text:
Foals edge past Augsburg
Mönchengladbach – Borussia Mönchengladbach moved up to sixth in the Bundesliga table with a laboured 1-0 win over FC Augsburg at Borussia Park.
Vogt's poor decision
After a fairly low-key opening, the contest flared into life midway through the first half when Augsburg's Kevin Vogt's was dismissed for a professional foul on Gladbach frontman Peniel Mlapa and Filip Daems tucked away the penalty. Thereafter, the hosts wasted a whole series of chances to put the result to bed, but coach Lucien Favre will nonetheless be happy with the points after last week's defeat in Stuttgart - as well as his team's first win over Augsburg since the Bavarian Swabians took their Bundesliga bow in 2011.
Luuk de Jong had scored three in his last six games for the Foals, but Favre opted to start instead with Mlapa in attack. It was the former Hoffenheim man's first league start in 13 months and it was a change that worked well for the home side.
After a quiet opening quarter, Mlapa it was who was scythed down by Vogt on 26 minutes, resulting in a red card for the defender and a penalty that Daems coolly dispatched to make it 1-0 - his eleventh successful conversion from as many attempts in the Bundesliga. Mlapa and Mike Hanke were seeing plenty of the ball, and minutes later it was almost 2-0 as a beautiful Hanke pass released his strike partner, but Mlapa's placed shot hit the post.
Augsburg had lost a large portion of their attacking threat when Tobias Werner was substituted in the aftermath of the red card, and they rarely threatened Marc-Andre ter Stegen in the opening half. The early moments of the second period also belonged to the hosts, with Juan Arango going close from a speculative free-kick. There was a certain lack of intensity to their play, however, which gave ten-man Augsburg hope.
The visitors' main strategy for getting close to the Gladbach goal was to win free-kicks in the opposing half, but they produced few results without the delivery of Werner. The home side fared a little better by using a similar technique, with Alvaro Dominguez glancing Arango's inswinger wide of the target. The game had lost all fluency by this point, however, with both sides committing a large number of fouls.
Chances go begging
A second goal would have killed the game for the hosts, but Hanke's volley struck the post, Branimir Hrgota headed over the bar and Granit Xhaka couldn't make the most of three good shooting opportunities. The Foals were then almost made to pay for their profligacy, as ter Stegen was called on to make a smart reflex save from substitute Knowledge Musona's prodded effort.
It was in the end a deserved win for Favre's men, but they will likely have to be more clinical to finish the season in one of the European places. Augsburg, meanwhile, remain in 16th place, three points ahead of Hoffenheim but having played a game more.
Line-ups and statistics
Bernie Reevies


text:
VHA and UHC to merge
February 4, 2015 in Medical Technology
By pooling their data and analytics capabilities, supply chain management firm VHA and academic medical center alliance UHC will help drive "unparalleled insight into improving the quality and total cost of care," VHA officials say.
The deal is expected to close in March. Financial terms have not been disclosed.
[See also: VHA gets bullish on population health]
Together, VHA and UHC will:
- Serve more than 5,200 health system members and affiliates
- Provide services to nearly 30 percent of the nation's hospitals, including nearly all the academic medical centers and health systems
- Serve more than 118,000 non-acute health care customers
- Have more than $50 billion in purchasing volume, the largest in the industry
- Provide services to all of the top 10 hospitals on the US News and World Reports annual list of America's Top Hospitals
- Have the industry's most in-depth clinical data combined with the nation's most robust supply chain data to address cost and quality
"VHA was founded on the idea that health care organizations perform better and faster when working together," said VHA President and CEO Curt Nonomaque, who will lead the new company. "This combined organization will have even greater ability to harness the power of health care's brightest minds to provide targeted expertise to improve financial, operational and clinical performance."
[See also: UHC inks RFID contract for its members]
Read more at Healthcare Finance News.
Article source: http://www.healthcareitnews.com/news/vha-uhc-merge

subset: commoncrawl


text:
Series FKHL Grippers have compact design and are ideal for various-sized work pieces. A dual piston mechanism synchronized by a rack & pinion delivers high grip force with great precision.
- Magnetic piston is standard
- Sensor slots integrated in body
- Oil impregnated sleeve bearings encased in steel are used for all shafts
- Rod scraper for dust protection is built in
- Fabco-Air


text:
After I finished Pimsleur, I knew I wanted a program that could help me with listening, reading, vocabulary development, and sentence variety. My textbooks just have audio of word lists and sample sentences and/or choppy, overly formal dialogues. Those serve their purpose in the learning process, but I need more, so I decided that I would subscribe to either Learn Thai Podcast or ThaiPod101 as those seemed to get the highest reviews. Both programs have trial materials available. As far as I can tell, Learn Thai Podcast only has beginning-level samples available for free whereas ThaiPod101 has a 7-day trial of their premium membership so you can check out all levels of their materials. Here's the rundown of why I chose Learn Thai Podcast:
The Price
Learn Thai Podcast requires a hefty initial investment of $197, which is why I put off signing up for so long. Yes, it's a five-year subscription, so it ends up being about $3.30/month, but it takes a bit of a leap of faith to invest that up front when all you can check out for free is the beginning materials. Luckily for me, I managed to get my subscription for just $97 because of a sale they had for the King's birthday. Now that I have it, though, I can see it is well worth the $197 they ask — it's actually probably worth much more. As an aside, there's no reason to fear that the price will increase anytime soon — the $197 "special offer" has existed about as long as the site.
ThaiPod101 not only costs more, but it has pretty abrasive advertising techniques like this one:
You have to scroll through pages of this before you get to the tiny "no thanks" link at the bottom so you can actually enter the site.
Once inside, you can see the different prices for memberships. The cheapest one is about 70 cents more per month than Learn Thai Podcast, assuming there are enough materials on ThaiPod101 to keep you busy for five years (it doesn't look like it to me).
Upon clicking the "upgrade" button on ThaiPod101, I found the prices are misleading. A premium membership is only $10 a month if you pay $240 at once and a Basic membership is only $4 a month if you pay $96 at once. If you want to pay month by month, memberships are $25 and $8 per month, respectively.
Unfortunately, the best features of ThaiPod101 are only available to Premium members, and I don't think there are enough materials at this pount to justify a two-year membership (more on that below).
Audio Quality
In this section, I'm only talking about audio clarity. On this front, ThaiPod101 has more consistently clear audio. All of their audio appears to have been recorded in a studio and has no background noise (other than intentional sound effects) at all. Learn Thai Podcast, however, takes their authentic conversation audio and video (the aspect I am most interested in) from various sources, including handheld cameras on a windy day. It's a bit rough on the untrained ear. After each conversation the Thai voice of the podcast (Jo?) speaks the phrases again with studio-quality sound. Their grammar and vocabulary lessons are perfectly clear.
Interest Level of Materials
Thai Pod 101 has a 7-day premium trial where you have access to all their materials, so I was able to listen to beginning, intermediate, and advanced recordings. The English introductions and discussions after were so dry and dull I couldn't handle it. Based on my husband's wincing during the Thai portions, my guess is the same is true for the Thai sections. My goal for learning Thai isn't to sound like a monotone textbook, and the lack of phrasing by the American speaking Thai (in all authentic Thai I've heard, there are longer spaces before/after certain words to separate phrases and clauses — these recordings seemed to put an equal space between each word) was distressing. The native Thai speaker spoke well, but it seemed like the non-native actually got more speaking time on most dialogues. Also, for dialogues among three or more people, the two speakers tried to disguise their voices. The male (American) speaker, when being a third person, sounded like Frank Oz trying to be somewhere in between Yoda and Fozzie Bear. The result was painful to listen to. On the plus side, the material, particularly at the more advanced levels, talked about important parts of Thai culture. That would have kept my interest despite the monotony if the information weren't presented in the form of a second grade book report.
Learn Thai Podcast, on the other hand, has conversations and monologues that are husband-approved in their authenticity. In fact, Learn Thai Podcast is the only learning resource I have that hasn't made my husband double over in laughter. When I showed him some of the intermediate and advanced dialogues and grammar lessons, he was pretty impressed.
Range of Materials
This is where Learn Thai Podcast completely blows ThaiPod out of the water. Look at this:
That's seventy-three intermediate lesson cycles alone! Not to mention the six advanced cycles, five advanced news cycles (note each cycle is four lessons), sixty beginner grammar, review, and bonus lessons, 317 beginner vocabulary lessons (each with about ten words). I had heard Learn Thai Podcast had a lot of lessons, but I didn't expect it might actually keep me busy for the next five years.
ThaiPod101 has a good amount, too, especially in the advanced audio blog section (there are about 50 lessons there), but it simply doesn't even approach the number of lessons Learn Thai Podcast has.
Quality of Materials
The fact is, I'm not going to study with something that bores me, and ThaiPod101 is painfully boring. My toaster has more emotion than those speakers, and the banter is some of the worst I have ever heard. It has frills — opening music, sound effects — but I don't need frills, I need speech that sounds like human beings. I couldn't even pay attention to the explanation of the phrases in the dialogues. Bad, bad, bad.
Learn Thai Podcast has no frills at all. It's low-budget and straight to the point, just the way I like. Plus, the explanations of the dialogues and the number of repetitions are perfect. I look forward to studying with Learn Thai Podcast, even if it's just the deadly themed vocabulary lists (something ThaiPod101 has managed to make even more boring than usual). Most importantly, I'm learning. I understand so much more of what my in-laws say than before, and I'm getting a better grasp of how to string words together.
Reading Support
Both Learn Thai Podcast and ThaiPod101 have transcripts of all their audio materials in both Thai script and romanization, and both have flashcard features for words and phrases. Both provide Thai script to read along with as you listen. For Learn Thai Podcast, the transliteration is far enough below the Thai that it can easily be covered up or ignored if you're like me and think transliteration is a tool of the devil. They also have pdf files of the transcriptions. ThaiPod101 has dialogue transcriptions on the screen below the audio player as well as pdf files in English, Thai, and transliterated Thai. I would say both programs are equal here.
As far as learning how to read Thai script, Learn Thai Podcast has a section for that that I haven't yet explored since I can already decode. ThaiPod101 has an alphabet page, but I couldn't find anything on actually learning how to read.
Visual Appeal and Ease of Use
I think both are equally easy to navigate and both are equally visually appealing.
The Final Decision
Because of ThaiPod101's aggressive and misleading marketing practices and their use of non-native speakers who speak like robots, I found Learn Thai Podcast to be more valuable for me. If ThaiPod101 were to be more upfront about pricing and/or charge $5 a month for Premium, I would consider a two-year membership because the advanced blogs, though dull, are pretty decent listening practice and it's hard to find extended texts with audio. As it is, though, they've made a bad impression on me as a company.
UPDATE: Customer Service
Since writing this post, I've e-mailed both Learn Thai Podcast and ThaiPod101 with questions about their features. Both companies responded to me within 24-hours with very helpful responses. I had read elsewhere that ThaiPod101 customer service isn't great, but I have to strongly disagree — they were excellent! It actually makes me feel a little more forgiving about their marketing techniques and the stodgy acting on the beginner-level podcasts. They reset my 7-day trial, so I think I'll do a review later his week on the specific features of each program so people can make informed choices. Basically, I'm trying to write the reviews I wish had been available when I was trying to choose between the two.


text:
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Researchers likely won't have to chase after enrolled study participants to sign new consent agreements if revised human subject protection regulations apply, an HHS office head told Bloomberg Law.
If researchers choose to implement options designed to reduce burdens associated with the revised Common Rule, new consent requirements would be triggered. The Common Rule is a set of regulations adopted by nearly 20 federal agencies to protect human research subjects from undue harm (45 C.F.R. 46).
However, Jerry Menikoff, director of the Health and Human Services Office for Human Research Protections, told Bloomberg Law he doesn't expect new consent requirements to apply to people who have enrolled in a study before Jan. 21. The revisions take effect on that date, and the burden-reducing options apply from July 19.
"We're not saying you have to go back and re-consent people," Menikoff told Bloomberg Law in a July 20 interview. But he stressed the OHRP hasn't published any official notices or guidance on this matter. "This is part of the guidance that's not out yet. But there's a sense that the guidance would probably go in that direction."
July 19 kicked off a six-month transition period, during which research institutions can adopt the three burden-reducing provisions. But taking this option means an institution has decided to transition that study onto the revised Common Rule, and all the new requirements kick in Jan. 21. If an institution doesn't use the burden-reducing provisions, that study can continue to comply with the old rule.
Reducing burdens was one of the two main goals for updating the Common Rule from the 1991 version, along with updating the regulations to better align them with modern research practices. But with two delays to the start date, the revised rule has undergone a bumpy rollout. The past year and a half has left many institutions unclear about how to execute new requirements.
"The question is whether or not you're going to save burden at the end," David Borasky, a North Carolina-based research compliance executive who routinely handles these issues, told Bloomberg Law in a July 20 interview.
The burden-reducing provisions cover: a new definition of research that reduces the number of activities subject to the regulations; elimination of an annual checkup known as continuing review for some low-risk studies; and elimination of a requirement that ethics panels known as institutional review boards review grant applications.
If a study met the criteria for an expedited review—which qualifies it for the burden-reducing provision to end continuing review—doing away with the continuing review requirement could trigger a need to rewrite the consent form next January.
"Assuming they're enrolling new subjects from January onward, for those subjects, it's probably likely you'd have to revise the consent form in some way or another," Menikoff said.
Consent requirements are the main part of the new Common Rule that would require studies to go back to an IRB for approval, said Borasky, who is vice president of IRB compliance for WIRB-Copernicus Group, a commercial provider of regulatory and ethical review services for clinical research. While it would be nice to eliminate some continuing review, Borasky said, it's not a substantial burden.
The same goes for the elimination of IRB grant review, he said. "This is not the thing that makes or breaks an IRB office" in terms of managing their human research protection programs, Borasky said. "It'll be nice not to have to do it when the whole rule goes into effect."
A research project that has reached the point of data analysis also could qualify for elimination of continuing review, and Menikoff said those study participants wouldn't need to be re-consented. Use of this provision might make more sense in this situation. "Those are studies that have less of a downside in terms of making a decision to transition them," he said.
The HHS generally doesn't comment on the status of guidances that haven't been released publicly, but Menikoff said the OHRP is working on a number of draft documents to help institutions gear up for the new revisions. He said the office will factor in public comments when rolling them out, such as the call for more guidance on the key information requirement in consent forms.
All Bloomberg BNA treatises are available on standing order, which ensures you will always receive the most current edition of the book or supplement of the title you have ordered from Bloomberg BNA's book division. As soon as a new supplement or edition is published (usually annually) for a title you've previously purchased and requested to be placed on standing order, we'll ship it to you to review for 30 days without any obligation. During this period, you can either (a) honor the invoice and receive a 5% discount (in addition to any other discounts you may qualify for) off the then-current price of the update, plus shipping and handling or (b) return the book(s), in which case, your invoice will be cancelled upon receipt of the book(s). Call us for a prepaid UPS label for your return. It's as simple and easy as that. Most importantly, standing orders mean you will never have to worry about the timeliness of the information you're relying on. And, you may discontinue standing orders at any time by contacting us at 1.800.960.1220 or by sending an email to email@example.com.
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text:
University of Maryland
CMSC 422: Introduction to Machine Learning (Spring 2012)
Tuesdays and Thursdays, 11 am; Room CSI 3120
Instructor: Jim Reggia
The field of machine learning has been growing very rapidly during the last decade, reflecting
the successful application of increasingly powerful computational techniques and the expanded
opportunities for data mining brought about by large amounts of online data. This course provides
a broad overview of existing methods for machine learning and an introduction to adaptive systems
in general. By the end of the semester students should be familiar with the central concepts and
most commonly used methods in this field.
Important Dates
Assigned Readings
Homework Assignments
Online Resources
Web Accessibility


text:
The 2015 edition of Connected Automobiles willtake place on October 28-30 in Monza F1 Circuit (Milan – Italy), a charming location and a worldwide recognized symbol of innovation in automotive. The exclusive international conference Connected Automobiles is part of Smart Mobility World: the European reference event for the sustainable, connected and integrated mobility of the XXI century – featuring two days of conference and networking initiatives. In 2014 Smart Mobility World counted over 3.500 visitors and 152 partners, sponsors and exhibitors. The matchmaking program featured 119 b2b meetings among 116 participants from 19 countries.
Our Partners - India Smart Automotive Forum 2015
- m2m2iotpaper.com
- ABI Research
- Ajeenkya DY Patil University, Pune
- Auto & Design
- Business with India
- Clocate.com
- Connected Automobiles 2015
- Connected Vehicle Trade Association (CVTA)
- Council Global
- Datafloq
- Department of Electronics and Information Technology, Ministry of Communications and Information Technology, Government of India
- Digital India
- India Electronics & Semiconductor Association (IESA)
- Innovability S.r.l.
- Karunya University
- M2M Asia Pacific
- Mind Commerce
- RE news india
- Robohub
- Smart Webzine
- Symbiosis Institute of Telecom Management (SITM)
- Technology Information Forecasting and Assessment Council (TIFAC)
- Telecompaper
- TelecomWatch
- The School of Planning and Architecture (SPA)
- The Trusted Computing Group (TCG)
- WSNBuzz


text:
The words poured effortlessly from my fingertips and onto the page completely free of grammatical errors and typos, forming a perfect, succinct, and intriguing story that everyone in the world wanted to read…said no author ever. Rewriting and editing comes with the job and there's absolutely no way around it, but luckily there are ways to make it less painful.
Since finishing The Nightmare Birds, I've already re-read the entire thing a half dozen times on my laptop, then another two times printed out, then passed it off to a few beta readers, re-read it again, and then sent it off to my editors for a final polish. When I get it back, I'll have more editing, more rewriting. Then, there will be another reading or two printed out and then a final arc read through. It takes forever and is incredibly tedious, but it's well worth it, especially if you compare what you originally wrote to the final thing. Just take a look at this massacred page from one of my Strange Luck edits.
My first drafts are terrible. They're slow and peppered with lots of unnecessary scenes and words. I also have a tendency to think a particular word, but type a similar sounding word with a completely different meaning instead. It's super fun going back and finding those (*eye roll). The worst is when you think you've finally tamed the beast, but you open the Word doc to find it covered in red.
If there's one piece of solid gold advice I've learned from rewriting and editing over the years it's this: TAKE BREAKS FROM YOUR STORY. As soon as it starts to feel like "work," becomes boring, you stop looking forward to working on it, things stop making sense, and/or you begin losing track of important details (dates, ages, hair color, etc.), then it's time to put the manuscript down and walk away. I know it may seem counter-intuitive because you're in "the zone," but trust me, you'll end up doing more harm than good. I've had to force myself to walk away from The Nightmare Birds probably a dozen times to recollect myself. Sometimes it was for a few days, other times a few weeks. This has meant pushing back my launch date, which has really sucked, but I'd rather have a well-written book that comes out a little later than a garbled, incoherent story.
Even if you're not experiencing any of the symptoms above, YOU STILL NEED TO TAKE BREAKS from your work. Only you will know when and for how long. Just make sure it's at least a few times.
|This is totally me. :P
Why get up and walk away? Because you'll be able to look at your story with fresh eyes. Taking a break also rekindles the passion for your story and characters. It took me a long time to implement this practice, even after I had read Neil Gaiman's quote when I first started writing seriously:
"The best advice I can give on this is, once it's done, to put it away until you can read it with new eyes. Finish the short story, print it out, then put it in a drawer and write other things. When you're ready, pick it up and read it, as if you've never read it before. If there are things you aren't satisfied with as a reader, go in and fix them as a writer: that's revision."Walking away is one of the most critical things you can do as a writer, but it's not to say that you shouldn't write at all, just go write something else. Anything else. Your characters will still be there when you return. Your lumpy desk chair will still be there when you return. Your keyboard peppered with food crumbs will still there. I promise. The point is that you need to clear your head so that when you return you'll be at your absolute best.
How do you handle edits? How painful is rewriting for you? Share your tips, tricks, and advice below.
Have you subscribed yet??? It seriously makes my day when I get a new subscriber. :)


text:
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text:
This plugin is an add-on for the Gravity Forms plugin. If you don't yet own a license for Gravity Forms – buy one now! (affiliate link)
What does this plugin do?
- make a list field column accept only numbers
- specify the format of the number including currency, comma delimited (9,999), dot delimited (9.999) and no formatting (9999)
- specify the rounding type – round up, round down or round closest
- specify the decimal places to round to – no rounding, 0, 1, 2, 3, 4, 5
- force the number to use fixed placed notation – e.g. 10.1 would become 10.10 with 2 place fixed notation
- add a column total – automatically display the total of the column
- specify a range requirement – this can be a set number, e.g. 200 or a formula, e.g. column 1 + column 2
- calculate column values uing a formula (e.g. field = column 1 + column 2)
- compatible with Gravity Forms List Field Calculations Add-On
- compatible with Gravity PDF
See a demo of this plugin at demo.itsupportguides.com/list-field-number-format-for-gravity-forms/
How to I use the plugin?
Simply install and activate the plugin – no configuration required.
Open your Gravity Form, edit a 'List' field and use the 'Number Format' options to configure the columns.
Have a suggestion, comment or request?
Please leave a detailed message on the support tab.
Let me know what you think
Please take the time to review the plugin. Your feedback is important and will help me understand the value of this plugin.
Gravity Forms is a trademark of Rocketgenius, Inc.
This plugins is provided "as is" without warranty of any kind, expressed or implied. The author shall not be liable for any damages, including but not limited to, direct, indirect, special, incidental or consequential damages or losses that occur out of the use or inability to use the plugin.
Note: When Gravity Forms isn't installed and you activate this plugin we display a notice that includes an affiliate link to their website.
- Install plugin from WordPress administration or upload folder to the
- Activate the plugin through the 'Plugins' menu in the WordPress administration
- Open the Gravity Forms 'Forms' menu
- Open the forms editor for the form you want to change
- Add or open an existing list field
- With multiple columns enabled you will see a 'Number Format' section – here you can choose which columns are number fields.
- How do I base a calculation from a drop down (select) field?
This plugin is capable of making calculations across a row using drop down (select) fields.
To do this you first need to set a column in the list field as a drop down – I suggest you use the Drop Down Options in List Fields for Gravity Forms plugin for this.
You then need to make sure the VALUE (not the text label) for the drop down is or contains the number for the calculation.
Once you have done this simply 'number enable' the column and setup the total calculation for the destination column.
GLAD: Numeric List Fix Support
Awesome Plugin
Great Add-on
Awesome add-on
Contributors & Developers
"List Field Number Format for Gravity Forms" is open source software. The following people have contributed to this plugin.Contributors
Translate "List Field Number Format for Gravity Forms" into your language.
Interested in development?
Browse the code, check out the SVN repository, or subscribe to the development log by RSS.
*Fix: resolve conflict with Gutenberg – adding form using a shortcode would give an error message
*Fix: resolve issue with not being able to add new list rows in wp-admin entry editor
*Fix: resolve issue with column total appearing when column was previous 'number enabled'
*Fix: resolve issue with single column list field 'Enable Column Total' not displaying the set state
*Fix: resolve error when submit 'Warning: A non-numeric value encountered in list-field-number-format-for-gravity-forms/list-field-number-format-for-gravity-forms.php on line 590'
- Fix: resolve issue with column total not display in wp-admin entry view
- Feature: add no formatting (9999) option – e.g. a year "2019"
- Fix: number preview in form edit wasnt applying the correct formatting
- Fix: resolve issue with the total row not calculating in the backend entry editor.
- Maintenance: resolve 'undefined index' error message when debug is enabled.
- Feature: add support for pre-filled list fields (gform_field_value_ filter)
- Fix: Resolve issue with column totals repeating/cascading in entry detail and emails.
- Fix: Resolve issue with blank row being added to list field when viewing entry details.
- Fix: Improve Gravity Flow support – allow Gravity Flow to output the correct value/format depending on the form editable status.
- Fix: Resolve compatibility issue with Gravity Forms List Field Calculations Add-On. If a list field had multiple total rows and the columns were being used to calculate separate number fields – the calculation would include the total column value.
- Feature: add ability to base calculations off of drop down (select) fields in list field row. See plugin FAQ for more information.
- Fix: resolve conflict with Drop Down Options in List Fields for Gravity Forms plugin
- Maintenance: improve JavaScript to better handle calculations that include values coming from non-list fields.
- Fix: resolve conflict with Ajax Upload for Gravity Forms plugin (PDF would not display correctly when a list field contained both an 'Ajax Upload' column and a 'number format' column.
- Maintenance: change how plugin checks for Gravity Forms being installed and active
- Fix: resolve 'Fatal error: Can't use function return value in write context' error message for older versions of PHP (5.4 or earlier)
- Fix: improve validation for 'max' and 'min'
- Fix: add Gravity PDF support for 'number enabled' single column list fields
- Maintenance: improve validation for French notation numbers (9.999,99)
- Fix: resolve 'undefined variable' error on submitting form.
- Fix: resolve issue with total column repeating if there are multiple total columns in a list field
- Maintenance: improve how rounding and fixed point notation is handled in JavaScript and PHP
- Feature: allow calculations to be made from standard Gravity Forms number fields in addition to list field column fields
- Fix: resolve issue that stopped negative values from being used, for example -50 would be treated as 50
- Maintenance: improve support for 'ABN Lookup for Gravity Forms' plugin
- Maintenance: improve support for 'Sortable List Fields for Gravity Forms' plugin
- Fix: JavaScript improvements.
- Fix: Improve JavaScript in entry editor.
- Fix: Change how field id is determined to improve compatibility with all versions of Gravity Forms.
- Fix: Resolve issue with row calculations based on French notation numbers (9.999,99).
- Fix: Resolve issue field format not correctly setting when new row is added.
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- Feature: calculate column values uing a formula (e.g. field = column 1 + column 2)
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- Feature: add client-side and server-side range validation
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- First public release.


text:
Canticle to the Cosmos Series: 12 - 60 minute programs/$240.00 for the set.
Brain Swimme, BS, PhD., featured in the BBS series Soul of the Universe along with Stephen Hawkins, hosts this classic series on the New Story of the Universe. Designed to be used as part of an academic curriculum, in small group study, or for individual enrichment. Item #CS002.
1. The Story of Our Time
2. The Primeval Fireball
3. Feast of Consciousness
4. Fundamental Order of the Universe
5. Destruction and Loss
6. A Magical Planet
7. Sex, Death & Dreams
8. The Nature of the Human
9. Fire in the Mind
10. The Timing of Creativity
11. The Human Story
12. A New Prosperity
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The Hidden Heart of the Cosmos - 80 minutes/$59.95
What does it mean to be human, to be alive on planet Earth in the midst of the vast universe as it is now understood? Cosmologist Brian Swimme takes us on an exhilarating journey in search of the new story that is developing in answer to this question. From the Milky Way to the Virgo Cluster of galaxies. Drawing on 20th century discoveries in quantum physics and cosmology, he presents a stunning perspective. 31/2 Stars - Highly Recommended. Video Librarian. Inspiring to teachers, college classes and the philosophically inclined. Booklist. Winner - Bronze Apple - National Educational Media Award 1996. #CS001
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Dog Grooming: How-To Guide - 60 minutes/$19.95
Provides step-by-step instructions for the pet owner. Learn how-to use brushes and combs effectively; properly bathe; clean ears; brush teeth; clip nails; mix special for flea dipping and much more. Keep your dog wellgroomed, healthy and happy. "A well-produced, comprehensive package . . . excellent job . . . a treat for dog lover" - Video File. Item #CG009
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How to Save Big Money When Buying a Car - 95 minutes/$39.95
Now you can: learn the 5 biggest buying mistakes and how not to make them; avoid deceptive sales practices; know the most important question to ask the salesperson; negotiate your own terms; learn financing strategies, best time to buy, from whom to buy and much more. Features Ed Madden, former car salesman and Consumer Advocate with the Illinois Attorney General's Office. A comprehensive and informative process that would prove helpful to and consumer. Mary Clark, Chicago Public Library. A very thorough presentation. Superb job and highly recommended for public libraries. Judy Napier, Schaumburg Library District. Item #BS002.
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Making Wine at Home the Professional Way - 76 minutes/$29.95
A step-by-step look: from grape picking to crushing, fermenting, pressing, racking and bottling - all presented in an entertaining, easy-to-understand way. Lance Cutler, who has a Master's Degree in Curriculum Development, designed the video to teach the complexities of winemaking in a simple, straight-forward way. His style of instruction - irreverent, funny and informative - lends to the learning process, while peaking the viewer's interest. The process is clearly outlined - Booklist. 3 Stars - Video Librarian. Item #WP001
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Plantings & Growth - 30 minutes/$14.95
As seen on TV, Planting & Growth is the complete do-it-yourself guide to creating a more pleasing and bountiful outdoor living enviroment. Item #BHVE01
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Stan Nicotera - e-mail:
Special Interest Video Sales Group, PO Box 968, Englewood, FL 34295
phone 941-473-2601 - fax 941-473-2701


text:
It's wrong that you just apply sun-proof products at the day of high exposure. Do you know even in the dark weather, UV radiation rays in the sunlight remain active which most cause wrinkles, age spots, fragility, etc. Over the years, your skin naturally shows signs of aging and the worse cause is the accumulation of sun radiation. As the result, there are some effective ways to sweep out skin damages.
Appropriately applying sun-proof cream
Remember to apply sun-proof cream 30 minutes before going out and redo after each 90 minutes. Especially, in the water you should rub sun-proof more regularly with a adequate quantity. The ideal index of sun-proof cream is SPF 15 upwards.
Assuring skin moisture
Moisture lack is the culprit of crack skin, so it's vital to apply moisturizing cream. You should drink much water and apply moisturizing cream two time each day. Sun-proof cream containing moisturizing substance is the best.
Simplifying make-up
It's advised that you shouldn't abuse make-up, please mildly make up. You know in most of the skin care products, especially in make-up cosmetics, there are lots of substances highly exposing to the sunlight. Therefore, you should simplify your make-up steps only with eye-lining, lipstick and base- cream applying.
Applying dead-skin removal cream
To restore the skin's appearance, applying dead-skin removal cream 2-3 times every week will promote the effect of skin care products. It's better to remove dead cells with natural products such as fruits and plants.
Diet balance
Remember to maintaining the daily diet with anti-oxidized green foods such as hami melon or avocado. Besides, eggs and fishes help effectively improve your skin.


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Раздел: root - Linux - Squid
squid под высокой нагрузкой. [частично решено]
Can_I_make_Squid_proxy_only. Without_caching_anything.
Sure, there are few things you can do.
You can use the cache access list to make Squid never cache any response:
cache deny all
With Squid-2.7, Squid-3.1 and later you can also remove all 'cache_dir' options from your squid.conf to avoid having a cache directory.
With Squid-2.4, 2.5, 2.6, and 3.0 you need to use the "null" storage module:
cache_dir null /tmp
Note: a null cache_dir does not disable caching, but it does save you from creating a cache structure if you have disabled caching with cache. The directory (e.g., /tmp) must exist so that squid can chdir to it, unless you also use the coredump_dir option.
To configure Squid for the "null" storage module, specify it on the configure command line:


text:
Knoxville, TN (PRWEB) February 11, 2016
Brookhaven Retreat ® LLC, a unique residential treatment facility exclusively for women with mental health and/or substance abuse issues, will exhibit at the psychiatry event of the year, the 2016 annual meeting of the American Psychiatric Association at the Georgia World Congress Center in Atlanta, GA from May 14 to 18.
Brookhaven Retreat offers a unique voluntary residential program of mental health and substance abuse services designed to help women overcome depression, trauma, anxiety, personality disorders, and substance dependence among other diagnoses.
Jacqueline Dawes, owner and founder of Brookhaven Retreat, and her team will be at the 20x10 exhibitor booth #1513 answering questions about The Lily Program®, a 90-day individualized mental health treatment program offered exclusively at the Tennessee-based treatment center. Visitors of the booth can enter a drawing to win prizes and receive a messenger bag with information about BH and additional giveaways.
Originally established in 1844, the meeting has remained the premier gathering in psychiatry today and serves as a symbol of the Association's commitment to improved patient care through professional education. Hundreds of educational sessions and presentations on the latest research will be featured. More than 13,000 attendees from around the globe, including physicians from psychiatric and other mental health disciplines will gather at the Georgia World Congress Center, the first state-owned convention center established in the United States, and now the third largest convention center in the country.
Attendees will gain insight and in-depth knowledge designed to help improve patient care by meeting with renowned experts, earning credits, building peer relationships, discovering ground-breaking technology and new therapies and treatment options.
"Each year we look forward to this meeting as an opportunity to mix with peers, gain insight in our field, and for an up-close and more personal view of what new concepts, research and information have been developed and are in the process of being developed," says Dawes. "It also affords us the very important chance to introduce ourselves to those who don't already know who we are and what we do."
About Brookhaven Retreat
Brookhaven Retreat is a women's treatment center nestled on a naturally beautiful 48-acre site secluded in the foothills of the Great Smoky Mountains. It has helped hundreds of women across the United States overcome depression, trauma, anxiety, substance use and a range of other behavioral health challenges. Brookhaven's Founder, Jacqueline Dawes, has predicated its gender-specific treatment on "healing emotional breakage" for women. In this way, she has established a sanctuary and a place where women can feel safe, secure and cared for by a staff of highly trained professionals.


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- Research
- Open Access
A transferable heterogeneous two-hybrid system in Escherichia coli based on polyhydroxyalkanoates synthesis regulatory protein PhaR
© Wang et al; licensee BioMed Central Ltd. 2011
- Received: 28 February 2011
- Accepted: 9 April 2011
- Published: 9 April 2011
Polyhydroxyalkanoate (PHA) synthesis regulatory protein PhaR contains a DNA binding domain (DBD) and a PHA granule binding domain (GBD), it anchors to the promoter region of PHA granule-associated protein (PhaP) to repress phaP expression. However, PhaR will bind to PHB granules and be released from phaP promoter region when PHA granules are formed in vivo, initiating expression of phaP gene. Based on this regulatory mechanism, a bacterial two-hybrid system was developed: PhaR was separated into two parts: DBD was used to fuse with the bait, GBD with the prey, and phaP was replaced by a reporter gene lacZ. However, GBD protein expressed in vivo formed inclusion bodies. Thus, PhaP with strong binding ability to PHB granules was employed to replace GBD.
Three model interaction partners bFos, bJun and bATF2 were used to study the feasibility of this bacterial two-hybrid system compared with the controls lacking one or more essential elements of this system. Results showed that bFos, bJun and bATF2 bound tightly in pairs to allow strong expression of β-galactosidase in different expression levels. In contrast, very weak β-galactosidase activity was detected in all control groups.
β-Galactosidase activity level precisely correlated with the interaction force of tested protein pairs, and very weak β-galactosidase expression was detected throughout the control groups, which demonstrated the feasibility of this system for studying protein interactions.
- LacZ Expression
- Interact Protein Pair
- LacZ Activity
- Copy Number Vector
- Strong Binding Ability
Protein-protein interactions (PPIs) are essential in virtually all biological processes [1]. In the past two decades, a number of technologies to identify interacting proteins or to study these interactions have been extensively developed [2–6]. Among them, the most widely and successfully used methodology is the yeast two-hybrid system, originally developed by Chien et al [3], it exploits hybrid genes to detect protein-protein interactions by means of expression activation of a reporter gene [4]. Recently, a number of bacterial-based hybrid systems have been studied and become widely used. So far, studies of protein interactions in bacteria have centered on fusions to transcriptional repressors such as λcI, LexA or AraC, transcriptional activators involving the recruitment of RNA polymerase or the dimerization of the Vibrio cholera ToxR, complementation of biosynthetic enzymes such as dihydrofolate reductase, or signaling enzymes, e.g., the Bordetella pertussis adenylate cyclase [5]. By contrast, bacteria-based systems present advantages over yeast-based technologies, such as lack of cellular compartmentalization, faster growth and higher transformation efficiencies that are attainable permitting rapid and more efficient screening of complex libraries [6]. In spite of these advantages, all bacterial strategies have their drawbacks, including the need to employ the host intrinsic proteins for strategies of enzyme complementation, which will result in possible false positive outcomes, so host self-existed enzymes should be deleted from the host genome to eliminate interferences. This increases the complexity of genetic manipulation, and such technology developed could not be transferred to other bacterial strains. Therefore, a simple and transferable hybrid strategy should be developed to meet rising experimental demands.
Fos, Jun and ATF2 are transcription factors with basic leucine zipper (bZIP) domain. The relative dimerization efficiencies among the bZIP domains of the Fos (bFos), Jun (bJun) and ATF2 (bATF2) have been determined by the multicolour bimolecular fluorescence complementation (BiFC) assay [13]. Apparently, the bZIP domains of Fos, Jun and ATF2 can interact with each other in all pairwise combinations in vitro and in vivo [14]. In living cells, bJun:bFos heterodimers form more efficiently than either bFos:bATF2 or bJun:bATF2 heterodimers. Moreover, bJun:bFos heterodimers and bJun:bJun homodimers can coexist in cells with a content ratio of 60%:35% [15].
In this investigation, PhaR was intended to be developed into a platform for protein interaction study. To achieve this, PhaR was originally divided into two parts, namely, DBD and GBD. DBD fused to a bait protein, GBD fused to a prey protein, and phaP gene under the control of its native promoter was replaced by a reporter gene lacZ (Figure 1B). If the bait and the prey interact with each other, the new DBD-bait:prey-GBD complex will drop from the phaP promoter region, and LacZ will be expressed, with its expression level indicating the interaction strength of the bait and the prey. However, GBD protein expressed in vivo formed an inclusion body probably due to its high contents of hydrophobic amino acid residues. To avoid this, PhaP also having strong binding ability to PHB granule was employed to replace GBD (Figure 1C). Also in this study, three model interaction partners, bFos, bJun and bATF2, were used to test the feasibility of this system. Pairwise interactions among these proteins leaded to different levels of LacZ expression, directly reflecting their factual interaction strengths [15]. Since all elements involved including PHB synthesis operon phaCAB, DBD, phaP and phaP promoter are extrinsic in E. coli, false positive outcomes could be avoided. Moreover, the plasmids harboring these elements can be transferred to other appropriate host organisms for specific purpose, such as post-translational modification and glycosylation, allowing the heterogeneous two-hybrid system to become more extendable.
Materials and methods
Conserved domain prediction of PhaR
The conserved domain prediction of PhaR (GenBank: YP_725943) was conducted using Pfam software http://pfam.sanger.ac.uk/search/sequence. Results showed that the DNA-binding domain was in the N-terminal region (10th to 73th). To retain full DNA-binding ability, the 1st to 106th amino acid segment of PhaR as DBD was chosen for this study [12].
Bacterial strains and plasmids
Bacterial strains and plasmids used in this study
Strains & plasmids
Source or Reference
E. coli XL1-Blue
endA1 gyrA96 (nalR) thi-1 recA1 relA1 lac glnV44 F'[::Tn10 proAB+ lacIq Δ(lacZ) M15] hsdR17 (rK- mK+)
R. eutropha H16
Wild type (PHB+)
Ampr, pBluescript SK-, harboring PHB synthesis operon from R. eutropha (phaCAB operon), used to produce PHB granules
Ampr , pBHR68:phaP under control of phaCAB promoter
This study
Ampr , pBHR68: bFos-phaP under control of phaCAB promoter
This study
Ampr , pBHR68: bJun-phaP under control of phaCAB promoter
This study
Ampr , pBHR68: bATF2-phaP under control of phaCAB promoter
This study
Ampr , pBHR68Δ(phaCAB):bFos under control of phaCAB promoter
This study
Ampr , pBHR68Δ(phaCAB): bFos-phaP under control of phaCAB promoter
This study
Cmr, Tetr, cloning vector
Cmr, pACYC184:lacZ under control of phaP promoter
This study
Cmr, pOZ:DBD-bJun under control of phaCAB promoter
This study
Cmr, pOZ:DBD under control of phaCAB promoter
This study
Ampr, pTWIN2:phaP-intein-lacZ
Plasmid designs
Several genes were involved in this bacterial two-hybrid system including DBD sequence, phaP, lacZ under phaP promoter and PHB synthesis operon phaCAB. In this study, two compatible vectors, pACYC184 and pBHR68 were chosen to harbor related genes. DBD sequence and lacZ under phaP promoter were cloned to pACYC184, a low copy number vector (Additional file 1, Fig. S6). PhaP gene and PHB synthesis operon phaCAB were cloned to pBHR68, a high copy number vector (Additional file 1, Fig. S2). This aimed to achieve an excessive expression of PhaP (much more than DBD), so that all DBD-bait fusions had full access to prey-PhaP fusion. Simultaneously, a high copy number of PHB synthesis operon may help to produce more PHB granules. Sufficient PHB granules should ensure adequate space for attachment of all expressed PhaP or its fusion proteins. This is supported by previous study [20], after the PHB granules were removed from crude cell extracts via mild centrifugation, no PhaP was detected in culture supernatants using Western Blotting, indicating that all PhaP expressed in vivo were attached on PHB granules. These results pointed to an ideal situation: as long as the interaction of bait and prey is strong enough, all DBD-bait proteins can be caught by prey-PhaP to form a DBD-bait:prey-PhaP complex, which became attached to PHB granules, leading to the release of DNA repression on LacZ expression. Therefore, the expression level of LacZ depends on the interaction strengths of bait and prey.
Cultivation of strains, protein expression and PHB production
For the purpose of gene clone, recombinants of E. coli XL1-Blue were grown at 37°C and 200 rpm (FUMA QYC2112, Shanghai, China) in Luria-Bertani medium containing 1% w/v Bacto tryptone, 0.5% w/v yeast extract and 1% w/v NaCl overnight. When needed, tetracycline (50 μg/ml) or ampicillin (100 μg/ml) or chloramphenicol (34 μg/ml) was added to the medium. For the sake of protein expression and PHB granules production, the recombinants of E. coli XL1-Blue were cultivated at 37°C and 200 rpm (FUMA QYC2112, Shanghai, China) for 14 h in Luria-Bertani medium supplemented with 20 g/L glucose, to ensure sufficient PHB granules accumulation for this study as was also reported [21].
Assays of β-galactosidase activity
Where, according to the manufacturer's protocol, "1.7" represents total reaction solution volume (ml), "0.1" sample volume (ml), "0.0045" is molar absorption coefficient of o-Nitrophenol at 420 nm (ml/nmol/cm), "T" stands for the reaction time (min): the reaction is defined as the time consumed started from placing the reaction solution into 37°C thermostatic water-bath to the change of solution color to light yellow, "1" reflects the light path length (cm), "C" the protein concentrations of samples (mg/ml). Each value represents the average of three parallel samples.
Statistical Analysis
The statistical significance was evaluated by Duncan multiple range test [22], which was used to perform analysis of significant differences for β-galactosidase activity data. Probability values of p < 0.01 were interpreted as denoting statistical significance. Statistical analyses were performed by PRISM software (GraphPad, San Diego).
Identification of DNA binding domain and PHB granule binding domain of PhaR
Study of the two-hybrid system using leucine zipper proteins Jun and Fos
Two compatible plasmids pACYC184 and pBHR68 were modified to express and produce DBD-X and reporter LacZ, PhaP-Y and PHA granules, respectively. × and Y stand for a pair of tested interacting proteins (Figure 1C). To study the feasibility of this system, the leucine zipper domains of transcription factors Fos and Jun, which have been confirmed to have a strong interaction, were chosen as model interaction partners. The reporter gene lacZ was placed under the control of the phaP promoter. In principle, DBD-Jun fusion protein binds continuously to the phaP promoter region as soon as they are produced, repressing the expression of LacZ until PHB granules begin to form. The interaction of bJun and bFos will force DBD-bJun and bFos-PhaP to form a new complex, namely, DBD-bJun:bFos-PhaP, which is similar to a reconstituted 'PhaR' protein. As soon as PHB granules are formed, DBD-bJun:bFos-PhaP will be released from the phaP promoter region, resulting in LacZ expression. As expected, a strong expression of LacZ was observed (Figure 2, bar 9). This is the first evidence showing the feasibility of this two-hybrid system.
Confirmation of the LacZ expression only resulted from interacting proteins
In this bacterial two-hybrid system, there are several indispensable elements: phaP promoter, DBD, PhaP, lacZ reporter gene, interaction protein partners and PHB granules. Possible influences of interactions among these elements themselves would lead to false positive results, so experiments must be performed to confirm that the LacZ expression only derives from the interaction between protein pairs.
Firstly, we investigated whether LacZ could express normally under the control of phaP promoter. For this purpose, plasmid pOZ was constructed (Figure 2, bar 1 and Figure 3A), its expression resulted in a high β-galactosidase activity comparable with bJun and bFos interaction group (Figure 2, bar 9), confirming the effectiveness of phaP promoter to regulate LacZ expression.
Secondly, plasmid pDBD-Z was constructed to investigate whether DBD can inhibit LacZ expression when PHB granules are not available (Figure 2, bar 2 and Figure 3B). Only little β-galactosidase activity was observed when pDBD-Z was co-expressed compared with strong β-galactosidase activity observed when pOZ was expressed alone. Therefore, DBD can be considered to suppress LacZ expression.
To further exclude the possibility of LacZ expression by elements other than interaction protein partners, following studies were performed: To investigate whether DBD could interact with PHB granules, PhaP or bFos, their corresponding plasmid combinations (Figure 2, bar 3-5 and Figure 3C-E,) were transformed into E. coli XL1-Blue to produce these elements. If interaction happened, a false positive LacZ activity could be detected. Similarly, we detected whether PhaP could interact with bJun (Figure 2, bar 6 and Figure 3F), and whether the interaction between DBD-bJun and bFos-PhaP or bFos (Figure 2, bar 7,8 and Figure 3G,H) could also direct to LacZ expression when PHB granules were absent, in other words, whether PHB granules are essential for LacZ expression. Obviously, LacZ activities produced by all above groups of recombinants (Figure 2, bar 3-8) were on the similar low level as that detected as a background level shown by the wild type E. coli XL1-Blue containing no plasmid (Figure 2, bar 12). These results clearly demonstrated that the LacZ activity resulted only from bait and prey interaction.
Study of the two-hybrid system using different interaction protein pairs
To further prove the feasibility of this two-hybrid system, two more interacting protein pairs, namely, bJun:bJun and bJun:bATF2 were employed. Plasmids pJun-P-CAB and pATF2-P-CAB harboring genes of bJun-PhaP and bATF2-PhaP, respectively, were transformed together with compatible plasmid pDBD-Jun-Z into E. coli XL1-Blue strains for their LacZ activity studies. From Figure 2, bar 9-11, it became clear that interacting pairs of bJun:bFos, bJun:bJun and bJun:bATF2 all showed strong LacZ expression with different strength levels, the strength of interaction correlated well with the β-galactosidase activity: bJun:bFos>bJun:bJun>bJun:bATF2.
Feasibility study of this bacterial two-hybrid system
As a successful two-hybrid system for studying protein-protein interactions, the expression level of the reporter gene should directly reflect the interaction strength of proteins. In addition, background expression of the reporter gene should be as low as possible, because the low background noise will improve the accuracy of the method to study proteins with low interaction strengths.
To test the feasibility of our bacterial two-hybrid system, we chose three extensively studied model interaction partners, namely, bJun, bFos and bATF2. Previous multicolour bimolecular fluorescence complementation (BiFC) assays [15] showed that heterodimer bJun:bFos and homodimer bJun:bJun were able to coexist in living cells, and bJun:bFos had a 60% content in the cells compared with only 35% content of bJun:bJun, indicating that the interaction strength of bJun with bFos was stronger than that of bJun:bJun, the bJun:bATF2 being the weakest. That is to say that the strength order among these three pairs should be bJun:bFos>bJun:bJun>bJun:bATF2. This reported order was consistent with the β-galactosidase activity order of the three interacting protein pairs studied using our two-hybrid system (Figure 2, bar 9-11).
In principle, the background expression of the reporter gene may result from the undesired weak interaction between essential elements involved in this two-hybrid system, such as DBD and phaP promoter region, DBD and PhaP, DBD and PHB granules, PhaP and bait, and so on. In this study, several experiments to detect LacZ expression resulted from these undesired interactions were conducted (Figure 2). Results clearly revealed that β-galactosidase activity maintained at a very low level when PHB granules were not formed in the cells, while weak β-galactosidase activity could be detected when PHB granules existed in vivo although its activity was very low compared with that produced by the bJun:bFos group. The following reasons may explain this low β-galactosidase activity: Firstly, DBD used in this study consists of a part of predicted PHB binding motif and the full DNA binding motif predicted by Pfam software to retain a strong DNA binding ability. This part of predicted PHB binding motif included in the DBD used here may still have a low binding ability to PHB granules, thus, a very weak LacZ expression was initiated when PHB granules were produced. Secondly, at the very beginning, genes of DBD and LacZ were transcribed and translated simultaneously in cytoplasm of E. coli, the earliest expressed DBD could turn to anchor on promoter region of lacZ, repressing its subsequent transcription. However, previously transcribed LacZ mRNA can be translated normally, this part of LacZ may contribute to the small amount of β-galactosidase activity.
Merits and drawbacks
Bacteria based two-hybrid systems allow the rapid analysis due to their faster growth rate compared with that of yeast, greater permeability to small molecules, absence of a requirement for nuclear localization and the possibility of studying proteins that are toxic when expressed in yeast [23].
When compared with other bacteria based two-hybrid systems, our system has additional advantages: all essential elements including PHB synthesis operon phaCAB, DBD, PhaP and the phaP promoter region are extrinsic in E. coli, therefore, we need not perform complicated genome DNA manipulations in the host strain, and need not worry about the influence of intrinsic components employed by other bacterial two-hybrid system. This is helpful to avoid intrinsic component derived false positive results. In addition, all essential elements used in our system are harbored by two compatible plasmids. Thus, this two-hybrid system can be easily transferred to other prokaryotic strains to obtain a better environment for production of bioactive proteins.
Results presented here have proven the feasibility of this bacterial two-hybrid system. However, further research should be carried out to obtain an optimal DBD without any PHB binding ability, which will contribute to a much lower background expression of reporter gene. On the other hand, PHB granules are essential for this two-hybrid system, even though plasmids can be modified for expression in eukaryotic strains such as yeast, the low yield of PHB in eukaryotic strains still limits the application of this system to study proteins requiring post-translational modification. PHB production ability is different depending on strains, experiments should be conducted to study proper culture conditions for sufficient PHB granules accumulation when this system is to be transferred to another strains. Sufficient PHB granules accumulation means sufficient space to attach all expressed PhaP fusion. For example, when bJun and bFos were employed as an interaction pair for our study, a relationship between β-galactosidase activity and culture time was established, the time point when β-galactosidase activity began entering its stationary phase was considered as the proper culture time (data not shown).
A bacterial two-hybrid system based on two separate domains of PHA synthesis regulatory protein PhaRRe was established. DNA binding domain of PhaRRe fused with a bait protein was able to anchor on promoter region of lacZ reporter gene to repress its expression, PHB binding domain fused with a prey protein was found attached to PHB granules produced in vivo. The interaction of bait and prey forced two fusions to form a reconstituted "PhaR" which was dropped from the promoter region, releasing the expression of LacZ. LacZ expression level depended on strengths of bait and prey interaction. Results from three extensively studied model interaction proteins bJun, bFos and bATF2 confirmed the feasibility of our two-hybrid system.
The research was supported by Li K-Shing Foundation and National High Tech 863 Grant (Project No. 2010AA101607 and 2006AA020104), as well as the State Basic Science Foundation 973 (2007CB707804).
Authors' Affiliations
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Child Seat Inspections- Available through the Ocean County Sheriff's Department
- Firearms applications shall be available through the Records Department during regular business hours, Monday through Friday, 9:00 a.m. through 4:00 p.m.
- You Must appear in person to retrieve any documents as information must be verified, please bring your firearms ID card if applicable and driver's license
- Fingerprinting services shall be available by appointment during regular business hours, Monday through Friday, 8:30 a.m. through 4:30 p.m.
- Fingerprinting services shall be available Wednesday evenings, 4:30 p.m. through 7:00 p.m. by appointment only.
Additional evening appointment times may be available, at the discretion of the Chief of Police.
- The fee for the fingerprinting service for Township residents is $5.00/per fingerprint card and $7.50/ per fingerprint card for non-residents.
- No fee shall be charged for fingerprinting services when they are required for the issuance of licenses or permits to be issued by the Township.
Child Identification program is offered at various times throughout the year. Child ID can be done for your organization or group by contacting Sgt John Convery at 732.833.3015
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During 1994, we celebrated the 150th year of Jackson's existence as an incorporated municipality in the State of New Jersey. As the handwritten Legislative Act reads, "Having been three times read in the Council (the State Senate) on February 29, 1844,"


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This article is about the last press conference that Obama had, just a couple of days before the new president elect is inaugurated. In his last press conference Obama opens up to the reporters and answers any questions that they might have. They bring up a lot of questions that range from Donald Trump to LGBT rights. The response that Obama gave to one question though, really caught my attention. The question that a reporter asked was if he believed that there would be another black president. Obama answered by saying that he believed that there would be many different types of presidents, like a woman, Latino, Jewish, or Hindu.
This relates back to our class in the way that religion was used in order to address a question in the public sphere. Obama believed that there could be a president that was Hindu and not just a president that was Christian. He made it known that Christianity and Hinduism were of equal status. Obama did not create this hierarchy of religions like the scholars that we have been reading and learning about. It could be thought of as a way of saying that Hinduism and Christianity are both at the last stage of Tylor's unilinear model of Savagery – Barbarism – Modern. The religions that are practiced in the U.S. have all gone through this evolutionary model because they are all still modern and of equal status, due to Obama's belief in their being a president that practices a different religion.


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The Double 6 Designer Domino Cards are a unique hand crafted set of 2 x 3 inch Tile-Like cards that are the same as a Regular set of Dominoes from Double 0 to Double 6!
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Annual Dues
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Checks can be mailed to
Bayport Blue Point Heritage Association
PO Box 4
Bayport, NY 11705
The Bayport-Blue Point Heritage Association was founded in 1983 and continues to be an active organization with over 300 members. Our mission is to preserve Bayport and Blue Point's history by collecting and maintaining historical information on the towns, their people, and buildings.
Our hard work has restored Meadow Croft, the summer estate of John Ellis Roosevelt, into an historic museum. Tours are available on Saturdays and Sundays from June through October and upon request.
In addition to the Autumn House Tour, other events offered by Bayport-Blue Point Heritage Association include Christmas at Meadow Croft, a historical boat cruise, summer concerts, museum visits, and special exhibitions.
Monthly membership meetings are held the second Sunday of the month from September through June. Meetings are from 2-4 and feature speakers making presentations on local history. Refreshments are served, so come to the Bayport-Blue Point Library and join us. New members are always welcomed!
Please consider joining The Bayport-Blue Point Heritage Association! Join Now
Join us for Christmas music played on the grand piano, rooms beautifully decorated for Christmas, learn some of the history of this historic home, don't forget to get your raffle tickets to win one of the many baskets and decorations. Head on out to The Carriage House for St. Nicks workshop - visit with old St. Nick himself, have your picture taken, enjoy some cookies and cocoa and view the great train display! (Be sure to visit the main house first to get your pass to the Carriage House)
Come and take a private guided tour of Meadow Croft all aglow with Christmas! Refreshments will be served. This event is for adults only. Be sure to reserve your spot early, space is limited.


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Welcome to Village Voice, the section of Gaylife Manchester where we hand the reins over to one of the prominent figures within our community for a bit of a reminisce and an occasional good old rant.This month Gerrard Woods, General Manager of Cruz 101 casts his eye over the last 21 years of life in the UKs largest gay scene, and raises concerns about the potential EMRO hanging over the village…
In 1991 the area now known as 'the Gay Village' was little more than a collection of disused mills, run-down buildings and an undeveloped canal side; a red-light district to some, but the beginnings of the Gay Village to others and an opportunity to bring the gay community into the mainstream to us at Cruz 101.
We picked a partially derelict shipping warehouse built in 1864 as the place to show Manchester what the community was capable of and created a nightclub that surpassed even what we thought it could be.
It wasn't all plain sailing though. Even before the club had opened prejudice was rife with a straight refusal for a licence. 'Not sufficient need or interest' was the reason given for this denial but we were not to be denied. Fighting for the right to come out of the shadows and be proud of the community we sought to service, a legal battle followed ending up in the High Court with a victory for equality. Of sorts.
Having demonstrated the interest by signing up thousands of members we were not granted a full on-licence. Instead we were granted a private members licence which although disadvantaged us, allowed Manchester's first mainstream gay club to start trading. Since that day in May 1992 we have had over 100,000 members, over three million visitors and have had the honour of being the place where so many have come to meet their loved ones and some have brought their loved ones after they have passed away.
Once a red-light district, the Gay Village became, over time, one of the most fashionable areas to socialise in Manchester – the stigma of homosexuality being replaced with its fabulousness.
Over the past 21 years we've watched the Gay Village blossom into the vibrant community it has become today. Here at Cruz we have remained true to our original intentions; to create a safe gay space that is welcoming and affordable to the thousands of our friends who visit us each week. Many venues have opened and closed, and we have certainly seen styles change – in terms of music, fashion and venues. Once a red-light district, the Gay Village became, over time, one of the most fashionable areas to socialise in Manchester – the stigma of homosexuality being replaced with its fabulousness.
As strides towards equality were made we were involved in campaigning for equal age of consent and the abolition of the much hated Section 28, and much progress was made. Equality was the new watch-word and this included television. One of our customers, Russell T Davies, grasped this opportunity and wrote the hit series 'Queer as Folk'. Set in the Gay Village it told the not too-far-fetched story of life on the Manchester gay scene featuring a mix of characters portraying the diverse regulars who visited and found acceptance and safety within it. As I say, from my perspective it was not a million miles away from real life and I know many of our customers who could personally identify with it.
There was a problem though. With the celebrity of the programme came a wider awareness of the Gay Village and a far bigger desire by the wider community to 'see what it was all about'. The Gay Village became a destination for visitors keen to explore and find out if what they had seen on their screens was a true reflection of life on the scene; if the excitement, promiscuity and social life lived up the fame. Some thought it had turned gay bars into goldfish bowls, others thought it had given the gay community the exposure it needed to finally achieve true acceptance. Whatever your view, it certainly made a big change.
With the charge for equality had arrived a whole new dilemma for us – how to provide our safe gay space for our community without ourselves being discriminatory; how to integrate without losing our identity. It is a dilemma that continues to exist for the community today with some of the belief that there is still a need for the safety and security afforded by our little enclave and others who see it as an obstruction to true equality.
So where do we find ourselves today? This rollercoaster ride we've been on for the last 21 years is certainly not immune from the 'boom and bust' of recent times. Manchester had become the largest gay destination in the UK and in the years leading up to the middle of the first decade of the new millennium the Gay Village had grown bigger and bigger to service not only the gay community but also the more diverse crowds who had chosen to socialise in this most fashionable of districts. With the onset of recession came diminishing attendances. With cheap supermarket alcohol and shrinking budgets house parties became more fashionable. The gay community continues to support the Gay Village but it is certain that the recession has been diffi cult for everyone.
And now, as the business owners dig deep to keep their businesses afl oat to service our community another spectre has appeared on the horizon. Reports of our Council's intention to close our community down after 3am cast a long shadow over the Gay Village.
This is an area which has, for the twenty one years we've been here, been a safe space that has facilitated the social lives of the gay community. Partying until the early hours of the morning has been one of our community's privileges and we have excelled at it. In a safe way. Now it's the intention of Manchester City Council to enforce a terminal hour of 3am on all of the venues who operate within the Gay Village using new legislation called an EMRO. Why? By their own admission crime is falling month on month, year on year. Their reason is that the numbers of crimes that occur on the streets of the Gay Village is disproportionately high.
They say that the crimes are not committed by the venues or their visitors; that they are in fact committed by people from other areas of the city gravitating towards the Gay Village as it has later licensing hours, many food outlets and several taxi ranks. So is this the spectre of homophobia?
I hope not but when you look at the facts – our community has been descended upon by people who cause trouble; our bars and clubs operate responsibly and our customers behave when socialising yet the area defined as the Gay Village is to be the subject of an EMRO, it's hard to see how it isn't.
Surely when looking at the bigger picture the answer is not to close down the Gay Village but to deal with the people causing the trouble? Would a shopping centre be closed down following a spate of shoplifting? No. The police would be required to deal with the offenders. Targeting an area which is defi ned by its community has to be wrong.
There's obviously going to be a passionate debate over the coming weeks and months about the merits and impacts of what an EMRO might do to, and what the future holds, for the Gay Village.
We've come a long way in the last twenty one years and I hope that sense is seen and that future is a bright one.
Gerrard Woods
General Manager, Cruz 101


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EO Austin is chartering a flight and renting a house/yard across the street from the track to act as a launching pad and landing spot before and after the Kentucky Derby. You won't want to miss this event!
Where: Kentucky Derby
700 Central Ave
Louisville, KY
Key Details:
- Price includes charter flight, buses, pre and post party and an infield race ticket
- Brian Peters will be helping members secure upgraded tickets. All levels are available. Group seating available to sit with your forum or other EO members.
8am: Charter Flight – Plane departs from Austin
11am: Arrival and bus pickup
11:30am: EO pre-party with mimosas and mint juleps, light appetizers.
1:00pm – 6:00pm: Kentucky Derby – RACE DAY!
6:00pm – 9:00pm: After party with food, cocktails and a SUPER FUN band!
9:30pm: Bus Pick-up
10:30pm: Plane departs from Louisville
12:30am – 1am: Plane lands in Austin
To learn more, contact Emma: email@example.com


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Ralph Ellam is the g-g-g Grandfather of my husband, Mark. When I first began researching the Ellam family many years ago I had very little to go on. Mark's grandfather, John "Jack" Ellam, Jr. walked away from the family when Mark's father was young and very little was known about his ancestors.
I have since been able to trace the family roots back to St. Helens, Lancashire, England, where I discovered Ralph living in 1850. I found Ralph's marriage record in 1853 to Elizabeth Greenall and sadly discovered his death in 1859 at the age of 35 due to an accident in the coal mine he was working in. At the time of his death his son, William, (my husband's g-g grandfather) was only 9 months old.
With that I closed the case on Ralph. Afterall, I had managed to find his birth date in England and Wales Christening Records , 1530-1906, his marriage record to Elizabeth in the England and Wales, Free MBD Marriage Index: 1837-1915 and his death in the England and Wales FreeBMD Death Index 1837-1915. I sent for, and received, copies of his marriage and death certificates. I was even able to find his burial location in St. Helens Cemetery through the St. Helens Council - Deceased Search.
I thought I had learned all I could about Ralph's relatively short life. Until this week.
Ancestry.com just put online the England & Wales National Probate Calendar (Index of Wills and Administrations), 1861 - 1941. I plugged in the Ellam name to see what I could find and was shocked when the first result turned out to be Ralph! 18 years after Ralph died, Elizabeth his widow, was granted the effects of Ralph's estate.
30 April 1877. Administration of the effects of Ralph Ellam late of Parr near St. Helen's in the county of Lancaster Miner who died 12 April 1859 at Parr was granted at Liverpool to Elizabeth Heaton (Wife of Lambert Heaton) (formerly Ellam widow) of St. Helen's the Relict.
Now to visit the Probate Registry's Website and order a copy of this probate and determine why it took 18 years to close an estate worth less the $100.00 USD.
What have I learned from the case of Ralph Ellam? Leave no stone unturned. I assumed since Ralph was unable to read or even write his own name, and that because he worked in the coal mines, he would have left no estate behind. I was wrong.
Another page in the story of Ralph's live has been turned. I can't wait to read this chapter.......


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The 3D Printing & Additive Manufacturing Industrial Applications Summit 2013 (London, 19-20 November) is the world's first industry-driven AM forum designed specifically to identify the real business benefits and new opportunities these technologies present. Bringing together a cross-section of industry leaders and experts, participants will be able to gain first hand insights on successful commercial applications, state-of-the-art machine and material developments and explore solutions for addressing the complex intellectual property issues arising from the new manufacturing processes.
They will hear from BAE Systems, GE, Renault, Siemens, Bayer MaterialScience, Nokia, Dyson and other users of additive manufacturing technologies as they explain the strategic reasoning behind investment and their visions of what these new processes really mean in terms of commercial opportunities for industrial production.
As a subscriber of IEN Europe, you will get a 15% discount. All you have to do is quote 'IEN15' at the checkout here
To view full speaker line-up and agenda go to www.3d-printing-additive-manufacturing.com/


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Well the other day, I went to my friend Lauren's party! The theme was 'My Kitchen Rules' cooking party. (My Kitchen Rules is an Australian cooking TV show where teams from different states cook against each other and the clock!) So all the girls were divided onto teams from different states, and we had half an hour to cook a sweet (there was recipes to choose from) and Lauren's parents were the judges and chose who won! It was a lot of fun! My close friend Cassie and I were on the South Australian team and we cooked a lemon curd tart and won!! This is the second cooking party I've been too, and probably won't be my last : )
(to view the other party, go here.)
|doesn't it look delicious?!
|The dish that Lauren and her partner made. Fancy chocolate-Anzac slice!
|all of us girls just before the birthday cake
|the table after us girls had feasted on all the sweets
|Lauren's beautiful birthday cake!
|Lauren and I in our pretty aprons!
Have you ever been to a cooking party before??
p.s. if you want to view the recipe and more photos of the lemon curd tart, I will be posting about it as soon as I can on Life is Sweet!
p.p.s. Guest posted over here! Please read it and tell me what you think!
p.p.p.s. One more thing. Please read Sponsors... post if you haven't already!
p.p.p.p.s. I promise this won't happen all the time! There's just something else I have to tell you Followers! Lucia from Umbrella Girl is hosting a giveaway where she's giving away a whole blog design! The whole lot! Remember to go over and enter if you haven't already : ) Giveaway ends on the 23rd.


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Toni Erdmann Is the Delightful Christmas Movie You Might Not Want to See With Your Dad
When you head out to see a Christmas movie this holiday weekend, there's a good chance you'll opt for Damien Chazelle's La La Land, Mike Mills's 20th Century Women, Martin Scorsese's Silence, or Denzel Washington's Fences. (If you've got your eye on Office Christmas Party or Collateral Beauty, this story may not be for you.)
But why not make the unlikely choice and buy a ticket for a nearly three-hour-long German-language film about international management consulting set in Romania instead?
That would be the writer/director Maren Ade's Toni Erdmann, one of the weirdest, funniest, most unexpectedly delightful movies of the year. If you're skeptical, check out Vogue.com's film critic John Powers's holiday movie report card, Rotten Tomatoes, or A. O. Scott, who wrote in The New York Times, "if a single movie were enough to silence reports of the death of cinema, it would be this one."
The title refers to the prankster alter ego of Ade's main character, Winfried Conradi (Peter Simonischek), a divorced, middle-aged German music teacher. We're introduced to Winfried mid-joke: A mailman rings his doorbell to deliver a package. "Toni!" Winfried calls into his own house, informing the postman that his brother was just released from prison after serving time for sending mail bombs. Winfried disappears, only to reemerge seconds later, wearing a set of gnarled fake teeth, barely in disguise. "Thanks," he deadpans to the mailman, as though meeting him for the first time. "Now I'm really looking forward to defusing it."
You'll see a lot of those fake teeth in the 162-minute span of Toni Erdmann. As you watch Winfried move through the world—clowning at the school where he teaches, pulling pranks on annoyed family members, in the presence of disbelieving strangers—it will begin to occur to you that his penchant for practical joking borders on the compulsive.
Never is that more apparent than when he uses his gags to try to connect with his only child, his grown daughter Ines (Sandra Hüller), a harried, type-A consultant who benignly neglects her relationship with her father while working around the clock in Bucharest to help her oil-magnate client justify a massive round of lay-offs.
When Winfried arrives unannounced in Romania to surprise Ines with her birthday present—a cheese grater and some cash—and then, to her dismay, long overstays his welcome, it exposes their epic clash of world views and lifestyles: alpha vs. beta; corporate vs. hippie; continental vs. not. Winfried's presence also highlights a major difference between how men and women move through the world: Ines, a stressed-out striver, takes herself too seriously, but she does so because otherwise no one else would. Winfried can poke fun—at himself, at others, at basically everything—but that's in part because he's automatically granted entry to circles that his daughter must fight to access.
There's a strong critique of capitalism and globalism that runs through Toni Erdmann, and a feminist edge as well. But neither strikes at the heart of the film, which is really about Winfried and Ines's idiosyncratic relationship. Ade paints father/daughter tensions with an empathetic, nuanced brush, revealing her two main characters as two sides of the same coin. There's some of Ines's corporate-ladder-climbing doggedness in Winfried's tone-deaf refusal to leave his daughter be. There's something of Winfried's absurdist sense of humor in Ines, too: We see it best in one of the stranger sex scenes I've ever witnessed on the big screen. Ines shows up for a hotel room assignation with a colleague, then, bored by the cliché of it all, demands that he jerk off onto a tea cake.
And we see it again during the film's climactic—not that way—episode, which I won't spoil except to say that it involves a naked brunch party and a very surreal appearance from Winfried decked out in a Wookiee-like Bulgarian Kukeri suit.
Your choice to go see this movie on Christmas should probably be directly correlated to your comfort sitting through those two scenes in the company of your relatives. But it's that very earthiness that makes Toni Erdmann so unique, so unexpected, so refreshing, so humanist. Winfried's joking, we begin to see, is both his way of connecting with people and of keeping them at arm's length. For Ines, the rigid choreography of the business world serves the same function.
Both father and daughter are, in their way, performers: he with his teeth and wigs and disguises; she with her power suits, her chignons, her high heels.
The only trouble is that they see through one another's act. And that, after all, is the rub with family.