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Creation of a cardiotropic adeno-associated virus: the story of viral directed evolution

Creation of a cardiotropic adeno-associated virus: the story of viral directed evolution | Virology News | Scoop.it

Adeno-associated virus (AAV) is an important vector system for human gene therapy. Although use of AAV serotypes can result in efficient myocardial gene transfer, improvements in the transduction efficiency and specificity are still required.  As a method for artificial modification and selection of gene function, directed evolution has been used for diverse applications in genetic engineering of enzymes and proteins. Since 2000, pioneering work has been performed on directed evolution of viral vectors. We further attempted to evolve the AAV using DNA shuffling and in vivo biopanning in a mouse model. An AAVM41 mutant was characterized, which was found to have improved transduction efficiency and specificity in myocardium, an attribute unknown for any natural AAV serotypes. This review focuses on the development of AAV vector for cardiac gene transfer, the history of directed evolution of viral vectors, and our creation of a cardiotropic AAV, which might have implications for the future design and application of viral vectors.

Ed Rybicki's insight:

A VERY interesting paper from the viewpoint of directed evolution of viruses AND from the standpoint of gene therapy.  Putting viruses to use!!

 

Adeno-associated virus particle courtesy of Russell Kightley Media

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Virology News
Topical news snippets about viruses that affect people.  And other things. Like zombies B-)
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Zika Virus: Two or Three Lineages?

Zika Virus: Two or Three Lineages? | Virology News | Scoop.it
Ed Rybicki's insight:
This is interesting for a number of reasons: one, because it nails down slightly more convincingly where Zika came from; two, because it introduces the concept of a wider range of genotypes than we knew about; three, because vaccines that might be expected to protect against Asian and African 1 types, might conceivably not protect against African II. And given the lesson of dengue types and vaccines and the potential for ADE, that might not be a good thing....
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Is Virology Dead? No - no, it's not!

Mark Twain once remarked that the reports of his death were greatly exaggerated. So too, the death of virology.

In certain quarters, it is now fashionable to declare the passing of virology. “Viruses are retro,” a faculty colleague once told me, deadly serious.

We have heard this before. In 1967, the U.S. Surgeon General allegedly proclaimed, “The time has come to close the book on infectious disease. We have basically wiped out infection in the United States” (1). This was before the arrival of AIDS and severe acute respiratory syndrome (SARS) and the discovery of hepatitis C virus, before the fear of an avian flu pandemic and bioterrorism.

Virology was once held in high esteem. In the first half of the 20th century, plant viruses held center stage. Studies of mosaic disease of tobacco revealed the existence of a new class of infectious agents smaller than bacteria, and tobacco mosaic virus taught us that viruses could be crystallized, disassembled, and reassembled into an infectious form: “life” could be studied with chemical approaches (2, 3). In the 1950s and 1960s, viruses that infect bacteria played a central role in the biological sciences. They formed the basis of the Hershey-Chase experiment, the first widely accepted evidence that DNA is the genetic material (4). Bacteriophage also led to the discovery of mRNA and the triplet nature of the genetic code and played a leading role in the birth of molecular biology (5). The 1970s and 1980s were a golden age for animal virology. The small genomes of many animal viruses and the ease of introducing them into cells made them the model organisms of choice to study eukaryotic cells. mRNA splicing, transcriptional enhancers, oncogenes, tumor suppressor proteins, antiapoptotic proteins, cellular trafficking signals and pathways, major histocompatibility complex (MHC) restriction, and much fundamental cell biology and biochemistry were discovered through studies of animal viruses (6). The roster of Nobel Prizes awarded for studies of viruses is long and unequaled.

The success of virology enabled the ascendancy of other fields. Restriction mapping, gene transfer into animal cells, directed mutagenesis, and whole-genome sequencing were developed to analyze small viral genomes (7–14). These powerful methods ushered in the recombinant DNA era and were in turn applied to studying cellular genes as well. In fact, much of genetic engineering, at least in the early days, centered on converting the much larger cellular genomes into virus-sized bits of genetic information, which could then be analyzed by the methods used so successfully on the viruses themselves. With the adoption of molecular cloning techniques by cell biologists and geneticists, virologists no longer had a monopoly on insights into the innermost workings of cells. Now that we can clone and study cellular genes and have sophisticated methods to analyze cells and whole organisms, so the argument goes, why settle for studying viruses?

To the cognoscenti, the real attraction of viruses was not only these methodological advantages but also the intimate relationship of viruses with their host cells. Because viruses depend on cellular machinery to replicate, they need to manipulate crucial regulatory nodes of cells to reprogram them into virus-producing factories (or into safe havens while waiting for the signal to replicate). By studying how viruses work the levers that control cell growth and behavior, and how cells fight back to maintain their sovereignty, important cellular processes are revealed. Thus, many aspects of signal transduction, cell cycle control, regulation of gene expression, immunology, and carcinogenesis were elucidated by studies of viruses and their interactions with host cells. Indeed, with their large population sizes, short generation times, and high rate of mutation, viruses are ideal evolutionary probes of cells. We may pride ourselves on the power of functional genomics screens, next-generation DNA sequencing, and sophisticated bioinformatics and proteomic analysis to dissect cellular activities, but these tools are no match for millions of years of fast-track viral evolution.
Ed Rybicki's insight:
Interesting article, albeit a couple of years old. Viruses are way too important for virology to be dead B-)
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Vaccination evokes gender-dependent protection against tularemia infection in C57BL/6Tac mice

Vaccination evokes gender-dependent protection against tularemia infection in C57BL/6Tac mice | Virology News | Scoop.it
Highlights

Gender bias is demonstrated in vaccination and protection against Ft challenge.

Following vaccination, female mice are better protected than male mice.

Ab, cytokine, and bacterial clearance studies are consistent with the above.

Gender bias is apparent with both Ft LVS and highly virulent Ft SchuS4.

Gender must be considered to accurately evaluate vaccine efficacy.
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Two Zika proteins responsible for microcephaly identified - possibly

Two Zika proteins responsible for microcephaly identified - possibly | Virology News | Scoop.it
USC researchers have tracked down two Zika proteins potentially responsible for thousands of microcephaly cases in Brazil and elsewhere -- taking one small step toward preventing Zika-infected mothers..
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One of the biggest vaccination drives ever is underway to beat yellow fever

One of the biggest vaccination drives ever is underway to beat yellow fever | Virology News | Scoop.it

What makes this vaccination campaign different or unique? This is one of the largest vaccination efforts to contain an ongoing outbreak ever undertaken. The response to the yellow fever outbreak in Angola, which started in December 2015, has already been remarkable. More than 10 million people have been vaccinated in Luanda Province and other affected areas of the country since February. The effect is already evident. No new cases in the area were recorded in July and the first weeks of August. But the outbreak has spread to the DRC. The current concern is for the evolving situation there as well as to prepare for possible flare ups of the disease in the coming rainy months. The vaccination campaign has been expanded to increase coverage in Angola in the areas that border the DRC in particular, and then in the DRC’s affected areas. These are regarded as high risk areas for the transmission of the virus.

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Development of a Zika vaccine

Currently, there is no approved vaccine against the Zika virus (ZIKV). However, several organizations are actively developing vaccines using various platforms and technologies. While many of these are in the early stages, several are based upon previously approved platforms and designs against dengue and other infectious disease agents. In March 2016, the World Health Organization (WHO) announced a list of organizations describing for approaches toward Zika vaccine development. This short editorial describes the overview of ZIKV and vaccine development.

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Smallpox could be about to return! Or - not, maybe?

Smallpox could be about to return! Or - not, maybe? | Virology News | Scoop.it
Smallpox – a deadly disease eradicated from the world in 1977 – could return as the frozen tundra of Siberia melts and releases the virus from the corpses of people who died in a major epidemic about 120 years ago, experts have warned.
Ed Rybicki's insight:
This is alarmism at its insidious best: shouting out a headline, based on flimsy evidence, that says "We're all going to die!" or similar nonsense.
Really: this IS nonsense.  Some corpses were found in the permafrost in Siberia, that MAY have had smallpox-like lesions on them, and from some of which which smallpox virus DNA could be recovered - presumably by PCR.
This does NOT constitute a threat of live virus being present, or escaping from the corpses even if it WERE there.
I can believe you could get live anthrax: those spores are incredibly tough, and can last for many years in soil, let alone in ice. I could also believe that one could find live megaviruses - the so-called pitho- and molliviruses - in permafrost, because their putative hosts are unicellular protozoans and because they are also seriously stable.
But smallpox? The virus is probably not as stable as the megaviruses mentioned; it relies for infection on its structure, which has membranes integral to it - AND it infects people, who, when they die, don't cool down very quickly, and whose cells release all sorts of nasty enzymes (lipases, proteases) as they die. Which could be expected to chew up most things, including poxviruses.
Oh, sure, poxviruses CAN survive for years at a pinch - in the form of dried secretions or scabs, which, because they are dehydrated and full of protein, tend to stabilise virus particles. This is how the old variolators and vaccinators (literally: people who used variola or "vaccine" to vaccinate against smallpox) used to preserve their inocula, when they weren't using fresh material.
Melting tundra is not like that, I will note: bodies with intact virions in them will thaw and rot all over again, and that rotting will reduce what little virus there may be even further.
So I am not a believer in Death From The Permafrost!
And nor should you be.
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Chikungunya infection: A potential re-emerging global threat

Infectious diseases are indeed a lifelong threat to everyone irrespective of age, sex, lifestyle and socio-economic status. The infectious diseases have persisted among the prominent causes of death globally. Recently, re-emergence of Chikungunya viral infection harmed many in Asian and African countries. Chikungunya was considered as a major threat in developing and under-developed countries; the recent epidemiological outbreak of Chikungunya in La Reunion urges the global researchers to develop effective vaccine against this viral disease. In this review, Chikungunya, pathogenesis and epidemiology were briefly described.

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It's not all Zika....
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A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea

A native promoter and inclusion of an intron is necessary for efficient expression of GFP or mRFP in Armillaria mellea | Virology News | Scoop.it
Armillaria mellea is a significant pathogen that causes Armillaria root disease on numerous hosts in forests, gardens and agricultural environments worldwide.
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Not a plant, but's not that far off...
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Researchers pinpoint key influenza-fighting immune trigger

Researchers pinpoint key influenza-fighting immune trigger | Virology News | Scoop.it
RT @InfluenzaTopNws: Researchers pinpoint key influenza-fighting immune trigger https://t.co/nHzZIcoDLF via @Aller_MD https://t.co/raHC3DlP…
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HPV vaccine can benefit older women

HPV vaccine can benefit older women | Virology News | Scoop.it
ALBUQUERQUE, N.M. — Journal Staff WriterA new study co-authored by a University of New Mexico researcher
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Virus Keychain

Virus Keychain | Virology News | Scoop.it
This clever and intricate keychain depicts a virus of a group know as the Herpesviridae. Every detail, right down to the capsid and envelope proteins has been rendered with great attention to accuracy! The lipid envelope opens up to reveal the iconic, icosahedral viral capsid, which in turn opens up to reveal the viral DNA. Even the unique types of capsomeres (capsid protein subunits) have been depicted. The Herpesviridae are a large group, which contains many important human pathogens, as well as viruses that infect a wide variety of other animals. Among the Human Herpes Viruses (HHVs).are Epstein Barr virus (the cause of mononucleosis), Varicella Zoster Virus (the cause of Chicken Pox), Cytomegalovirus, and the Herpes Simplex viruses. A unique, accessory sure to please virologists, medical professionals, or anyone who is biologically inclined! Dimensions (not including keyring or loop): 2.7 cm diameter Weight: 35.0 Grams
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Wa-a-a-a-nt!!!
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WHO calls for enhanced vigilance amidst reports of declining yellow fever cases

As the number of yellow fever cases decline in Angola and Democratic Republic of the Congo (DRC), the World Health Organization calls on Governments and partners to capitalize on the momentum and intensify response measures.
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History of Vaccines — A Vaccine History Project of The College of Physicians of Philadelphia

History of Vaccines — A Vaccine History Project of The College of Physicians of Philadelphia | Virology News | Scoop.it
The History of Vaccines explores the role of immunization in the human experience and examines its continuing contributions to public health.
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REALLY useful resource
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Dr. Donald A. Henderson, Who Helped End Smallpox, Dies at 87

Dr. Donald A. Henderson, Who Helped End Smallpox, Dies at 87 | Virology News | Scoop.it
Starting in 1966, Dr. Henderson, known as D.A., led the World Health Organization’s war on the smallpox virus, and achieved success astonishingly quickly.
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Great article on Henderson.
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Common cold viruses originated in camels - just like MERS 

Common cold viruses originated in camels - just like MERS  | Virology News | Scoop.it
There are four globally endemic human coronaviruses which, together with the better known rhinoviruses, are responsible for causing common colds. Usually, infections with these viruses are harmless to..
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Camel kissing obviously has a looooooong history...B-)
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Brazil Scientists Find Zika Traces in Different Mosquito Species

Brazil Scientists Find Zika Traces in Different Mosquito Species | Virology News | Scoop.it
Culex mosquitoes are more common and hardier than Aedes aegypti , which is known to transmit the virus
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Role of small biotechnology companies in biodefence vaccines

Role of small biotechnology companies in biodefence vaccines | Virology News | Scoop.it
Since Edward Jenner introduced immunization with cowpox in the late eighteenth century for smallpox prevention, vaccines have saved countless lives and trillions of dollars in public health and related expenditures. At the same time, a 40-billion-dollar-worldwide vaccine market has been created that is dominated by a few large pharmaceutical companies [1 Global vaccine market revenues in 2005, 2009 and 2015 (in billion U.S. dollars) [Internet]. 2015. [cited 2016 Apr 5]. Available from: http://www.statista.com/statistics/265102/revenues-in-the-global-vaccine-market/]. While the Food and Drug Administration (FDA) lists 80 licensed vaccine products [2 Complete List of Vaccines Licensed for Immunization and Distribution in the US [Internet]. Silver Spring (MD): U.S. Food and Drug Administration; 2015. [cited 2016 Apr 5]. Available from: http://www.fda.gov/BiologicsBloodVaccines/Vaccines/ApprovedProducts/ucm093833.htm], the number of diseases (22 pathogens or their toxic products) targeted is much smaller due to multiple competing products for high-value markets. This is a sobering reminder that successful vaccine development is a colossal undertaking plagued with risks and requires companies with a strong financial backbone as well as extensive experience and infrastructure.
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Deadly African virus 'on brink of spreading to Europe and Americas'

Deadly African virus 'on brink of spreading to Europe and Americas' | Virology News | Scoop.it
A deadly African virus is on the brink of spreading to Europe and the Americas amid the largest outbreak in more than 30 years, a charity has warned. Yellow fever can cause bleeding from the ears, eyes and nose, organ failure, jaundice and death in the most severe cases, and is considered such a threat that many African nations refuse entry to anyone who has not been vaccinated. Yet despite those regulations, thousands of suspected cases have been reported in the Democratic Republic of Congo (DRC) after the disease crossed the border from Angola. 
Ed Rybicki's insight:
"Yellow fever can cause bleeding from the ears, eyes and nose, organ failure, jaundice and death in the most severe cases, and is considered such a threat that many African nations refuse entry to anyone who has not been vaccinated."

And you folk worry about Zika?
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Olympic dream crushed by penis

Olympic dream crushed by penis | Virology News | Scoop.it
NEVER before has a member of the male species wished for a smaller manhood. Until now.
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Just. Couldn't. Resist...B-)
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Could SMALLPOX return from the grave?

Could SMALLPOX return from the grave? | Virology News | Scoop.it
Scientists are worried that the deadly disease smallpox could return because permafrost is melting close to where hundreds of infected bodies were buried in Siberia, Russia.

Via Ian M Mackay, PhD, Chris Upton + helpers
Ed Rybicki's insight:
Ya. No. I am willing to bet the possibility of there being any viable virus there is so small as to be no chance at all.  But they should test it.
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Viruses 'more dangerous in the morning'

Viruses 'more dangerous in the morning' | Virology News | Scoop.it
Viruses are more dangerous when they infect their victims in the morning, a University of Cambridge study suggests.
The findings, published in PNAS, showed viruses were 10 times more successful if the infection started in the morning.
And the animal studies found that a disrupted body clock - caused by shift-work or jet lag - was always vulnerable to infection.
The researchers say the findings could lead to new ways of stopping pandemics.
Viruses - unlike bacteria or parasites - are completely dependent on hijacking the machinery inside cells in order to replicate.
But those cells change dramatically as part of a 24-hour pattern known as the body clock.
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New Tamiflu study suggests drug does reduce flu impact considerably

New Tamiflu study suggests drug does reduce flu impact considerably | Virology News | Scoop.it
A new analysis of research regarding oseltamivir - marketed and commonly referred to as Tamiflu - has found that the drug reduces the duration of flu symptoms and the risk of respiratory tract infections, according to its authors.
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Polio eradication faces setback as Nigeria records first cases in two years

Polio eradication faces setback as Nigeria records first cases in two years | Virology News | Scoop.it
Health officials launch emergency immunization campaign.
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POLIOMYELITIS UPDATE - NIGERIA

POLIOMYELITIS UPDATE (11): NIGERIA (BORNO)
******************************************
A ProMED-mail post
http://www.promedmail.org
ProMED-mail is a program of the
International Society for Infectious Diseases
http://www.isid.org

In this update:
[1] EpiCore member responses to RFI on this outbreak
[2] Advanced notice by Polio Eradication Initiative 11 Aug 2016

******
[1] EpiCore member responses to RFI on this outbreak
Date: Thu 11 Aug 2016
Source: EpiCore Global Surveillance Project [edited]

[Below is a summary of some of the very helpful responses received today (11 Aug 2016) to an RFI submitted through the EpiCore Surveillance Project. We are very grateful to the EpiCore Project members for their rapid responses (and in some cases submitting follow-up responses as more information became available). Of note, in less than 12 hours we have received 17 responses and they are continuing to come in. - Mod.MPP]

a - Both cases are WPV1. Each in Jere and Gwoza local government areas.

b - The virus is type 1, all the children are under 3 years of age and have received more than 5 doses. The state has been asked to prepare an immediate response plan to cover the 2 affected wards in Jere and Gwoza LGAs. bOPV is available for this and subsequent rounds that will be mounted in wider areas, targeting more children than the immediate response round. More details will come your way soon. Gwoza LGA because of the insurgents there, is not fully accessible. [Posted already in Poliomyelitis update (10): Nigeria (BO), new cases, RFI 20160811.4408853 earlier today]

c - There are 2 cases of WPV in Gwoza and Maiduguri in Borno State, Nigeria

d - Confirmed. Circulating indigenous strain. Hard to reach area of Borno State. Poor surveillance due to insecurity.

e - WPV1 has been confirmed. Yes there are 2 polio cases.

f - WP1 have been confirmed in Borno State, Nigeria. The contact of an index case is positive for WP1 while the index case is negative --Jere Local Government area (LGA) in Borno State. Another index case and 3 contacts have WP1 -- Gwoza LGA. Note: The affected LGAs are security challenged areas
From: Professor Marycelin Baba

g - Follow-up information from Professor Marycelin Baba on outbreak of WP1 in Borno State, Nigeria:
- The case from Jere LGA
A 3 year old male, vaccination : 5 doses (last dose 12 Jul 2016)
- Index case from Gwoza
A 2 year old female, vaccination : 6 doses (last dose 27 Jul 2016)

- Contacts
No.1 contact: 4 year old male, vaccination: 8 doses (last dose 20 Jul 2016)
No.2 contact: 3 year old male, vaccination : 6 doses (last dose 19 Jul 2016)
No.3 contact: 5 year old female, vaccination : 10 doses (last dose 20 Jul 2016)

h - Gwoza LGA - One child with acute flaccid paralysis, onset of paralysis on 13 Jul 2016 and close healthy contacts of child. Stool specimen collected on 21 Jul 2016
Jere LGA - A healthy contact of child who developed AFP on 6 Jul 2016 (index case has a negative test result. Stool specimen collected on 22 Jul 2016).
Genetic sequencing suggests these isolates are mostly linked to WPV1 last detected in Borno in 2011

i - Two cases of WPV 1.
No.1 case: Location Jere LGA of Borno State; Ward: Dusuman Ward; date of onset: 4 Jul 2016
From a child at Muna IDP camp housing IDPs from Marte, Mafa, Jere and Dikwa LGAs, which have been inaccessible for over two years

No 2 case: Location Gwoza LGA of Borno State; Ward: Hausari; date of onset: 5 May 2016

WPV 1 an orphan virus circulating since 2011

Gwoza LGA fell to insurgents on the 5 August 2014 it has 13 political wards as at today 5 ward headquarters are accessible

Actions:
- immediate investigation ongoing
- immediate Vaccination to commence on 15 Aug 2016
- more detail plans for 3 rounds of immunization ongoing
- AFP external peer review called off
More updates will be made available as they unfold

j - Nigeria, wild polio virus type one (WPV1) has been detected from Borno state. Virus was isolated from 2 local government areas (LGA) of Borno; in Gwoza LGAs, in a child with acute flaccid paralysis (AFP - onset of paralysis on 13 Jul 2016) and close healthy contacts of that child; and in Jere from a close healthy contact of a child who had developed AFP symptoms on 6 Jul 2016.

It is the 1st WPV1 detected in Nigeria since July 2014. Genetic sequencing of the isolated viruses suggest they are most closely linked to WPV1 last detected in Borno in 2011, indicating the strain has been circulating without detection since that time.

An outbreak response plan is currently being finalized, consisting of 3 large-scale supplementary immunization activities (SIAs) with bivalent oral polio vaccine (OPV), the 1st one beginning within 2 weeks, and subsequent rounds being conducted at short intervals (of between 2 to 3 weeks).
Ed Rybicki's insight:
Very sad this is happening.
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