text:
Work force
In Globe's musical ode to toil, the unsung take a bow
Why it's work and why it's fun
WORKER: Gordon Greenberg
PRESENT POSITION: Director
FIRST JOB: At age 12, cast as a newsboy in ... "Working."
WORST JOB: Producer and director of commercials for an ad agency. It wasn't so much the job as the setting: Greenberg recalls sitting down in his cubicle, looking up at the fluorescent lights "and thinking, 'I'm going to spend the bulk of my life here. This is it.' I had a little panic attack. I took the elevator down and walked around the block about five times and breathed, and got through it."
WHY THIS IS WORK: "I think it's a well-known fact it's not an easy way to make a living. Nor is it the most lucrative. They used to say you can't make a living in the theater, but you can make a killing. The challenging parts are being away from home – you miss your friends, your relationship and your family. You are constantly jockeying to keep your projects alive. Making sure they're still relevant, artistically and commercially. And making sure you're keeping everyone happy."
WHY THIS IS WORTH IT: "I love telling stories, and I love seeing the power of storytelling with an audience, and the power of being a part of a big group and experiencing something together. Which is why theater is so essential."
WORKER: Lin-Manuel Miranda
PRESENT POSITION: Co-composer
FIRST JOB: Cashier and delivery boy at McDonald's, 90th Street and Columbus Avenue in Manhattan.
WORST JOB: That cashier position. "The register was just a miserable, miserable experience. But then deliveries were just, like, freedom. I felt untethered from the world."
WHY THIS IS WORK: After a year performing the taxing lead role of Usnavi in the Broadway hit "In the Heights," Miranda looked forward to "going back to being a writer, and not having physical therapy once a week." As a composer, Miranda works way, way overtime: he cut some 60 songs from "In the Heights" in the time between its conception (when he was still a student at Wesleyan College) and its Broadway debut.
WHY THIS IS WORTH IT: "Having a musical is like having a baby. We're the village that raised it, and we're going to take care of it for the rest of its life." And teaming with Schwartz on "Working" is "very exciting, because it's being let into the club. It's an incredible group of songwriters who got to work on that (original) show."
WORKER: Marie-France Arcilla
PRESENT POSITION: Performer
FIRST JOB: At age 8, anchored the "Children's News Hour" for a TV station in Manila, Philippines (her home country), for $6 an episode. After she moved to the States six years ago, sold bread at the Brooklyn Greenmarket.
WORST JOB: "I've been fortunate enough that if maybe the job wasn't that interesting, at least the people were nice. But when I was working coat check in a restaurant, it was kind of what they talk about in the show – the lack of recognition. People kind of glaze over, they don't see you. One time, a drunk guy took the coat tag with him (which she needed to keep). I ran after him for the tag, and a guy at the bar said, 'Yeah baby, run after that dollar.'
"Now I tip well."
WHY THIS IS WORK: "Sometimes my full-time job is looking for a job. That's my life." Arcilla notes that castmate Wayne Duvall has done a lot of film and TV work, and they've talked about how "what he does is there for all to see. It's on DVD. It's on YouTube. And for the most part, what we do in the theater, once it's done – that particular night, that particular performance – it's gone."
WHY THIS IS WORTH IT: "What's wonderful about it is it's fresh all the time. It's not set in stone. One performance is not going to define me. (And) I feel incredibly satisfied in the theater when there's a young kid there, or a grandmother from New Jersey, and she says, 'You really touched my life.' "
WORKER: Mark Hartman
PRESENT POSITION: Music director/conductor
FIRST JOB: Playing organ and piano at age 12 in the rural North Carolina Baptist church where his father was a minister.
WORST JOB: Repackaging cigarettes at an R.J. Reynolds Tobacco Co. factory at age 17. He lasted one day, then got a new job at Action Video.
WHY THIS IS WORK: "Well, my parents still don't think I really have a job," says Hartman, who is on leave as music director of the hit "Avenue Q" on Broadway. " 'You do what?' But there are similarities to every other job on the planet. It's a group of people getting together to make a product. What we produce happens to be a play."
WHY THIS IS WORTH IT: "The most satisfying thing in rehearsals is working with actors to create a moment out of a song that is sculpted specifically for them. Taking a song on a page, and finding a key or an arrangement or a moment in the song that allows them to shine both as singers and actors." Plus, "When I'm the conductor in the pit and there's an orchestra, that's a stunning feeling."
WORKER: Dan Rosokoff
PRESENT POSITION: Stage manager
FIRST JOB: Stocking shelves and working the register at the pharmacy his dad co-owned in Tonawanda, NY.
WORST JOB: Temping as a personnel assistant at the National Heart, Lung and Blood Institute in Bethesda, Md. The verdict: "Boooorring. It was monotonous. It was just the same thing driving to work every day." Rosokoff adds that he's not belittling the job: "It just wasn't right for me."
WHY THIS IS WORK: No matter how much you plan, things can and do go wrong. Like the time during a production of "Jesus Christ Superstar" that a wick flew out of a lit torch and started a fire onstage. "Here I am, very young, thinking 'Oh my God, I'm the stage manager, I'm in charge,' " he recalls. "I grabbed a fire extinguisher off the wall, walked onstage and put out the fire. We cleaned up the stage and went on with the show."
WHY THIS IS WORTH IT: "When you're working on a new show, just seeing all the creativity in the room – whether it's a playwright or composer or director or choreographer, with all their wants, wishes, dreams – listening to all that, and then imparting it to the other people who make it happen. And when that exciting moment happens onstage with an audience for the first time, you get to remember the moment someone came up with the idea."


text:
This white paper discusses how you can transform your customer
service appropriately-and-cost-effectively-to move toward delighting people.
A new white paper by the 451 Research Mobility Team (powered by Yankee Group) addresses opportunity to improve customer service via mobile.
Smart phones and tablets are now universal tools providing anywhere-anytime communication. These mobile devices empower managers to monitor and respond to changing contact center operations in real time. Forward-looking contact centers can use such advanced technology to better manage operations and service customer needs. This whitepaper explores advantages of using an integrated workforce management system with mobile devices to attain superior efficiency at less cost.
Most companies get mobile self-service all wrong, and frustrate customers as a result. This new report shows you how to finally get the service experience right in this popular channel.

subset: commoncrawl


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- Mark's Daily Apple - http://www.marksdailyapple.com -
Weekend Link Love – Edition 143
Posted By Mark Sisson On May 22, 2011 @ 5:00 am In Weekend Link Love | 41 Comments
Did you know Denmark has levied a tax on saturated fat [7], with Finland considering enacting a similar tax? Shameful. The vikings are rolling in their funeral pyres.
The EU is banning certain herbal remedies [8]. I wonder what else is next – and where?
For pro athletes, positive tests are usually bad news. Not Novak Djokovic; the number two tennis player in the world credits much of his success [9] to giving up "bread, pasta, and pizza" after testing positive for gluten sensitivity.
Political scientist James Scott has been delivering a series of lectures on the domestication of man via grain and government [10]. Though Scott's aren't available for download yet, hundreds from previous lecture series are available [11].
A study of US college students [12] found that children of Asian immigrants were likely to avoid eating traditional foods – feet, eyes, bones, and faces – out of embarrassment and a desire to fit in.
Food producers overhype and exaggerate [13] their "functional food" products' health benefits. I can't say I'm surprised… remember magic yogurt [14]?
Primal gamers and LARPers might consider joining the upcoming beta for City of Epic [15], a new browser-based exercise RPG where doing real-world workouts earns you in-game points and power-ups. Here's a shorter video [16].
Boing Boing highlights a sweet, stylish stand-up workstation [17].
And finally, how to chop wood [18] without messing around.
One year ago (May 17 – May 23)
I've begun telling people, "Alcohol is an even more preferred fuel than glucose. So when are you switching to a beertarian diet?"
– Commenter Dana, from earlier this week [23].
Article printed from Mark's Daily Apple: http://www.marksdailyapple.com
URL to article: http://www.marksdailyapple.com/weekend-link-love-143/
URLs in this post:
[1] Start Here: http://www.marksdailyapple.com/welcome-to-marks-daily-apple/?utm_source=mda_wwsgd&utm_medium=link&utm_campaign=mda_wwsgd_start_here
[2] Primal Blueprint 101: http://www.marksdailyapple.com/primal-blueprint-101/?utm_source=mda_wwsgd&utm_medium=link&utm_campaign=mda_wwsgd_pb_101
[3] free weekly newsletter: http://www.marksdailyapple.com/subscribe-to-blog/?utm_source=mda_wwsgd&utm_medium=link&utm_campaign=mda_wwsgd_newsletter
[4] books: http://primalblueprint.com/categories/Store/Books-and-Media/?utm_source=mda_wwsgd&utm_medium=link&utm_campaign=mda_wwsgd_books
[5] support options: http://primalblueprint.com/categories/Store/Services/?utm_source=mda_wwsgd&utm_medium=link&utm_campaign=mda_wwsgd_services
[6] supplements: http://primalblueprint.com/categories/Store/Supplements/?utm_source=mda_wwsgd&utm_medium=link&utm_campaign=mda_wwsgd_supplements
[7] tax on saturated fat: http://www.just-food.com/analysis/denmarks-saturated-fat-tax-provokes-industry-anger_id115300.aspx
[8] banning certain herbal remedies: http://www.naturalnews.com/032389_herbal_remedies_Europe.html
[9] credits much of his success: http://www.10sballs.com/2011/04/26/djokovic-ditches-pizza-to-push-for-top-spot/
[10] domestication of man via grain and government: http://news.harvard.edu/gazette/story/2011/05/tanner-lecture/?sms_ss=email&at_xt=4dc40b5d1e0f10e0,0
[11] are available: http://www.tannerlectures.utah.edu/lectures/atoz.html
[12] study of US college students: http://www.livescience.com/14143-supersized-immigrants.html?utm_source=feedburner&utm_medium=feed&utm_campaign=Feed:+Livesciencecom+(LiveScience.com+Science+Headline+Feed)
[13] overhype and exaggerate: http://www.nytimes.com/2011/05/15/business/15food.html?_r=1
[14] magic yogurt: http://www.marksdailyapple.com/yogurt-mania/
[15] City of Epic: http://www.kickstarter.com/projects/1171791306/city-of-epic-an-exercise-rpg
[16] shorter video: http://www.youtube.com/watch?v=iT9eJfygwg4
[17] sweet, stylish stand-up workstation: http://www.boingboing.net/2011/05/17/standing-desk-jockey.html
[18] chop wood: http://www.wimp.com/chopwood/
[19] roast chicken and sex with your partner: http://www.slate.com/id/2294849
[20] how to cook his famous omelette: http://www.youtube.com/watch?v=487iOLwnrsU&feature=youtu.be&hd=1
[21] The Power of Touch: http://www.marksdailyapple.com/the-power-of-touch/
[22] Smart Spice: Cinnamon: http://www.marksdailyapple.com/health-benefits-cinnamon/
[23] earlier this week: http://www.marksdailyapple.com/a-metabolic-paradigm-shift-fat-carbs-human-body-metabolism/#comments
Click here to print.
Copyright © 2009 Mark's Daily Apple. All rights reserved.


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Vendôme, France Bed & Breakfasts - Set amidst gentle hillsides, two large suites with private baths and exquisite views on the valley. 20 miles from Blois, Huchepie manor organic b&b is ideally located to visit the Loire Valley châteaux and the historical France heartland.
hautefort, France Bed & Breakfasts - 4 star French government rated bed and breakfast situated on beautiful perigord country estate--private cottages all over 250 years old including village chapel and village bakery
Arles, France Bike Tours - A charming tour from Avignon that visits the culturally rich countryside as far as the Mediterranean and back to Avignon - "Cite des Papes". This tour will find you traveling on small roads away from traffic and discovering the best of provence!
Argelès sur mer, France Language Schools - In our language school in the south of France,in Argelès sur mer, near the spanish border you'll find a wide variety of programs:intensive courses, one to one tuition,lessons duo and lessons by phone.We tailor make packages that suit you.
, France Bike Tours - Self-guided, guided. Many weekly and some departures. Alsace, Burgundy, Loire, Provence, Canal du Midi, Normandy, Pyrenees, from leisure to alpine challenges. 40 bike routes throughout France, 200 more in 30 countries across Europe.
Bordeaux, France Vacation Rentals - Bordeaux vineyards. 18th century "grange" in 2 acres of gardens with heated pool, 6 person spa. 4 bedrooms, WIFI. Ideal location for wine tasting, St Emilion, Bordeaux, Cognac etc. For slideshow...
, France River Cruises - We are your one-stop-shop travel experts, offering professional advice on canal and river cruising as well as vacation packages throughout France.
Sainte-Marie-sur-Ouche, France River Cruises - Experience the luxury of an All Inclusive Dream Cruise on board �La Reine Pedauque�", discover the magnificent French Burgundy canal and the beauty of the countryside. 4 or 7 day cruises, 4 suites for 8 passengers
Cannes, France Motorcycle Rentals - Motorcycle Rentals and Tours in Cannes, France. Hire a BMW, Ducati, Harley, KTM or MV Agusta and ride the Alps, Provence and French Riviera. Or join our Motorcycle Tours to enjoy the region's best roads with an English-speaking motorcycle guide.
Bayeux, France Cooking Schools - Create a classic Normandie dish in one night. Le Chef, Invites you to achieve your menu, with the delight of enjoying your own culinary success in a stylish personalised atmosphere. Diploma and Chef's hat awarded.
|Page: 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
France Vacations - Travel guide to France accommodations and vacation packages. Visit France for getaways and adventure travel. Plan your France vacation today.
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New Goldfinger Banana
bamboomersoonerMay 10, 2007
I just received a Goldfinger banana tree from Stokes Tropicals, arrived in great shape btw. So I planted in a spot where it will get plenty of sunshine and put plenty of peat humus around the root ball of the tree. At this point, do I need to fertilize? How often should I water? (Even though we have been getting a TON of rain lately)
Thanks for any input you all have.
cowboyup4christ(Zone 8)
I fertilize every two weeks and water at least once a day if it has not rained. bananas are ferocious feeders when they are growing and need lots of water. however win the winter when they are not growing do not water much at all as they will rot.


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We recommend
Product Info
Syncro 3/2 Back Zip GBS Wetsuit
100% Hyperstretch 3.0 neoprene Thermal Smoothie chest panel lined with BioFleece Thermal Lining Hydroshield water barrier lined with BioFleece Hydro wrap adjustable neck closureGBS (glued & blindstitched) seamsYKK standard length GBS back zipEcto flex knee pads
Enjoy Free Shipping on all orders to the US and Canada!
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REVIEW SNAPSHOT®by PowerReviews
Best Uses
- Was this a gift?:
- No (4)
Reviewed by 5 customers
Sort by
- cheap
- Cute
- Effective
- Warm At 45f
- Arm Length Long By 2cm
- Tight neck
- Torso Length Long By 5cm
- Velcro Pulls Out Hair
Best Uses
- Spring Surf
- Winter surf
Comments about roxy Syncro 3/2 Back Zip GBS Wetsuit:
My roxy suit is pretty great, but unfortunately I won't be buying Roxy suit anymore, o'neill has the best fit for me.
Not to discourage anyone. If you are taller than 5'4", weigh 110-125lbs, 34" bust, 25" waist, 35" hips, it's a perfect fit for a size 4 (unlike billabong's suits that have so much unnecessary fabric around the shoulders back and waist.) I wouldn't go smaller in size or else you'll feel like you're choking.
What I have to do now is alter my size 4 suit and take out the middle band between the color and the pant leg.
After a year of owning it, let's say for as much as I surfed, that would equate to once a week year round, the sharkskin panel is starting to unglue so slightly and some points of contact which I will fix with neoprene cement. The green and blue are starting to get dirty colored (no idea what to do with that).
With all the water running through the pant legs got me down, but I did champ out to 2 hour sessions with as low as 40F and I would wear this up to 50-55F water temperature.
I would buy Roxy more since I love the look and this alteration is going to by super easy.
- Was this a gift?:
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- Great Fit Hourglass Figur
- Kept me warm
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- Bodyboarding Surfing
Comments about roxy Syncro 3/2 Back Zip GBS Wetsuit:
This fit great even around my hips. I was able to stay in the water longer. It's cute I have the one with the blue accents. It does get cold in Hawaii with the wind in the winter. Probably like all wetsuits it was a little hard to get out of.
- Was this a gift?:
- No
- Flexible
- Good Fit
- Super warm
- Warm As A 43 Suit
Best Uses
- Surfing
Comments about roxy Syncro 3/2 Back Zip GBS Wetsuit:
Super warm, flexible & as warm as 4/3 full suit. Super stoked on this suit.
Surf just about everyday in it & has held up well. Great colors too lots of complements. Great price too, highly recommend it!
Surfed in 55 water & held well ;)
- Was this a gift?:
- No
- Comfort
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- Quality
- value
Best Uses
Comments about roxy Syncro 3/2 Back Zip GBS Wetsuit:
Love this wetsuit will be using to paddleboard in our lake. We live in the NE so this should let me get on my board a month earlier and stay on at least a month longer! Can not wait!!!
- Was this a gift?:
- Yes
- Kept Me Warm In 70' Water
- Looks and feels good
- Size 2 Perfect Fit
- The Air Temp Was 64'
- none
Best Uses
- I Use It For Body Surfing
- Surfing Too
Comments about roxy Syncro 3/2 Back Zip GBS Wetsuit:
I tried it out today when the water was 71 degrees and the air temperature was 64 degrees and windy. The suit fit perfectly and there was no scratching or itching. Without it I would not have spent as much time in the water bodysurfing, but I had an unexpected wonderful day because of this wetsuit. I got some nice compliment on how it looked.
- Was this a gift?:
- No


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Replicator resolution
View Single Post
August 5 2010, 08:52 AM
Re: Replicator resolution
We know Riker fishes.
We don't know if he eats what he catches, but that wouldn't be at odds with Riker's conviction that enslaving animals for food is wrong. He isn't enslaving anybody - he's hunting free animals in their natural environment. For all we know, he gets his eggs much the same way.
Joe Sisko also kills "real" animals for food. Or if his mollusks are artificially created, he's a sadist for creating them dirty and alive, and forcing his son to clean them, after which he himself can kill them for food. But again, "real" clams would not be in violation of the idea that animals must not be enslaved for food.
They can apparently be enslaved for other purposes, though, as several heroes keep pets...
Timo Saloniemi
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Water Monitoring Companies in Kazakhstan
Category: Monitoring & Testing ×
Subcategory: Water Monitoring ×
Industry: Manufacturing, Other ×
Location: Asia & Middle East ×
Location: Kazakhstan ×
Kurz Instruments, Inc.
based in Monterey, CALIFORNIA (USA)
Kurz Instruments has maintained a reputation for designing and manufacturing Thermal Mass Flow Transmitters for industrial gas flow measurement. Our engineers, product development specialists and management have developed flow metering products capable of operating in the harshest of environments. For more than 35 years our team has successfully provided solutions to our ...
Agape Water Solutions, Inc.
based in Harleysville, PENNSYLVANIA (USA)
Agape Water Solutions, Inc. provides industrial water treatment equipment and technical support services. We have unsurpassed knowledge and experience with membrane systems including reverse osmosis (RO), electrodeionization (EDI) and microfiltration (MF) / ultrafiltration (UF).
Inspectahire Instrument Company Ltd.
based in Aberdeen, UNITED KINGDOM
Inspectahire Instrument Company Ltd. provide inspection solutions and instruments for the environmental industry. We provide borescopes and cameras for ecologists to track wildlife. We inspect gas vents and buried pipework utilising explosion proof camera technology. Thermographic surveys are performed of buildings to demonstrate their heat insulation. Groundwater flow ...
CHEMetrics, Inc. (Water Analysis Test Kits)
based in Calverton, VIRGINIA (USA)
Chemetrics offers an extensive line of water testing kits. CHEMetrics supplies water analysis test kits for measuring more than 50 water analytes and parameters. Industries: water treatment, petroleum, chemical, power, metals, environmental, manufacturing, pulp/paper and food & beverage.
ThermoEnergy Corporation
based in Worcester, MASSACHUSETTS (USA)
ThermoEnergy is an integrated technologies company engaged in the worldwide development and sales of sustainable technologies for wastewater recovery and reuse. The company's water division is a leading provider of custom designed, turn-key systems for wastewater treatment and reuse, chemical/metals recovery, and water conservation. ThemoEnergy`s CAST® and RCAST ...
Pulsar Instruments plc
based in Filey, UNITED KINGDOM
Pulsar Instruments Plc have over 45 years of experience in the noise measurement sector. The company designs, manufactures and markets from their head office in the UK compliant, common sense, practical solutions for industrial, occupational and other noise measurement applications in the work environment via a global distribution network. From the beginning, Pulsar have ...
based in New Delhi, INDIA
ADVANCE INSPECTION & TESTING LAB (AITL) is a leading Delhi based test house having facilities and expertise comparable to national & international testing laboratories. The laboratory offers high quality & extremely cost effective Inspection & Testing services, to building and construction industries, industrial product manufacturer`s processors, Government and Public ...
Banner Engineering Corp.
based in Minneapolis, MINNESOTA (USA)
Banner is a global leader in process and factory automation. In addition to its traditionnal product lines, Banner Engineering provides innovative Wireless I/O solutions to the environmental industry helping gater data from virtually any sensor, detector or analyser in the field, and controlling equipment (pumps, valves, PLCs...) without costly wire runs.
Art's Manufacturing and Supply, Inc. (AMS Inc.)
based in American Falls, IDAHO (USA)
AMS, Inc. manufactures hand augers, soil probes, core samplers, sludge and sediment samplers, gas vapor probes. AMS PowerProbes can be mounted on pick-up trucks, tracks, or trailers, and have gained widespread acceptance due to their versatility, relatively low cost, and ease of mobility. They are designed to be used for: soil, soil gas, and groundwater sampling and ...
Modern Water - Monitoring Division
based in Cambridge, UNITED KINGDOM
Modern Water's Monitoring division has developed some of the world's most advanced systems for monitoring water quality. We provide market-leading toxicity monitoring systems, trace metal and environmental monitoring systems. The range includes systems for portable, online, and laboratory use. Our toxicity monitoring products, Microtox and Microtox CTM, detect toxins in ...


text:
- School
- Athletics
- Performing Arts
- MonMar30 B
- MonMar30 Quarter Three Ends
- TueMar31 T DAY
- ThuApr02 Half Day Session of School 12:12PM Dismissal
- ThuApr02 Holy Thursday
- ThuApr02 Lenten Spirituality Day(Morning Prayer) 9:30 AM to 10:30 AMSt. James Church
- SatMar28 Girls Softball - Varsity vs. Brick Memorial CANCELLED 9:00 AMWall Township
- SatMar28 Boys Baseball - Freshmen vs. North Brunswick CANCELLED (Away) 10:00 AM to 1:00 PMNorth Brunswick
- SatMar28 Boys Baseball - Junior Varsity vs. North Brunswick CANCELLED (Home) 10:00 AM to 1:00 PMCt. Basie Park
- MonMar30 Jazz Band Rehearsal 3:00 PM to 5:00 PMFAPA Building
- MonMar30 Senior Choreography Rehearsal 3:00 PM to 4:30 PMFAPA Building
- TueMar31 Choir Rehearsal 3:00 PM to 5:00 PMFAPA Building
RBC News
Red Bank Catholic has been ranked 23rd of the top overall high school Athletic programs in the country.
The RBC Make a Wish Club organized an event to raise funds for the NJ Chapter of the Make a Wish Foundation and while breaking a Guinness World record.
A group of students joined Mrs. Mary Jane Davis at Dorbrook Park for the 23rd Annual "Bluebird Day" to assist Cornell University's NestWatch Project.
RBC Performing Arts will be bringing the hilarious story of everyone's favorite ogre to life on stage. From March 12th through the 15th RBC will present "Shrek The Musical", the musical comedy based on the Oscar-winning Dreamworks film, "Shrek"!
RBC Ice Hockey wins back to back Shore Conference tournaments defeating St. John Vianney 8-0.
Red Bank Catholic High School112 Broad Street, Red Bank, NJ 07701
(732) 747-1774
Website generously provided by the Red Bank Catholic Parent Connect


text:
Executive Inn San Jose
No availability found through our partners. Please contact the business directly or check some of our recommended alternatives.
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TripAdvisor Reviews Executive Inn San Jose
Travel Blogs from San Jose
Reflections of Canada
... br>
That night we stayed in solitary splendour at Tete Jaune Cache, beside a small stream running into the mighty Fraser River. Earlier we'd visited Rearguard Falls, 800 miles up the Fraser, the furthermost migration for Pacific salmon. The powerful 6 metres high falls are the final hurdle before the few King Salmon who make it reach their ...
How it all came about
... ok.
We went to Sacramento to see Jon and Danica showing two of their Scottish Terriers before taking a look round the city.
Sacramento is the state capitol of California which, despite being 90 miles from the Pacific Ocean , is an important port - so the 'river' we walked along through the ...
Weekend Getaway in Sequoia Natl Forest, CA
... and a pretty drive to Lake Isabella during the day.
It is ~5 hours drive from the Bay Area and well worth a long weekend trip.
Remember - the closest gas station is miles away - maybe ~45-60 mins drive away so keep your fuel tank happy and full!! maybe carry an extra can if you have one!! Always better to be safe when on a fun trip....
Enjoy the pics and ...
... shoulder by Collie - grrrr!) and was served a hot chicken (I think that's what it was) dinner upon waking. The salad was a tad warm but I ate it nonetheless because that will be the last time I eat raw vegetables for the next two and a half weeks. After numerous trips to the bathroom and short walks up and down the aisles, I ran out of things to keep me entertained so I finally settled in my seat and watched four movies. I haven't watched that much television since the ...
San Fran San Jose
... 233;? We did, but first we drove by some of Ken's family's homes and Ken's old school. First we went to Foster City, then we went to Saratoga, where we drove past their old house and then Blue Hills Elementary school, and the kids got out and stretched their legs a bit by playing on the same playground area their dad did many years ago. Ken went to third grade there. Next we went into San José, where we swam, swam and swam! The weather was back to what ...


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Patterson on new leadership
by Ted Carter
Published: December 7,2012
Tags: Aubrey Patterson, BancorpSouth, banking, Tupelo
After 22 years as CEO of BancorpSouth, Audrey Patterson has handed the chief executive duties to veteran Texas banker James "Dan" Rollins III. Patterson, who will remain as chairman, recently answered several questions regarding the $13 billion banking regional banking company's new leadership.
>>New BancorpSouth CEO looks to repeat his Texas-style success in Tupelo
Major Challenges:
BancorpSouth like all commercial banks is facing a challenging economic environment with extremely low interest rates and limited lending opportunity. This will continue to put pressure on the earnings of all banks and under Dan's leadership BancorpSouth will have to continue working our way through these issues.
Dan Rollins' background:
Dan has been the president and chief operating officer of a high-performing regional bank. The basic foundation of both Prosperity and BancorpSouth is the strong network of community banks which comprise our systems. His leadership and prior success have direct application to the daily operations of BancorpSouth.
BancorpSouth already has a strong community bank presence in east Texas, one which has been very successful and has continued to grow. Without question, Dan brings demonstrated ability to build our Texas franchise to even higher levels of performance.
Dan has already proved that he doesn't need this piece of advice because he is already doing exactly what I would recommend, and that is to meet as many people throughout our bank as possible and spend the first month or two listening and learning the company. He is very much a hands-on, people-oriented leader and one whom I think will continuously build on his relationship with our team members.
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Franchise Index: Previous Season / Next Season
Record: 8-3-1, Finished 1st in NFL West Division
NFL Season Summary
Scored 260 points (21.7/g), 4th of 12 in the NFL.
Allowed 231 points (19.2/g), 4th.
Differential of 29 points (2.4/g), 4th.
Expected W-L: 6.8-5.2.
Takeaway/Giveaway Differential +18 (1.5/g), 1st.
Simple Rating System: Total -1.3, Offense: -1.2, Defense: -0.2, SoS: -3.8, versus averages of 0.0.
Coached by
Sid Gillman
Training Camp: University of Redlands (Redlands, California)
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The Lehigh Valley Conference has staged its final athletic events, with the league's 12 schools set to merge into the new Eastern Pennsylvania Conference this fall.
The LVC's swan song will go down as the Year of the Trojans.
The 2013-14 high school sports year featured a bit of everything in the Lehigh Valley, from controversial coaching decisions to individuals shining at the state level. The consistent excellence of Parkland's athletic programs stood out from the crowd.
The Trojans finished 2013-14 with 13 LVC championships, winning titles in sports ranging from football to tennis to lacrosse. They added LVC tournament titles in five sports — girls cross country, golf, boys tennis, girls tennis and boys track.
Parkland kept winning when its teams reached the District 11 playoffs. The Trojans captured 12 district titles, with the football, baseball and softball teams among those to win both LVC and district championships.
"You start with, no question, we've got kids," Parkland football coach Jim Morgans said this week. "We've got kids that are nurtured through the North and South Parkland programs. They're very competitive. I think our middle schools, the coaches there in all sports do a great job.
"But the No. 1 ingredient is, you've got to have kids. Our job is to make sure they stay competitive, stay interested and don't mess it up."
Parkland athletic director Jeff Geisel also pointed to the talent base as the top reason for the Trojans' success. Parkland is one of the largest high schools in District 11, with an enrollment of more than 3,200 students.
The Trojans also have quality facilities. But other schools have large student bodies and nice fields, too.
From Geisel's viewpoint, what separates Parkland is the cooperation across the board at the school. It starts with a supportive administration and school board. It extends down to the maintenance staff that makes sure the school's facilities are ready for events big and small.
"Things are clearly laid out with what you should and shouldn't do," Geisel said. "If something's needed, we find a way to provide it. You just don't find that elsewhere. People are happy to be here."
As much success as Parkland had in 2013-14, it didn't hold a monopoly on winning. Bethlehem Catholic, for instance, produced some of the top achievements in the area.
The Golden Hawk wrestling team won a fourth straight PIAA Class 2A dual-meet title and had three individual state champions. The softball team reached the PIAA 3A final, the football team won its first District 11 title since 2001, and the girls basketball program won LVC and District 11 3A championships.
Catasauqua's football team shattered scoreboards and school records while winning a Colonial League title and the District 11 2A championship. Saucon Valley's Elizabeth Chikotas and Palmerton's Austin Shupp were two of the top track athletes in Pennsylvania.
And who could forget the Central Catholic boys basketball team's historic start? The Vikings opened the season 29-0, something no boys basketball program in Lehigh Valley history had ever done.
All of those achievements deserve a spot in any Lehigh Valley high school sports yearbook for 2013-14. The cover, though, belongs to Parkland.
Attend enough athletic events in the Lehigh Valley and you're bound to find someone willing to express a degree of Parkland fatigue. People tire of seeing the Trojans win so often.
Those within Parkland's walls aren't ready to give up their title of the Lehigh Valley's top athletic program.
"It's a credit to our kids, the structure of the school, the administration, our basketball coach, our baseball coach, they all do an outstanding job," Morgans said. "The kids take it seriously. The kids want to work.
"I've never been any place else where it's like this," he added of all the winning. "That's incredible."
Twitter @mcall_smiller
Trojan title haul
Parkland's athletic program piled up championships in 2013-14. Here's a look at all the titles the Trojans won during the school year.
LVC championships
Girls Cross Country
Girls Volleyball
Girls Tennis
Boys Soccer
Girls Swimming
Boys Track
Boys Lacrosse
Girls Lacrosse
Boys Tennis
Boys Volleyball
LVC tournament championships
Girls Cross Country
Boys Tennis
Girls Tennis
Boys Track
District 11 championships
Girls Cross Country
Girls Tennis
Boys Soccer
Boys Tennis
Boys Basketball
Girls Basketball
Girls Swimming
Boys Volleyball
Girls Lacrosse
SoftballCopyright © 2015, The Morning Call


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The invention relates to a process of manufacturing D(-)-3-hydroxybutyric acid by breeding microorganisms capable of producing said acid in a nutrient medium containing certain specific carbon source; bacterial strains especially suitable in carrying out the process; an application of the process for obtaining such microorganisms; and uses of thus produced acid.
1. A process for producing D(-)-3-hydroxybutyric acid which comprises cultivating a mutant microorganism on an aqueous nutrient medium containing trace elements, an assimilable nitrogen source and a carbon source selected from the group consisting of carbon dioxide, glucose, fructose, saccharose, lactose, molasses, whey, methanol, ethanol, glycerin and spent lye from caprolactam synthesis under conditions and for a time sufficient to produce said D(-)-3-hydroxybutyric acid and isolating said D(-)-3-hydroxybutyric acid from the medium, wherein said mutant microorganism does not form 3-hydroxybutyric acid dehydrogenase and produces at least 100 mg/liter of D(-)-3-hydroxybutyric acid when cultivated on said nutrient medium for 30 hours at a temperature between 25� and 40� C. and at a pH value between 4 and 8.
2. A process according to claim 1 wherein the microorganism mutant used is the D(-)-3-hydroxy-butyric acid-producing mutant derived from Alcaligenes eutrophus ATCC 23440, Azotobacter chroococcum DSM 281, Bacillus megatherium ATCC 32, Zoogloea ramigera ATCC 19623, Clostridium butyricum ATCC 19398 and Mycoplana rubra CBS 385.76.
3. A process according to claim 1, wherein when the carbon source is selected from the group consisting of glucose, fructose, saccharose, lactose, molasses or whey, the concentration of the carbon source is 2-25% weight/volume, when the carbon source is selected from the group consisting of methanol, ethanol or glycerin the concentration of the carbon source is 1-10% weight/volume, and when the carbon source is spent lye from caprolactam synthesis the concentration of the carbon source is 1-10% weight/volume, calculated as the total quantity of carboxylic acids contained therein.
4. A process according to claim 1 wherein cultivation is effected under aerobic conditions with aerobes and under anaerobic conditions with anaerobes.
5. The process of claim 1, wherein said microorganism mutant is obtained by exposing a parent microorganism which yields said microorganism mutant to a mutagenic agent selected from the group consisting of ultraviolet light, nitro-nitrosomethyl guanidine, sodium nitrite and alkylating substances.
6. A process for producing a microorganism mutant capable of converting into D(-)-3-hydroxybutyric acid a maximum amount of a carbon source selected from the group consisting of carbon dioxide, glucose, fructose, saccharose, lactose, molasses, whey, methanol, ethanol, glycerin and spent lye from caprolactam synthesis within a minimum space of time, which comprises (1) subjecting a microorganism capable of excreting butyric acid or D(-)-3-hydroxybutyric acid, or of accumulating poly(D-3-hydroxybutyric acid), and which does not form the enzyme 3-hydroxybutyric acid dehydrogenase, to the action of a mutagenic agent, (2) selecting from the mutants thus obtained those which cannot metabolise D(-)-3-hydroxybutyric acid and (3) selecting from the thus obtained mutants those which are capable of producing at least 100 mg of D(-)-3-hydroxybutyric acid per liter of the nutrient medium within 30 hours when cultivated at a temperature of approximately 25�-40� C. and a pH value of approximately 4-8 in an aqueous nutrient medium which contains a carbon source as specified above.
7. The process of claim 6, wherein said mutagenic agent is selected from the group consisting of ultraviolet light, nitro-nitrosomethyl guanidine, sodium nitrite and alkylating substances.
The present invention relates to a microbiological method of manufacturing D(-)-3-hydroxybutyric acid, microorganisms for use in such a method and methods of obtaining such microorganisms.
In the present application optically active 3-hydroxybutyric acids are designated according to the following definitions. Naturally occurring 3-hydroxybutyric acid or D(-)-3-hydroxybutyric acid (also, D(-)-β-hydroxybutyric acid) has a specific rotation [α]D 25 of -24.5� at a concentration of 5 g per 100 ml of water while L(+)-3-hydroxybutyric acid which does not occur in nature has a specific rotation [α]D 10 of +24.3� at a concentration of 2.226 g per 100 ml of water.
The 3-hydroxybutyric acid concerned herein unless the context otherwise provides is exclusively the isomer which has a negative rotation on the sodium D line i.e. the naturally occurring isomer, which for the sake of simplicity is henceforth designated by the abbreviation β-HB.
Various methods have been disclosed for the manufacture of racemic DL-3-hydroxybutyric acid; see e.g. Deutsche Offenlegungsschrift 441 003 and Wislicenus, Ann. 149 (1869), 207. McKenzie and Harden have also succeeded in manufacturing optically active pure L(+)-3-hydroxybutyric acid from DL-3-hydroxybutyric acid using Aspergillus niger [J. Chem. Soc., 83 (1903), 430]. However neither of these two forms are generally present in nature.
D(-)-3-hydroxybutyric acid occurs in humans, animals, plants and microorganisms (i.e. bacteria, yeasts, fungus and algae) as an essential substance in fat metabolism [Edstrom et al., Acta Obstet. gynecol. Scand. 54 (1975), 347; Moore et al. Am,J. Physiol 230 (1976), 619]. Because of this natural occurrence and physiological function, the compound could deserve great significance in, among other things, parenteral nutrition and as a starting material in the manufacture of chemical compounds which are compatible with the human organism.
It is, however, surprisingly true that hitherto, in contrast to the racemic and dextrorotatory i.e. non-naturally occurring 3-hydroxybutyric acid, no technically feasible method has been described for the manufacture of D(-)-3-hydroxybutyric acid. In particular, no microorganism has previously been found capable of separating out the desired carboxylic acid.
Surprisingly we have now found that microorganisms can be obtained, which separate out high concentrations of D(-)-3-hydroxybutyric acid starting from extremely cheap carbon sources and which can be improved still further in this respect.
Microorganisms particularly suitable for starting with are those which are able to synthesise butyric acid or poly-(D-3-hydroxybutyric acid), such as e.g. members of the genera Azotobacteraceae, Bacillaceae, Pseudomonadaceae, Methanomonadaceae, Spirillaceae, Achromobacteraceae and Enterobacteraceae.
Thus, microorganisms known to accumulate poly-(D-3-hydroxybutyric acid), such as e.g. Alcaligenes eutrophus ATCC 23440, Azotobacter chroococcum DSM 281, Bacillus megatherium ATCC 32, Zoogloea ramigera ATCC 19623 and the methanol-utiliser Mycoplana rubra CBS 385.76 isolated from air, may be bred e.g. on fructose as the sole carbon source under submerged conditions in such a way that as little poly(D-3-hydroxybutyric acid) as possible is accumulated. Such conditions include small C/N ratios and/or high oxygen partial pressures in the nutrient medium. After removal of the carbon source the microorganisms are placed into a medium which contains racemic or D(-)-3-hydroxybutyric acid as the sole carbon source and an antibiotic which destroys active or growing cells. The surviving, resting or non-growing, cells which do not form the enzyme 3-hydroxybutyric acid dehydrogenase (=EC 18.104.22.168; see E. T. Barman, Enzyme Handbook, Springer-Verlag, Berlin 1969) and thus cannot metabolise β-HB, are isolated and tested for their capacity to form β-HB. For this purpose, the concentration of β-HB is determined quantitatively in aliquots of the culture medium containing the microorganisms according to the enzymatic method of Williamson & Mellanby (Methods of Enzymatic Analysis, Verlag Chemie GmbH, Weinheim and Academic Press, Inc., New York 1974, Page 1883). The microorganisms capable of synthesising and separating out β-HB are selected and treated with mutagenic agents, e.g. with ultra-violet light, NaNO2 or nitro-nitrosomethyl guanidine (see e.g. R. C. Clowes and W. Hayes, Experiments in Microbial Genetics, Blackwell Scientific Publications, Oxford and Edinburgh 1968). From the mutants thus produced may be selected those which produce, per unit volume of the culture medium, even higher concentrations of β-HB than the original strain.
In order to still further increase the capacity to produce D(-)-3-hydroxybutyric acid in the strains thus selected, they can be further mutated and of the resultant mutants, those may desirably be selected which reveal not only missing or impaired D(-)-3-hydroxybutyric acid dehydrogenase activity, but also missing or impaired activity of one or more enzymes of the tricarboxylic acid cycle (see Barman, loc. cit.).
On the other hand, microorganisms which do not accumulate poly-(D-3-hydroxybutyric acid) but form butyric acid, such as, for example, the Clostridium types, e.g. Clostridium butyricum ATCC 19398 may also be treated in a similar way, i.e. those organisms are selected which do not form any 3-hydroxybutyric acid dehydrogenase and, as a result, cannot metabolise β-HB. Thus, such strains may be bred on agar plates as scattered colonies. After growth has taken place, the agar surfaces may be spread with a mixture of 3-hydroxybutyric acid dehydrogenase, nicotine-adenine-dinucleotide in oxidised form (NAD) and a redox indicator consisting of methylene blue or triphenyltetrazolum chloride. A search may then be made for colonies which cause colour change of the redox indicator. These may be isolated and tested for their capacity to produce β-HB. Strains producing β-HB may be subjected to the mutation process mentioned above, in order to obtain derivatives with increased β-HB productivity.
According to the present invention, therefore, microorganisms may be selected which convert the maximum proportion of the carbon source into D(-)-3-hydroxybutyric acid in as short a time as possible by a method which comprises subjecting a microorganism capable of separating out butyric acid or D(-)-3-hydroxybutyric acid or of accumulating poly-(D-3-hydroxybutyric acid) to the action of mutagenic agents; selecting from the mutants thus obtained those which cannot metabolise D(-)-3-hydroxybutyric acid; and further selecting from the thus-selected mutants those which, optionally after further treatment with mutagenic agents, produce more D(-)-3-hydroxybutyric acid than the original microorganism when grown on a specific carbon source and under conditions mentioned below.
The microorganisms producing β-HB are advantageously accustomed by known methods to grow in high concentrations of specific carbon sources, i.e. carbon dioxide, assimilable carbohydrates, including e.g. glucose, fructose, saccharose, lactose, molasses and whey, assimilable alcohols, including e.g. methanol, ethanol and glycerin, and spent lyes from caprolactam synthesis, and thereby to produce β-HB. There can be added as a nitrogen source ammonium ions, and the usual trace elements as well. The derivatives of Alcaligenes eutrophus ATCC 23440 can, in addition to organic carbon sources such as those mentioned above, also metabolise carbon dioxide, if the atmosphere simultaneously contains molecular hydrogen and oxygen, in order to produce β-HB.
Preferred selected microorganisms thus obtained include mutants separating out D(-)-3-hydroxybutyric acid derived from Alcaligenes eutrophus ATCC 23440, e.g. the mutant CBS 381.76; from Azotobacter chroococcum DSM 281, e.g. the mutant CBS 383.76; from Bacillus megatherium ATCC 32, e.g. the mutant CBS 382.76; from Zoogloea ramigera ATCC 19623, e.g. the mutant CBS 384.76; from Clostridium butyricum ATCC 19398, e.g. the mutant CBS 380.76; and from Mycoplana rubra CBS 385.76 and mutants thereof. The above-mentioned strains are filed at the American Type Culture Collection (ATCC) 12301 Parklawn Drive, Rockville (Maryland, U.S.A.): Centraal Bureau voor Schimmelcultures (CBS), Oosterstraat 1, Baarn (Netherlands); and Deutsche Sammlung fur Mikroorganismen (DSM), Munich (BRD).
The microorganisms mentioned above, of course, can still further be improved in respect of their productivity by applying thereon the beforesaid methods.
To ensure that high yields are obtained in the large-scale manufacture of D(-)-3-hydroxybutyric acid, microorganisms are suitable which can produce at least 100 mg of the acid per liter of nutrient medium when bred under the conditions specified below and for a breeding period of 30 hours. It is, however, more preferred to use microorganisms whose production of β-HB is at least 200 mg/liter of nutrient medium for 30 hours under the same breeding conditions. Particularly preferred are microorganisms with a β-HB production (for 30 hours) of at least 500 mg/liter or even at least 1 g/liter of nutrient solution.
The carbon source content in the nutrient solution is advantageously approximately 2-25% glucose, fructose, saccharose, lactose, molasses or whey, calculated as pure carbohydrate, approximately 1-10% methanol, ethanol or glycerin or approximately 1-10% spent lye from caprolactam synthesis, calculated as the total amount of carboxylic acid contained therein.
Although all the microorganisms obtained according to the method of the present invention are capable of producing β-HB on solid nutrient media, liquid media are preferred for practical reasons. The microorganisms may be cultured according to conventional techniques as is well known in the art. Thus aqueous nutrient solutions having the carbon source concentration mentioned above and with an assimilable nitrogen source and the necessary trace elements may be prepared in known manner. The sterile nutrient media may, for example, be innoculated with 0.1-10% by volume of a preliminary culture and then incubated for 8-72 hours at approximately 25�-40� C. either in stationary culture or with exclusion of air for anaerobic microorganisms, e.g. the Clostridium types, or with stirring or shaking and optionally with a supply of air or a mixture of carbon dioxide, hydrogen and oxygen (or air) for aerobic microorganisms. To obtain as high a yield as possible of the desired D(-)-3-hydroxybutyric acid, the pH value of the inocculated solution is preferably maintained at 4-8 by the addition of a sterile alkali solution. During the growth of the culture, additional carbon source material may be added continuously, if desired, in order to maintain a constant concentration and achieve maximum production of β-HB.
Hence, the process according to the invention comprises breeding a microorganism, which produces D(-)-3-hydroxybutyric acid, at a temperature of approximately 25�-40� C. and a pH value of 4-8 in an aqueous nutrient medium which contains as carbon source carbon dioxide, an assimilable carbohydrate from the group glucose, fructose, saccharose, lactose, molasses and whey, an assimilable alcohol from the group methanol, ethanol and glycerin or the spent lye from caprolactam synthesis, together with an assimilable nitrogen source and trace elements, and isolating the D(-)-3-hydroxybutyric acid formed from the culture liquor by known methods.
After completion of fermentation, i.e. when the maximum concentration of β-HB is reached, the β-HB may, for example, be extracted by means of ion exchange resins optionally after separation of the culture mass from the fermented solution or, after acidification of the fermented solution to approximately pH 1.5-2.5, by means of solvents not miscible with water, such as e.g. ether, chloroform or other chlorinated hydrocarbons, ethyl acetate or other esters, isopropanol, butanol and other alcohols. β-HB can be precipitated from the organic solutions as a metal salt, e.g. the lithium, potassium, sodium, calcium, barium, nickel or zinc salt, or may be transformed into a concentrated aqueous solution.
D(-)-3-hydroxybutyric acid can be used especially in parenteral nutrition as it can replace in part or in full the fatty constituents used hitherto with regard to calorie supply, since it is substantially equivalent to the fats in this respect. In addition, however, it has the advantage that it is immediately available for intermediate metabolism, as compared with fats, which require either initial saponification and repeated β-oxidation.
As is also known odoriferous and aromatic substances occurring in fruit and plants consist partly of lower alkyl esters of D(-)-3-hydroxybutyric acid. Their synthetic manufacture, has, however, suffered hitherto from the impossibility of manufacturing D(-)-3-hydroxybutyric acid in a pure state by a technically feasible and economically productive method. In particular, no microorganisms have been known hitherto which separate out the desired carboxylic acid.
The method according to the invention, however, enables substantially pure D(-)-3-hydroxbutyric acid to be manufactured with a high yield from extremely cheap carbon sources and thus leads to the possibility of synthesising such optically active esters.
Thus the D(-)-3-hydroxybutyric acid manufactured according to the invention may be esterified with an optionally substituted lower alkanol according to known methods. For example, esterification may, for example, be carried out by treatment of the carboxylic acid with an excess of alkanol, particularly a C1-5 alkanol, in the presence of a strong mineral acid, e.g. sulphuric acid.
The esters thus obtained have the same configuration as the corresponding odoriferous and aromatic substances occuring in fruit and plants. In contrast with the aromatic fruit substances manufactured hitherto using racemic DL-3-hydroxybutyric acid, the esters of D(-)-3-hydroxybutyric acid have the "natural" configuration which means that they are toxicologically harmless and, on a weight basis, have a significantly increased olfactory and organoleptic activity. Esters manufactured in this way may be used as odoriferous substances and aromatic substances in, for example, foodstuffs and perfumery.
The following examples illustrate the invention.
A nutrient solution is first prepared which contains 0.9% Na2 HPO4.12 H2 O, 0.15% KH2 PO4, 0.1% NH4 Cl, 0.04% MgSO4.7 H2 0, 0.0195% iron (II)-NH4 -citrate .6 H2 O, 0.00133% CaCl2 and, per liter of solution, 1 ml of a trace element solution containing 0.01% ZnSO4.7 H2 O, 0.003% MnCl2.4 H2 O, 0.03% boric acid, 0.02% CoCl2.6 H2 O, 0.001% CuCl2.2 H2 O, 0.002% NiCl2.6 H2 O and 0.003% Na2 MoO4.2 H2 O. In 250 ml Erlenmeyer flasks, shaken or stationary cultures are bred, each with 30 ml of nutrient solution, at 30� C. for 16-24 hours. 20 ml of sterile-filtered fructose solution containing 10 g of fructose are added per 980 ml to the solution sterilised at 121� C. for 20 minutes. Inoculation is carried out by means of a loop from a 24-hour slant agar culture.
The shaken culture is sedimented on a centrifuge and the culture mass is washed with water and then suspended in approximately 4 ml of physiological saline solution. After approximately 1 hour 4 ml 0.6 N sodium acetate buffer of pH 4.6 and 4 ml of a 0.05 N sodium nitrite solution are optionally added to produce mutation and the culture is incubated for a few minutes. After washing, the culture mass of each microorganism (see following table) is distributed between 10 250 ml Erlenmeyer flasks to which are added the above medium except that it contains 2% DL-3-hydroxybutyric acid (or 1% D(-)-3-hydroxybutyric acid) instead of the fructose. After clouding has visibly started to increase in the flasks, each flask has added to it, depending on the organism, 1 ml of a sterile-filtered solution of 2-1000 μg/K-Penicillin G, 1 ml of a sterile-filtered solution of 30-1000 μg/ml Bacitracin, 1 ml of a sterile-filtered solution of 10-2000 μg phosphonomycin/ml, 5-1000 μg vancomycin/ml or mixtures of these and/or other antibiotics, such as e.g. colistin sulphate, which influence formation of the cellular wall or division of the microbes. After varying incubation periods, i.e. after 1-16 hours, the antibiotics are washed out or destroyed with penicillinase. After further washing of the culture mass the surviving microorganisms are inoculated into flasks containg a fructose medium and are incubated.
After 16-36 hours a series of dilutions are made from the cultures in each flask and from the dilutions, containing approximately 50-1000 organisms per ml, 0.1 ml are smeared on agar plates to obtain individual scattered colonies. The agar contains, as the sole carbon source, 2% sodium DL-3-hydroxybutyrate and 0.008% fructose. After approximately 48 hours a large number of small colonies of varying appearance of approximately 1/4-1 mm diameter and colonies of normal size have formed. From each of the small colonies inocula are transferred by means of sterile dental slivers of wood, some onto a spot on an agar plate where sodium (DL)-3-hydroxybutyrate is the sole carbon source and others onto spots on an agar plate where fructose or glycerin is the sole carbon source. The desired strains, which can no longer metabolise β-HB and which possibly produce β-HB are to be found among those spontaneously present or artificially produced mutant colonies which cannot grow or can grow only poorly on 3-hydroxybutyric acid due to the absence or impairment of D(-)- 3-hydroxybutyric acid dehydrogenase, but thrive well on an assimilable carbohydrate such as e.g. fructose or glycerin. The absence or impairment of D(-)-3-hydroxybutyric acid dehydrogenase activity is a prerequisite for a practically useable method of producing D(-)-3-hydroxybutyric acid.
These strains are over-inoculated on milky-cloudy lime agar prepared as follows: a solidified agar layer approximately 5 mm thick containing the nutrient solution mentioned in the introduction, provided with a suitable carbon source, is covered with a layer of the same sterile nutrient solution, which contains additionally 0.1% of pure, precipitated CaCO3 in such a way that an upper layer approximately 1-2 mm thick arises. Colonies whose peripheral agar area lost its milky cloudiness and which were therefore surrounded by a transparent areola were found to produce acids.
Shaken cultures were made of these colonies in suitable submerged media in order to test their capacity to produce β-HB by the enzyme method of Williamson & Mellanby (loc. cit., see above). Good acid producers were inoculated by means of a loop from a slant agar culture into 500 ml Erlenmeyer flasks each containing 100 ml of nutrient solution (see above), each with 5 g/l of the appropriate carbon source (see Table) and the flasks were then incubated on a rotary shaker at 135 rpm and 30� C. After 30-52 hours samples were taken and analysed. The results are set out in following Table.
__________________________________________________________________________Production of β-HB-producing microorganisms10,000 strains per type were analysed for acid-forming capacity Strains with Number of βHB producing maximum β-HB Antibiotic strains productionMicro- treatment up to up to up to in Designation of Carbonorganisms with 1 mg/L 10 mg/L 95 mg/L mg/L best strain Source__________________________________________________________________________AlcaligeneseutrophusATCC 23440 -- 0* -- -- -- --and mutants Penicillin/ GA-3thereof Bacitracin ca.800 6 1 54 (CBS 381.76) FructoseBacillusmegatheriumATCC 32 -- 0* -- -- -- --and mutants Penicillin ca.600 15 2 65 GB-1 Saccha-thereof (CBS 382.76) roseAzotobacterchroococcumDSM 281 -- 0* -- -- -- --and mutants GC-7 Saccha-thereof Penicillin ca.700 7 3 39 (CBS 383.76) roseZoogloearamigera -- 0* -- -- -- --ATCC 19623and mutants Phosphono- GZ-1 Saccha-thereof mycin ca.100 2 0 6 (CBS 384.76) roseClostridiumbutyricumATCC 19398 -- 0* -- -- -- --and mutants Bacitracin/ GCl-2 Saccharosethereof Penicillin ca.1'100 3 1 76 (CBS 380.76)Mycoplanarubra -- 0* -- -- --and mutants D-Cycloser- GM-1 Glycerinthereof ine ca.300 21 0 8 (CBS 385.76) or Methanol__________________________________________________________________________ *Detectable neither in plate test nor enzymatically.
In a similar way, it was possible to obtain mutants which separate out β-HB in isolatable quantities by the application of other mutagenic agents, such as e.g. alkylating substances or radiation with e.g. ultra-violet light, or by isolating random spontaneous mutants.
In tests using Clostridium butyricum, the procedure is carried out in a similar way but under strict anaerobic conditions according to A. J. Schocher ["A Contribution to the Knowledge of Growth and Fermentation Physiology in Saccharolytic Clostrides", Dissertation, Laboratory for Microbiology, technische Hochschule Delft (Netherlands), 1959].
The β-HB producers GA-3 and GB-1 are subjected to the mutant selection process described in Example 1 and smeared onto agar plates in a similar way. After growth has finished, the colonies are applied by means of velvet stamps onto lime agar plates or fresh agar plates containing quantities of sterile bromocresol green or bromocresol purple as a pH indicator such that the agar is coloured either blue or purple. After short incubation periods, i.e. after only 5-6 hours, colour change of the pH indicator can be established, as a result of which colonies may be located which grow quickly and form a lot of acid. By means of this technique such colonies are selected and bred under submerged conditions in order to find their β-HB productivity. Most mutants of the β-HB producers used produce less β-HB or sometimes even no β-HB at all. However, among the many mutants obtained by mutation a few are perceived which produce more β-HB than the original strain. Such mutants are subjected to a new mutation/selection cycle in order to detect the strains which can produce even higher concentrations of β-HB.
The selected strains were bred at 30� C. in shaken culture tubes (test tubes 16 mm in diameter) containing 5 ml of sterile nutrient medium, which are fastened at an angle of 45� to a plate oscillating horizontally at 135 rpm. In this way it was possible to obtain from the strain Alcaligenes eutrophus GA-3 (CBS 381.76) a culture which produced 95 mg β-HB/L on 5% fructose in 51 hours. A new mutation/selection process supplied a culture which produced approximately 400 mg β-HB/l under the same conditions.
It was possible to derive in a similar way from the strain Bacillus megatherium GB-1 (CBS 382.76) a shaken culture which formed 145 mg β-HB/L on saccharose within 54 hours. A new mutation/selection process supplied a shaken culture which produced more than 1 g β-HB/L under the same conditions.
The yields of β-HB could be increased still further by additional mutation/selection processes.
The mutant 31 N selected from Bacillus megatherium strain GB-1 was incubated in a 5-liter small fermenter (brand Biostat V, Braun-Melsungen) with 2.5 liters of the nutrient solution of Example 1, which contained, instead of fructose, 24 g of saccharose per liter and 100 mg peptone per liter. 100 ml of a culture in the same medium shaken overnight served as an inoculum. Air was blown through the small fermenter at 30� C. with approximately 0.5 volume/volume minute and agitated at 600 rpm. During breeding the pH was kept between 7.2 and 7.3 by the addition of 10% sodium hydroxide solution. After 30 hours the fermented solution contained 170 mg β-HB/liter. The product was absorbed from the supernatant fluid on Amberlite 410A in OH form and was eluted with 0.5 N sulphuric acid. The highly concentrated eluate was treated by the extraction process of P. A. Schaffer and W. M. Mariott [J. Biol. Chem., 16, 268 (1913)], as a result of which it was possible to isolate 330 mg of the calcium-zinc mixed salt of β-HB. After recrystallisation 3 times from ethanol/water the result was a product whose solution (8.027 g per 100 ml H2 O) at a layer thickness of 20 cm gave a rotation of the Na D line of -2.60�, which corresponds to a [α]D 20 of -16.2 agreeing with the literature.
In a similar way to Example 3, a medium was used which contained glucose instead of saccharose. It was possible to isolate 350 mg calcium-zinc D(-)-3-hydroxybutyrate.
In a similar way to Example 3, a medium was used which contained, instead of saccharose, 50 g of beet molasses per liter with a saccharose content of 51%. The resultant fermented solution contained 160 mg β-HB/liter after an incubation of 31 hours.
In a similar way to that above, saccharose was used with the same success.
Under anaerobic conditions the mutant GCl-112 from Clostridium butyricum ATCC 19398 was incubated for 48 hours in a medium containing 4% saccharose, 0.3% NH4 acetate, 0.178% Na2 HPO4.2H2 0, 0.136% KH2 PO4, 0.042% MgSO4.7 H2 O, 0.00025% FeSO4.7 H2 O, 0.0013% MnCl2.4 H2 O and 1 μg (+)-Biotin per 1000 ml, after which the fermented medium had a concentration of 171 mg β-HB/liter.
To find microorganisms capable of utilising methanol, a 500 ml beaker was filled with approximately 100 ml of the nutrient solution of Example 1 except that it contained 1% methanol as the sole carbon source. After a few days the culture was cloudy and individual scattered colonies could be produced from it after appropriate dilutions with water on agar plates with the medium from Example 1. From the selected colonies, shaken cultures are bred in the glycerin medium mentioned above and are analysed according to Law and Slepecky [J. Bact., 82 (1961), 33] for poly-β-hydroxybutyric acid production. In this way, a rod-shaped microorganism is obtained which forms up to 50% by weight of its cellular mass of poly-β-hydroxybutyric acid and has been identified by the curator of the National Collection of Industrial Bacteria (Ministry of Agriculture, Fisheries and Food, Torry Research Station, Aberdeen, Scotland) as a Mycoplana rubra strain. The strain has been registered at the Centraal Bureau voor Schimmelcultures under the designation CBS 385.76 and named originally Bacterium in editio Pl. It was characterised as follows:
Morphology (GP Agar, consisting of 1% glycerin+1% peptone at pH 7.0 on agar, 6 days at 30� C.). Pleomorphous rods 0.8�1.5-5.0 μm, cells contain a high percentage of poly-β-hydroxybutyric acid.
Older submerged cultures, i.e. more than 7 days old, contain small quantities of cells with primary cellular branches which are detectable under a phase-contrast microscope.
Colony (GP Agar, 6 days at 30� C.).
Pink-coloured pigment, transparent, round, edged, convex, smooth and shining, diameter 0.5-1.0 mm. Pigmentation is intensified with increasing age. Very poor growth on blood agar base (OXOID CM 55).
Submerged growth (GP nutrient solution, consisting of 1% glycerin+1% peptone at pH 7.0, 6 days at 30� C.). rather weak growth, forms surface ring and membrane, slightly viscous sediment.
At 10� C., little or no growth
At 15� C., growth
At 30� C., growth
At 37� C., no growth
Gram Staining negative to variable
Mobility +
Kovacs' Oxidase +
Catalase +
Hugh and Leifson (Glucose) OF-
Peptone water carbohydrates: acid formation on glycerin. no acid formation on glucose, fructose, maltose, saccharose, lactose, starch or mannitol.
Indol production -
Methyl red -
Voges-Proskauer -
Koser's Citrate -
Tryptone Weak formation of NH4 + -
Nitrate reduction -
Urease (Christensen) +
Starch Weak hydrolysis
Gelatine Not hydrolysed
Casein Not hydrolysed
Haemolysis -
Resistant to penicillin, chloramphenicol and polymyxin B; sensitive to streptomycin and tetracyclin.
Can utilise methanol but not methane as sole carbon source.
A procedure similar to that of Example 3 was carried out with the derivative GM-37 of the Mycoplana rubra strain (CBS 385.76) freshly isolated from air and utilising methanol, except that in the medium the saccharose was replaced by 2.5% methanol. After an incubation period of 48 hours at 30� C. the fermented solution contained 36 mg β-HB/liter.
The mutant H-7 selected from A. eutrophus strain GA-3 (CBS 381.76) was bred in a similar way to Example 3 in a small fermenter in the medium of Example 1, but without an organic carbon source and with aeration with 0.11 volume/volume.minute of a gas mixture containing 10% CO2, 20% O2 and 70% H2 at 28� C. with agitation, the pH value being kept at 7.0 by the addition of 10% NaOH. After 51 hours the fermented solution contained 30 mg β-HB/liter.
A further method which was used successfully to detect and select strains with high β-HB productivity comprised analysing cells obtained according to Example 1 and 2 in a short experiment for their β-HB formation kinetics. For this purpose, the cellular mass of a shaken culture 30 hours old was centrifuged, washed with a physiological common salt solution and suspended in fresh medium with a suitable carbon source and incubated e.g. in a Warburg respiratory apparatus. Cellular suspensions with higher capacity to form β-HB were distinguished by high O2 absorption. The β-HB formation kinetics of such suspensions were determined by enzymatic analysis. Strains with substantially increased β-HB formation capacity in relation to the parent strain were plated out in order to obtain pure clones for the production of β-HB.
In this way, it was possible to find from B. megatherium GB-1 sub-strains which secreted e.g. on molasses per minute and gramm of cellular dry mass 1.5 mg β-HB (e.g. the mutant 24 N). In the same test set-up it was possible to establish, for example, that the strain B. megatherium reaches its maximum production rate of β-HB at pH 5.
Further observation showed that mutants whose synthesis capacity in the citric acid cycle is disturbed form considerably higher quantities of β-HB than their normal parent strains. For example, the B. megatherium strain 24 N was treated with the deadening method using penicillin described in Example 1, having been previously treated with a mutagenic agent. Those mutants were selected and enriched which could not grow or could only grow poorly without additions of citric acid, succinic acid and glutamic acid. In this way, 4 mutant colonies were found which did not grow to an optimum degree without the addition to the medium of small quantities of the above-mentioned organic acids. One of these 4 mutants produced in a shaken culture using the medium of Example 1, with 1% saccharose and additional 0.05% meat extract, 0.01% citric acid, 0.01% succinic acid and 0.01% glutamic acid, in a 15-hour incubation, 11.75 times the quantity of β-HB that its parent strain 24 N produced under the same conditions.
Two groups of six young rabbits, each of approximately 500 g average body weight, were fed exclusively by means of a Vena cava catheter (Vena jugularis julema) at an infusion rate of 6 ml/hour. The animals of the control group received a solution containing 695 kcal per liter, i.e. approximately 17 g of peptides plus amino acids (=11 cal %), approximately 97 g carbohydrates (=48 cal %) and approximately 34 g fat (=41 cal %) and mineral substances and vitamins. In the test group 20% by weight of the fats were replaced by sodium D(-)-3-hydroxybutyrate. After 10 days the average increase in weight of the animals of both groups was identical. An increase in the proportion of D(-)-3-hydroxybutyric acid to 30% by weight of the fat constituents of the infusion solution led after 5 days to the same increase in weight as in the control group.
1 g of the D(-)-3-hydroxybutyric acid is dissolved in 100 ml of ethanol and approximately 1 ml of concentrated H2 SO4 is added to the resultant solution. The reaction mixture is stirred for 3 hours at room temperature with a magnetic stirrer and then diluted with 1 volume of water and treated with a quantity of diethyl ether so as to result in a 2-phase mixture. The organic phase is thoroughly washed with water and the ether is then distilled off under vacuum. The residue contains ethyl D(-)-3-hydroxybutyrate with a raspberry-like aroma; boiling point 184�-185� C./755 mm, [α]D 25 =-10.5� (not completely pure optically). On a weight basis the ester has considerably stronger aromatic properties than the ethyl ester of DL-3-hydroxybutyric acid obtainable commercially.
The extract of the fermented solution of Example 2, using Bacillus megatherium ATCC 32 strain CBS 382.76, is treated with an equal volume of ethanol, approximately 25 g Na2 SO4 and 3 ml concentrated H2 SO4 and stirred for 5 hours. The reaction mixture is then washed with a lot of water and the organic phase is then concentrated under vacuum. The same ethyl ester is obtained with the same properties as described in Example 13.
The procedure is conducted in a similar way to that of Example 14, but methanol is used instead of ethanol. From the evaporation residue methyl D(-)-3-hydroxybutyrate is obtained which has the aroma of fruit, boiling point 67�-68.5� C. [α]D 20 =-20.9� (without solvent).
The procedure is carried out in a similar way to that of Example 14 using the fermented solution of Example 6 instead of Example 2 and propyl alcohol instead of ethanol. The result is similar to that of Example 15.
The procedure is carried out in a similar way to that of Example 14 using the fermented solution of Example 7 instead of Example 2 and butyl alcohol instead of ethanol. The result is similar to that of Example 15.
The procedure is conducted in a similar way to that of Example 14 using the fermented solution of Example 9 instead of Example 2 and amyl alcohol instead of ethanol. The result is similar to that of Example 15.
Patent Citations
Non-Patent Citations
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Giovanni Francesco Barbieri known as Guercino (Cento 1591-Bologna 1666)
The Libyan Sibyl 1651
Oil on canvas | 116.3 x 96.5 cm (support, canvas/panel/str external) | RCIN 405340
Picture Gallery, Buckingham Palace
Giovanni Francesco Barbieri known as Guercino (Cento 1591-Bologna 1666)
The Libyan Sibyl 1651
Royal Collection Trust/© Her Majesty Queen Elizabeth II 2014
This late work by Guercino ('The Squinter') is one of a pair of Sibyls painted for Ippolito Cattani of Bologna. It depicts the Libyan Sibyl who prophesied the coming of Christ to the Gentiles. She is usually shown holding a lighted torch, but Guercino identifies her here by the inscription on the book.
This painting depicts one of the twelve Sibyls, who were the seers of classical antiquity alleged to have foretold the coming of Christ, and were thus adopted by the Church as pagan equivalents of the Old Testament prophets. Michelangelo alternated prophets and Sibyls on the ceiling of the Sistine Chapel, a prototype which may explain the great popularity of the subject over the next two hundred years. Guercino painted Sibyls throughout his career, especially in the 1640s and 1650s. The Libyan Sibyl had prophesied the manifestation of Christ to the gentiles. Guercino has not included her usual attribute of a lighted torch here, identifying her instead by means of the inscription on the book.
This is an example of Guercino's late work, in which the strong chiaroscuro of his early years has been lightened with paler tonalities and his style has become refined and classical. The effects of light and dark, with the Sibyl's face caught in half-shadow, recall his earlier work, but are more subtle, the handling of paint more delicate and economical. The sensuality of the flesh and the warmth of the drapery give the figure more humanity than comparable figures by Guido Reni: the cool blue of her dress is set off by the striking clash of colours in the pastel pink and orange brown of her mantle and the deep red, possibly velvet, of her dress. The few colours are subtly modulated and simply counterbalanced by the white of her turban and the knotted sash at her waist. The Sibyl has a noble grandeur despite the modest size of the painting. Her concentration on her reading, oblivious of the viewer, gives a poetic quality to the painting. The Sibyl's dress was originally higher across her left shoulder, but there appear to be few other visible pentimenti.
The painting has been identified as one of two half-length Sibyls painted for Ippolito Cattani (or Cattanio) of Bologna, for which Guercino received 120 ducatoni (or 150 scudi) on 4 December 1651. The pendant is The Samian Sibyl (private collection): the pairing was apt, since both Sibyls were associated with the island of Samos. A workshop replica of The Samian Sibyl was sold to the Grand Duke of Tuscany in 1777 and appears in the foreground of Johan Zoffany's Tribuna also in the Royal Collection of c.1772-8, possibly as a compliment to King George III on his excellent purchase of the present work. Whereas the Libyan Sibyl remains absorbed in her reading, the Samian Sibyl looks heavenwards, her hands resting on her book, her turban a rich green and red. Both works have the distinctive spelling of the word 'sibyl' as 'sybilla', which Guercino seems to have corrected a few years later to 'sibylla'.
Catalogue entry adapted from The Art of Italy in the Royal Collection: Renaissance and Baroque, London, 2007
ProvenanceIppolito Cattani, 1651; probably bought for George III by Richard Dalton in Italy, early 1760s
People involved
Giovanni Francesco Barbieri known as Guercino (Cento 1591-Bologna 1666) (artist)
Physical properties
Medium & Techniques
Oil on canvas
116.3 x 96.5 cm (support, canvas/panel/str external)
147.0 x 127.1 x 10.5 cm (frame, external)
Object categorie(s)
Object type(s)
ML : Levey, M., 1964. The Later Italian Pictures in the Collection of Her Majesty The Queen, Cambridge - ML 521
XQG 2004 GIII : Exhibited at the Queen's Gallery, London, George III & Queen Charlotte. Patronage, Collecting and Court Taste (2004) - XQG 2004 GIII 154
XQG 1988 : Exhibited at the Queen's Gallery, London, Treasures from the Royal Collection (1988) - XQG 1988 Treas 27
XQG 1977 Jubilee : Exhibited at the Queen's Gallery, London, Silver Jubilee Exhibition: The Queen's Pictures. The Story of the Royal Collection from Henry VIII to Elizabeth II (1977) - XQG 1977 Jubilee 31
XQG 2007 Italian : Whitaker, L., Clayton, M., 2007. The Art of Italy in the Royal Collection - Renaissance & Baroque, London - XQG 2007 Italian 108


text:
WorldCom Files Suit Against Former Controller, Wants Him to Repay $800,000
WorldCom, the wounded telecommunications giant, wants its former controller to repay nearly $800,000 the company gave him two years ago to retain his services.
WorldCom, the wounded telecommunications giant, wants its former controller to repay nearly $800,000 the company gave him two years ago to retain his services.
GE Access expects a very strong second half of the year, indicating that an economic recovery is well under way, executives said at the distributor's annual New Frontiers Conference here.
Intel Corp., which has avoided mass layoffs during the technology downturn, said Tuesday it is cutting 4,000 jobs as the chip-making giant posted lower-than-expected second-quarter earnings.
A group of computer and telecommunications companies led by Intel is in talks about forming a company that would proliferate Wi-Fi hotspots across the country, according to a report in the New York Times Tuesday.
SonicWall Tuesday took the wraps off a new firewall/VPN appliance with automated fail-over technology that supports both broadband and dial-up connections.
Computer Associates International Tuesday announced the retirement of four members of its board of directors and named four new directors to replace them.
Apple on Tuesday reported a decline in revenue and earnings for its fiscal 2002 third quarter ended June 29.
Handspring has enlisted a sales and marketing outsourcer to help spur solution provider innovation on the Treo platform.
NetIQ's new SQL Management Suite 1.1, which is now being shipped, comes with two of its four component applications upgraded.
IBM is undertaking a multifaceted strategy to more closely integrate its WebSphere middleware with its integration software.
Microsoft is shipping two new editions of its BizTalk server aimed at enabling small and midsize companies to more affordably automate business processes and integrate B2B capabilities.
IBM has helped distributors Avnet Hall-Mark and Arrow Electronics' Support Net division launch programs for solution providers looking to reach into the lucrative life-sciences market.
Solution providers need to refocus, re-energize and rebuild to survive, even if that means a drastic change in their business models, GE Access executives told solution providers at the distributor's annual New Frontiers Conference.
Start-up RadioFrame Networks wants to open its unique pairing of wireless LAN and mobile telephone technologies to the channel, but solution providers say restrictions from the cellular industry could make selling the company's product a tricky proposition.
NTT/Verio introduced what it termed "lucrative" recurring commissions for partners Tuesday as part of a revamped channel program.
2015 Partner Programs Guide: 5-Star Emerging Vendors
As part of the Partner Program Guide, CRN designates some programs as 5-Star Partner Programs. Here are the 5-Star Emerging Vendors.
Top Ingram Micro Execs: What's Next For Its Cloud?
Ingram Micro leaders discussed everything from how commoditization threatens hosted cloud to why small businesses are most typically cloud pioneers to the continued need for on-premise infrastructure during the company's 2015 Cloud Summit in Phoenix.
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Toronto Rock beat Buffalo Bandits in NLL playoff preview
Kevin Ross, Rob Hellyer help lift Rock past John Tavares and Bandits at Air Canada Centre.
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Kevin Ross scored three goals and assisted on two more, leading the Toronto Rock over the Buffalo Bandits 13-9 in National Lacrosse League action Friday at the Air Canada Centre.
The Rock and Bandits will meet in the NLL playoffs, with home advantage yet to be determined.
Stephan Leblanc scored three and added an assist, Kasey Beirness had two goals and three helpers and Damon Edwards also scored two for Toronto (9-9). Rob Hellyer had a seven-point night with a goal and six assists, and Sandy Chapman and Scott Johnston chipped in with a goal apiece.
John Tavares led the Bandits (8-8) with two goals and two assists. Mark Steenhuis and Ryan Benesch had a goal and three assists apiece, Dhane Smith scored once and helped on two more and Joe Resetarits, Shawn Williams, Aaron Wilson and Mitch Jones also scored.
Rock goaltender Nick Rose made 48 saves in the win. Buffalo's Anthony Cosmo stopped 36 shots in the losing effort.
The Rock clinched a playoff spot last Friday by defeating the Philadelphia Wings 10-9 at the ACC. The team has made the playoffs for five consecutive seasons.
The Rock hold the third seed in the East Division and could still move up to second, ahead of the Bandits, and host their playoff date.
Before Friday night, the Rock had lost two in a row to the Bandits, both 12-10.
Five teams have clinched NLL playoff spots. In the East, the Rochester Knighthawks (12-4), Bandits and Rock are in. The Knighthawks have clinched the East title and home advantage through the first two rounds.
In the West, the Edmonton Rush (15-1) clinched first place overall while the Calgary Roughnecks (10-6) will be second. The Colorado Mammoth (6-10) and Vancouver Stealth (4-11) are fighting for the last spot.
With files from Curtis Rush
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Review Commonality of Strangers finds no neat conclusions at Nottingham's New Art ExchangeMahtab Hussain, a photographer who specialises in highlighting issues of multiculturalism, portrays the residents of Hyson Green in the gallery within the Nottingham community.04 February 2015
Preview Life, death, loss and escape in Nottingham with Zineb SediraFloating Coffins, a new video and sound installation by French-Algerian artist Zineb Sedira, explores life on the African coast and geographical barriers09 February 2009
News New Art Exchange and Artfinder ask public to turn curators in digital Culture Cloud experimentHaving been chosen from hundreds of applicants for Arts Council digital funding, the Nottingham gallery is teaming up with technological innovators ArtFinder to present a public art vote.15 June 2012
Preview Realism in Rawiya brings fresh perspectives on the Middle East to New Art ExchangeIn an exhibition in Nottingham, an all-female photography collective reveal alternative realities in the much-stereotyped Middle East.09 January 2013