Artificial intelligence is dangerous. And not. My first experience of the idea that super intelligent computers could be dangerous was in the 1968 film "2001: A space odyssey" by Stanley Kubrick. On a mission to Jupiter the spacecraft's super computer, "Hal", gets increasingly emotional and erratic. Hal sounds so personal and intelligent that I will refer to this computer as "him" rather than "it". Hal decides that his own survival is more important for the mission than the survival of the humans on board. Hal kills all crew members except one, whom after some struggle succeeds in disconnecting his AI-circuits. When Hal realizes that he is going to be shut down he becomes really scared and starts singing the song "Daisy Bell" (known as "Isabella" in Swedish). This song must be the equivalent of a comforting nursery rhyme for a super computer since it was the first song ever to be sung by a computer, in 1961.
The next scary take-over attempt by AI computers that I encountered occurred 1984 in the Terminator film starring Arnold Schwarzenegger. This film has later developed into a series of films, essentially with the same plot. A computer network called Skynet that was created by the American military as an AI defence network becomes "too" intelligent and decides to launch a massive nuclear attack against humanity. Most humans are killed in the initial attack, but still many humans survive. Over the years the survivors become increasingly more successful in their war against Skynet and in 2029 Skynet is close to losing the war. At this time time-travel has become possible and Skynet sends advanced cyborg assassins (more advanced as the series of films progresses) back to 1984, before the war started. Their mission is to kill a woman, Sarah Connor. The human leader in the war in 1929 is the charismatic John Connor. Skynet's logic (which may not be that logic) is that if John Connor's mother is killed before he is born he would not exist and the humans would not have a strong leader. However, also the 2029 humans can send cyborgs back through time, these, of course, with the mission to protect Sarah Connor. Needless to say, she survives the assassination attempts.
A more recent war between humans and machines controlled by AI-computers occurs in the Matrix films, of which the first one came 1999. The year is 2199 and the war between humans and machines has almost been won by the machines. Only one free human city remains, well hidden underground. Most humans live enslaved virtual reality lives in a computer simulation called The Matrix controlled by the computers. The "reality" that these people experience is precisely the one we experience right now. In reality (well film-reality), however, these humans are kept in fluid filled incubators for obscure computer purposes. The earth has become a dystopic dark place, almost destroyed in the war between humans and machines. The free humans that still remain (called the rebels) can enter The Matrix by connecting their brains to it and in this way they can interact with humans caught in the simulated reality. When the hero (Keanu Reeves) breaks loose from his incubator he becomes "Neo" a Messiah like figure for the rebels. He eventually becomes a master of The Matrix outperforming even the fastest computer programs that are created to kill rebels that enter The Matrix.
These movies have a funny twist at the end; mankind is eventually saved from the AI take-over by a computer virus. The most potent of the programs that are created to keep the rebels out of The Matrix, a secret service type of agent in black suit called Mr Smith, becomes a virus that strives to take control over the machines. In the very moment when the machine armies find the rebel city and attacks the free humans, Neo strikes a deal with the machines. If he can defeat the computer virus, humans and machines will live in future peace. I will not tell you who won, Mr Smith or Neo.
The Swedish crime story "The Girls in the spider's web" (Swedish "Det som inte dödar oss") is a late addition (2015) to this theme. This is the fourth book in the Millennium story about the odd duo Lisbeth Salander and Mikael Blomquist. A genius computer scientist, Frans Balder, leaves his prestigious job in Silicon Valley to take care of his autistic son in Sweden. He has developed an algorithm that will lead us rapidly into the potentially dangerous point in time called the technological singularity. This occurs when AI-competent computers become so intelligent that they are smarter than humans. This is supposed to be caused by a runaway process when computers develop new and smarter computers in an accelerating process. The implicit assumption behind movies such as Terminator or The Matrix trilogy is, of course, that computers are near, or have reached this point. In the book Frans Balder is killed by an assassin who is after this super algorithm. The algorithm, however, is lost forever.
Can we learn anything from such movies and books? The most obvious wisdom to me is that we humans are fascinated by this idea. When I struggle with my own genetic algorithms and artificial neural networks a technological singularity seems very remote and maybe not even possible. But who knows? The idea of this type of accelerating development stems from Darwinian evolution thinking. Life on earth has evolved from very simple ancestral forms into ever more complex and intelligent life-forms. Modern humans are a recent life form that has taken control over earth and by far outsmarts all other species. If we consider that earth is almost five billion years old, there has clearly been an acceleration evolution of intelligence the last 50 thousand years or so. Could a corresponding accelerating evolution of artificial intelligence also occur?
The idea that evolution can occur in a run-away process comes from the great sir Ronald Fisher himself. In the early 1900-s Darwin's ideas of evolution by natural selection was becoming increasingly discredited among contemporary scientists. Darwin did not know about genes and hence he could not explain how characters could remain over generations. If characters are intermediate between two parents (like mixing two jars of paint) the individuals in a species would become increasingly more similar over generations and all variation would disappear. Such blended inheritance would make evolution as we know it impossible. The concept of genes is very different from this blended inheritance as genes are inherited intact across generations. Ronald Fisher was the most famous of the Neo-Darwinists, the scientists that unified Darwinian evolution with Mendelian genetics. In his 1930-book "The genetical theory of natural selection" he explained evolution essentially as we understand it today.
When we think about natural selection it is hard to understand how a trait like the peacock's trail possibly could evolve. A male peacock with a long tail will clearly be more at risk if a leopard attacks than a peacock with a short tail. Fisher explained the peacock's tail (and other extravagant sexual characters) with something he called runaway selection. He explained this in the following way: if a group of peahens for some reason prefer males with longer tails, long-tailed males will have a benefit since they will get more offspring than males with normal tails. If another gene makes females more likely to choose long-tailed mates, these female will also be at an advantage since they would get long-tailed (and successful) sons. In this way a genetically based tendency to choose in females would act in the same direction as the genetical base for the chosen trait in males. This system would be self-reinforcing and create run-away selection for long tails. The males with the longest tails that peacocks possible could possess would then be the ones that were most attractive to females.
Is it possible to apply this thinking to artificial intelligence? To me Fisher's ideas suggest that mere development of the capacity of computers is not sufficient for us to lose control over their abilities. We would need some kind of competition in combination with evolution, for example competition between different types of AI systems. If we turn away from fantasy and enter reality the evolution of AI systems seem far less dangerous. On the contrary, it seems like the development of computers and AI has the potential to produce the most significant contemporary improvement in our everyday lives. Clearly the benefits of this will be harvested by people and companies that work hard in early stages of this development. To me, this is precisely what Lytics.ai does.
Anders Brodin serves Lytics in a scientific advisory role within applied mathematics and machine learning. Anders is Professor in Theoretical Ecology at the Department of Biology at Lund University. He holds a Ph.D. at Stockholm University and a post doc at the Department of Applied Mathematics at University of Toronto.


NexusFile 5.3.3 Portable » SoftLab-Portable.com - Portable Soft!
NexusFile is a file manager for Windows. You can manage many files very fast and easily. NexusFile will increase your computing performance. If you still use Windows Explorer, you must try this one. Basic Functions: Copy/Move To, Copy/Cut/Past, Delete/Shred, Rename, Change Attributes/Time. Advanced Rename: Rename multiple files at once with many powerful options. Navigate Folders: Favorite Folders, Working Folders, Tabs, Smart Address Bar. Archives: manages ZIP, ARJ, RAR, ACE archive files. Dual Panels: Open two folders at once. FTP: basic FTP functions.
File Join/Split: Split large file into many small files or Join small files into a large file. Merge several text files into a file.
Shortcuts: Most functions has a Keyboard shortcut so you can do everything fast.
Multi-language: Multi-language supports with language files.
Skin: you can change the look of a program.
And many other useful functions with easy interface.


text:
Yesa robe (), also known simply referred as Yesa (), Yisan (), or Yisa (), is an ancient type of Chinese clothing worn during the Ming dynasty. It originated in the Ming dynasty but was influenced from clothing of the Yuan dynasty. It is sometimes described as being a sinicized version of the Mongol's jisün and could only be found in China. Yesa was a regular clothing in the Ming dynasty; it was initially worn in the palace and by the wealthy, and it later spread to the commoners.

Construction and design
The yesa robe is a cross-collared, long sleeved robe with narrow pleats on the lower hem; while the back of the lower part is flat, the front had two densely pleated sides with a flat middle, similar to the mamianqun. It has two hems at each sides. Compared to the Mongol Yuan's terlig, the sleeves and the bodice were wider; the pleats were also wide compared to very finely gathered skirt of the Mongol's terlig; the yesa also had no waistband; the skirt was also longer.

The absence of the threaded/braided waistband, the absence of pleats at the centre back and front of the skirt; the presence of large side pleats were the main features of the yesa; these features were developed in the Ming dynasty and were not of Mongol origins. These features also differentiated it from its precursor.

The yesa also showed sedentary life-style and Chinese cultural characteristics: its increase in length made it lose its functionality for horse-riding purposes, the flat centre back appeared as well-arranged pleats were to hard to maintain when sitting. Moreover, the folds which were originally found at the centre front of the terlig also had to disappear as it interrupted the dignity of the vertical line, a feature which was important in Chinese clothing culture.

History

Origins

During the Ming dynasty, some clothing incorporated the clothing elements from the Han Chinese and the Mongol clothing tradition; one of those mixed-elements is the yesa. The clothing known as yesa originated in the Ming dynasty, but some of its elements were either adopted from the Yuan dynasty Mongol's terlig, or directly from the Mongol's jisün clothing which is also a form of terlig. The yesa is itself a new evolution of terlig.

Ming dynasty
The yesa robe is a form of kuzhe (; clothing with trousers for riding or military style clothing); however, the design of the yesa made it no more functional for horse riding.

The yesa was worn as an informal attire by emperors, princes, ministers, and officials in their spare time during the early period of the Ming dynasty; it was worn as a formal uniforms in some occasions during the middle period of the Ming dynasty; it was worn as a casual dress worn by scholar-officials during the mid-to-late period of the Ming dynasty; and eventually it was worn by servants and commoners in the late Ming.

The Ming court eunuchs wore yesa robe in different styles, such as the red-coloured yesa robes with a qilin patch, round-collared yesa robes, and light green yesa robe, etc.

Gallery

Similar items

Jisün
Terlig
Feiyufu

See also

 Hanfu
 List of Hanfu
 Fashion in Yuan dynasty

References

Chinese traditional clothing

redpajama_set_name: RedPajamaWikipedia


text:
Karola Ruth Siegel later Ruth Westheimer maar beter gekend als Dr. Ruth (Wiesenfeld, 4 juni 1928) is een Amerikaanse televisiepersoonlijkheid. Ze werd bekend door tv-shows als Good sex! with Dr? Ruth,  Ask Dr.Ruth en What's up, Dr.Ruth.

Biografie
Karola Ruth Siegel groeide op in Frankfurt als enig kind van Irma Hanauer en Julius Siegel. Ruth Siegel werd in 1938 met een kindertransport naar Zwitserland gestuurd naar een weeshuis om te ontsnappen aan de Jodenvervolging.
Na de oorlog verhuisde ze naar het Mandaatgebied Palestina en sloot aan bij Hagana, een zionistische Joodse paramilitaire organisatie. Ze werd er sluipschutter. In 1948 werd ze gewond door een granaat tijdens de Arabisch-Israëlische Oorlog van 1948. Siegel verhuisde in 1950 naar Parijs en studeerde er Psychologie. In 1956 verhuisde ze naar de Washington Heights (Manhattan) en behaalde ze een M.A. in sociologie in 1959 aan The New School en een Ed.D in 1970 aan het Teachers College (Columbia-universiteit).

In 1965 werd ze genaturaliseerd tot Amerikaans staatsburger. Toch verkreeg ze opnieuw het Duits staatsburgerschap in 2007.

Siegel is drie keer gehuwd. Waarvan Manfred Westheimer haar laatste man was tot aan zijn overlijden in 1997.

Loopbaan
Na enige tijd voor een Planned Parenthood had gewerkt deed ze een postdoc bij Helen Singer Kaplan in het New York-Presbyterian Hospital. Ze werkte er vervolgens 5 jaar als docent en gaf ook les aan [5] Ze bleef daar vijf jaar werken als adjunct-universitair hoofddocent. Ze gaf ook les aan de Columbia-universiteit, het Lehman College, het Brooklyn College, de Adelphi Universiteit en West Point.

Daarnaast begon ze een mediacarrière met Sexually Speaking, een radioprogramma dat begon in 1980 op WYNY-FM in New York. Dit was een programma van 15 minuten om ongewenste zwangerschappen aan te pakken. De show werd populair en werd vanaf 1984 landelijk uitgezonden door NBC Radio onder de naam Dr. Ruth Show. Dit programma bleef lopen tot 1990. Daarnaast was ze vanaf 1984 ook op televisie op Lifetime-netwerk te zien onder andere in Good Sex! Met Dr. Ruth Westheimer dat in 1985 wijzigde in The Dr. Ruth Show. Vanaf 1987 liep de Ask Dr. Ruth die te zien was op meerdere zenders.
Later programma waren de The All New Dr. Ruth Show (Lifetime) vanaf 1988, What's Up, Dr. Ruth? vanaf 1989 en You're on the Air met Dr. Ruth vanaf 1990.

Door haar bekendheid was Dr. Ruth ook te gast in andere tv-programma's waaronder The Tonight Show (Johnny Carson) en Late Night with David Letterman, Between the Lions. Ze speelde ook mee in enkele films en series waaronder Electric Dreams (1984), Une femme ou deux (1985) en Dr. Ruth's House (1990).

Publicaties
Siegel schreef 45 boeken waarvan hieronder een selectie.
 Dr.Ruth's Guide to Good Sex (1983)
 Sex and Morality: Who Is Teaching Our Sex Standards (1988) met Louis Lieberman
 Dr. Ruth's Guide to Erotic and Sensuous Pleasures (1992) met Louis Lieberman
 Dr Ruth"s Guide to Safer Sex (1992)
 Dr. Ruth's Encyclopedia of Sex (1994)
 Sex For Dummies (1995)
 Power: The Ultimate Aphrodisiac (2001) met Steven Kaplan
 Dr. Ruth's Sex After 50: Revving Up the Romance, Passion & Excitement! (2005)
 Roller Coaster Grandma: The Amazing Story of Dr. Ruth. (2018)

Erkentelijkheden
 2002 - Leo Baeck-medaille
 2004 - eredoctoraat Trinity College
 2008 - ere-diploma van de Bronx High School of Science
 2019 - Documentaire Ask Dr. Ruth

Amerikaans schrijver
Amerikaans feminist
Amerikaans televisiepersoonlijkheid
Duits Holocaustoverlevende
Duits schrijver


The year 1990 was when Chile transitioned to democracy, but all of that seems a world away for 16-year-old Sofia, who lives far off the grid in a mountain enclave of artists and bohemians. Too Late to Die Young takes place during the hot, languorous days between Christmas and New Year's Day, when the troubling realities of the adult world—and the elemental forces of nature—begin to intrude on her teenage idyll. Shot in dreamily diaphanous, sun-splashed images and set to period-perfect pop, the second feature from one of Latin American cinema's most artful and distinctive voices is at once nostalgic and piercing, a portrait of a young woman—and a country—on the cusp of exhilarating and terrifying change. An NYFF56 selection. A KimStim release.


text:
/ Council and democracy
/ Lancashire Combined Authority
Lancashire Combined Authority
Over the last few months each of the councils in Lancashire has been discussing the idea of creating a Combined Authority for the county.
Lancaster City Council has backed the idea, and a formal bid has now been submitted for the Government to consider.
The shadow Combined Authority is meeting on a monthly basis with a view to becoming formally established from April 2017. Leaders have already agreed a Scheme of Governance which sets out voting arrangements and membership and the shadow Combined Authority are operating within these same principles.
What is a Combined Authority?
Lancashire's councils already work closely together but a combined authority is a formal, legal step that means councils can act as one authority in the best interests of Lancashire around the areas of economic regeneration, housing and transport.
A combined authority is:
A formal, legal step to allow councils to work more closely together in a more structured way
A move to improve economic development, skills and transport to benefit the whole county.
A combined authority is not:
A merger or take-over of councils or a unitary council
A giving-up of powers by individual councils.
The creation of a Combined Authority is something that is being replicated across the country with many bids already approved by the Government.
What are the benefits of becoming a combined authority?
A combined authority means that councils can work as one democratically accountable body around the key areas of economic development, regeneration and transport to:
Avoid duplication and provide more efficient services
Create a stronger basis for bidding for Government and EU money
Make best use of council and Government money
Speed up decision-making.
A combined authority could streamline services, be more transparent and efficient, and make it easier for Lancashire to attract investment and jobs.
We're sure you'll have plenty more questions so you can see a full list of frequently asked questions (PDF, 139KB).
Council report on the Combined Authority (PDF, 1.4MB)
Scheme of Governance (PDF, 97KB)
Governance review (PDF, 994KB)

redpajama_set_name: RedPajamaCommonCrawl


Antia, SD. & Kreimeyer, KH. (2001). The Role of Interpreters in Inclusive Classrooms. American Annals of the Deaf, 146(4), 200, 355-365. A qualitative 3-year case study followed three deaf interpreters in an inclusive school. Results of interviews indicated that, in addition to sign interpreting, the interpreters clarified teacher directions, facilitated peer interaction, tutored the deaf children, and kept teachers and special educators informed of the deaf children's progress. Differences in perception of the interpreter's role were also found.
Determining a Student's Readiness to Successfully Use Interpreting Services: This article highlights the importance of determining student readiness before placing students in interpreter-mediated educational settings. An overview of procedures developed by the Outreach Department of the New Mexico School for the Deaf are shared, including a student competency check-list , considerations for the educational team, a descriptive continuum of student readiness, and a list of educational considerations for students who are not ready for interpreter-mediated education.
Educational Interpreter Resources Toolkit: The Educational Interpreter Resources Toolkit, prepared by the 2007 - 2009 Educational Interpreting Committee (EIC), serves a resource tool to assist educational interpreters in getting valuable information for their work in the educational setting, K-12. A brief overview is given and interpreters are directed to resources related to: Laws & Policy Supporting Deaf Education, Database and Tools for Finding Additional Resources, Organizations, Books, Journals, and Articles of Interest, State-specific Educational Interpreting Guidelines, Resources to Share with School Personnel, Resources to Share with Students and Parents, and RID Resources.
Interpreting in Educational Settings (K-12): Interpreting in Educational Settings (K-12) provides a description of the role of the educational interpreter. Responsibilities that fall within the educational interpreter's domain are described, and a distinction is made between the role of an educational interpreter and other professionals (teachers, administrators, and support staff). Qualifications, compensation, and supervision of educational interpreters are also presented.
Outcomes for Graduates of Baccalaureate Interpreter Preparation Programs Specializing in Interpreting in K-12 Grade Settings: This document outlines measureable goals in different areas that baccalaureate interpreting programs offering a degree in Educational Interpreting should be addressing. In addition, it discusses the educational interpreters many roles in a school setting along with readiness markers that should be evident in the school, teacher, teacher of the Deaf and student with a hearing loss prior to hiring an interpreter.
PEPNet Tipsheet: PEPNet's Interpreting Tipsheet provides readers with information regarding the common types of services rendered by educational interpreters. Confidentiality, professional skills, and communication sight lines are all described. Attention is given to processing time, testing procedures, and the visual demands incurred during the interpreting process.
Yarger, CC. (2001). Educational Interpreting: Understanding the Rural Experience. American Annals of the Deaf, 146(1), 16-30. The present study examined the experiences, preparation, and perceptions of 63 educational interpreters employed in two rural states, using surveys and subsequent in-depth interviews with selected subjects. Only 10 of the 63 interpreters had completed interpreter preparation programs, with 5 of these having no course work related to education. None of the interpreters working in elementary or secondary schools held certification from the Registry of Interpreters for the Deaf or any other certifying body. Of the 63 interpreters, 43 were assessed using the Educational Interpreter Performance Assessment (EIPA), which uses a scale of 0-5. Test takers who score 3.5 or better are considered "coherent." The mean score on the EIPA for the 43 educational interpreters was 2.6. Respondents reported concerns about their limited understanding of American Sign Language (ASL), their ability to interpret from ASL to English, and their salaries, training, and professional status.


You come to the SAP initial screen.
Note that after each step, you can use the Back Button to quickly return to the IMG menu screen.
A Human Resource Management System enables you to set up organizational hierarchies and employee relationships and then store and administer employee data in the system. The structures of an enterprise are subdivided into organizational structures, based on an organizational plan, and administrative structures, based on the enterprise and personnel structures.
A Company is generally used in the legal consolidation module to roll up financial statements of several company codes.
Give the Company Name and Address.
Note: We need to create this request once when you save first time in user ID.
A company code is representative of a stand-alone legal entity that requires its own set of accounting records for reporting purposes.
Menu Path: SAP Reference IMG -> Enterprise Structure -> Definition->Financial Accounting -> Define, copy, delete, check Company Code.
Give Company Code identifier and additional data.
Press save button. Give all the additional detail for the Company Code.
Double click on Personnel Areas.
Give four digit identifier for your Personnel Area and additional information.
Give all the details about Personnel Area.


text:
Tripping Vittles
Authentic Vittles
Borrowed Vittles
Grandma's Vittles
A Journey Thru Life with Food, Drink, Music and Adventure
We are Westerville(STRONG)
I live and work in a town of nearly 40,000 it's not super small and not super big. We boarder Columbus Ohio but it feels further away. We have lakes and bike Paths. Schools that rank some the best in the nation. We are Westerville(STRONG) In reality we are a bit spoiled. We are not riddled with crime, our worst problem up until a few weeks ago is around the holidays we had a bunch of car break ins. Many were not even really break ins as we feel safe enough most days not to lock our cars. Our police department put up signs reminding us that we did not live in Maybury and we should always lock our car doors.
We Are Westerville
The dark world visited Westerville 4 weeks ago. We discovered a corporation was opening a third Asian massage parlor in our town. You see we were so naïve that most of us did not realize not all Asian massage establishments were created equal. That some have a really nasty seedy underbelly and when it is exposed and you start to learn about it it can be gut wrenching. The knowledge will scare you, make you shiver and then make you want to act.
Westerville acted. We mobilized ,we educated ourselves quickly on what these places are and how awful they can be. We wanted them out of our neighborhood but many of us also wanted to help the women and children affected by this thing called Human Trafficking. Many Westerville residents spoke at a community forum and then were thrilled to see our police department swoop in along with many other organizations, FBI, DHS to name a few to arrest the "bad Guys" and try and liberate some of those being held against their will.
We went to city council and where for me personally I met an incredible man. His name was Joe Morbitzer he is our Chief of Police. He took the time that night to explain all the ins and outs to the best of his ability without compromising an active investigation. Chief Morbitzer filled us in on what Westerville was doing in the legal capacity against these type of places.
The Chief than did something I was not expecting he spoke honestly from his heart about how he was effected by a Human Trafficking Survivors story. About how he felt our town could do more than just drive these businesses away but how we could help save women. Not once did he use the word try he made a statement that we as Westerville will help women who have been forced into this life of sexual servitude. He said it was Important to him to do more. I shook his hand that night and said Thank you for caring and I meant it. He was emotional and he meant what he said.
I took a few days to get back to my life as I had put most things on hold for 2 weeks while I made myself into a community activist. I was at the vet with my dog Doyle on Saturday getting some bad news. News that at the time sucked sucked worse than anything. My dog was dying and it appeared it was happening quickly.
At the same time two families in Westerville were getting news that someone they loved dearly had died that afternoon. I had not even dried my tears about my baby Doyle when my mind started racing. Please don't let it be my friends husbands. What happened? How did this happen in Westerville. Was it someone I knew or were they one of the many many police officers that came to the restaurant I worked at. How did this happen in Westerville.
How did this happen in Westerville??? Sadly it happened the way many police shootings happen. A woman in a bad domestic situation called for help the police came and everything went sideways… That is such a ridiculously simple explanation for such a tragic set of events. I know that most police view domestic disturbances as one of the most dangerous calls they ever have to go on. I know this because for a few years I lived with a police officer and we talked about her fears and that was at the top of the list.
Westerville the town that made local news a few weeks ago by organizing against human traffickers was now International News for the death of two police officers. We were a city in shock!! We did not know what to do. So a community organized again. this time to show the families of these officers we cared but also to grieve along with them.
We may seem like a big town with 40,000 people but we are a small town at heart. I have not met anyone that has not been touched in some way by this tragedy. There are those people I know who first responded to the call for help.
Many kids and young adults that knew the officers thru their work at the public schools and the library. There is the vast network of friends and acquaintances who knew and loved those men and their wives . Many businesses all over town who over the years have served those officers and their brothers and sisters in blue a pizza, a burger or a beer.
We were connected by grief. We tried to ease our pain by buying up every blue ribbon and every light bulb in every store in town and the surrounding area. People gathered in groups and wrapped trees and fences. We spoke to our neighbors maybe for the first time and with hope not the last. Drivers let other drivers into traffic, we held doors. We bought flowers and food and drink and delivered those items to the police station. As a community we grew a memorial. It started as a flower and a candle in a weeks time it grew to a shrine.
We were sad. We openly cried. Children held their parents hands and walked by police officers and said thank you.
We were a community that came together over tragedy. It felt for me like a continuation of the community pulling together just a week before. Our police chief was on TV he spoke about his friends and fellow officers Morelli and Joering. He wept openly he was filled with emotion and he reinforced something I was sure of on the day he shook my hand only a week earlier and that he is one of the good ones. His heart is huge and he cares. We needed him and he needed us.
The week felt like 10. The town came out by the 1000's as the Officers were brought to the funeral homes. We drove by those funeral homes all week seeing a lone police car stationed at the entrance keeping watch over their brothers. The job of making and delivering food to fire houses and police stations commenced. We kept serving the public and the public kept buying…Flowers, ribbon light bulbs, pizza, donuts, pasta, snacks, drinks. We as Americans always feed people when they are grieving. It is what we do because it makes us feel better and we hope a hot meal makes those hurting feel better. Food made with love and shared with love can touch a persons soul. We are Westerville(Strong)
As I waited tables this week I saw people cry. Children asked parents hard questions ? Parents tried to explain how two police officers gave the ultimate sacrifice. I saw people trying to deal with their place in this tragedy. The week passed the memorials got bigger, the town got sadder. Westerville moved closer to saying our final goodbyes.
We woke Friday morning to a crazy swing in weather the 60 degree weather and rain was gone from Thursday and the sky had turned grey and brought with it a frigid cold. I thought to myself I wonder if people will still line the streets for the procession. It was so cold and the week so long.
I sat down to have my coffee thinking about those in Westerville and what our little community had done and what today would bring. It was nice what the community did, if it had all ended there the families of our towns fallen officers knew our community cared. I made the decision no matter the weather I would stand on the route and honor these men husbands, fathers community members and protectors of their fellow citizens.
It turns out that was not how my day would go. I got a text asking me to come in early as we were going to be feeding the firemen and women who came to cover Westerville fire houses so our fire department could goto the funeral. I did not know when the day started I would come to view the simple act of delivering an honor. The fire houses were quiet when we arrived with our food. The men and women were over the top grateful and we held back tears as we gave them their food. I think we all felt it there was something that none of has experienced before about to happen. As we drove back to work it became clear that what was happening was going to be something extraordinary.The police vehicles lined the roads as far as you could see. There was a tranquility as you drove thru town that I cannot explain but I could feel it.
We went back to the pizza shop and watched the service on TV as we prepared food for our regular cast of Friday night customers. As the service ended and we could see on the TV the procession starting our phone stopped ringing.
The last turn of the procession took place in front of our pizza shop. My boss and I went out onto the corner. Cars stopped in either direction a line of police motorcycles came by lights flashing but silent. The whole world became silent as car after car came by… Minneapolis Minnesota My home state was there. Illinois, Louisiana, Indiana Kentucky and on and on. Police cars filed past us for nearly 2 hours.
The past week we were one community but Friday The whole country came to us. They came for the Morelli and Joering families they came for the police and fire departments and they came for us, the little town of Westerville.
Honestly we needed it. The count I have heard was about 2200 Vehicles and 2700 Police and fire personal. plus 1000's and 1000's of community member who lined the route for the officers last ride. A ride which stretched nearly 6 miles in length. When it started we did not know how we needed it. By the time it was over it was clearly the exact amount of love a community who lost two police officers needed.
I went back to slinging pizzas last night after an unbelievably emotional week. When I got home looking exhausted and tired. I ate some of the cookies my cousin sent me they were made with love and they warmed my heart and my belly but they did one last thing ..They reminded me there are far more good people in this world than bad.
Pass the Vittles
Hi! Welcome to Tripping Vittles
Lauri: trippingvittles
I have a love of food, travel and music and I want to share it with the world. Life is a journey of discoveries and adventure. We are all connected and my dream is to share those connections and along the path discover what makes us more alike than different.
To keep up with the Vittles follow us on all our social media sites
Creatively Authentic Vittles
Distinctly someone else's Vittles
Grandma's Recipe Box
Leia's Adventures #3 Chinese Food(Dim Sum)
Leia's Adventures #2 Brazillian Food
Buying A Tent to Escape Life
Shari Heinrich says:
A beautifully written tribute. Thank you for all you have done for our police force, and in the memories of two officers, their family, their friends. Yes, food from the heart warms the soul.
trippingvittles says:
Shari, Thank You so much for the kind words. I am a firm believer that food, music and love can heal the soul.
Karen Kaiser Mitchell says:
I have cried off and on all week and then cried again while I read your post. You have placed into words what many could not find the words to express. Thank you for taking the time to do so.
Karen Thank you for the kind words. I too have shed tears.
Follow These Vittles On
© 2020 Tripping Vittles. All Rights Reserved. Designed & built by Olive Street Design.


text:
7.62X39 Soviet By Winchester 123 Grain Super-X Soft Point Per 20 Ammunition Md: X7.6239
7.62X39 Soviet By Winchester, 123Gr, Super-X Soft Point, (Per 20)The Winchester Line Of Super-X Centerfire Rifle Ammunition continues To Be The Best You Can Buy, And It Is Still Made In The USA. The Soft Point Bullets Are Designed For Rapid, Controlled Expansion And Maximum Impact. Symbol: X76239 Caliber: 7.62X39mm Russian Bullet Weight: 123 Grains Bullet Type: Soft Point Game Selector Guide: Deer, Varmint CPX Guide Number: 2 (Light, Thin-Skinned Game) Test Barrel Length: 20" Velocity (Feet Per Second): - Muzzle: 2365 - 100yds: 2033 - 200yds: 1731 - 300yds: 1465 - 400yds: 1248 - 500yds: 1093 Energy (Foot Pounds): - Muzzle: 1527 - 100yds: 1129 - 200yds: 818 - 300yds: 586 - 400yds: 425 - 500yds: 327 Trajectory, Short Range: - 50yds: 0.5 - 100yds: 0 - 150yds: -2.6 - 200yds: -7.6 - 250yds: -15.4 - 300yds: -26.7 Trajectory, Long Range: - 100yds: 3.8 - 150yds: 3.1 - 200yds: 0 - 250yds: -6.0 - 300yds: -15.4 - 400yds: -46.3 - 500yds: -98.4Mfg No: X76239Manufacturer: WinchesterWarranty: Warranted Against manufacturers defects.Shipping InformationWeight: 0.98 pounds. Dimensions: 2.469 (L) X 5.844 (W) X 1.406 (H) inches.:SKU: 10060Total Rounds 20Price Break Discount On 10 Boxes Or More. %5 Discount Will Be applied at Check Out.
7.62X39 Soviet By Winchester, 123Gr, Super-X Soft Point, (Per 20)
The Winchester Line Of Super-X Centerfire Rifle Ammunition continues To Be The Best You Can Buy, And It Is Still Made In The USA. The Soft Point Bullets Are Designed For Rapid, Controlled Expansion And Maximum Impact.
Caliber: 7.62X39mm Russian
Bullet Type: Soft Point
Game Selector Guide: Deer, Varmint
CPX Guide Number: 2 (Light, Thin-Skinned Game)
Weight: 0.98 pounds.
Caliber 7.62X39mm
The 7.62X39mm (7.62mm Russian) was adopted by the Soviet Union in 1943, first in the SKS semiautomatic, and then in the famous AK-47 selective fire assault rifle. The concept was based on Russian experience with the 7.92X33mm Kurz in the German MKB42, the world's first assault rifle. The 7.62X39mm has more case capacity, thus higher velocity and somewhat more range than the German cartridge, but the concept remains: A short to medium-range cartridge that offers a high volume of fire as, literally, troops closed in the assault. Although officially replaced by the 5.45X39mm in the AK-74, the 7.62X39mm and its AK-47 remain in use the world over, very possibly the most prolific military rifle and cartridge ever made. The 7.62X39mm has been adapted to the AR platform, and also chambered in various civilian platforms. Firing a 123-grain bullet at 2365 fps, the 7.62X39 is low in recoil and is an excellent deer cartridge at short to medium range. — Craig Boddington


text:
Seventeen tales set on Earth featuring new adventures throughout history for the Doctor and his companions.
Styre	on May 8, 2016 at 1:32 AM
SHORT TRIPS: PAST TENSE
Considering the generally high caliber of Doctor Who historicals, the concept of this Short Trips anthology laid the paving for what could have been a brilliant collection: nothing but historicals, written by some celebrated Who authors and some relative newcomers. Unfortunately, the collection dwelled upon different things entirely…
The Immortals — Simon Guerrier — Nothing particularly interesting on display here, save the opportunity to see the Davison crew in a Hartnell-style historical. There's the expected amount of bickering and an odd ending, but really this is unmemorable. Somewhat disappointing coming from Guerrier.
Far from Home — Alison Lawson — Doesn't have a thing to do with history, but Lawson writes very well and this is her third strong effort for Big Finish. It reads like a children's story, but a good one, much like Louise Cooper's Rip Tide novella. The Doctor's fun here too. Solid, enjoyable reading.
All Done with Mirrors — Christopher Bav — The historical material is interesting and provides a unique twist on the common Marlowe-as-Shakespeare theory, but unfortunately Bav fails miserably when it comes to the regulars. The fourth Doctor is almost unrecognizable and Sarah might as well be a new companion. One of those stories that'd work better if you'd never seen Doctor Who.
CHAOS — Eric Saward — Bizarre. Saward appears to be trying his hand at a mood piece, showing the Doctor subjected to various dream sequences and tortures. Unfortunately his prose is Targetesque and doesn't support what he's trying. Still, he clearly knows the regulars and writes some solid dialogue. Not sure what this has to do with the theme, though…
Ante Bellum — Stephen Hatcher — Cliched WWI spy thriller with an obvious inversion. The regulars are incredibly generic — the only reason this appears to be Ace is her hatred of Germans, and the seventh Doctor could be any of his fellows. The plot is unimpressive as well; thus far the collection has been decidedly average.
The Thief of Sherwood — Jonathan Morris — Excellent stuff, dealing with both Doctor Who historicals and Doctor Who's own history. Not only does Morris outline a believable Hartnell historical involving Robin Hood, he perfectly captures the tone of each of the various types of review article that would follow. The Television Companion analysis referencing a fake Gary Russell Shada article from 1981 had me laughing out loud. Brilliant, though it's really little more than a fan gratification piece.
Come Friendly Bombs… — Dave Owen — It's really little more than a lecture on the firearms debate, but Owen introduces a sufficient degree of moral ambiguity to make it work. It'd have to be Pertwee telling this to Jo, wouldn't it? And the end is as good as it is necessary. Strong stuff.
Graham Dilley Saves the World — Iain McLaughlin & Claire Bartlett — As I don't know anything about cricket, I agreed wholeheartedly with Peri.. not sure if cricket fans find the other side as funny, though. Of course the regulars are their wonderful BF selves, though the ending is painfully overdone as silly drunk people annoy the hell out of me. Worth reading, though insignificant as with a lot of the material in this volume.
Bide-a-Wee — Anthony Keetch — A great followup to Keetch's hilarious "Twin Piques" in the Zodiac anthology. This is a great discussion on the difficulties found when one (literally) lives in the past, and Keetch's Hartnell is absolutely perfect. Perfect leisurely reading material and so much fun to read. Great work.
Mortlake — Mark Wright — A nice little piece about the forgotten figures of history, and the unique position the Doctor and his companions are in to do something about the situation. Padiel is hilarious, too. Wright clearly knows and loves these BF regulars.
White Man's Burden — John Binns — Binns takes a step back from the esoteric style he'd adopted in previous anthologies and scores a hit. The fifth Doctor and Turlough section is effective enough on its own, but counterpointing that philosophy against that of the Hartnell historicals with a beautifully-portrayed Ian and Barbara makes the story something better.
Of the Mermaid and Jupiter — Ian Mond & Danny Heap — For once two authors take a stab at the NA characters and atmosphere and actually get it right, but unfortunately the plot is unreasonably silly. True events these may have been, but just because you're aware of the giant logical flaw in your story doesn't make it acceptable. Still, this is a great Benny/Doctor pairing.
The Man Who Wouldn't Give Up — Nev Fountain — Ha! The sixth Doctor travels through time pretending to right wrongs just so he can get away from Mel's diet plan! Honestly, I must be the only person that doesn't think Colin Baker looked that fat on television, but this is hilarious.
One Small Step… — Nicholas Briggs — Briggs expertly captures the Troughton regulars here, but as with Mond & Heap's story mentioned before, the plot doesn't hold up. For some reason the Doctor is allowed to change history here without any consequences and without explanation. Speaking mysteriously of great lengths and broken rules doesn't justify a gigantic plot hole, especially not when the entire story revolves around it.
To Kill a Nandi Bear — Paul Williams — Inappropriate for a short story, as Williams attempts to portray the first-ever TARDIS trip (that we've seen) to sub-Saharan native Africa and cram character and setting into nine pages. Fleshed out, and with a decent ending, this would make a good novella, but as a short story it's just rushed and sloppy.
Fixing a Hole — Samantha Baker — Why? The title points the way to a needless character piece that attempts to explain "A Fix with Sontarans," as though it needed doing. Reconciling Tegan and the Doctor is a terrible idea, no matter how good the writing (and this is average) — someone should explain to Baker that her departure on a down note was actually a good idea. This also has nothing whatsoever to do with the theme of the collection.
That Time I Nearly Destroyed the World Whilst Looking for a Dress — Joseph Lidster — After the pointless hole-fixing that went on in the previous story, here Lidster goes over the top and explains a continuity hole in Jonathan Morris's fictional Thief of Sherwood story! Lidster is clearly sending up the whole threats-to-the-web-of-time concept by interfering in the stories of his fellow authors, and the results are hilarious — though unfortunately the first-person narrative starts to grate after a while. Please tell me editor Ian Farrington knew what he was doing when he put this story immediately after Fixing a Hole…
For all of this anthology's claims to depict the Doctor's "enjoyment of seeing history happen," this rarely happens in Short Trips: Past Tense. It's mostly a grouping of prevent-history-from-being-changed stories or stories which have nothing whatsoever to do with Doctor Who's historical concepts. This is not to say there isn't any strong material on display here, but on the whole this collection is little better than average and it definitely wastes the potential of its framing device. Again, I'm not sure if it's worth the price, but if you can get it cheaper it's worth checking out.


1 THB = 0.11 QAR at the rate on 2019-04-20 .
The page provides data about today's value of one baht in Qatari Rials. The interactive form of the currency calculator ensures navigation in the actual quotations of world currencies according to "Open Exchange Rates" and displays the information in a graph.
The online converter will clarify the ratio between different currencies. For example, you can instantly convert 1 THB to QAR based on the rate offered by "Open Exchange Rates" to decide whether you better proceed to exchange or postpone currency conversion until better times.
The value of 1 THB in Qatari Rials for the week (7 days) decreased by: -0.000475 QAR (zero rial zero dirham).
The value of 1 THB in Qatari Rials for the month (30 days) decreased by: -0.000343 QAR (zero rial zero dirham).
The value of 1 THB in Qatari Rials for the year (365 days) decreased by: -0.002014 QAR (zero rial zero dirham).


Published by B.P, at Tuesday, August 28th , 2018 - 17:10:49 PM, in Salle À Manger category.
Published by M.R, at Tuesday, August 28th , 2018 - 17:23:49 PM, in Salle De Bain category.
Published by M.F, at Tuesday, August 28th , 2018 - 08:41:49 AM, in Salle À Manger category.
Published by John Who, at Tuesday, August 28th , 2018 - 13:07:49 PM, in Salle À Manger category.
Published by V.B, at Tuesday, August 28th , 2018 - 15:26:49 PM, in Jardin category.
Published by M.F, at Tuesday, August 28th , 2018 - 13:05:49 PM, in Salle De Bain category.
Published by A.F, at Tuesday, August 28th , 2018 - 11:34:49 AM, in Jardin category.


text:
Q: How to reproduce tor build using gitian? Please refer me to the steps to reproduce a tor build using gitian. I have been trying to build the tor browser bundle using steps mentioned at the following link-
https://trac.torproject.org/projects/tor/wiki/doc/TorBrowser/BuildingWithGitian
But I keep getting errors, and I did not receive a reply from help@rt.torproject.org

~/tor-browser-build/gitian-builder/inputs
  ~/tor-browser-build/gitian-builder/inputs object
  ebcbfd6cdc29372909079d0345185733d47d90d4 type commit tag
  tor-browser-38.2.0esr-5.0-1-build2 tagger Mike Perry
   1439157725 -0700
5.0-build2. gpg: Signature made Sun 09 Aug 2015 06:02:10 PM EDT using RSA key ID D2F1E186 gpg: Good signature from "Mike Perry
  " gpg:                 aka "Mike Perry
  (Regular use key) " gpg:                 aka
  "Mike Perry (Regular use key) " gpg:
  aka "Mike Perry " gpg: Note: This key has
  expired! Primary key fingerprint: C963 C21D 6356 4E2B 10BB  335B 2984
  6B3C 6836 86CC
       Subkey fingerprint: CC69 3F6C D7AA 6B8E EC40  EC28 4102 F895 D2F1 E186 error: could not verify the tag
  'tor-browser-38.2.0esr-5.0-1-build2' tor-browser: verification of tag
  tor-browser-38.2.0esr-5.0-1-build2 against
  /home/tor/tor-browser-build/tor-browser-bundle/gitian/gpg/torbutton.gpg
  failed!
You should run 'make prep' to ensure your inputs are up to date make:
  *** [build] Error 1

I ran 'make prep' and then 'make' but I still get the same error.
Edit:
After manually updating the key, but error still not resolved.
tor@tor-VirtualBox:~$ gpg -k
/home/tor/.gnupg/pubring.gpg
----------------------------
pub   8192R/683686CC 2013-09-11
uid                  Mike Perry <mikeperry@endarken.info>
uid                  Mike Perry <mikeperry@unencrypted.info>
uid                  Mike Perry (Regular use key) <mikeperry@fscked.org>
uid                  Mike Perry (Regular use key) <mikeperry@torproject.org>
sub   4096R/0F129402 2015-09-07 [expires: 2016-09-11]
sub   4096R/ACC0A961 2015-09-07 [expires: 2016-09-11]

tor@tor-VirtualBox:~$ gpg --fingerprint 683686CC
pub   8192R/683686CC 2013-09-11
      Key fingerprint = C963 C21D 6356 4E2B 10BB  335B 2984 6B3C 6836 86CC
uid                  Mike Perry <mikeperry@endarken.info>
uid                  Mike Perry <mikeperry@unencrypted.info>
uid                  Mike Perry (Regular use key) <mikeperry@fscked.org>
uid                  Mike Perry (Regular use key) <mikeperry@torproject.org>
sub   4096R/0F129402 2015-09-07 [expires: 2016-09-11]
sub   4096R/ACC0A961 2015-09-07 [expires: 2016-09-11]

A: I doubt you're doing anything wrong.  It's failing because you're using an expired gpg key for Mike Perry.  It's probably that the script doesn't understand what caused the error, so it asks for make prep.
Appears Mike uses a fixed main key but gives his subkeys and expiration time of 1 year and replaces them.  Amusingly this Tor Project pages get it wrong too.
Just download his new key here and gpg --import it.  Or simply run :
gpg --keyserver pgp.mit.edu --recv 29846b3c683686cc

After that, running gpg --fingerprint 683686CC should show :
pub   8192R/683686CC 2013-09-11
      Key fingerprint = C963 C21D 6356 4E2B 10BB  335B 2984 6B3C 6836 86CC
uid                  Mike Perry <mikeperry@endarken.info>
uid                  Mike Perry <mikeperry@unencrypted.info>
uid                  Mike Perry (Regular use key) <mikeperry@fscked.org>
uid                  Mike Perry (Regular use key) <mikeperry@torproject.org>
sub   4096R/0F129402 2015-09-07 [expires: 2016-09-11]
sub   4096R/ACC0A961 2015-09-07 [expires: 2016-09-11]

You'll notice the new keys now expire in 2016.
You can trust this new key really belongs to Mike Perry because it's actually the same base key with the same fingerprint.  It's only the subkey's that changed.  In fact, there is a good chance Mike does this precisely so that (a) scripts can update his key but (b) if his key is compromised then people cannot use the compromised key for more than 1 year, and (c) he'd keep the main key air gapped probably too.

redpajama_set_name: RedPajamaStackExchange


text:
Immunization with Streptococcus suis bacterin plus recombinant Sao protein in sows conveys passive immunity to their piglets
Kai-Jen Hsueh1,2,
Li-Ting Cheng2,
Jai-Wei Lee3,
Yao-Chi Chung2,
Wen-Bin Chung1 &
Chun-Yen Chu2
Streptococcus suis (S. suis) causes arthritis, meningitis, septicemia, and sudden death in pigs and is also an zoonotic agent for humans. The present study demonstrated that immunization with recombinant Sao-L (surface antigen one-L, rSao-L) protein from a strain of S. suis serotype 2 in pigs was able to increase cross-serotype protection against S. suis serotype 1 and 2 challenge. Since weaning piglets are more susceptible to S. suis infections due to the stresses associated with weaning, prepartum immunization in sows may convey passive immunity to piglets and provide protection.
Pregnant sows were immunized with a vaccine containing inactivated S. suis serotype 2 plus rSao as the antigens. Blood samples were collected from their piglets after birth for analysis of antigen-specific antibody titers and levels of various cytokines. Results demonstrated that the titers of S. suis and rSao-specific antibodies were significantly (p < 0.05) higher in the vaccinated piglets in comparison with that of piglets in the control group. The serum levels of interferon (IFN)-γ, interleukin (IL)-4, IL-6, and IL-12 were significantly (p < 0.05) increased in piglets born from vaccinated sows when compared to piglets from unvaccinated sows. In addition, piglets were challenged by heterologous and homologous S. suis. All piglets from unvaccinated sows developed severe symptoms of bacteremia, fever, anorexia, depression, and arthritis. On the other hand, piglets from vaccinated sows had significantly (p < 0.05) reduced clinical symptoms and lesion score (by 75 and 81%).
Our results revealed that immunizing pregnant sows with the vaccine containing inactivated S. suis bacterin plus rSao as the antigens is able to enhance passive immunity against heterologous and homologous S. suis challenge in their piglets.
Streptococcus suis (S. suis) causes a spectrum of diseases, including arthritis, meningitis, septicaemia and sudden death, in pigs and is recognized as an important zoonotic agent for humans [1]. A total of 33 serotypes of S. suis, according to the structure of its capsular polysaccharides (CPS), have been identified [2]. Serotypes 1, 1/2, 2, 7, 9, 14 and 22 are more virulent than others and can be isolated worldwide, including the United States, Canada, Europe, and New Zealand [3]. In Taiwan, more than 80% of the clinical cases are S. suis positive [4], and most of them are infected with serotypes 1 and 2 [5]. The efficacy of many commercially available inactivated S. suis whole-cell vaccines left something to be desired due to the fact that the protection is only limited to homologous strains [6]. The CPS has been shown to play a pivotal role in the pathogenesis of S. suis, and that antibody against CPS (serotype 2) is essential for full protection from homologous challenge [7]. CPS is a poor immunogen due to its thymus-independent property and cannot induce sufficient immune response [8]. Study showed that coupling S. suis type 2 CPS to tetanus toxoid can induce T cell-dependent response, inducing IgM and IgGs for protection against challenges in mice and pigs [9, 10]. In addition, immunization with certain virulence factors, such as suilysin, muramidase-released protein (MRP) and extracellular factor (EF), has been demonstrated to protect pigs from challenge with homologous or heterologous strains of S. suis. However, discrepant results were reported when different virulence factors are expressed by the prevalent S. suis strains [11]. Therefore, researching for a more immunogenic antigen that is commonly expressed by most S. suis strains is imperative.
Many surface proteins are involved in the pathogenesis of Gram-positive bacteria and have been shown to elicit strong immune responses [12–14]. Immunization with recombinant S. suis SsnA protein (surface-anchored DNA-nuclease) formulated with aluminum hydroxide was able to protect mice from S. suis infection [15]. A new surface protein of S. suis, Abpb (amylase-binding protein B) is recently identified and immunization with recombinant Abpb provided effective protection against S. suis [16]. In addition, a Lam protein (CDS0330) was expressed on the cell surface of S. suis 2 and was found to bind laminin in vitro. Immunization with recombinant CDS 0330 protein resulted in 50% survival rate of mice from S. suis 2 infection [17]. S. suis, Sao (surface antigen one), a common surface protein containing an Leu-Pro-X-Thr-Gly (LPXTG) motif, mediates many virulence factors during the course of infection. Sao is highly conserved among S. suis strains and has become a potential antigen for the development of effective vaccines against S. suis [13]. Sao protein is encoded by three allelic variants of gene with different lengths, Sao-S (1.5 kb), Sao-M (1.7 kb) and Sao-L (2.0 kb), respectively, and Sao-M is the most prevalent variant [14]. Immunization with rSao protein was able to elicit strong humoral antibody responses, decrease clinical signs and bacterial dissemination, increase survival rates, and confer cross-serotype protection in mouse and pig vaccination protocols [18], indicating that rSao is a suitable antigen for subunit vaccine development.
We previously demonstrated that immunization with recombinant Sao-L protein (rSao-L) from a strain of S. suis serotype 2 in pigs was able to increase antigen-specific antibody titers, the percentages of CD8+ and CD4+/CD8+ double-positive T cells, and cross-protection against S. suis serotype 1 heterologous challenge [19]. Since weaning piglets are more susceptible to S. suis infections due to the stress associated with weaning, prepartum immunization in sows may convey passive immunity to piglets and provide protection. This approach has been proven effective in preventing common swine diseases such as atrophic rhinitis [20], food-and-mouth disease [21] and classical swine fever [22]. Moreover, immunizing pregnant sows with a vaccine containing recombinant Pasteurella multocida toxin (rsPMT) plus Pasteurella multocida type A bacterin as the antigens significantly increased neutralizing antibody titer in colostrum when compared to pregnant sows immunized with the vaccine containing rsPMT only [20]. The combination of recombinant antigens with inactivated bacteria may provide extra antigens and elicit more broaden ranged protection in immunized animals. In the present study, pregnant sows were immunized with the vaccine containing inactivated S. suis serotype 2 plus rSao-L as the antigens. Passive immunity in their piglets was analyzed by examining serum antigen-specific antibody titers, levels of various cytokines, including interferon (IFN)-γ, interleukin (IL)-4, IL-6, IL-8, IL-12, and tumor necrosis factor (TNF)-α, and clinical signs after heterologous and homologous challenges with S. suis serotype 1 and 2.
Bacterial strains and expression of rSao
S. suis serotype 1 and 2 strains were obtained from the Pingtung County Animal Disease Control Center, Pingtung, Taiwan. Bacteria were grown in brain-heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Spark, MD, USA) at 37 °C. The rSao was expressed as previously described [19]. The size of PCR product from the S. suis strain BCRC 14750 (ATCC 43765) was 2013 bp. Primers for rSao gene were designed according to GenBank accession no. JF 810176 (Sao-F: GCGGGAT CCATGAATACTAAGAAATGGAG and Sao-R: CAGAA GCTTGAACTAATTTACGTTTACGTG). The primers contained restriction enzyme (Bam HI/Hind III) cutting sites and the PCR product was cloned into the expression vector pET32a according to the manufacturer's instructions (Novagen, Darmstadt, Germany). E. coli strain BL21 (DE3) harboring the recombinant plasmid was cultured in Luria-Bertanior modified medium (tryptone: 6 g/L, soytone: 1.5 g/L, yeast extract: 7.5 g/L, KH2PO4: 1.2 g/L, K2HPO4: 0.6 g/L, glucose: 2 g/L) at 37 °C until the absorbance reached 0.6 at 600 nm. Isopropylthio-β-D-thiogalactose (IPTG) (Amresco, Ohio, USA) was added at a final concentration of 1 mM, and the culture was grown for 4 h with agitation. The resulting culture was ultrafiltrated by Vivaspin 20 100KDa MWCO (GE Healthcare, UK). The concentrated culture was analyzed by SDS-PAGE and Western blotting using the antisera from rabbits immunized with purified rSao. Contamination of endotoxin in the crude rSao was examined using Limulus Amebocyte Lysate test, LAL (CAPE COD, MA, USA) and the result was negative (no clotting, indicating that the concentration of endotoxin is less than 0.125 IU/mL).
Purification of rSao
E. coli expressing rSao were centrifuged at 4 °C, 14,000 xg for 20 min. The pellet was resuspended in denaturing lysis buffer, sonicated using Misonix Sonicator S-4000 (Misonix, New York, USA), and filtered (0.22 μm). The rSao was purified using the ProfiniaTM Protein Purification System (BIO-RAD, California, USA) according to the manufacturer's instructions. Urea was incrementally dialyzed out of the samples, from 6 M, 5 M, 4 M, 3 M, 2 M, and 1 M urea to 0.85% saline, using 12,000–14,000 MWCO Dialysis Membrane (Cellu•Sep® MFPI,Orange Scientific, Braine-l'Alleud, Belgium). Thereafter, BSA was used as the standard for determining the concentration of rSao.
Preparation of vaccines
Formalin-inactivated S. suis serotype 2 bacteria and rSao in the aqueous phase final concentration in the vaccine for S. suis and rSao was 1x109 CFU/mL and 100 μg/mL, respectively. The oil vaccine prepared with internal aqueous phase was emulsified with 25% of water-in-oil-in water (w/o/w) adjuvant containing two inactivated antigens of S. suis and rSao: oil phase: external aqueous phase ratios of 1:1:2 [19, 23]. The vaccine did not contain any viable microorganisms, which was confirmed by sterility test.
Immunization of pregnant sows
Ten healthy pregnant sows, without previous exposure to S. suis (S. suis 2 antibody negative), were divided into two groups (n = 5 each). The vaccinated group was intramuscularly immunized with S. suis serotype 2 plus rSao vaccine (2 mL) 6 and 2 (boost) weeks before parturition. The other group was injected with an equal volume of sterile saline as the unvaccinated control. Animal experiments were conducted in a private pig farm in Tainan County, Taiwan, with informed consent from the owner to conduct research. The pigs were selected and randomly assigned to the treatment groups for the vaccine trial. All experimental protocols for animal trials were approved by the Animal Care and Use Committee, National Pingtung University of Science and Technology (NPUST). The experiments were conducted based on the Ethical Rules and Law of NPUST.
Assessment of passive immunity in piglets
Blood samples (3–4 mL) were collected from the superior vena cava 2, 4, and 6 weeks after parturition from 5 healthy piglets randomly selected from each groups (n = 5) for determining the serum antibody titer and cytokine levels. Five weeks after parturition, 3 piglets were randomly selected from each group (n = 3) and challenged i.p. with 2 mL (1.06 × 109 CFU/mL) of live S. suis serotype 1 (P1) or S. suis serotype 2 (P2) bacteria. The piglets were monitored daily for clinical signs, body temperature (fever was defined as anal temperature≧40 °C), and bacteremia. Seven days after the challenge, the piglets were i.m. injected with Stresnil (0.1 mL/kg), sacrificed, and dissected. Thereafter, pathological examination was performed and lesion score was calculated based on the area of lesions in an organ, where no lesion = 0, lesion area < 33% = 1, lesion area 33–66% = 3, lesion area > 66% = 5. The necropsy samples of various organs were cultured for bacteriological analysis.
Indirect enzyme-linked immunosorbent assay (ELISA) for serum antibody titers
Flat-bottomed 96-well plates were coated with either S. suis serotype 2 bacteria (1.25 μg/mL) or purified rSao (1.25 μg/mL) in the coating buffer (15 mM Na2CO3, 35 mM NaHCO3, 3 mM NaN3, pH = 9.6) at 4 °C overnight. The serum antibody titers were determined as described [19], where S/P ratio = (Sample – NCˉ x) / (PCˉ x – NCˉ x) (NC: negative control, PC: positive control). To obtain positive serum, S. suis-negative SPF pigs were challenged with 2 ml (1 × 109 CFU/ml) S. suis 2 and serum was collected 14 days later.
Detection of serum cytokine levels
The serum cytokine levels, including IFN-γ, TNF-α, IL-4, IL-6, IL-8 and IL-12, were analyzed using the MILLPLEX® MAP Kit (EMD Millipore Corporation, Billerica, MA, USA) according to the manufacturer's instructions. The plates were read by MAGPIX® plate reader at 470–565 nm (EMD Millipore Corporation).
Detection of bacteremia by multiplex PCR
The detection of bacteremia in blood and organ samples after challenge was carried out using bacteriology and the Multiplex PCR based method. Primers for S. suis cps II and Sao gene were designed by Smith [24] and Hsueh [19], respectively: (cps II-F: GGCGGTCTAGCAGATGCTCG and cps II-R: GCGAACTGTTAGCAATGAC) (Sao-F: GCGGGATCCATGAATACTAAGAAATGGAG and Sao-R: CAGAAGCTTGAACTAATTTACGTTTACGTG). Bacterial DNA was isolated using the Blood & Tissue Genomic DNA extraction Miniprep System kit (Viogene, Sunnyvale, CA, USA). The genes were amplified by 30 PCR cycles consisting of 1 min denature at 94 °C, 2 min annealing at 56 °C and 2 min elongation at 72 °C.
All data were expressed as mean ± S.E.M. and were compared using Student's t-test. A p-value of < 0.05 was considered significant.
Incidence of fever and bacteremia after challenge
Two piglets from the control group had fever on the first (≧40 °C) and second (≧41 °C) day after challenge. One of the piglets with fever was confirmed to have bacteremia and died on the third day after challenge. In contrast, all piglets from the vaccinated group remained healthy and no fever was observed (Table 1). Results of multiplex PCR demonstrated that all the piglets in the control group had bacteremia during the first two days after challenge (Fig. 1). On the other hand, only one piglet in the vaccination group had bacteremia on the second day (Table 1) and seventh day (Table 2) after challenge. The difference between the vaccinated and the control group was marginal significant (P = 0.058 and P = 0.140) (Tables 1 and 2).
Table 1 Fever and bacteremia of piglets after challenge with heterologous Streptococcus suis serotype 1 bacteria
Detection of Sao and cps II genes in bacteremia samples by multiplex PCR. M: 100 bp DNA Ladder; Lane 1–6: n of 3 on day 1 and 2 after challenge as control group; Lane 7: Vaccinated group, two days after challenge; Lane 8: S. suis serotype 1 as the positive control
Table 2 Fever and bacteremia of pig after challenge with homologous Streptococcus suis serotype 2 bacteria
Serum antigen-specific antibody titers
Results indicated that antigen-specific antibodies, IgG against S. suis serotype 2 bacteria and rSao, in serum were significantly (P < 0.05) increased in piglets born from vaccinated sows in comparison with those in piglets born from the unvaccinated sows at 2, 4, and 6 weeks after parturition (Fig. 2a and b). The titer of serum IgG against S. suis 2, but not rSao, was remarkably reduced for piglets 6 weeks of age.
The antibody response of (a) S. suis 2 (b) rSao-specific IgG in the serum of piglets from sows with or without (n = 5 each) prepartum immunization with inactivated S. suis 2 + rSao. Data are represented as Mean ± SEM of peak absorbance values as determined by indirect ELISA performed on diluted (1:500) serum samples. S/P ratio = (sample – Nˉx) / (PCˉx – NCˉx). Differences between groups were analyzed by Student's t-test, statistically significant difference (P < 0.05) in comparison to the control group
Serum cytokine levels
The concentration of IFN-γ in serum was significantly (P < 0.05) increased in the piglets from vaccinated sows at 2 and 4 weeks of age when compared to that of piglets from unvaccinated sows (Fig. 3a). Whereas the concentrations of IL-4 (Fig. 3b), IL-6 (Fig. 3c), and IL-12 (Fig. 3d) were significantly (P < 0.05) elevated in piglets from vaccinated sows at 2, 4, and 6 weeks after parturition when compared to those of piglets from unvaccinated sows. In contrast, piglets from vaccinated sows had a significantly (P < 0.05) higher concentration of IL-8 at 2, but not 4 and 6 weeks of age (Fig. 3e), when compared to piglets from vaccinated sows. In the case of TNF-α, no differences were observed between the two groups at all the time points (Fig. 3f).
The concentration of cytokine (a) IFN-γ (b) IL-4 (c) IL-6 (d) IL-12 (e) IL-8 (f) TNF-α in piglets from sows with or without (n = 5 each) prepartum immunization with inactivated S. suis 2 + rSao. Data are represented as Mean ± SEM of the serum concentration of each cytokines determined by MILLPLEX® MAP Kit. Differences between groups were analyzed by Student's t-test, statistically significant difference (P < 0.05) in comparison to the control group
Clinical symptoms and pathological lesions after sacrifice
According to the pathological examination, piglets from sows in the control group showed various clinical signs, including fever, anorexia, and gross lesions in the peritoneal cavity, joints and lungs, in response to the challenge. In contrast, piglets from sows in the vaccinated group remained healthy and only mild lesions were observed. Results from the lesion score indicated that prepartum vaccination in sows significantly (P < 0.05) reduced the lesion score by 75% (7 v.s. 27) and 81% (4 v.s. 21) in their piglets after challenge with S. suis serotype 1 and 2 bacteria, respectively (Tables 3 and 4). Presence of S. suis bacteria in the necropsy samples of various organs (synovial fluid, peritoneal cavity and lungs) was verified and confirmed, using multiplex PCR, to be the S. suis serotype 1 and 2 strains used for challenge (Tables 1 and 2).
Table 3 Percentage of pathological lesion score after challenge with heterologous Streptococcus suis serotype 1 bacteria
Table 4 Percentage of pathological lesion score after challenge with homologous Streptococcus suis serotype 2 bacteria
Histopathological examination
All piglets survived from challenge were sacrificed and dissected on the 7th day after challenge. Thereafter, histopathological examination was performed. Piglets from unvaccinated sows displayed signs of lesions in the brain and lung, including slight vascular congestion on the brain meninges, influx of neutrophils, lymphocytes, and shedded epithelial cells in the lung alveolar space, and accumulation of neutrophils in the bronchium cavity, indicating the development of bronchopneumonia, hemorrhagic bronchial pneumonia, and lung abscess (Fig. 4a and b). In contrast, above-mentioned signs of lesions were not observed in piglets from vaccinated sows after dissection.
Results of histopathological examination. Piglets from sows with or without prepartum immunization with inactivated S. suis 2 + rSao were challenged with (a) heterologous S. sui serotype 1 and (b) homologous S. sui serotype 2 bacteria at the 5 weeks of age. The histopathological examination was performed on the 7th day after the challenge. Control pigs manifested severe brain and lung lesions, including slight vascular congestion on the brain meninges and shedded epithelial cells in the lung alveolar space
Piglets are more susceptible to infections due to the stress associated with weaning and the immature immune system during the early stage of life. Prepartum immunization in sows has been proven safe and efficiently conveys protection to piglets through the transfer of passive immunity in milk. Prepartum vaccination with a subunit vaccine containing recombinant Pasteurella multocida toxin proteins as the antigen in sows was able to enhance the level of neutralizing antibodies, reduce the degree of turbinate atrophy, and increase the survival rate in piglets with experimentally induced progressive atrophic rhinitis (PAR) [20]. Immunizing pregnant sows (52 and 32 days before parturition, respectively) with a type O1 foot and mouth disease oil emulsion vaccine significantly increased the serum level of neutralizing antibodies in piglets 3 days after birth [21]. Moreover, 3-week-old piglets born from sows immunized with a classical swine fever vaccine (E2) twice after insemination had significantly higher maternal antigen-specific antibodies as indicated by the ND50 (neutralization dilution 50%) [22].
We previously demonstrated that immunization with the rSao (from serotype 2 S. suis) protein expressed by bioreactors formulated with w/o/w adjuvant increased antigen-specific antibody titers, the percentages of CD8+ and CD4+/CD8+ double-positive T cells, and successfully provided cross-serotype protection against the challenge of heterologous S. suis serotype 1 bacteria in pigs [19]. In combination with Quil A, rSao can reduce clinical symptoms after challenge infection with heterologous S. suis and stimulates strong opsonizing antibody responses [18]. A previous study has indicated that the pregnant sows immunized with a PAR vaccine containing three short fragments of recombinant subunit Pasteurella multocida toxin (rsPMT) in combination with Pasteurella multocida type A bacterin (rsPMT + P. multocida bacterin) as the antigens can elicit higher neutralizing antibody titer in colostrum than pregnant sows immunized with the vaccine containing rsPMT only (1:101 v.s. 1:80) [20]. The immunization protected 60% of piglets born from these sows after challenged by intramuscular injection with five-fold lethal dose of authentic PMT. The results implied that combination of recombinant antigen with inactivated bacteria as the antigen may enhance the immune responses elicited by the vaccine. In addition, virulence factors of S. suis other than Sao, such as suilysin, muramidase-released protein (MRP), and extracellular factor (EF), also play a role in the pathogenesis of S. suis. Immunization with a subunit vaccine containing these virulence factors (MRP+/EF+) from S. suis serotype 2 as the antigens has been demonstrated to protect pigs from challenge of homologous or heterologous S. suis strains [25]. Addition of inactivated S. suis bacteria containing these surface virulent factors into the vaccine formula may provide extra antigens and elicit broader protection in immunized animals. Therefore, pregnant sows were immunized with the vaccine containing inactivated S. suis serotype 2 plus rSao as the antigens and the passive immunity against heterologous S. suis serotype 1 infection in their piglets was investigated in the present study.
The serum titer of IgG against rSao and S. suis serotype 2 in piglets from vaccinated sows remained significantly higher than that of piglets from unvaccinated sow until 6 weeks of age (Fig. 2a and b). After challenge with live S. suis serotype 1 bacteria, the lesion score of piglets from vaccinated sows was reduced by 75%. These results implied that immunization with rSao plus inactivated S. suis serotype 2 in pregnant sows enhanced the passive immunity against S. suis and provided cross-protection in their piglets. The Sao protein is highly conserved among S. suis strains and Sao-specific antibodies have been shown to cross-react with 28 different serotypes of S. suis [13]. The lesion score was reduced by 60% in pigs immunization with rSao alone after challenge with live heterologous S. suis bacteria [19]. Results from the present study revealed that the lesion score was reduced by 75% in piglets born from sows immunized with inactivated S. suis bacteria plus rSao after challenge with live heterologous S. suis bacteria.
In piglets produced by vaccinated sows, the antigen-specific IgG titer remained high until 6 weeks of age (Fig. 2a and b). This is in agreement with a previous study in which pregnant sows were immunized with the autogenous S. suis bacterin and MRP-specific IgG remained detectable until 6 weeks of age [25]. However, a remarkable reduction of the antigen-specific IgG titer in serum was observed in the 6 weeks of age in comparison with the titers in the 2 and 4 weeks of age, which is possibly due to the half-life of IgG absorbed from colostrum. The IgG concentrations in piglets at 24 h of age ranged from 18.7 to 39 mg/mL and gradually decreased to 6.3 mg/mL at 5 to 6 weeks of age [26]. The w/o/w adjuvant used in our vaccine has been shown to stimulate both humoral and cell-mediated arms of immune responses [17, 23].
Compared to piglets from sows in the control group, piglets from sows in the vaccinated group had significantly (P < 0.05) increased levels of IL-4, IL-6, IL-12, and IFN-γ in serum after parturition. The source of these cytokines in piglets' circulation could be mainly from the absorption of cytokines in colostrum before gut closure. IL-4 and IL-6 plays a role in enhancing Th2-type immune responses, B cell proliferation, and antibody production [27]. On the other hand, IL-12 and IFN-γ are the key cytokines that lead immune responses to Th1-type. IL-12 activates immune cells, such as NK cells and naïve CD4+ T cells, to secret IFN-γ, which in turn facilitate macrophages maturation [28–32]. Therefore, the higher concentration of cytokines in serum may contribute, at least partially, to the enhanced resistance to challenge with live S. suis bacteria in piglets from vaccinated sows. It is unclear why the level of serum IL-8 was increased only at 2 weeks after parturition in piglets from sows in the control group. IL-8 is a potential chemoattractant that is up regulated during the course of intramammary infections [33]. However, whether the sows had mastitis was not determined in our study. The serum levels of TNF-α were low and not different between the two groups, which is probably due to the short half-life of this cytokine [27].
Results of the histopathological examination revealed that piglets from unvaccinated sows had slight vascular congestion on the brain meninges, influx of neutrophils, lymphocytes, and shedded epithelial cells in the lung alveolar space, and signs of suppurative bronchopneumonia, hemorrhagic bronchial pneumonia, and lung abscess (Fig. 4a and b). In piglets i.v. challenged by live S. suis, the main lesions (found in the joints, spleen, liver, and kidneys) are associated with synovitis, spleen inflammation, hepatitis, and pneumonia [25]. Dissection of the piglet that died on the third day after challenge showed severe lesions in the lung. In contrast, lesions were not found in any of the piglets from sows in the vaccinated group after dissection, indicating that the enhanced passive immunity against S. suis was sufficient to protect piglets from S. suis challenge.
Prepartum immunization of inactivated S. suis bacterin plus rSao in sows was able to increase serum antigen-specific antibody titers, the concentrations of IFN-γ, IL-4, IL-6 and IL-12 in serum, and provide cross-protection against S. suis infections in the piglets. The protection was characterized by the reduction on lesion score (75 and 81%), bacteremia, and fever. This vaccine and immunization approach may be applied to enhance the protection against both heterologous and homologous S. suis infections in newborn piglets.
Abpb:
Amylase-binding protein B
ANOVA:
BHI:
Brain-heart infusion
BSA:
Bovine serum albumin
CPS:
Capsular polysaccharides
E. coli :
EF:
Extracellular factor
i.m:
i.v.:
Intravenous
IFN-γ:
IgG:
IPTG:
Isopropyl-β-d-thiogalactoside
LPXTG:
Leu-Pro-X-Thr-Gly
Muramidase-released protein
ND50 :
Neutralization dilution 50%
NK cell:
Natural killer cell
Progressive atrophic rhinitis
rsPMT:
Recombinant Pasteurella multocida toxin
S/P:
Sample to positive ratio
Sao:
Surface antigen one
SAS:
Statistical analysis system
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
SsnA:
Surface-anchored DNA-nuclease
T-helper cell
w/o/w:
Water in oil in water
Lun ZR, Wang QP, Chen XG, Li AX, Zhu XQ. Streptococcus suis: an emerging zoonotic pathogen. Lancet Infect Dis. 2007;7(3):201–9.
Hill JE, Gottschalk M, Brousseau R, Harel J, Hemmingsen SM, Goh SH. Biochemical analysis, cpn60 and 16S rDNA sequence data indicate that Streptococcus suis serotypes 32 and 34, isolated from pigs, are Streptococcus orisratti. Vet Microbiol. 2005;107(1–2):63–9.
Higgins R, Gottschalk M. Distribution of Streptococcus suis capsular types in 1998. Can Vet J. 1999;40(4):277.
Chang CJ, Pan CH, Lee SH, Huang TS, Wang C, Jong MH. Analysis of porcine common pathogens in pig herd in Taiwan. Anim Health Res Inst. 2007;42:71–8.
Chu CY, Shu SF, Huang JH, Hsu JP, Xue KR. Rapid detection of Streptococcus suis serotypes and virulent factors in southern Taiwan by multiplex polymerase chain reaction. Taiwan Vet J. 2009;35(2):107–14.
Pallares FJ, Schmitt CS, Roth JA, Evans RB, Kinyon JM, Halbur PG. Evaluation of a ceftiofur-washed whole cell Streptococcus suis bacterin in pigs. Can J Vet Res. 2004;68(3):236–40.
Wisselink HJ, Stockhofe-Zurwieden N, Hilgers LA, Smith HE. Assessment of protective efficacy of live and killed vaccines based on a non-encapsulated mutant of Streptococcus suis serotype 2. Vet Microbiol. 2002;84(1–2):155–68.
Elliott SD, Clifton-Hadley F, Tai J. Streptococcal infection in young pigs. V. An immunogenic polysaccharide from Streptococcus suis type 2 with particular reference to vaccination against streptococcal meningitis in pigs. J Hyg (Lond). 1980;85(2):275–85.
Goyette-Desjardins G, Calzas C, Shiao TC, Neubauer A, Kempker J, Roy R, et al. Protection against Streptococcus suis Serotype 2 infection using a capsular Polysaccharide Glycoconjugate vaccine. Infect Immun. 2016;84(7):2059–75.
Calzas C, Lemire P, Auray G, Gerdts V, Gottschalk M, Segura M. Antibody response specific to the capsular polysaccharide is impaired in Streptococcus suis serotype 2-infected animals. Infect Immun. 2015;83(1):441–53.
Okwumabua O, Abdelmagid O, Chengappa MM. Hybridization analysis of the gene encoding a hemolysin (suilysin) of Streptococcus suis type 2: evidence for the absence of the gene in some isolates. FEMS Microbiol Lett. 1999;181(1):113–21.
Maione D, Margarit I, Rinaudo CD, Masignani V, Mora M, Scarselli M, et al. Identification of a universal group B streptococcus vaccine by multiple genome screen. Science. 2005;309(5731):148–50.
Li Y, Martinez G, Gottschalk M, Lacouture S, Willson P, Dubreuil JD, et al. Identification of a surface protein of Streptococcus suis and evaluation of its immunogenic and protective capacity in pigs. Infect Immun. 2006;74(1):305–12.
Feng Y, Pan X, Sun W, Wang C, Zhang H, Li X, et al. Streptococcus suis enolase functions as a protective antigen displayed on the bacterial cell surface. J Infect Dis. 2009;200(10):1583–92.
Gomez-Gascon L, Cardoso-Toset F, Amarilla PS, Tarradas C, Carrasco L, Olaya-Abril A, et al. A new recombinant SsnA protein combined with aluminum hydroxide protects mouse against Streptococcus suis. Vaccine. 2014;32(51):6992–9.
Huang K, Yuan Z, Li J, Zhang Q, Xu Z, Yan S, et al. Identification and characterisation a surface-associated arginine peptidase in Streptococcus suis serotype 2. Microbiol Res. 2015;170:168–76.
Zhang YM, Shao ZQ, Wang J, Wang L, Li X, Wang C, et al. Prevalent distribution and conservation of Streptococcus suis Lmb protein and its protective capacity against the Chinese highly virulent strain infection. Microbiol Res. 2014;169(5–6):395–401.
Li Y, Gottschalk M, Esgleas M, Lacouture S, Dubreuil JD, Willson P, et al. Immunization with recombinant Sao protein confers protection against Streptococcus suis infection. Clin Vaccine Immunol. 2007;14(8):937–43.
Hsueh KJ, Lee JW, Hou SM, Chen HS, Chang TC, Chu CY. Evaluation on a Streptococcus suis vaccine using recombinant sao-l protein manufactured by bioreactors as the antigen in pigs. Transbound Emerg Dis. 2014;61(6):e35–43.
Liao CM, Huang C, Hsuan SL, Chen ZW, Lee WC, Liu CI, et al. Immunogenicity and efficacy of three recombinant subunit Pasteurella multocida toxin vaccines against progressive atrophic rhinitis in pigs. Vaccine. 2006;24(1):27–35.
Francis MJ, Black L. The effect of vaccination regimen on the transfer of foot and mouth disease antibodies from the sow to her piglets. J Hyg (Lond). 1984;93(1):123–31.
Lipowski A, Drexler C, Pejsak Z. Safety and efficacy of a classical swine fever subunit vaccine in pregnant sows and their offspring. Vet Microbiol. 2000;77(1–2):99–108.
Chu CY, Chen JJ, Chen H. Effect of water-in-oil in-water (W/O/W) oil adjuvant on long-term antibody response of bovine ephemeral fever vaccine Taiwan. Taiwan Vet J. 2007;33(3–4):211–6.
Smith HE, Veenbergen V, van der Velde J, Damman M, Wisselink HJ, Smits MA. The cps genes of Streptococcus suis serotypes 1, 2, and 9: development of rapid serotype-specific PCR assays. J Clin Microbiol. 1999;37(10):3146–52.
Wisselink HJ, Vecht U, Stockhofe-Zurwieden N, Smith HE. Protection of pigs against challenge with virulent Streptococcus suis serotype 2 strains by a muramidase-released protein and extracellular factor vaccine. Vet Rec. 2001;148(15):473–7.
Bourne FJ. The immunoglobulin system of the suckling pig. Proc Nutr Soc. 1973;32(3):205–15.
Nguyen TV, Yuan L, Azevedo MS, Jeong KI, Gonzalez AM, Saif LJ. Transfer of maternal cytokines to suckling piglets: in vivo and in vitro models with implications for immunomodulation of neonatal immunity. Vet Immunol Immunopathol. 2007;117(3–4):236–48.
Zeng R, Spolski R, Finkelstein SE, Oh S, Kovanen PE, Hinrichs CS, et al. Synergy of IL-21 and IL-15 in regulating CD8+ T cell expansion and function. J Exp Med. 2005;201(1):139–48.
Arulanandam BP, Lynch JM, Briles DE, Hollingshead S, Metzger DW. Intranasal vaccination with pneumococcal surface protein A and interleukin-12 augments antibody-mediated opsonization and protective immunity against Streptococcus pneumoniae infection. Infect Immun. 2001;69(11):6718–24.
Sullivan SP, Koutsonanos DG, Del Pilar MM, Lee JW, Zarnitsyn V, Choi SO, et al. Dissolving polymer microneedle patches for influenza vaccination. Nat Med. 2010;16(8):915–20.
Hsieh CS, Macatonia SE, Tripp CS, Wolf SF, O'Garra A, Murphy KM. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science. 1993;260(5107):547–9.
Nathan CF, Murray HW, Wiebe ME, Rubin BY. Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity. J Exp Med. 1983;158(3):670–89.
Zhu Y, Berg M, Fossum C, Magnusson U. Proinflammatory cytokine mRNA expression in mammary tissue of sows following intramammary inoculation with Escherichia coli. Vet Immunol Immunopathol. 2007;116(1–2):98–103.
We thank Dr. Tsung-Chou Chang of the Department of Veterinary Medicine, NPUST, for pathological evaluation of the pigs.
This study was partially supported by the grant from the National Science Council, Taiwan (NSC 99-2324-B-020-001-MY2).
All data were presented in the manuscript.
KJH performed the laboratory work, participated in data analysis and wrote the main parts of the manuscript. WBC and CYC designed the concept of the study, supervised the experiments, revised and submitted the final manuscript. The animal model was performed by YCC. LTC and JWL performed statistical analysis and prepared the final draft of the manuscript. All authors read and approved the final manuscript.
All authors have no financial or personal relationships with other people or organizations that could inappropriately influence or bias their work.
Animal experiments were conducted in a private pig farm in Tainan County, Taiwan, with informed consent from the owner to conduct research. The pigs were selected and randomly assigned to the treatment groups for the vaccine trial. All experimental protocols for animal trials were approved by the Animal Care and Use Committee, National Pingtung University of Science and Technology (NPUST). The experiments were conducted based on the Ethical Rules and Law of NPUST.
Department of Veterinary medicine, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan
Kai-Jen Hsueh
& Wen-Bin Chung
Graduate Institute of Animal Vaccine Technology, College of Veterinary Medicine, National Pingtung University of Science and Technology, 1, Shuehfu Road, Neipu, Pingtung, 91201, Taiwan
, Li-Ting Cheng
, Yao-Chi Chung
& Chun-Yen Chu
Department of Tropical Agriculture and International Cooperation, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan
Jai-Wei Lee
Search for Kai-Jen Hsueh in:
Search for Li-Ting Cheng in:
Search for Jai-Wei Lee in:
Search for Yao-Chi Chung in:
Search for Wen-Bin Chung in:
Search for Chun-Yen Chu in:
Correspondence to Wen-Bin Chung or Chun-Yen Chu.
Hsueh, K., Cheng, L., Lee, J. et al. Immunization with Streptococcus suis bacterin plus recombinant Sao protein in sows conveys passive immunity to their piglets. BMC Vet Res 13, 15 (2016). https://doi.org/10.1186/s12917-016-0937-8
rSao
Passive immunity


text:
Mom & Me & Mom
Read by Maya Angelou
Maya Angelou Random House Audio
The story of Maya Angelou's extraordinary life has been chronicled in her multiple bestselling autobiographies. But now, at last, the legendary author shares the deepest personal story of her life: her relationship with her mother. For the first time, Angelou reveals the triumphs and struggles of being the daughter of Vivian Baxter, an indomitable spirit whose petite size belied her larger-than-life presence—a presence absent during much of Angelou's early life. When her marriage began to crumble, Vivian famously sent three-year-old Maya and her older brother away from their California home to live with their grandmother in Stamps, Arkansas. The subsequent feelings of abandonment stayed with Angelou for years, but their reunion, a decade later, began a story that has never before been told. In Mom & Me & Mom, Angelou dramatizes her years reconciling with the mother she preferred to simply call "Lady," revealing the profound moments that shifted the balance of love and respect between them. Delving into one of her life's most rich, rewarding, and fraught relationships, Mom & Me & Mom explores the healing and love that evolved between the two women over the course of their lives, the love that fostered Maya Angelou's rise from immeasurable depths to reach impossible heights.
Oprah Pick for Best Memoirs of a Generation
A New York Times bestseller
A Los Angeles Times bestseller
An NPR bestseller
A San Francisco Chronicle bestseller
A 2013 Publishers Weekly Listen-Up Award Nominee
A Library Journal bestseller
The story of Maya Angelou's extraordinary life has been chronicled in her multiple bestselling autobiographies. But now, at last, the legendary author shares the deepest personal story of her life: her relationship with her mother.
For the first time, Angelou reveals the triumphs and struggles of being the daughter of Vivian Baxter, an indomitable spirit whose petite size belied her larger-than-life presence—a presence absent during much of Angelou's early life. When her marriage began to crumble, Vivian famously sent three-year-old Maya and her older brother away from their California home to live with their grandmother in Stamps, Arkansas. The subsequent feelings of abandonment stayed with Angelou for years, but their reunion, a decade later, began a story that has never before been told. In Mom & Me & Mom, Angelou dramatizes her years reconciling with the mother she preferred to simply call "Lady," revealing the profound moments that shifted the balance of love and respect between them.
Delving into one of her life's most rich, rewarding, and fraught relationships, Mom & Me & Mom explores the healing and love that evolved between the two women over the course of their lives, the love that fostered Maya Angelou's rise from immeasurable depths to reach impossible heights.
"Angelou's performance captures the prickly early days when thirteen-year-old Maya first meets her mother. We hear Maya's growing respect for Vivian's indomitable spirit as Vivian proves to be Maya's strongest supporter, encouraging her to go after life with 'power and determination.' You won't want to stop listening to Angelou's melodic cadences as she tells of a disastrous visit to her father, of being nearly beaten to death, and of her enviable relationship with her tenacious mother. Each character leaps into full-bodied life, and Angelou's crisp, compressed prose creates unforgettable poetic imagery." —AudioFile
"An evolving portrait of two women learning to understand and respect one another." —Barnes&Noble.com
"Written with her customary eloquence…the lessons and the love presented here will speak to those try to make their way in the world." —Publishers Weekly
"A beautiful tribute to [her mother's] independent, vibrant, and courageous spirit." —Library Journal
"A tightly strung, finely tuned memoir about life with her mother." —Kirkus Reviews
Author Bio: Maya Angelou
Maya Angelou (1928–2014) was a singer, actress, dancer, activist, filmmaker, writer, and mother. In addition to her bestselling autobiographies she wrote several volumes of poetry, including "On the Pulse of the Morning" for the inauguration of President Clinton. She was given a lifetime appointment as Reynolds Professor of American Studies at Wake Forest University in North Carolina, and in 2010 President Barack Obama awarded her the Medal of Freedom, the country's highest civilian honor.