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Q: Cartoon episode where the protagonists encounter a futuristic shopping mall with no exit I vaguely remember seeing an episode of a children's cartoon series on German television in the 90s where the group of protagonists finds a futuristic looking mall. Upon entering it, they are welcomed by some sort of host (I seem to remember it as a hovering robot, but that could be nonsense), and when they turn around, the entrance behind them is gone. None of the shoppers in the mall are aware that "the outside" exists, and they seem confused by or even laugh at the idea of an exit. The episode then revolves around trying to exit the mall and proving to the shoppers that the outside world exists. In the end, I think the ceiling opens and everybody looks at the sky and the sun in awe.
I remember the animation style to be very colorful and typical of 80s and 90s cartoons for children. It was definitely drawn and not computer animated. Other than that, I cannot remember too much about the style, and I'm not entirely sure if it was Japanese-style or western-style, but I'm leaning towards the latter. As mentioned before, I saw it on German TV at some time in the (mid) 90s, and it was professionally dubbed in German.
Despite the somewhat menacing scenario and serious undertones about consumerism, I think the general tone was rather light. It might even be (but I'm really not sure there) that the host apologizes in the end, making the happy ending even more harmonic. I suppose the target audience was something like age 7-12.
I hardly remember anything about the characters. The group of protagonists consisted of around three to five humanoids (probably just humans, but I'm not entirely sure). They were relatively young (but I'm not sure if they were children or teenagers) and I think there were both boys and girls. I'm not sure about their ethnicity, but I think they were (mostly?) white.
Any help in identifying the TV show and the episode is highly appreciated.
PS: I went through this checklist and tried to answer all of its questions as good as I could. If my question is missing any information that this checklist asks for, then this means that I cannot remember anything about it.

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New VLOG posts to come on interview tips and tricks

August 11, 2018

I am currently considering a position at Google, doing machine learning on the core search team. I'm very excited about joining the team. Since it was a pretty long road getting there, look for some new posts in both vlogs and medium.

Interviewing for engineering positions is quite difficult, and it's nerve racking. I worked with a few friends, and that helped me build a lot of perspective about how to pass the interview process.

The Machine Learning Interview Checklist

coding strategy

coding questions

machine learning

system design

Each one of these is important in your algorithm design, theoretical understanding, and the underpinnings of the essential Google interview.

One thing I also appreciated was the insider understanding of the process. Google's process, in particular, exceedingly long and bureaucratic. There was a system in place, people had parts to play, and the procedural grind was setup to be long and arduous.

In a few blog posts, I'll detail what to expect when you're interviewing, when you've finished interviewing, and how to negotiate your salary. Admittedly, I didn't do too hot on the last item there, but by the time you've reached that point, there are tons of resources that are reliable on the web.

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using UnityEngine;
using System.Collections;

public class CameraShake : MonoBehaviour
{
    void Start()
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        iTween.ShakePosition(gameObject, iTween.Hash("amount", Vector3.left/4f, "looptype", iTween.LoopType.loop, "delay", 0f));
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}


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Bratchel, M. E.
Medieval Lucca : And the Evolution of the Renaissance State
The first scholarly study covering the history of both the city and the region of Lucca, from classical antiquity to the end of the fifteenth century - ;Although there are many books in English on the city and state of Lucca, this is the first scholarly study to cover the history of the entire region from classical antiquity to the end of the fifteenth century. At one level, it is an archive-based study of a highly distinctive political community; at another, it is designed as a contribution to current discussions on power-structures, the history of the state, and the differences between city-states and the new territorial states that were
emerging in Italy by the fourteenth century.
There is a rare consensus among historians on the characteristic features of the Italian city-state: essentially the centralization of economic, political, and juridical power on a single city and in a single ruling class. Thus defined, Lucca retained the image of an old-fashioned, old-style city-republic right through until the loss of political independence in 1799. No consensus exists with regard to the defining qualities of the Renaissance state. Was it centralized or de-centralized;
intrusive or non-interventionist? The new regional states were all these things. And the comparison with Lucca is complicated and nuanced as a result.
Lucca ruled over a relatively large city territory, in part a legacy from classical antiquity. Lucca was distinctive in the pervasive power exercised over its territory (largely a legacy of the region's political history in the early and central middle ages). In consequence, the Lucchese state showed a marked continuity in its political organization, and precociousness in its administrative structures. The qualifications relate to practicalities and resources. The coercive powers and
bureaucratic aspirations of any medieval state were distinctly limited, whilst Lucca's capacity for independent action was increasingly circumscribed by the proximity (and territorial enclaves) of more powerful and predatory neighbours. -
Keywords: HISTORY / General HIS000000
Food in Medieval England : Diet and Nutrition
, C. M. Woolgar
You, The People: The United Nations, Transitional Administration, and State-Building
Chesterman, Simon
Medieval Single Women : The Politics of Social Classification in Late Medieval England
Beattie, Cordelia
The Oxford History of Literary Translation in English : Volume 1: To 1550
Ellis, Roger
The Italian Renaissance: Culture and Society in Italy
Burke, Peter
The Ulster Renaissance : Poetry in Belfast 1962-1972
Clark, Heather
The Oxford History of the Irish Book, Volume III : The Irish Book in English, 1550-1800
, Raymond Gillespie
Waterways and Canal-Building in Medieval England
Blair, John
Liberty and Authority in Victorian Britain
Mandler, Peter

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text:
info@teachgiveinspirefridays.com
Home » September 17, 2021
Doors open at 9:30AM
12615 Tribunal Lane
Brunch will be served.
Valet parking is provided.
Covid 19 vaccination required.
THE THEME FOR THE DAY – "FROM STRENGTH TO STRENGTH," will highlight the Power of Women, to own who we are at every stage, to help inspire others to be their best selves, and to connect with and shape future generations. Whether you're a daughter, mother, or grandmother, TGIF has something for you.
Our September event features Maria Olsen (Author, Attorney, Public Speaker, Podcaster), Siobhan Davenport (Author, Philanthropist & CEO, Crittenton Services of Greater Washington) and Meredith Jacobs (Author, Award-winning Journalist & CEO, JWI International).
Maria will TEACH us how to become the best version of ourselves. Siobhan will share her experiences of GIVING back to the community through her leadership and dedication to racial and gender equity for young women, and Meredith will INSPIRE us with her commitment to "Creating, Connecting and Celebrating," family ties and bonds, and community initiatives to keep young women safe.
MARIA OLSEN – Teach Speaker
Author, Attorney, Public Speaker, Podcaster
REFRAMING THE NEXT CHAPTER OF YOUR LIFE
Maria Leonard Olsen is a biracial woman whose parents were forbidden by law to marry in their home state of Maryland in the early 1960s. She is the mother of two children, a lawyer, journalist, and author of two adult nonfiction books and two children's books. She leads writing/empowerment retreats for women, hosts the "Becoming Your Best Version" podcast and is a co-host of the Inside Out radio show on WPFW-fm, 89.3, in Washington, D.C. Her talks on becoming your best version involve overcoming common battles, including divorce, empty-nest syndrome, work/life balance, sexual assault, feeling less than, addictions, anxiety and depression. As a biracial speaker, she has a unique stage presence and ability to connect with wide audiences.
She has written for The Washington Post, Washingtonian, Bethesda Magazine, Parenting, BabyTalk and Washington For Women, and is a contributor to Thrive Global, Up Journey and BreaknLinks. She recently returned to practicing law now that her children are launched. Mommy, Why's Your Skin So Brown? is her first children's book. Healing for Hallie is her second children's book. Not the Cleaver Family-The New Normal in Modern American Family, was released in November 2016. Her latest book, 50 After 50-Reframing the Next Chapter of Your Life, was published in hardcover in 2018, in paperback and on Audible in 2019 and was selected for the National Press Club's Book Fair.
Maria graduated from Boston College and the University of Virginia School of Law, and served in the Clinton Administration's Justice Department.
SIOBHAN DAVENPORT – Give Speaker
Author, Philanthropist, President and CEO, Crittenton Services
CHAMPION FOR WOMEN AND GIRLS. EMPOWERING YOUTH TO ACHIVE THEIR DREAMS
Siobhan Davenport is author and President & CEO of Crittenton Services of Greater Washington– a nonprofit that partners with schools to help girls from the 6th to 12th grades overcome obstacles, make positive choices, and achieve their goals.
As the daughter of teen parents, Siobhan knows a thing or two about overcoming the odds. Personal willpower is important, but Siobhan believes it is imperative to have a community that fosters success. She has dedicated her career to racial and gender equity by working to give young girls the tools to overcome challenges and the community and support systems that enable them to thrive.
In her current role as President & CEO of Crittenton Services of Greater Washington (CSGW), she is laser-focused on closing equity and achievement gaps for teen girls, particularly those of color, in the DC Metropolitan area. Crittenton partners with schools to run multi-year programs that drive youth social, emotional, and academic success.
Prior to serving as CEO, she was CSGW's Director of Strategic Partnerships and built collaborations between schools, nonprofits, and corporations to better meet Crittenton girls' challenges. Siobhan also launched the Declare Equity for Girls: It's Time campaign; a landmark initiative that included focus groups and exposed the major barriers to personal and academic success. She was also the former executive director of the Rocksprings Foundation from 2002 to 2018 where she oversaw the foundation's impact by investing in economic and educational opportunities for low-income families and youth.
Additionally, Siobhan is an author, commissioner for the Maryland Governor's Commission on Service and Volunteerism and sits on the Advisory Council for Ascend at the Aspen Institute. Siobhan was also crowned Mrs. Maryland in 2008. Aspiring to be a role model for her own daughter and all young girls, Siobhan is a living demonstration of following your dreams and building others up around you.
MEREDITH JACOBS – Inspire Speaker
Author, Award-winning Journalist, CEO, JWI International
CREATE, CONNECT, CELEBRATE
Meredith Jacobs, JWI's CEO, is an award-winning journalist and former editor-in-chief of Washington Jewish Week. She is the author of The Modern Jewish Mom's Guide to Shabbat: Connect and Celebrate—Bring Your Family Together with the Friday Night Meal (HarperCollins) and co-author, with her daughter Sofie, of the bestselling series of interactive journals, Just Between Us (Chronicle Books). Prior to joining JWI, she founded ModernJewishMom.com, the first Jewish parenting website (now part of Kveller), and was the host of WYPR radio's Connecting Family and The Jewish Channel's holiday specials, Modern Jewish Mom.
Jacobs assumed the role of CEO after serving as JWI's chief operating officer for six years; in that role she managed communications, branding, messaging, and development. She has shepherded numerous JWI leadership initiatives, including the Jewish Communal Women's Leadership Project; Men As Allies: Leading Equitable Workplaces ; the Women's Impact Network and the Young Women's Impact Network. Jacobs also works closely with JWI's philanthropic partners, Sigma Delta Tau national sorority and Zeta Beta Tau national fraternity, developing initiatives like the award-winning Green Light, Go! and Girls Achieve GrΣΔΤness. A sought-after speaker, moderator and writer, her opinion pieces appear frequently in outlets such as JTA, eJewishPhilanthropy, and Washington Jewish Week. The Jerusalem Post named Jacobs one of the 50 most influential Jews of 2020.
She lives in suburban D.C. with her husband and their giant pandemic puppy, Babka. Her greatest achievements are her children, Sofie, a marketing strategist, and Jules, a Navy officer.
TGIF is a women's speaker series that will feature quarterly Friday parlor style meetings with thought leaders and industry movers and shakers, designed to empower and ignite creativity and possibilities for women in our community.
tgif2022.eventbrite.com
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Genome Study Predicts DNA of the Whole of Iceland
Complete the form below and we will email you a PDF version of "Genome Study Predicts DNA of the Whole of Iceland"
The CEO of an Icelandic gene-hunting company says he is able to identify everyone from that country who has a deadly cancer risk, but has been unable to warn people of the danger because of ethics rules governing DNA research.
The company, DeCode Genetics, based in Reykjavík, says it has collected full DNA sequences on 10,000 individuals. And because people on the island are closely related, DeCode says it can now also extrapolate to accurately guess the DNA makeup of nearly all other 320,000 citizens of that country, including those who never participated in its studies.
That's raising complex medical and ethical issues about whether DeCode, which is owned by the U.S. biotechnology company Amgen, will be able to inform members of the public if they are at risk for fatal diseases.
Kári Stefánsson, the doctor who is founder and CEO of DeCode, says he is worried about mutations in a gene called BRCA2 that convey a sharply increased risk of breast and ovarian cancers. DeCode's data can now identify about 2,000 people with the gene mutation across Iceland's population, and Stefánsson said that the company has been in negotiations with health authorities about whether to alert them.
"We could save these people from dying prematurely, but we are not, because we as a society haven't agreed on that," says Stefánsson. "I personally think that not saving people with these mutations is a crime. This is an enormous risk to a large number of people."
The Icelandic Ministry of Welfare said a special committee had been formed to regulate such "incidental" findings and would propose regulations by the end of the year.
The technique used by DeCode to predict people's genes offers clues to the future of so-called precision medicine in other countries, including the U.S., where this year President Barack Obama called for researchers to assemble a giant database of one million people (see "U.S to Develop DNA Study of One Million People"). A large enough U.S. database could also be used to infer genes of people whether or not they had joined it, says Stefánsson, and could raise similar questions about whether and how to report health hazards to the public.
"This technique can be applied to any population," says Myles Axton, chief editor of Nature Genetics, the journal in which DeCode today presented some of its findings. He said the tiny island's detailed genealogical records are why "it was achieved first in Iceland."
Various legal and ethical obstacles currently prevent DeCode from warning people who are at risk. Volunteers who signed up for DeCode's studies were promised anonymity, and also that they wouldn't learn of research findings. Bioethicists recognize that people have a right "not to know" of genetic hazards, which means they can't simply be told.
"The rule is that you can only use and expose genetic data if you have the permission from the individual in question," says Gísli Pálsson, an anthropologist at the University of Iceland. "But this is beyond informed consent. People are not even in the studies, they haven't submitted any consent or even a sample, yet the company claims to have knowledge about these people and that there is a health risk."
Pálsson says traditional notions of medical ethics are now in open conflict with the aims and capabilities of precision medicine. He believes such standards will need to be adjusted in fundamental ways in the future, so that more weight is given to public health benefits over individual privacy rights.
The life expectancy of women with the BRAC2 mutation is 12 years less than for women without it because 86 percent of those who have it will develop cancer. Men are also affected because the mutation raises the chance of prostate cancer. Stefansson says many of those deaths could be avoided by preventive surgery, like a mastectomy.
"We could in Iceland, at the push of a button, find all women with a mutation in the BRCA2 gene," says Stefánsson. "It is one tiny little example of what you can do. You can use this in preventive medicine like never before."
DeCode's data might also be able to predict who in the population is at elevated relative risk for Alzheimer's disease or who has an undiagnosed learning disability.
DeCode was started in 1996 with the idea of linking DNA research to Iceland's national health system (see "Population Inc."). The country's situation is unique because of its carefully kept genealogical records, which go back to the Ninth Century, when the island was settled. That also means most Icelanders are at least distant cousins.
DeCode has generated rough gene maps, called genotypes, on about 150,000 Icelanders. More recently, advances in DNA sequencing technology have allowed it to obtain complete genome sequences of about 10,000 people, Stefánsson says, or about one in 30 Icelanders. Results from the first 2,636 full genomes were reported by the company in Nature Genetics.
Combined with genealogical tables, these data are what allow DeCode to "impute" the genomes of nearly everyone born of Icelandic parents. That is because once the full genome of some people in a family tree is known, the genes of the others can be inferred.
Sean Harper, head of R&D at Amgen, calls DeCode's ability to guess people's DNA a "conundrum." He says it is not an unfamiliar problem to geneticists, however, since routine DNA tests of one person often reveal information about close relatives. But it has never before occurred at the level of an entire nation.
"It's a gray area from a bioethics perspective, but we would be inclined to provide the data or at least a notice," says Harper. He says Amgen, which uses gene information from DeCode to guide its drug research, had no plans to charge anyone for the risk information it has developed. "It would not be appropriate [to] try to make it a commercial process," he says.
The U.S. lacks a similar national database, although some private companies, like 23andMe and Ancestry.com, have generated rough gene maps of several million people altogether. The National Institutes of Health is planning to spend millions in the coming years on accumulating full genome data on tens of thousands of people.
That means the entire world will soon confront the same types of ethical dilemmas that Iceland now faces, says Pálsson. "Do you have the right to fiddle with people's lives on a massive scale? You can tell your neighbor, 'You are smoking too much.' But it's another thing to approach 1,000 people and say, 'You have the BRCA2 mutation,' " he says.


Follow Megan on Twitter, Instagram and Facebook!
Megan DeSales Colarossi co-anchors Good Day LA between 9am and 10am along with Vanessa Borge.
Megan was born and raised in Fullerton, California, where she graduated from Sunny Hills High School in 1993. Before calling New York City home for almost 20 years, she was first drawn South to Vanderbilt University in Nashville, Tennessee where she received her B.A. in Human and Organizational Development and Communications.
In 1997, Colarossi signed on with CNN where she worked behind the camera at the cable networks Los Angeles, New York and Atlanta bureaus. Quickly rising from associate producer to senior producer, Megan produced various live and long format programs including CNN Newsstand: Entertainment Weekly, CNNfn Entrepreneurs Only, and Headline News Primetime Live.
After a little over 4 years at CNN and hundreds of breaking news events later, she followed her passion for music and headed to MTV News in 2001. As the assistant managing editor and senior producer for the music channel, she interviewed everyone from rock stars to presidents.
For the next six years, Megan traveled around the world for the music channel writing and producing award-winning pro-social programs about the war in Iraq, the tsunami in Indonesia, and the poverty crisis in Africa.
After many years in television news, Colarossi decided to dip her hand in other career endeavors. In 2006, she co-wrote a fiction novel called Spooning (Random House). In 2007, she followed her love of indoor cycling and helmed marketing and PR for a relatively unknown NYC spinning studio called SoulCycle. And in 2009, she helped a close friend launch a non-profit, The R.E.S.T. Initiative, which brought free massages to thousands of cancer patients at NYU Clinical Cancer Center in New York City.
In early 2013, Megan's career path took yet another dramatic turn and she came back to the world of television, but this time in front of the camera. Her dear friend of almost 20 years and Today Show co-host, Willie Geist, convinced her to come on the morning show with him in a regular segment called TODAY's Friendly Advice to offer millions of viewers her spot on and witty advice when it comes to life's little dilemmas and sticky situations.
From "How do you tell a friend they have bad breath?" to "Is it ok to be friends with an ex?" Willie and Meg tackle issues that come up in our everyday lives. The premise is simple: Sometimes it's better to get advice from a friend versus an expert!
Willie and Megan also hosted a TODAY.com digital series called Parental Guidance. The dynamic duo, who are also parents, tackled common dilemmas that moms and dads face every single day. From lice and leaky diapers to cussing and curfews, Willie and Meg offered simple solutions that would hopefully make parents lives a little bit easier.
With a dash of humor and a dose of good'ole fashioned ingenuity, no tantrum was too big or too small for this witty pair to handle!
From 2015-2016, Megan was a daily panelist on the nationally syndicated talk show The Meredith Vieira Show on NBC. Everyday Meredith and her fellow co-panelists would dish about What's Hot Now!. From breaking news and current events to parenting and pop culture, they covered everything and anything. Let's just say no topic was off limits with this dynamic group!
In the Fall of 2016, Megan co-hosted The Way Home, a weekly, lifestyle and home makeover series on the A&E Network. The show was a go-to source for décor, design, tips, tricks, hacks, and how-tos for the home and family.
However, Megan's most proud accomplishment has always been being a mother to two little girls, Hudson and Quinn.


6" x 8-1/2" x 1/2"
This extra large Blue Guards photo crystal has a rectangular central panel, guarded on the sides by two blue crystals. The blue crystals are slightly thicker than the central panel, providing extra elegance to the overall design. This laser photo crystal is popular among corporate customers for various occasions.
The Blue Guards photo crystal has a rectangular center panel, guarded on the sides by two blue crystals.
The blue crystals are slightly thicker than the center panel, providing extra elegance to the overall design.
Size: 6" width x 8-1/2" height x 1/2" thickness.
This photo crystal with attached crystal base is a very popular personalized photo crystal gift for many occasions.


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Home/News/Punjab Protection Of The Foundation Of Islam Act: A Step Towards Religious Harmony Or An Attempt To Suppress Free Thinking?
Punjab Protection Of The Foundation Of Islam Act: A Step Towards Religious Harmony Or An Attempt To Suppress Free Thinking?
Punjab Protection Of The Foundation Of Islam Act - News Mag
The Punjab Assembly of Pakistan has recently passed a law banning all kinds of books or other publications which include 'Allah, the Prophet of Islam, Prophets of other religions, Holy Books, Righteous Caliphs, Companions There will be no insulting, critical or derogatory material about angels, Ahlul Bayt.
It has been named as Punjab Protection of Foundation of Islam Act 2020 and according to the law, wherever the name of the last prophet of Islam will be written in the books, first Khatam-un-Nabiyyin and then in Arabic script, peace be upon him. It will be necessary to write.
The Punjab government is calling the passage of the law a "historic step" and Punjab Chief Minister Usman Bazdar has said in a message that "the implementation of this law has led to hate speech about religions (especially Islam) in various books." Content will be prevented.
However, the Human Rights Commission of Pakistan (HRCP) has "warned the Punjab Assembly that it is inciting prejudice".
A large part of this law deals with restrictions that have nothing to do with Islam or religion.
The new law stipulates that any material that "threatens the ideology of Pakistan or the sovereignty, integrity and security of Pakistan" will be banned.
At the same time, it has been said that such material will not be published "which is against moral values and culture or promotes obscenity or pornography etc."
"The Naked Athena": A New Symbol Against Police Brutality In The United States
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All books and materials that promote terrorism or terrorists and extremism, etc., or that pose a threat to sectarianism, violence and hatred, and the deteriorating law and order situation are also banned.
The Punjab Information and Broadcasting Corporation has been given wide powers to scrutinize books and to allow or not to publish published materials, including raids on printing presses, seizure of materials, investigation and scrutiny of financial accounts. Options like
What Is In The New Law?
Under the new law, there are two different types of punishments. A punishment will only be given for violating section 3F of the law, which will be under the current criminal code.
Section 3F of the Punjab Foundation for the Protection of the Foundations of Islam Act is the only section which deals with Islam, which makes it mandatory to use Islamic titles with the names of holy persons.
The law provides for imprisonment for up to five years and a fine of up to Rs. 500,000.
Sub-sections 5 to 11 of section 8 of the law repeat the terms of section 3F once again, but this section refers to the rules and regulations for national and international books.
The HRCP statement also said that "there is ample reason to expect that Section 3F of the new law will be used to target religious minorities and sects."
What Powers Have Been Given To The Information And Broadcasting Corporation?
The Directorate General of Public Relations, Information and Broadcasting Department of the Punjab Government has been empowered in the new law to:
After raiding and inspecting any printing press and bookstore, any book can be confiscated with reference to any material whether it has just been published or not.
He will be able to determine any action or omission in this regard by investigating and inquiring
He will be able to check the records and accounts of the printing press on the spot or compel him to present those records in the office of his appointed representative.
He will be able to transfer his powers to any officer
However, the powers of the DGPR do not end here. In the new law, it will be central to the implementation of almost all orders.
According to the powers given to the DGPR to regulate national and foreign books in Section 8 of the Act:
No person shall publish, publish or import any book or material which has been previously published or broadcast in Pakistan or abroad without the permission of DGPR.
To publish or publish any book or material, the DGPR has to apply on a form
Under section one of the law, if the book or material is against the national interest, civilization, religious and sectarian harmony, the DGPR will refuse to allow it.
The DGPR will be required to send any book or material related to religion to the Muttahida Ulema Board under sections one and three of the law.
The Human Rights Commission of Pakistan has termed it unacceptable for the DGPR to be empowered to decide whether any material is objectionable or not.
Will The Restrictions Be Limited To Books Only?
The definition of 'book' in the new law is not limited to books. According to this definition, wherever the word book is used in law, it means "every skin or part of the skin and pamphlet in any language."
'Charts or plans based on music, logos, maps that have been individually reprinted or reprinted or reproduced or made on lithographs or any other soft format on discs and similar digital and electronic devices I was made. '
However, they will not include newspapers, magazines, journals, magazines, newsletters and textbooks.
Books Will Be Accounted For
Under the law, the DGPR will create a catalog that will be based on the books it receives under the law and will include all the facts about the book. Each book will be assigned a unique number.
The books will be printed only by registered printers and each book publisher will be required to submit a memorandum of printed books to the designated officer every three months.
Similarly, the importer of books will be required to submit a memorandum of imported books to the concerned officer within 15 days of import.
According to the HRCP statement, "they are concerned that such measures will restrict not only freedom of expression but also freedom of thought, conscience and religion."
'Power Is Always Afraid Of The Book'
A statement issued by HRCP Chairperson Dr. Mehdi Hassan said that in Pakistan, Section 505 (2) of the Criminal Code and Section 8 of the Anti-Terrorism Act are already against racial and sectarian hatred. There are penalties.
"The intent of this new legislation is like a stereotype of burning books."
Speaking to the BBC, News Mag Harris Khaliq, HRCP general secretary and poet and intellectual, said such measures were nothing new in Pakistan but their results had never been good.
As he puts it, "Power is not afraid of another power, but power is always afraid of books, music or literature that is revolutionary."
He said that the far-reaching consequences of such measures are crushed by the wheel of history but its immediate consequences create problems for the society.
Why Did The Government Want To Bring This Law?
The Punjab government says the law was needed to prevent incidents such as insulting religion and holy figures. Speaker Punjab Assembly Chaudhry Pervaiz Elahi while approving the law said that in the past such incidents have been pointed out by the members of the Assembly in the form of resolutions.
"These resolutions were also passed and now we have legislated on it. Hopefully, after this law, religious and sectarian aggression will decrease."
He suggested that the federation and other provinces should follow Punjab's example in this regard.
'Banning Books Is Not The Answer'
Regarding the ban on books, historian and author Dr. Mubarak Ali said, "People should have the right to decide for themselves whether to read a book or not or reject it." Banning a book is not the answer.
He said that various terms used in the law are such that they have not been defined and cannot be defined. In this way they can be used to achieve the desired results.
Dr. Mubarak said that the ban on books would stop the process of creation in Pakistan which was already suffering from problems and people were not writing books.
"It's basically an attempt to stop the younger generation from reading modern and progressive literature," he said. Do they want to read only fiction books?
Writer and playwright Amna Mufti told the BBC, that no new literature was being created in Pakistan anyway. 'So who do they want to scare? They are basically afraid of the digital space and want to control it.
"The Naked Athena": A New Symbol Against Police Brutality In The United States
Harris Rauf Joins National Squad In England


Revise ´Did´ question form and short answers. Arrange class in pairs. Give pupils their section of the worksheet. Ask them to question their Partner to complete the chart.
A short quiz about the touching and funny film : Little Miss Sunshine.


Congratulations to RSE Fellow Alison Phipps OBE, UNESCO Chair of Refugee Integration through Languages and the Arts, and Professor of Languages and Intercultural Studies at the University of Glasgow on being elected today to the Fellowship of the Academy of Social Sciences. Alison is a leading researcher and public figure in conflict transformation, sanctuary and asylum seeking, as well as education and languages for peace and non-violence.
Congratulations also to Young Academy of Scotland Member and Co-Chair, Nasar Meer, Professor of Race, Identity and Citizenship at the University of Edinburgh on his election to the Academy. Nasar is a leading scholar in the study of comparative citizenship, including multiculturalism, interculturalism and integration with focuses on Muslim experience and Scottish understanding of nationhood and race.


text:
Q: Making a bordered box in C with a movable character I am having a couple issues, I am learning C throug the Zed Shaw Book, and I thought I would take a small break and try and put what I have learned so far to the test. But then I got stuck.
The first issue is Creating an empty box. I cannot seem to do this. I Can Create a box, missing the bottom part but it is filled in by '*', if I just do '' then the c compiler states: create_border.c:12:14: error: expected expression
The second issue, is I dont know how to connect to the keyboard so that when I press Up, Down, Left, Right a character, @ in this case, moves around and stops when it his an edge.
All I have so far is:
#include <stdio.h>

#define MAX_Y 6
#define MAX_X 6

void print_board(char board[MAX_Y][MAX_X]);

int main(int argc, char *argv[]) {

    char exampleBoard[MAX_Y][MAX_X] = {
        {'x','-','-','-','-','x'},
        {'|','*','*','*','*','|'},
        {'|','*','*','*','*','|'},
        {'|','*','*','*','*','|'},
        {'|','*','*','*','*','|'},
        {'x','-','-','-','-','x'}
    };

    print_board(exampleBoard);

    return 0;
}

void print_board(char board[MAX_Y][MAX_X]) {

    int row = 0;
    int col = 0;

    for(row = 0; row < MAX_Y; row++) {
        for(col = 0; col < MAX_X; col++) {
            printf("%c", board[row][col]);
        }

        printf("\n");
    }

    printf("\n\n");
}

All this prints out is:
x----x
|****|
|****|
|****|
|****|

So heres what I expect:

*

*An empty box, with no '*' and a @ character any where in the box.

*Ability to move @ and have it stop when it reaches a border.
Any help is much appreciated.

A: In a terminal, an "empty case" is a space character. You can write it with ' '  instead of ''.
Then, for reading keyboard, you can make it with read syscall directly from your terminal. But the default configuration of your terminal will force you to press enter between each key to make your read works.
NOTE: You could make this kind of ASCII-programming simplier using a library like ncurses.


text:
<?php
/**
 * @link       http://superdweebie.com
 * @package    Sds
 * @license    MIT
 */
namespace Sds\DoctrineExtensions\Annotation\Annotations\Rest;

use Doctrine\Common\Annotations\Annotation;

/**
 *
 * @since   1.0
 * @author  Tim Roediger <superdweebie@gmail.com>
 *
 * @Annotation
 * @Target({"CLASS"})
 */
final class Cache extends Annotation
{
    const event = 'annotationRestCache';

    public $public = false;

    public $private = false;

    public $noCache = false;

    public $maxAge = false;
}


text:
OtherDRUG METABOLISM AND DISPOSITION
Purification and Characterization of Heterologously Expressed Mouse CYP2A5 and CYP2G1: Role in Metabolic Activation of Acetaminophen and 2,6-Dichlorobenzonitrile in Mouse Olfactory Mucosal Microsomes
Jun Gu, Qing-Yu Zhang, Mary Beth Genter, Thomas W. Lipinskas, Masahiko Negishi, Daniel W. Nebert and Xinxin Ding
Journal of Pharmacology and Experimental Therapeutics June 1998, 285 (3) 1287-1295;
Qing-Yu Zhang
Mary Beth Genter
Thomas W. Lipinskas
Masahiko Negishi
Daniel W. Nebert
Xinxin Ding
The metabolic activation of two known olfactory mucosal (OM) toxicants, acetaminophen (AP) and 2,6-dichlorobenzonitrile (DCBN), was examined with mouse liver and OM microsomes and purified, heterologously expressed mouse CYP2A5 and CYP2G1. In reconstituted systems, both isoforms were active in metabolizing DCBN and AP to metabolites that formed protein adducts. The formation of DCBN- or AP-protein adducts and other AP metabolites, including 3-hydroxy-AP and, in the presence of glutathione, AP-glutathione conjugate, was also detected in OM microsomal reactions and to a much greater extent than in liver microsomes. Evidence was obtained that CYP2A5 and CYP2G1 play major roles in mouse OM microsomal metabolic activation of DCBN and AP. Immunoblot analysis indicated that CYP2A5 and CYP2G1 are abundant P450 isoforms in OM microsomes. OM microsomal AP and DCBN metabolic activation was inhibited by 5- and 8-methoxsalen, which inhibit both CYP2A5 and CYP2G1, and by an inhibitory anti-CYP2A5 antibody that also inhibits CYP2G1. In addition, the roles of CYP1A2 and CYP2E1 in the OM bioactivation of AP and DCBN were ruled out by comparing activities of acetone-treated mice or Cyp1a2(−/−) mice with those of control mice. Thus, CYP2A5 and CYP2G1 may both contribute to the known OM-selective toxicity of AP and DCBN. Further analysis of the kinetics of AP and DCBN metabolism by the purified P450s suggested that CYP2A5 may play a greater role in OM microsomal metabolism of AP, whereas their relative roles in DCBN metabolism may be dose dependent, with CYP2G1 playing more important roles at low substrate concentrations.
Numerous environmental chemicals are known to cause tissue-selective toxicity in the olfactory mucosa in rodents (Bonnefoi et al., 1991; Dahl and Hadley, 1991; Genter et al., 1992; Gu et al., 1997). However, little is known about whether these compounds are also toxic in human nasal mucosa. To better predict the potential toxicity of environmental chemicals in humans and to determine the mechanism of tissue-selective toxicity, we sought to identify the enzymes responsible for their metabolic activation in the animal models exhibiting toxicity.
In mice, several cytochrome P450 [P450; the nomenclature used in this report is according to Nelson et al. (1996)] isoforms have been detected in olfactory mucosa, including CYP1A2, CYP2A5, CYP2E1, CYP2G1 and one or more isoforms of the CYP3A subfamily (Hua et al., 1997; Su et al., 1996; Guet al., 1997; Genter et al., 1998). Of these, CYP2G1 is detected only in the olfactory mucosa (Hua et al., 1996), whereas CYP2A5 is expressed in the olfactory mucosa at levels much higher than in liver, kidney and lung (Su et al., 1996). Heterologously expressed mouse CYP2G1 is active toward testosterone and progesterone (Hua et al., 1997), which is consistent with findings with rabbit CYP2G1 (Ding and Coon, 1994). CYP2A5 is known to be the major coumarin 7-hydroxylase in mouse liver (Negishi et al., 1989) and also has activity toward other foreign chemicals, such as aflatoxin B1 and N-nitrosodiethylamine (Camus et al., 1993; Pelkonen et al., 1994).
In the present study, the roles were examined of CYP2A5 and CYP2G1 in the metabolic activation of two known OM toxicants, DCBN and AP. DCBN is a herbicide used for weed control. AP is a widely used therapeutic agent. Both DCBN and AP have been shown to cause OM toxicity in mice (Jeffery and Haschek, 1988; Brittebo, 1993; Deamer et al., 1994). DCBN does not cause toxicity in liver at the doses that cause OM toxicity (Brandt et al., 1990), whereas AP causes liver and renal toxicity in addition to OM toxicity (Placke et al., 1987; Jeffery and Haschek, 1988).
Metabolic activation of DCBN and AP was studied using purified P450s in reconstituted systems and in liver and OM microsomal preparations. Heterologously expressed mouse CYP2A5 and CYP2G1 were obtained and purified to electrophoretic homogeneity. Antibodies, generated with purified CYP2A5, and chemical inhibitors were used to demonstrate the prominent roles of the two isoforms in OM microsomal metabolism. In addition, the potential roles of CYP1A2 and CYP2E1 were ruled out in experiments with animals lacking the Cyp1a2 gene (Lianget al., 1996) or mice with elevated CYP2E1 levels after acetone treatment. Furthermore, to determine the relative importance of CYP2A5 and CYP2G1 in metabolic activation, the kinetics were examined of AP and DCBN metabolism by the purified P450s. Our results indicate that although both CYP2A5 and CYP2G1 are active toward DCBN and AP, CYP2A5 may play a greater role in OM microsomal metabolism of AP, whereas the relative roles of CYP2A5 and CYP2G1 in DCBN metabolism may be dose dependent, with CYP2G1 playing a more important role at low substrate concentrations.
Heterologous expression and purification of mouse CYP2A5 and CYP2G1.
Baculovirus-mediated heterologous expression of mouse CYP2G1 in insect cells has recently been achieved (Hua et al., 1997). The bacterial expression vector for CYP2A5 was constructed as described previously for CYP2A4 (Sueyoshi et al., 1995). Escherichia coli cells expressing mouse CYP2A5 were cultured in 1 liter of LB medium containing 100 μg ampicillin, 0.2% glucose and 0.5 mM δ-aminolevulinic acid. The cells were incubated in an orbital shaker at 37°C for 4 hr at 225 rpm before addition of isopropyl-β-d-thiogalactoside (Stratagene, La Jolla, CA) to a final concentration of 0.5 mM and were collected after a 48-hr incubation at 25°C and at 150 rpm. The cell pellet from 1 liter of culture was washed in 40 ml of 20 mM potassium phosphate buffer, pH 7.4, containing 50 mM KCl and 5 mM EDTA and resuspended in 2 vol of the same buffer. The cells were lysed by three passes through a French Pressure Cell (SLM) using the high setting and a gauge reading of 600 to 700. Cell debris was removed by centrifugation at 9500 rpm for 10 min in a JA 20 rotor, and the supernatant was centrifuged again at 35,000 rpm in a Ti 45 rotor for 1 hr to obtain the membrane fraction. The membrane pellet was washed with 50 mM Tris-HCl buffer, pH 7.4, containing 0.8 M KCl and 1 mM EDTA, and resuspended in 4 ml of 10 mM Tris-acetate buffer, pH 7.4, containing 1 mM EDTA and 20% glycerol, with a 15-ml Dounce all-glass homogenizer. The membrane fractions from four 1-liter cultures were combined and solubilized by resuspending in 0.5% (w/v) Tergitol NP-10 and 1% (w/v) sodium cholate in 0.1 M Tris-acetate buffer, pH 7.4, containing 0.15 M KCl and 1 mM EDTA, with continuous mixing at 4°C for 2 hr. After removal of insoluble material by centrifugation, the supernatant was dialyzed overnight against 10 mM phosphate buffer, pH 7.7, containing 1 mM EDTA, 0.5% NP-10 and 20% glycerol (buffer A) and then loaded on a 50-ml DEAE-Sepharose column (Sigma Chemical, St. Louis, MO) previously equilibrated with buffer A. CYP2A5 was recovered in the unbound fraction, which was directly applied to a 10-ml HTP column (BioRad, Hercules, CA) previously equilibrated with 10 mM phosphate buffer, pH 7.4, containing 0.5% NP-10 and 20% glycerol (buffer B). After washing with 10-column vol of buffer B, the HTP column was eluted with a linear gradient of potassium phosphate (10–300 mM, pH 7.4) in buffer B. A homogeneous preparation of CYP2A5 was recovered at ∼180 mM phosphate buffer, which after dialysis overnight against buffer B, was applied to a second HTP column (5 ml) to remove Tergitol NP-10 as described previously (Ding and Coon, 1988). The bound CYP2A5 was eluted with 500 mM potassium phosphate buffer, pH 7.4, containing 0.5% sodium cholate and 20% glycerol. The final CYP2A5 preparation was dialyzed twice overnight against 2 liters of detergent-free buffer (50 mM potassium phosphate, pH 7.4, containing 20% glycerol and 0.1 mM EDTA) and stored at −30°C.
SF9 cell microsomes containing mouse CYP2G1 were solubilized using the same protocol as described for the solubilization of CYP2A5 and were fractionated by PEG precipitation (Coon et al., 1978). The resulting 6% PEG supernatant was then diluted (1:2) with 20% glycerol, and a 10% NP-10 stock solution was added to a final concentration of 0.5%. The preparation (∼80 nmol total P450) was loaded onto a 30-ml HTP column previously equilibrated with buffer B and eluted with a potassium phosphate buffer gradient (10–300 mM) as described above. CYP2G1, which was recovered at ∼150 mM phosphate, was dialyzed overnight with 10 mM phosphate buffer, pH 6.4, containing 1 mM EDTA, 0.5% NP-10 and 20% glycerol (buffer C) and loaded onto an 8-ml S-Sepharose column (Sigma) previously equilibrated with buffer C. The column was washed with buffer C and eluted with a pH gradient from 6.4 (buffer C) to 7.4 (buffer B). A homogeneous preparation of CYP2G1 was recovered at pH ∼7.0, which was dialyzed overnight with 10 mM phosphate buffer, pH 7.4, containing 0.2% NP-10 and 20% glycerol and applied to a HTP column (3 ml) to remove Tergitol NP-10. The bound CYP2G1 was eluted with 500 mM potassium phosphate buffer, pH 7.4, containing 0.1% cholate, 0.1 mM EDTA and 20% glycerol. The final CYP2G1 preparation was dialyzed against detergent-free buffer as described above for the purification of CYP2A5.
Preparation of anti-CYP2A5 antibody and immunoblot analysis.
Polyclonal antibodies to CYP2A5 were prepared using purified CYP2A5 in two female Flemish-Giant Chinchilla rabbits using a previously described protocol (Ding and Coon, 1990). IgG fraction was prepared from rabbit serum according to McKinney and Parkinson (1987). Immunoblot analysis was performed with an ECL kit from Amersham as described previously (Ding and Coon, 1990). The source of a monoclonal antibody to rat CYP2E1 (mAb 1–98-1) and preparation of polyclonal antibodies to rabbit CYP2A10/11 and CYP2G1 have been described in earlier studies (Park et al., 1986; Ding and Coon, 1990;Ding et al., 1991).
Determination of catalytic activities.
Formation of DCBN-protein adduct and GSH conjugates of DCBN epoxide (GS-DCBN) was assayed as described recently (Ding et al., 1996) with use of 2,6-[ring-14C]DCBN (16.7 Ci/mol; Sigma) as a substrate. The contents of reaction mixtures and the incubation conditions are indicated in the legends to tables and figures. Protein adducts were precipitated with ice-cold acetone, washed repeatedly with sodium dodecyl sulfate (1%) to remove nonspecific binding, dissolved in 1 M sodium hydroxide and analyzed by liquid scintillation counting. The radioactivity in the washed precipitates was compared with that of the total reaction mixture for calculation of the rate of adduct formation. Formation of GS-DCBN was determined by radiometric HPLC as described previously (Ding et al., 1996).
AP metabolism was assayed essentially according to Morgan et al. (1983) with some modifications in the contents of reaction mixtures as indicated in the legends to tables and figures. Reactions were initiated by the addition of NADPH to a final volume of 250 μl and were terminated after incubation at 37°C for up to 10 min by the addition of 125 μl of 3 M perchloric acid. HPLC analyses of AP metabolites were performed with a Waters μBondapak C18 column according to Harvison et al. (1988), with a radiometric HPLC system as described recently (Ding et al., 1996). Metabolites were detected by absorbance at 250 nm and by radioactivity measurements with14C-AP as the substrate. Metabolites of AP were identified on the basis of comigration with standards generated in enzymatic reactions with rat CYP1A1 in a reconstituted system (data not shown), which was previously shown to metabolize AP to 3-OH-AP and GS-AP in the presence of added GSH (Harvison et al., 1988). The retention times of the two metabolites were similar to the ones reported earlier using the same chromatographic conditions (Harvisonet al., 1988). Formation of GS-AP was also confirmed by its detection by radioactivity measurement in experiments with 3H-GSH and unlabeled AP (data not given). Formation of AP-protein adduct was assayed as described above for DCBN metabolism with use of [ring-UL-14C]-AP (6.3 Ci/mol, from Sigma) as a substrate.
Quantification of 3-OH-AP and GS-AP was initially carried out by measuring radioactivity of individual peaks with an online radioactivity detector, and the amounts were calculated on the basis of percent recovery of total radioactivity. Subsequently, the ratios of UV absorbance at 250 nm to radioactivity of the two metabolites and that of AP were calculated by plotting the integrated peak areas from UV detector vs. those from radioactivity detector using samples from >10 experiments. In all cases, the linearity of the plots essentially extends through the origin (not shown). The slopes from these plots (60.4, 44.3 and 81.4 μV*sec/cpm for AP, 3-OH-AP and GS-AP, respectively; r > 0.99 for all three compounds) were used to calculate the relative amounts of the two metabolites in reactions with unlabeled AP as substrate in later experiments.
Other methods and materials.
Microsomes from combined liver or olfactory mucosa of 6 to 10 mice or OM S9 fractions from individual mice were prepared as described previously (Su et al., 1996;Gu et al., 1997). Tissues from C57BL/6J mice were obtained from a breeding stock maintained at the Wadsworth Center. Tissues fromCyp1a2(−/−) mice and wild-type littermates were obtained from breeding stocks maintained at the University of Cincinnati. Preparation and initial characterization of theCyp1a2(−/−) mice have been reported previously (Lianget al., 1996). Only males (∼20–25 g b.wt.) were used. For induction of CYP2E1, male C57BL/6J mice were given 1% acetone in tap water in place of drinking water for 7 days, whereas the control groups received water only. Protein concentrations were determined by the bicinchoninic acid method (Pierce, Rockford, IL) with bovine serum albumin as the standard. P450 concentrations in microsomes or purified preparations were determined according to the procedure of Omura and Sato (1964). Absolute spectra of purified CYP2A5 and CYP2G1 were recorded at room temperature using a Cary 3E spectrometer (Varian). Purification of NADPH-P450 reductase and b5from rabbit liver microsomes has been described in an earlier study (Ding and Coon, 1994). Purified rat CYP1A1 was a gift from Dr. Laurence Kaminsky of the Wadsworth Center. Metyrapone and unlabeled AP were from Sigma; 8-methoxsalen and 5-methoxsalen were from Aldrich. Other materials were obtained as described previously (Ding and Coon, 1988,1990; Peng et al., 1993; Ding et al., 1996).
Spectral and electrophoretic characterization of purified, heterologously expressed CYP2A5 and CYP2G1.
Bacteria-expressed CYP2A5 and SF9 cell-expressed CYP2G1 proteins were purified to electrophoretic homogeneity using conventional liquid-chromatographic methods after detergent solubilization of membrane preparations. As shown in figure 1A, a single band was detected by silver stain in purified preparations of CYP2A5 (lane 1) and CYP2G1 (lane 2). The two proteins migrated to about the same position and were detected as a single band when mixed (not shown). Efforts to resolve the two proteins by gel electrophoresis using different concentrations of acrylamide and by varying the length of the gels have so far been unsuccessful. For reasons not understood, mouse CYP2G1 was not stained as intensely as CYP2A5 by silver, although the two proteins were stained to a similar extent by Coomassie blue (data not given); different amounts of protein were applied in figure 1 to yield similar intensities. The specific contents of P450 in the purified preparations were ∼11.0 and ∼6.8 nmol/mg protein for CYP2A5 and CYP2G1, respectively, determined by CO-difference spectroscopy using an extinction coefficient of 0.091 μM−1 cm−1(Omura and Sato, 1964). The reduced carbonyl difference spectra exhibited maximal absorbance at 449 nm for both cytochromes with no peak at 420 nm (not shown) and absolute spectra of the cytochromes (fig. 1B) indicate that both are predominantly low spin in the ferric form, with maximal absorbance at ∼415 nm. The spin state of mouse CYP2G1 in the ferric form contrasts with that of rabbit CYP2G1, which is high spin (Ding and Coon, 1988).
Electrophoretic and spectral analysis of purified, heterologously expressed CYP2A5 and CYP2G1. A, Purified CYP2A5 (0.15 μg, lane 1) and CYP2G1 (0.5 μg, lane 2) were submitted to electrophoresis on a 7.5% SDS-polyacrylamide gel and detected by staining with silver. The positions of selected proteins in the prestained high-molecular-weight protein size marker (BioRad) are indicated. B, Absolute spectra of purified CYP2A5 and CYP2G1 were recorded at room temperature in 50 mM potassium phosphate buffer, pH 7.4, containing 20% glycerol and 0.1 mM EDTA.
Immunoblot analysis of CYP2A5 and CYP2G1 in mouse OM microsomes.
CYP2A5 and CYP2G1 can both be detected on immunoblots by antibodies to CYP2A10/11 or to rabbit CYP2G1 (Hua et al., 1997; Ding X, unpublished observations); highly specific antibodies that can distinguish the two isoforms are not currently available for a precise determination of the level of each isoforms in OM microsomes. A polyclonal rabbit antibody against purified, heterologously expressed CYP2A5 was obtained in this study. However, as shown in figure2, although this antibody (termed anti-CYP2A5) detected only a single band in OM microsomes (lane 1), it reacted with both CYP2A5 and CYP2G1 on immunoblots (lanes 2 and 3). Further quantitative analysis indicated that anti-CYP2A5 produces approximately equal band intensities with equal amounts of the two isoforms applied; thus, it can be used to estimate the combined level of the two proteins on immunoblots. In experiments not presented, a quantitative immunoblot analysis with purified CYP2A5 or CYP2G1 as a standard indicated that the combined level of the two isoforms in mouse OM microsomes is ≥150 pmol/mg protein, or >35% of the total microsomal P450 determined spectrally.
Immunoblot analysis of CYP2A5 and CYP2G1. Mouse olfactory microsomes (50 ng of protein, lane 1) or purified CYP2A5 (0.5 ng, lane 2) or CYP2G1 (0.5 ng, lane 3) were analyzed on immunoblots with anti-CYP2A5 (1 μg/ml).
Metabolic activation of DCBN.
The activities of purified CYP2A5 and CYP2G1 in metabolizing DCBN, an OM-specific toxicant, to DCBN-protein adducts were examined and compared with those of mouse liver and OM microsomes. As shown in table1, OM microsomes were ≥20 times more active (per mg of microsomal protein) than liver microsomes in the formation of DCBN-protein adducts, with 33 to 82 times higher turnover numbers (per nmol P450) at 30 and 3 μM DCBN, respectively. The activities of the purified, reconstituted, cytochromes were initially examined with BSA added as a donor of sulfhydryl groups. With BSA present, both CYP2A5 and CYP2G1 were active toward DCBN, with turnover numbers similar at 3 μM substrate and higher for CYP2A5 at 30 μM substrate but ≥20 times lower than that of OM microsomes. However, the addition of boiled mouse nasal or hepatic microsomes led to a substantial increase in the turnover numbers for both isoforms, whereas the addition of purified cytochrome b5 led to further increases in the activities of CYP2A5 and CYP2G1 to about one half (for CYP2A5) and one third (for CYP2G1) of that found in OM microsomes and much higher than activities seen in liver microsomes. Activity was not detected in control reactions containing boiled microsomes alone, with purified P450s omitted (not shown). These data indicate that both isoforms are active toward DCBN and that the dramatic difference in DCBN metabolic activities in liver and OM microsomes is not due to differences in the level of protein targets for adduct formation.
Metabolic activation of DCBN by CYP2A5, CYP2G1 and mouse liver and OM microsomes
Metabolic activation of AP.
Purified CYP2A5 and CYP2G1 were found to be active in metabolizing AP to 3-OH-AP and, in the presence of GSH, to GS-AP in a preliminary study (Genter et al., 1998). The activities of purified CYP2A5 and CYP2G1 in metabolizing AP to AP-protein adducts as well as 3-OH-AP and GS-AP were further examined here and compared with those of mouse liver and OM microsomes. The HPLC profiles of AP metabolites produced by CYP2A5 in a reconstituted system are shown in figure3. The only soluble metabolite detected in the absence of GSH with either purified P450s or in liver and OM microsomal reactions (not shown) was 3-OH-AP. With the addition of GSH, GS-AP was detected as the predominant product in addition to 3-OH-AP in all reactions. Three additional, minor, peaks, which may represent unidentified products, were also detected (fig. 3D, with retention times of 4.0, 6.0 and 11.5 min, respectively); the small peak at ∼11.5 min was also detected when 3H-GSH was used with unlabeled AP as substrate (data not shown), indicating that it may be a degradation product of GS-AP.
HPLC profiles of AP metabolites generated by CYP2A5 in a reconstituted system. Complete reaction mixtures contained 50 mM potassium phosphate buffer, pH 7.6, a reconstituted system containing 0.2 μM CYP2A5, 0.6 μM NADPH-P450 reductase, 30 μg/ml dilauroylphosphatidylcholine, 0.5 mM 14C-AP and 1 mM NADPH. Reactions were initiated by the addition of NADPH and were carried out at 37°C for 10 min (B and D). Control reactions (A and C) were terminated before the addition of NADPH. To determine GS-AP formation, GSH was added to the reaction mixtures at 10 mM (C and D). The metabolites were separated by reverse-phase HPLC and detected by UV absorbance (top: AU, absorbance unit) and radioactivity with an on-line radioactivity detector.
The turnover numbers of AP metabolism by purified P450s or microsomal preparations are shown in table 2. OM microsomes were 8 to 17 times more active (per mg of microsomal protein) than liver microsomes in the formation of the three products. The purified isoforms were also active in the formation of the two metabolites and AP-protein adducts, with turnover numbers higher than those found in liver microsomes. However, whereas the rates of formation of 3-OH-AP and GS-AP by purified CYP2A5 and CYP2G1 were close to those found in OM microsomes, the rates of protein adduct formation (per nmol P450) were 4 to 7 times lower in the reconstituted system, with or without (not shown) BSA present, than in OM microsomal reactions. In contrast to findings with DCBN-protein adduct formation, the addition of b5 or boiled liver or nasal microsomes did not change the rates of AP-protein adduct formation, andb5 had no effect on formation of 3-OH-AP and GS-AP (data not shown). In other experiments not presented, the effects of nondenatured microsomal fractions on AP-protein adduct formation were also examined. The addition of liver or nasal microsomes to reaction mixtures containing reconstituted CYP2A5 led to a small increase of overall activity, which, after deducting the activity seen in reactions with the microsomes alone, amounted to ∼2 to 3 times of the activity seen in reactions with reconstituted CYP2A5 alone but was still lower than nasal microsomal activities.
Metabolic activation of AP by CYP2A5, CYP2G1 and mouse liver and OM microsomes
Role of CYP2A5 and CYP2G1 in OM microsomal metabolism of DCBN and AP.
The role of CYP2A5 and CYP2G1 in microsomal metabolism of DCBN and AP was examined with the rabbit anti-CYP2A5 antibody used in immunoblot quantitation. In experiments not shown, anti-CYP2A5 IgG completely inhibited coumarin hydroxylase activity of both CYP2A5 and CYP2G1 in a reconstituted system when added at 5 mg IgG/nmol P450. Thus, the effects of various amounts of anti-CYP2A5 IgG on microsomal metabolism of AP to AP-protein adducts and of DCBN to DCBN-protein adducts were examined, with preimmune rabbit IgG added to maintain a constant concentration of IgG in all reactions. As shown in figure4, a concentration-dependent inhibition of OM microsomal activities was observed with both substrates, with maximal inhibitions of ≥80% of control activities achieved at 3 mg IgG/mg microsomal protein, confirming that one or both of these isoforms play a major role in AP and DCBN metabolism in OM microsomes. The addition of preimmune IgG alone did not cause any inhibition of the activities (not shown).
Immunoinhibition of OM microsomal AP and DCBN metabolism by anti-CYP2A5. Reaction mixtures contained 50 mM phosphate buffer, pH 7.4, 0.5 mM 14C-AP or 30 μM14C-DCBN, OM microsomal preparation (0.2 mg protein/ml for AP or 0.1 mg protein/ml for DCBN), various amounts of anti-CYP2A5 IgG or preimmune IgG added to maintain a constant level of total IgG in all reactions (0.6 mg/ml for AP or 0.3 mg/ml for DCBN) and 1 mM NADPH. The mixtures were incubated on ice for 15 min and then at 37°C for 3 min before addition of NAPDH. Reactions were carried out at 37°C for 10 (for AP) or 30 (for DCBN) min, and protein adducts were quantified as described in Materials and Methods. The activities are shown as a percentage of rates determined in control reactions to which preimmune IgG alone was added. The values shown represent the average of two determinations with differences of <20% of the mean. The anti-CYP2A5 antibody inhibits both CYP2A5 and CYP2G1.
The effects of two potential CYP2A5 inhibitors, 5-methoxsalen and 8-methoxsalen, on DCBN-protein adduct formation in reactions with CYP2A5 and CYP2G1 and with mouse OM microsomes were also examined. These compounds are known to inhibit the coumarin 7-hydroxylase activity in mouse liver microsomes (Maenpaa et al., 1993), and 8-methoxsalen has been found to inhibit heterologously expressed rat CYP2A3 (Liu et al., 1996). As shown in figure5, A and B, the compounds inhibited the activity of the two isoforms as well as that of OM microsomes to a similar extent, suggesting importance of one or both of these isoforms in metabolic activation of DCBN. However, 8-methoxsalen is a more potent inhibitor than 5-methoxsalen; with substrate present at 3 μM, a 50% inhibition was achieved with ∼2 μM 8-methoxsalen or ∼10 μM 5-methoxsalen. Similar results were obtained with metyrapone (fig.5C), a known inhibitor of DCBN toxicity in vivo (Brandtet al., 1990; Walters et al., 1993).
Inhibition of DCBN-protein adduct formation by 8-methoxsalen, 5-methoxsalen and metyrapone. The activities of CYP2A5, CYP2G1, and mouse OM microsomes in the formation of DCBN-protein adducts were determined with DCBN at 3 μM and 8-methoxsalen (A), 5-methoxsalen (B) or metyrapone (C) added at 0.5 to 200 μM. The other components of the reaction mixtures and the assay procedures were the same as described in the legend to table 1. The concentration of inhibitors is shown in logarithmic scale, and the activities are shown as percentages of control rates determined in the absence of an inhibitor. The values reported are the average of two experiments with differences of <20% of the mean.
The effects of 8-methoxsalen and 5-methoxsalen on OM microsomal metabolism of AP to 3-OH-AP (fig. 6, A and B) and AP-protein adducts (fig. 6C) were also examined and compared with the effects on the purified isoforms (fig. 6, A and B). As was found for DCBN metabolism, both compounds inhibited the activity of the two isoforms as well as that of OM microsomes to a similar extent, suggesting importance of one or both of these isoforms in metabolic activation of AP. In other experiments not presented, several other compounds, including flavone, α-naphthoflavone, 4-methylpyrazole, 1-aminobenzotriazole and coumarin, were found to inhibit the activities of CYP2A5 and CYP2G1 to similar degrees and thus could not be used to determine the relative roles of the two isoforms in OM microsomal metabolism.
Inhibition of AP metabolism by 5-methoxsalen and 8-methoxsalen. The inhibitory effects of 8-methoxsalen (A and C) and 5-methoxsalen (B and C) on the formation of 3-OH-AP in reactions with CYP2A5, CYP2G1 or OM microsomes (A and B) and on the formation of AP-protein adducts in OM microsomal reactions (C) were determined. AP was added at 0.5 mM, and the inhibitors were added at 0.5 to 200 μM. The other components of the reaction mixtures and the assay procedures were the same as described in the legend to table 2, except that no GSH was added when determining the formation of 3-OH-AP. The values reported are the average of two experiments with differences of <20% of the mean.
Role of CYP1A2 and CYP2E1 in OM microsomal DCBN and AP metabolism.
The possible role of CYP1A2 in OM microsomal metabolism of DCBN and AP were examined with Cyp1a2(−/−)mice and wild-type littermates. As shown in table3, rates of formation of AP- and DCBN-protein adducts were not decreased in OM S9 fractions fromCyp1a2(−/−) mice compared with Cyp1a2(+/+)mice, indicating that CYP1A2 does not play an important role in the metabolic activation of these compounds in the olfactory mucosa. For reasons not understood, S9 fractions from the CYP1A2-knockout mice had significantly higher activity toward DCBN than the wild-type littermates, although no difference was found with AP as a substrate.
CYP1A2 and CYP2E1 do not play a major role in the metabolic activation of AP and DCBN in mouse olfactory mucosa
The role of CYP2E1 was studied with mice treated with acetone to induce CYP2E1 level in the OM tissue. OM microsomes from acetone-treated mice had at least 3 times as much CYP2E1 protein as in microsomes from untreated mice, as demonstrated by immunoblot analysis with a monoclonal anti-2E1 antibody (data not shown). However, OM microsomal activities toward DCBN and AP were not different in the two groups (table 3). Thus, CYP2E1 does not play an important role in the metabolic activation of DCBN and AP in the olfactory mucosa. In other studies not presented, induction of CYP2E1 protein was not associated with an increase in OM microsomal activity toward 4-NP, a substrate frequently used for monitoring CYP2E1 activity in hepatic microsomes. Furthermore, both CYP2A5 and CYP2G1 were found to be highly active in 4-NP hydroxylation, with turnover numbers of 17.0 and 10.5 nmol/min/nmol P450, respectively, in a reconstituted system. Therefore, CYP2E1 may not play a major role in 4-NP metabolism and 4-NP hydroxylation may not be used for determining CYP2E1 induction in the olfactory mucosa.
Kinetics of AP and DCBN metabolism by purified CYP2A5 and CYP2G1.
To evaluate the relative importance of CYP2A5 and CYP2G1 in OM microsomal metabolism of AP and DCBN, we determined the kinetics of AP and DCBN metabolism in a reconstituted system. As shown in table4, CYP2A5 had only slightly lowerKm values than CYP2G1 did toward AP but much higher Vmax values in the formation of both 3-OH-AP and GS-AP. With DCBN, however, CYP2G1 had a much lowerKm and lowerVmax values than CYP2A5 did in the formation of GS-DCBN. Thus, it appears that CYP2A5 may play a greater role in OM microsomal metabolism of AP, whereas the relative roles of CYP2A5 and CYP2G1 in DCBN metabolism may be dose dependent, with CYP2G1 playing a more important role at relative low substrate concentrations.
Kinetic parameters of AP and DCBN metabolism by CYP2A5 and CYP2G1
Metabolic activation of DCBN is believed to go through 2,3- and 3,4-epoxy-DCBN, which form adducts with GSH or protein sulfhydryls (Ding et al., 1996). Several P450 isoforms are known to be active toward DCBN, including rat CYP2A3, rabbit CYP2A10/11, rabbit CYP2E1, human CYP2A6 and human CYP2E1 (Ding et al., 1996;Liu et al., 1996). AP is metabolized either directly or through yet unidentified intermediates to 3-OH-AP and N-acetyl-p-benzoquinone imine; the latter is believed to be the toxic intermediate that reacts with GSH or protein sulfhydryls to form covalent adducts (Hinson et al., 1980; Dahlin and Nelson, 1982; Harvison et al., 1988). Several P450s are known to catalyze this reaction, including human CYP1A2, CYP2E1 and CYP3A4 and their orthologs in other species (Morgan et al., 1983; Harvison et al., 1988; Patten et al., 1993;Lee et al., 1996; Kostrubsky et al., 1997; Zhouet al., 1997). However, definitive evidence for the P450 isoforms responsible for the metabolic activation of the two compounds in the olfactory mucosa has not been obtained in previous studies.
OM microsomes are much more active than hepatic microsomes in the metabolic activation of DCBN and AP, which may contribute to the tissue-selective toxicity of these compounds. Several lines of evidence were obtained in the present study that indicate that CYP2A5 and CYP2G1 play major roles in OM microsomal metabolic activation of DCBN and AP. First, purified CYP2A5 and CYP2G1 demonstrated activity toward the two toxicants in reconstituted systems. To this end, evidence was also obtained that CYP2A5 and/or CYP2G1 are abundant P450 isoforms in OM microsomes; the combined level of the two may account for >35% of total P450 in this tissue. Second, 5- or 8-methoxsalen provided parallel inhibition of AP or DCBN metabolic activation by the purified isoforms and OM microsomal preparations; similar results were obtained using an inhibitory anti-CYP2A5 antibody that also inhibits CYP2G1. In addition, the roles of two other P450 isoforms, CYP1A2 and CYP2E1, in the OM bioactivation of AP and DCBN were ruled out. There was no increase in the OM microsomal activities in acetone-treated mice compared with untreated mice, although OM microsomal CYP2E1 levels were 3-fold higher in acetone-treated mice. On the other hand, there was no decrease in the OM microsomal activities in Cyp1a2(−/−)mice compared with Cyp1a2(+/+) littermates. These results are consistent with a previous report suggesting that CYP2E1 may not play a major role in DCBN activation in rat OM microsomes (Eriksson and Brittebo, 1995) and with a recent study using Cyp1a2(−/−)mice indicating that CYP1A2 does not play a major role in the OM toxicity of AP (Genter et al., in press).
The effects of adding boiled liver or nasal microsomes on rates of DCBN- and AP-protein adduct formation in reactions with purified P450s are interesting. With DCBN, the addition of boiled microsomes instead of BSA as a donor of sulfhydryl groups led to a big increase in the rate of adduct formation. Similar increases were observed regardless of whether liver microsomes or nasal microsomes were used, indicating that the differences in liver and OM microsomal activities were not due to potential differences in microsomal proteins that can conjugate the reactive intermediates formed by P450 reaction. With AP, however, no increases were observed with addition of boiled microsomes. It remains to be determined whether the differential stimulation reflects different reactivity of the reactive intermediates formed from the two substrates.
The amino acid sequences of CYP2A5 and CYP2G1 are <60% identical (Huaet al., 1997). However, the two isoforms appear to be highly similar in immunological properties, substrate specificity and inhibitor selectivity. The two proteins are not resolved on immunoblots, and both react with several available antibodies prepared against CYP2A or CYP2G, including a monoclonal anti-CYP2A6 antibody from Gentest (data not shown). Both CYP2A5 and CYP2G1 are active toward a number of substrates, including coumarin, testosterone, progesterone, 4-NP, DCBN and AP, and both are inhibited similarly by all compounds tested to date. Furthermore, neither isoform appears to be inducible in the olfactory mucosa by chemical treatments. Although differences in kinetics parameters for DCBN and AP metabolism were found between purified CYP2A5 and CYP2G1, it is difficult at present to determine conclusively their individual roles in metabolic activation in OM microsomes. Attempts are being made to obtain isoform-specific antibodies to facilitate in vitro studies, and efforts to develop knockout mice models lacking either gene are warranted. The latter animal models will be very useful not only for resolving the role of the two P450 isoforms in microsomal metabolism but also for elucidating the role of these tissue-selective or -specific P450 forms in OM-specific toxicity of numerous foreign compounds, which is essential for reliable extrapolation of data from animal studies to risk assessment in humans.
We are grateful to Dr. Laurence Kaminsky for providing purified rat CYP1A1 and reading the manuscript. The authors gratefully acknowledge the use of Wadsworth Center Biochemistry and Tissue Culture core facilities.
Send reprint requests to: Dr. Xinxin Ding, Laboratory of Human Toxicology and Molecular Epidemiology, Division of Environmental Disease Prevention, Wadsworth Center, New York State Department of Health, Empire State Plaza, Box 509, Albany, NY 12201-0509. E-mail:xding{at}wadsworth.org
↵1 This study was supported in part by National Institutes of Health Grants ES07462 (X.D.), AG13837 (M.B.G.), ES06321 (D.W.N.) and P30-ES06090 (D.W.N.).
cytochromeb5
olfactory mucosal
Spodoptera frugiperta
DCBN
2,6-dichlorobenzonitrile
bovine serum albumin
immunoglobulin G
4-NP
p-nitrophenol
3-OH-AP
3-hydroxyacetaminophen
GS-AP
3-glutathion-S-ylacetaminophen
GS-DCBN
glutathione conjugate of DCBN epoxide
high performance liquid chromatography
Received November 25, 1997.
Accepted February 24, 1998.
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William Osler Abbott (1902 – 1943) was a United States physician, son of Dr. Alexander C. Abbott and Georgina Osler. His most notable contribution to the field of medicine was his part in the development of the Miller-Abbott tube, used in decompression and stenting of the small intestine, alongside Thomas Grier Miller, and also for devising the Abbot Rawson tube. Abbot received his MD in 1928 from the University of Pennsylvania. He died of myelogenous leukemia in Waquoit, Massachusetts on September 10, 1943.
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Pioneer BIOFOOD turns 30
Oct, 12/10/18 - 10:00
By Sara Önnebo
Section(s): Organic Entrepreneur
This year Swedish organic food producer, distributor, and wholesaler Biofood-Biolivs AB is celebrating 30 successful years in business. The company, which was founded by pioneer and trendsetter Denny Abrahamsson, continues to thrive in today's growing organic market. OWN talked to Marie Lavén, CEO at Biofood, about the company's journey, food trends, and organic sales in Sweden.
"Denny Abrahamsson has his roots in the environmental- and vegetarian movement. He started the organic food store Gryningen AB in Stockholm in the mid-70s because he wanted to eat organic food or "non-toxic food" as people used to call it in those days," explains Marie.
In the beginning, Denny drove all over Europe seeking organic and vegetarian food products for his store. Some of his early suppliers, for example, Horizon in the Netherlands, still exist today. Soy and mung beans, brown rice, seaweed and fermented food such as miso and tamari, part of the macrobiotic diet, were trending at the time.
When Marie joined the company in 1993 the organic food movement in Sweden was in its infancy. Although the business was growing, it still faced the challenge of educating the end customer about its products.
"I used to travel all over Sweden conducting demos at health food stores and informing customers about the meaning of organic food and healthier alternatives," remembers Marie. "Those first years I knocked on many doors and hardly any opened. But slowly more and more doors opened," says Marie who emphasizes that those experiences have been invaluable for Biofoods's development. "It takes time to make changes. You can't do it alone. Biofood was just a small part of the organic movement, but we all had a real sense of achievement and joy. The natural health food stores are the real heroes!" says Marie.
Eventually, their hard work started to pay off. Apart from distributing products from other companies, Biofood also has its brand to sell grains and pulses such as quinoa, amaranth, lentils, and rice. Denny set up the brand 30 years ago to supply consumers and the food service industry with a basic range of organic foods. Marie emphasized that Biofood's brand does not compete with their imported products, which consists of more than 2000 articles, including approximately 10 percent of skin care products.
Nut-based butter, with or without salt, crunchy or not crunchy, has been popular from the very start. Perhaps even more so today, thanks to the raw food and plant-protein trends where nut butter is added to smoothies or used in food balls. Demand for naturally fermented foods slowed down, but they are again receiving a lot of attention due to their remarkable health benefits. "And after some years of decreased interest for seaweed, there is now a shift towards searching for vegetarian alternatives in our oceans," says Marie.
Biofood also launched one of the first energy food bars in Sweden. Swedes were reluctant at first. Unlike the Norwegians, who already had a long tradition of bringing healthy snacks when going for hikes, the Swede's did not embrace the product immediately. Today, the energy food bar market in Sweden has exploded.
"We also introduced quinoa and whole spelt very early, to replace potatoes that were the staple food. Later these products became very popular because of diet-trends like the low-GI-diet and the wholemeal trend in the 90s," says Marie.
Not every new product introduction, however, has had a favorable outcome. The timing is also vital for success. "We are one of the trend-setters in Sweden when it comes to new alternatives, but sometimes we are a little too early!" says Marie. "We launched some healthy alternatives to soft drinks, but that didn't work out as well as we had hoped at the time."
Biofood welcomes solicitation from brands and companies looking for distribution in Sweden. "We are always looking for unique products that can fill a gap in the market. The product must be in line with our portfolio. It has to be sustainable, organic, fair-trade, free of plastic, healthy and natural in all senses of the word."
The company is cautious with the selection process to ensure that each product follows Biofood's values and is suitable for the Swedish market. "The Swedish authorities are tough, which can sometimes generate questions from foreign suppliers. We have to be able to prove to the Swedish authorities that we have control of the whole chain, from ingredients, production, labels, claims and so on," Marie explains.
The Swedish consumer associates organic food with better health, higher quality, and less environmental impact. However, many consider that products with organic certification are expensive, which poses a challenge. Sweden's best-known label for organic food and beverages is KRAV. Its standards fulfill the regulation of organic production at European and international levels and are in some cases even stricter. Besides organic farming methods and animal welfare, it also considers health, social responsibility, and climate impact.
Denny and Marie also attended exhibitions to stay in constant contact with the stores and consumers. Trade fairs offer an excellent way to get inspired, finding out about new products and meet with suppliers. Every year Marie visits BIOFACH, and the company exhibits at the Natural Products Scandinavia & Nordic Organic Food Fair in Malmo.
"I normally also try to visit the SANA trade show in Italy, also celebrating its 30th anniversary this year. It has become something of a tradition. The SIAL exhibition in Paris and Expo West in Anaheim are also on my wish list."
In 2001, Denny retired and sold Biofood to Marie and three of their colleagues who continued on his mission of an organic, healthy and vegetarian lifestyle to develop the company further. Marie has been the sole owner, after a joint decision, since 2010.
Although Biofood does business with retailers in other Scandinavian countries Sweden is its primary market. "We distribute to independent health food stores as well as established chains, either directly or via central warehouses for the stores that are members of a chain. We also deliver to large grocery chains, pharmacy-chains and independent, and to food service businesses."
Biofood with its 45 employees is indeed a company that "walks the talk." On top of supplying Sweden with sustainable products, they work according to an internal environmental manual to continuously improve on energy efficiency, transportation, and packaging. Moreover, the office, which is close to the original Gryningen organic food store where it all started, has been designed with great consideration for the environment and the wellbeing of the staff.
"We wanted to create a nice and welcoming atmosphere. The wooden desks and tables are handmade, and the walls are filled with pictures of forests, trees, and plants. Our office has also been Feng Shui certified by the Nordic School of Feng Shui," says Marie.
The organic sector is booming with almost three million producers worldwide. The United States continues to lead the market followed by Germany, France, and China. Denmark has the most pro-organic consumers in the world with more than 50% of the population buying organic food every week. Organic sales are also on the rise in Sweden and grew by almost 10% during 2017. Public procurement, backed by political decision at both national and local levels, for schools, hospitals and kindergartens have been a critical driving force of this development. Sweden has one of the highest organic per capita consumption in the world, only surpassed by Switzerland and Denmark.
The key to Biofood's success, in Marie's opinion, is high-quality products, and their commitment to providing top service and a strong relationship with customers built on trust and good communication. Biofood is a company that cares for its customers, suppliers and the environment. They will continue to implement this approach in the future. Another factor is the motivation to build a strong team spirit and camaraderie among the staff.
Biofood enjoys a relatively flat organization where titles are not what is essential.
"We believe very much in our values and hope that it is conveyed to our customers and suppliers. It's like a marriage; you have to prove yourself every day and be inspired about your work."
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Brazil fines US trading firms, local farmers over deforestation
Sikkim receives UN award for becoming world's first 100% organic state
Ginger Diamonds add spark to the candy aisle and more
Organic Food Iberia will celebrate Spain's leading role in the EU organic sector
A circular economy for the Amazon nut
Candela: 30 years preserving the forest
Fermentino: the art of fermenting nuts
Regenerative farming will lead to a sustainable supply chain for baru
UHTCO: A 10-year journey in Europe of excellence and quality
Protein content on packaging powers sales growth
Agave: the sweet revolution in French pastry making
Coconut palm sugar: untapped potential
Trend to natural, minimally processed sweeteners a boost for Sipal
Natural sweeteners rise with growing health concerns over refined sugars
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Fine Chocolate Today
Fairtrade steps up living income but cocoa price needs review
Major cocoa suppliers pledge action on deforestation
'Dirty' Cocoa' fells West Africa's rainforest
Pacari certified B Corp, flies first class with Emirates
UHTCO introduces new low glycemic chocolate
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Certification & Legislation
French mayors want glyphosate ban while Bayer adds organic pesticides
Is the organic concept & certification being diluted?
Consumers will place more scrutiny on plant-based foods
Amazon fires turn deadly
GMO crop deregulation threat to organic producers and exporters
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Chia World
Chia tops Google search data
Chia de France supports research
Official launch of "Chia de France"
First chia cultivated in France
Bolivian Chia Sector Proposes 'Clean Chia' Initiative
View all articles under Chia World