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SEAC Toggle navigation
LSE Southeast Asia Forum
ECR Network
Visiting Appointments
SEAC's friendly and welcoming staff provided me with a great environment in which to conduct research and progress with my thesis. I was able to consult LSE experts in the field, while the Centre's central London location and reputation enabled me to network with people I would not otherwise have had the opportunity to meet.
Daniel Peterson, Australian Catholic University
Prof Shin, Hyun Bang, Director
Hyun Bang Shin is Director of the Saw Swee Hock Southeast Asia Centre, and Professor of Geography and Urban Studies in the Department of Geography and Environment
Email: h.b.shin@lse.ac.uk
Dr Lee Mager, Centre Manager
Lee has overall operational responsibility for the Centre, including day-to-day administration, finances, communications, events and publications.
Email: a.l.mager@lse.ac.uk
Dr Yi Jin, Postdoctoral Research Assistant
Dr Jin is SEAC Postdoctoral Research Assistant for the Research Project "The Urban Spectre of Global China: Mechanisms, Consequences and Alternatives for Urban Futures".
Email: y.jin13@lse.ac.uk
Current SEAC Research Project Collaborators
Dr Sin Yee Koh
Dr Koh is Senior Lecturer in Global Studies, School of Arts and Social Sciences, Monash University Malaysia, and is Co-Investigator for the SEAC Research Project "The Urban Spectre of Global China: Mechanisms, Consequences and Alternatives for Urban Futures".
Dr Yimin Zhao
Dr Zhao is Assistant Professor in Urban Planning and Management, School of Public Administration and Policy, Renmin University of China, and is Co-Investigator for the SEAC Research Project "The Urban Spectre of Global China: Mechanisms, Consequences and Alternatives for Urban Futures".
The SEAC Steering Committee serves as a locus for consultation and deliberation, with the main responsibility to advise the SEAC Director on issues relevant to the Centre in line with its strategic objectives.
Steering Committee members in 2018/19
Professor Hyun Bang Shin
Centre Director, LSE SEAC and Professor in Geography and Urban Studies
Professor Tim Forsyth
Professor of Environment and International Development
Dr Nicholas Long
Assistant Professor in Anthropology
Dr Kirsten Schulze
Associate Professor in International History
Dr Lee Mager
Centre Manager, LSE SEAC
Centre Associates
SEAC Associates are academics at LSE or external researchers working on Southeast Asia who have an affiliation with the Centre. While SEAC Associates are invited to participate in and engage with activities at the Centre, they are not employed by SEAC and do not reflect the official position of the Centre.
Catherine Allerton, Associate Professor in Anthropology at LSE
Dr Catherine Allerton is a specialist in the anthropology of island Southeast Asia. She has conducted ethnographic research in the city of Kota Kinabalu, Sabah, with the children of Filipino and Indonesian migrants, and in rural Flores, Indonesia, with highland coffee farmers. Her work is broadly concerned with kinship, childhood, migration, place and landscape.
Dr Allerton's earlier research was conducted in the west of Flores, resulting in a number of articles, and a book, Potent Landscapes: Place and Mobility in Eastern Indonesia (2013). Her current research explores experiences of exclusion, belonging and potential statelessness amongst children in Sabah, East Malaysia. A number of forthcoming articles and a book will explore the complex politics of migration in Sabah, and the consequences for children born in the state to migrant and refugee parents.
Indonesia, Malaysia, Philippines
Sylvia Chant, Professor of Development Geography at LSE
Professor Sylvia Chant is Director of the MSc in Urbanisation and Development. She has conducted fieldwork relating to gender and development in Mexico, Costa Rica, the Philippines and Gambia, and has published widely on issues such as poverty, rural-urban migration, female employment and female-headed households.
Two books especially pertinent to the Philippines are: Women of a Lesser Cost: Female Labour, Foreign Exchange and Philippine Development (Pluto and Ateneo University Press, 1996), co-authored with Professor Cathy McIlwaine, and Gender, Generation and Poverty: Exploring the 'Feminisation of Poverty' in Africa, Asia and Latin America (Elgar, 2007). Her latest book, co-authored with Professor Cathy McIlwaine is Cities, Slums and Gender in the Global South: Towards a Feminised Urban Future (Routledge, 2016).
Tim Forsyth, Professor of Environment and Development at LSE
Professor Tim Forsyth has six years' professional experience within Southeast Asia as either a researcher or worker, and is fluent in Thai with skills in Bahasa Indonesia and Burmese. His research focuses on the politics of environmental policymaking, with a particular interest in understanding local environmental risk and livelihoods, and reflecting this knowledge in global environmental policies and assessments.
Professor Forsyth has worked on watershed and forest policies in Thailand and Indonesia; climate change policy and technology transfer in Southeast Asia; social movements and environmental governance; and the development of new, and inclusive, public-private partnerships as a basis for implementing global policy. He is the author of numerous books, articles and book chapters in journals concerning international development, environmental policy, and Southeast Asia.
Jürgen Haacke, Associate Professor in International Relations at LSE
Dr Jürgen Haacke teaches and researches on the international relations of Southeast Asia, especially the foreign and security policies of Southeast Asian states, the role and policies of major powers in relation to the ASEAN region, and regional multilateral cooperation. He has published particularly widely on ASEAN and Myanmar's foreign policy.
Dr Haacke is currently focusing primarily on a book manuscript on US Burma policy but is also pursuing research on the economic, political and security dimensions of UK-Southeast Asia relations in the context of 'Brexit'. He moreover continues to conduct research on foreign policy agency and practices in contemporary Myanmar.
From August 2016 until July 2018 Dr Haacke was Director of LSE SEAC. He is on sabbatical during the academic year 2018/2019.
Myanmar, ASEAN
Nicholas Long, Assistant Professor in Anthropology at LSE
Dr Nicholas Long is a specialist in the anthropology of Indonesia and the Malay World, with a particular focus on the ways political change influences the experience of self, agency, and relations. Professor Long's monograph, Being Malay in Indonesia, offers a new framework for the study of political decentralisation: one which foregrounds the affective and experiential dimensions of political change.
His fieldwork has also led him to develop fresh perspectives on many classic themes in the anthropology of Southeast Asia, including Malay identity, 'spirit beliefs', market cultures, poetry, memories of violence, and cross-border relations. He is currently working on two research projects, one investigating emergent forms of popular psychology in and their relationship to subjectivity, and one examining changing public perceptions of democracy.
Tan Sri Munir Majid, Chairman of Bank Muamalat Malaysia Berhad
Dr Munir Majid is currently Chairman of Bank Muamalat Malaysia Berhad, of CIMB ASEAN Research Institute and of the ASEAN Business Advisory Council, Malaysia. He is also the President of the ASEAN Business Club and a Visiting Senior Fellow at LSE IDEAS. Dr Majid's main interest is in the evolution of the region's contemporary international affairs under ASEAN. His focus on ASEAN, in turn, closely examines its economic integration process and extra-ASEAN economic engagements.
Dr Majid writes a fortnightly column in Malaysia's Star newspaper mainly on ASEAN matters. He has also written for Nikkei Asian Review and for the ISEAS-Yusof Ishak Institute of Southeast Asia Studies in Singapore, most recently contributing a chapter on private sector contribution in ASEAN economic integration in the Institute's Yearbook.
Malaysia, Geopolitics of Southeast Asia, ASEAN integration
James Putzel, Professor of Development Studies at LSE
Professor James Putzel is well-known for his research in the Philippines where he has maintained active research since 1984. His book, A Captive Land: the Politics of Agrarian Reform in the Philippines (1992), is recognised as a classic on the topic and remains influential in current policy actions and debates in the country. His research also includes work on nationalism, comparative politics of development in Southeast and East Asia, democratic transition, and the role of foreign aid and NGOs in development.
Professor Putzel served as Head of the International Development Department at LSE in 1999-2001 and was Director of the School's Crisis States Research Centre in 2001-2011. Aside from continuing work on land issues, his current research is focusing on politics and governance in crisis states including work on understanding 'state fragility', Political Islam in Southeast Asia and the rise of populist politics.
Danny Quah, Dean and Li Ka Shing Professor in Economics at the Lee Kuan Yew School of Public Policy, National University of Singapore
Prof Danny Quah's research interests include income inequality, economic growth, and international economic relations. His current research takes an economic approach to world order - with focus on global power shift and the rise of the east, and alternative models of global power relations.
Prof Quah is Commissioner on the Spence-Stiglitz Commission on Global Economic Transformation; Member, Executive Committee, International Economic Association; and Senior Fellow, Asian Bureau of Finance and Economics Research. He was also LSE SEAC's inaugural director in 2014-2016.
Geopolitics of Southeast Asia
Kirsten Schulze, Associate Professor in International History at LSE
Dr Kirsten Schulze works on communal and separatist conflicts as well as Islamism in Indonesia and the Middle East. Her publications on Indonesia include: The 'Ethnic' in Indonesia's Communal Conflicts: violence in Poso, Ambon and West Kalimantan (2017), The Islamic State and Southeast Asia (2016), and Transforming the Aceh Conflict: From Military Solutions to Political Agreement (2013). She is currently writing a book on the Ambon conflict titled Communal Violence, Mujahedin and Child Fighters: a history of the Ambon Conflict.
From 2014-2016 Dr Schulze was the Deputy Director of LSE SEAC, from 2012-14 she was the head of the LSE IDEAS Southeast Asia Program, and from 2004-2012 she ran the Indonesia seminar series at Chatham House.
Felicia Yap, LSE SEAC Associate
Dr Felicia Yap is an LSE SEAC Associate and an affiliate of the Centre of South Asian Studies in Cambridge. She was previously a Junior Research Fellow at Wolfson College Cambridge, a Scouloudi Research Fellow at the Institute of Historical Research in London and a Fellow in International History at LSE.
Dr Yap's research has centred on the effects of the Japanese occupation of Southeast and East Asia. Her interests include memories of conflict and captivity, the experiences of prisoners of the Japanese, the impact of the war on European and Asian women, as well as the evolution of Eurasian, Jewish and Portuguese communities in colonial Asia. She has begun a new comparative project on collaboration, resistance and espionage in wartime Macao.
Malaysia, Singapore, Borneo
Contact us Find out how to get in touch with LSE SEAC
Professor Saw Swee Hock Learn more about our founding donor


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Inside look at the new Mississippi Aquarium
15 years ago was one of the most detrimental days in not only this country's history but in Mississippi's history.
Hurricane Katrina virtually destroyed the entire Mississippi Gulf Coast. Taking the lives of over 200 people, destroying the homes of thousands, and causing billions of dollars in damage, the Category 5 hurricane is forever engrained in the minds of Mississippians.
However, today is a new day. This morning's opening of the Mississippi Aquarium in Gulfport epitomizes how far the coastal region has come since 2005.
"I remember coming down Highway 90 days after Hurricane Katrina and trying to find Gulfport…and nothing was left," former Governor Phil Bryant said. "We're sailing into the future. Literally, we sailed out of the worst natural disaster in America's history, and now, I think [the Gulf Coast] has become this iconic tourist destination."
Gulfport Mayor Billy Hewes shared similar remarks to the former governor.
"This is a monument to all of us, to our ability to recover, to the resiliency of our community and our spirit," he said at Friday's VIP ribbon cutting.
The state of the art, $93 million facility, which is directly across the street from the beach, includes everything from an aviary to both salt and freshwater habitats to plenty of room for events and social gatherings.
The Mississippi Aquarium will also prioritize animal research and conservation through its Aquatic Research Center (ARC). The ARC, through partnerships with Southern Miss and Mississippi State, will provide a hands-on, educational experience for visitors, plus access to some of the nation's top veterinarians.
In total, the project took three years to fully come into fruition, but now that the doors are finally open, the aquarium is estimated to bring in over $100 million annually, according to Anthony Wilson of Mississippi Power.
For an inside look at the Mississippi Aquarium, you can scroll through the photo gallery below.
The post Inside look at the new Mississippi Aquarium appeared first on News Mississippi.

redpajama_set_name: RedPajamaCommonCrawl


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THE GENTLEMAN BOXER
ENTRY SONG "CONQUEST OF PARADISE"
BY VANGELIS
After Henry Maske was asked not to use his original entry music, the "Carmina Burana" by Carl Orff, he started looking for something similar for the part of his fight night that was so important to him, the entry. The fact that at the end of the road "Conquest of Paradise", first used in October 1994 in Halle (Westphalia) in the Garry Weber Stadium, was used as a new companion to open fight night, inspired many spectators additionally. The subsequent rapid increase in sales success of the single "Conquest of Paradise", which had been on the market some time before, and the LP "Vangelis - 1492: Conquest of Paradise" was no different.
BY ANDREA BOCELLI & SARAH BRIGHTMAN
For Henry Maske the day had come to say goodbye after a long career. Musically, too, he wanted to say goodbye to the audience that had meanwhile been "spoiled" on RTL in all respects. The voice of the Italian singer Andrea Bocelli caught his eye during the summer of 1996. Thereupon he contacted Jürgen Otterstein, at that time managing director of Eastwest Records, publisher of Vangelis, and expressed his wish to have his farewell song created with this artist and their "conte partirò" as a basis. A musical dream couple was quickly found. Sarah Brightman and Andrea Bocelli sang a version "Time to Say Goodbye" specially composed for this last evening of the fight. On November 23, 1996, both stars climbed into the ring in Munich's Olympia Hall to get the more than 12,000 spectators in the hall and almost 18 million people in front of the television in the mood for the next match between Virgil Hill and Henry Maske.
"CURSED TO FIGHT"
THE NEW ENTRY MUSIC UND ITS HISTORY
The story of "Cursed To Fight" begins in tranquil southern Baden. Henry Maske is a guest at Europa-Park, Germany's largest amusement park. He is particularly fond of one of the attractions: The curse of Kassandra. Strictly speaking, it's a melody that he can't get out of his head. He decides if he ever got back in the ring it would be to this music. When his decision is made to fight Hill again, he contacts his friend and owner of Europa-Park, Roland Mack, and the production studio powerstation.
The melody from the curse of Kassandra is to be expanded, recomposed and arranged to create the entry music. Several variants of the piece emerged within 2 months until Henry Maske decided on the final version shortly before his fight. The Greek-spoken opening credits are about Kassandra, the seer who curses all those who are responsible for her downfall.
The title of the Henry Maske song developed from this story: "Cursed To Fight" - cursed to fight.
The new composition is a musical mirror of emotions, of hope and anxiety, a struggle for the right decision, which dissolves in the brilliant finale and leaves no more doubt:
I'm back - and the curse has turned into a blessing.
In addition, Henry Maske has released four of his own CDs under the title "power & glory". All CDs were released by eastwest Records Warner Music, Hamburg.
Henry Maske entry music 2007
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Home » Latest Grants and Resources for Peace and Conflict Resolution
Latest Grants and Resources for Peace and Conflict Resolution
View the latest grant funding and resources for Peace and Conflict Resolution for NGOs, companies, startups and individuals around the world.
Public Diplomacy Small Grants Program in Ghana
Deadline: 26-Feb-23 The U.S. Embassy Ghana Public Diplomacy Section of the U.S. Department of State announces its Public Diplomacy Small Grants Program. Purpose of Small Grants: to strengthen ties and build relationships between the United States and Ghana through activities that highlight … [Read more...] about Public Diplomacy Small Grants Program in Ghana
CCP Synergy 2023: Calling NGOs working in the areas of Sustainable Development, Climate Justice, and more…
Deadline: 15-Mar-23 The ifa (Institut für Auslandsbeziehungen) has announced the applications for its Cross Culture Programme (CCP) Synergy Connecting Civil Societies to promote tandem cooperation between civil society organizations from Germany and one of the 45 CCP partner countries that want … [Read more...] about CCP Synergy 2023: Calling NGOs working in the areas of Sustainable Development, Climate Justice, and more…
Peacecon 2023: New PeaceBuilding Architectures For Today and the Future
Deadline: 8-Feb-23 In partnership with the United States Institute of Peace (USIP), the Alliance for Peacebuilding (AfP) is proud to announce the theme for PeaceCon 2023: Beyond Fragile Ground: New Peacebuilding Architectures for Today and the Future, taking place in-person and virtually, May … [Read more...] about Peacecon 2023: New PeaceBuilding Architectures For Today and the Future
Call for Proposals: Fund for Local Cooperation 2023-2024 (Ethiopia)
Deadline: 30-Mar-23 The Embassy of Finland Addis Ababa is seeking proposals for its Fund for Local Cooperation (FLC) 2023-2024 to support local civil society organizations. The FLC complements bilateral development cooperation projects of Finland in Ethiopia. Goals The FLC is expected to … [Read more...] about Call for Proposals: Fund for Local Cooperation 2023-2024 (Ethiopia)
UN Voluntary Trust Fund for Victims of Trafficking in Persons launches 7th Call for Proposals
Deadline: 24-Feb-23 The UN Voluntary Trust Fund for Victims of Trafficking in Persons has launched its Sub-grant Programme 1 of the seventh Call for Proposals to provide essential humanitarian, legal and financial aid to victims of trafficking in persons through established channels of … [Read more...] about UN Voluntary Trust Fund for Victims of Trafficking in Persons launches 7th Call for Proposals
Nigeria: Youth Leadership Development Programme 2023
Deadline: 10-Feb-23 The Youth Leadership Development Programme is now open for applications to empower, equip, and support young change leaders who are Nigerian undergraduates under the age of 30, by providing them with the knowledge, abilities, and supportive environment needed for them to … [Read more...] about Nigeria: Youth Leadership Development Programme 2023
You Can Apply for Funding for Development Projects under the Polish Aid Program
Deadline: 28-Feb-23 The Embassy of the Republic of Poland in Dakar has launched the Competition for Co-financing Development Projects under the Polish Aid Program. Focus Areas Project applications submitted to the Polish Embassy in Dakar must relate to one of the following areas: Peace, … [Read more...] about You Can Apply for Funding for Development Projects under the Polish Aid Program
CdR/Lab Communication for Humanitarian Action in Colombia
Deadline: 1-Feb-23 Editorial Board (CdR) and the Forum of Humanitarian Organizations in Colombia open a call for the face-to-face workshop 'CdR/Lab Communication for humanitarian action in Colombia', which will be held in Bogotá on February 10 and 11, 2023. Purpose This training has with … [Read more...] about CdR/Lab Communication for Humanitarian Action in Colombia
U.S. Mission to Serbia: 2023 University Partnership Program
Deadline: 19-Apr-23 The Public Diplomacy Section (PDS) of the U.S. Embassy in Belgrade announces an open competition for organizations to submit applications for the University Partnership Program. Objectives The U.S. Embassy works to assist the Serbian people to strengthen democracy, … [Read more...] about U.S. Mission to Serbia: 2023 University Partnership Program
UNAOC Call for Applications: 6th edition of the Young Peacebuilders Programme
Deadline: 19-Feb-23 The United Nations Alliance of Civilizations (UNAOC) is launching the call for applications for the 2023 edition of its Young Peacebuilders programme, which for the second time will focus on Latin America and the Caribbean. The call is open to participants between 18 and 25 … [Read more...] about UNAOC Call for Applications: 6th edition of the Young Peacebuilders Programme
Grants for Improving Trade Controls through Port Management Practices
Deadline: 04-Mar-23 The Bureau of International Security-Nonproliferation is accepting applications for Improving Trade Controls through Port Management Practices. The purpose of this award is to address national security deficiencies in partner nations' export controls, public procurement, … [Read more...] about Grants for Improving Trade Controls through Port Management Practices
Grants for Advancing Strategic Trade Management in SE Asia
Deadline: 4-Mar-23 The Department of State's Bureau of International Security and Nonproliferation, Office of Export Control Cooperation (ISN/ECC) is pleased to announce a new funding opportunity through this Notice of Funding Opportunity (NOFO). Under this NOFO, ISN/ECC seeks proposals to … [Read more...] about Grants for Advancing Strategic Trade Management in SE Asia
Department of State's ISN/ECC: Counter IED Work in Tanzania – Mozambique Border
Deadline: 4-Mar-23 The Department of State's Bureau of International Security and Nonproliferation, Office of Export Control Cooperation (ISN/ECC) is pleased to announce a new funding opportunity through this Notice of Funding Opportunity (NOFO): Counter IED work in Tanzania - Mozambique … [Read more...] about Department of State's ISN/ECC: Counter IED Work in Tanzania – Mozambique Border
Call for Applications: N. AF Border Management Academy
Deadline: 4-Mar-23 The Bureau of International Security-Nonproliferation is seeking applications for its N. AF Border Management Academy. Under this NOFO, ISN/ECC seeks proposals to advance U.S. foreign policy and national security priorities by supporting initiatives that make decision-making … [Read more...] about Call for Applications: N. AF Border Management Academy
Kenya Persistent Ariel Surveillance Project – Apply Now!
Deadline: 4-Mar-23 The Kenya Persistent Ariel Surveillance Project is now open for applications to strengthen the capability of the Border Patrol Unit (BPU) in Kenya's Ministry of Interior to detect and identify transnational proliferation activities involving state and non-state actors at … [Read more...] about Kenya Persistent Ariel Surveillance Project – Apply Now!
NOFO: Advancing Strategic Trade Controls in Latin America
Deadline: 4-Mar-23 The Bureau of International Security-Nonproliferation has launched applications for Advancing Strategic Trade Controls in Latin America. The purpose of this NOFO is to provide legal-regulatory technical assistance, advice, and outreach to EXBS partners in the Western … [Read more...] about NOFO: Advancing Strategic Trade Controls in Latin America
Complementary Response Small Grants Program to Support Ukrainian Refugees (Lithuania)
Deadline: 4-Mar-23 The U.S. Embassy Vilnius of the U.S. Department of State announces an open competition for organizations to submit applications to carry out a program to support Ukrainian refugees in Lithuania. Objectives Applicants may submit a proposal focused on one or a few of the themes … [Read more...] about Complementary Response Small Grants Program to Support Ukrainian Refugees (Lithuania)
U.S. Embassy in Honduras – Public Diplomacy Small Grants Program
Deadline: 31-Mar-23 The U.S. Embassy Honduras Public Affairs Section (PAS) of the U.S. Department of State is pleased to announce an open competition for organizations and individuals to submit applications to carry out a program or project through its Public Diplomacy Small Grants … [Read more...] about U.S. Embassy in Honduras – Public Diplomacy Small Grants Program
Nominations Open for IIE Victor J. Goldberg Prize for Peace (US$20,000 to be shared among 2 winners)
Deadline: 23-Feb-23 Nominations are now open for the IIE Victor J. Goldberg Prize for Peace to recognize outstanding work being conducted jointly by two individuals, one Arab and one Israeli, working together to advance the cause of peace in the Middle East. The two individuals whose work is … [Read more...] about Nominations Open for IIE Victor J. Goldberg Prize for Peace (US$20,000 to be shared among 2 winners)
Fulbright Foreign Student Program (Philippines)
Deadline: 30-Apr-23 The Fulbright Philippines is now accepting applications for its Foreign Student Program. The Fulbright Foreign Student Program is a scholarship opportunity for Filipinos to study at the graduate level (master's or doctoral studies) or pursue non-degree doctoral dissertation … [Read more...] about Fulbright Foreign Student Program (Philippines)
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OBSESSED: Tap into Human Values and Truths to Build a Brand People Love
by Nasdaq Entrepreneurial Center
Tue, January 26, 2021
2:00 PM – 3:00 PM PST
How to Build a Brand People Love From Day One with Emily Heyward, Co-Founder & Chief Brand Officer of Red Antler.
We welcome back Emily Heyward, Co-Founder & Chief Brand Officer of Red Antler, for a virtual session to demonstrate why big and small brands have a hugely significant role to play in driving our culture, through times of great uncertainty and beyond.
During Emily's 12+ years building and running Red Antler, the leading brand company for startups and new ventures, she has not only witnessed the landscape evolve; she has helped shape it. Leveraging case studies from Red Antler's client roster and brands she admires, Emily will outline the modern brand-building principles inspired by the companies that get it right from the start. Learn how to position your brand in a singular way that connects to a human truth and provides runway for future growth and marketing opportunities.
In particular, we will address the following topics:
The best way to create connection with your consumer is to defy their expectations.
Innovation should always be in service of people, not "newness".
Any brand needs to tap into human values and truths we all share.
OUR SPECIAL GUEST
Emily Heyward
Co-founder & Chief Brand Office of Red Antler
Emily Heyward is the Co-founder and Chief Brand Officer at Red Antler, the leading brand company for startups and new ventures. Emily works closely with founders to develop purposeful, strategic visions for their startups and has led branding efforts for top companies such as Casper, Allbirds, Betterment, and Prose. After beginning her career in advertising, she started Red Antler along with her co-founders to directly apply consumer insights and brand positioning to launching and growing new and beloved businesses. In 2018, Red Antler made Fast Company's list of Most Innovative Companies in Marketing and Advertising and was dubbed the "cult brand whisperer".
Emily was named among the Most Important Entrepreneurs of the Decade by Inc. Magazine, and has also been recognized as a Top Female Founder by Inc. and one of Entrepreneur's Most Powerful Women of 2019. As a brand strategy expert, Emily is frequently interviewed by the press, with commentary appearing in Forbes, Bloomberg, Inc., Fast Company and Adweek. She's also been a featured speaker at TechCrunch Disrupt, 99U, and the Fast Company Innovation Festival. Her first book, Obsessed, will be published by Portfolio, a division of Penguin Randomhouse, on June 9, 2020.
Emily graduated Magna Cum laude from Harvard University, where she studied postmodern theory and consumer culture. Born and raised in New York, she currently lives in Carroll Gardens with her wife, son, and Brussels Griffon Sprout.
The Nasdaq Entrepreneurial Center is a 501c(3) public charity whose mission it is to make entrepreneurship accessible to all. We deliver world-class resources and mentoring to enable every entrepreneur across the globe to realize their maximum potential. Please join us and make your gift to ensure these transformative programs may continue to reach the underserved.
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Listen to an excerpt of DJ Ceremony & DJ Danny White's sets at "MOVEMENT." Recorded live at Passenger Bar in Williamsburg, Brooklyn on October 3, 2015.
With 2015's "Music Complete," New Order returned with their 1st album of newly recorded material since 2005's "Waiting for the Sirens' Call." To mark the occasion, & to pay tribute to one the most legendary Post-Punk / Dance bands of all time, DJ Ceremony & DJ Danny White hosted a release party for "Music Complete," and spun a night of exclusively Joy Division, New Order, UK Post-Punk, & '80s Dance at Williamsburg's Passenger Bar.
Note: Due to licensing rules in the US observed by Mixcloud, users cannot currently scrub or rewind backwards within a show, only forward.
Listen to an excerpt of DJ Ceremony's set at "Oscillate Wildly". Recorded live at The Gallery At LPR in NYC on May 30, 2015.
My flagship party, "Oscillate Wildly" is an ecstatic 6-hour dance party in celebration of the music of The Smiths & Morrissey, from the most celebrated classics, to fan favorites, to the deepest cuts. The party also features UK Postpunk, Britpop, & Manchester sounds.
Listen to DJ Ceremony's set at "Pre-93"—Alternative Dance Party (Produced by DJ Danny White). Recorded live at Cameo in Williamsburg, Brooklyn on March 26, 2015.
The concept of this monthly dance party is to pay homage to music that was once considered "Alternative" (and danceable) before 1993, with a focus on the late '80s/early '90s. DJs spin anything from Synth-Pop, to Industrial, to Alternative Rock.


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About The World Incorporated
Drink Water Straight from the Ground: Where to Find Iceland's Natural Mineral Springs
I'm thirsty. Thirsty for some natural Icelandic spring water straight from the earth. I want to taste those Icelandic mineral springs again. Filtered by nature. Melted from glaciers. Cold. Earthy. Maybe a little bit of a metallic taste. MMM.…
Souvenir Story: The Yule Lads of Iceland
Souvenir Story: The Yule Lads of Iceland Meet Bjúgnakrækir. That's Icelandic for "sausage swiper." And this is his mother's cat, Jolakotturinn (The Yuletide Cat), who enjoys lounging in the sun, sleeping all day, and eating children. Yum. Every December…
In Search of the Secret Ice Cave
In Search of the Secret Ice Cave Silence. We're standing on the edge of Fjallsárlón. It's a glacier lagoon adjacent to it's more famous counterpart, Jökulsárlón. It's different, however. The water is murky brown like chocolate milk. Too opaque to peer…
Food in Iceland You Need to Try
When it comes to food in Iceland, one may not expect a small island hovering below the Arctic Circle to hold a fork in culinary delight. But it does. Oh yes, the food in Iceland is quite delicious. In a county…
What to Wear in Iceland
It's difficult enough to figure out what to wear on your average Monday. But figuring out what to wear in Iceland is a a whole new challenge. Especially when you're trying to figure out what to wear in Iceland…
9 Days in Iceland: A 9 Day Iceland Itinerary Around Ring Road & Beyond
9 Days in Iceland: An Iceland Itinerary Around the Ring Road & Beyond Many people have told me, this is the best Iceland itinerary I've seen! This makes me glad, because planning a trip to Iceland and determining an…
Where to Stay in Iceland
Where to stay in Iceland? There are so many Iceland accommodations available for the traveler, depending on budget and type of trip. Budget travel in Iceland? Consider camping, hiring a camper van, or hosteling. Luxury travel in Iceland? Check…
The Cursed Rocks of Iceland
If you're here to find out if you can take rocks from Iceland or take sand from Iceland or if it is illegal to take rocks/sand from Iceland, please scroll down to the end of this post where I…
Glorious Gluttony: Fermented Shark (hakarl) of Iceland
The wind and rain inhibited our sightseeing plans for Snaefellsnes Peninsula. The visibility was so poor, we drove around the iconic Kirkjufell and could only see a faint outline of the monolithic mountain. Exploring new restaurants and trying local cuisine…
7 Reasons You NEED to Visit Iceland NOW
A tiny country hovering close to the Arctic Circle has captured the international travel spotlight: Iceland. And for good reason. Have you seen how many times Iceland is listed as one of the most mesmerizing places on the planet?…
Hi! I'm Amanda!
A career woman with a travel addiction. Fueled by weekends and vacation days. For a long time I didn't think I could balance a full-time job and travel the world. I'm doing both, and I'm sharing my secrets & stories so you can do the same. Michigan travel blogger based in Metro Detroit.
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WASHINGTON — New-car dealerships are no longer required to provide printed copies of the Relative Collision Insurance Cost Information brochure to car and truck buyers upon request. President Barack Obama signed legislation this week that repeals the outdated mandate and eliminates the $1,000 fine dealerships face for failing to comply with the 1970s-era law.
"This bill removes a regulatory burden from dealerships that helps reduce our cost of doing business," said Bill Underriner, chairman of the National Automobile Dealers Association (NADA) and a multi-franchise dealer from Billings, Mont.
In an NADA survey of its members, 96 percent of new-car dealers reported that none of their customers had ever asked to see the brochure in the 21 years dealers were required to stock it.
The federal government and the Insurance Institute for Highway Safety, which compiles the data, can still make the information available to the public online or by other means.
Underriner commended the leadership of House sponsors Reps. Gregg Harper (R-Miss.) and Bill Owens (D-N.Y.), as well as Sens. Amy Klobuchar (D-Minn.) and Roger Wicker (R-Miss.), who directed the bill through the Senate.


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Student Voices 2016 Option 4: Community Working Together
By Staff on April 1, 2016
Option 4: Share an example of a community working together to accomplish a greater goal. What happened and how was it made possible? What kind of roles did members play along the way?
Sophia Florence, Grade 6, Greene Street Friends School
My art is the pride flag, but I made it my own. I created this for my art submission because it represents when a big community came together and fought for what is right, for same‐sex marriage! This effort took a long time, but they did not just give up; they kept fighting. Finally others felt the same way, and they all came together and won! By coming together to fight for what was right, other people opened up to what was right, and the community just got bigger with lots of communities all over fighting for the same reason. It made a great impact on lots of people all over the world.
I also decided on this issue because it is important to me and my family. We have good friends who could not get married; this win didn't just have an impact on them but also on my family.
Sophie Adler, Grade 9, Westtown School
One of the times I have been deeply involved with a community working together was when I was in fifth grade at West Chester Friends School in West Chester, Pennsylvania. At the time, I was part of a girls' running club that met after school, and we decided that we wanted to do a service project. We chose to do our service for a nonprofit organization called Soles4Souls. This organization donates used shoes that are still in good condition to people in Third World countries. Many of these people do not have proper footwear. Our group watched a short video about the people that receive the donated shoes, and I remember wanting to help them immediately. There were only about eight girls in our running group, so we all split up with most girls working in pairs. Not surprisingly, I worked with my best friend, Ellie.
Each pair was given a huge cardboard box to collect the donated shoes. We had to decide where to set up our box. Some girls set up their box at the grocery store, others at the running store. At the time, Ellie and I were both part of West Chester Meeting, so we decided to put our box there. I remember Ellie and I standing up at the end of meeting for worship to make an announcement about our project. It seemed like everyone agreed that this was a good cause. We had about three weeks to collect shoes. We constantly reminded people to bring in shoes. As the weeks went by, Ellie and I saw people bringing anywhere from one pair of shoes to three bags full. At the end of the three weeks, we were surprised to see that our box was full. We were also proud of our accomplishment.
I'll never forget the Friday afternoon when we all brought in our boxes. First we dumped the shoes in one giant pile. Next we had to find pairs of shoes, tie them together with rubber bands, and put them in a different pile. This process took about an hour and a half. We had tons of fun while working. We took advantage of the many shoe options, trying on all of the heels we could find and attempting to walk across the gym. Finally all the shoes became a mountain, and we tallied all of the pairs. We collected 1,007 pairs of shoes, and the story was featured in the Daily Local News. This was a very memorable experience.
If the Taxes Aren't Lowered, the City Is Going to Stop!
Carolina De Avila Castilho, Grade 11, Westtown School
In July 2013, with the increase of taxes in São Paulo, Brazil, people from all over the city went out in the streets to fight for a greater goal. The protest was primarily organized in opposition to a significant price increase for bus tickets, which would cause a huge impact, especially for the lower classes. Later on, however, more people from all different classes started to join in the protest to represent their own causes. They fought against corruption scandals; they fought against the bad quality of public hospitals and the lack of investment in education; they fought against the large amount of money that was used for construction for the 2014 World Cup and not for supporting the starving children in the favelas (Brazilian slums). Soon enough, people from every state of Brazil were in the streets, crying out for a better place to live and fighting for their rights.
I was born and raised in Brazil, living there for 16 years before moving to Berwyn, Pennsylvania, to live with my aunt and uncle to attend Westtown School for a year.
In this drawing, I recreated a famous scene from the protest. On the board that the people are holding there is a warning: "If the taxes aren't lowered, the city is going to stop!" This is an example of how a community can work together to achieve a greater goal. After that month of protests, corrupt politicians were arrested, taxes were lowered, and changes were made in our hospitals and schools. Now our leaders recognize that the people have a voice, and that it is louder than ever before.
Time to Step Up
Franklin Grear, Grade 9, Westtown School
Over the summer, I had the opportunity to travel to Helsinki, Finland, to become part of a new community. The trip was run by a nonprofit organization called Children's International Summer Villages (CISV), which works toward world peace. I participated in a Step Up camp called Say What!?. Little did I know going into it that this community would forever change my life.
There were nine countries represented at the camp: Japan sent three girls; Finland sent two boys and a girl; Italy, Hungary, China, the United States, Lebanon, Austria, and Poland each sent two boys and two girls. These teenagers were considered the participants of the camp. Each country also sent one adult leader. There were also four staff members from Finland and one from Mexico. Those are all of the members that made up the community.
Say What!? Step Up 2015, along with all other CISV Step Up camps, had a few goals, the main ones being to bring teenagers together to build global friendships, to take a little step toward world peace, and to help us all develop some leadership skills. From the day I got to the camp, I could tell that the next three weeks would be the best of my life. Each day I became closer with every person in the camp. You may be asking, "How did that happen?" One of the main reasons we were able to build such a strong community in three short weeks is that we recognized we were all different from one another. Those differences were not only recognized but cherished. The time I spent at this camp was the first time I went three weeks without hearing anything negative about somebody's race, religion, beliefs, political views, culture, country, or skin color. We all spoke English—some people didn't know as much as others, but we didn't let that stop us. The language barrier rarely formed even the slightest of problems. At meals nobody rushed to sit next to specific people because we all loved everybody there.
I think the thing that could make every community stronger is cherishing each other's differences like we did. When I was younger, I would always hear about people being accused of crimes due to their race, religion, or cultural heritage. Growing up this way, I never thought I'd see a place where race, culture, and religion existed only through positive perspectives. The absence of intolerance at Say What!? Step Up is one of the things that made it such a strong community. Another thing was simply that we spent time together. Participants spent time with staff, leaders with participants, staff with leaders. It all resulted in a formula that made, in my eyes, the ideal community. I never thought that I would see a perfectly peaceful place in which the only pain was saying goodbye. Will you be able to step up and make your community a better place?
Student Voices 2016
Option 1: What are the important elements of building a loving, safe, and supportive community?
Option 2: What are some ways to support and care for each member of your community?
Option 3: What happens when there are differences or conflicts within a community?
Option 4: Share an example of a community working together to accomplish a greater goal.
Thanks: Thank you to all of the participants of the third annual Student Voices Project!
Posted in: Spiritual Nurture / Quaker Training, Student Voices Project, Uncategorized, Unfeatured
art, Brazil, Carolina De Avila Castilho, Children, China, community, education, Finland, girl, language, process, store, United States, Westtown School, work
Student Voices 2016: Thanks
Student Voices 2016 Option 3: Conflicts in Community


2 Lads Sparkling Pinot Grigio is an ideal holiday wine that pairs well with a range of flavors, complementing everything from your favorite appetizers to entrees, even dessert! Plus, this wine brings an added elegance to any holiday table.
This time of the year is the right time for Pinot Noir, and Black Star Farms suggests their 2011 Arcturos Pinot Noir. The lighter body of the Pinot Noir is the perfect match for lighter meats and holiday fare.
Bowers Harbor Vineyard 100% Pinot Noir Rosé is another fantastic option this holiday season. Bursting with fresh watermelon, strawberry, and cranberry, this Rosé complements turkey and holiday traditions. Though it is considered a dry wine, it drinks with a variety of palettes, so enjoy this fruity and light spice.
This holiday season, reach for a Pinot Noir or a Dry Riesling – both complement earthly fall flavors and holiday spice. Brys Estate Vineyard & Winery Pinot Noir pairs beautifully with roasted vegetables, while their Dry Riesling will complement stuffing with apples and spice. Also, try the Dry Riesling with a rich butternut squash bisque or curried carrot soup. It's sure to please.
Chateau Chantal offers their Naughty & Nice Red and White wines during the holidays. Perfect as a fun wine gift for the season. Ranging from dry to sweet, there is a selection for everyone at the table among these four wines. The Naughty Red and White are dry, while the Nice Red and White are the sweet versions of the same wine blend.
The 40th Anniversary wine, the 2013 Etcetera Rosé, from Chateau Grand Traverse is the perfect pairing for a holiday feast. Its robust, off-dry flavors offer pepper-like qualities that are sure to hold up to any poultry or white meat dish. The creamy finish and off-dry style make it versatile to share with grilled vegetables or pasta dishes. This rosé wine is celebratory in nature, as it was created to embrace the 40 years Chateau Grand Traverse has spent crafting wines on Old Mission Peninsula.
The perfect pairing for a holiday prime rib with a creamy horseradish sauce or an herbed rack of lamb with rosemary and thyme is Peninsula Cellars award winning Cabernet Franc. This dry red wine, voted best-of-class dry red and double gold for two consecutive years, boasts rich mocha and deep berry flavors.
This holiday season, taste, learn, share, and enjoy wines from Old Mission Peninsula! The Wineries of Old Mission Peninsula's annual Day of Giving is the perfect time to stock up on these award-winning wines. On Saturday, December 13, the wineries will donate 15% of tasting room and online sales to Northern Michigan nonprofit organizations. Make a weekend out of it by pairing with the December 11 Ladies' Night shopping and noshing extravaganza in Downtown Traverse City!
Jutting north between the azure arms of Lake Michigan's Grand Traverse Bay lies Old Mission Peninsula. Situated at the globe's 45th parallel – an ideal climate for growing varietal wine grapes – the eight distinct wineries that comprise the Wineries of Old Mission Peninsula® and this stunning appellation have been well recognized by wine enthusiasts around the world.


The play takes place in the remarkable city of Vilna, a highly evolved center of civilization where Jews lived since the tenth century. In the 18th century, Vilna was the center of Jewish learning in Europe. Napoleon called it the "Jerusalem of the North". In 1916, of 148,000 inhabitants, 61,000 were Jews (over 41% of the population!) Never before has there been any place in the diaspora world with this Jewish demographic density.
"Vilna" is a play about the real people who sustained the civilized fabric of the Vilna Jewish community during its degradation in the Interwar years and its complete destruction by the Nazis in WWII. The play follows the lives of Motke Zeidel and Yudi Hirschmann as they grow up together and become central characters in the social and political events of their time.Their encounters with historical people and situations tell a story of heroic perseverance under extraordinary circumstances.
*Discounted pre-sale will be available until February 7th, 2019. Discounts available for universities and students. Contact email below for more information.
For more information, please contact irahfuchs@gmail.com.


The Achievement School District is looking to Westwood Elementary to help her find answers to one of the state district's longtime issues: lack of students.
That would bring the number of charter schools in Tennessee's largest district to 63 — far and away the most in the state.
The teachers, mostly black men, have turned it into a networking opportunity as well as a way to let off steam.
The troubles that at least two school districts had connecting to Tennessee's online testing system were not related to a slashed cable, the internet provider says.
Student anxiety, last-minute schedule changes, and hour-long holds with customer service hotline were common.
Ten charter organizations applied to Shelby County Schools ahead of the April 2 deadline to open 18 schools in 2019.
Eleven elementary and middle schools from four different charter organizations in Memphis have adopted the KIPP Wheatley curriculum.
The school board's vote comes just months after the district overhauled its charter school office to strengthen oversight of the growing sector.
In Freedom Preparatory Academy's inaugural graduating class, all 50 seniors are headed to college.


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remembr
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Put your brain in this tool and make it scale!

redpajama_set_name: RedPajamaGithub


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Intrado, TSG Global and WMC Global Form Strategic Initiative
Telecom Leaders Pioneer Blockchain Identity Solution to Address Major Telecom Issues
BOSTON--(BUSINESS WIRE)--Intrado, TSG Global and WMC Global have formed a strategic initiative to address key identity, privacy and security issues in enterprise communications using blockchain or distributed ledger technology (DLT).
The goal of the initiative is to enable enterprises to validate the information associated with their telephone numbers and prevent scammers and spammers from hacking their identity. The three telecom leaders will use the newly developed patented blockchain identity solution TNID (Telephone Number ID) to securely store, share and attest business information linked to a telephone number.
Enterprises and business process outsourcers can use TNID's "source of truth" record to manage every one of their Toll-Free and local phone numbers, thus enabling them to more easily deliver trusted consensual communications to consumers who have opted in.
By standardizing the identity process, TNID's private blockchain network will also dramatically reduce telecom fraud and robocalls, which was the main goal of the industry-wide STIR/SHAKEN initiative.
"We are excited to be one of a group of like-minded innovators dedicated to protecting sensitive customer data while enabling enterprises to deliver trusted communications," said Noah Rafalko, CEO of TSG Global. "With the global reach of Intrado together with WMC Global's monitoring and security, we are confident we can solve the issues that STIR/SHAKEN only begins to address."
About Intrado Corporation
Intrado Corporation is an innovative, cloud-based, global technology partner to clients around the world. Our solutions connect people and organizations at the right time and in the right ways, making those mission-critical connections more relevant, engaging, and actionable - turning Information to Insight. Intrado has sales and/or operations in the United States, Canada, Europe, the Middle East, Asia Pacific, Latin America, and South America. Intrado is controlled by affiliates of certain funds managed by Apollo Global Management, Inc. (NYSE: APO). For more information, visit www.intrado.com.
About TSG Global
TSG Global brings decades of experience in educating, designing, migrating and deploying the latest vetted communications technologies necessary for today's successful enterprises. With a laser focus on clients' needs, TSG Global unbundles the complex array of communications services and reassemble them in a portfolio that's custom built for each and every client based on their unique needs. For more information, visit www.tsgglobal.com.
About WMC Global
A 15-year leader in mobile threat intelligence, WMC Global delivers innovative solutions that promote positive mobile customer experiences, safeguard the messaging ecosystem, and protect the consumer digital lifestyle. Notably, WMC Global provides expertise in text message monitoring and compliance, and offers RISQ Score, a due diligence solution that evaluates an entity's risk profile based on historical organizational performance. Expanding beyond mobile and messaging to address all digital service sectors, WMC Global focuses on phishing and provides reporting, monitoring, mapping, and takedown of deceptive, fraudulent, and illegal global activity. To learn more about WMC Global, visit. www.wmcglobal.com.
Maria Verven
TSG Global, Inc.
Maria.Verven@TSGGlobal.com


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În chimia organică și în biochimie, un substituent este un atom sau o grupare de atomi care ia locul sau "substituie" un atom de hidrogen din catena principală a unei hidrocarburi. Termenii substituent, catenă laterală, grupă sau ramificație sunt folosite pentru a descrie ramificațiile unei catene principale,  deși sunt făcute unele distincții în contextul chimiei polimerilor.

Sufixul il este folosit la numirea compușilor organici care conțin un substituent cu legătură simplă, iar ilen / iliden și ilin / ilidin pentru cei cu legătură dublă și respectiv triplă.

Substituenții metanului
Un atom de carbon dintr-o moleculă, considerat a fi un substituent, poate avea următoarele denumiri, în funcție de numărul legăturilor de hidrogen legați de acesta și de tipul legăturilor formate:

Vezi și
 Grupă funcțională
 Catenă laterală

Referințe

Chimie organică

redpajama_set_name: RedPajamaWikipedia


Pretty much every restaurant has a kids menu. A lot of times the options for kids are pretty limited or perhaps not that healthy. Some places not only give kids more options but also better options. If you have kids, try these places where your kids are sure to find some menu item they enjoy, which makes for a better meal for everyone involved.
Sessions is a great place to bring the whole family. Here you'll find some really good things on the kids' menu. One of the items is a grilled cheese sandwich made with sharp cheddar. Another option is a turkey sandwich complete with cheese, lettuce, and tomatoes. This is a big sandwich, so if you have smaller kids, this can definitely be shared. The sandwiches are served with either fries or apple slices. Finally is one of those "secret menu" items: chicken and donuts, which is a twist on chicken and waffles. Chances are good that you won't have to box any of this if you order it for your kids.
This is the great place to bring the kids for a couple reasons. First, a variety of restaurants call 4th St. Market home. This is great news if you have kids who can never agree on where they'd like to eat. Each of your kids can choose to eat at a different place, and then you can gather as a family to eat on the patio or in any of the common areas. Second, each place has a kids' menu that goes beyond the usual burger and grilled cheese.
One issue with kids menus is that the frequently have very few options. Rutabegorz realizes that kids don't always want the same three things, so they provide options for their young eaters. Pasta is usually a safe bet with kids, and here they can order it with red or white sauce, or with butter and cheese. Macaroni and cheese is also an option as well as a cheese sandwich, a pizza bagel, and a quesadilla. Your kids are likely to find something they like on the menu here.
Technically Burntzilla doesn't have a kids menu. However, this place specializes in food that is just the right size for kids. Here you'll find sliders ranging from burgers to fried cheese. You'll also find Zillas, which are miniature versions of hot dogs that are perfect for small hands. Your kids are sure to find something they like at this restaurant where the food looks like it was designed for kids.
At most restaurants, you get the feeling that they offer a kids menu simply because they are expected to do so. The options for kids are usually few. Reunion Kitchen gives kids more options than just a burger or grilled cheese, although both of those are part of the kids' menu. Here kids can choose from items like fish and chips, pasta, or even a chicken teriyaki bowl. This place definitely goes beyond the standard kids' menu.


Tea tree oil is very effective for dandruff as it possess antibacterial and antifungal properties, with moisturizing benefits. Learn how to use it to treat dandruff!
The white patches of dead skin cells known as dandruff are telltale signs that your head is in a very bad condition, and having those white flakes scattered in your head and shoulders can be very annoying and embarrassing. In addition to the irksome itchiness it causes, dandruff can definitely ruin your appearance.
To remedy dandruff, we often subscribe to chemically loaded hair products that we see in advertisements, promising to manage the condition. Yet, come to think of it, some manufacturers of dandruff shampoos are actually not in the business of clearing dandruff down to its roots, or what use would be their business?
Essential oils on the other hand, have been utilized since antiquities to treat dandruff down to its core. Research shows that essential oils contain natural chemical components that effectively kill various pathogens such as bacteria, virus, fungus, and many more. One essential oil that promise to solution itchy dandruff problems is tea tree oil, an essential oil derived from Melaleuca alternifolia tree of Australian origin.
What makes tea tree oil very effective for dandruff is that it contains terpenes, a naturally occurring chemical that is proven to not only possess antibacterial and antifungal effects but also with fine moisturizing benefits. In addition, terpenes penetrate deep down into the cells to cleanse off any dead cells and quirky things that are making the scalp itchy.
Tea Tree Oil for Dandruff?
Mix thoroughly the carrier oil of your choice with the essential oil.
Massage hair and scalp with the mixture.
Leave the oil onto your scalp overnight or at least half an hour.
Wash your hair off with shampoo (natural product if possible).
Initially, do the procedure 2-3 times a week. Once dryness and dandruff is gone, decrease application once in a week.
Dandruff responds well to vinegar rinse, yet adding up tea tree essential oil to this classic rinse can amp up its effects. Follow the steps below to make your very own hair rinse.
Mix apple cider and tea tree oil, and combine them with spring water. Using about a teaspoon each time. Massage into the scalp every night before going to bed.


The National Monument of Solidarity commemorates Luxembourg soldiers who lost their lives in World War II and serves as a reminder to the future generations about their forefathers' suffering in the war against Nazi occupiers.
The country's entire population took part in the campaign to raise funds for the construction of the monument, which was designed by stained glass artist Francois Gillen and architect René Mailliet. The unveiling ceremony of the monument on October 10, 1971 was attended by Luxembourg officials and the general public, including members of the Resistance Movement.
Built in an architectural style that was fashionable at the time, the monument constitutes a stone chapel with walls of stained orange, violet and blue glass.


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Q: Chart Webpart: Show only integers? I have a chart web part using columns to display numeric values on the Y-axis and text values on the X-axis.
How do I set the chart to only show integers on the Y-axis? Right now it shows float numbers like the linked picture displays:

A: See the documentation about chart web parts here: http://office.microsoft.com/en-us/sharepoint-server-help/create-a-chart-by-using-the-chart-web-part-HA101889211.aspx
Pay special attention to the section about "Axes and Gridlines", specifically:
"In the Interval box for the major grid lines, type a number that corresponds to the interval that you want to use for the major grid lines. For example, if the chart displays data in units of 100, you might type 100 for the interval to display major grid lines."

redpajama_set_name: RedPajamaStackExchange


Evil demons are sieging the Heaven's gates! You are a cupid armed with a big bow and your aim is to prevent them from doing it. You have received a God's order: kill everyone who hasn't got a halo over his head. Have a good hunt!


Gaian Times eco-Magazine: Informative & thought-provoking articles, columns, poetry, puzzles & more! Regular Features include: Astrology, Esoterica, Poetry, Eco-Art & Photo Journals, info on Environmental Groups and more. Gaian Times has Charitable Objectives, that are legally stated in our Articles of Association.
Subscribers PLEASE BOOKMARK this page, for easy return! .


Disclosure: I was sent a DVD from the vendor in order to write up an honest review. All views shared are mine and mine alone.
"Join Mily, a funny and inquisitive 8-year-old girl, as she embarks on adventures hoping to answer the many questions she has about life, people, and the world. In this collection of twenty four episodes, join Mily and her friends as they investigate everyday situations from learning how to deal with fear to playing by the rules to using your imagination and so much more.
Whenever a particular situation raises a question worthy of examination, Mily is immediately on the case! From one episode to the next, through everyday situations, Mily introduces young viewers to the art of questioning in order to further develop an inquisitive mind."
My girls really enjoyed this DVD, which has a run time of 175 minutes. Having not seen this series before, they were excited to meet Mily, who is a funny and inquisitive 8-year old, who embarks on fun adventures while answering many questions she has about life, people and the world. She is just like my girls...full of questions and always asking away, hoping to be enlightened, or working towards finding answers for themselves.
To learn more about Mily Miss Questions and other wonderful educational titles like this one, head on over to NCircle today. You can find all DVDs offered by NCircle on their website and at retailers near you.


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TEXAS - CONTINUING MEDICAL EDUCATION (CME)
State laws and regulations that affect your medical practice
1. How many hours of Continuing Medical Education are required each year?
Physicians are required to complete 24 hours of continuing medical education every 12 months. At least 12 hours every 12 months, including 1 hour of ethics and/or professional responsibility education, must be completed through participation in formal CME activities, but all 24 hours may be completed in this category. This includes conferences, seminars, lecture presentations, grand rounds, case conferences, self-study courses, etc. which are formally designated for credit as: Category 1 of the Physician's Recognition Award of the American Medical Association; prescribed credit of the American Academy of Family Physicians; or Category 1A and 2B credit of the American Osteopathic Association.
Physicians may complete up to 12 hours of the required 24 hours by participating in informal CME activities such as conferences, seminars, grand rounds, self-instructional materials, reading clinically relevant medical journals, participation in patient care review activities, research/preparation time for medical presentations, up to 10 hours for publication of a medical article and up to 6 hours for volunteer services at a site serving medically underserved populations.
Yes. Excess formal or informal hours earned in one 12-month period may be applied toward the next year's requirements. A maximum of 48 excess credit hours may be carried forward and these hours must be applied within two years following the date of the registration period during which they were earned. A licensee under a two-year license period may apply up to 24 CME hours retroactively to the preceding year's annual requirement. These hours may be counted only toward one registration period.
3. How will the Texas Medical Board know whether a physician has completed the necessary hours of CME?
Documentation of attendance at formal CME activities can be obtained from the provider of the activity in the form of a CME reporting form, attendance, certificate, CME transcript or letter of verification of attendance.
Documentation to verify attendance should not be submitted to the Texas Medical Board with the application for registration. Documentation, however, should be retained by the physician for reference in reporting hours completed and in the event that verification is requested by the board in a random audit of compliance.
4. Are there any exemptions for CME?
Exemptions are subject to the approval of the executive director of the Texas Medical Board and must be requested in writing at least 30 days prior to the expiration date of the license. An exemption may be requested for the following reasons:
5. What is the penalty for not completing CME requirements?
Failure to obtain and report 24 hours of CME every 12 months at the time of license renewal will result in nonrenewal of the license until the physician obtains and reports the required CME hours. However, the executive director of the board may issue a temporary license for a period of up to 90 days to the physician who has not complied with the CME requirement. Note that this 90 day extension through the temporary license is at the discretion of the executive director and is not an automatic grace period. The temporary license not only allows the board time to verify the accuracy of information related to the physician's CME hours, but also allows the physician an opportunity to correct any deficiency so as not to terminate ongoing patient care. Any CME hours obtained after a physician's renewal date for the purpose of satisfying the CME requirement for the preceding year's licensure renewal, if above and beyond the 24 hours necessary for renewal, may be used to meet the CME requirement for the current year.
Failure to comply with the CME requirement for renewal of a license invokes the monetary penalty for late registration and may invoke Administrative Penalties as determined by the Disciplinary Process Review Committee of the board. A false report or statement to the board regarding CME hours is basis for disciplinary action by the board.
Texas Medical Association, Summary of Texas Medical Board Rules for Reporting CME Activities for Registration of the Medical License
Copyright Kern Augustine Conroy and Schoppmann, P.C. Used with permission.


Acetobacter sp. CCTCC M209061 could catalyze carbonyl compounds to chiral alcohols following anti-Prelog rule with excellent enantioselectivity. Therefore, the enzymatic characterization of carbonyl reductase (CR) from Acetobacter sp. CCTCC M209061 needs to be investigated.
A CR from Acetobacter sp. CCTCC M209061 (AcCR) was cloned and expressed in E. coli. AcCR was purified and characterized, finding that AcCR as a dual coenzyme-dependent short-chain dehydrogenase/reductase (SDR) was more preferred to NADH for biocatalytic reactions. The AcCR was activated and stable when the temperature was under 35 °C and the pH range was from 6.0 to 8.0 for the reduction of 4′-chloroacetophenone with NADH as coenzyme, and the optimal temperature and pH were 45 °C and 8.5, respectively, for the oxidation reaction of isopropanol with NAD+. The enzyme showed moderate thermostability with half-lives of 25.75 h at 35 °C and 13.93 h at 45 °C, respectively. Moreover, the AcCR has broad substrate specificity to a range of ketones and ketoesters, and could catalyze to produce chiral alcohol with e.e. >99% for the majority of tested substrates following the anti-Prelog rule.
The recombinant AcCR exhibited excellent enantioselectivity, broad substrate spectrum, and highly stereoselective anti-Prelog reduction of prochiral ketones. These results suggest that AcCR is a powerful catalyst for the production of anti-Prelog alcohols.
Chiral alcohols are vitally important building blocks for the synthesis of pharmaceuticals, agricultural chemicals, flavors, and special materials. Currently, both chemical and biological methods can be used to synthesize chiral alcohols. The most prospective biocatalytic method is the asymmetric transfer of hydrogenation to carbonyl groups catalyzed by reductases or microbial cells containing relevant reductases (Li et al. 2014; Zheng et al. 2017). In recent years, the efficient asymmetric reduction of ketones using biocatalysts as its high yield and excellent enantiomeric excess has been extensively used for the production of chiral alcohols in the chemical and pharmaceutical industries (Birolli et al. 2015; Gutierrez et al. 2010).
Carbonyl reductase (CR) or alcohol dehydrogenase (ADH) is NAD(P)H-dependent oxidoreductase which can catalyze a variety of carbonyl compounds to the corresponding alcohols (Kaluzna et al. 2005). Enantioselective CRs or ADHs have been reported from animals (Nakajin et al. 1998; Montfort et al. 2002), plants (Sengupta et al. 2015), and microorganisms (Grosch et al. 2015; Singh et al. 2009). A number of CRs or ADHs have been utilized for asymmetrically reducing carbonyl functionalities, and many excellent biocatalytic processes have been developed (Li et al. 2015). Among all the sources of CRs, microorganisms are the most important providers, and some CRs or ADHs from microorganisms have been reported with high yield and enantioselectivity (Musa and Phillips 2011). The carbonyl reductase SCRII from Candida parapsilosis could catalyze the asymmetric reduction of 2-hydroxyacetophenone to (S)-1-phenyl-1,2-ethanediol with optical purity of 100% in high yield of 98.1% (Zhang et al. 2011). Xu et al. (2015) found that an NADPH-dependent carbonyl reductase from Yarrowia lipolytica ACA-DC 50109 could efficiently convert α-chloroacetophenone to (R)-2-chloro-1-phenylethol with 99% e.e. Ni et al. (2011) heterologously overexpressed a β-ketoacyl-ACP reductase from Bacillus sp. ECU0013 in E. coli, which was used for efficient reduction of ethyl 2-oxo-4-phenylbutyrate at 620 g/L, and the e.e. of the product ethyl (S)-2-hydroxy-4-phenylbutyrate was excellent (>99%). Therefore, using carbonyl reductases to catalyze the asymmetric reduction of prochiral carbonyl compounds is an efficient and useful method for the synthesis of chiral alcohols (He et al. 2016; Cui et al. 2017; Qian et al. 2014).
The oxidoreductase from Acetobacter is mostly used to produce acetic acid, and rarely used to asymmetric reduction of carbonyl compounds to chiral alcohols. In our previous study, it was found that Acetobacter sp. CCTCC M209061 could catalyze the asymmetric reductions of several carbonyl compounds to corresponding chiral alcohols following anti-Prelog rule with excellent enantioselectivity (Cheng et al. 2014; Wei et al. 2015, 2016). In the present work, the AcCR was heterologously expressed and systematically characterized for its substrate spectrum, stereoselectivity, and the capacity of industrial application.
The Acetobacter sp. CCTCC M209061 strain was previously isolated by our group from Chinese kefir grains and stored at −80 °C. E. coli DH5α, BL21(DE3)pLysS, and plasmid pGEX-2T were purchased from Novagen. The restriction enzyme FastDigest AvaI, EcoRI, T4 DNA Ligase, DNA, and protein marker were purchased from Thermo Scientific. KOD FX polymerase for PCR was purchased from Toyobo. The kits used in the construction of the recombinant plasmids were purchased from Generay. The prochiral ketones were purchased from Sigma Aldrich or Aladdin. The primers synthesis and DNA sequencing were completed by Sangon Biotech. All other reagents and solvents were of analytical grade and used without further purification.
Genomic DNA of Acetobacter sp. CCTCC M209061 was extracted and purified using a bacterial genomic DNA Kit. Oligonucleotide primers for accr were designed according to the published gene sequence of oxidoreductase from Acetobacter pasteurianus 386B (Sequence ID: HF677570.1). The DNA fragment of accr was amplified with primer 1 (5′-TCCCCCGGGAATGGCACGTGTAGCAGGCAAGGTT-3′) and primer 2 (5′-CCGGAATTCCTTATTGCGCGGTGTACCCACCATCAAT-3′) and double-digested with AvaI and EcoRI, then the accr fragment was inserted into vector pGEX-2T, the resulting plasmid (pGEX-accr) was transformed into E. coli DH5α for amplification, and then the correct pGEX-accr was transformed into E. coli BL21(DE3)pLysS to express the recombinant AcCR. The recombinant E. coli BL21(DE3)pLysS(AcCR) were cultivated at 37 °C, 180 rpm in 50 mL LB medium (pH 6.5) containing 100 μg/mL of ampicillin. When the optical density at 600 nm (OD600) of the culture reached 1.2, the temperature was changed to 20 °C, and then IPTG was added to a final concentration of 0.4 mM. The cultivation continued at 20 °C for additional 15 h. Then the recombinant cells were harvested by centrifugation (8000 rpm, 5 min) at 4 °C and washed three times with physiological saline (0.85%), and stored at 4 °C for later use.
The oxidordeuctase activity of the recombinant AcCR was determined by monitoring the change of the absorbance at 340 nm for 3 min on the spectrophotometer (Shinmadzu UV-3010, Japan). The AcCR-catalyzed reduction of 4′-chloroacetophenone was conducted with NADH or NADPH at 35 °C in 2 mL phosphate buffer (pH 6.5 or 5.5). One unit (U) of recombinant AcCR was defined as the amount of enzyme that catalyzed the 1 μmol NADH or NADPH per minute. The NAD+- and NADP+-linked oxidations catalyzed by AcCR were conducted with isopropanol as substrate and NAD+ or NADP+ at 45 °C in 2 mL 50 mM citrate–phosphate buffer (pH 8.0). One unit (U) of AcCR was defined as the amount of enzyme that catalyzed the production of 1 μmol NADH or NADPH per minute. The reaction solution was incubated at 35 or 45 °C for 3 min, and then 20 μL recombinant AcCR (about 0.008 mg of the purified enzyme) was added to proceed the reaction.
The harvested recombinant E. coli BL21(DE3)pLysS(AcCR) cells were suspended in 50 mM citrate–phosphate buffer (pH 6.5) at the concentration of 50 mg/mL, ultrasonicated for 3 × 18 min, and then the cells debris was removed by centrifugation at 12,000 rpm at 4 °C for 20 min. The resulting supernatant was filtered with 0.22-μm filter membrane before the purification process. The general step of the purification using NGC Quest™ 10 system was as follows. First, the GST-based affinity chromatography column (5 mL) was pre-equilibrated using buffer A (4.3 mM Na2HPO4, 1.47 mM KH2PO4, 137 mM NaCl, 2.7 mM KCl, pH7.3), and then the crude enzyme was loaded on the column. After that, the loaded column was first subjected to washing with 10 column volume buffer A to remove the unbound protein fractions, then washed sequentially with buffer B (added 0.5 M NaCl in buffer A) to remove some stubborn protein, and then buffer A was used once again to lower the salinity. At last, the target protein was eluted with buffer C (50 mM Tris–HCl, 2.5 g/L glutathione, pH 8.0). The protein content was measured by the method of Bradford (1976).
The effects of temperature on the activity of recombinant AcCR were determined at various temperatures from 20 to 55 °C. For thermal stability determination, the enzyme was pre-incubated at various temperatures ranging from 20 to 55 °C, equivalent was taken at a certain time, and the residual activity was determined as described in "Measurement of enzymatic activity." The coenzyme NADH and NAD+ was used for the reduction of 4′-chloroacetophenone (5 mM) and the oxidation of the isopropanol (150 mM), respectively. The enzyme activity of the first measurement was defined as 100%.
The optimum pH of recombinant AcCR was investigated within a pH range of 4.5–9.5 using various buffer systems at 50 mM. The buffers were as follows: citrate–phosphate (pH 4.5–8.0), Tris–HCl (pH 8.0–8.5) and glycine–NaOH (pH 8.6-9.5). The pH stability was evaluated by pre-incubating the recombinant AcCR in different pH buffers (4.5–9.5) at 4 °C for 4 days. Samples were taken at a certain time, and the residual activity was determined as described above with the non-incubated recombinant AcCR as the control.
The influence of metal ions and chemical agents on the catalytic activity of recombinant AcCR was investigated by pre-incubating the enzyme with various additives in citrate–phosphate buffer (50 mM, pH 6.5) at 35 °C for 30 min. The enzyme activity was determined under the condition described above using NADH as coenzyme. The enzyme activity of the recombinant AcCR in the absence of additives was recorded as 100%.
The activity of recombinant AcCR for each specific substrate was measured by the method described above using NADH as the coenzyme. The relative activity of recombinant AcCR to 4′-chloroacetophenone was defined as 100%.
The reduction of various carbonyl compounds were performed in a 10-mL conical flask with 4 mL citrate–phosphate buffer (50 mM, pH 6.5) containing 50 mM each substrate, 0.1 mM NADH, and 150 mM isopropanol. The reactant was pre-incubated in a shaker at 35 °C for 10 min, and then 8 U recombinant AcCR was added to initiate the reaction. Samples (25 μL) were withdrawn after reaction for a certain time, and then the product and residual substrate were extracted with ethyl acetate (2 × 25 μL, 5 mM n-dodecane as internal standard) before GC analysis.
For the kinetic analysis, the initial reaction rates of the recombinant AcCR were determined under the optimum conditions. For the reduction of 4′-chloroacetophenone, the coenzyme NADH or NADPH varied from 0.05 to 0.6 mM or from 0.05 to 1.0 mM, and the concentrations of 4′-chloroacetophenone were from 0.5 to 10 mM. For the oxidation of isopropanol, the concentrations of NAD+ or NADP+ varied from 0.05 to 2 mM or from 0.5 to 20 mM. The isopropanol concentrations were from 30 to 275 mM. All the measurements were carried out in duplicate. Michaelis–Menten was used to fit the data, and the kinetic parameters of recombinant AcCR-catalyzed reduction and oxidation reactions, K m and V max values, were obtained in the fit before computing the K cat.
The organic extracts of reaction mixtures were analyzed by a GC analysis (Shimadzu Corp GC 2010). GC was performed using chiral columns HP Chiral 10B and CP-Chiralsil-Dex-CB. The initial reaction rate, the yield, and the product e.e. of those reactions were calculated as described in our previous report (Wang et al. 2013).
A 762-bp polynucleotide sequence was amplified by PCR from the genome DNA of Acetobacter sp. CCTCC M209061. The sequence was an intact open reading frame and encoded a predicted protein of 253 amino acid residues with a molecular weight of about 26.4 kDa. The GenBank accession number of the nucleotide sequence of AcCR gene is MF419650. The result of multiple sequence alignment with NCBI protein blast is shown in Fig. 1. The AcCR amino acid sequence displayed a high level of similarity to the identical proteins from other Acetobacter and related bacteria. The identities of AcCR with other proteins were as follows: 84% with 3-beta hydroxysteroid dehydrogenase from Acetobacter ghanensis (WP_059024845.1), 56% with Cyclopentanol dehydrogenase from Mesorhizobium plurifarium (CDX57396.1), 51% with R-specific alcohol dehydrogenase from Lactobacillus brevis (CAD66648.1), and 37% with short-chain type dehydrogenase/reductase from Mycobacterium tuberculosis H37Rv (NP_217373.1), respectively. The amino acid sequence alignments of the deduced polypeptides of AcCR with the short-chain dehydrogenase/reductase (SDR) proteins from GenBank database were performed. The conserved sequence (GXXXGXG) (Rossman fold motif) (Masud et al. 2011; Jörnvall et al. 2010) reported importantly of the coenzyme binding site of SDR family enzyme was found at the N-terminal 13–19 position of the AcCR. And the common active site conserved amino acid pattern SXnYXXXK was emerged at the mid-chain pattern 142–159, as well as the NNAG (89–92) sequence which has a function in stabilizing β-strands of classical SDR (Persson et al. 2003; Filling et al. 2002). Therefore, the AcCR could be classified as the SDR which belongs to a bulky dehydrogenase/reductase family.
The accr was constructed into plasmid pGEX-2T and transformed into E. coli. BL21(DE3)pLysS. SDS-PAGE analysis of the crude extracts indicated that the recombinant AcCR was successfully expressed in the soluble form with the GST tag (shown in Fig. 2). As a GST-tagged fusion protein, the GAcCR (recombinant AcCR) was purified by a single-step affinity chromatography using a GST-tagged column. As expected, the purified GAcCR was observed at the position of approximately 53 kDa (the GST tag is about 26 kDa) which was consistent with the estimated value (Fig. 2). The purified recombinant AcCR activity was 5.17 U/mg and stored at 4 °C for later use.
SDRs act on the substrate by transferring electrons from or to coenzyme. The recombinant AcCR was found to be a kind of carbonyl reductase which could use both NAD(H) and NADP(H) as coenzyme to execute the redox reactions. The coenzyme preference of the recombinant AcCR was examined by measuring enzyme activity using 4′-chloroacetophenone or isopropanol as substrate. As shown in Fig. 3, the recombinant AcCR showed redox activity with both NAD(H) and NADP(H) as coenzymes. Meanwhile, the relative activity of the recombinant AcCR with NADPH as coenzyme was only about 40% of that with NADH, which confirmed that AcCR exhibited coenzyme specificity for NADH over NADPH. In the recent reports, many biocatalysts catalyzing asymmetric reductions utilize NADPH as hydrogen donor (Leuchs and Greiner 2011; Richter and Hummel 2011; Ma et al. 2013; Zhang et al. 2015). ADHs capable of utilizing NADH as the coenzyme outperformed NADPH-dependent ones, since NADH was more stable than NADPH under operational conditions (Li et al. 2015) and economic than NADPH. The NAD(H)-preferred CRs could be advantageous for their application in industrial bioreduction system. Therefore, NAD(H) was used as the coenzyme of the enzymatic characteristic of the recombinant AcCR in the subsequent study.
Initially, the effect of varying temperatures on the activity of recombinant AcCR was explored on both reduction and oxidation reactions. The optimal temperature was found to be 35 °C for the reduction of 4′-chloroacetophenone when NADH was used as the coenzyme (Fig. 4a). For the case of the oxidation of isopropanol, the maximum activity was observed at 45 °C with NAD+ as coenzyme (Fig. 4a). The thermal stability of the recombinant AcCR was investigated at various temperatures ranging from 20 to 55 °C. As illustrated in Fig. 4b, the enzyme activity decreased slowly when the incubation temperature was less than 35 °C. The relative activity was over 50% after 36 h incubation at 30 °C, and even more than 62% after incubating for 24 h at 35 °C and over 5 h at 50 °C. Thus, the thermal stability of recombinant AcCR was excellent and much better than many reported carbonyl reductases from other sources (Xu et al. 2015; Singh et al. 2009; Luo et al. 2015; Wang et al. 2015). The enzyme exhibited half-lives of 25.75 h at 35 °C and 13.93 h at 45 °C, respectively, for the reduction and oxidation reactions.
The effect of pH on recombinant AcCR was detected in different buffer systems (50 mM) at the range from 4.5 to 9.5 for both reduction and oxidation. The maximum activity of the recombinant AcCR for reduction of 4′-chloroacetophenone was obtained at pH 6.5. As shown in Fig. 5a, the relative activity was over 90% in the reduction reaction at the pHs between 6.0 and 8.0. The optimal pH for oxidation of isopropanol was found to be pH 8.5 when NAD+ was acted as coenzyme (Fig. 5b). The optimal pH of the recombinant AcCR for catalyzing reduction and oxidation is close to that of the oxidoreductase from Gluconobacter oxydans (Liu et al. 2014).
To investigate the pH stability, the recombinant AcCR was incubated at 4 °C in varying pHs from 4.5 to 9.5 and tested the reduction activity using NADH as coenzyme. As shown in Fig. 5c, the optimal storage pH was 6.5, at which pH almost 80% relative activity remained after 96 h. Almost 70% relative activities remained at both pH 7.0 and 7.5 after 96 h, as well as at pH 6.0 after 75 h. The results showed that the recombinant AcCR had broad pH tolerant scope and excellent pH stability.
It is well known that metal ions have remarkable effects on the activity of carbonyl reductase. For example, Zn2+ is necessary for many medium-chain dehydrogenases/reductases (Zn-MDR). The Zn2+ appears in those enzymes to give the protein an additional strength, as if compensating for domain variability and evolutionary changes in the protein scaffold (Jörnvall et al. 2010). Besides, some metal ions also have negative effect on the activity of a number of enzymes. For example, Co2+, Cu2+, and Zn2+ can severely inhibit the enzyme activity of carbonyl reductase SCRII from Candida parapsilosis (Zhang et al. 2011). So it is of significantly important to explore the effects of various additives on the recombinant AcCR. As shown in Table 1, among the tested metal ions, Mn2+, K+, Fe2+, Mg2+, Ca2+, and Co2+ had slight stimulation on the recombinant AcCR, while the other metal ions had inhibitory effects to some extent. Enzyme activity was inhibited about 5 and 15% when 2 and 5 mM Zn2+ was added, and the residue enzyme activity was only 43.0 and 24.9% when 2 and 5 mM Cu2+ was added, respectively. Ag+ and Hg+ inhibited the enzyme activity completely.
The effect of chemical agents including metal-chelator, surfactants, and sulfhydryl reagents on the recombinant AcCR was presented in Table 2. EDTA had almost no effect on the enzyme activity of the recombinant AcCR, revealing that the recombinant AcCR was a metal-ion-independent enzyme. The metal-ion-independent enzyme was conformed to the characteristic of the vast majority of SDRs (Kavanagh et al. 2008; Persson and Kallberg 2013). The sulfhydryl reagents, β-mercaptoethanol and iodoacetamide, had no significant effect on the recombinant AcCR activity, suggesting that there were no essential disulfide linkages and -SH groups at the catalytic sites of the recombinant AcCR (Dako et al. 2008). Surfactants were reported to play a key role in the catalytic activity of several enzymes (Ye et al. 2009). The surfactants such as SDS, Tween-80, and Triton X-100 had obviously inhibitory effect on the activity of the recombinant AcCR, which could be due to fact that the surfactants partially disrupted the hydrophobic interactions and increased the internal repulsive forces (Adler et al. 2000).
Most carbonyl reductases can catalyze reversible reaction mediated by different coenzymes, and the substrate specificity and stereoselectivity are different towards certain carbonyl reductase (He et al. 2014; Zhang et al. 2015; Takeuchi et al. 2015; Tang et al. 2014). Therefore, it was of great interest to investigate the capability of the recombinant AcCR for catalyzing redox reaction, substrate specificity, and stereoselectivity. Seventeen of prochiral carbonyl compounds and their corresponding alcohols were explored using NADH or NAD+ as coenzyme (Scheme 1), and the results are shown in Table 3. Glancing at the results, we found that the AcCR could not only catalyze the reduction of carbonyl groups but also could complete the dehydrogenization of the partial tested alcohols. The activities of ketones reduction were higher than those of oxidation of alcohols obviously, which was beneficial to proceed ketones reduction. The oxidation of ethyl (R)-(+)-4-chloro-3-hydroxybutyrate or ethyl (S)-(-)-4-chloro-3-hydroxybutyrate was investigated, respectively, and no oxidative activity was tested. But when ethyl-4-chloroacetoacetate was used as substrate, the relative activity was tested to be 120.6%, indicating that the reaction was not reversible towards ethyl-4-chloroacetoacetate and ethyl (4-chloro-3-hydroxybutyrate. In addition, using 2-(R)-octanol or 2-(S)-octanol as substrate, the relative activity of AcCR was far from each other (107.7% vs 2.9%), which attested that the AcCR also had stereoselectivity on the oxidation of enantiomer alcohols substrates.
Chiral alcohols are one of the most useful building blocks for the preparation of pharmaceuticals. As the excellent relative activity of the recombinant AcCR towards those prochiral carbonyl-group substrates, it is of great interest to investigate the product yields and enantiomeric excess (e.e.) of the asymmetric reduction of the carbonyl compounds catalyzed by the recombinant AcCR. As shown in Table 4, AcCR exhibits a broad substrate spectrum and the enantioselectivity of this enzyme follows anti-Prelog rule which are relatively rare in nature (Tang et al. 2014; Li et al. 2015). The results shown in Tables 3 and 4 indicated that the relative activity was affected by the position of the substituents, such as the 2′-, 3′- and 4′-positions substituted acetophenone, 2′-methoxyacetophenone (16.5%), 3′-methoxyacetophenone (3.1%), and 4′-methoxyacetophenone (10.7%). High product yield and excellent enantioselectivity (>99%) were achieved when the 4′-position of acetophenone was substituted by electron-withdrawing groups such as –F, –Br, and –NO3, whereas the electron-releasing groups like -methyl had a negative effect on the activity of the recombinant AcCR. Moreover, it is noteworthy that the AcCR was more active to ethyl 4′-chloroacetoacetate among the investigated α-ketoesters, and the relative activity and yield were over 120.6 and 92.1%, respectively. The recombinant AcCR exhibited 114.4% relative activity to the 4-(trimethylsilyl)-3-butyn-2-one, and the product e.e. was over 99%, but the yield was only 62.5%, possibly because the substrate could be easily decomposed into a carbonyl alkyne and trimethyl hydroxysilane in buffer system with pH over 6.0 (Zhang et al. 2008). The product yield of reducing 3,3-dimethyl-2-butanone to 3,3-dimethyl-2-butanol was 61.7% and the product e.e was 80.7%, and if further reaction continued, the product e.e decreased markedly. What is more, unlike the acetophenone reduced to corresponding (R)-products, ethyl 4′-chloroacetoacetate and ethyl 2-oxo-4-phenylbutyrate were catalyzed to ethyl (S)-4-chloro-3-hydroxybutyrate and (S)-2-hydroxy-4-phenylbutyrate, respectively, which accorded with the anti-Prelog rule that the Cahn-Ingold-Prelog priority system was conversely exhibited when the substituted group contained higher priority atoms at its chiral center (Itoh 2014).
The kinetic constants of the purified AcCR were calculated by fitting data by linear regression to a Lineweaver–Burk double reciprocal plot. All the kinetic constants for 4′-chloroacetophenone, isopropanol, NAD(P)H, and NAD(P)+ are presented in Table 5. The results for reduction of 4′-chloroacetophenone showed that the enzyme had a much higher affinity for NADH than NADPH. Many carbonyl reductases from microorganism preferred NADPH much more, such as ADHs from Lactobacillus (Weckbecker and Hummel 2009; Leuchs and Greiner 2011), carbonyl reductase from Gluconobacter oxydans (Chen et al. 2015), and aldo–keto reductases from Candida parapsilosis (Guo et al. 2014). The NADH as coenzyme for carbonyl reductase was heavily favored for its economy and stability (Leuchs and Greiner 2011). The results for the oxidation of isopropanol (Km value 0.38 and 23.8 Vs 1.39 and 54.1) gave a proof that the AcCR was more affiliative to NAD+ than NADP+ when conducting the oxidation. Above all, the AcCR was an NAD(H) and NADP(H) dependent oxidordeuctase and more preferred NAD(H) as coenzyme in the redox reaction.
The anti-Prelog carbonyl reductase AcCR from Acetobacter sp. CCTCC M209061 was cloned and its polypeptide sequence had been predicted and analyzed to confirm that the AcCR belongs to SDRs superfamily. Then AcCR was expressed heterologously, purified, and characterized. The purified enzyme preferred the inexpensive coenzyme NADH as specific electron donor. The recombinant AcCR exhibited excellent enantioselectivity, broad substrate spectrum, and highly stereoselective anti-Prelog reduction of prochiral ketones. These results suggest that AcCR is a powerful chiral tool for the production of anti-Prelog alcohols.
Conceived and designed the experiments: WYL MHZ. Performed the experiments: PW YHC PX. Analyzed the data: WYL PW. Contributed reagents/materials/analysis tools: MHZ WYL JZ. Wrote the paper: PW WYL. All authors read and approved the final manuscript.
All authors approved the consent for publishing the manuscript to bioresources and bioprocessing. There is no conflict of interest for any of the author regarding the submission of this manuscript.
The National Natural Science Foundation of China (21336002; 21676104; 21376096), the Fundamental Research Funds for the Chinese Universities (2015PT002; 2015ZP009), the Program of State Key Laboratory of Pulp and Paper Engineering (2017ZD05), and the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.


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