text:
You may want to discuss or assignthe following article:
Rucker, D. R., & Petty, R. E. (2003). Effects ofaccusations on the accuser: The moderating
role of accuser culpability. Personality and Social PsychologyBulletin, 29, 1259-1271.
The authors report on an interesting topic (the benefits ofaccusing others), describe two interesting 2 X 2 between-subjects factorialexperiments, do a great job of showing how study 1 follows from the research ofRucker and Pratkanis (2001) and how study 2 follows from study 1, and proposesix different ways to follow up on their research (three of which would be easyfor students to do). In addition, the article is easy for your students toobtain (students who buy the book can get it by using the Infotrac® subscriptionthat comes with Research Design Explained),and the article is relatively easy to students to read (to make it even easier,see Table 1).
The article starts by pointing out that research has shownthat attacking others’ characters (e.g., negative ads) works. The article then points out that researchhas not extensively studied the effect of making accusations on theaccuser’s reputation. Then, the authors report on an article
The effects of the pot calling thekettle black. Personality and Social Psychology
Bulletin, 27, 1494-1507.
showing that an accused person could benefit by
accusingothers. This effect was called “projection.” The authors also tryto
explain why the projection paradigm obtains the opposite results (theaccuser
being perceived as less guilty) as what would be obtained with“spontaneous
trait transference” paradigm (the accuser being seenas more guilty because the
accuser is associated with those accusations ). Theauthors focus on two
differences between the paradigms: (a) accusers in“projection” paradigm may
actually be guilty and (b)
participants in the “projection”paradigm process the information more deeply
than participants in the“spontaneous trait transference” paradigm. The authors
then suggesta person who is suspected of being dishonest may benefit by
accusing others ofbeing dishonest because that accusation shows that the person
actually valueshonesty, but that a person who is assumed to be honest
doesn’tcommunicate anything new about that person’s honesty by accusing
someoneelse of being dishonest.
Inexperiment 1, “a 2 (employee culpability: culpable or not culpable ) X
2(accusation: present or absent) between-subjects factorial” experiment,the
authors tested the hypothesis that culpability would moderate theaccusation
InExperiment 2, the authors replicated Experiment 1 and tested “whetherperceived work ethic mediates the effect of accusations on performance ratingsfor culpable individuals but perceived friendliness mediates the effect ofaccusations on performance ratings for nonculpable individuals.” Procedurally, the main difference fromExperiment 1 was adding two 7-point questions about the employee’s workethic and two 7-point questions about the employee’s friendliness.
When analyzing the results from thesame scale used in Experiment 1, the crossover interaction obtained inExperiment 1 was replicated. When analyzing the results from the new“work ethic” scale, the authors found that making accusationsboosted the culpable employee’s perceived work ethic, but makingaccusations did not boost the nonculpable employee’s perceived workethic. When analyzing the results from the new “friendliness”scale, the authors found that making accusations made the nonculpable employeeappear less friendly, but that making accusations did not make the culpableemployee appear any less friendly. Regression analyses supported the idea thatperceived work ethic mediated performance ratings for culpable employeeswhereas perceived friendliness mediated performance ratings for nonculpableemployees. The article concludes by stating some practical implications of theresults (criminals and dirty politicians may benefit by making accusations) andby outlining six lines of potential followup research.
Helping Students Understand the Article
Tips, Comments, and Problem Areas
“boundary conditions” of a tactic means conditions under which the tactic will not work.
“exculpatory”: proving that someone is not guilty of the misdeed
as discussed in Chapter 2, “moderators of an effect” would be variables that reverse, neutralize, weaken, or strengthen the effect. Knowing the moderators allow us to know under what circumstances the tactic will be effective. “Mediators” refer to what mechanisms (in this case, beliefs) are responsible for the effect. Knowing the mediators allows us to know how and why the tactic works.
“Nonculpable”: Not guilty, not responsible for the bad deed.
“Culpable”: Guilty, responsible for the bad deed.
“mundane realism” refers to the degree to which the study, or tasks that the participant performs during the study, resemble a real life situation.
Cornbach’s a of .84 indicates that the items on the scale were internally consistent (see pp. 101-104 in the text).
M is an abbreviation for mean, SD for standard deviation ( a
measure of spread)
If you have trouble understanding the graph (Figure 1), the last two sentences of the results section explain it to you.
“asymmetry”: difference—in this case, it refers to making accusations having one effect when a person is innocent but another effect when a person is guilty.
Self-handicapping: giving oneself an excuse for performing poorly (e.g., not studying for a test)
You will not understand every word or term of the “Mediational analyses” subsection of the Results section. You will probably not know what regression is, what the Sobel mediation test is, or what the statistics in parentheses mean. However, you can understand the essence of the Mediational analyses subsection because you don’t need to know those terms to understand what the researchers found.
“parsimoniously’: simply, economically, and elegantly.


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“Process saves us from the poverty of our intentions.”
I am currently reading Seth Godin’s new book ‘The Practice’. More about that when I actually finish it. I was struck by the quote in the front of the book from Elizabeth King shown above and went on a hunt to find more about her. Frustratingly I couldn’t find the exact source of the quote but many people who attribute it to her but the journey was fun.
Elizabeth King is a sculptor who taught at Virginia Commonwealth University for 30 years before retiring to focus on her work. She is best known for her figurative sculpture that she combines with stop motion animation. Perhaps remarkably she was creating the articulated sculptures that were perfect for stop motion animation long before she had the idea for animating them.
I am always fascinated to watch artists at work, regardless of the medium that they work in. The behind the scenes look that you get in Olympia Stone’s documentary “Double Take’ is a real treat in this regard. It shows Elizabeth at work in her Church Hill studio as well as installing her work at various shows.
What is evident in the documentary is her attention to detail. She really cares about getting the sculpture right, capturing the imperfections in joints on the hands of her sculptures so that the fingers end up a little wonky. I thought she was describing my fingers as she described this detail in the documentary. She’s clearly looking and not just going through the motions and means that her sculpture stands up to the closest of scrutiny.
Of course her attention to detail doesn’t stop with the creation of the work. It’s interesting to see how much attention she puts into the installation to make sure that the pose of the sculpture is correct and that the lighting is just so to provide the correct emphasis to the face and hands. This is what separates masters from the journeymen.
I often fall into the trap, as I suppose many people do, of being generally dissatisfied with the work that I’m producing. I make images that I like just often enough to keep me engaged but it can be tough to keep going especially when we’re surrounded by an onslaught of great work on social media.
The guitar teacher Tomo Fujita tells his students ‘Be Kind to Yourself, Don’t Compare, Don’t Expect Too Fast, and Don’t Worry.’ Good advice for anyone whether they are trying to learn a new skill or to be creative.
The other advice that I turn to when I’m struggling is what Ira Glass said about ‘The Gap’ (see video 3 below). He’s describing the difference between what you know is good and want to be able to do and what you’re currently able to achieve.
Check out the illustrated video below.
The solution of course is to do a lot of work. Bang it out even if you don’t feel like it. Just keep going. You will get better, you will evolve and you will close the gap.
Checkout the full interview ‘Ira Glass on Storytelling’ in the following videos. This should be required viewing for anyone in the creative arts.
When I was thinking about what I had learned in the Portfolio Development class with William Neill, one of the things that I was reflecting on is how much you can learn from the other students. A good group that are freely sharing their previous experiences, understanding and viewpoint can really support your growth.
One of the students in the class turned me on to Lenswork – a magazine that I had never heard of and certainly wasn’t carried in the local bookstores at the time. I eventually ended up getting a subscription so that I could see what it was all about. Lenswork is a bimonthly magazine that emphasizes photographs not gear and is exquisitely printed – book quality printing. From what I understand the emphasis on photographs is very similar to the principals that were at the heart of Aperture Magazine when it was established and under the guidance of Minor White. To see some of the early issues check out this anthology.
The editor, Brooks Jensen, is an accomplished photographer and his work can be found at his personal site, Brooks Jensen Arts. The image above is from Brooks’ first Winter Trees portfolio – you can download a pdf of the portfolio here. I continue to be fascinated by this image – it has a depth to it, a three dimensionality, that I have not experience in any other photograph. I was fortunate enough to be able to get a print of this image when Brooks was still selling individual images and it is even more stunning in the flesh, as it were.
Brooks’ thinking about photography that he shares through his writing and podcast have had a profound impact on my thinking about the ‘photographic art life’. He makes really great points about in his article about what size should editions be, has suggested multiple ways of presenting your work to your audience including, Folios and Chapbooks and additionally was an earlier adopter of PDFs. I have learned a tremendous amount from Brooks and think you would too. Go take at look at Lenswork, Lenswork online and Brooks Jensen Arts. Listen to the interview with Brooks in the interview below.
I learn all kinds of stuff by listening to my kids talk to their friends. Some of it immediately, some of it takes me a while to unravel.
It’s snowboard season here in New England which means trips to and from the slopes with groups of high school age kids for me. They are very much into making videos of their exploits to get feedback on how they are doing with learning tricks but also to post on social media. ‘Doin’ it for the ‘gram’ was the phrase used in a rather derogatory tone.
That got me thinking about what I post on social media and why and what are my expectations. I am playing a game with my posts on Instagram, or at least I have some simple rules that I am following. Must be shot and edited on my phone. No wireless transfer of files from any of my ‘fancy’ cameras, no transfer of files from the phone onto my computer for editing. All has to be done on the phone. Also the apps that I use can’t be mobile versions of Lightroom or Photoshop.
This of course is less of an amazing feat now that the phone cameras are so incredibly capable and produce high-res files. It goes without saying that phones themselves are way more powerful computers than the first desktop computers I worked with but then I grew up with a single rotary dial phone in the house if you catch my drift.
The result is a set of photos that are in effect sketches or studies. I’m trying out composition ideas using the phone and quickly processing using filters and effects in a small number of apps – typically Snapseed and VSCO. I’m often accused of being heavy handed with the processing – probably true, certainly lacking the kind of finesse possible on a desktop. Are these the high impact, portfolio best images that will garner lots of ‘likes’ not at all. But that’s not the point.
I’m having fun playing and don’t mind letting people look over my shoulder see my sketchbook develop.
In the first 30 years of your life, you make your habits. For the last 30 years of your life, your habits make you. — Hindu saying that Steve Jobs was fond of
I tend to circle topics until I get a satisfactory answer – something that makes sense to me, is actionable or is a definitive end. It’s a funny trait that I didn’t realize I did until someone pointed it out to me recently. One of those topics is how to learn. I’ll certainly come back to this a few times here.
I was thinking about a story that I read in the book Art & Fear about a pottery class that was split into two. One group was told that their grade would be based on the quantity of work that they produce during the semester while the other group would be graded on the quality of work they produced. At the end of the semester the group that produced the most work also produced work of a higher quality. The act of making, making mistakes, correcting and making again had lead to a deeper understanding.
How can we apply this thinking to our photography to push ourselves forward? I am contemplating a project where I would post an image a day to Instagram and then review my progress at the end of a year. Would this spur me forward to actively create and finish more images? Would that help me get out of a rut and move me forward? I think it may be fun but would be an immense challenge for me. At the moment I rarely leave the house makes it a challenge or at least pushes me in a different direction.
Carrying a camera with me is not a habit that I need to adopt – my phone is always with me. I’m often mentally taking photographs – I still see the American flag, framed on 3 edges by fall leaves that I looked at for a week when I was dropping my daughter off at school but never took the photo – but I don’t take enough photos to be able to post one a day. Not yet anyway.
How about you do you carry a camera with you all the time and do ‘visual push-ups’ every day? Want to join me in the challenge? Need an accountability partner for your project? A year too long? How about a sprint? Everyday for a month? Let me know here or tag me on Instagram.
I’m currently thinking about projects that I can engage with while travel is restricted and we are locked down. I feel like a good photography project, one that will keep you engaged for a while, is one that is multi-dimensional, one that engages multiple areas of your interest.
Thomas Joshua Cooper certainly found this with his Atlantic Basin Project. The ambition of this project was to chart the Atlantic Basin from the extremity of land north, south, east, and west. An enormous undertaking and one he has been engaged with since the end of the ‘80s. Longer than he’s been married and a project older than his kids!
I find the work absolutely captivating, although I struggle to put my finger on what it is about his photographs that draws me in. The subject matter appeals to me. I enjoy being at the coast, I’m awed by the power of the ocean and can’t help but feel that the transition from land to water is a place of great possibility.
The images of Thomas Joshua Cooper for this project are abstractions with no real sense of place, not grand vistas but truly what it would feel like if you were stood at the edge of the land looking out to the sea. Often they appear to be long exposures, giving movement to the water which really gives you a sense of dynamics. There are images that provide visual breaks – the images taken during white out conditions on Antarctica or those taken during the winter solstice at the North Pole.
If you only get one book from this project get ‘The World’s Edge‘ if you want to dig deeper you could explore The Point of No Return, Eye of the Water Ojo De Agua, or True that represent images from major sections of the work. Watch the video below to hear Professor Cooper provide an entertaining and engaging description of his Atlantic Basin Project.
I must have been living under a rock to only recently have found Rachael Talibart’s seascape work. She is perhaps best know for her Sirens photos, a series of storm waves named after mythological beings. A book of the same name was published by Triplekite Books in 2018. As an aside I can’t believe I missed this book since I thought I had all of the books that Triplekite had published. I found her work through the recently published book, Tides and Tempests, that further explores her interest in storm waves but also the coast in general.
It sounds like Rachael has had a lifelong relationship with the sea having grown up on the South Coast of England, spending time as a child on the family sailboat. She describes herself as a poor swimmer and a poor sailor who is happier and safer viewing the ocean from the shore. Reading between the lines in the introductory essay to Tides and Tempests it sounds like she had lots of ‘fun’ on the sailboat as a child. These episodes really do shape your life both as an adult and as a child, either pushing you away or drawing you in. Personally I’m glad that she is drawn towards the ocean and chooses to capture the majesty, power and potential that the ocean offers. Check out more of Rachael’s work on her website here.
Also check out the videos that Rachel put together below. Scroll all the way to the bottom to hear Rachel talk about her work and her process.
Check out the mini-documentary/interview with Rachel that Sean Tucker put together below. Sean is worth a ‘Friday Inspiration’ slot of his own. Until then check out his YouTube channel here.
For me just as the beginning of summer is when we get the boat in the water, the end of summer is when we pull the boat out of the water. This weekend marked the beginning of that process as we made the trek from our mooring to the marina in Norwalk where will be hauled.
This year we were accompanied by Jay our buddy from TowBoatUS who gave us a tow to the dock. If you have a boat a BoatUS membership with on water towing is an amazing insurance policy. I highly recommend it!

domain: artistic


I attended the public consultation process for the proposed new Bus Rapid Transit system, on 17th February, at County Hall. I met some residents from Broadmeadows, who were furious at the proposed route. While we badly need a better transport system, this system is just a glorified QBC. If this goes ahead, it looks like the end of Metro, if the NTA literature is anything to go by. This whole project was suddenly sprung on the local community, without any prior consultation with residents.
View the story in the North County leader here


Voting for our 2010 Readers’ Choice Awards is continuing at a ferocious pace, with thousands of votes already counted. The Readers’ Choice Awards let you pick the year’s best from four categories—Apple’s best new digital media product, iPad/iPhone/iPod Accessory Developer, iOS Application Developer, and iOS Game Developer. The iPad and iPhone 4 are battling it out at the top of the Apple media product category, but the fourth-generation iPod touch has emerged as a dark horse and has a shot to pass them both. Apple continues to lead the Application Developer of the Year category, with Netflix, Peapod Labs, and Flipboard all making pushes of their own, while Belkin, Griffin, iHome, and Logitech are in a hotly-contested race in the Accessory Developer of the Year category. Finally, PopCap has made a recent surge in the race for Game Developer of the Year, but Rovio, EA, and Gameloft are all in the mix, as well. Write-in votes in each of the categories are significant, so if your favorite company’s not on there, write it in! Voting ends at midnight Eastern Time on October 27, and all it takes is filling out a simple four-question survey, so cast your vote today!
- MacBook Pro with Mini-LED to launch this year
- iPad Pro vs iPad Air: Shopping guide!
- Apple Watch continues to be the smartwatch leader
- Should you buy the iPad Pro or wait?
- The Anker Soundcore Motion Boom Outdoor Bluetooth Speaker falls to $79.99
- iPhone 11 submerged for 6 months gets returned to owner
- iMac Pro listed with only one configuration, set to be discontinued
- LG developing display for the foldable iPhone
- Apple Watch sales shine in 2020 Q4
- Apple to introduce tracking safety feature to prevent stalking


text:
Good evening, hope you’ve enjoyed a great weekend. I saw some family members that I haven’t seen for about a year last night at a family party which was lovely.
I thought I’d share a very cool book swap idea with you tonight as I’m planning on doing it tomorrow morning – hopefully it’ll brighten my Monday! It’s a book swap run by the Guardian and Observer newspapers and the idea is to basically choose a book that you have read and no longer need, then put on one of the newspaper’s book labels which advertise that this book is now up for grabs and leave in a public place for someone to find! If you’re interested in joining in you can download a copy of the label here. You can then post the location of your book on a flickr page and also join in on twitter if you so wish!
If I travelled to work on the tube then I’d definitely leave a book there but as I don’t I think I’ll leave mine at a train station instead. Now, which book to choose….
On that note what book would you leave for the book swap? Leave a comment and let me know – I’m always on the look out for a recommended book. The notes section on my phone is full of books that others have recommended to me and I’m always hunting for them when I visit the library (when I haven’t got a fine that is. I’m notoriously bad at getting my books back on time. Tsk)!
Have a great Monday.

domain: library


text:
Few who have experienced Montanas Glacier National Park would argue that it takes the cake for epic scenery among U.S. parks. But if global warming continues at the current rate, the result could be a glacier park sans glaciers. To celebrate its upcoming centennial and raise awareness about the chilling speed at which its namesakes are melting, the park recently co-commissioned four artists to create site-specific works on park land. Videographers then captured the works on film for integration into subsequent stage performances. Among the artists chosen for the project was choreographer Trey McIntyre, whose maverick brand of ballet has earned him growing international attention for a decade. Having created nearly 80 dances for bulwarks such as Stuttgart Ballet and New York City Ballet, McIntyre launched his own company from his home base in Boise three years ago. The Trey McIntyre Project makes its West Coast debut with "The Sun Road and Other Works." Footage of the company dancing among Glaciers majestic peaks serves as a backdrop to live ballet on the stage. Given the dancers backgrounds with some of the best ballet companies in the country, this one-night performance should offer some dancing to remember if Glaciers imperiled grandeur doesnt upstage it.
Fri., Oct. 30, 8 p.m., 2009

domain: geography


text:
PCBA Surface Mount Technology SMT Processing OEM ODM
Contact Person : Ingrid
Phone Number : +86 18126235437
WhatsApp : +8618774804503
OEM DIP Insert Processing,
ODM DIP Insert Processing,
PCBA pcb designing and manufacturing
DIP (Dual In-Line Package) insert processing is a manufacturing technique used to insert through-hole components into a printed circuit board (PCB) that has pre-drilled holes. DIP components have leads or pins that extend from the bottom and are inserted through the holes in the PCB. The DIP insert processing typically involves the following steps:
1. PCB Preparation: The PCB is prepared for DIP component insertion. This involves ensuring that the PCB has pre-drilled holes in the correct locations and sizes to accommodate the DIP component leads.
2. Component Preparation: The DIP components are prepared for insertion. This may involve straightening or aligning the component leads to ensure they can be easily inserted into the PCB holes.
3. Insertion: Each DIP component is manually or automatically inserted into the corresponding holes on the PCB. The component leads are carefully aligned with the holes and inserted until the component body rests flush against the PCB surface.
4. Securing: Once the DIP components are inserted, they are secured to the PCB to ensure they remain in place during subsequent manufacturing processes and the product's lifecycle. This can be achieved through various methods, such as soldering, crimping, or using adhesive.
5. Soldering: After the DIP components are inserted and secured, the PCB is typically subjected to a soldering process to create reliable electrical connections. This can be done through wave soldering, selective soldering, or hand soldering, depending on the production setup and requirements.
6. Inspection: Once soldering is complete, the PCB undergoes inspection to verify the quality of the solder joints and component placement. Visual inspection, automated optical inspection (AOI), or other testing methods may be employed to detect any defects, misalignments, or soldering issues.
7. Testing: The assembled PCB, with the inserted and soldered DIP components, may undergo functional or electrical testing to ensure it meets the required specifications and performs as intended.
8. Final Assembly: After testing, the PCB may proceed to final assembly, where additional components and processes are added to complete the product.
DIP insert processing is commonly used for components that cannot be mounted using surface mount technology (SMT) or for applications where through-hole components are preferred due to their robustness or specific electrical requirements.

domain: design


text:
The event did not disappoint and in just 48 hours the Palestine Area Chamber experienced a 30% increase in their membership. Not only did they increase their membership they also introduced a new investment schedule in a very exciting and effective way. With that introduction they are now able to better serve their members!
A different Chamber, a different event but eerily similar in their success story as another you just witnessed. The Palestine Area Chamber of Commerce decided to tackle the month of January by hosting a membership campaign with Your Chamber Connection. Really they wanted to start the year off with a bang, to make an impact that wouldn’t be easily forgotten. They wanted their community to understand who the Chamber was, what their mission is and add whole lot of excitement to it.
The education and energy that was stirred up here created a small town success story that all should look to.
Congratulations on your 126 new Chamber members!

domain: economics


5620 G-Star Elwood 3D Skinny Jeans
AU$ 200.00 including VAT
5620 G-Star ElwoodThe 5620 G-Star Elwood was first introduced in 1996, and is the original design from the G-Star 3D Denim approach to construction. Head Designer Pierre Morisset pioneered this method as he sought to recreate the jeans of a passing motorcyclist, which had moulded to his legs from longtime exposure to the elements.
The G-Star Elwood jeans mapped out a new way of thinking about denim when first released. This seasonal version features the same anatomical 3D construction as the original, updated in a current fit.
Super Slim 3D Fit
- Fitted waistband
- Regular rise
- Tight, narrow legs
- Zip fly
Tricia Gray Super Stretch Denim
The 5620 G-Star Elwood 3D Super Slim Jeans is cut from soft denim with a versatile gray base shade and great stretch.
- Lighter weight 9 oz denim
- Slubby texture
- Excellent stretch and recovery
- 98% cotton, 2% elastane (Lycra®)
Skillfully finished with deep fades and authentic wear marks.
40° normal process
Art. nr.: 51026-9444-071


text:
The Association of Women Lawyers (“AWL”) has commissioned a “Baseline Study on the Working Conditions of Male and Female Lawyers in Kuala Lumpur and Selangor” together with the Bar Council, SUHAKAM and the Women’s Aid Organisation (“WAO”) whereby legal and gender experts and a statistician from the University of Malaya (“UM”) have been engaged as consultants of the study.
The study aims to ascertain, among others, the working conditions of lawyers, their job satisfaction and the existence of any form of gender discrimination, if any, in the legal profession in Malaysia. The survey is to be conducted among 450 lawyers in total with almost equal representation of both male and female lawyers from various divides of the private practice groups existing in the Malaysian legal framework. As such, the Klang Valley lawyers may receive calls from UM seeking appointments with you to conduct the survey.
The respondents of the study are rest assured that strict confidentiality of all data and information shared in the survey shall be maintained at all material times. Therefore, the Project Commissioners and the Consultants seek utmost cooperation of the Members of the Bar in the Klang Valley to be part of the study if you are one of those selected randomly for the purposes of the study.
If you need further information please contact either Meera Samanther, President of the Association of Women Lawyers at email@example.com or 012-313 8850; or Dr Lai Suat Yan, Lead Consultant of the baseline study at firstname.lastname@example.org or 016-2868 543.
Contributed by Meera Samanther, President, Association of Women Lawyers

domain: law


text:
A Short Biographical Dictionary of English Literature/Buckingham, George Villiers, 2nd Duke of
Buckingham, George Villiers, 2nd Duke of (1628-1687).—Dramatist, s. of the 1st Duke, who was in 1628 assassinated by Felton. His life was full of adventure and change of fortune. The Restoration gave him back his already twice lost estates, which he again squandered by a life of wild extravagance and profligacy at Court. He was a member of the "Cabal" and intrigued against Clarendon. He wrote pamphlets, lampoons, and plays, but his chief contribution to literature was The Rehearsal, a comedy, in which he satirised the heroic drama of Dryden and others. It is believed that S. Butler had a hand in it. Dryden had his revenge in his picture of B. as Zimri in Absalom and Achitophel.


Jeffers to face Ipswich
FRANCIS Jeffers will make his Sheffield Wednesday debut against Ipswich Town on Saturday after agreeing a three-year-deal at Hillsborough.The 26-year-old striker has agreed terms of around £10,000 a week to play for the Owls after a £700,000 move from Blackburn Rovers.
By Derek Davis
FRANCIS Jeffers will make his Sheffield Wednesday debut against Ipswich Town on Saturday after agreeing a three-year-deal at Hillsborough.
The 26-year-old striker has agreed terms of around £10,000 a week to play for the Owls after a £700,000 move from Blackburn Rovers.
Jeffers turned down a move to Portman Road last month, despite being offered more than he was being paid at Ewood Park.
You may also want to watch:
The former Everton and Arsenal striker had told Ipswich Town he did not want to sign for another Championship club after the Blues revitalised his career during loan spell when he scored four goals in nine games, with a hamstring injury in the middle.
But after failing to accept personal terms with Town he held unsuccessful talks at Leicester City and Cardiff City, before settling on the Owls two days before the season starts.
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- 4 'You either deliver or you leave' - Cook's message to Town players
- 5 Paul Cook speaks about Ipswich Town takeover for first time
- 6 National Trust 'deeply saddened' at death of volunteers in Woodbridge incident
- 7 Woodbridge community 'saddened' after couple found dead by police
- 8 The first five jobs for Ipswich Town's new owners
- 9 Serious crash closes road in Bury St Edmunds near A14
- 10 Long queues at Primark from 7am in Ipswich as lockdown eases


text:
|Object type||Last Post film|
Australian War Memorial
|Place made||Australia: Australian Capital Territory, Canberra, Campbell|
|Date made||26 December 2018|
First World War, 1914-1918
Item copyright: © Australian War Memorial
This item is licensed under CC BY-NC
|Copying Provisions||Copy provided for personal non-commercial use|
The Last Post Ceremony commemorating the service of (7196) Corporal Robert Simpson Baird, 14th Australian Infantry Battalion, AIF, First World War.
The Last Post Ceremony is presented in the Commemorative area of the Australian War Memorial each day. The ceremony commemorates more than 102,000 Australians who have given their lives in war and other operations and whose names are recorded on the Roll of Honour. At each ceremony the story behind one of the names on the Roll of Honour is told. Hosted by Craig Berelle, the story for this day was on (7196) Corporal Robert Simpson Baird, 14th Australian Infantry Battalion, AIF, First World War.
7196 Corporal Robert Simpson Baird, 14th Australian Infantry Battalion, AIF
KIA 24 March 1918
Story delivered 26 December 2018
Today we remember and pay tribute to Corporal Robert Simpson Baird.
Robert Baird was born to James and Norah Baird in 1896 in Blowhard, near Ballarat in Victoria. After attending the local school, he worked as a farmer in the district.
After Australia’s involvement in the First World War was announced, Baird was keen to enlist in the Australian Imperial Force, but as he was under 21 years of age, he had to have his parents sign their consent. He joined in July 1916 and undertook a brief period of training in Australia. Before leaving for overseas service, he took home leave to visit his family a final time. During this visit, Baird contracted meningitis and had to spend some time convalescing in hospitals in Ballarat and Geelong.
Having recovered fully, Baird embarked from Melbourne in February 1917 on the transport ship Ballarat, arriving at Devonport, England, in April. After three months of training in England, he sailed to France in July.
He joined the 14th Australian Infantry Battalion, which was moving over the border from northern France into Belgium. It was the beginning of the campaign that would become known as the Third Battle of Ypres. In late September, the unit went into battle at Polygon Wood, and Baird was wounded when a shell fragment struck his head. He was evacuated to the Canadian General Hospital in Boulogne, France, where he recuperated for a month. Late in October, he returned to his battalion in Belgium.
After the Australian involvement in the Third Battle of Ypres concluded in October 1917, the 14th Battalion spent time in northern France and Belgium over the winter. Baird and the other men of the 14th undertook training behind the lines, as well as spending two weeks in the front line during January and February. At the end of February, the 14th Battalion moved into billets at Neuve Église, near the French border.
In March 1918, German forces launched what was to be their last great Western Front offensive of the war, known as the German Spring Offensive. In the north, the German army attempted to retake the ground they had lost in the fighting around Ypres. German forces shelled the town of Neuve Église regularly in order to disrupt Commonwealth supply lines. On 24 March 1918, Baird was hit by a German shell and killed instantly. He was 21 years old.
Baird was buried at Westhof Farm Cemetery in Belgium, alongside 130 Commonwealth First World War servicemen. His grieving parents had his headstone inscribed with the epitaph, “Father in thy gracious keeping, leave we now our loved one sleeping”.
Corporal Robert Simpson Baird is listed on the Roll of Honour on my right, among almost 62,000 Australians who died while serving in the First World War.
This is but one of the many stories of service and sacrifice told here at the Australian War Memorial. We now remember Corporal Robert Simpson Baird, who gave his life for us, for our freedoms, and in the hope of a better world.
Historian, Military History Section
Video of The Last Post Ceremony commemorating the service of (7196) Corporal Robert Simpson Baird, 14th Australian Infantry Battalion, AIF, First World War. (video)

domain: history


Welcome to Move Forward’s weekly news wrap-up, featuring the mobility stories you don’t want to miss. This week we’re featuring an analysis of the fastest and slowest commutes in the country’s largest cities, along with news of nation’s first autonomous shuttle system, a push for “20-minute cities” in Australia, and more.
De Blasio proposes new tax to raise funds for subway system:
New York City Mayor Bill de Blasio has proposed a new income tax on the city’s wealthiest people to help generate revenue for NYC’s ailing and overcrowded public transit system.
York Times: “London Uses Congestion Pricing. Why Doesn’t New York?” by Vikas Bajaj and Stuart A. Thompson, August 10, 2017.
How slow is your city?:
The Washington Post features an analysis of commute times and distances from leading cities across the U.S., in particular examining the impact of congestion and other factors. Cities with the slowest commute times include Boston, Washington D.C., New York, and Miami.
The Washington Post: “How Fast Can You Leave Town During Rush Hour?” by Sahil Chinoy, August 11, 2017.
Nation’s first autonomous shuttle to debut:
Arlington, Texas is launching what they believe is the nation’s first autonomous public shuttle network. Starting on August 26, Milo, short for Mile Zero, will bring fans from remote parking lots to games at the AT&T Stadium and Globe Life Park.
Government Technology: “Arlington, Texas, to Launch ‘Nation’s First Autonomous Public Shuttle Network’” by Bill Hanna, August 11, 2017.
Cities look to “future proof” their streets:
Many experts have predicted that within the next 15 to 30 years, the digital world will influence transportation systems– from high-speed tunnels, to drones, and most importantly, autonomous vehicles. The implications of these innovations could extend deeply into the fabric of urban communities, and now public leaders are faced with the dilemma of “future proofing” their communities in preparation for disruption.
Governing: “Transportation and the Challenge of Future-Proofing Our Cities” by Bob Graves, August 14, 2017.
In Australia, there is a significant push in government for the adoption of the “20-minute city,” which seeks to set a maximum trip duration of 20 minutes for all modes of transportation. New research in Melbourne has found that so far, only 3% of public transit trips take less than 20 minutes, compared to 60% of trips by car.
Crickey: ”Can public transport define the 20-minute city?” by Alan Davies, August 16, 2017.
Fiat Chrysler joins BMW autonomous vehicle alliance:
A new partnership between BMW, Intel, Mobileye, and Fiat Chrysler will aim to produce an industry-wide autonomous car technology which other carmakers could adopt. This alliance is part of a new trend in the automotive world as industry leaders seek ways to share the high costs of developing autonomous technologies.
Reuters: “Fiat Chrysler joins BMW-Intel self-driving car alliance” by Alexandria Sage and Edward Taylor, August 16, 2017.
Uber considers selling U.S. car leasing business:
A report released earlier this week informed Uber’s executives that losses at Xchange Leasing were $9,000 per car on average. These numbers were far above the predicted estimates of around $500 per car, prompting conversations of selling the company.
Automotive News: “Uber considering sale of U.S. car-leasing business, report says” by Parikshit Mishra, August 16, 2017.
Industry leader discusses rapidly changing transit space:
Harriet Tregoning, former leading official at U.S. Department of Housing and Urban Development, discusses how cities should be planning and preparing their transportation infrastructure for the future.
Government Technology: “How Cities Should Plan and Prepare Their Transportation Infrastructure for the Future” by Bob Graves, August 16, 2017.
Please note that this article expresses the opinions of the author and does not reflect the views of Move Forward.


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domain: sociology


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Book review of great debut novel, WANDERER SCORNED by Natasha Woodcraft
As an author, it is a real honour to be asked to read and review another author's work. So many authors have helped me on my writing journey, it only feels right to do the same. This is the first 'Blog Tour' that I have taken part in and this book is a definite MUST READ.
The debut novel from Natasha Woodcraft, called ‘Wanderer Scorned’ is an adaptation of the story of Cain and Abel from the Bible. If you aren’t aware, Cain killed his brother, Abel. So, just like an episode of Columbo, the end is already known and will require a high standard of storytelling, if it is going to grip your attention. Natasha Woodcraft achieved this beyond my expectations.
Why do I say beyond? Well, it is of course Biblical Fiction, which I used to say is my least favourite genre of writing, but I am rapidly changing my thinking on this. This story is extremely well researched, skilfully composed and clearly directed by the Holy Spirit. On top of all of that, it made me see things and consider things in a new light. Not that it was a distraction from the Biblical facts, indeed, it was the opposite, it brought an already known story alive.
The names of the main characters have been changed to Kayin and Habel, which helps to remind you that it is a work of fiction. The theme of murder is popular in stories, perhaps because we all have the potential in us. Really though, this story is all about relationships, family and our human frailty. Written in the first person, the author places you into Kayin’s head. You hear his thoughts and feelings, allowing you to realise that he is not a monster.
We are all capable of pride, envy and jealousy, with the possibility for it to overpower and consume us. This story follows Kayin’s devolution and dysfunctionality. In the world today, we all too often hear of dysfunctional families, yet this is not a modern issue. Cain was the first offspring of Adam and Eve, who were punished with banishment from the garden. When we look at Cain or Kayin, we find that the apple doesn’t fall far from the tree (no pun intended).
The main story is sandwiched between a prologue and epilogue, which is set at a party several generations after the killing. Kayin arrives during the prologue to tell his story. The epilogue leaves you knowing that there is more to come, more stories and more books. I’m not sure how many books will be in the Wanderer series, but if they are as good as this one, I want to read them all and I highly recommend this outstanding debut. The book has a well-deserved 5 stars from me.
Brendan Conboy is the author of 9 published books including his fascinating autobiography, The Golden Thread. In 1986, Brendan invited Jesus into his life and God blessed him with the gift of rhyming words. He used that gift as a Christian Rap artist for 25 years and has written 3 poetry books including the entire Book of Psalms in Rhyme. He has 3 published novels - Issues, Invasion of the Mimics and Legacy of the Mimics. Visit Brendan's website HERE.


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Ancient Laodicea, once a thriving city, now lies in ruins, awaiting a more thorough excavation than it has so far received. Overshadowed by the more spectacular nearby site of Hierapolis (Pamukkale), Laodicea receives the occasional busload of tourists who stop to view the remains of this city that the book of Revelation imagined as having boasted, “I am rich, I have prospered, and I need nothing” (3:17). Laodicea is south of the modern village of Goncalï and north of the village of Eskihisar. The site is located on a plateau between two small rivers that are tributaries of the Lycus River. The Asopus River runs along the western part of the ancient city, while the Caprus River runs along the east. To visit the site, take the road from Denizli that leads to Pamukkale. Two different roads from the Denizli-Pamukkale highway lead to Laodicea, both of which are on the left and marked with a sign indicating the way to Laodicea. Laodicea is situated 10 miles from Colossae and 6 miles from Hierapolis. This area was a part of the region of Phrygia, although it was sometimes considered a part of Lydia or Caria. Pliny the Elder claims that Laodicea was built on the site of an earlier settlement known as Diospolis and later as Rhoas (Natural History 5.105). Because of its location near the Lycus River, the city was known as Laodicea ad Lycum in order to differentiate it from several other cities named Laodicea. Of particular importance to the commercial success of the city was its position at the junction of two roads—one that ran from the Aegean coast near Ephesus through the Meander River valley and on to the Euphrates, and another that ran from Pergamum to Sardis and then to Perga and Attalia (modern Antalya). Antiochus II, the Seleucid king (r. 261–246 B.C.E.), founded the city during the middle of the 3rd century B.C.E. He named the city in honor of his wife Laodice, whom he later divorced. After the Romans, with the aid of the Pergamene kingdom, defeated Antiochus III at Magnesia in 189 B.C.E., Laodicea came under the control of Pergamum.
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Affordable therapeutics are vitally needed for humans worldwide. Plant-based production of recombinant proteins can potentially enhance, back-up, or even substitute for the manufacturing capacity of the conventional, fermenter-based technologies. We plastome-engineered a tobacco cultivar to express high levels of two “plantakines” — recombinant human cytokines, interleukins IL-37b and IL-38, and confirmed their native conformation and folding. Assessment of their biological functionality was performed ex vivo by analyzing the effects exerted by the plantakines on levels of 11 cytokines secreted from human peripheral blood mononuclear cells (PBMCs) challenged with an inflammatory agent. Application of the plant-produced IL-37b and IL-38 in PBMCs stimulated with Lipopolysaccharide or Phytohaemagglutinin resulted in significant, and in particular cases—dose-dependent modulation of pro-inflammatory cytokines secretion, showing attenuation in two-thirds of significant level modulations observed. Plantakine treatments that increased inflammatory responses were associated with the higher dosage. Our results demonstrate feasibility of manufacturing functional recombinant human proteins using scalable, cost-effective and eco-friendly plant-based bioreactors.
Plants make a lot of sense as production platforms for all kinds of biologics. Photosynthetic capacity allowing autotrophic growth renders plants the most energy-efficient and cost-effective platform for manufacturing of various recombinant proteins, secondary metabolites and other assorted small molecules, as plants require only three abundantly available raw input ingredients for biosynthesis—carbon dioxide, water and sunlight. Hence, the initial part of the manufacturing process, the "upstream production" that generates the biomass accumulating the desired product ensues significant costs savings, eliminating the need for construction, maintenance and operation of fermenter facilities1,2,3. Benefits to “downstream production” steps of the process, where the desired product is extracted and purified are also recognized for plant-based systems, with some of the bottlenecks being addressed in recent studies4,5,6. Additional advantages of exploiting plants as single-use, clean and biodegradable bioreactors for production of recombinant proteins include inherent safety due to inability of mammalian pathogens to propagate in plant tissue and virtually unlimited scalability of plant-based production7,8.
Since the emergence of the first reports of successful genetic transformation of plants and the expression of recombinant heterologous proteins of human origin in transgenic plants, tremendous technological advances were achieved in the “molecular pharming” field, with the first FDA-approved pharmaceutical for human use in 2012, taliglucerase alfa, produced in carrot cells9. Today several biopharmaceuticals on the market are sourced from plants and a few biotechnology companies around the world use plant-based production platforms in their manufacturing processes3,6,10. Plant-based bioreactors could facilitate making more affordable many biologic drugs in use today and provide a source of therapeutics supplied locally, which can be very beneficial in the context of developing nations, or when global supply chains are disrupted6,11.
Among the methodologies used for plant-based recombinant protein manufacturing, plastome-engineered plants possess several advantageous features as a platform, simply generating extraction-ready biomass from seed. Plastome-engineered plants can express and accumulate very high yields of the desirable product and, thus, can represent the most cost-effective production route12,13,14,15. We set to demonstrate the feasibility of plastome-engineered plant bioreactor platform for production of biologically active recombinant human cytokines. Based on our preliminary screens searching for valuable proteins with a potential for prolific expression in plastids, we engineered the plastome of a low-alkaloid tobacco cultivar to produce “bioreactor lines” expressing mature forms of two “plantakines”—human interleukins IL-37 (isoform b, IL-37b) and IL-38, both characterized as anti-inflammatory cytokines16,17. IL-37b and IL-38 belong to the IL-1 family of 11 interleukins, 7 of which are pro-inflammatory18. Both IL-37b and IL-38 function in regulation/mitigation of human inflammatory responses; a plethora of studies demonstrated central involvement for IL-37b and IL-38 in immunity and disease and, therefore, as potential candidates for development as therapeutic agents19,20. The created plastome-engineered bioreactor lines produced up to ~ 1 g of the recombinant protein per 1 kg of fresh leaf biomass. After confirmation of their correct folding via protein-specific ELISAs, we assessed the biological activity of the plant-produced IL-37b and IL-38 in ex vivo experiments by monitoring the response to inflammatory agents (IAs) in freshly isolated cultured human peripheral blood mononuclear cells (PBMCs), manifested in the levels of secreted inflammatory cytokines.
PBMCs are the central and crucial components of the immune system that brings forth a response to intruder pathogens, as well as identifies and fights own body cells that have undergone malignant transformation (cancer). PBMCs are an assorted mixture of highly specialized immune cells, PBMCs population is comprised of a multitude of immune cell types including lymphocytes (~ 85%), monocytes (~ 15%) and dendritic cells (< 1%)21. In vitro and ex vivo human PBMCs studies are ubiquitous in cell biology and immunology research and an important biotechnological tool in developing new therapeutics and diagnostics22,23,24,25,26. We hypothesized that by monitoring the inflammatory response in IA-stimulated PBMCs we could study the effects of the plantakines (and their active concentrations) exerted on the levels of specific inflammatory markers.
We report significant modulation of inflammation responses from PBMCs stimulated with different IAs as a result of treatments with the plant-produced IL-37b and IL-38. We observed attenuation of levels of several secreted inflammatory cytokines, generally consistent with the previous reports characterizing the biological activity of IL-37b and IL-38 as anti-inflammatory. Both plantakines exerted dose-dependent modulations of PBMCs responses, leading at different concentrations to either inhibition or enhancement of secretion of some of the inflammatory markers monitored. In addition, different IAs brought about different magnitude of inflammatory responses reflected in levels of cytokines secreted from the stimulated PBMCs, confirming a similar experimental outcome reported recently. Thus, our study validates applicability of the plant-based production platform for cost-efficient and eco-friendly manufacturing of functional recombinant human cytokines in large quantities.
Monomers, dimers and multimers of IL-37b and IL-38 accumulate in engineered chloroplasts
We engineered the plastome transformation constructs to produce IL-37b and IL-38 as mature peptides (V46–D218 for IL-37b, C2–W152 for IL-38), optimizing the expression by selecting suitable cis-acting regulatory genetic elements and using plastid-preferable codons (data not shown). Screening for prolific producer lines of IL-37b and IL-38 identified the best configurations of plastid expression cassettes by examining their crude leaf tissue extracts with Western blots (Fig. 1a). Two bioreactor lines were selected and grown in greenhouse to maturity, expressing the recombinant human IL-37b and IL-38 at ~ 1 g and 0.75 g, respectively, per 1 kg of fresh leaf tissue. Interestingly, prevalent amounts of both plantakines were found to accumulate in older leaves, demonstrating significant stability of these recombinant proteins in the chloroplasts (Fig. 1b). We observed large amounts of the monomeric forms, as well as the dimerized and multimerized forms of the cytokines IL-37b and IL-38 in the crude leaf extracts and in samples after purification; the dimers (and higher molecular weight multimers) were very stable and detectable in SDS-PAGE analyses gels even after harsh denaturing conditions of the sample preparation. That observation was in stark contrast to the bacteria-produced recombinant IL-37b and IL-38 counterparts available commercially, that predominantly presented the monomeric forms of the cytokines when used as controls in Western blot experiments using specific antibodies (Fig. 1c,d). Placement of the HIS-tag at the N-terminal had no effect on the formation of dimers/multimers for both expressed cytokines (data not shown).
Plantakines bioactivity assessment—experimental design
The bioactivity of our plantakines IL-37b and IL-38 was assessed by monitoring the secretion of 11 cytokines, generally regarded as inflammatory markers, from PBMCs stimulated with an IA. Freshly isolated human PBMCs were subjected to various treatments—combinations of the IAs with the plantakines at different concentrations. Two different model IAs were used, either the bacterial lipopolysaccharides (LPS) or a lectin from Phaseolus vulgaris (phytohaemagglutinin, PHA); each of the IAs was applied onto cells separately, each IA was applied at two concentrations: LPS at 150 and 300 pg/mL; PHA at 5 and 10 µg/mL. Each IA at each concentration was applied in combination with one of the two plantakines, each of them at three different concentrations: 1, 10 and 100 ng/mL of the monomeric forms present in the purified extracts. Also included were treatments comprised of either IA at their lower concentrations, in combination with both plantakines at 10 ng/mL concentration in order to assess possible synergistic effects. Cells with only IAs applied represented the reference (positive controls for each concentration), cells without any treatment represented the basal level (negative control). The levels of eleven different pro-inflammatory cytokines secreted into the medium from the PBMCs—GM-SCF, IFNγ, TNFα, IL-1α, IL-1β, IL-6, IL-8, IL-22, IL12, IL-17 and IL-10 were compared between the treatments and the controls. We applied generalized estimating equation (GEE) model for the statistical data analysis. This statistical approach allows for nested observations and was used to test the effects of the plantakines IL-37b and IL-38 and their dosage in the context of IA-stimulated PBMCs inflammatory responses.
Different IAs bring about different magnitude of inflammatory responses from PBMCs
In order to validate the obtained data, as well as to gain insights into the quantitative and qualitative differences in the PBMCs’ inflammatory responses between the two IAs tested, we first compared the mean values for each monitored secreted cytokine elicited by either LPS or PHA. The magnitude of the general responses from the stimulated PBMCs, manifested in levels of the secreted pro-inflammatory cytokines was found significantly different between the two IAs. PHA elicited stronger response in 7 out of 11 pro-inflammatory cytokines monitored; namely, IL-17, IFNγ, TNFα, IL-12, IL-22, IL-10 and IL-6 displayed, respectively, 2930%, 1240%, 55.7%, 118%, 21.5%, 86.4% and 10.1% higher levels, compared with the LPS-elicited levels (p < 0.001—p < 0.05, Fig. 2). The levels of GM-CSF, IL-8, IL-1α and IL-1β showed no statistically significant difference between the IAs in our experiments. Further dissection of the data focusing on the differences in levels of the inflammatory markers between the lower and higher doses of each IA revealed that the higher dose of PHA elicited higher levels of IFNγ (+ 249%), IL-1α (+ 30%), IL-1β (+ 35%), IL-12 (+ 65%), IL-17 (+ 54%), TNFα (+ 49%) and IL-10 (+ 51%), compared with the lower dose. Higher dose of LPS brought about higher secreted levels of GM-CSF (+ 5%), IL-1α (+ 17%), IL-1β (+ 20%) and IL-10 (+ 26%), while secretion of IL-17 decreased (− 12%).
Plant-produced IL-37b and IL-38 modulate inflammatory responses from IA-stimulated PBMCs
To gain insight into the bioactivity of the plantakines IL-37b and IL-38 exerted on IA-stimulated PBMCs we compiled the data generated from treatments with observed modulatory effects on secreted inflammatory markers. GEE analysis was performed four times, for each combination of the IA and its concentration, for each secreted cytokine monitored, generating statistically significant (p < 0.05) level modulations displayed in 118 treatments out of the total 286 treatment combinations assessed. Plantakines exerted statistically significant modulatory effects on the levels of secreted inflammatory cytokines in 67 and 51 treatments that occurred in LPS- and PHA-stimulated PBMCs, respectively (Table 1). Collectively, treatments with plantakines IL-37b and IL-38 resulted in more profound anti-inflammatory activity in LPS-stimulated PBMCs rather than PBMCs stimulated with PHA, as only 10 treatments resulted in increased secretion of inflammatory cytokines in LPS-stimulated PBMCs, while decreased secretion was observed in 57 treatments. In contrast, secretion of inflammatory markers in PHA-stimulated PBMCs was suppressed in 17 treatments with plantakines and increased in 34. Notably, all the treatments with the simultaneous application of both IL-37b and IL-38 brought about increases in secretion of inflammatory cytokines under stimulations with either IA, while separate applications of the plant-produced IL-37b or IL-38 suppressed secretion of inflammatory cytokines in 44 and 30 treatments, and increased it in 12 and 22, respectively. Fewer treatments with plantakines caused suppression of inflammatory cytokines secretion and the numbers of treatments where inflammatory cytokines secretion increased grew in association with a higher concentration of either IA used to stimulate the PBMCs (Table 1).
We further analysed the changes in levels of the secreted inflammatory cytokines from the stimulated PBMCs resulting from treatments with different doses of the plantakines IL-37b and IL-38. For each inflammatory marker monitored, the outcomes of the treatments were calculated as percentages of secretion modulation with its probability value in comparison with the positive controls at the corresponding IAs concentrations (Fig. 3; Table SI, Figure S1 in Supplementary Materials). The modulatory effects of the plantakines IL-37b and IL-38 could be observed on the levels of most of the monitored secreted inflammatory cytokines elicited with either LPS or PHA, showing attenuation in 63% of the statistically significant modulations. IL-37b attenuated levels of IFNγ, IL-1α, IL-1β, IL-22, IL-17 and TNFα in LPS-stimulated PBMCs, the effect could be seen at all the concentrations examined, levels of IFNγ and IL-22 were also reduced by IL-37b in PHA-stimulated PBMCs (Fig. 3; Table SI). Increases in secretion of the inflammatory markers were mostly observed after treatments with higher concentrations of plantakines, whereas 66% of the treatments where secretion increases occurred were associated with the highest dose of plantakines, followed by 28% associated with the intermediate dose and 6% linked to the lowest dose. Unexpectedly, IL-37b at all 3 concentrations brought about an increase in IL-17 secreted from PBMCs stimulated with PHA at 10 µg/mL, similar increases were observed for IL-1α and GM-CSF levels with IL-37b at 100 ng/mL. Modulation of GM-CSF levels by both IL-37b and IL-38 displayed dose-dependent character: at low concentrations (1 ng/mL) both plantakines attenuated GM-CSF levels by more than 50% in PBMCs stimulated with 150 pg/mL LPS, while 100 ng/mL plantakines concentration increased the levels of GM-CSF, those increases observed more profoundly at LPS 300 pg/mL concentration (Fig. 3). Both plantakines boosted GM-CSF in PHA-stimulated PBMCs: at higher concentrations (100 ng/mL) IL-37b brought about 155.9% and 127.8% increases in GM-CSF levels at 5 µg/mL and 10 µg/mL PHA stimulation, respectively, and IL-38 showed 380.5% and 326.6%, p < 0.001. Interesting, a combination of both plantakines, each at concentration 10 ng/mL exerted a 228.6% (p < 0.001) increase in secreted GM-CSF levels, pointing out a possible synergistic effect from the simultaneous application, since when applied separately on PBMCs with the same 5 µg/mL PHA stimulation, plantakines IL-37b and IL-38 modulated GM-CSF levels to increase 50.5% and 103.3%, respectively (p < 0.001). Simultaneous applications of both plantakines resulted in increased secretion of several pro-inflammatory cytokines from PHA-stimulated PBMCs, IL-1α, IL-1β, IL-12, IL-17, TNFα and IL-10 displayed, respectively, 35.0%, 35.0%, 134.5%, 43.3%, 61.6%, and 43.3% increased levels (p < 0.001—p < 0.05). Notably, only insignificant modulation of IL-6 and IL-8 levels was observed, yet, when plantakines IL-37b and IL-38 were applied at the lowest concentration (1 ng/mL), statistically significant attenuation (− 9.5% for IL-6 elicited at 150 pg/mL LPS, p = 0.012, and − 28.5% for IL-8 elicited at 300 pg/mL LPS, p = 0.032) was detected, aligned with the anti-inflammatory functions expected from IL-37b and IL-38 (Table SI).
In the present study we engineered the tobacco plant plastome to produce green bioreactors capable of manufacturing profuse amounts of two functional recombinant human cytokines, IL-37b and IL-38, mainly known as anti-inflammatory modulators of immune responses16,28. To our best knowledge, this is the first report describing such a prolific expression and production of both recombinant human cytokines in their active forms in plants, a previous successful attempt to produce IL-37b in tobacco via nuclear genome transformation reported much lower yields, while there are no reports on IL-38 production in plants hitherto29. Very interesting is the fact that the penultimate amino acid (the second amino acid in the peptide chain after the initiating Methionine) of the IL-38 peptide is Cysteine, which was underlined, along with Histidine, as the strongest instability-conferring penultimate amino acid for protein expression and accumulation in plastids, leading the researchers to propose existence of an N-terminus-dependent protein degradation pathway in plastids30. Our bioreactor lines produced IL-38 peptide with the penultimate Cysteine at 8–10% of the total soluble protein in the leaf tissue, contradicting the proposed model. Further, the same transformation construct expressing an IL-38 peptide variant with an added penultimate Serine, which was reported as a stabilizing penultimate amino acid, reached similar levels of IL-38 accumulation (data not shown), suggesting that the proposed N-terminus-dependent protein degradation pathway in chloroplasts is either limited in its processing capabilities, or involves additional unknown regulatory factors that specifically direct degradation of select proteins30.
The accumulated dimerized (and multimerized) forms of the plantakines IL-37b and IL-38 in the crude leaf extracts and in purified samples constituted ~ 50% of the entire recombinant protein yields, contrasting the results of successful recombinant production studies of IL-37b and IL-38 proteins in bacteria, which never reported dimerization of the purified cytokines in their SDS-PAGE analyses, even when the purified IL-37b was concentrated by ultrafiltration31,32,33. This observation suggests that the chloroplast stroma compartment, accumulating the synthesized recombinant proteins, provides a beneficial milieu of internal conditions/chaperones/scaffolds assisting the folding of these cytokines and promoting their further dimerization and multimerization. It is also reasonable to assume the remarkable stability of those dimer/multimeric forms of the cytokines in plastids, since the highest levels of accumulation were observed in older leaves. Bacteria-produced IL-37b was shown to form dimers at nanomolar concentrations and tetramers at higher concentrations, which greatly diminished its bioactivity, suggesting a mechanism of activity regulation through monomer/dimer equilibrium and leading to an engineered monomeric IL-37b variants with much stronger biological activity34,35. Those monomeric variants, however, along with the natural mature recombinant IL-37b peptide showed appearance of minor bands that corresponded to the dimer size in SDS-PAGE analyses and further investigation of these protein structures is needed34. Also, intriguing is the question whether the formation of dimers/multimers contributes to the overall stability of those cytokines against proteolysis, thus enabling the proposed mechanism of self-regulation and in situ preservation in a stable inactive form in the intercellular space, where the local IL-37b concentration reached the dimerization constant values34. A mechanism of bioactivity regulation through the dimer formation, similar to that of IL-37b, was proposed for IL-38 in a recent review28; however, no scientific reports are available addressing this subject. Further structural studies will answer the question whether IL-38 can be engineered into a stable, bioactive monomer, similarly to IL-37b.
Bodily inflammatory processes are a part of innate immune responses, promoted by pro-inflammatory cytokines released from the cells of the immune system as a reaction to the presence of an inflammatory agent or stimuli. Secreted levels of 11 well-characterized inflammatory cytokines, namely GM-SCF, IFNγ, TNFα, IL-1α, IL-1β, IL-6, IL-8, IL-22, IL12, IL-17 and IL-10 were monitored in our experiments with freshly isolated PBMCs subjected to treatments with two different IAs in combination with two plant-produced anti-inflammatory cytokines IL-37b and IL-38 at different concentrations. This experimental setup allowed for focusing on the biological effects exerted by the plantakines IL-37b and IL-38 via a direct comparison of the levels of pro-inflammatory cytokines secreted from IAs-stimulated PBMCs with or without the plantakines treatments at corresponding concentrations (Tables 1, SI).
Although PHA and LPS bind to completely different sets of receptors, both IAs seem to trigger varying signal transduction cascades, particularly leading to the activation of NF-κB, which plays an essential role in regulating the expression of genes linked to innate immunity and inflammatory responses36,37. Seven out of 11 inflammatory cytokines monitored in our study displayed significantly higher secretion from PBMCs stimulated with PHA rather than LPS, IL-17 secretion varied more than 30-fold (Fig. 1). Remarkably, our findings strongly corroborate a recent report stating that after PHA-stimulation human PBMCs secreted significantly higher levels of inflammatory cytokines, compared to stimulation with LPS, which apparently failed to notably induce IL-17, TNFα and IL-1238.
The modulatory effects exerted on levels of the inflammatory markers secreted from IA-stimulated PBMCs confirmed the biological activity of plant-produced IL-37b and IL-38. Statistically significant modulations occurred in both LPS- and PHA-stimulated PBMCs due to treatments with the plantakines (Table 1). For quenching inflammation, treatments with plantakines appeared to be more effective in LPS-stimulated PBMCs, where 85% of all treatments resulted in attenuations of the levels of secreted inflammatory markers, while only 30% of treatments attenuated inflammatory markers in PHA-stimulated PBMCs, implying varying efficacy and specificity of the anti-inflammatory action under different stimuli. Attenuated, rather than increased secretion of inflammatory cytokines occurred 3.5 and 1.7 times more frequently in treatments with IL-37b and IL-38, respectively, in accord with the proposed role for IL-37b as a primary and fundamental inhibitor of inflammation16,39. Treatments combining both plantakines, however, resulted in increased secretion of inflammatory markers under either IA; also, with higher concentrations of either IA applied for PBMCs stimulation, treatments with plantakines suppressive of secretion of inflammatory cytokines became scarcer, while more treatments caused increased inflammatory secretion (Table 1). These observed phenomena are inexplicable at this point and require further scientific exploration.
Levels of the inflammatory markers secreted from the stimulated PBMCs displayed distinct and varying patterns of modulation following application of treatments with the plant-produced IL-37b and IL-38 (Fig. 3; Table SI). The number of treatments that triggered an increased, rather than attenuated secretion, was higher only in 2 among the 11 inflammatory cytokines monitored, namely GM-CSF and IL-12, indicating general anti-inflammatory effects exerted by the treatments with plantakines in IA-stimulated PBMCs. Notable were the differences in the magnitude and the scope of the modulation, reflected in the outcomes of the treatments being either an attenuation or an increase of the inflammatory secretion levels: an attenuation of the secretion was the outcome of 74 treatments, averaging − 28% level reduction, while increases in secreted levels of inflammatory cytokines, observed in 44 treatments, displayed 79% on average. Among the 11 cytokines monitored, only the levels of IFNγ exhibited consistent attenuation from treatments with either plantakine in PBMCs stimulated with either IA, displaying also the strongest attenuation observed in our experiments (− 63.9%, p < 0.001), exerted by application of 1 ng/mL IL-38 in PBMCs stimulated with 150 pg/mL LPS. In contrast, GM-CSF levels were 3 times more frequently increased, rather than attenuated by treatments with the plantakines, with the strongest increase reaching 380.5%, p < 0.001, upon application of 100 ng/mL IL-38 in PBMCs stimulated with 5 µg/mL PHA (Table SI). Strikingly, the plant-produced IL-37b and IL-38 both exerted dose-dependent regulation of GM-CSF secreted levels, bringing about attenuation at low concentrations, while causing increases at high concentrations. Although both IL-37b and IL-38 are generally characterized as anti-inflammatory cytokines active in quenching inflammation17,40, studies have reported that recombinant unprocessed IL-38 could increase inflammatory cytokine IL-6 production in human macrophages in response to LPS or IL-1β stimuli41,42. In addition, IL-37b was reported to increase TNFα production in higher concentrations and Candida-induced IL-17 production was reportedly blocked by low concentrations of IL-38, while higher doses of IL-38 induced more IL-17 production, a pattern which resembled IL-37b bioactivity41,43.
Both IFNγ and GM-CSF are crucial cytokines for activation/differentiation of myeloid cell populations44,45 and their nuanced regulation by the plant-produced IL-37b and IL-38 may serve as a primer for future studies to discern novel patterns in PBMCs inflammatory responses. Interesting also to note that statistically significant attenuation of IL-6 and IL-8, two profound inflammation markers monitored in our study46,47 was only detected upon applications of low concentrations of the plantakines, aligned with the anti-inflammatory functions expected from IL-37b and IL-38.
In conclusion, we developed plastome-engineered, low-alkaloid tobacco bioreactor lines for cost-efficient and prolific production of two functional human cytokines with profound anti-inflammatory properties, IL-37b and IL-38, which are underlined as prospective therapeutic agents. Our explorative study demonstrated that the plantakines exerted significant modulation of levels of secreted cytokines involved in inflammatory responses monitored in IA-stimulated PBMCs, indicating a dose-dependent mode of action and general attenuation of several secreted inflammation markers. Enhancement of several pro-inflammatory cytokines, associated with higher concentrations of the plantakines applied in treatments was also observed, revealing novel patterns of inflammation regulation by IL-37b and IL-38. Different magnitude of responses from PBMCs were seen in levels of secreted cytokines elicited by different IAs, where PHA elicited stronger response than LPS in levels of most secreted cytokines monitored. Cumulatively, our results demonstrate feasibility of producing functional human recombinant cytokines in plants and further promote the accelerated adoption of plant-based manufacturing of various recombinant proteins by biotechnology industries.
Plastome-engineered bioreactor lines expressing recombinant human IL-37b and IL-38 (UniProt identifiers Q9NZH6 and Q8WWZ1, respectively) in their mature forms (V46–D218 for IL-37b, C2–W152 for IL-38), each bearing a C-terminal hepta-HIS-tag, were produced by Igor Kolotilin for Solar Grants Biotechnology Inc.
All permissions for including the plant material/samples were obtained from Solar Grants Biotechnology Inc. All experimental research work in this study, including the collection of plant material, comply with relevant institutional, national, and international guidelines and legislation.
Recombinant protein extraction and purification
Total soluble proteins from fresh leaf tissue of the bioreactor lines were extracted as described48. In short, flash-frozen leaf tissue was milled into powder with pestle and mortar and then 5 volumes of the extraction buffer (1XPBS, pH = 7.4) was added, complemented with 10 µg/mL leupeptin 2 mM PMSF and 2% PVPP. After filtration through Whatman #1 paper, the extract was centrifuged twice for 15 min at 13,000× g in 4 °C. The recombinant proteins were purified from the cleared extract utilizing the C-terminal His-tag and the immobilized metal ion-affinity chromatography (Cytiva Life Sciences™ His SpinTrap™, Cat. No. 28932171), dialysed against 1XPBS, pH = 7.4 and filtered through 0.22 µm (EMD Millipore, Cat. No. SLGV004SL) to obtain sterile solutions.
SDS-PAGE and Western Blots
SDS-PAGE and Western Blots experiments were performed as described previously48. Briefly, the clarified extract was mixed in a proper proportion with 5XSample Buffer (50 mM Tris, pH 8.0, 1% SDS, 20 mM DTT, glycerol) and the mixture was boiled at 99 °C for 30 min, after which was cooled down to room temperature and loaded into the gel wells. Anti-His-tag antibodies (GenScript, Cat. No. A00186-100), as well as the monoclonal antibodies against human IL-37b and IL-38 (MyBioSource Inc., Cat. No. MBS 7600509 and R&D Systems, Cat. No. DY9110-05, respectively) were used to detect the blotted proteins according to manufacturers’ recommendations; bacteria-produced recombinant human IL-37b (R&D Systems, Cat. No. 7585-IL-025) and IL-38 MyBioSource, Cat. No. MBS635478) were used as the positive controls. Western blots membranes were visualized using the enhanced chemiluminescence (ECL) detection kit (GE Healthcare, Cat. No. RPN2232) and imaged with the DNR Bio-Imaging System MicroChemi (RANCOM A/S, Birkerød, Denmark). Densitometry was performed using the TotalLab TL 100 software (Nonlinear Dynamics, Durham, NC).
For confirmation of the native folding and quantitation analyses, protein-specific ELISA experiments with the plantakines IL-37b (Invitrogen™, Cat. No. LS885210322) and IL-38 (R&D Systems Inc., Cat. No. DY9110-05) were performed according to manufacturers’ recommendations. BioTek Instruments (VT, USA) Epoch Microplate Spectrophotometer was used to acquire numerical ELISA data.
Experiments with PBMCs—general design and multiplex cytokine analysis
Sample collection, experimental protocols and all methods were performed in accordance with relevant guidelines and regulations. After isolation, the PBMCs from each donor separately were counted, plated in equal numbers per well and stimulated for 24 h with the applied treatments. Control wells on the plate contained media only (as basal level controls), and LPS and PHA (as positive controls at their corresponding concentrations). After the 24 h treatments stimulation the PBMCs’ culture supernatants were used in the multiplex Luminex platform-assisted analysis of the concentrations of the 11 secreted pro-inflammatory cytokines.
The whole blood from 5 random human donors was collected in ACD Vacutainer tubes and immediately processed for isolation of PBMCs by gradient density centrifugation using Lympholyte. The freshly isolated PBMCs from each donor separately were cultured in a volume of 200 µL at a concentration of 1.25 × 106 cells/mL in 96-well plates (~ 250,000 cells/well) in an incubator set at 37 °C, 5% CO2 and > 80% humidity. PBMCs were treated with one of two IAs at 2 concentrations each (LPS at 150 and 300 pg/mL; PHA at 5 and 10 µg/mL) in combination with two test items (plant-produced IL-37b or IL-38) at three concentrations each (1, 10 and 100 ng/mL), based on the monomeric form amounts estimated with densitometry (ImageJ). Each treatment was tested in triplicates. Control wells contained media only (negative control, basal level of detection) and the examined IAs at both tested concentrations as the positive controls. The PBMCs were pre-incubated with the test items41, stimulated with the IAs and incubated for 24 h after the stimulation for analysis of the levels of the secreted cytokines that were determined in the culture supernatant using a multiplex immunoassay (MAGPix®, Luminex), analytes were selected from the Milliplex panel HCYTOMAG-60K, sensitivity range of 3.2–10,000 pg/mL. All parameters of the Milliplex panel cytokines analysis using Luminex platform were validated (Millipore-Sigma).
Non-linear logarithm transformation was performed to address non-normality of distribution of Luminex-derived numeric data. Generalized estimating equation (GEE) method was used in the analysis of nested (correlated) structure of the data. Secreted inflammatory cytokine values were used as dependent variables and test item (plantakines) with dosage as independent predictors (factors). Separate analyses were conducted for each of the 11 monitored cytokines and each inflammatory agent/concentration combination. Analysis was performed using SPSS software version 27 using the level of significance 0.05 (p values < 0.05 are reported as statistically significant).
All data generated and analyzed during this study are included in this published article and its Supplementary Information files. Enquiries regarding availability of the generated plant bioreactor lines (seeds) should be sent to email@example.com.
Granulocyte macrophage-colony stimulating factor
Peripheral blood mononuclear cells
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We are thankful for expert advice from Dr. Idriss Saiah, Dr. Gurpreet Kaur Singh and Dr. Farideh Nourian (Immuni T Corp. Montreal. Quebec, Canada) in the design of the experiments with PBMCs and to Anton Svendrovski, MBA, M.Sc. (UZIK Consulting Inc.), for the proficient help with the statistical analyses. Our sincere appreciation goes to Mr. Felipe Campusano, B.Sc., B.Pharm. (Solar Grants Biotechnology Inc.) for his support and guidance throughout the project. We are grateful for the opportunity provided by AAFC London Research and Development Centre and Fanshawe College CARIB lab, London, Ontario, to conduct collaborative projects.
This study was funded and conducted by Solar Grants Biotechnology Inc., of which Dr. Igor Kolotilin is the founder and Chief Scientific Officer.
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Kolotilin, I. Plant-produced recombinant cytokines IL-37b and IL-38 modulate inflammatory response from stimulated human PBMCs. Sci Rep 12, 19450 (2022). https://doi.org/10.1038/s41598-022-23828-z
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Timberlake's '20/20' album sells 968K in 1st week
NEW YORK (AP) -- Justin Timberlake's comeback album has sold nearly 1 million units its first week out.
Nielsen SoundScan announced Tuesday that the singer's third album, "The 20/20 Experience," has moved 968,000 units. It's the 19th album in Nielsen's 22-year history that has sold more than 900,000 albums in its debut week.
"20/20" is Timberlake's third album and the follow-up to his multiplatinum, Grammy-winning 2006 album, "FutureSex/LoveSounds." The new CD features the pop hit "Suit & Tie."
Timberlake was strategic about promoting "20/20." He performed at the Grammy Awards, hosted and hit the stage at "Saturday Night Live," and spent an entire week on "Late Night with Jimmy Fallon." The singer also partnered with Target for the album's deluxe edition and "20/20" streamed on iTunes a week before it was released

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Closure planned in Athens Twp. for bridge replacement
ATHENS TOWNSHIP — A portion of V Drive South in Athens Township will be closed for more than three months to replace a bridge, the Calhoun County Road Department said Thursday.
The closure will begin on June 13 and will take place at the road's bridge. Work is expected to be completed Sept. 30, according to a news release.
Drivers can take S Drive South, to R Drive South, to South 47th Street and to South 46th Street to get around the construction.
A map of the detour can be seen here:
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Nickelback is a Canadian rock band formed in 1995 in Hanna, Alberta, Canada. The band is composed of guitarist and lead vocalist Chad Kroeger, guitarist, keyboardist and backing vocalist Ryan Peake, bassist Mike Kroeger, and drummer Daniel Adair.
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The winners of the "Our Scottish Borders" photography competition have been announced, and entries will be showcased in an exhibition at the Heart of Hawick from 18-21 November 2010.
Article Date: 18 Nov 2010
The Scottish Borders is taking part in the worldwide event 16 Days of Action to eliminate all forms of violence against women. The exciting and varied free event programme runs from 25 November - 10 December incorporating International Day for the Elimination of Violence against Women (25 November) and International Human Rights Day (10 December), thus making the link between women's human rights and their experience of violence.
Article Date: 11 Nov 2010
The creative heart of the Border landscape, its woods, wild moorlands, meandering rivers, historic towns and castle dotted coastline, lies in the studios and workshops of local artists and craftspeople: the painters, potters, wood turners, textile and glass artists.
Article Date: 01 Nov 2010