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During the time leading up to the Revolutionary War the American people were very restricted in many areas of how they lived and how they did business. Before the Revolutionary War it was illegal to print a Bible in the Colonies. All Bibles had to be shipped in from England. The reason for this was the King did not want a version of the Geneva Bible printed in the Colonies. That Bible did not hold the king in the position that he wanted to be in and that was supreme ruler, almost a god. It was a situation where the king decided what you needed to know concerning the Christian religion. The importance the Geneva Bible to the Pilgrims and the Founders can be explained.
To understand it here is a bit of history of the Geneva Bible: “It is important that we spend a little time looking at why America was such a desired place to go during this time. America had begun to establish a society that was freer than any other nation in the world. That society was being built on the teachings of the new Geneva Bible. Up until the Geneva Bible no Bible was available in the English language nor were there any commentaries to explain many aspects of the teachings found in scripture. The Bible had just begun to be translated into the English language. The New Testament was first printed in 1526 by William Tyndale. In the process of translating it he was forced to flee England because of persecution from the Crown. He fled to Germany and then to Belgium before finishing the translation. Six thousand copies were smuggled into England and that caused Tyndale to be hunted down and imprisoned. Then on March 6, 1536 he was strangled and burned at the stake, all for printing the New Testament in the English language.
It was in 1553 during the reign of ‘Bloody Mary’ Tudor when the English were pushed to return to Roman Catholicism. She ordered the burning of all copies of the Bibles in English and caused more than 300 reformers, pastors and Bible translators to be burned at the stake. This caused about 800 English scholars to leave England, for safety reasons, and they gathered in Geneva culminating in the grouping of the finest theologians and Biblical scholars in the world at that time. There between 1557 and 1560 they began to translate the entire Bible into the English language. In the Geneva Bible were footnotes and commentaries from John Calvin, Thomas Sternhold, John Hopkins, William Whittingham, Miles Coverdale, John Knox and many more. These commentaries infuriated King James because they began to teach against a monarchy and totalitarian rule, supported individual freedoms, property ownership and many other things that took power and authority away from the king so in 1606 he ordered that a Bible without notes and commentaries be made available for him. This became the King James Version we have today.
The Geneva Bible was the first Bible to be completely translated to English from the original language.” (Emphasis mine throughout) These commentaries talked about having men who feared God in places of authority, owning property, and individual freedoms. From this we can see that it was this Bible that began to build the fire for liberty in the hearts and minds of not just the Pilgrims but kept that fire burning all the way to the Founders and beyond. It was said that that the key liberties that were in the Declaration of Independence had been preached from the pulpits of America since the early 1600’s.
When America was beginning to be established the Pilgrims began to experience liberties that were unknown to the common person in the world at that time. One could pursue the vocation that he was interested in. In Europe there was still a strong cast system and what your father did you did whether you wanted to or not. This freedom was built the foundation for the greatest free market system the world had ever seen.
One could worship as one believed was the manner in which he believed best. In England the Church of England was the prominent church. You paid taxes that supported the church and the government dictated the doctrines of the church. In America you could worship freely and the pastors were free to preach the uncompromised Word of God. This exposed the inhabitants of America to a way of life that was not known anywhere else in the world.
By Pastor Roger Anghis – NewsWithViews.com –

domain: library


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View Full Version : First ever minis
01-16-2005, 10:51 AM
I just started painting some minis because I bought a pack of 2.1/4 x 3.1/2 inch frames in a cheap shop the other day.
They are all done in acrylics on watercolour board.
Any comments or crits would be welcome.
01-16-2005, 12:33 PM
Very nice. I really like the one with falling snow.
What's a "cheap shop"?
01-16-2005, 01:44 PM
Aren't they fun!
I can tell by your work that you will easily get hooked! *grin*
Welcome to the world of miniatures.
01-16-2005, 04:49 PM
These are all Excellent. I love them all. :clap: :clap: :clap: :clap:
The snow is very effective, and the sunset really glows -- there is something in each one that says to me, "Frame me, and put me in your home" :) :)
Hope to see more from you.
01-16-2005, 05:12 PM
These are beautiful minis!!! Love that falling snow and sunset!! Looks like you're a natural at miniatures!! :clap:
01-16-2005, 05:54 PM
These are very nice - each one worth framing. What do they look like in their frames?
01-16-2005, 08:18 PM
Wonderful style. I'd like to see them in their frames too. :D
01-16-2005, 08:25 PM
Nice job on these :clap: ! You're a natural mini :D !
01-17-2005, 04:34 AM
Thank you everyone - I'm thrilled with the way these turned out.
I will take some pics of them framed - but I am not sure how the digicam will deal with the glass in the frames.
Lady Rando - we have shops here in the UK that started off selling everything for £1, but these days they don't stick to the £1 so tightly and they sell sell pretty much everything from toothpaste to rugs at very cheap prices. It's pot luck what you get in the shops as they change stock regularly. I got the frames for £1.49 for four.
01-17-2005, 05:30 AM
Hi Cryla...those are called Dollar stores here...lol
I like your style, really like the sunset. Can't wait to see them framed. I'm going tomorrow to search for frames for some of mine.
01-17-2005, 12:25 PM
Oh yes please show us them framed. Leave the glass out when you do it and they will look great!
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domain: hobby


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One of the common Event prizes is an Event Trophy. Upon initial Camp arrival if there's free space around them Trophies will typically spawn 4 Event Eggs of a type that can be purchased in Nest form in the Egg Shop for Gems. They will then become a dispenser for Trophy Chests.
Trophies of this type can be sold for 2,000 Coins.
For Trophies with alternative behavior, please see below.
Right after unbubbling the Event Trophy, and after that, every 10 hours spent in the Camp it can be tapped for a level 1 Trophy Chest. Level 1 Trophy Chests will typically give either a Young Dragon Tree or a Wondrous Life Orb upon tapping. If you merge them to level 2 they will give either a Young Dragon Tree, a Great Life Orb, or rarely (usually 10% chance) an Event Egg corresponding to the Trophy's Event origin. These Event Eggs can usually be purchased individually in the Egg Shop for Gems.
Event Trophies and Trophy Chests
Event Trophies with alternative behavior
These Trophies either are pure Camp decorations that do nothing, don't spawn 4 Event Eggs or can only be tapped for things different from the Trophy Chests. For more information please see their respective pages.
- Baby Moo
- Entwined Passion Trees
- Garden Gnome
- Giant Lucky Coin
- Haunted Mansion
- Red Rose
- Snuggles McCuddlesworth
- The Great Pumpkin
- Winter Wonderland Snow Globe

domain: game


text:
Press Release Summary:
Suitable for rotating, oscillating, and linear movements, igubalÃ‚Â® K-Series rod end bearings are available in clip version with different ball materials: 6 high-performance plastic blends as well as stainless steel. Balls come in iglideÃ‚Â® L280 for applications with high wear rates, cost-effective iglide R, low-friction iglide J, iglide J4 with low moisture absorption, chemical- and high temperature-resistant iglide T500, and iglide UW for underwater applications.
Original Press Release:
Rod End Bearings with Different Ball Materials
igusÂ® now offers its igubalÂ® K-Series rod end bearings in a clip version with different ball materials. The new clip-in feature makes the dimensionally interchangeable rod ends more versatile. The balls are available in six high-performance plastic blends, as well as stainless steel.
Each iglideÂ® material has a unique set of properties that help determine its suitability for a given application. Currently, the spherical balls come standard in the material iglide L280, which is ideal for applications with high wear rates.
The balls are also offered in iglide R: a cost-effective material, iglide J: a low-friction material, iglide J4: a material with low moisture absorption, iglide T500 for environments exposed to chemicals and high temperatures, and iglide UW specifically for underwater applications. All five iglide materials are self-lubricating, corrosion resistant and lightweight.
igubal rod ends are suitable for rotating, oscillating and linear movements.
To request a free sample, visit www.igus.com/quickorder or call 1-800-521-2747.

domain: chemistry


text:
Pope vows war on abuse; survivors say let down
Pope Francis has promised zero tolerance on sex abuse at the end of a landmark conference. But survivors and activists say without concrete action such as defrocking abusing bishops, they..
Credit: Rumble Duration: 01:50 Published 3 weeks ago
Church must fix 'systematic failures' on abuse
At the second day of a summit on sex abuse, leading cardinals have said the Catholic Church must repair the "systematic failures" that enabled sexual abuse to take root around the world, and bishops..
Credit: Reuters Studio Duration: 01:39 Published 3 weeks ago
Sex Abuse Survivors Demanding Accountability From Vatican
In advance of a four-day summit on clergy sex abuse at the Vatican, survivors say they want church officials to stop claiming ignorance about abuse that's gone on for decades.
Credit: CBS 2 New York Duration: 00:47 Published 3 weeks ago
Vatican Meets With Abuse Survivors
According to CNN, Organizers of a key Vatican summit on clergy sexual abuse met with abuse survivors from around the world on Wednesday.
Afterward organizers said that the meeting will help them..
Credit: Wochit Duration: 00:44 Published 3 weeks ago

domain: sociology


text:
Over their drinks, theater people sometimes play a game: they dream up casts for great plays. With opium-pipe prodigality, they sometimes devise a Hamlet in which Lionel Barrymore plays the second gravedigger, a Macbeth in which Tallulah Bankhead plays the third witch. But they know that not only would the cost of such productions be staggering, but collecting all the right people would be a super human feat.
Not for nothing is Katharine Cornell the top-ranking actress in the U.S. theater as well as a successful producer as well as the wife...
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domain: artistic


Lady Bird Johnson was barely cold in her coffin before federal highway administrators began gutting her heralded beautification efforts.
Back in the 1960s, the former first lady fought to remove junkyards and billboards from the edges of American highways. Her theory: The nation’s landscapes reflect the human spirit, and beauty creates harmony while clutter creates anxiety. She and hubby Lyndon B. Johnson went on the warpath, but billboard lobbyists are a powerful bunch. Regulation proved difficult, but a watered-down bill was eventually passed. Now it’s even thinner stuff, since the Federal Highway Administration recently decided that digital billboards don’t violate the law’s prohibition against intermittent, flashing, or moving lights – strange, since digital billboards include all of those things.
A local fellow who goes by the internet name StreetSpamShark (he wants anonymity because he fears repercussion from pro-billboard folks) said digital billboards and their huge video screens will distract motorists. “Drivers are going to get into accidents, and people are going to get killed,” he said. A federal safety study on that topic is under way but won’t be finished until 2009. Critics such as the nonprofit beautification group Scenic America want a moratorium on digital signs until the study is completed and have accused the feds of caving in – again – to the billboard industry.
Praying to St. Caffeine
Candidate forums are usually simple, not to say boring, affairs. Potential elected officials tell various half-truths embodying what they think voters want to hear about who they are and what they’ll do in office. But even the politically jaded Static was dumbfounded by the performance of James Schull at a recent forum for those running for the District 97 seat in the Texas House.


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California is on the verge of becoming the first US state to require climate disclosures for large companies after its State Assembly approved the Climate Corporate Accountability Act (SB 253) yesterday (11 September).
If signed into law, the legislation will require all business operating in California that make over $1bn a year, roughly 5300 companies, to annually report their direct (Scope 1 and 2) emissions from 2026 and annually report their indirect (Scope 3) emissions from 2027. Notably the rule would require independent verification for these disclosures.
The bill's sponsor, Senator Scott Wiener, said: "These commonsense disclosures are already the norm in many markets - we need to standardize them to ensure investors, regulators, and consumers have the information they need to engage with major corporations in a warming world."
The bill was approved by the lower house with 41 votes in favour and 20 votes against. It now needs to be rubber stamped by a final Senate vote before Governor Gavin Newsom can sign it into law, which is expected to happen in the coming days. The California Air Resources Board will then have to approve regulations by 2025 to implement the climate disclosure requirements.
The previous iteration of the law didn't pass the Assembly last year as a significant number of moderate Democrats voted against it.
Wiener has built a wider political coalition this time around and made amendments to the bill to alleviate concerns over reporting indirect carbon emissions by weakening the repercussions for inaccuracies in Scope 3 reporting.
Momentum has been building behind the legislation for some months, with endorsements from lawmakers, climate advocacy groups and companies, all of which showed their support at a rally in Sacramento at the end of August.
This momentum has continued, the California Appropriations Committee approved SB 253 last week and Apple and Google both threw their support behind the bill in last couple of days.
SB 253 is co-sponsored by a range of environmental groups, including Ceres, the Greenlining Institute, Carbon Accountable, Sunrise Bay Area, and California Environmental Voters (EnviroVoters), who pushed back on an anti-disclosure lobbying campaign headed by the California Chamber of Commerce.
"Up against the most powerful and resourced interests in our state, the Assembly stood strong and advanced world-changing policy, yet again proving our state's critical role in the global fight against the climate crisis," EnviroVoters CEO Mary Creasman said in a statement. "As the 4th largest economy in the world California will move the world forward on climate action and accountability."
Meanwhile, the other side of the climate disclosure package, the Climate-Related Financial Risk Act (SB 261), awaits a vote on the Assembly floor. SB 261 would require businesses operating in California with over $500m in annual revenue to annually release TCFD-aligned disclosures on their climate-related financial risks.
The US Securities and Exchange Commission is expected to finalise and release its proposed climate disclosure rule in the coming months, although significant problems and uncertainties remain. Many hope that the California legislation will lead the way to nation-wide climate disclosure requirements, with CDP announcing that the "world is watching" before yesterday's vote.
"The world is looking to the United States to provide climate leadership, particularly as we strive to set measurement and disclosure standards," former IOSCO Executive Committee chair Jane Diplock said. "That's why all eyes right now are on California. The current Californian climate accountability bill (SB-253) would be a great step forward."

domain: politics


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Dr. Julie Sutcliffe Named a 2018 UC Davis Health Deans' Award for Excellence Honoree
The UC Davis Biomedical Engineering Department congratulates Dr. Julie Sutcliffe on being named a 2018 UC Davis Health Deans' Award for Excellence recipient.
Recognized specifically for her "outstanding contributions as a transformative leader in the area of Research," Sutcliffe will join other Deans' Award honorees at a formal celebration April 29.
Sutcliffe is a faculty member of both the UC Davis Biomedical Engineering Department and the UC Davis School of Medicine, as well as co-director of the UC Davis Biomedical Engineering Department's Center for Molecular and Genomic Imaging. Her research focuses on targeted molecular imaging agents for preclinical and clinical applications with a focus in oncology.
The Deans' Award for Excellence recognizes outstanding faculty contributions in the core mission areas of Research, Education, Clinical Care and Community Engagement. The purpose of the award is to reward the outstanding performance of a faculty member (Academic Federation or Academic Senate) and to acknowledge their contribution as a transformative leader. The nominee’s dedication to one of the mission areas is a focused and compelling example that exemplifies and aligns the UC Davis Health community with its mission of discovering and sharing knowledge to advance health. The nominee’s efforts must clearly contribute to UC Davis Health being a destination of choice to all those served.

domain: medical


The 832th session of the Supreme Council of the Cultural Revolution was held on Wednesday, chaired by Dr. Hassan Rouhani.
At the end of the meeting, Prof. Saeid Reza Ameli, Secretary of the Supreme Council of the Cultural Revolution, stated about the approvals of this meeting and said:" The main agenda of this meeting was the policies and action plans of the National Educational Research and Training Platform. They agreed and discussed the need to focus education and research services and pay attention to its cultural dimensions in cyberspace, especially in the context of Corona, where much of the country's education and research has been transferred to cyberspace."
"In addition to the corona conditions, which have required the use of cyberspace and electronic education and research in order to observe the physical distance, cyberspace is a new superpower that can provide the best quality in education and research under normal circumstances." Ameli said.
Referring to the logic of cyberspace that was discussed at the meeting, the secretary of the Supreme Council of the Cultural Revolution said: "Cyberspace is different from physical space, and in this space, when we create a capacity, this capacity can be used nationally and globally. Unlike physical space, where the classroom is formed in a physical space limited to the environment, in cyberspace, facilities have a transcendental dimension."
"In the first passion, discussions were held on how to create convergence and synergy in the platform of the unit (platform) in order to maintain and flourish creativity and innovation while using the capacities of the public sector, private sector and global capacities." Prof. Ameli said.
"This issue has three levels of infrastructure and technology, content and services, the infrastructure of which must be provided in the context of the national information network, and the levels of services and content is a distributed matter for which all national capacities must be used." He added.
The Secretary of the Supreme Council of the Cultural Revolution stated that the meeting emphasized that the creation of these capacities should not limit the activities of universities, ministries and agencies active in this field, adding that this capacity should be a central capability for synergy. For example, the presence of 700 systems of the Ministry of Health in this national platform and organizing them in terms of services will create a new capacity.
Prof. Ameli said: The members of the Supreme Council emphasized in this meeting that facilitating the provision of services and meeting the needs of the widespread users in the field of education and research, which is a large population of 20 million at the student level, as well as talking about a large scientific community of professors Is to be considered.
"Also, paying attention to religious and divine values in the virtual cultural space was one of the issues that were emphasized." He added.
Prof. Saeid Reza Ameli also said: "At the end of the meeting, the esteemed chairman of the Supreme Council spoke with the members about the need to formulate macro policies and be accountable for the field of education and research and pay attention to educational dimensions. These policies and action plans were approved by the Supreme Council."
It was decided that the Secretariat will hold meetings with the stakeholders of this project, such as the Ministry of Science, the Ministry of Health, the Ministry of Education and the Deputy Minister of Science and Technology, in order to redesign; The macro policies of this field should be reviewed and approved by the Supreme Council of the Cultural Revolution,he said.
the actions in this field will be in the hands of the public, private and other stakeholders and the creation of new and additional organizations that hinder direct activities will be avoided as one of the concerns of the members of the Supreme Council, Prof. Ameli emphasized.


Indoor and outdoor air has between 400ppm and 2,000ppm CO2 by volume. This means that for projects from measuring indoor air quality to measuring atmospheric CO2, a 0-1% (0-10,000ppm) CO2 sensor is your best solution. For applications like CO2 alarms in restaurants, breweries, or around stored CO2 tanks, a 5% CO2 sensor is used.
The SCD-30 is an NDIR sensor, which is a 'true' CO2 sensor that will tell you the CO2 PPM (parts-per-million) composition of ambient air. Unlike the SGP30, this sensor isn't approximating it from VOC gas concentration - it really is measuring the CO2 concentration! That means it's a lot bigger and more expensive, but it is the real thing.
Nov 11, 2020· These can optimize the air quality in rooms for more healthy and productive indoor living and working conditions. Figure 2: The increase of the concentration of CO2 (left axis) and aerosols (right axis) in a classroom over the duration of two lessons with a break. Figure 3: The XENSIV™ PAS CO2 sensor measures only × 14 × mm.
A carbon dioxide sensor or CO2 sensor is an instrument for the measurement of carbon dioxide gas. The most common principles for CO 2 sensors are infrared gas sensors ( NDIR) and chemical gas sensors. Measuring carbon dioxide is important in monitoring indoor air quality, the function of the lungs in the form of a capnograph device, and many ...
MH-Z14A is an Infrared Carbon Dioxide Sensor for accurate NDIR (nondispersive infrared) Co2 measurement using Arduino, STM32, Raspberry Pi. This NDIR Co2 sensor is a small size, common type sensor, NDIR principle to sense the presence of CO2 gas in the air. This NDIR Co2 sensor provides good selectivity, long life and non-oxygen dependent.
UNIVERSITE MOHAMED V FACULTE DES SCIENCES RABAT Projet de fin d’études Filière : La licence fondamentale en Informatique, Electronique et Automatique « IEA » Département de physique Réalisation d’une maison intelligente à base d’Arduino Réalisé par : EL YAHIAOUI Khadija BOUKOUTAYA Abdel Adem Encadré par : Le président des encadrants Mr. OUADOU Mohammed …
Adafruit Industries, Unique & fun DIY electronics and kits IR (Infrared) Receiver Sensor [TSOP38238] : ID 157 - IR sensor tuned to 38KHz, perfect for receiving commands from a TV remote control. Runs at 3V to 5V so it's great for any use, connect pin 3 (all the way to the right) to 5V power, pin 2 (middle) to ground and listen on pin 1.
CO2 Monitor, Air Quality Monitor, Air Quality Meter Monitor manufacturer / supplier in China, offering Carbon Dioxide Meter (CO2) Monitor Indoor Air Quality Temperature Humidity Ndir Sensor Amazon Hotsale SA1200p, Rechargeable Battery Powered Desktop Air Quality CO2 Monitor for Classroom, Portable Combustible Gas Detecting Alarm with Rechargeable Battery Powered and so on.
Crosman CO2 Powerlet Cartridges are designed to provide a snug fit and solid seal for reliable and consistent performance with almost any CO2 gas-powered air rifle or air pistol. Available in pack sizes of 5-count to 500-count, ideal for those just entering into the air rifle/pistol sport, seasoned/competitive target shooters and air gun clubs.
DFRobot released its latest wide range infrared CO2 sensor, whose effective range is up to 0~50000ppm. This sensor is based on non-dispersive infrared (NDIR) technology and has good selectivity and oxygen-free dependency. It integrates temperature compensation and support UART communication mode. Most importantly, the product is easy to use and ...
Telaire T6713 CO2 Sensor Module is ideal for applications where CO2 levels need to be measured and controlled for indoor air quality and energy saving applications, such as demand control ventilation. All units are factory calibrated to measure CO2 concentration levels up to 5000 ppm. Telaire T6703 Series | Miniature CO2 Sensor Modules.


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20 Minute Stir Fried Honey Ginger Sesame Noodles
20 Minute Stir Fried Honey Ginger Sesame Noodles...best for nights when you’re craving a saucy, salty, noodle, but want homemade...and quick!
1. Preheat the oven to 400° F. Line a baking sheet with parchment. Spread the cashews out on the baking sheet and bake 5 minutes. Add the sriracha, 1 tablespoon soy sauce, 2 teaspoons honey, and the sesame seeds. Toss to combine. Bake another 5 minutes, until toasted.2. Meanwhile, cook the noodles according to package directions, drain. 3. To make the sauce. In a jar, whisk together 1/2 cup soy sauce. 3 tablespoons honey, the rice vinegar, peanut butter, molasses, and a big pinch of black pepper. 4. Heat the sesame oil in a large skillet over medium-high heat. Add the vegetables and stir-fry until softened, about 5 minutes. Stir in the ginger, cook another minute, then pour in the sauce and bring to a boil over medium-high heat. Stir in the noodles and chili garlic oil. Toss to combine, cooking another 3-5 minutes until the sauce coats the noodles. 5. Serve the noodles topped with cashews and green onions.

domain: food


TU-Automotive Detroit: Connectivity at the heart of automotive innovation
We attended TU-Automotive Detroit 2019 and discussed how connectivity is driving new services for the cars of the future. Here is what we learned from the panels and presentations.
Ericsson has been making the conference rounds and in addition to this year’s Future of the Car event held in London, we recently attended the 2019 TU-Automotive show in Detroit. We’ve been seeing a developing trend - the automotive industry is rapidly headed towards connected, autonomous and electrified vehicles. At the heart of all these advancements will be the need to collect and process massive amounts of data while on the go. Making that all possible is wireless connectivity. Here is what we saw at TU-Automotive Detroit.
Harmony in the ecosystem
The automotive industry is changing its business models in order to capitalize on data, and more importantly, how to increase revenue streams. The connected technologies being introduced into vehicles are all part of a larger ecosystem and will need to be interoperable and able to work within it. As no single player can handle it all, we saw automotive OEMs express a great need to cooperate and co-invent to support the larger ecosystem. Also discussed was how all the collected data and intelligence will be shared, going forward, necessitating an open ecosystem.
Intelligent connectivity is helping the drive to Industry 4.0
Other trends were focused on the customer experience. Speech recognition and artificial intelligence (AI) have made it possible to seamlessly introduce the same personal assistant experience to which we’ve grown accustomed to using at home, in the car. AI is also the motor powering autonomous vehicles.
Ericsson’s VP of Global Sales Connected Vehicles, Juergen Daunis, participated in a panel on “Intelligent Connectivity – The fusion between 5G, IoT and AI,” along with IBM, Orange and Verizon. The panel highlighted how these technologies will all play critical roles in the car of the future and how when combined, will transform the auto industry into a digital services business. AI is also enabling OMEs to reduce warranty and recall costs, as well as create new business models by collecting and analyzing car sensor data to learn more about driver habits.
A lot of new technology has recently been introduced and this trend will not be letting up anytime soon. Unfortunately, current cellular networks are simply not technically capable of supporting the advancements of AI and the high speed, low latency needs of autonomous vehicles. When vital decisions need to be made by a moving vehicle, there is simply no time to wait for data to transfer. Aware of this and the current limitations, many regions around the world where autonomous vehicles are starting to be tested, require a human to always be at the ready to take over control, if the need arises. Edge technology will certainly help the computing required and lower latencies, but only 5G has the necessary capacity to make truly self-driving cars a reality from a connectivity standpoint.
Ensuring a smoother ride
One final thing that rang clear at TU-Automotive was that we’ve only just begun to scratch the surface of possibility. Amazing innovations on the horizon will combine to enhance the driving experience of the future, making the trip safer, easier and more enjoyable. Those supporting the automotive ecosystem and making it a success have recognized the potential of Industry 4.0-on-wheels and are adapting their approaches to take advantage of it. Data is the new oil and the insight from the data is the new fuel. As they craft their solutions, OEMs are realizing the key is to not think about connectivity as a cost, but rather as a business enabler.
To learn more about connectivity management and seamlessly making it a part of digital automotive service offering, please check out how Ericsson captures the full potential of connected vehicles.


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Sewing and Pattern Making Classes
DesignerStitch School of Fashion and Design
Clothing Designers, Fashion Industry, Sewing Centre
The Designer Stitch School of Fashion and Design is dedicated to teaching design, and sewing enthusiasts, unique methods of accomplishing both basic and traditional sewing projects. Learn the principles of both design and development, and extend and sharpen your sewing skills that are needed for design, fit, and construction.
At the school we offer a series of Sewing, Pattern Making, and Design Courses and Classes, suitable for the absolute beginner in sewing, right through to those wishing to design their own garments. Our courses range from the Sewing Course- Basics and Beyond, for the absolute beginner or those wishing to advance their sewing techniques, right through to Pattern Making- Innovative Course for those who have their sewing skills in place, but who want to design their own clothes.
The aim of the school is to build your design and sewing skills in the correct progression, teaching : WHAT you should be taught WHEN you should be taught and ALL you should be taught about DESIGN, and SEWING CONSTRUCTION. Students are taught the latest tricks, shortcuts, and sewing techniques.
Day and Evening classes are held in a well equiped sewing studio, which enables the student an opportunity to design their own unique creations which express the individuality and unique creativity of the wearer.
Through the classes you will establish and extend your sewing and pattern making skills, increase your knowledge and enhance your creativity which will last you a lifetime.....and have a whole load of fun while you are doing it!!!!
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text:
This paper reports that the abundances of endogenous cardiolipin and phosphatidylethanolamine halve during elongation of the Gram-positive bacterium Listeria innocua. The lyotropic phase behaviour of model lipid systems that describe these modulations in lipid composition indicate that the average stored curvature elastic stress of the membrane is reduced on elongation of the cell, while the fluidity appears to be maintained. These findings suggest that phospholipid metabolism is linked to the cell cycle and that changes in membrane composition can facilitate passage to the succeding stage of the cell cycle. This therefore suggests a means by which bacteria can manage the physical properties of their membranes through the cell cycle.
The bacterial cell cycle is an example of binary fission that is typically both rapid and reliable. It is characterised by periods known as B, C and D1,2,3,4. The B period represents the phase between division and the initiation of replication, whereas the C period (sometimes called the replication phase) represents the phase in which the nucleolus is replicated and in which the bacterium reaches its mature size. The D period is that between the replication of the DNA and fission of the cell envelope. The B, C and D periods can therefore considered analogous to the eukaryotic G1, S and G2 phases, respectively. However, despite considerable research, little is understood about how progress from the B to D periods is controlled.
In Eukaryotes it is known that the cell cycle is controlled through the interaction of cyclin and cyclin-dependent kinases5,6,7. However, to date there is no evidence for the presence of homologues of cyclin in bacteria. Furthermore, studies of Gram-negative species have been unable to produce evidence for regulation of cell division either through changes in gene expression8 or changes in the concentration of the proteins that comprise the divisome9, 10. Existing models for the control of bacterial cell division allow for several possible mechanisms. One is based on regulation of increase in cell size (the size adder theory11,12,13). Another is that cell size is regulated in advance of division, possibly through dilution of a ‘timekeeper’ protein, though the mechanism underlying this model is not known13, 14 and it is not clear whether such proteins would control, or are rate-limiting, with respect to the cell cycle. There are also hints that cell structure is a determinant in controlling the binary fission in bacteria15. In principle, control of progress through the cell cycle through changes in cell structure could be based on both structural proteins and lipids, but knowledge of the role of lipids in this is lacking.
Recent work on lipids and their metabolism in the Gram-positive bacterium Bacillus subtilis has revealed a link between the synthesis of phospholipids and the divisome. This evidence suggests that de novo synthesised lipids are produced at the septum during fission16. Furthermore, there is evidence for an overall change in the lipid profile of the Gram-negative Escherichia coli through its cell cycle15. The latter work showed that the abundance of curvature-forming cardiolipin (CL) falls whilst that of the bilayer-forming phosphatidylglycerol (PG) increased on elongation of the cell at the C/D boundary. Evidence from biophysical studies conducted over the last half-century shows that modulation of lipid composition can have a considerable impact on the lyotropic phase behaviour of lipid systems17, 18. For example, an increase in the fraction size of phosphatidylethanolamine (PE) increases the propensity of a lipid system to assemble into curved phases rather than bilayer-like ones19, 20. This is known as increasing stored curvature elastic stress, SCES21.
Determining the relationship between lipid composition and phase behaviour complements the interpretation of lipid profiling studies. Data from studies of the phase behaviour of lipids suggest that the increase in abundance of a bilayer-forming lipid and the reduction in abundance of a non-bilayer forming lipid that occurs in E. coli on elongation, reduces SCES22,23,24,25. This is consistent with the changes in morphology of the cell during elongation because average curvature is lower in elongated cells and the evidence that the location of the Z-ring may be determined by cell membrane geometry26,27,28. This led us to the hypothesis that the production and shape of the cell envelope is involved in controlling the cell cycle of bacteria. If correct, this hypothesis implies that the topology of the plasma membrane and thus the physical properties of its lipids play a significant role in progress through the cell cycle in bacteria.
In order to investigate whether modulation of lipid composition through the cell cycle is a general feature of bacteria we measured global changes in the lipid profile of a bacterium that is evolutionarily distinct from E. coli. Listeria innocua NCTC 11288 was chosen because, unlike the commonly studied model Gram-positive B. subtilis, it does not sporulate under stress. L. innocua also has clinical relevance29, due to its biological similarity to the pathogen L. monocytogenes 30, 31.
Cultures of L. innocua in the exponential phase were stopped after elongation but before cell division (around the C/D period boundary) with the bacteriostatic antibiotic rifampicin (RIF) using an adapted15 version of an established method32,33,34. The lipid fractions (LF) were isolated from freeze-dried, pelleted cultures whose cell walls had been thoroughly digested. The LFs were profiled using a combination of solution phase 31P NMR and HR-MS/MS35. The relationship between changes in lipid composition and membrane morphology observed in L. innocua was investigated using broad line 31P NMR to characterise the phase behaviour of model systems that corresponded to the lipid compositions observed.
Culture growth and synchronisation
Cultures of L. innocua NCTC 11288 were grown in tryptic soya broth at 37 °C and the growth phase synchronised using RIF15. A growth curve was used to identify the appropriate point to administer the arrest drug (determined empirically to be 240 min (Fig. 1).
The experimental steps for preparation of samples and collection of profiling data are shown in Fig. S1. Based on their size distribution the synchrony of the untreated exponentially growing cultures was determined to be >80% (and the cell size analysis suggested that these were predominantly in the B period) (Fig. 1). The cell size was measured using cells stained with Nile Red observed using fluorescence microscopy. These data were used to establish the efficacy of RIF in arresting and holding cells at the C/D boundary and cells from such cultures were found to be significantly longer (mean = 1.92 µm, ± 0.44 µm) than those in the untreated samples (mean = 1.34 µm, ± 0.29), with only 6.7% being as short or shorter than those seen to predominate in the untreated samples (t-test, p = < 0.01). Based on these results the untreated samples were taken to be representative of cells in the B period, and the RIF-treated samples representative of cells at the C/D boundary.
We took several steps in order to avoid chemical and enzymatic degradation of lipids. After centrifugation cells were resuspended in PBS to suppress PI-PLC activity36, and a mixture of lipase inhibitors was added to suppress the activity of these enzymes ex vivo 15, 35. A mixture of mutanolysin37, lysozyme and RNAase was added to digest the cell wall and thus maximise access to the lipid fraction38.
Possible contamination of the cultures was checked for by inspecting the 1H spectrum of the FA fraction but none was found (that of Listeria ssp. 39,40,41,42 is distinct from other commonplace bacteria, such as E. coli 43,44,45,46. See Preparation of fatty acid isolates in Experimental Methods, and Fig. S2).
Isolation of the lipid fraction
The LFs were extracted using a two-step method that was designed to address the apparent short-comings of traditional methods47 (see Experimental for details). Triethylammonium ions were used to provide a counter-ion that is soluble in organic solvents and thus favours migration of phospholipid species into the organic phase. Dichloromethane was used in place of chloroform in order to minimise chemical degradation of lipid species47. Lastly, in order to maximise the extraction efficacy, the freeze-dried free-flowing powdered cellular matter was washed in a mixture of organic solvents (dichloromethane/methanol/triethylammonium chloride, 3:1:0.0005, v/v/w) before being dispersed in water and methanol, and extracted with dichloromethane. In order to test the effectiveness of this procedure, the aqueous fraction and precipitate from the isolation of the LF were freeze-dried and treated with strong acid (5% H2SO4) in order to hydrolyse any FA ester bonds and thus release any remaining FAs. The amount of FA found is treated as proportional to the mass of the lipid not collected during the isolation. The data from these measurements (1H NMR, see Experimental and Fig. S3) suggest that the procedure used in this study extracted >99% of the LF. This compares favourably with around 85% for the original Bligh & Dyer method48.
Profiling of the lipid fraction
The LFs were profiled using a combination of solution phase 31P NMR and HRMS/MS35. 31P NMR was used for quantification because it allows non-destructive and unambiguous high-resolution spectroscopy of the organically-soluble phosphorus-containing molecular species present. Lysylated lipids have not been profiled using 31P NMR before and so the shifts of lysyl-dioleoyl phosphatidylglycerol (lysyl-DOPG) were determined (Fig. S4). Representative spectra of the lipid profiling and the relative abundance of lipids in the B period and at the C/D boundary are shown in Fig. 2A. A combination of high resolution and tandem mass spectrometry were used to identify both the head groups present and the range of lipid isoforms present (Table S1 shows the lipid abundance and isoform profiles. Representative lipid fragmentation is shown in Fig. S5).
These data show that the abundance of both cardiolipin (CL) and phosphatidylethanolamine (PE) fall significantly between the B period and at the C/D period boundary. The fraction size of CL falls by around 40% whereas that of PE falls by about 60%. The absolute value for the abundance of PG varies between cultures, but does not appear to be statistically significant. This is perhaps surprising in view of the significant decreases in both CL and PE, however it is not clear why this should be. It seems unlikely that this is due to lysylation as the abundance of lysyl-PG is also not statistically significant, despite notable shifts in the absolute values. The abundance of major structural lipids CL and PE does not change substantially between cultures at the beginning and end of the exponential phase, while PG changes to some extent (Table S1). Unlike E. coli, the age of cultures of Listeria does not appear to correlate with changes in its lipid profile.
Profiling of the fatty acid residues (FARs) of the total lipid fraction did not indicate a difference in the FA profile between periods, suggesting that FA production is not linked to the cell cycle in this organism (Fig. S6). These data therefore indicate that although the lipid profile changes through the cell cycle of L. innocua it does not change with the age of rapidly-dividing cells. Furthermore, cell-cycle dependent changes are restricted to the head groups of the major structural species. Taken together, these data suggest that synthesis of the principal head groups in particular is linked to the expansion of the membrane during the B and C periods.
These data (Fig. 2B, Table S1) are consistent with the published evidence that the bulk of the Listeria LF comprises PE49 and lysyl-derivatives of PG and CL50, but is at odds with reports in which PE appears not to have been found39, 50. Conversely, phosphatidylinositol (PI) was not detected in any sample in this study despite the presence of several PLC inhibitors, contrasting with reports of the presence of PI in a closely-related strain of Listeria 42, 51. We note that the lipid profile of biological samples can be influenced by myriad factors, including the medium used, how cultures are incubated and handled, how the LF is isolated, handled and profiled, and even the activity of endogenous lipases ex vivo 47.
Lyotropic phase behaviour of model membrane systems
The evidence for modulations in the abundance of CL and PE during cell elongation raises the question of whether membrane properties change with these changes in composition. Studies of the phase behaviour of PG, PE and CL over the last half-century have shown that they have distinct properties. PE’s spontaneous curvature is negative19, 52,53,54, as is CL’s but only in the presence of a high concentration of divalent cations23, 55, 56. PG’s phase behaviour is dominated by lamellar phases, suggesting that its spontaneous curvature is negligible24, 25, 57,58,59. Modulation of the abundance of PE and CL (with a low concentration of divalent cations) is therefore a change in the abundance of two major structural lipids with contrasting spontaneous curvature. That these changes in abundance occur through the cell cycle therefore suggests that the stored curvature elastic stress of the membrane also changes through the cell cycle. We investigated this by modelling the behaviour of two-lipid mixtures using broad line 31P NMR.
Model systems comprised the di-oleoyl isoforms of PG and PE (DOPG and DOPE), and tetra-oleoyl isoform of CL (TOCL). Temperature scans of the individual lipids revealed the established behaviour, i.e. systems comprising either TOCL or DOPG showed fluid lamellar (Lα) phase over the temperature range studied (Fig. S7A,B), with DOPE undergoing a transition from Lα to inverse hexagonal (HII) only above 273 K (Fig. S7C).
Mixtures of DOPG with TOCL show a strong preference for Lα, even at relatively low temperatures (273 K, Fig. 3A; Fig. S8A–C), suggesting that both lipids confer fluidity on systems in which they are present. The Lα phase also dominates across most of the concentration range in mixtures of DOPG and DOPE at 273 K (Fig. 3B), but higher temperatures favour curved phases (e.g. 310 K, Fig. 3C; Fig. S9). These data indicate that PE confers stored curvature elastic stress on PG systems. In particular, a reduction in PE from 36% to 18%, similar to that on elongation of L. innocua, reduces the ability of that system to form curved phases, suggesting a significant reduction in SCES. The Lα dominates in mixtures of DOPE with TOCL where the concentration of DOPE is below 50% (273 K, Fig. 3D). This also indicates the preference for lamellar phase(s) by TOCL at low concentrations of divalent cations (Fig. S10A), and curved phases by PE (Fig. S10C). Thus, although the fraction of CL shrinks on elongation of L. innocua, the increase in that of the DOPG ensures that the reduction in the abundance of PE reduces SCES.
However, it appears that doping DOPE with rather small quantities of DOPG (22-13:1, 4.3-7.1%) or TOCL (~36-12:1, 2.7-7.7%) allows the formation of curved phases at 273 K, particularly the HII phase (Figs 3B and S9, and Figs 3D and S10). Importantly, the phase transition from fluid lamellar to the HII phase occurs in DOPE around 10 K higher than this, at ∼ 281K60. This suggests that the presence of small quantities of these lipids can relieve the packing stress of the HII phase.
Data from a mixture of lysyl-DOPG with TOCL suggest that lysyl-DOPG has similar fluid properties to DOPG (Fig. S8C and D) but does not reduce the packing stress of DOPE systems as DOPG does (Fig. S9E and F). Notably, the behaviour of DOPE:lysyl-DOPG (22:1, Fig. S9F) is very similar to that of pure DOPE (Fig. S7C), and both are distinct from DOPE:DOPG (22:1, Fig. S9E). In the latter, the less abundant lipid appears to reduce the packing stress of the inverse hexagonal phase. This suggests that the role of lysyl-DOPG in the membrane differs from that of PG and may be related more to modulations in SCES and/or lipid packing than to fluidity.
The evidence from this study indicates that the modulations in lipid composition in L. innocua that occur between the B period and C/D boundary are statistically significant. The behaviour of model systems suggests that the modulations in lipid composition on elongation of L. innocua represent a reduction in the SCES of the membrane (observed as a reduction in size of PE fraction, Fig. 3B) but without a reduction in its fluidity (with an increase in PG and despite a decrease in CL; Fig. 3A and D). This suggests that the physical properties of the membrane are linked to the cell cycle.
The behaviour of model lipid systems observed in this study supports the conclusion that changes in lipid composition favour changes in morphology of the membrane on a cellular scale. Comparison of the lipid profile and distribution between different rod-shaped bacteria (such as E. coli and B. subtilis) and between rod-shaped and spherical bacteria (such as Staphylococcus aureus) are therefore of interest in understanding the cellular structure of prokaryotes. However, comprehensive cell-cycle-based lipid profiling of prokaryotes is lacking at present. What is known, is that E. coli and B. subtilis have very different distributions of lipids in their membranes, which are linked to differences in proteins associated with cell division, and therefore cell shape is not expected to be the only factor that determines this change61.
We note that the modulations in lipid composition of the Gram-positive L. innocua are analogous to those reported for the Gram-negative bacterium, E. coli. In the latter organism, the abundance of the bilayer-forming PG increases during its elongation (from ~1:22 to ~1:16 against PE), with a reduction in the abundance of CL (from ~1:16 to ~1:36 against PE)15. Physical data from the present study suggest that there is also a reduction in the tendency to form the hexagonal phase (lower SCES) in E. coli, without a reduction in fluidity on progression from the B period to the C/D boundary. This indicates that E. coli has adopted the same pattern with respect to modulation of lyotropic phase behaviour between the B period and C/D boundary as seen in L. innocua.
This remarkable similarity between two bacteria that are quite separate in evolutionary terms raises the question of whether such modulations are important for the structure of rod-like cells in general. However, it is not clear from these data whether this trend is limited to cells with a cell wall; further work is required to establish what effects modulations in lipid composition have in other cell types.
It is appealing to speculate what further effects the modulations in lipid composition may have on membrane properties. The folding and function of proteins of both bacterial and eukaryotic origin have been shown to be sensitive to lipid membrane composition62,63,64, and to SCES and packing defects65, 66. This coupling between the membrane’s physical state and the proteins’ folding can also manifest itself in problematic rather than functional ways. For instance, there is evidence that charged head groups such as those of PG and phosphatidylserine enhance the membrane binding and mis-folding behaviour of α-Synuclein67, and that packing defects are involved in promoting mis-folding of a protein linked to Parkinson’s disease68, 69. Particular constellations of ganglioside and cholesterol have also been proposed to promote both membrane binding and mis-folding in α-Synuclein and β-amyloid peptide70.
It is clear that further work is required to fully characterise the relationship between lipid composition and distribution in bacteria71 and in eukaryotic72 cells. Data such as that acquired in this study indicate the extents to which the lipid composition and phase behaviour vary with the cell cycle. These alterations may in turn have an impact on the structural and functional properties of the membrane they comprise.
Reagents and chemicals
Growth medium, solvents and fine chemicals were purchased from SigmaAldrich (Gillingham, Dorset, UK) and used without purification. PhosSTOP tablets were purchased from Roche (Welwyn, Hertfordshire, UK) and stored at 4 °C. Purified lipids were purchased from Avanti Polar lipids Inc. (Alabaster, Alamaba, US) and used without further purification.
Cultivation of L. innocua
The growth kinetics of this bacterium in tryptic soya broth (TSB) was characterised over 24 h (Fig. 1). The preparation of one set (n = 1) of cultures is described. Each sample comprised three cultures in order to ensure that enough material was produced and to account for differences between populations. Thus for each biological replicate, 3 × 3 × 1 L cultures were grown in unmodified TSB that had been autoclaved. Mini-cultures (10 × 10 mL) were incubated (16 h, 37 °C, orbital incubator, 250 r.p.m.,) before inoculating 9 × 1 L to give cultures of OD620 ~0.03. After incubation (37 °C, 240 min, orbital, 250 r.p.m., average OD620 0.4), three cultures were mixed together to make the 4 h control time point and cells harvested (fixed-angle rotor, 6k × g, 10 min, 4 °C). Three cultures were treated with RIF (50 mg/L final concentration, added as methanolic solution of 50 mg/mL). After 360 min h, the three remaining control cultures were mixed and harvested together (making the 6 h control time point which is equivalent to the point at which the cultures treated with RIF are arrested), and after 480 min the remaining three cultures treated with RIF were harvested together.
Fluorescence and light microscopy
Cells were photographed after harvesting and before the administration of lipase inhibitors. Cells were stained with Nile Red as previously described73,74,75. Briefly, cells were collected by centrifugation (fixed-angle rotor, 6 k × g, 10 min, 4 °C), washed once with PBS and resuspended in PBS at OD620 ~2.0. Nile Red (2 µg/mL, sample volume 1 mL, dye stock solution in DMSO) was added and the sample (1 mL) allowed to stand at room temperature (20 min). Cells (60–80 per picture, 180–200 per sample) were measured from at least three images for each culture/time point each from four independent cultivations (i.e. (~70 × 3) × 3 × 4). The mean and standard deviation were calculated for each and an independent student t-test was used to determine statistical significance (p = < 0.01). The null hypothesis was that the lengths of the bacteria are the same for each sample. Images were taken using a Leica DMI6000 B microscope equipped with a Leica DFC350 FX camera and processed with (the 3D deconvolution function in) Leica Application Suite, Advanced fluorescence 1.7.0 software with AF6000 configuration and open source ImageJ software.
Isolation of lipid fraction
Fresh cell pellets were resuspended immediately (PBS, 5 mL) and treated with PhosSTOP (1 tab/10 mL final concentration, 10 × stock dissolved in PBS), 2-butoxyphenylboronic acid (BPBA, 2 mg/mL final concentration, ethanolic stock solution 100 mg/mL) and a stored mixture of mutanolysin (50 µg), lysozyme (10 mg), RNAase (200 µg) (volume of aliquots 1 mL, glycerol/PBS 1:1, stored at 193 K). The total volume of the resulting solutions was 10 mL per sample and they were agitated gently in air tight falcon tubes (50 mL, gel staining table, 16–20 h) before being frozen (193 K) and freeze-dried. The freeze-dried material was stored at 253 K until isolation of the lipid fraction.
The free-flowing powdered cellular matter was mixed with dichloromethane/methanol (20 mL, 3:1, 0.5 mg/mL triethylammonium chloride, TEAC) and sonicated (sonication bath, 5 min) before being centrifuged (5 min, 5.25 k × g). The organic solution was decanted and retained. The pellet was mixed with more of the same dichloromethane/methanol mixture (20 mL, 3:1, 0.5 mg/mL TEAC), agitated, centrifuged and the solvent decanted. The pellet was then resuspended in a mixture of dichloromethane and water (20 mL, 1:1, separating funnel) and diluted with sufficient methanol to make a stable uniphasic solution (~40 mL), and agitated briefly. The mixture was then made biphasic by addition of dichloromethane (20 mL). The dichloromethane solution was removed and the aqueous solution washed (dichloromethane, 20 mL). TEAC was added to the remaining aqueous solution (final concentration of 2 mM, 2 M stock) and the aqueous solution washed with dichloromethane (2 × 20 mL). The combined organic solutions (~130 mL) were filtered through filter paper and concentrated in vacuo before storage under nitrogen at 253 K.
Isolation of residual lipidic material
The aqueous phase and precipitate produced during the lipid isolation (above) were freeze-dried together and then treated with acid (H2SO4, 5%, 20 mL, gentle agitation, 24 h, r.t.p.). The reaction mixture was quenched (NaHCO3) and freeze-dried. The free-flowing powder was washed twice with chloroform. The organic phases were combined and concentrated in vacuo. The remaining material (~8 mg/120 mg LF isolated) was dissolved in deuterated chloroform (650 µL) before commencing 1H NMR. The integrals of each spectrum were calibrated according to the α-CH2 signal (2.3–2.4 ppm) present (Fig. S4).
Solution phase 31P NMR spectroscopy
Lipid films (10-15 mg) from cultures of L. Innocua were dissolved in the CUBO solvent system15, 35, 76 (500 µL/sample). Data acquisition was similar to published work15, 35 but using a Bruker 400 MHz Avance III HD spectrometer equipped with a 5 mm BBO S1 (smart) probe operating at 298 K for all data except that shown in Fig. S5. 31P spectra were acquired at 161.98 MHz using inverse gated proton decoupling, with 2048 scans per sample and a spectral width of 19.99 ppm. An overall recovery delay of 6.5 s was used which gave full relaxation. Data were processed using line broadening of 1.00 Hz prior to zero filling to 19428 points, Fourier transform and automatic baseline correction. The data for Fig. S5 were acquired on a Bruker Avance III 800 MHz spectrometer, equipped with a QCI cryoprobe probe. Acquisition used inverse gated proton decoupling. Spectra were averaged over 1312 transients with 3882 complex points with a spectral width of 14.98 ppm. An overall recovery delay of 8.4 s was used. Data were processed using an exponential line broadening window function of 1.5 Hz prior to zero filling to 32768 points, Fourier transform and automatic baseline correction. All spectra were processed and analysed using (the dcon function in) TopSpin 3.2.
Preparation of fatty acid isolates
Representative samples used for head group profiling were dried in vacuo (high-vac pump) before being diluted (H2SO4 97% in methanol, 1:20 v/v final concentration, 5 mL) and heated under intermittent agitation (4–6 h, 50 °C). The reaction mixture was quenched (NaHCO3, 1 g) before being diluted (H2O, 30 mL; diethyl ether, 20 mL). The mixture was shaken vigorously in a separating funnel before the organic fraction was collected. The aqueous solution was washed (diethyl ether, 20 mL). The organic solutions were combined and dried in vacuo. The resulting oil (5–10 mg) was dissolved (CDCl3, 500–600 µL) and subjected to 1H NMR spectroscopy. δH (500 MHz, 16 scans, CDCl3, calibrated to 7.24 ppm) 3.6 (singlet, RCH2CH2COOCH 3 ), 2.3 (triplet, J = 7.6 Hz, RCH2CH 2COOCH3), 1.6-1.5 (m, RCH 2 CH2COOCH3), 1.4-1.0 (m, methylenes), 0.9-0.8 (m, methyl groups) ppm.
Samples were prepared and measurements taken according to published methods15, 35. Accurate mass LC-MS and MS/MS was performed on an OrbiTrap Dionex 3000, (Waltham, MA, USA). Dry lipid mixtures were dissolved in dichloromethane/isopropanol 1:1 and an inject volume of 10 µL was used. Analytes were separated chromatographically on a UPLC C18 column (1.7 µm particle size, Waters) at 40 °C at a rate of 0.4 mL/min. Mobile phase A consisted of 0.1% formic acid in water at pH 6.0, and mobile phase B was 56% acetonitrile, 40% isopropanol and 5% water with 0.1% formic acid. Ions were monitored in positive mode (m/z range 300-2000, resolution 140,000). Selected peaks were fragmented (normalised collision energy set to 36) and MS2 collected with a resolution of 17,500.
Analyses consisted of searches for lipid isoforms in MS1 spectra, followed by fragmentation (MS2) for appropriate m/z values. Lipid isoform searches were restricted by head group (PG, PE, PA, PS and CL), FAR (chain length from 10 to 20 C, up to three double bonds) and adduct (H+, Na+, K+, NH4 +). The search was augmented to include lysyl and lyso- isoforms. Matches were compared (see Fig. S6 for example). The software-generated scores were checked manually. Analyses were performed using Thermo Xcalibur 3.0.63 and recently-developed software by Kochen et al.77. Original code for non-standard head groups was written by us in Matlab R2015b.
Broad line 31P NMR sample preparation
Lipid mixtures were made in solution (dichloromethane, 10–15 mg/mL) and dried to a lipid film in vacuo. Dried lipid films were dispersed in aqueous buffer (NaCl 100 mM, tris 50 mM, CaCl2 2.5 mM, MgCl2 2.5 mM, pH 7.4; 30:1 v/w) with sonication and agitation. Deuterium oxide (10% v/v against buffer) was added and the mixture agitated and then freeze-thawed 8–10 times. Samples were then stored at 193 K, transported at ~295 K and were stored at 253 K before running.
Broad line 31P NMR spectroscopy
A Bruker Avance III HD 400 MHz DRX spectrometer equipped with a Bruker BBO S1 (smart) probe was used for proton-decoupled broad line 31P NMR experiments. Experiments were performed at 161.98 MHz with an inverse-gated pulse sequence with proton decoupling during acquisition, spectral width of 200.44 ppm, acquisition time of 0.299 s, pre-scan delay (DE) time of 20 µs, receiver gain of 203 with 19428 data points and 2048 scans per spectrum. The duration of each scan, acquisition time and relaxation delay, was 2.332 s. Spectra were processed using TopSpin 3.2 and 3.5 with line broadening of 50 Hz, phase correction and automatic baseline correction (abs).
Broad line 31P NMR temperature scans
Samples were brought to and held at the desired temperature for 15 min before acquisition. Temperature scans consisted of acquisitions at the following temperatures: 298, 273, 293, 298, 310, 318, 273, 310 K. The latter two temperature points were used to assess the reliability of the equilibration time used (15 min). Acquisitions at 338 K were carried out separately with equilibration and acquisition at 310 K immediately beforehand. Stackplots of spectra were prepared manually from individual traces processed as above.
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The Research Council of Norway is acknowledged for funding (grant application ES528542), and for financial support through the Norwegian NMR Platform, NNP (grant 226244 / F50). The authors wish to thank Drs Fedor Kryuchkov and Marc Niere for assistance with MS and microscopy, respectively, and SF would also like to thank Drs J. Loewe, S. Liddell and T. Allers for helpful discussions. The authors thank Drs Ø. Larsen and A.E. Lewis for critical review of the manuscript and acknowledge Dirk Petersen (NNP UiO node) and Olav-Audun Bjørkelund (NNP UiB node) for their help in facilitating the work.
The authors declare that they have no competing interests.
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Furse, S., Jakubec, M., Rise, F. et al. Evidence that Listeria innocua modulates its membrane’s stored curvature elastic stress, but not fluidity, through the cell cycle. Sci Rep 7, 8012 (2017). https://doi.org/10.1038/s41598-017-06855-z
Analytical and Bioanalytical Chemistry (2021)
Analytical and Bioanalytical Chemistry (2020)

domain: literature


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What you need to know
- Call of Duty: Warzone in the battle royale mode for Call of Duty: Modern Warfare.
- It currently supports up to 150 players in a battle royale match.
- A new update is introducing a limited time 200 player match.
- This update is scheduled to go live at 11:00 p.m. PT tonight.
If you thought Call of Duty: Warzone was done growing, think again. Today, Activision announced a new limited time 200 player mode that will be included in an update.
This update is scheduled to go live at 11:00 p.m. PT tonight. Verdansk was previously limited to 150 players in a match, so the addition of 50 extra players should shake things up, even for battle royale veterans.
Call of Duty: Modern Warfare has been a phenomenal success, continuing to sell well and top sales charts months after release. Combined with the free-to-play standalone release of Call of Duty: Warzone, this battle royale just continues to grow with no sign of slowing down.
Call of Duty is an annual franchise, never missing a new entry in a new year. With that said, we've almost hit July 2020, which will be the latest Activision has ever waited to showcase a new Call of Duty title. Prior rumors indicated 2020's game would be called Call of Duty: Black Ops Cold War, which was suffering from reported development problems.
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AC Podcast 492: TikTok/WeChat Ban; LG Wing; Pixel 5s; PS5
Daniel and Jerry are joined by Alex Dobie and Joe Maring to make sense of the impending (at the time of recording) TikTok and WeChat bans in the U.S. The crew also preview the LG Wing and discuss Apple's recent Watch and iPad announcements in comparison to Android-compatible watches and the Galaxy Tab S7. Alex also reports on the rumored Pixel 5s. Plus, the next-gen consoles are coming...
Foldables are finally good enough to actually spend money on
Foldable phones have come a long way in 18 months, and now with the Galaxy Z Fold 2, we have no major flaws, no shortchanged specs and no hurdles still to overcome. Now is the tipping point when foldables start to actually become worthy of your wallet.
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The Oculus Quest 2 was announced at Facebook Connect 2020. Here's how to preorder the hottest new wireless VR system around!
Make the perfect play with these PS4 sports games
Here are the best sports game that can give you some of the thrills of competition when you can't play outside. They can also let you imagine what the sports of the future might look like or even give you a workout at home. Play competitively or casually with friends.


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Social network analysis of the movement of poultry to and from live bird markets in Bali and Lombok, Indonesia.
J Kurscheid, M Stevenson, PA Durr, J-ALML Toribio, S Kurscheid, IGAA Ambarawati, M Abdurrahman, S Fenwick
Transbound Emerg Dis | Published : 2017
Highly pathogenic avian influenza H5N1 has resulted in large losses to the Indonesian poultry sector. Evidence suggests that live bird markets (LBMs) play an important role in the epidemiology of the disease. Knowledge of the frequency and type of contact between the various poultry market players should allow animal health authorities to develop a better understanding of factors influencing virus transmission between Indonesian villages. A questionnaire-based cross-sectional survey was conducted in 17 LBMs on the neighbouring Indonesian islands of Bali and Lombok to investigate the movement patterns of poultry to and from markets. Using social network analyses, a network of contacts was cre..View full abstract
Awarded by Australian Centre for International Agricultural Research

domain: biology


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Just days after an order by a local court, the Saidabad police in Hyderabad registered a case against All India Majlis-e-Ittehadul Muslimeen (AIMIM) MLA Akbaruddin Owaisi.
Owaisi has now been booked under Sections 153(A) (Promoting enmity between different groups on grounds of religion), 153(B) (Imputations, assertions prejudicial to national-integration) and 506 (Criminal Intimidation) of the IPC.
The case has been registered over alleged inflammatory remarks made by Owaisi at a public meeting on July 23 in Karimnagar town. Akbaruddin Owaisi said, "The Rashtriya Swayamsevak Sangh (RSS) can't do anything. They can't even pluck a strand of hair," and went on to refer to his infamous '15 minute' comment.
On December 22, 2012, Owaisi had addressed a massive rally in (then Andhra Pradesh's) Nirmal town, where he had made a comment about "removing the police for 15 minutes off the streets", suggesting violence would erupt between Hindus and Muslims. He was arrested in January 2013 on charges of hate speech, and granted conditional bail after 40 days in prison.
Earlier this month, the 14th Additional Chief Metropolitan Magistrate court had issued orders to the Saidabad police, to investigate a complaint by an advocate associated with the Bharatiya Janata Party (BJP) and register a case and file a status report by December 23.
The advocate had moved court, stating that the police were not investigating his complaint. In his petition, he also sought that Owaisi's conditional bail be cancelled.
Owaisi's remarks earlier this year had also resulted in a similar complaint being filed in Karimnagar town at the time. The Karimnagar 3-Town police has also booked him under the same sections as the Saidabad police and is investigating the issue.
As a video of the recent speech also went viral, Akbaruddin Owaisi issued a statement on July 26, where he said, "My speech doesn't constitute any offence; I have not contravened any provision of law.”
Akbaruddin Owaisi is the younger brother of Hyderabad MP and AIMIM President Asaduddin Owaisi.


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Eliot Spitzer was the last to speak here tonight, and it was his speech that made the greatest reach towards grandeur.
“Today was not a victory of one candidate or one Party but of all those irrepressible optimists who have dreamed of a resurgent New York,” said Spitzer, who vowed to work to make New York “once again the greatest state in the nation.”
He stepped back from the podium to kiss his wife. He marveled at his parents’ 61-year marriage. (“As they say on my favorite TV show, ‘Booyah.'”) And he quoted Alexander Hamilton and Walt Whitman.
He built up to a crescendo, noting that his oft-stated conviction that everything changes on Day one was “now the promise of a governor” and added, “I will never break faith with those who have invested their hopes and dream on this great state.”
With that, the entire Democratic ticket (minus Alan Hevesi) climbed on stage, raised their arms and clapped to Start Me Up. And the rank-and-filers headed for the exits.

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Most of the pages I made for our Round Robin book, are "low color". But every now and then, I slipped in some floral tones, just because I thought it needed a little something. This page for Beth, is a tissue pattern paper background, glued down nice and wrinkly, then rubbed with an ink pad.
A ruffle of pattern paper was glued across the top, and a swath of lace was added near the bottom of the page. The B for Beth is drawn in glitter, onto a very old, hand written letter in French. I centered it inside a post card, with a circle cut out, layered it onto text from an old sewing magazine from the 20s that I'd covered a tag with.
The ribbons hanging down have glass buttons tied to a couple of their ends.
The theme of the Round Robin was "Her Life in Stitches", and I simply don't sew. All of my pages are glued, while the other women spent time doing lovely stitch work. I tried to use the theme by incorporating some sort of sewing item on each page. I tended to rely on buttons, lace, and ribbons quite a bit, I'm afraid.
Beth is one of the few people I've known who uses both sides of her brain to the fullest. How many ultra artistic accountants do you know? Her head is quite ambidextrous.
So, I thought I'd show both sides of her in her pages, creative and pretty stuff along with the business side.
This one has an envelope and report cards from a business college and is dated 1918. The background is a great old, gold leaf wallpaper.
Since the books all need to be sent off to Somerset soon, (how exciting!!) I hurried and made a few pages up for each person in the swap. All the pages were done in the same day, so a lot of the ideas and supplies were duplicated, but none were exactly the same. Everyone got an initial, Sandra's is done in pearls.
My own initials went on the inside of the back cover of my book. Since I'm not a sewer, my book is dedicated to my Mom, who was. Her photo is on both the front and back covers. Under her picture, I layered a page from a journal that I'd found that was signed, "Martha something Downey".
Mom's name was Martha Downey! Not that common of a name, especially with the "e" in Downey. I used the bottom of her photo to cover up the incorrect middle name.
Another unifying element in the pages I made the other day, is this jeweled and beaded fabric. I bought a flapper style dress that was encrusted with beads and rhinestone trim for only 12 bucks! I think everyone who was shopping at the antique mall heard my squeal when I came across that gem. It was tattered and falling apart, so that took all the guilt out of chopping it to pieces.
For this pocket, I cut around the beading pattern, then glued it to a matching piece of card stock.
Its attached to a background of white, flocked wallpaper. When I make a pocket, I always run a bead of glue along the bottom and sides, right at the edges, then slightly bunch it up, before pressing it into place. The bunching gives it a bit of a pouch effect, so that you can put items into it easier. Its not all that noticeable, just enough to be useful.
To fit the stitches theme, I slipped a dress form tag into the pocket. The tag is one that I made for my dress form tutorial, the class is about making a miniature dress form to decorate, but I also included patterns for collage too.
The tag I added to this pocket is a regular shipping tag, spritzed with two shades of Glimmer Mist, then inked around the edges. I glued a high fashion lady cut from a very old magazine onto it, added some ribbons.
Its been such fun to be a part of this swap and have a different book arrive each month in the mail. I'll miss being a part of it.
Two boxes full of goodies for my "Peeps" attending our workshop/party in April came from her this week.
Laurie took the time to actually create little gifts for all of the women, plus gave them some vintage goodies to work with, and packaged them up really cute.
But that wasn't the end of it, she also asked the author of this book, that she collaborated on, to send a copy for each lady! I can't wait to peek into one. I can always use some selling tips. I wonder if one of them, is to get off your behind and get busy and do it, not just talk about filling the shop up?
If you'd like to be a sponsor, please let me know, the ladies will be thrilled to open their gift bags.
Or if you'd like to attend, we have three spaces open. There were only two, but due to an illness, one chick had to leave the nest.