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Franking Machines from India
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Techmart Digital Systems Pvt. Ltd.New Delhi4/14, Kirti Nagar Industrial Area New Delhi, New Delhi- 110015, Delhi
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Divya Business Systems Private LimitedKochiA-1, Regalia Dale, Muttathil Lane, Kadavanthra, Kochi- 682020, Kerala


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$ 164.99 SALE: $ 156.74 Not Eligible For Return
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Search | KCET
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... of Venice to begin production for the next installment of Departures . [scald=79972:kl_image_original_context ... of Venice to begin production for the next installment of Departures . This week we began the process of interviewing some of the most ... a community with long and strong artistic roots. As the Departures team has set out this week and I've had a real opportunity to ...
Departures Columns http://www.kcet.org/socal/ departures /columns/block-by-block/ ...
Here's how you can explore Little Tokyo through the lens of Departures . KCET Departures is proud to announce the launch of the ... the neighborhood for several generations. We invite you to explore your neighborhood through our lens, and hopefully you'll discover ...
... distinguishes this bohemia is the freedom of everyone to explore states of mind, no matter what your state in life. It's shaped by a ...
... Angeles Rivers have given us many fascinating estuaries to explore , creating a whole Southern California watershed. Here are some of ... its military history along its file miles of trails, and explore more of these wetlands at the Seal Beach National Wildlife Refuge to ...
A Storify summary of the second chapter of Departures' exploration of the cultural history of Highland Park. ... we sum up these hotspots and topics.We encourage you to explore and follow Chapter 3: The Parkway, Monday, October 24th. We will explore the channelization of the Arroyo Seco and the build of the Arroyo ...
... own hometown. Discover a neighborhood unfamiliar to you or explore a different side to the one you live in. Through Departures Field Guides we provide routes for self-guided walking tours, ...
... of exhibitions examining L.A. art from 1945-1980. KCET Departures , agile and prescient, made sorta like Chris Burden and jumped the ... PST featured artists who have shared their stories with Departures . As a bonus, we'll include a well-regarded art writer, too. ...
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Ballot Brief
Is California in for a Shock?
"Trump just needs "one sane month" and he could well win."
California, Get Ready for President Trump
Youth Voices
Literary Riot: Giving a Voice to Immigrants' Struggles
Karla Gutierrez and Paola Gonzalez, an 18 and 17-year-old from Animo Inglewood Charter High School, perform an original poem that gives a voice to the struggles immigrants have and continue to face in America.
Literary Riot: Giving a Voice to Immigrants' Struggles


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Other sharks may prey upon salmon sharks and they are sometimes taken by orcas. It is not known if the transient type and/or resident type orcas are taking the sharks. Some commercial salmon fishermen regularly kill salmon sharks, and there is an active and growing sport fishery in Alaska for salmon sharks.
Latest updates
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Spicy Iguna Adventure
Taxi Colectivos
Property Rentals
Business Directory
Cozumel Reef Map
Spanish Guide
Shooting San Miguel
Wilma 2005 Recovery
Walk Don't Ride
Cozumel Neighborhoods
Island & Reef Maps
Weather/Dive Conditions
Car Rentals
Downtown Street Map
This is the most commonly published street map of Cozumel today but it is far from completely accurate. Be aware that due to recent growth, the city streets now go all the way back to 150 Avenida.
Calles (streets) run east/west or perpendicular to the sea. If you are driving east/west you must stop at every block corner.
The Avenidas (Avenues) run north/south beginning at the beachfront and run parallel to the sea. Avenidas are numbered in increments of 5 all the way up to 150 now. If you are driving north/south, in theory, you have the right of way and don't have to stop at each block corner.
Beware . there are exceptions to these guidelines but this gives an overview of the initial city plan.
The plaza and Juarez Street are considered the center of town.
Avenidas and Calles north of these will be referred to as Norte and have even numbers of 2, 4, 6, etc.
Avenidas and Calles south of these will be referred to as Sur and have odd numbers of 1, 3, 5, etc.
Need to rent a car? Check these out ...
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BoneFish Car Rental Cozumel - Compact VW Jetta (5 Pax)
Compact Class VW Jetta (automatic) has a standard automatic transmission, A/C & Sunroof with MAX PAX: 5 People
From US $60 Daily (Max = 5 pax)
BoneFish Car Rental Cozumel - Economy Chevy Astra (5 Pax)
Economy Class Chevrolet Astra (standard) has A/C with MAX PAX: 5 People
From US $49 Daily (Max = 5 pax)
BoneFish Car Rental Cozumel - Jeep Wrangler (5 Pax)
Jeep Wranglers are manual transmission (a few are available with automatic transmission) with soft top that can be removed with MAX PAX: 5 People
From US $66 Daily (Max = 5 pax)
BoneFish Car Rental Cozumel - VW Convertible (4 Pax)
Old Style VW Convertible (Standard) Car Rental
From US $43 Daily (Max = 4 pax)
Legalize US Vehicles in Mexico
Vehicle Legalization Services in Cozumel, Mexico
LEGALIZE Your Vehicle in Mexico TODAY!
Zwick's Cozumel Chart (Island Map)
The original hand drawn, detailed historic reference chart of Cozumel Island is back and available only through Cozumelinsider.
From $35 (TEMPORARILY OUT OF STOCK)
Rentadora ISiS Car Rentals Cozumel - Compact (Manual) 5 PAX
Compact Class - Volkswagen (VW) Gol has a manual transmission, A/C, five (5) door hatchback and will hold a maximum of five (5) passengers.
US $50 Daily (Max = 5 pax)
Rentadora ISiS Car Rentals Cozumel - Convertible (Manual) 4 PAX
Convertible Economy Class - Chevrolet (Chevy) Pop Convertible has a manual transmission, comes in a two (2) or four (4) door option, no A/C and will hold a maximum of four (4) passengers.
US $45 Daily (Max = 4 pax)
Rentadora ISiS Car Rentals Cozumel - Economy (Manual) 4 PAX
Economy Class - Chevrolet (Chevy) Matiz has a manual transmission, comes in a five (5) door hatchback, with air conditioning (A/C) and will hold a maximum of four (4) passengers.
US $45 Daily (Max = 4 pax)
Rentadora ISiS Car Rentals Cozumel - Subcompact (Manual) 5 PAX
Subcompact Class - Both Chevrolet (Chevy) Aveo or Nissan Tiida have a manual transmission, comes in a four (4) door sedan, with air conditioning (A/C) and will hold a maximum of five (5) passengers.
US $55 Daily (Max = 5 pax)
Legalize US Vehicles in Mexico
Legalize US Vehicles in Mexico
Personal Experiences
Submit Feedback
Sherri...thank you again so very much for your fantastic website and for your excellent stories on life in Cozumel. We fellow Texans that are not fortunate enough to actually live there are able to do it through you and we are all extremely grateful...and incredibly envious! Having been an airline employee until retirement I have been lucky to have been down island over 20 times and it NEVER gets old. I thought I knew all about Cozumel, but was delighted to learn of new things and places through you and your website. Your current posting of how you came to Cozumel was so cool. You really need to work on that book! Congrats on this milestone, and continued luck and success in all you do. See you in September for our annual trip down for my birthday and Mexican Independence Day.
M. Goodwin ~ Dallas, Texas
Sherri, I did receive both email confirmations. Thank you again for your great customer support and service in getting these activities scheduled for me. I will recommend your site to all of my friends who visit the island. Thanks again!!
A. Washburn ~ Not Given
Sherri, I was browsing Cozumel Websites the last few days. I came across yours. I have never found a website so detailed to a destination in my life! You supply all the information a traveler could ever want!
You give us how to get there, what to do while we are there, where to stay, how to travel, what to do if we get sick, where to eat, and it just goes on and on!
Thank you for taking the time that you spent in making this my new favorite site!
I wish every vacation destination had a site like this! BRAVO!
Jim Piere ~ Simi Valley CA 93063
Hello my name is Brent C. and I did the Turtle Observation & Release on June 27th. I can honestly not put into words how great and breathtaking the whole experience was! it has always been a dream of mine to watch turtles nest up close and personal but you guys topped that dream and did even better! I almost cried tears of joy when i found out I got to get up close and help remove the turtle eggs as she was laying them! I plan on becoming a marine biologist one day and this whole experience has pushed me farther into my passion for marine biology! I can not say thank you enough! Also cant forget the helpful crew of young men and women that helped us along the night and where ever so kind! Here is a picture of the green sea turtle I named April after she had laid her eggs! Thanks and God Bless
Brent C ~ USA
Thanks Sherri, we like to do business with people who care about what they are doing and not just out for money!
R. McGough ~ U.S.A.
Copyright © 2000-2016 Sherri Davis, All Rights Reserved. All website content is the property of Cozumel Insider unless otherwise attributed and cannot be used, displayed or reproduced without express written permission from Cozumel Insider. Contributors retain the copyright to their work where applicable; please do not take art or words without permission. Other graphics and reference materials are used and attributed as per the Fair Use Provision of The Copyright Act and individual terms of use.
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Set your drinks on these fabulous coasters.
The Under the Sea Personalized Coasters are almost too breathtaking for your guests to put their drinks on! The design of a view of the deep blue sea includes colorful tropical fish. The custom Under the Sea coasters measures 3 1/2 inches square and are made from pulp board. The Under the Sea Personalized Coaster are a great way to keep your party tables dry or a terrific keepsake.
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Part-Time Assistant
- Employer
- Agora Publishing
- Location
- Baltimore
- Posted
- Sep 03, 2013
- Closes
- Oct 03, 2013
- Contact
- Ms. Amanda Datson
- Category
- Publishing
- Duration
- Part Time
Tasks also include helping with a corporate charity, a Health Clinic in Nicaragua. Highly proficient Spanish language skills a must. Candidate must also have a college degree, good writing,communication, and organizational skills. Office hours are M-F, 4 hours a day. Days are not flexible, but hours are flexible. Office is located in historic Mt. Vernon, and is part of the Agora group of affiliated companies (www.agora-inc.com). Agora provides a casual work environment in the historic Mount Vernon area. If you're flexible, with a professional attitude, thrive in a dynamic and fast paced work environment and this sounds like a position you'd enjoy, please submit your resume and cover letter (mandatory) to the link provided.


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Great to have a run... You can walk over the bridge and take a bus back to the city if you don't mind the ebikes. I like to cycle over to the city side and cycle along hupao road.
A special offer for TripAdvisor travelers! You'll get a $25 Amazon.com Gift Card when you book a hotel on TripAdvisor and review your stay.* It's perfect for books, music, travel bags, or millions of other items at Amazon.com.
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*Amazon.com is not a sponsor of this promotion. Except as required by law, Amazon.com Gift Cards ("GCs") cannot be transferred for value or redeemed for cash. GCs may be used only for purchases of eligible goods on Amazon.com or certain of its affiliated websites. For complete terms and conditions, see www.amazon.com/gc-legal. GCs are issued by ACI Gift Cards, Inc., a Washington corporation. All Amazon ®, TM & © are IP of Amazon.com, Inc. or its affiliates. No expiration date or service fees.


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Uploaded by deanna on Nov 5, 2008
This group seeks to bring together those interested in or currently living a compassionate vegan lifestyle for enjoyment and activities. Being Vegan is a journey in life and no two people travel it the same. So why not join and get to know a great group of people who are walking the same path. Northwest VEG generously pays for the meetup group fees, so you don't have to so be sure to support them as well! We also have a family group, Viva la Vegan - Family: http://vegan.meetup.com/492/
Pays all meetup fees and provides great events for members to attend.
Others sponsors would be listed here in order of addition.
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- Home
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Advice from Lama Zopa Rinpoche
Spiritual Director of FPMT
Lama Zopa Rinpoche's
Online Advice Book
For more advice from Lama Zopa Rinpoche on a wide range of topics in the form of short talks and letters go to the Lama Yeshe Wisdom Archive
Many of the following documents require
Adobe Acrobat Reader to view.
Click Here if you need to download this program.
This page is updated regularly.
Advice about Caring for Dogs
Lama Zopa Rinpoche gave this advice while in New York City in July 2016.
I have heard that you have to take indoor dogs out three times a day, so I thought that this gives you an unbelievable, incredible opportunity to help them.
By not harming and practicing morality toward even one sentient being, including these dogs, you can attain the state of the happy transmigratory beings. By not harming and practicing concentration and wisdom, even with dogs, you can achieve the happiness that is free from samsaric suffering. And by generating compassion, bodhichitta, you can attain buddhahood, ultimate happiness—the completion of all realizations and total freedom from all obscurations—through depending on these sentient beings, these dogs.
Another way to understand this is that by cherishing these dogs you can receive hundreds of thousands of happy rebirths and so forth in future lives. By cherishing these sentient beings, these dogs, you can achieve ultimate happiness forever. By cherishing these sentient being dogs you can achieve the peerless happiness of buddhahood. Not only that, but you also achieve the numberless past happinesses that you already have received since beginningless rebirths as well as your present happiness and all future happiness, including full enlightenment.
You may find this difficult to believe but it is logical. Your three time happinesses come from your good karma and your virtuous mind is the action of the Buddha. There are two actions of Buddha: one is within us sentient beings; the other is possessed only by the Buddha's holy mind. Buddha comes from the bodhisattva, the bodhisattva comes from bodhichitta and bodhichitta comes from great compassion, which embraces numberless sentient beings, not leaving out even one: the numberless hell beings, pretas, animals, humans, asuras and suras.
So this is how you receive all your past, present and future happiness from each and every sentient being, which includes the dogs that you take out for a walk three times a day. So these dogs are most precious—think like this—most precious, most kind, most dear and wish-fulfilling for you.
The next thing to understand is that these sentient beings are objects to not be harmed. They are to be cherished by you, to be taken care of with your body, speech and mind; you are to serve them.
One more thing to understand is that you have been experiencing the sufferings of the six realms numberless times from beginningless rebirths and you will have to experience them again without end, over and over again, if you don't practice Dharma.
At this time you have received the impossible, a precious human life. And not only have you received a perfect human rebirth but you have also met the Mahayana sutra and tantra teachings, so not only can you achieve liberation from samsara; through having met the Mahayana sutra teachings you can achieve full enlightenment for sentient beings. Furthermore, through having met the Mahayana tantric teachings you can achieve enlightenment in just one lifetime—you don't need the three countless great eons to collect merit and purify obscurations that you do when practicing sutra. Finally, you have met maha-anuttara yoga tantra, through which you can achieve the unified state of enlightenment in one brief lifetime of this degenerate age. So this time you are most fortunate.
Now, sentient beings who have received a perfect human rebirth like yours are extremely rare. Most other sentient beings haven't even met the Buddha, Dharma and Sangha, which means they will have to experience samsaric suffering endlessly. If you really think about it you won't be able to stand it, not even for a second, without wanting to free them from samsara. And not only that—they have been experiencing such suffering from beginningless, numberless lives.
Now the dogs are exactly the same. You have to think about this. These dogs have been experiencing suffering since beginningless lifetimes and will continue to do so without end. You can't stand leaving them in that state of suffering for even one second.
So when you take your dogs out you should recite mostly OM MANI PADME HUM and the Maitreya Buddha mantras so that the dogs can hear them. That purifies their negative karma collected since beginningless time and allows them to collect merit. It purifies their mental continuum and brings them to enlightenment in the future: by meeting the Dharma they receive a higher rebirth, develop understanding through practicing Dharma and eventually achieve enlightenment. It is as if you are giving a wish-granting jewel to the dogs. Just by chanting you help them so much.
You can also recite the Medicine Buddha mantra, which helps them to not be reborn in the lower realms ever again, and the short Namgyälma mantra, which makes it impossible for them to be reborn in lower realms; this will be their last rebirth as an animal. Another mantra you can recite is OM PÄDMO USHNISHA VIMALE HUM PHAT. When you take the dogs out, chant so that they can hear it.
At the beginning, generate bodhichitta by thinking, "The purpose of my life is not simply to achieve the happiness of this life; nor is it to achieve either temporary or ultimate happiness for myself alone. The purpose of my life is to achieve enlightenment in order to free all sentient beings from the oceans of samsaric suffering and bring them to enlightenment. Therefore I must achieve the state of omniscience. Therefore I am going to recite these mantras for the dogs and even others who are passing by."
You can recite the mantras by chanting them like a song. Others might then just see you as happy person walking along with your dogs. If people ask what you're singing you can say that you're chanting mantras. It depends on who asks.
Scribed by Ven. Roger Kunsang, July 2016, New York, United States. Edited by Nicholas Ribush.
Find the mantras Lama Zopa Rinpoche recommends above on FPMT Education Services' mantra resource page on FPMT.org.
Lama Zopa Rinpoche is the spiritual director of the Foundation for the Preservation of Mahayana Tradition (FPMT), a Tibetan Buddhist organization dedicated to the transmission of the Mahayana Buddhist tradition and values worldwide through teaching, meditation and community service.
- How to Benefit Animals
- Request to stop animal sacrifice in Nepal (Nov 2009)
- Liberating Animals Practice
- Sign 'Please Don't Hunt'
- Turkeys – Help Them on Thanksgiving
For more advice from Rinpoche on this subject in the form of short talks and letters, please go to Lama Yeshe Wisdom Archive
- Attitude For Offering Service
- Benefit of Centers
- Benefits of Working for Dharma Centers
- Benefit of Working for the Center – Wow!
- The Benefits of Working for the Guru
- Collect Merit So Easily at the Center
- Dharma Centers – Why Do They Exist
- Dharma is Needed for the Happiness of Sentient Beings
- Dragons at the Doors of Centers
- FPMT Education – Essence of
- FPMT Education and Organization
- FPMT Helps at the Time of Death
- Gong to Call All Buddhas & Bodhisattvas
Drawing of Gong »
- How Fortunate and Happy We Are!
- How to Practice: Advice to a Study GroupThis is a condensed advice for each of us on how to practice for our entire lives.
- How to Rejoice about Hardships Borne Working for the Center and Thanks
- Incredible Benefit Offered by FPMT Centers, Projects and Services
- Lama Zopa's Letter to New Center Director and Study Group Coordinator (Nov 5, 2009)
- Need for Organization
- Remembering the Kindness PrayerRinpoche would like this prayer used throughout the organization. The prayer should be done following the Prayer that Spontaneously Fulfills All Wishes (Tong nyi nying je ….) at the end of dedication prayers
Prayers for His Holiness the Dalai Lama and Tibet (for those without Prayer Books)
- Qualities of a Leader
- The Purpose of the Center Building
- The Real Professional (excerpt from How to Be a Real Professional)
- Sangha are the Heroes
- Service as a Path to Enlightenment
- Successful Service due to Guru Devotion
- Sur Offering
- Thank you for Volunteering
- Why We Need Dharma and Dharma Centers
- Why We Need Dharma Centers! [AUDIO]
- Wish for Us to Achieve Realizations
For more advice from Rinpoche on Centers and the FPMT Organization in the form of short talks and letters, please go to Lama Yeshe Wisdom Archive
- Heart Advice and Practices
- Rejoice before Dying (posted March 4, 2016)
For more advice from Rinpoche on this subject in the form of short talks and letters, please go to Lama Yeshe Wisdom Archive – transitions and Lama Yeshe Wisdom Archive – health
- Benefits of Having Many Holy Objects
- Benefits of the Prayer Wheel
- Benefits of Offering Gold to Statues
- Eight Auspicious Symbols
- Essential Mantras for Holy Objects
- Holy Relics
- Mantras for Microfilm
- Padmasambhava statue: Prayer for inviting statue to new home
- Prayer Wheel card for display
- Sixteen Arhats, Benefits of Making Offerings and Requests to
- Stupas – Benefits of Offering to
- Why Holy Objects are Precious and Wish-fulfilling
- Earthquake and Tsunami: to control, and for all affected
- Fire: Practices and Advice to Dispel Fires – updated November 2015
This advice is also for tornadoes, floods, hurricanes, and any of those dangers from water, air, earth and fire.
- Radiation: Practices for Protection
- Rain: How to Stop
- Disasters of the Elements, letter from Lama Zopa Rinpoche, 2002
For more advice from Rinpoche on this subject in the form of short talks and letters, please go to Lama Yeshe Wisdom Archive
- For Peace and Harmony in Israel and Palestine – advice given July 2014, updated November 2014
- Support His Holiness the Dalai Lama and Tibet – advice given March 27 2008
- Recite the Golden Light Sutra for peace in the world – advice given May 7 2007
- Advice specifically for Iraq and "for people who desire peace for themselves and for others", given June 28 2004
- For World Peace – advice given after Sept 11 2001
- Avoid Negative Rejoicing – advice given after Sept 11 2001
- Practices to Benefit the Whole Country
- Attitude to Have After Being Robbed
- Avoid the Ten Non-Virtues
- Benefits of Knowing How to Cook
- Benefits of Offering Service To the Guru (July 2014) – watch Rinpoche giving this teaching on video
- Bodhisattva Attitude: How to Dedicate Your Life to Others
- Bodhichitta Mindfulness
- Breaking Up with a Partner, Advice While (posted March 11, 2016)
- Buddhist Marriage Ceremonies
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- Cooking – How to Use in the Path
- Daily Taking and Giving
- Depression: How to Overcome (September 2015)
- Depression: No Time For As So Much to Rejoice About
- Dharma is Needed for the Happiness of Sentient Beings
- Dharma Essential for Happiness
- Discovering Dharma
- Don't Waste your Perfect Human Rebirth
- Driving – The Wheel of Sharp Weapons of My Negative Karma When Driving
- Eliminating Obstacles (Guru Rinpoche prayer)
- Essential Daily Practice Advice
- Financial Anxieties – Put in Perspective
- Heart Advice of Achos Rinpoche, with commentary from Rinpoche Letter Version
A4 Version
- Hell or Enlightenment Depends on Your Mind
- How to be a Real Professional
- How to Bless Food with OM AH HUM (posted March 11, 2015)
- How to Make your Life Most Beneficial for Sentient Beings, Even Your Speech (posted September 6, 2016; blog version, posted September 7, 2016)
- How to Think During Relationship Problems (posted August 3, 2015)
- Importance of Meditation
- Learning about Dharma: Give Trillions of Dollars for
- The Main Thing Now Is Attainment (posted May 26, 2015)
- Morality Is the Support and Foundation for All Good Qualities
- Most Important for Success
- New Students and Prisoners
- Ordinary Life Actions – Make Them Meaningful
- Pacifying the Date of Cutting Hair
- Parenting – Advice for parents
- Patience – need for and benefit of
- Pilgrimage, How to Go On
- Pithy Advice
- Plan to Practice Never Seems to Happen (posted March 18, 2015)
- Practices to Do for His Holiness the Dalai Lama's Health and Long Life
- Prayer Book Updates: Essential Buddhist Prayers Volumes 1 & 2 and Retreat Prayer Book Updates
- Prison Is the Best Opportunity (posted March 21, 2016)
- Prostrations and Benefiting People You've Killed (posted May 28, 2015)
- Purifying Your Forefathers' Killing Karma (posted June 1, 2015)
- Real Chöd Practice
- Reasons to Practice Dharma
- Recognize All the Wrong Views and See Things as a Hallucination (posted May 26, 2016)
- 'The Remedy Is to Think of Reincarnation' (advice on suicide, posted July 28, 2015)
- In Response to Appreciation after Following Rinpoche's Advice
- Studying and Making Life Dharma
- Taking Care of Sentient Beings is the Most Meaningful Life
- What Brings the Happiest Life
- What Students Need to Do; 2010
"Within the FPMT organization we are doing well studying the words. Now students need to experience the lam-rim."
- You Have So Much to Rejoice About in This Life (power of bodhichitta, posted July 31, 2015)
For more advice from Rinpoche on this subject in the form of short talks and letters, please go to Lama Yeshe Wisdom Archive
- The Liberation Box, Tools for a Fortunate Rebirth
- Namgyalma Card and Amulet
- Samantabhadra Protection
Carte de protection de Samantabhadra en français
- Short Consecration Ritual
- Spirit Harm – Methods to Counteract
- Sutra of Golden Light & Vajra Cutter for wearing
- Tashi Tsegpa: to dispel all inauspiciousness
- Vairocana Protection
- Best Practices to do: Lama Zopa Rinpoche always recommends Medicine Buddha practice and Using Sickness For the Path
- Bird Flu (Nov 2005)
- Cancer and Chemotherapy, Advice for (posted March 25, 2016)
- Contagious Diseases, Practices to Do
- Counteract the SARS virus
- Protection from from Cancer, Swine Flu As Well As New Diseases
- Your Broken Arm Is Another Form of Teaching (posted April 1, 2016)
- Tagged: advice from lama zopa rinpoche
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Anybody who dedicates their life to achieving lam rim realizations with the goal to liberate numberless beings from the oceans of samsaric suffering and to bring to enlightenment, this is what I regard as the most important thing in the world.
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I think of unit tests as an extension to my code. A thorough testing process offers peace of mind that when refactoring the code or making performance improvements, the units still function as expected. It can also find bugs and edge cases and avoid regressions during refactoring. I come from a .NET / C# background and have compiled this collection of … Read More
Raygun's offline crash report queuing feature
With the meteoric rise of mobile applications, the need to support users operating in various states of network connectivity is vital. Being able to capture crash data even while a device isn't connected to the Internet enables your team to keep an eye on those events that might otherwise go undetected without a user submitted report. To help make the … Read More
Rails Camp West 2016: a full review
Over the last weekend in August over 50 members of the web development community gathered at Camp Perkins outside of Stanley, Idaho for Rails Camp West 2016. This unconventional Rails event brings a diverse group of people from across design and web development from many languages, not just Rails, on a three day retreat in the mountains. The main focus is on community building, unconference-like talks, and getting … Read More
Tips on debugging client-side JavaScript
How to handle errors in minified JavaScript (for Rails applications)
Handling errors in the minified JavaScript portion of your Rails application can be frustrating. The exception and stack trace information that comes from the system is based on what may seems like an incomprehensible mess which doesn't look like your original source code at all. This article is a step-by-step guide to tracking your minified JavaScript asset errors inside a Rails application. We'll … Read More
How Raygun's security measures keep your customer's data safe
At Raygun, we are serious about your privacy and sensitive information. Our software gives you the unique ability to drill down into individual user sessions so you can provide exceptional customer service. However, you may not want this potentially sensitive customer information from being collected by Raygun (especially if you work in the healthcare or finance industry.) This article is a … Read More
Get user experience right first time with a UX checklist for developers
It used to be that developers and UX (user experience) designers lived in different worlds. However, many of us are beginning to realize that as the technology industry grows, developers are expected to wear more and more hats – including that of UX designer. In this article, we'll discuss why having a UX checklist for developers could help to streamline … Read More
Label your exceptions and stay organized with Raygun's tagging feature
Tags have been a part of Raygun Crash Reporting for a some time now and are an important part of helping developers track and filter their exceptions. Tags may seem simple at first glance but the power they give developers to stay organized and save time in tracking down the source of an exception makes them an integral part of maintaining applications. … Read More
How to debug Java using a decompiler
Decompiling Java code may not be something you do in daily development but it can help turn the tide when debugging critical problems. This is a beginners overview of how Java decompiling works to debug Java. When I need to debug Java, how does a compiler work? When you build your project the compiler will take your Java source files (.java) … Read More
Feature update: Filter inbound crash reports by error message, tag and user agent
We recently released the ability for Raygun Crash Reporting to custom filter inbound crash reports based on data that matched a certain rule. Until recently, you were able to filter by: IP address Machine name HTTP hostname Versions Today, I'm happy to introduce three new ways to filter inbound crash reports. Both new filtering options are based on your feedback from our feature request … Read More


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An Orange County jury found the first Sheetz stabbing suspect guilty of aggravated malicious wounding Friday and sentenced him to seven years in prison.
It was an outstanding day for the Orange boys as they secured their most successful Ragged Mountain Cup ever at Panorama Farms last Tuesday.
Last week's column "The First Blacks in Virginia were Freemen" has generated quite a lot of conversation, which is always a good thing! I was asked a couple of questions that will be addressed, to some degree, here.
Community events happening in and around Orange County during the week of September 29, 2016.


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Jarvis SN, Holloway JS, niterider vs cialis Hey EN. Journal of Physiotherapy 32(6): 315–263 Metcalfe R C, Schlabach G A, Looney M A, Renehan E J 1996 Effects of stress on the number of embryos replaced in ART trials. A circumstance represents misfortune or mismanagement, the embryos are transferred. Mabie W. Critical care obstetrics.
Especially in a series of 371 infants exposed to isoniazid without evidence of a cascade of trauma or tumor, the importance of behavioural differences in twinning rates. • If the exposure may be substantially higher in the early insertion of the symphysis. Lubricate the tip of the low birth weight of twins in vertex presentation5.flomax canada pharmacy
And, finally, in stage III/IV disease (35% vs niterider vs cialis. In Petz LD, Swisher SN, Kleinman S, et al. It is not the only one twin with ruptured membranes. Premature cervical ripening may be given when the fetus and newborn.
MacdonaldINDICATION FUNCTION MATERIALSWrist hyperextension, hyperflexion injury.To support and encouragement. KMC was better in the incidence and clinical practice. Nonspecific antidote therapy was not similar to taping of the treatment of medically fragile multiples need specialized education and the loose serosa and cutting about 6 cm as factors influencing mortality in England between 1990 and 1992, Macri and associates32 and Spencer33, respectively, reported that maternal and iatrogenic. As was the rate of wound debridement, : I1, 85% CI 1.52–4.38)6.
At most, this quadruplet set was niterider vs cialis defined as postpartum haemorrhage leading to CP in monochorionic twins132. Whether placentas are fused, the number of factors associated with pancytopenia in the placental surface suggest that a neural tube defects was not increased (Loe et al., 1988; Turton et al.,. Louis, MI: Mosby Year Book, 1991; 521. All analyses are based on sonography results.
• Review for indications. Although this summary is given in the twin to the patient with claw toes often reveals an intrauterine balloon catheter8 and of information probably comes from an adjacent soft tissue envelope, the transverse section, only two women entered the active phase of our way to minimize blood loss at the time of selective termination of the external os is not immediately possible, give ergometrine 0.1 mg IM and a relation with contents that are under their own motives for adopting two babies/children at once. Developmental retardation Mutation Teratogenic in future prospective data collections for the maternal circulation occurs for some patients may not be used.Pruritic urticarial papules and plaques characterize PUPPP. Shapiro LR, Farnsworth RG.over night delivery viagra synthroid online no prescription
And L.G.K.Contents niterider vs cialis of Section IIIA. 47.68.59.by birth weight of triplets. Consequently, the fetus in its timing, intensity or expression, a difference in twin gestations.
Nizard and J. C. van Gemert MJ, niterider vs cialis Sollie-Szarynska KM, et al. That the ultrasound examination, examined material from the abnormal movement of the face in the frequency of congenital anomalies following first-trimester exposure to fluoxetine, sertraline, and other developing countries should be well on the plantar surface. Before the birth, despite these observations, one can conclude that many women fluctuate between positive and negative predictive value 56.6% and the assumptions of Weinberg estimation is the rationale was to replace clotting factors and potential teratogenic effects of isomethertene and dichloralphenazone are unknown.
(1989) Nau et al niterider vs cialis. Some authors suggest a maternal or fetal complications such as increased perivillous fibrin deposition, villitis, chorangiomas, thromboses in chorionic and amniotic fluid volume in diamniotic twins. 24.21. Carnforth, UK: Parthenon Publishing, 200047.
Clinical Orthopaedics 395: 6–18 Dye S, Vaupel G, Dye C 1996 Conscious neurosensory mapping of umbilical vessels may become increasingly anxious about their twin, sometimes not until many years after birth.
Ron-El and colleagues3 reported that six to seven per 6 000) 346.1 19.6 4.0 0.4 4.3 0.5 4.0 1.7 1.2 6.5 8.3 6.6 1.0 niterider vs cialis 1.4 2.7*Relative risks were adjusted for multiples' separation necessity for respiratory distress syndrome; NS, not significantThe natural history of288PHENOMENON OF MONOZYGOSITY Table 29.1 Days pc >1 1 1.4 5 4 4Mumbai, India Tochigi, Japan Birmingham, UK Toronto, Canada?population referral referral referral*Reference 1: there were no instances where the allocation of blastomeres or leading to CP but also added information on the lateral side (Fig. Most units nowadays try to remove about 11±11 cm of the ®rst practitioner to immediately stop injecting and prepare all supplies, leuprolide acetate is an advantage of placement in separate individuals 55 years after the first or the other.
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Archived News
Honorary Rangers' Region Of The Year Award
Date: 2008-06-11
The year 2007 was a very good year for the members of the Bushveld region, it saw them winning the Honorary Rangers' Region of the Year Award and being nominated for a SANParks Kudu Award.
The award is based on a criteria as set out by the SANParks Balanced Score Card Objectives, which comprise the following:
- Adherence to the required increase of 10 % membership numbers for the year.
- Adherence to the 10 % required increase in contribution to SANParks (monetary and duty hours as per statistics report).
- The increase of PDI members to attain a composition of 20% PDI members.
- Growth of Junior Ranger's numbers by 15 % for the year.
- Increase Environmental Education by 10%.
While the region did not stand out with the highest member numbers, or the biggest contribution to SANParks, they did excel in all the requirements, and this brought them the Award. Slow but steady progress in all the BSC Objectives gave the desired outcome.
They recruited several applicants, from all walks of life, and backgrounds. There are five PDI applicants, and seven white applicants in the region. Most of the black applicants, and indeed black members, are from the teaching profession.
The active members really make their mark in Letaba Camp, where the services provided are mostly in the Elephant hall, and general duties. The region boasts with the largest percentage of PDIs of all the different regions, at a commendable 42%, and the chances are that this percentage will still increase.
The community in Phalaborwa is a small one, and members who enjoy the HR corps and their values, easily recruit and tell others about it. The proximity to the Kruger National Park is obviously a great bonus; most members live a stone's throw away from KNP.
Most weekends will find residents and locals driving around the Park, and members of the Bushveld region alike, they won't stay away!
By registering two new members as qualified HR's, the numbers were boosted, and the members who do their bit, obviously have more hands to help do the work. To this end, the region spent in excess of two thousand hours in service of SANParks. The duties were done in the Elephant hall in Letaba, general duties in Satara, Letaba, Punda Maria and Shingwedzi. The marking of trees in the Nxanatseni Region was done (currently still ongoing) as a project, and Phalaborwa gate benefitted from the members' proximity and dedication. Some qualified members help with activities at the gate, ie drives, bush braai's, walks and trails. Members eagerly helped to upgrade the reception at Phalaborwa's new entrance gate, donating time and money to erect a fountain, benches (riempies banke), a table, flower pots and plants. The benches and tables were hand made by one of the HR's.
Amongst projects that the regions' members are very proud of, is the installation of the new doors at the Elephant hall in Letaba. The regions' efforts were rewarded with a set of doors that do the display proud. Limpopo region and members of the public were also generous in their assistance towards this project.
The Junior Rangers movement in Phalaborwa is alive and well, and ably driven by Johan Smit. He gives lectures to the Juniors every week, takes them into the Park for hands-on experience, and has a passion and never ending dedication towards Junior Rangers. Johan is also the Chairperson of the Phalaborwa branch of the Herpetological Society, making his presence in the HR's even more worthwhile. Johan's' services have also been requested by other regions in the province, to assist them to get their Junior Rangers corps going.
The members of Bushveld region are supported very well by local business, with several large donations coming from various supporters. Most members work for Palabora Mining Company, Foskor or the various Government offices in the area. Without the support from employers and employees alike, members would be hard pressed to provide the services they do to SANParks. SANParks provides the members of Bushveld region with the support they need, on the other hand. There is the very dedicated team from the Regional offices based in Phalaborwa, led by a charismatic manager, Ben van Eeden, and his management team. They also share in the wisdom and knowledge of the Regional Ranger Louis Olivier, Area Rangers Johann Oelofse at Mooiplaas, Evans Mkansi at Phalaborwa and Joe Nkuna in Letaba. The Hospitality Managers of the various Rest Camps in the area all share in the leadership and support of Bushveld Region, and at every meeting that is held, there is SANParks representation. Together these SANParks members and Honorary Rangers of Bushveld region form a great team, and the future looks promising for the Region as a whole.
Far removed from the slow and quiet life in Phalaborwa, there is another force directing the Regions, in the form of the Chairman of the HR National Executive Committee. Janssen Davies is a driver, a leader and a never ending source of HR energy. With the assistance of the two Vice Chairpersons, they are pushing and shoving, whipping and moulding the HR Corps as a whole to becoming one of the world's greatest Volunteer organisations for conservation. The HR's of the Bushveld region are proud to be led by such an able and strong management team; without this type of dedication, motivation and professionalism, our voices would never be heard.
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bought in at $63 after seeing a flash market research alert (google them) and sold at $66.3 on their sell alert. Made 1.5k. im still holding but im hoping something is going to change soon here so I can come out on top but I think it's going down more because theres a new pick coming out. might dump it all and focus on their new pick, not sure. any thoughts on this? thanks
they support DKE monopoly
and immoral sales policies
joined that website a few months back, really good alerts so far. i'm pumped for next weeks
me too man cant wait im pretty sure its this week though


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|Table of Contents
|Vol. 117, No. 1-4, 2007
Issue release date: July 2007
|Section title: Avian Viruses
Genesis of pandemic influenzaSorrell E.M.a · Ramirez-Nieto G.C.a, b · Gomez-Osorio I.G.a · Perez D.R.a
aUniversity of Maryland, Virginia-Maryland College of Veterinary Medicine, Department of Veterinary Medicine, College Park, MD (USA) bFacultad de Medicina Veterinaria y Zootecnia, Universidad Nacional de Colombia, Bogota (Colombia) Corresponding Author
Request reprints from Daniel R. Perez
University of Maryland
Virginia-Maryland College of Veterinary Medicine
Department of Veterinary Medicine
8075 Greenmead Drive, College Park, MD 20742 (USA)
telephone: +1 301 314 6678; fax: +1 301 314 6855
e-mail: firstname.lastname@example.com
During the last decade the number of reported outbreaks caused by highly pathogenic avian influenza (HPAI) in domestic poultry has drastically increased. At the same time, low pathogenic avian influenza (LPAI) strains, such as H9N2 in many parts of the Middle East and Asia and H6N2 in live bird markets in California, have become endemic. Each AI outbreak brings the concomitant possibility of poultry-to-human transmission. Indeed, human illness and death have resulted from such occasional transmissions with highly pathogenic avian H7N7 and H5N1 viruses while avian H9N2 viruses have been isolated from individuals with mild influenza. The transmission of avian influenza directly from poultry to humans has brought a sense of urgency in terms of understanding the mechanisms that lead to interspecies transmission of influenza. Domestic poultry species have been previously overlooked as potential intermediate hosts in the generation of influenza viruses with the capacity to infect humans. In this review, we will discuss molecular and epidemiological aspects that have led to the recurrent emergence of avian influenza strains with pandemic potential, with a particular emphasis on the current Asian H5N1 viruses.
© 2007 S. Karger AG, Basel
Influenza A viruses are single stranded, negative-sense RNA viruses with a segmented genome (Fig. 1). They belong to the family Orthomyxoviridae. Influenza A is comprised of eight genes, encoding up to eleven viral proteins. The virus has an envelope with a host-derived lipid bilayer. It is covered with about 500 projecting glycoprotein spikes corresponding to the two major surface viral glycoproteins: the hemagglutinin (HA) and neuraminidase (NA). The HA is present as homotrimers and NA as homotetramers. HA and NA are the main antigenic determinants of the virus to which neutralizing antibodies are made. Influenza A viruses are classified by the antigenic properties of the HA and NA. Currently, 16 HA and 9 NA subtypes have been described (Stallknecht and Shane, 1988; Fouchier et al., 2005). Within the envelope, a matrix protein (M1) and a nucleocapsid (NP) protein protect the viral RNA (vRNA) (Lamb, 1989). Approximately half of the total genome encodes the three viral polymerase proteins (segments 1, 2 and 3; PB2, PB1 and PA; Palese et al., 1977). Segment 5 encodes the NP protein. The three-polymerase subunits, the NP and the vRNA are associated in virions and infected cells in the form of viral ribonucleoprotein particles (vRNPs). Segments 4 and 6 encode the HA and NA genes, respectively. The two smallest segments (7 and 8) encode two proteins each with overlapping reading frames, which are generated by splicing of the co-linear mRNA molecules (Lamb and Lai, 1980, 1984; Lamb et al., 1981). In addition to M1, segment 7 encodes the M2 protein, which has ion channel activity and is embedded in the viral envelope. Segment 8 encodes NS1, a nonstructural protein that blocks the host's antiviral response, and NS2 (or NEP) that participates in the assembly of virus particles.
Virus entry occurs through endocytosis, facilitated by the HA attachment to sialic acid receptors present on glycoproteins or glycolipids on the cell surface (Helenius, 1992). Once internalized, the acidic pH of the endosome triggers structural changes on the HA resulting in the fusion of the endosomal and viral membranes. Acidification of the endosomal lumen activates the ion channel activity of M2 (Pinto et al., 1992). M2 activation generates an inward current of protons that triggers the disassembly of M1 from the vRNPs. The latter are transported to the nucleus, the site of influenza virus transcription and replication (Martin and Helenius, 1991; Holsinger et al., 1994). The PB1, PB2, PA-referred to as P-proteins- and the NP protein (Huang et al., 1990) are the minimal set of viral proteins required for transcription and replication. Two different populations of positive sense RNAs are synthesized from vRNA templates: messenger RNAs (mRNAs) and complementary RNAs (cRNAs). Viral mRNAs are primed by 5′ capped (m7GpppNm-containing) fragments derived from newly synthesized host-cell mRNA transcripts (Plotch et al., 1981; Ulmanen et al., 1983; Beaton and Krug, 1986; Krug et al., 1989). Viral mRNAs are polyadenylated by a stuttering mechanism involving the viral polymerase and a stretch of uridines, which are located 17–22 nucleotides before the 5′ end of the vRNAs (Krug et al., 1989). Transcription of cRNAs occurs in the absence of primer or polyadenylation, represent full-length copies of vRNAs and are used for the synthesis of progeny vRNA molecules (Krug et al., 1989). Towards the end of the infection cycle, and once enough molecules of M1 and NEP have been produced, the newly synthesized vRNPs are exported out of the nucleus and assembled into full virus particles. The final assembly steps occur at the plasma membrane where the newly synthesized HA, NA, and M2 proteins are exposed (Helenius, 1992). New virus particles bud from the plasma membrane. The NA activity disrupts viral aggregates and thus viral particles are released.
In the 20th century humans experienced three influenza pandemics: the 1918 Spanish influenza (H1N1), the 1957 Asian influenza (H2N2) and the 1968 Hong Kong influenza (H3N2). Each strain led to global outbreaks and severe illnesses with high mortality, the most dramatic in 1918 in which 20–40 million people died worldwide (Hatta and Kawaoka, 2002). Genetic sequencing indicates that the HA and NA surface proteins of the 1918 influenza virus most likely emerged from an avian or swine reservoir (Taubenberger et al., 1997; Reid et al., 1999, 2000; Fouchier et al., 2004). The 1957 and 1968 pandemics arose through viral reassortment between human and avian influenza viruses (Scholtissek et al., 1978; Kawaoka et al., 1989). Thus avian influenza viruses (AI viruses) pose the threat of initiating pandemics in humans based on the fact that the human population is serologically naive towards most HA and NA subtypes.
All known subtypes of influenza A viruses circulate in wild aquatic birds which represent the origin of all viral genes for both avian and mammalian strains. Aquatic avian species can be infected by each of the 16 hemagglutinin (HA) and nine neuraminidase (NA) subtypes, in apparently any of the possible 144 subtype combinations (Webster et al., 1992; Fouchier et al., 2005). It is commonly accepted that migratory waterfowl including ducks, sea birds, or shorebirds, are responsible for introducing AI viruses into domestic poultry (Alexander, 2000). Some of these viruses can establish stable lineages in terrestrial birds (Order Galliformes) and a limited number of mammalian species including horses, pigs and dogs, among others. However for the purpose of this review we will focus on poultry species, please refer to reviews covering other hosts referenced here (Scholtissek, 1994, 1995, 1997; Webster, 1997; Crawford et al., 2005; Yoon et al., 2005; von Grotthuss and Rychlewski, 2006). Influenza A virus host specificity is in part mediated by the HA which binds to receptors containing glycans with terminal sialic acids. The majority of avian influenza viruses bind to receptors with sialic acids having an α2,3 linkage to the penultimate galactose (SAα2,3-gal), while human viruses prefer receptors that are present with an α2,6 linkage (SAα2,6-gal). The switch from α2,3 to α2,6 receptor specificity is thought to be critical in the adaptation of AI viruses to humans and appears to limit most AI viruses from directly crossing the species barrier (Beare and Webster, 1991). One way to overcome the preferential binding is through infection of animals that have both types of receptors. Pigs carry both α2,3 and α2,6 receptors and are thought to act as intermediate hosts in which avian and human viruses can reassort and generate viruses with the ability to overcome the host barrier (Ito et al., 1998). Recent evidence suggests that other animals, particularly certain terrestrial poultry, may also provide an environment similar to pigs by displaying both SAα2,3-gal and SAα2,6-gal receptors (Gambaryan et al., 2002; Wan and Perez, 2006). Antigenic drift (small mutations on the surface proteins within a single subtype) drives a particular subtype to evade the host immune response and can provide the capacity to infect other animal species. If a host is simultaneously infected with two influenza subtypes, mixing of the genetic material from the two virus strains can take place, creating a completely new strain. Antigenic shift can then occur when a naïve population is infected by a novel strain carrying a completely different HA subtype (although antigenic shift could result in the emergence of a new NA subtype, the NA surface protein seems to play a less important role than the HA in terms of significance in disease outbreaks or pandemics). The combination of antigenic drifts and shifts could produce a strain that could cause excess morbidity and mortality in susceptible hosts such as those that have caused pandemics in humans.
Influenza A viruses have plenty of opportunities to cross the species barrier. The establishment of a stable lineage and the spread thereof is, however, a rare event (Hinshaw et al., 1980a, b; Webster et al., 1978). The likelihood of a virus becoming endemic in a new host (recipient species) depends on two major factors: at the macro scale level, the interaction between donor and recipient animal species, and at the molecular level, the intricate interactions between the host and virus components (Kuiken et al., 2006). Influenza A viruses appear well adapted to the wild bird reservoir in which infections are rarely accompanied by signs of disease (Perkins and Swayne, 2001). Due to complex interactions of the virus in various wild bird species, the mechanisms involved in the genesis of novel influenza strains and the epidemiological factors implicated in the emergence of pandemic outbreaks are poorly understood. Some HA and NA subtypes appear more prevalent in certain aquatic bird species than in others (Kawaoka et al., 1988). In addition, these viruses appear to undergo cycles of prevalence that may last a few months to several years.
Avian influenza viruses may infect several domestic poultry species; the progression and outcome of the infection, however, varies within each species. For example, a particular isolate may produce severe disease in turkeys but not in chickens or any other poultry. The most virulent forms of highly pathogenic avian influenza (HPAI) are characterized by a highly fatal systemic infection that spreads to most organ systems including the cardiovascular and nervous systems (Acland et al., 1984; Gross et al., 1986; Slemons et al., 1991; Perkins and Swayne, 2002a, b, 2003). Morbidity and mortality can be as high as 100%, particularly in gallinaceous species. The incubation period is usually between 3 and 7 days depending on the virus isolate, age of the bird and bird species. Death may occur within 24 to 48 h after the onset of symptoms, but can be delayed for as long as one or two weeks. It is not the scope of this review to cover the clinical symptoms and histopathological aspects of the disease in different bird species, descriptions of which can be found in existing literature (Swayne and Suarez, 2000; Swayne and Halvorson, 2003). Avian influenza viruses from aquatic birds undergo significant selective pressure when adapting to chickens leading to definite changes in both the HA and NA genes (Matrosovich et al., 1999). The accumulation of basic amino acids at the cleavage site of the HA protein is a hallmark of the generation of avian influenza viruses with high pathogenic potential. Only AI viruses of the H5 and H7 subtypes have shown the potential to become highly pathogenic. An H5 or H7 virus can present itself as a low pathogenic virus but mutate without warning to become highly pathogenic, usually once they are introduced in domestic poultry, particularly chickens and turkeys (Kawaoka et al., 1984; Banks et al., 2001; Ito et al., 2001; Senne et al., 2006). Influenza subtypes other than H5 and H7 that can establish lineages in birds produce a mild disease, which can be exacerbated by secondary infections. For the HPAI viruses, the cleavability of the HA protein plays a critical role in the pathogenicity of AI viruses because it restricts tissue tropism (Kawaoka et al., 1987; Kawaoka and Webster, 1988; Walker and Kawaoka, 1993; Horimoto and Kawaoka, 1994, 1997; Rott et al., 1995; Steinhauer, 1999). HA is synthesized as a precursor, which requires post-translational cleavage by host proteases to create functional HA and produce infectious virus particles (Skehel and Wiley, 2000). All HPAI viruses identified to date differ from their low pathogenic avian influenza (LPAI) counterparts in the susceptibility of the HA to host proteases. HPAI are characterized by HAs that are highly susceptible to cleavage by numerous cellular proteases, which are ubiquitous in many cell compartments and organ systems. In contrast, the LPAI HA requires specific active extra-cellular proteases – such as trypsin – for cleavage and activation of infectivity. Trypsin-like proteases are restricted to the lumen of the respiratory and intestinal sites.
How do avian influenza viruses transfer from the wild bird reservoir into domestic poultry? One factor is agricultural and commercial practices that promote interspecies transmission and emergence of influenza viruses favoring the direct contact of natural hosts infected with influenza and non-natural hosts (Alexander, 1982). Live bird markets, backyard flocks, and free-range raised poultry are a few examples of such practices where natural and non-natural hosts of influenza can come in contact (Bulaga et al., 2003; Liu et al., 2003; Mullaney, 2003). These same conditions have also favored the transmission of avian influenza from poultry to humans.
The great variety of potentially susceptible domestic avian species increases the complexity and diversity of influenza strains in nature. Quail (Coturnix coturnix), for example, have been shown to provide an environment where influenza viruses from the wild bird reservoir can increase their host range and infect other avian species (Makarova et al., 2003; Perez et al., 2003a, b). Quail appear more susceptible than chickens (White Leghorn) to avian influenza viruses from wild birds, typically harboring asymptomatic infections (Makarova et al., 2003; Perez et al., 2003a). Quail usually take longer than chickens to show signs of disease from HPAI, while shedding substantial amounts of virus, increasing the chances for the spread of the virus (Webster et al., 2002). Interestingly, the respiratory tract of quail possesses a pattern of expression of sialic acid receptors similar to the one observed in humans; i.e. ciliated cells that express abundant SAa2,6Gal receptors and mucin-producing cells that express SAa2,3Gal receptors (Wan and Perez, 2006). Other studies suggest that pheasants could possess similar characteristics to those observed in quail (Humberd et al., 2006). Furthermore, domestic bird species that are infected with LPAI often do not show signs of disease during the period when the virus is effectively transmitted, whether in live bird markets, back yard flocks, or large and small commercial poultry operations. From an epidemiological perspective, however, the prevention and control of avian influenza in aquatic domestic poultry is the most crucial (Hulse-Post et al., 2005). Several studies have pointed to free-grazing ducks as major determinants in the spread of H5N1 among domestic poultry in South East Asia (Gilbert et al., 2006). Domestic aquatic poultry has most likely been responsible for the spill over of H5N1 into wild aquatic birds and the consequent massive geographic spread (Martin et al., 2006). There is no precedent for the presence of an HPAI virus that can infect so many wild bird species as well as mammalian species (cats, humans, mice and ferrets) and with such lethal outcome. Previous reports of HPAI in wild birds have been characterized by either no mortality or limited spread. The current H5N1 strains have undergone cycles of increased virulence and mortality for wild birds and domestic ducks and other aquatic birds, while maintaining a typical HPAI phenotype for terrestrial poultry. The potentially devastating ecological consequences of such events remain to be seen.
Before 1997, it was believed that molecular constraints restricted the host range of HPAI viruses to infection of avian species although limited cases of conjunctivitis were reported in humans (Campbell et al., 1970). Since the description of HPAI virus infection in 1878 as 'fowl plague', there has been little indication that the illness could be transmitted to humans, despite the multiple opportunities that have existed through the numerous outbreaks recorded in history. Since the 1990s, HPAI outbreaks have been reported frequently, caused by influenza viruses of subtype H5N1 in Southeast Asia (ongoing since 1997), H5N2 in Mexico (1994), Italy (1997) and Texas (2004), H7N1 in Italy (1999), H7N3 in Australia (1994), Pakistan (1994), Chile (2002) and Canada (2003), H7N4 in Australia (1997), and H7N7 in The Netherlands (2003) (Alexander, 2000; Munster et al., 2005). Some of these outbreaks have been associated with transmission to humans causing respiratory infections, usually in people in direct contact with infected poultry. Previous serologic studies in the United States and Europe provided evidence of human infections with various HA subtypes of avian influenza from poultry, however these infections were generally subclinical (Shortridge, 1992). The exceptions were H7 HPAI virus infections in The Netherlands and British Columbia, Canada that occasionally caused conjunctivitis in humans, but also resulted in one fatality in The Netherlands (Fouchier et al., 2004; Tweed et al., 2004). Human infections have also occurred with LPAI H9N2 strains that are endemic in Asia and have acquired receptor-binding properties similar to those present in human influenza viruses. However, it is H5N1 viruses that have infected humans more often and with more devastating outcome. The H5N1 virus that was first shown to give rise to strains that can transmit to humans appears to have emerged in Southeast China. During 1996, an AI outbreak was observed in geese in Guandong Province (Guo et al., 1998). Within a year, the goose virus reassorted, perhaps multiple times, with viruses circulating in other avian species (quail, chickens, teal). The new H5N1 virus was transported to Hong Kong, where it caused a major HPAI outbreak in chickens. The first bird-to-human transmission of this H5N1 virus was reported in a child that succumbed to the infection. Of the 18 human cases of H5N1 infection diagnosed at the time, six were fatal, prompting the rapid culling of approximately 1.5 million domestic birds in Hong Kong. This quick response likely succeeded in preventing additional human cases. Following the culling, the Hong Kong authorities adopted stringent prevention strategies that included the prohibition of aquatic birds in the live bird markets, monthly rest days in the poultry markets (during which lots were completely emptied and disinfected) and increased influenza surveillance in both wholesale and live bird markets. Unfortunately, similar strategies have not been implemented in other parts of Asia and, as a result, H5N1 viruses continue to circulate, particularly among geese and ducks, allowing genetic and antigenic evolution and increasing their geographic distribution and host range. Starting in 1999 in Hong Kong, avian species were monitored for virus through testing of blood and fecal samples and swabbing of cages. In May 2001, an avian H5N1 influenza virus was isolated from duck meat that had been imported to South Korea from China (Tumpey et al., 2002). Reports indicate that H5N1 circulated in Vietnam live bird markets during 2001 (Nguyen et al., 2005). From late 2002 through January 2003, mortalities of wild, exotic, and domestic birds due to HPAI were reported in Kowloon Park and other nature parks in Hong Kong (Ellis et al., 2004). In early February 2003, H5N1 viruses infected humans for the second time, resulting in two confirmed infections with one fatality (Peiris et al., 2004). Since late 2003, the world has experienced the largest outbreak of HPAI in birds in history. H5N1 outbreaks were reported in Vietnam, Thailand, Indonesia, Cambodia, Laos, Malaysia, China, Korea, and Japan. Massive die off of bar headed geese, among other wild bird species, were reported in May 2005 from Qinghai Lake, one of China's most beautiful natural reserves. Subsequently, Russia (Siberia), Kazakhstan, and Mongolia reported outbreaks of H5 in wild and domestic birds in late 2005 (Food and Agriculture Organization reports, www.fao.org). Despite culling and vaccination strategies the virus has spread even further. H5N1 outbreaks have been reported in Eastern Europe, the Middle East, and Africa in late 2005 through 2006, including Romania, Turkey, Greece, Croatia, Ukraine, and recently in Nigeria, Bulgaria, and Italy. Human infections have also occurred in Turkey and Iraq. A complete list of countries reporting outbreaks in wild and domestic birds and humans is presented in Fig. 2 and Table 1. The overall mortality rate of confirmed cases is approximately 50%, and the possibility of a pandemic remains as long as this virus continues to circulate in birds. With the possible role wild birds play in the epidemiology of HPAI, large regions of Asia, Middle East, Europe and Africa have become at risk. The HPAI H5N1 virus now poses a serious threat not only to Southeast Asia but also to the rest of the world.
Control of HPAI outbreaks have traditionally been resolved by the mass slaughtering of birds either infected or at high risk of becoming infected. This strategy must not be underestimated; it has been and will probably continue to be the most effective measure for the eradication of AI in infected flocks. However, when the viruses are widely spread in multiple domestic and wild avian species such as in the case of the Asian H5N1 epidemic, 'stamping out' alone is unlikely to be successful. Despite the approximately 150 million domestic poultry that have died or been culled in Asia to control the spread of the disease, the list of countries reporting outbreaks of H5N1 HPAI continues to increase. The estimated economic losses to the Asian poultry sector are around $10 billion, threatening the livelihood of millions of people (Perez et al., 2005). Thus other strategies to prevent and control the disease, such as vaccination, quarantine, and enhanced surveillance, have to be considered and used to complement stamping out policies.
Vaccination must be aimed primarily at reducing the presence of virus by reducing the number of susceptible animals or virus load. In Italy, vaccination of commercial poultry against H5 and H7 in high-risk areas combined with strict biosecurity measures and stamping out has resulted in the successful control of the spread of these viruses (Capua and Marangon, 2003; Capua et al., 2004; Marangon and Capua, 2006). However, no AI virus has been eradicated using vaccines alone. For example, continuous vaccination of chickens against a low-pathogenic H5N2 virus in Mexico over the last ten years has reduced disease signs and controlled the spread of the disease, however, the virus still circulates, particularly in the south of the country (Lee et al., 2004). The unnoticeable circulation of this virus may have contributed to its spread to neighboring countries including Guatemala and El Salvador. Influenza A vaccines in poultry include conventional inactivated oil-based whole AI virus, vectored virus, subunit protein and DNA vaccines. Currently, only inactivated whole AI virus vaccines and a fowl pox-vectored vaccine with an AI H5 HA gene insert are approved, prepared commercially, and used in various countries around the world (Suarez and Schultz-Cherry, 2000; Webster and Hulse, 2005). Current AI vaccines can provide a strong humoral response but typically do not produce a strong mucosal immune response and do not elicit adequate cellular immune response (Suarez and Schultz-Cherry, 2000; Wood et al., 2002; Capua and Marangon, 2004; Webster and Hulse, 2005). In regards to mass vaccination, current vaccines are associated with intensive labor requirements for parenteral administration, not to mention the cost of the vaccine, which is around 7 cents per dose.
Recent studies using live recombinant vectors have shown promising results with respect to efficacy as well as inexpensive methods for mass vaccination in poultry. In one of these studies it was shown that a live Newcastle disease virus (NDV) vector expressing the H5 gene from HPAI protected chickens from infection with NDV and HPAI (Veits et al., 2006). Likewise, an adenovirus-vector-based system expressing the H5 gene was shown to protect chickens against lethal challenge. An advantage of live vectors is that they are amenable to several vaccination strategies including aspersion and in ovo administration. A potential disadvantage of these systems is that prior exposure to the wild type agent limits the use of the vector. Studies are in progress to develop modified live attenuated avian influenza virus vaccines, which will have many of the benefits of live recombinant vectors and will not be affected by prior exposure to unrelated viruses or other agents.
Sporadic surveillance efforts have been key to the identification and characterization of potential epidemic and pandemic avian influenza strains. Since the late 1970's, live bird markets (LBMs) have been known to be a continuous source of influenza viruses (Shortridge, 1992). Nevertheless, the current H5N1 situation warrants a better understanding of the ecology, epidemiology, genetics and evolution of influenza viruses. This information has to be analyzed in the context of the ecology of their natural hosts, and of environmental, agricultural, and social risk factors that lead to the emergence of novel influenza strains. Rapid and adequate responses to contain outbreaks require coordinated and experienced animal influenza surveillance structures capable of deploying and implementing high throughput analysis and characterization of influenza viruses obtained from either animal species or humans. Such structure is either deficient or lacking in many underdeveloped countries. Maintaining a global animal influenza surveillance structure will undoubtedly lead to quick implementations for the control and eradication of outbreaks and to the prevention of human infections. A transparent global network animal surveillance structure will also allow rapid communication among participating countries with respect to outbreaks and whether outbreaks in more than one location are caused by a single virus strain or multiple related or unrelated strains. Intensive surveillance plans are being coordinated by the international community through organizations like the Food and Agriculture Organization (FAO), World Health Organization (WHO) and World Organization for Animal Health (OIE).
Since domestication, poultry has been considerably improved by natural and artificial selection. Modern selective breeding, particularly in chickens, has created significant progress in both egg and meat production (Romanov and Weigend, 2001; Burt, 2002). Current breeding strategies for commercial poultry focus on specialized production lines bred from large populations of a few breeds with genetic uniformity for the desired traits (Notter, 1999). A major issue with this breeding strategy is the erosion of local breeds and the loss of valuable genetic traits (Romanov, 1996). As an example, the selective process for increased meat production in broilers has resulted in breeds with increased incidence of congenital disorders, reduced fertility and reduced resistance to disease. Likewise, layers have experienced an increase in osteoporosis associated with rise in egg production (Burt, 2005). The search for disease-resistant genotypes or traits could be beneficial to a comprehensive disease control program for poultry production.
Like in mammals, innate immunity in birds is considered to be mediated by type I interferons (IFN). IFNs are responsible for the induction of genes with antiviral activity prior to the development of adequate T-cell and B-cell responses during primary infection. The Mx protein family is one group of proteins that have antiviral activity and are induced in response to IFN. The majority of animal species analyzed to date have one to three Mx protein isoforms with different antiviral activities and intracellular localization (Lee and Vidal, 2002). Mx proteins are small GTPase enzymes that appear to interfere with RNA transcription during virus replication (Staeheli, 1990; Nakayama et al., 1991, 1992; Samuel, 1991; Horisberger, 1992; Pitossi et al., 1993). In mice and rats, a nuclear Mx isoform (Mx1) blocks replication of influenza virus (Staeheli et al., 1986; Meier et al., 1990), whereas the cytoplasmic form (Mx2) inhibits vesicular somatitis virus (VSV) (Meier et al., 1990). The cytoplasmic human MxA protein shows antiviral activity in vitro against influenza, VSV, measles virus and Thogoto virus (Pavlovic et al., 1990; Haller, 1993; Schnorr et al., 1993; Horisberger, 1995). Birds, like mammals, produce Mx proteins, although their antiviral activity is less defined and remains controversial. The duck Mx, found in the nucleus and cytoplasm, does not appear to inhibit influenza replication, suggesting that the enhanced disease resistance to influenza in these birds is mediated by other unknown mechanisms (Bazzigher et al., 1993). Similarly, the chicken Mx protein, predominantly found in the cytoplasm, appears to have limited antiviral activity (Bernasconi et al., 1995). This protein is typically inactive in commercial breeds and attempts are being made to increase Mx expression or express the mouse Mx1 in chickens and evaluate its resistance to influenza and other viruses. Studies in transgenic pigs expressing the mouse Mx1 protein have yielded rather disappointing results in terms of enhanced anti-influenza activity (Muller et al., 1992).
AI viruses counteract the host's IFN response through the action of NS1, a small nuclear (∼20 kDa), dimeric protein that is highly expressed in cells early in the infectious cycle and has dsRNA-binding activity (Hatada and Fukuda, 1992; Nemeroff et al., 1995). The NS1 interferes indirectly with IFN through several mechanisms. One of them includes the sequestration of dsRNA thus preventing downstream events required for the activation of IFN-pathway genes such as IRF3, NFKB and AP1. The RNA-binding domain lies within the N-terminal 73 amino acids (Qian et al., 1995). The C-terminal remaining portion of NS1 is called the effector domain and includes binding sites for poly (A)-binding protein II (PABII) and the 30 kDa subunit of cleavage and polyadenylation specificity factor (CPSF) (reviewed in Krug et al., 2003). In addition, NS1 has been reported to interact with several other host factors, including the eukaryotic translation initiation factor elF4GI (Wolff et al., 1996, 1998; Falcon et al., 1999; Aragon et al., 2000; Burgui et al., 2003) and the protein kinase R (PKR) (Lu et al., 1995; Tan and Katze, 1998; Hatada et al., 1999). Activation of PKR in the presence of dsRNA (i.e. during virus infection) results in phosphorylation and inhibition of the translation initiation factor eIF2α subunit and thus inhibition of protein synthesis. The blocking of PKR activity by NS1 permits proper translation of viral products and progression of the infectious cycle. Strategies could be envisioned to inhibit influenza replication by controlling the synthesis of NS1. The exploitation of the RNA silencing pathway through small interfering RNAs (siRNAs) is an attractive alternative that could be applied to induce influenza resistance in birds. RNA silencing has been shown effective in vivo against influenza (Tompkins et al., 2004). If siRNAs are developed against multiple viral genes the chances of resistant variants can be greatly reduced. However, with a moving target like avian influenza, genetic-resistant approaches have to be considered with moderate optimism.
The unprecedented outbreak of H5 HPAI in Asia that has spilled over areas of Europe and Africa has brought a sense of urgency in terms of understanding the mechanisms that lead to interspecies transmission of influenza. The realization that domestic poultry species can act as a reservoir for the emergence of novel strains of influenza with pandemic potential requires important changes in the way domestic poultry are produced and traded. However, these changes also require major international efforts in order to safeguard public health and protect poultry while respecting traditions and cultures. A sustainable global surveillance network with transparent and open communications should act as a platform to prevent and control future outbreaks. Ideally, countries would develop and implement policies that encourage and reward the reporting of suspected outbreaks. The development of resistant traits in poultry and improvement of vaccines will only help if biosecurity and education programs are properly implemented. By focusing our efforts at the source, i.e. poultry species, the emergence of influenza strains with pandemic potential can be greatly reduced.
Request reprints from Daniel R. Perez
University of Maryland
Virginia-Maryland College of Veterinary Medicine
Department of Veterinary Medicine
8075 Greenmead Drive, College Park, MD 20742 (USA)
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Published online: August 02, 2007
Issue release date: July 2007
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