.
With such a big reservoir capacity, it will have no problem working with an oversized vehicle like an RV. Additionally, the hose is long at 6 feet so as to provide more flexibility when bleeding the brake system in small vehicles that usually have a tight space to work.
Moreover, this brake bleeder provides sufficient pressure so as to bleed the braking system quickly and easily. In addition, you get many accessories like a hanging hook for hanging the hose in mid-air for better ease of use.
Of course, this bleeder also comes with all vital accessories like a hose, tight fit adapters as well as a 40 oz. refill cylinder.


text:
NASDAQ a Hair Short of a New High, Dow Maintains Streak
Jul 19, 2018 | US Stocks Review
The gains may not seem spectacular but with all of the cross and headwinds, the market isn't doing half bad! All the NASDAQ needed to do was climb a smidgen, and it would have found itself at a new all-time high but it just didn't have the stamina. That notwithstanding, with trade fears rampant, mid-terms nearing, and Trump regaining his footing after his botched Russian summit, there's not much that we can complain out. The Dow is experiencing its longest winning streak in a month. That notwithstanding, the market has been unforgiving, and expects perfection from reporting companies. Any miss, revenue or earnings, or a miss in future projections, can send a stock sliding. The stakes are high, so before you assume a stock will rally, recall that "God is in the details," and if top and bottom line numbers are right on the money, but projections disappoint, a stock could go through the shredder, and that's just what we're seeing!
One of the big focal points yesterday was the release of the anecdotal account of the Fed's 12 member districts, otherwise known as the Fed Beige Book. The picture was optimistic, offering market support. One of the highlights was a swiftly growing domestic economy, though one with a ceiling from above given a paucity of skilled worker. Add to that, noted the Fed, the rising cost of raw materials, and an ongoing expansion could be called into question. All-in-all, of the 12 districts, 11 regions grew at a pace that was "modest" or faster, with only those states in the St. Louis region seeing "slight" growth.
On the political front, Capitol Hill has been awash with tension as lawmakers themselves, from both sides of the table, try to battle Trump's go it alone approach. Though Trump has boasted that "cars" – plain and simple – are his path to victory, American and foreign manufacturers alike have repudiated that approach, going for the jugular, teaming up to fight Trump's one-upmanship negotiating tactics. The Senate already voted 88-11 to give Congress more power to reject Trump's contentions that tariffs are needed on national security grounds. And yesterday, the House sent the Oval Office a joint letter stating the following: "We do not believe the imports of automobiles and automotive parts pose a national-security threat," adding, "Rather, we believe the imposition of trade restrictions on these products could undermine our economic security." Likewise, the auto industry has sent a letter of its own: "Raising tariffs on auto and auto parts would be a massive tax on consumers, who buy or service their vehicles."
We have seen a solid trend in so far as earnings reports go. Altogether, 90% of reporting companies have seen bottom line beats, earnings outstripping analyst forecasts. It's hard to say, though, that we've seen large-scale momentum carrying the market higher. Over and beyond earnings, Fed Chair, Jerome Powell fleshed out again the comments he had made the day before, more pointedly, that we can plan on seeing moderate, gradual rate hikes. In Wednesday's testimony, the Fed Chair stated that there are few signs of an impending economic recession. For now, he noted, the Fed will continue to hike rates every 3 months.
Another development that may or may not drive markets from hereon in is the statement made yesterday by Trump's advisor, Larry Kudlow, namely, that further pro-growth measures could be around the corner. We'll have to see if that's translated into any policy changes. At this point it's key to note that Trump, perhaps less skilled at creating global coalitions or consensus in his own party, is nonetheless a master of branding himself and forging expectations that fuel market growth. When the market started falling, he started tweeting, something unprecedented for presidents in recent years or decades. With the market climbing significantly since Trump's ascension, it might be premature to expect a bear market, especially when all of the indicators and the Fed itself saying otherwise.
One landmark decision that's worth noting was the turnaround made by the board of Berkshire Hathaway. The company has been sitting on an enormous pile of cash, cash that it has done nothing with. Buffett has failed in his "elephant hunt," those moneys doing little to return value to investors. Yesterday, though, the company board approved a share buyback beneath the threshold of 1.2x book value, a monumental shift in the thinking at one of the most storied and celebrated companies of all time. Berkshire soared 5% on the day, the company now allowed to buy back shares at a higher denominator. The gap between book value and intrinsic value growing for years, the company has failed to provide returns adequate to many, and therefore, failed deals and all, it seems like Buffett is searching for elephants in his backyard, or home market.
Buffett, in last quarter's financial letter, wrote as follows: "Some people have come close to calling [buybacks] un-American — characterizing them as corporate misdeeds that divert funds needed for productive endeavors. That simply isn't the case: Both American corporations and private investors are today awash in funds looking to be sensibly deployed."
Prudential Financial chief market strategist, Quincy Krosby, noted, "Earnings reports so far have been solid and despite today's disappointing housing starts, economic data have also been trending higher, providing support for markets."
Market Summary: The S&P 500 was up 0.2%. The catalyst for the gains were the financial sector which tacked on 1.5%, and industrials, which saw gains of 1.1%. The blue chip Dow climbed 0.3%. making it the index's 5th consecutive winning session. It was the index's longest winning streak since an 8-day streak came to an end on May 14th.
Economic Calendar: Today won't lack for economic events, with weekly jobless claims and the Philly Fed report coming out at 8:30. Then at 10:00, the Leading economic indicators report will be released.
Thursday's Hot Stocks: CSCO, IBM, AA, EBAY, AXP
IPOs: AFGL, ALLK, CNST
Index Last
DJX 25,199 +79.40 +0.32%
SPX 2,816 +6.07 +0.22%
NASDAQ 7,854 -0.67 -0.0086%


text:
Q: Calculate Process Time Excel I am looking for some assistance in calculating the time taken between 2 dates.
I have in column "A" item name and each item can have up to 5 completion stages called item status in column "B". Each completion stages has a time/date stamp in column "C".
Question: How do I calculate the time between each stage?
I have tried with =Networkdays but I'm not sure how to add the criterias to the formula?
Criterias: Same item name and the calculation need to be in the right seqence (time between Step 1 and 2, time between step 2 and step 3 etc.)
This is the formula I've tried to use but I am not sure how to add the criterias
=NETWORKDAYS(C2;C3)

A: Use when sorted:
=IF(A2=A3,NETWORKDAYS(C2,C3),"-")

If not sorted.
=IF(MINIFS(C:C,A:A,A2,C:C,">"&C2)>0,NETWORKDAYS(C2,MINIFS(C:C,A:A,A2,C:C,">"&C2)),"-")

redpajama_set_name: RedPajamaStackExchange


Waterville, famous as an angling centre, also has much to offer the general tourist. There is a fine sandy beach on the shore of Ballin-skelligs Bay, and a championship golf course (18). For years, Waterville was famous for being Charlie Chaplin's favorite retreat.


Well I bought the codex a couple of days ago and over all I'm very happy with it it's 96 page over 65 page for the old one, the typical scenario has followed with the codex as with recent codexs....troops have got cheaper....characters have got more expensive.
Hormagaunts - have dropped to 6pts each almost half, but have lost alot and I mean alot of their speed. They fleet using 3D6 picking the highest, but are no longer beasts and hence assault the usual 6", they've also dropped to WS3 but increased to I5, they retain a single set of scything talons allowing them to re-roll misses in CC on any roll of 1. They can be given toxin sacs for +2pts meaning they always wound on 4+ and/or adrenal glands for +2pts giving them furious charge. I'll miss my 30" charge...not convinced at a 18" charge.
With these stats I think I'd prefer to save the pts, plus S4 weapons can damage vehicles.
Gargoyles - There 6 pts, 1 pts more than a termagants have exactly the same stats but are jump infantry and also have "blinding venom" rolls to hit on 6 in CC autowound. Bet there cheap as GW want to sell there lovely new plastic minitures.
Genestealers - Have dropped 2pts but can no longer be given 4+ armour.
Ripper swarms - Same points but crappier.
Carnifex - 160pts SW3 S9 now and can't be given 2+ armour, same Carnifex priced up previously 101pts.
Hive Tyrant - 170pts WS8 BS3, would be priced at 114pts admittably only WS6 but otherwise the same.
Warriors - 30pts basic with Devourer and Scything talons have 3 wounds and 4+ armour S4 I4 A3 (Not immune to instant death), same points as old warriors.
Lictor - 65pts dropped 15pts have 3 wounds, deploy like Marbo (Imp guard) which means he can't charge upon first appearing, also now only +1 to cover not +2.
Anyway that's my review on some of the main models within the codex, overall I'm very happy with the new codex, just the price increase on the Hive Tyrant and Carnifex is a little much IMO.
So your swarm can now be a bit bigger but takes longer to scuttle across the board then?
Pretty much just the Hormagaunts are the slower ones, you see my old tactic was to race the Hormagaunts across the table, they typically run out of Synapse range, but by being in CC they block lines of sight and tie up front line troops in CC. It typically meant you only had 1 round of shouting to stop the Hormagaunts, now I reckon you'll have between 1 and 2. I'm sure Al will like the new slower Hormagaunts.
Actually a little supprised to hear you say that Al thought you'd be rejoicing.


text:
VIRUS TRACKER - Dec. 2: 78 New COVID-19 Cases In Hawaii - Honolulu Civil Beat
Nonprofit News Powered By Margery Bronster Victor Craft Virginia Loo Jesse Vega-Frey Andrea Wagner Readers Like You
Alicia Lou/Civil Beat/2022
Plastic Water Bottles May Be Next Ban In Hawaii's War Against Pollution
VIRUS TRACKER — Dec. 2: 78 New COVID-19 Cases In Hawaii
Many of the cases in recent weeks have been associated with two correctional facilities.
By The Civil Beat Staff / December 2, 2020
Health officials reported 78 new COVID-19 cases statewide on Wednesday, including 63 on Oahu, one on the Big Island, one on Kauai, four on Maui. Nine residents were diagnosed out of state.
No new deaths were reported Wednesday. The state's official death toll is 244. Civil Beat calculates at least 259 people have died from the disease in Hawaii, including 15 deaths on Hawaii island that are pending medical verification by the state. The Hawaii County Civil Defense has recorded 49 deaths on the Big Island.
The cases announced Wednesday reflect laboratory tests electronically reported to the Department of Health through Sunday at 11:59 p.m.
Health officials have documented 1,250 new cases during the last two weeks. Many cases have been associated with ongoing outbreaks at correctional facilities.
Other recent clusters of infection have affected coffee shop employees, restaurants coworkers, and a bar patron and bartender. On Oahu, DOH investigators are tracking four clusters in unspecified educational settings that have infected at least two dozen people within the span of two weeks.
Reporting Lag
Civil Beat's virus tracker is updated when the state releases its daily numbers. The state posts new data daily at noon. The daily reports reflect laboratory results collected up until 11:59 p.m. two days prior, which means there is a 36-hour lag between when results are submitted to the Department of Health and when the state reports them publicly.
According to Janice Okubo, spokeswoman for the Hawaii Department of Health, the COVID-19 case reports are based on test results from laboratories statewide and include test results from both residents and tourists tested within the state.
The results of Hawaii residents who test positive out of state also are included, as well as cases confirmed among military service members and their family, although military cases are not specifically indicated.
If a visitor takes a test outside of Hawaii, those test results are reported to their home state health authorities and would not be reflected in the Hawaii health department's reports. In some cases, rapid point-of-care testing results are not submitted to the DOH and are not included in these daily reports either.
For more information, check this Hawaii Department of Health COVID-19 site or this state site, and the Hawaii Data Collaborative COVID-19 Tracking site. The Hawaii Emergency Management Agency also provides this comprehensive dashboard.
Source: Hawaii Department of Health, Hawaii Emergency Management Agency


text:
Norway competed at the 1998 Winter Olympics in Nagano, Japan.

Medalists

Alpine skiing

Men

Men's combined

Women

Women's combined

Biathlon

Men

Men's 4 × 7.5 km relay

Women

Women's 4 × 7.5 km relay

 1 A penalty loop of 150 metres had to be skied per missed target.
 2 One minute added per missed target.

Bobsleigh

Cross-country skiing

Men

 1 Starting delay based on 10 km results.
 C = Classical style, F = Freestyle

Men's 4 × 10 km relay

Women

 2 Starting delay based on 5 km results.
 C = Classical style, F = Freestyle

Women's 4 × 5 km relay

Curling

Men's tournament

Group stage
Top four teams advanced to semi-finals.

|}

Medal round
Semi-final

Bronze medal game

Women's tournament

Group stage
Top four teams advanced to semi-finals.

|}

Contestants

Freestyle skiing

Women

Nordic combined

Men's individual

Events:
 normal hill ski jumping
 15 km cross-country skiing

Men's Team

Four participants per team.

Events:
 normal hill ski jumping
 5 km cross-country skiing

Ski jumping

Men's team large hill

 1 Four teams members performed two jumps each.

Snowboarding

Men's halfpipe

Women's halfpipe

Speed skating

Men

Women

References
 Official Olympic Reports
 International Olympic Committee results database
 Olympic Winter Games 1998, full results by sports-reference.com

Nations at the 1998 Winter Olympics
1998
1998 in Norwegian sport

redpajama_set_name: RedPajamaWikipedia


to view the complete exam announcement.
The state holds a civil service exam every two years to become an officer. The next exam takes place in 2019.
In order to be appointed to the State Police Academy, you must, complete the Civil Service Exam, Successfully pass the Physical Fitness Test, Successfully pass the Medical Examination, Successfully pass the Psychological Examination, Successfully pass a background investigation.


text:
Michael Edward "Mike" Goddard  is a professorial fellow in animal genetics at the University of Melbourne, Australia.

Education
Goddard was educated at the University of Melbourne where he was awarded Bachelor of Veterinary Science (BVSc) and PhD degrees.

Awards and honours
Goddard was elected a Fellow of the Australian Academy of Science (FAA) in 2011.

He was elected a Fellow of the Royal Society (FRS) in 2015. His certificate of election reads:

In 2018 he won the  John J. Carty Award for the Advancement of Science.

Selected publications

References

Living people
Fellows of the Royal Society
University of Melbourne alumni
Academic staff of the University of Melbourne
British geneticists
Australian geneticists
Fellows of the Australian Academy of Science
Year of birth missing (living people)


I have no willpower. None whatsoever. And when it comes to a case full of chocolates, all shiny and beautiful, I usually err on the side of lavish and try one of everything. Now, if it's a chocolatemaker in Vancouver, I can pace myself and try a few on each visit. But when travelling, I usually get one piece of everything that's available. It's a hard-knocked life.
So when I was in Toronto, I bought a giant box of truffles from Soma Chocolatemaker. They range from classic (fleur de sel caramel, orange marzipan) to modern (single-origin flavours) to unusual (Douglas Fir, olive oil). I'm impressed at the range of flavours, but more importantly, with the execution. Each piece was perfect, each shell was uniform and thin, and the fillings all delivered what they promised.
The standout, though, was Sparky: gianduja laced with poprocks. I know it sounds gimicky. But the gianduja (a combination of caramelized hazelnuts and milk chocolate) was the perfect vehicle to deliver those long-lost childhood exploding candies. It was entirely delightful, entertaining, and whimsical – which is, actually, what the experience of tasting chocolate should be like. It's a seasonal truffle, but maybe if we all request it en masse, it'll win a permanent spot on the chocolate menu.
That sounds really cool. I secretly like poprocks! Good thing you got some of everything or you may have missed out on this one!
Sometimes just a little piece of chocolate isn't enough. Sometimes you just need the whole pound of chocolate goodness.
justalittlepiece – Heh. You read the part about me not having willpower, right?
Kelly – I'm totally making those lollipops. And I could probably help you get rid of those pastry rocks, one way or another. How big are we talking? Maybe you could cast bars. I could be convinced to split an order with you at Christmas.


text:
JB Special Collection
JB Classic Spotlights
PHYSIOLOGY AND METABOLISM
Acetobacter aceti Possesses a Proton Motive Force-Dependent Efflux System for Acetic Acid
Kazunobu Matsushita, Taketo Inoue, Osao Adachi, Hirohide Toyama
Kazunobu Matsushita
Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Yamaguchi 753-8515, Japan
For correspondence: kazunobu@yamaguchi-u.ac.jp
Taketo Inoue
Osao Adachi
Hirohide Toyama
DOI: 10.1128/JB.187.13.4346-4352.2005
Acetic acid bacteria are obligate aerobes able to oxidize ethanol, sugar alcohols, and sugars into their corresponding acids. Among them, Acetobacter and Gluconacetobacter species have very high ethanol oxidation capacity, leading to accumulation of vast amounts of acetic acid outside the cell. Since these bacteria are able to grow in media with high concentrations of acetic acid, they must possess a specific mechanism such as an efflux pump by which they can resist the toxic effects of acetic acid. In this study, the efflux pump of Acetobacter aceti IFO 3283 was examined using intact cells and membrane vesicles. The accumulation of acetic acid/acetate in intact cells was increased by the addition of a proton uncoupler and/or cyanide, suggesting the presence of an energy-dependent efflux system. To confirm this, right-side-out and inside-out membrane vesicles were prepared from A. aceti IFO 3283, and the accumulation of acetic acid/acetate in the vesicles was examined. Upon the addition of a respiratory substrate, the accumulation of acetic acid/acetate in the right-side-out vesicles was largely decreased, while its accumulation was very much increased in the inside-out vesicles. These respiration-dependent phenomena observed in both types of membrane vesicles were all sensitive to a proton uncoupler. Acetic acid/acetate uptake in the inside-out membrane vesicles was dependent not on ATP but on the proton motive force. Furthermore, uptake was shown to be rather specific for acetic acid and to be pH dependent, because higher uptake was observed at lower pH. Thus, A. aceti IFO 3283 possesses a proton motive force-dependent efflux pump for acetic acid.
Acetic acid bacteria are obligate aerobes that belong to the α-Proteobacteria and have a strong ability to oxidize ethanol, sugar alcohols, and sugars into their corresponding organic acids. Such oxidation reactions are traditionally called oxidative fermentation, since they involve incomplete oxidation of these compounds. These bacteria accumulate the corresponding incomplete oxidation products in large quantities in their surrounding environment. Acetic acid fermentation is a typical oxidative fermentation and is used industrially to produce vinegar.
Acetic acid bacteria were divided into five to six genera (16, 21, 22), of which Acetobacter and Gluconacetobacter species can tolerate high concentrations of acetic acid, which explains their use in vinegar production. However, in addition to their ability to oxidize ethanol, Acetobacter and Gluconacetobacter species can further oxidize acetic acid, a phenomenon called acetate overoxidation. In ethanol culture, these bacteria exhibit three growth phases; they first grow by completely oxidizing ethanol into acetic acid, then stop growing for a period before they resume growing by utilizing the accumulated acetic acid during the overoxidation phase (1, 13, 14). This overoxidation had led to confusion in understanding the mechanism responsible for acetic acid resistance, because it may also cause acetic acid resistance by consuming acetic acid.
For Acetobacter aceti, the isolation of acetic acid-sensitive mutants (2, 3) and proteome analysis of acetic acid-induced proteins (10) have been performed without regard for this overoxidation-dependent phenomenon. The overoxidation of acetic acid is caused by increased activity of tricarboxylic acid cycle enzymes and acetyl-coenzyme A synthetase (14) and also by increased NADH oxidase activity in the respiratory chain (unpublished data). The acetate resistance genes isolated from A. aceti mutants encode citrate synthase and a protein involved in acetate assimilation (2, 3), and aconitase is increased in the presence of acetic acid in the culture (10). The genes and proteins are involved as part of the acetate assimilating system active during the overoxidation phase. However, since there is no acetate utilization during ethanol oxidation and the diauxic phase of Acetobacter spp. (1, 13, 14), another mechanism must be responsible for acetic acid resistance. The most plausible mechanism is an efflux pump in the cytoplasmic membrane that pumps acetic acid from the cytoplasm to the outside. However, no candidate efflux protein has been detected in acetic acid bacteria, although as many as 50 protein species have been observed by proteomic analysis to increase after acetic acid adaptation in A. aceti (5, 17).
To examine whether an efflux pump is responsible for acetic acid resistance in acetic acid bacteria, we measured acetic acid and acetate transport in intact cells of A. aceti IFO 3283. In addition, we prepared right-side-out (RSO) and inside-out (ISO) membrane vesicles and measured acetic acid or acetate accumulation or uptake. The results suggested that A. aceti IFO 3283 has an efflux pump specific for acetic acid which is driven by a proton motive force but not by ATP.
Materials.[1-14C] sodium acetate (9.25 MBq, 50 mCi/mmol, 200 μCi/ml) was purchased from Amersham, and 3,3′-diisopropylthiodicarbocyanine [diS-C3-(5)] and bis(1,3-diethylbarbituric acid) pentamethin oxonol [diBA-C2(5)] were purchased from Molecular Probes Inc. 9-Aminoacridine (9-AA) was from Sigma. All other chemicals were commercial products of guaranteed grade.
Bacterial strains and growth conditions. A. aceti IFO 3283 was used in this study. Cells were maintained on an agar slant which was prepared by adding 1.7% agar and 0.5% CaCO3 to a potato medium (6) consisting of 5 g of glucose, 20 g of glycerol, 10 g of yeast extract, 10 g of polypeptone, and 100 ml of potato extract in 1 liter of tap water. Cells from the slant were inoculated in 5 ml of the potato medium and cultivated at 30°C with rotary shaking at 200 rpm until a Klett unit (Klett Summerson photoelectric colorimeter, Klett MFG Co.) of 250 was reached. The seed culture was then transferred to 100 ml of glycerol medium consisting of 0.2% each of glycerol, yeast extract, and polypeptone in a 500-ml Erlenmeyer-flask and then grown at 30°C with rotary shaking at 200 rpm to the late exponential phase.
Preparation of membrane vesicles.Cells were collected by centrifugation at 7,000 × g for 10 min and washed twice with 50 mM potassium phosphate buffer (KPB, pH 6.5) including 5 mM MgCl2. Spheroplasts were prepared by essentially the same procedure used for Acetobacter lovaniensis (originally called A. aceti) IFO 3284 (7). The washed cells were resuspended in 0.1 M Tris-HCl (pH 8.0) containing 10% sucrose and 1% sodium EDTA (pH 10.6), and then lysozyme and DNase were added to final concentrations of 1 mg/ml and 6 μg/ml, respectively. The suspension was incubated for 1 h at 30°C with shaking and then centrifuged at 7,000 × g for 10 min to isolate spheroplasts.
To prepare RSO membrane vesicles, the precipitated spheroplasts were suspended with a minimal volume (0.7 g/ml) of 50 mM KPB (pH 6.5) containing 10% sucrose and rapidly diluted into 200 ml of 50 mM KPB (pH 6.5) containing 5 mM MgSO4 and 6 μg/ml of DNase to lyse by osmotic shock. After incubation for 30 min at room temperature, the solution was centrifuged twice at 7,000 × g for 10 min, and the supernatant was centrifuged at 48,000 × g for 30 min. The precipitate was suspended in 50 mM KPB (pH 6.5) containing 5 mM MgSO4 at 0.5 g wet weight/ml and stored at −80°C as RSO membrane vesicles.
To prepare ISO membrane vesicles, spheroplasts were suspended in 50 mM KPB (pH 6.5) containing 5 mM MgSO4 and 6 μg/ml of DNase at a concentration of 0.7 g wet spheroplasts/ml. The suspension was passed through a French pressure cell at 5,000 lb/in2, and after removing cell debris by centrifugation at 9,000 × g for 10 min, the supernatant was centrifuged at 105,000 × g for 90 min. The precipitate was suspended in 50 mM KPB (pH 6.5) containing 5 mM MgSO4 at 0.2 g wet weight/ml and stored at −80°C as ISO membrane vesicles.
Measurement of the membrane potential (Δψ) and pH gradient (ΔpH).Generation of Δψ with RSO and ISO membrane vesicles was monitored by following fluorescence quenching of diS-C3-(5) for inside negative potential and diBA-C2(5) for inside positive potential, respectively (8). Measurement of Δψ (inside negative) was performed at 25°C in a reaction mixture (total, 1 ml) consisting of 50 mM KPB (pH 6.5), 5 mM MgSO4, 1 μM diS-C3-(5), and RSO membrane vesicles (≈100 μg of protein), and fluorescence emission was monitored at 670 nm with excitation at 622 nm. For the measurement of Δψ (inside positive), the reaction mixture (1 ml, total volume) contained 50 mM KPB (pH 6.5), 5 mM MgSO4, 2 μM diBA-C2-(5), and ISO membrane vesicles (≈500 μg of protein). The reaction was carried out at 25°C with fluorescence emission at 614 nm and excitation at 588 nm. In either case, the reaction was started by the addition of an electron donor, such as lactate, followed by the addition of 0.1 μM nigericin, and stopped by the addition of 1 μM valinomycin. The Δψ (inside positive) generated with ISO vesicles was quantified by comparison to valinomycin-induced potassium diffusion potentials (Kout+→Kin+).
ΔpH (inside acidic) was measured with ISO vesicles by following the fluorescence quenching of 9-AA (20). The fluorescence was monitored by emission at 480 nm with excitation at 400 nm in the reaction mixture (1 ml total) consisting of 50 mM KPB (pH 6.5), 5 mM MgSO4, 5 μM 9-AA, and ISO membrane vesicles (≈100 μg of protein). Data were quantified by comparison to an artificial ΔpH established by suspending the vesicles (pH 6.0) in 50 mM KPB buffer of different pHs from 6.0 to 8.0.
Measurement of oxidase activity.Oxidase activity was measured using a Clark-type oxygen electrode at 25°C with 10 mM lactate, 10 mM succinate, or 0.5 mM NADH as the substrate. The reaction mixture (0.6 ml total) contained ISO membrane vesicles, 50 mM KPB (pH 6.5), and substrate, and the reaction was started by the addition of substrate. One unit was defined as the amount of enzyme which reduced 1 natom of O per min.
Transport assay.For the transport assay with intact cells, cells were collected at the late exponential phase, washed once with 50 mM KPB (pH 6.5), and suspended in glycerol medium containing 1.0% acetic acid (pH 3.5) to a final protein concentration of about 1.0 mg/ml. For the assay with membrane vesicles, RSO or ISO membrane vesicles were suspended with 1 ml of 50 mM KPB (pH 6.5) containing 5 mM MgSO4 at a final protein concentration of 10 to 500 μg/ml. The reaction was started by adding acetic acid containing [1-14C]sodium acetate (8 μCi, 6.25 μCi/mmol) to a final concentration of 0.4 mM into 1 ml of the cell or membrane suspension. Samples of cells or membrane vesicles, 50 μl in volume, were withdrawn at intervals and filtered through 0.45-μm-pore-size cellulose-acetate filters. The filters were quickly washed with 3 ml of cold 50 mM KPB (pH 6.5). After each filter membrane was put into a vial and 5 ml of scintillator solution (ACS II Scintillation Cocktail, Amersham) was added, retained radioactivity was counted by a scintillation counter.
Protein content.Protein content was determined by the amido black method (15). Each sample was precipitated with trichloroacetic acid in the presence of sodium dodecyl sulfate, filtered on a Millipore membrane, and stained with amido black 10B. The protein-dye complex that formed was then extracted with eluant solution, and the absorbance of the extract was measured at 630 nm. Bovine serum albumin was used as a standard protein.
Acetic acid and acetate accumulation in cells of A. aceti IFO 3283.The most plausible mechanism for acetic acid resistance in acetic acid bacteria is the operation of an efflux pump in the cytoplasmic membrane which pumps out toxic acetic acid and enables the cell to grow in the presence of high concentrations of it. In fact, it has been observed that an A. aceti strain adapted to high levels of acetic acid exhibits lower intracellular acetate concentrations than does a nonadapted strain (18).
To confirm the findings, acetic acid/acetate transport in A. aceti IFO 3283 was examined using [1-14C]sodium acetate and intact cells. When the labeled acetate was added to the cell suspension containing 1% acetic acid, the pH of the suspension being held at 3.5, only marginal acetate accumulation was observed, but it was increased by the addition of either carbonylcyanide m-chlorophenylhydrazone (CCCP) or KCN and more by the addition of both inhibitors (data not shown). The results suggested that at such a lower external pH, acetate can be passively accumulated but is pumped out by an efflux system which is inhibited by CCCP and/or KCN. However, since the metabolic conversion of acetate may occur inside the cell, the results obtained with the intact cells may lead to some under- or overestimation of the ability to pump acetate out of the cell.
Preparation of membrane vesicles from A. aceti IFO 3283.To overcome the uncertainty of the results obtained using intact cells, we tried to measure acetate accumulation or uptake with membrane vesicles from A. aceti IFO 3283. As described in Materials and Methods, we prepared RSO and ISO membrane vesicles by osmotic lysis of spheroplasts and by passage through a French pressure cell at relatively low applied pressures. As shown in Fig. 1, the RSO and ISO membrane vesicles thus prepared generated a lactate-dependent Δψ of inside negative and inside positive, respectively. The Δψ generated in opposite directions in the two membrane vesicle types was completely dissipated by the addition of nigericin and valinomycin, 2 mM KCN, or 10 μM CCCP (Fig. 1). The magnitude of the Δψ was dependent on the concentration of the membrane vesicles but was saturated at high concentrations (data not shown for RSO membrane vesicles and see Fig. 2A for ISO membrane vesicles). In ISO membrane vesicles, Δψ generation was also induced by succinate or NADH as well as by lactate, suggesting that it is generated by respiration.
Membrane potential generation in RSO (A) and ISO (B) membrane vesicles of A. aceti IFO 3283. The reaction mixture contained RSO membrane vesicles (A: 38.5 μg of protein) or ISO membrane vesicles (B: 415 μg of protein) in 50 mM KPB (pH 6.5) containing 5 mM MgSO4. The reaction was started by the addition of 10 mM lactate, followed by the addition of 0.025 μM nigericin (Nig) and 1 μM valinomycin (Val) or 2 mM KCN. Ten micromolar CCCP was also added before the addition of lactate. Fluorescence quenching of diS-C3-(5) (A) and diBA-C2-(5) (B) was monitored as described in Materials and Methods.
Membrane potential generation with different respiratory substrates and quantification of lactate-dependent membrane potential in ISO membrane vesicles. (A) The reaction mixtures contained different concentrations of ISO membrane vesicles in 50 mM KPB (pH 6.5) containing 5 mM MgSO4 and 2 μM diBA-C2-(5). The reactions were started by the addition of 10 mM lactate (▪), 10 mM succinate (▴), 0.5 mM NADH (○), or 3 mM ATP (pH 7.0) (▵), followed by the addition of nigericin (Nig) and valinomycin. ΔF/F indicates the rate (%) of fluorescence quenched per total unit of fluorescence. (B) ISO membrane vesicles were washed twice with 50 mM sodium phosphate buffer (NaPB, pH 6.5) to replace KPB with NaPB. The ISO membrane vesicles (10 μl; 101 μg of protein) were used for valinomycin-induced potassium diffusion potential measurement (▪), as described in Materials and Methods, and also for lactate-dependent Δψ (□) without Nig (+Lactate) or with Nig (+Nig) in 1 ml of the same buffer system as A.
The ISO membrane vesicles exhibited lactate, succinate and NADH oxidase activities of 0.120, 0.079, and 0.063 units/mg, respectively, and the activity was correlated with the magnitude of the Δψ generated (Fig. 2A). Furthermore, ATP could also be used to energize the vesicles (Fig. 2A), suggesting that the ISO vesicles still retained F1-ATPase with at least some level of exposure to the outside of the membrane vesicles. In RSO membrane vesicles, NADH, a membrane-nonpermeating substrate, could not generate any Δψ (inside positive or inside negative) (data not shown) despite its ability to generate a relatively large Δψ, inside positive, in ISO membrane vesicles, as described. Thus, the RSO membrane vesicles seemed not to have any substantial contamination by ISO membrane vesicles. The ISO membrane vesicles also seemed not to be contaminated by RSO vesicles, because lactate-dependent generation of Δψ (inside negative) was not observed (data not shown).
The lactate-dependent Δψ (inside positive) generation with ISO membrane vesicles was quantified based on comparison with potassium diffusion potential measurements (Fig. 2B). As shown, the Δψ generated with lactate was estimated to be 82 mV and 95 mV without and with nigericin, respectively. Lactate-dependent ΔpH generation with the ISO membrane vesicles was also estimated by 9-AA fluorescence measurement (Fig. 3). As shown, ISO membrane vesicles generated a ΔpH (inside acidic) of 1.33 and 1.46 without and with valinomycin, respectively. These data suggest that ISO membrane vesicles could generate a proton motive force consisting of a Δψ of 82 mV and a ΔpH of 1.33, total proton motive force of 161 mV, and also that the addition of either nigericin or valinomycin to ISO membrane vesicles could not completely compensate for the ΔpH or Δψ, respectively, so that ISO membrane vesicles produced a proton motive force of 95 mV and 86 mV in the presence of nigericin and valinomycin, respectively.
ΔpH measurement in ISO membrane vesicles of A. aceti IFO 3283. 9-AA fluorescence was measured with ISO membrane vesicles (101 μg of protein) prepared with 50 mM KPB (pH 6.0) as described in Materials and Methods. (A) The reaction was started by the addition of ISO vesicles to 50 mM KPB (pH 8.0), followed by the addition of 1 μM valinomycin (Val) and 0.1 μM nigericin (Nig) as indicated. (B) The reaction was started by the addition of lactate to the assay mixture containing ISO vesicles in 50 mM KPB (pH 6.0). (C) The quenching data (▪) obtained as shown in A by suspending the vesicles (pH 6.0) in 50 mM KPB buffer of pH 6.0, 6.5, 7.0, 7.5, and 8.0 were plotted against ΔpH, and lactate-dependent quenching data (□) without Val (+Lactate) or with Val (+Val) obtained in B were also plotted.
Acetic acid and acetate accumulation in RSO membrane vesicles.Accumulation of acetic acid and acetate in RSO membrane vesicles was examined. In contrast to intact cells, RSO membrane vesicles accumulated a relatively large amount of acetic acid/acetate, which was higher in the presence of a higher ΔpH between the vesicle inside (pH 6.5 or 5.0) and outside (pH 5.0 or 3.5) (Fig. 4). In spite of the presence or absence of a ΔpH, accumulation was almost completely eliminated by addition of the respiratory substrate lactate. Thus, as occurred with the intact cells having endogenous respiration, RSO membrane vesicles inhibited acetic acid/acetate accumulation as long as respiration was functioning.
Accumulation of acetic acid or acetate in RSO membrane vesicles of A. aceti IFO 3283. Accumulation of [1-14C]acetate was measured, as described in Materials and Methods, in the absence (○) and the presence (▪) of 10 mM lactate. The reactions were performed with RSO membrane vesicles (38.5 μg of protein) prepared with 50 mM KPB (pH 6.5) in McIlvaine buffer (pH 3.5) (A), in 50 mM MacIlvaine buffer (pH 5.0) (B), and also with vesicles prepared with 50 mM KPB (pH 5.0) in 50 mM KPB (pH 5.0) (C).
Acetic acid and acetate uptake in ISO membrane vesicles.Uptake of acetic acid and acetate into ISO membrane vesicles was also examined. There was a weak acetic acid/acetate uptake into the vesicles when no substrate was present, but the uptake was stimulated in the presence of the respiratory substrate lactate, and this activity was completely blocked by the addition of CCCP (Fig. 5 and 6). As shown in Fig. 5A, the lactate-dependent acetic acid/acetate uptake was partially inhibited by either nigericin or valinomycin and completely inhibited by both ionophores together. The partial inhibition seems to be related to the findings that conversion of Δψ or ΔpH was not fully achieved upon addition of valinomycin or nigericin, respectively (Fig. 2 and 3). Furthermore, acetic acid/acetate uptake was observed after the addition of ATP instead of a respiratory substrate, although the intensity was lower than that for lactate (Fig. 5B). The ATP-dependent uptake was inhibited by dicyclohexylcarbodiimide, an inhibitor of F0-ATPase, indicating that it also occurs via a proton motive force generated by ATP hydrolysis with ATPase but not by ATP itself.
Acetic acid or acetate uptake in ISO membrane vesicles of A. aceti IFO 3283. The transport of [1-14C]acetate was measured with ISO membrane vesicles at 537 μg (A) or 540 μg (B) of protein in 50 mM KPB (pH 6.5) containing 5 mM MgSO4, as described in Materials and Methods. When indicated, 0.025 μM nigericin (Nig), 1 μM valinomycin (Val), 10 μM CCCP, or 100 μM DCCD was added to the ISO membrane vesicles suspension and incubated before the addition of [1-14C]acetate. (A) The reaction was carried out in the absence (•) or presence of 10 mM lactate (□), lactate and Nig (▴), lactate and Val (○), or lactate, Nig and Val (▵). (B) The reaction was done in the absence (•) or presence of 3 mM ATP (□), ATP and CCCP (○), or ATP and DCCD (▵).
Effect of pH on acetic acid or acetate uptake in ISO membrane vesicles of A. aceti IFO 3283. The reactions were carried out using ISO membrane vesicles (545 μg of protein) as described in Materials and Methods. (A) The reaction was carried out in 50 mM MacIlvaine buffer (pH 5.0) in the absence of lactate and CCCP (•) or in the presence of 10 mM lactate (▪) or 10 mM lactate and 10 μM CCCP (▵). (B) The reactions were carried out under different buffer conditions (MacIlvaine buffer at pH 5.0 and 5.5 and 50 mM KPB at pH 6.0, 6.5, and 7.0) with lactate (▪) and with lactate and CCCP (▵) and also in the absence of the substrate and the inhibitor (•).
Substrate specificity in acetic acid and acetate uptake of ISO membrane vesicles.There is some uncertainty whether acetic acid or acetate is the substrate for the efflux pump, because sodium acetate equilibrates between protonated acetic acid and deprotonated acetate in solution. Since the ratio of the two species is pH dependent, we may expect which species is the substrate for the pump by examining uptake at different pHs. As can be compared in Fig. 5A and Fig. 6A, uptake was largely increased at pH 5 relative to pH 6.5. And also, as shown in Fig. 6B, there was a clear pH dependence for acetic acid/acetate uptake in ISO membrane vesicles, being higher at lower pH. Although the passive diffusion of acetic acid that occurs without a respiratory substrate was also increased by decreasing the pH outside, the uptake of acetic acid was very much increased by the addition of lactate, and the increased acetic acid accumulation also exhibited pH dependence.
To further characterize the substrate specificity, acetic acid/acetate uptake in ISO membrane vesicles was examined in the presence of 2.5- and 50-fold molar excesses of other substrates, such as propionic acid, butyric acid, sorbic acid, and ethanol (Fig. 7). All these compounds could dissipate the Δψ generated in RSO membrane vesicles at a millimolar concentration except for propionate, which does so at a concentration of 200 mM or more, like acetic acid (data not shown). Thus, the uptake experiments were performed with 20 μM and 400 μM acetate in the presence of 1 mM of other acids or ethanol. As shown, propionic acid and butyric acid inhibited acetic acid/acetate uptake a little at a 50-fold molar excess but not at a 2.5-fold molar excess. Sorbic acid and ethanol did not have any effect on uptake, although sorbic acid had some inhibitory activity by exhibiting some uncoupling ability.
Effect of other substrates on acetic acid or acetate uptake in ISO membrane vesicles of A. aceti IFO 3283. By adding acetic acid containing [1-14C]sodium acetate to a final concentration of 20 μM (open bar) or 0.4 mM (solid bar), the transport assay was carried out at pH 5.0 with ISO membrane vesicles (380 μg of protein) in the absence (control) or presence of 1 mM propionate, butyrate, sorbate, or ethanol.
Acetic acid resistance is a crucial ability allowing acetic acid bacteria to stably produce large amounts of acetic acid. In Acetobacter and Gluconacetobacter species, spontaneous mutations occur at high frequencies and sometimes lead to simultaneous defects in their acetate-producing (ethanol-oxidizing) and acetic acid resistance abilities (11, 19). Furthermore, alcohol dehydrogenase (ADH) disruption in Acetobacter pasteurianus has also been shown to lead to loss of acetic acid resistance (1). Since ADH deficiency causes defects in ethanol respiration, loss of acetic acid resistance may be related to respiration, from which a proton motive force is generated, via ubiquinol oxidase (9) (see Fig. 8B). Therefore, if the acetic acid resistance mechanism requires energy, ethanol respiration and the energy it produces may be important. An energy-requiring mechanism responsible for acetic acid resistance is expected to be an efflux pump in the cytoplasmic membrane which pumps out acetic acid and enables the cell to grow in the presence of high concentrations of acetic acid. For example, Pdr12 in Saccharomyces cerevisiae acts as an ATP-dependent acetate (but not acetic acid) efflux pump, allowing the cell to adapt to high acetic acid concentrations (4, 12).
Schematic representation of acetic acid uptake in ISO membrane vesicles and model of the efflux pump present in the cytoplasmic membrane of A. aceti. (A) ISO membrane vesicles generate a proton motive force, inside positive and acidic, by respiration (lactate oxidation), which then drives acetic acid uptake. The stoichiometry of protons and acetic acid in the pump is unknown. There is also a passive diffusion of acetic acid, which is ΔpH dependent. (B) In intact cells, ethanol respiration produces acetic acid from ethanol via the activity of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), which also donate electrons to a terminal ubiquinol oxidase (Ox) that in turn generates a proton motive force. The acetic acid produced can passively penetrate into the cytoplasm but is pumped out by an efflux pump which is energized by the proton motive force.
To examine this hypothesis, we first measured acetic acid and acetate accumulation in intact cells and in RSO membrane vesicles of A. aceti IFO 3283. Acetate accumulation was low in intact cells having endogenous respiration, but it was increased by the addition of protonophores and/or a respiratory inhibitor. In contrast, RSO membrane vesicles accumulated a relatively high concentration of acetic acid in a ΔpH-dependent passive diffusion manner, but accumulation was decreased almost completely by respiration. The results suggest that cells or RSO membrane vesicles actively pump out acetic acid which has passively penetrated into the cell by using energy produced by respiration. Such an acetic acid efflux mechanism in A. aceti was also demonstrated using ISO membrane vesicles. The ISO membrane vesicles, able to generate a large enough inside-positive Δψ by respiration, could accumulate acetic acid/acetate in the presence of respiratory substrates in a proton uncoupler-sensitive manner (Fig. 8A). The results obtained for the vesicles also indicated that the energy source required for uptake is the proton motive force but not ATP (Fig. 5).
Another question associated with the efflux pump was the identity of the substrate for this transporter, since the acetate anion is expected to be a substrate for the Pdr12 efflux pump in yeast (4, 12). In ISO membrane vesicles of A. aceti, lactate-dependent acetic acid/acetate uptake was proportionally increased by decreasing the pH (Fig. 6). Since lactate oxidase activity has an optimum pH of 6.5, the higher uptake at lower pH seems not to be related to the degree of energy generation. Although it is not definitive evidence, this result suggests that acetic acid but not acetate is transported inside the vesicles. It also suggested that the efflux pump is relatively specific for acetic acid, though it may work for high concentrations of short-chain acids such as propionic acid and butyric acid (Fig. 7).
The efflux pump may not work for ethanol, although Acetobacter species also exhibit resistance to high concentrations of alcohol. In yeast cells, acetic acid is expected to be pumped out by coupling the Pdr12 ABC transporter, which transports acetate anions, with the Pma1 plasma membrane H+-ATPase (12). However, acetic acid bacteria such as A. aceti seem not to have any proton efflux system such as Pma1, and the acetic acid efflux system we observed seems to be an H+-antiporter but not an ABC transporter (Fig. 8A). Thus, it is reasonable to expect that the efflux system of acetic acid bacteria is pumping out "protonated" acetic acid, much the same as a proton motive force-dependent multidrug transporter.
The results obtained in this study therefore suggest that a proton motive force-dependent acetic acid efflux pump exists in A. aceti IFO 3283. Although cells grown in glycerol medium were used in this study, much higher levels of efflux activity are expected to occur in ethanol culture, where Acetobacter species produce and accumulate a large amount of acetic acid outside of the cell and thus cannot survive in such a culture without machinery able to eliminate acetic acid. Therefore, as shown in Fig. 8B, the acetic acid efflux pump seems to be highly expressed and expels acetic acid by using the proton motive force produced by ethanol respiration. We have found much higher activity of the efflux pump in cells in the ethanol culture compared to the glycerol culture used in this study (unpublished observation). Now, we are examining the activity change of the efflux pump in the different growth phases of ethanol cultures of A. aceti IFO 3283.
Received 28 October 2004.
Accepted 29 March 2005.
Chinnawirotpisan, P., G. Theeragool, S. Limtong, H. Toyama, O. Adachi, and K. Matsushita. 2003. Quinoprotein alcohol dehydrogenase is involved in catabolic acetate production, while NAD-dependent alcohol dehydrogenase in ethanol assimilation in Acetobacter pasteurianus SKU1108. J. Biosci. Bioeng.96:564-571.
Fukaya, M., H. Takemura, H. Okumura, Y. Kawamura, S. Horinouchi, and T. Beppu. 1990. Cloning of genes responsible for acetic acid resistance in Acetobacter aceti. J. Bacteriol.172:2096-2104.
Fukaya, M., H. Takemura, K. Tayama, H. Okumura, Y. Kawamura, S. Horinouchi, and T. Beppu. 1993. The aarC gene responsible for acetic acid assimilation confers acetic acid resistance on Acetobacter aceti. J. Ferment. Bioeng.76:270-275.
Hatzixanthis, K., M. Mollapour, L. Seymour, B. E. Bauer, G. Krapf, C. Schuller, K. Kuchler, and P. W. Piper. 2003. Moderately lipophilic carboxylate compounds are the selective inducers of the Saccharomyces cerevisiae Pdr12p ATP-binding cassette transporter. Yeast20:575-585.
Lasko, D. R., C. Schwerdel, J. E. Bailey, and U. Sauer. 1997. Acetate-specific stress response in acetate-resistant bacteria: an analysis of protein patterns. Biotechnol. Prog.13:519-523.
Matsushita, K., and M. Ameyama. 1982. d-Glucose dehydrogenase from Pseudomonas fluorescens, membrane-bound. Methods Enzymol.89:149-153.
Matsushita, K., M. Nonobe, E. Shinagawa, O. Adachi, and M. Ameyama. 1985. Isolation and characterization of outer and cytoplasmic membranes from spheroplasts of Acetobacter aceti. Agric. Biol. Chem.49:3519-3526.
Matsushita, K., and H. R. Kaback. 1986. d-Lactate oxidation and generation of the proton electrochemical gradient in membrane vesicles from Escherichia coli GR19N and in proteoliposomes reconstituted with purified d-lactate dehydrogenase and cytochrome o oxidase. Biochemistry25:2321-2327.
Matsushita, K., H. Toyama, and O. Adachi. 1994. Respiratory chain and bioenergetics of acetic acid bacteria. Adv. Microb. Physiol.36:247-301.
Nakano, S., M. Fukaya, and S. Horinouchi. 2004. Enhanced expression of aconitase raises acetic acid resistance in Acetobacter aceti. FEMS Microbiol. Lett.235:315-322.
Ohmori, S., T. Uozumi, and T. Beppu. 1982. Loss of acetic acid resistance and ethanol oxidizing ability in an Acetobacter strain. Agric. Biol. Chem.46:381-389.
Piper, P., C. O. Calderon, K. Hatzixanthis, and M. Mollapour. 2001. Weak acid adaptation: the stress response that confers yeasts with resistance to organic acid food preservatives. Microbiology147:2635-2642.
Saeki, A., M. Taniguchi, K. Matsushita, H. Toyama, G. Theeragool, N. Lotong, and O. Adachi. 1997. Microbiological aspects of acetate oxidation by acetic acid bacteria, unfavorable phenomena in vinegar fermentation. Biosci. Biotechnol. Biochem.61:317-323.
Saeki, A., K. Matsushita, S. Takeno, M. Taniguchi, H. Toyama, G. Theeragool, N. Lotong, and O. Adachi. 1999. Enzymes responsible for acetate oxidation by acetic acid bacteria. Biosci. Biotechnol. Biochem.63:2102-2109.
Schaffner, W., and C. Weissman. 1973. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal. Biochem.56:502-514.
Sievers, M., C. Gaberthuel, C. Boesch, W. Ludwig, and M. Teuber. 1995. Phylogenetic position of Gluconobacter species as a coherent cluster separated from all Acetobacter species on the basis of 16S ribosomal RNA sequences. FEMS Microbiol. Lett.126:123-126.
Steiner, P., and U. Sauer. 2001. Proteins induced during adaptation of Acetobacter aceti to high acetate concentrations. Appl. Environ. Microbiol.67:5474-5481.
Steiner, P., and U. Sauer. 2003. Long-term continuous evolution of acetate resistant Acetobacter aceti. Biotechnol. Bioeng.84:40-44.
Takemura, H., S. Horinouchi, and T. Beppu. 1991. Novel insertion sequence IS1380 from Acetobacter pasteurianus is involved in loss of ethanol-oxidizing ability. J. Bacteriol.173:7070-7076.
Toyomizu, M., K. Okamoto, Y. Akiba, T. Nakatsu, and T. Konishi. 2002. Anacardic acid-mediated changes in membrane potential and pH gradient across liposomal membranes. Biochim. Biophys. Acta1558:54-62.
Yamada, Y., K. Hoshino, and T. Ishikawa. 1997. The phylogeny of acetic acid bacteria based on the partial sequences of 16S ribosomal RNA: The elevation of subgenus Gluconoacetobacter to the generic level. Biosci. Biotechnol. Biochem.61:1244-1251.
Yamada, Y., K. Katsura, H. Kawasaki, Y. Widyastuti, S. Saono, T. Seki, T. Uchimura, and K. Komagata. 2000. Asaia bogorensis gen. nov., sp. nov., an unusual acetic acid bacterium in the alpha-Proteobacteria. Int. J. Evol. Microbiol.50:823-829.
Journal of Bacteriology Jun 2005, 187 (13) 4346-4352; DOI: 10.1128/JB.187.13.4346-4352.2005
Thank you for sharing this Journal of Bacteriology article.
You are going to email the following Acetobacter aceti Possesses a Proton Motive Force-Dependent Efflux System for Acetic Acid
Message Subject (Your Name) has forwarded a page to you from Journal of Bacteriology
Message Body (Your Name) thought you would be interested in this article in Journal of Bacteriology.
Acetobacter
Proton-Motive Force
Follow #Jbacteriology


Climb inside the 2018 Jeep Wrangler JK. With less than 2,000 miles on the odometer, this 4 door sport utility vehicle prioritizes comfort, safety and convenience. It features four-wheel drive capabilities, a durable automatic transmission, and a refined 6 cylinder engine. Top features include power windows, a tachometer, a trip computer, an automatic dimming rear-view mirror, front fog lights, skid plates, a trailer hitch, and voice activated navigation. Features such as automatic climate control and leather upholstery prove that economical transportation does not need to be sparsely equipped. Premium sound drives 9 speakers, providing you and your passengers a sensational audio experience. Jeep also prioritized safety and security with features such as: dual front impact airbags, integrated roll-over protection, traction control, brake assist, a security system, and 4 wheel disc brakes with ABS. With electronic stability control supplementing mechanical systems, you'll maintain precise command of the roadway. A Carfax history report indicates just one previous owner. Our sales reps are extremely helpful & knowledgeable. They'll work with you to find the right vehicle at a price you can afford. Please don't hesitate to give us a call.


Are either of these issues already being worked on or need idea for them?
Another item that was not fully updated with Winter 19 is the Theme editor. If y ou look at the screenshot at the bottom or the pop-up on hover image, the UI shows the new logo with "Helix" but still has the old activity icon. This is very minor but wanted to mention it.
We too like that the text is no longer orange but maybe a darker font color would be better. Would be nice to be able to customize these to fit our branding.
Thanks for pointing out the error in the theme editor.
The Drafts counter background color is not hard-coded to orange; it is the active button color for the theme. Introducing a new configuration just for this purpose was not felt necessary in the interest of keeping it simple.
I guess I didn't apply my theme after updating it so you're right, the counter fill color changes. But the OUTLINE color is still orange. I have no orange in my theme so it must be hard-coded to be that way? Defect or Idea?
You also didnt answer whether the "Looking for" text can be darkened to be the same color as "View in My Activity". Defect or Idea?
Defect no. 94365 has been created to fix the outline.


Newspapers provide news and editorials. It is the editorials that slant the truth to the opinions of the owners. The same is true of television where there are news shows and there are opinion shows.
When we watch these shows we need to remember which ones we are watching. We can be very gullible and opinion shows take advantage of the gullible person.


text:
//
// NotificationPageBuilder.cs
//
// Author:
//       Seyed Razavi <monkeyx@gmail.com>
//
// Copyright (c) 2015 Seyed Razavi
//
// Permission is hereby granted, free of charge, to any person obtaining a copy
// of this software and associated documentation files (the "Software"), to deal
// in the Software without restriction, including without limitation the rights
// to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
// copies of the Software, and to permit persons to whom the Software is
// furnished to do so, subject to the following conditions:
//
// The above copyright notice and this permission notice shall be included in
// all copies or substantial portions of the Software.
//
// THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
// IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
// FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE
// AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
// LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,
// OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN
// THE SOFTWARE.
using System;
using System.Collections.Generic;

using Xamarin.Forms;

using Phoenix.BL.Entities;
using Phoenix.BL.Managers;

namespace PhoenixImperator.Pages.Entities
{
	/// <summary>
	/// Notification page builder.
	/// </summary>
	public class NotificationPageBuilder : BaseEntityPageBuilder<Notification>
	{
		/// <summary>
		/// Initializes a new instance of the <see cref="PhoenixImperator.Pages.Entities.NotificationPageBuilder"/> class.
		/// </summary>
		public NotificationPageBuilder ()
		{
		}

		/// <summary>
		/// Displaies the entity.
		/// </summary>
		/// <param name="item">Item.</param>
		protected override void DisplayEntity (Notification item)
		{
			entityPage.Title = item.TypeDescription;

			string dateLabel;
			if (item.DaysAgo > 0) {
				if (item.DaysAgo == 1) {
					dateLabel = ("Yesterday");
				} else {
					dateLabel = (item.DaysAgo + " days ago");
				}
			} else {
				dateLabel = ("Today");
			}

			AddContentTab (dateLabel, "icon_notifications.png");
			currentTab.AddHeading (item.ListText);

			if (item.Type == Notification.NotificationType.ComplexChange) {
				currentTab.AddLabel (item.ChangeType.ToString ());
			}

			if (item.Type == Notification.NotificationType.RegisteredBaseActivity) {
				currentTab.AddLabel (item.BaseActivityType.ToString ());
			}

			// AddProperty ("Priority", item.Priority.ToString ());

			if (item.Position != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.Position, "Position");
			} else if(!string.IsNullOrWhiteSpace(item.PositionName)){
				currentTab.AddPropertyDoubleLine ("Position", item.PositionName);
			}

			if (item.StarSystem != null) {
				currentTab.AddEntityProperty (Phoenix.Application.StarSystemManager, item.StarSystem, "Star System");
			} else if(!string.IsNullOrWhiteSpace(item.StarSystemName)){
				currentTab.AddPropertyDoubleLine ("Star System", item.StarSystemName);
			}

			if(!string.IsNullOrWhiteSpace(item.CelestialBodyName)){
				currentTab.AddPropertyDoubleLine ("Celestial Body", item.CelestialBodyName);
			}

			if (item.Order != null) {
				currentTab.AddEntityProperty (Phoenix.Application.OrderTypeManager, item.Order, "Order");
			} else if(!string.IsNullOrWhiteSpace(item.OrderName)){
				currentTab.AddPropertyDoubleLine ("Order", item.OrderName);
			}

			if (item.PositionType != Notification.NotificationPositionType.None) {
				currentTab.AddProperty ("Position Type", item.PositionType.ToString ());
			}

			if(!string.IsNullOrWhiteSpace(item.SquadronName)){
				currentTab.AddPropertyDoubleLine ("Squadron", item.SquadronName);
			}

			if(!string.IsNullOrWhiteSpace(item.Location)){
				currentTab.AddPropertyDoubleLine ("Location", item.Location);
			}

			if (item.NumberOfShips > 0) {
				currentTab.AddProperty ("Ships", item.NumberOfShips.ToString ());
			}

			if (item.Quantity > 0) {
				currentTab.AddProperty ("Quantity", item.Quantity.ToString ());
			}

			if (item.Item != null) {
				currentTab.AddEntityProperty (Phoenix.Application.ItemManager, item.Item, "Item");
			} else if(!string.IsNullOrWhiteSpace(item.ItemName)){
				currentTab.AddPropertyDoubleLine ("Item", item.ItemName);
			}

			if (item.Base != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.Base, "Base");
			} else if(!string.IsNullOrWhiteSpace(item.BaseName)){
				currentTab.AddPropertyDoubleLine ("Base", item.BaseName);
			}

			if (item.DeliverTo != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.DeliverTo, "Delivered To");
			} else if(!string.IsNullOrWhiteSpace(item.DeliverToName)){
				currentTab.AddPropertyDoubleLine ("Deliver To", item.DeliverToName);
			}

			if (item.PickedUpFrom != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.PickedUpFrom, "Picked Up From");
			} else if(!string.IsNullOrWhiteSpace(item.PickedUpFromName)){
				currentTab.AddPropertyDoubleLine ("Picked Up From", item.PickedUpFromName);
			}

			if (item.BoughtFrom != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.BoughtFrom, "Bought From");
			} else if(!string.IsNullOrWhiteSpace(item.BoughtFromName)){
				currentTab.AddPropertyDoubleLine ("Bought From", item.BoughtFromName);
			}

			if (item.SoldTo != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.SoldTo, "Sold To");
			} else if(!string.IsNullOrWhiteSpace(item.SoldToName)){
				currentTab.AddPropertyDoubleLine ("Sold To", item.SoldToName);
			}

			if (item.ByPosition != null) {
				currentTab.AddEntityProperty (Phoenix.Application.PositionManager, item.ByPosition, "By");
			} else if(!string.IsNullOrWhiteSpace(item.ByPositionName)){
				currentTab.AddPropertyDoubleLine ("By", item.ByPositionName);
			}

			if (item.Stellars > 0) {
				currentTab.AddProperty ("Stellars", item.Stellars.ToString ());
			}

			if (!string.IsNullOrWhiteSpace (item.AffiliationName)) {
				currentTab.AddPropertyDoubleLine ("Affiliation", item.AffiliationName);
			}

			if (!string.IsNullOrWhiteSpace (item.Status)) {
				currentTab.AddPropertyDoubleLine ("Status", item.Status);
			}

			if (item.Type == Notification.NotificationType.PlanetarySales) {
				currentTab.AddProperty ("Planetary Sales", item.TradeType.ToString ());
			}

			if (!string.IsNullOrWhiteSpace (item.Subject)) {
				currentTab.AddPropertyDoubleLine ("Subject", item.Subject);
			}

			if (!string.IsNullOrWhiteSpace (item.Message)) {
				currentTab.AddPropertyDoubleLine ("Message", item.Message);
			}

			if (!string.IsNullOrWhiteSpace (item.ErrorCodeDescription)) {
				currentTab.AddProperty ("Error Code", item.ErrorCodeDescription);
			}

			if (!string.IsNullOrWhiteSpace (item.ErrorMessage)) {
				currentTab.AddPropertyDoubleLine ("Message", item.ErrorMessage);
			}

			if (!string.IsNullOrWhiteSpace (item.WarningCodeDescription)) {
				currentTab.AddProperty ("Warning Code", item.WarningCodeDescription);
			}

			if (!string.IsNullOrWhiteSpace (item.WarningMessage)) {
				currentTab.AddPropertyDoubleLine ("Message", item.WarningMessage);
			}

			if (item.ComplexItem != null) {
				currentTab.AddEntityProperty (Phoenix.Application.ItemManager, item.ComplexItem, "Complex");
			} else if(!string.IsNullOrWhiteSpace(item.ComplexTypeName)){
				currentTab.AddPropertyDoubleLine ("Complex", item.ComplexTypeName);
			}
		}
	}
}

redpajama_set_name: RedPajamaGithub


We cover Access Risk Analysis, Emergency Access Management, Access Request Management , Business Role Management and Migration from GRC 5.3 to GRC 10.0. Below is the course content for GRC 10.0 online Training. Enquire here for more details.
instructors providing real time courses in our New Batch starts every week.
We proud to say more than 90% of students have got placed after the training.
Very Experienced Quality Faculties who have more than 15 years hands on experience.
SAP GRC online training offer you easy way to learn the SAP GRC course.


text: Self made personal planner, featuring a calendar.