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Finding should aid those developing anti-virus vaccines. Vaccines have led to many of the world’s greatest public health triumphs, but many deadly viruses, such as HIV, still elude the best efforts of scientists to develop effective vaccines against them.

An improved understanding of how the immune system operates during a viral infection is critical to designing successful anti-virus vaccines. Scientists from the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health (NIH), have added an important dimension to this knowledge.

Focusing on mouse lymph nodes—bean-shaped organs that contain a variety of immune cells and are distributed throughout the body—the researchers discovered that immune cells confront viruses just inside of the lymph node and not deep within these organs as previously thought. The study, led by Jonathan Yewdell, M.D., Ph.D., chief of the NIAID Cellular Biology Section and his NIAID colleague, Heather Hickman, Ph.D., is described in a report online in Nature Immunology.

The results are significant, the authors say, as they observed in detail the interaction of viruses and immune cells inside a living organism, in this case, mice. Combining expertise from disciplines such as imaging, immunology, virology and other specialties, the scientists first extracted and then purified specific T cells—killer T cells—from mice. Killer T cells, which attack and kill infected or cancerous cells, are major
weapons in the immune system arsenal. The scientists labelled the T cells with a fluorescent marker, injected them back into the mice, and then infected the animals with vaccinia virus, the virus used to make smallpox vaccine, engineered to express a brilliantly coloured protein.

Using a multiphoton microscope, a highly specialized microscope that enables scientists to peer into a living
organism, the scientists could now look into the lymph nodes of the infected mice and see that the viruses had infected cells just inside the lymph node surface, triggering a swarm of T cells. These virus-specific T cells form an elaborate and dynamic communications network that activates them to divide and travel to the site of viral infection, where they kill virus-infected cells.

“A key challenge in viral vaccine research is developing strategies for immunizing against lethal viruses such as HIV that have eluded the standard vaccine approaches,” notes Dr. Yewdell. “We have contributed a page to the handbook of understanding how to rationally design vaccines to elicit a T-cell response.” According to the NIAID team, pinpointing where in the lymph node immune cells fight the virus should help efforts to design effective anti-virus vaccines.

The intriguing possibility that a virus which causes cancer in mice could also spread in humans has been raised by laboratory scientists……. again

The Austrian-led team found that mouse mammary tumour virus (MMTV) – which causes breast cancer in the animals – could replicate in human cells.

Other cancer experts, however, said the results, in the journal Retrovirology, should be treated with caution.

They said there was little evidence to link it to human breast cancer.

Viruses are now known to be involved in the development of several cancers – including cervical and liver cancer.

MMTV was discovered in the 1930s, and has been previously suggested as a possible cause of human breast cancer.

However, even though traces of the virus have been found before in human breast cancer cells, attempts to prove a link have foundered in the past because no-one could find evidence that the virus could survive and replicate in that environment.

The latest research claims to have done this – they say MMTV ‘rapidly spreads’ in breast cancer cells in their laboratory.
Dr Stanislave Indik, who led the team, said: “Often, viruses infect cells but cannot replicate further.
“If they can replicate, the chances that they cause disease may be increased.”

The researchers said that while not proving that the virus can cause breast cancer in real people, it “lends more weight” to theories linking the virus to the disease, and to other conditions such as the liver disease primary biliary cirrhosis.

They said that if the MMTV were to be proven, existing drugs such as the anti-HIV medication AZT would stop it replicating.

Other experts are no so convinced that MMTV is likely to be a culprit for the disease.

Epidemiologist Dr Rob Newton, from the charity Cancer Research UK, said: “This paper suggests that, under controlled laboratory conditions, a mouse virus can infect cultured cells derived from human breast tissue.
“It does not demonstrate that this actually happens in the real world, nor have the researchers shown that such infection leads to the development of cancer.

“At the present time, the overall evidence in this area does not support the view that MMTV is a cause of human breast cancer.”

This was echoed by Dr Sarah Cant, from Breakthrough Breast Cancer, who said: “Although this research indicates the mouse mammary tumour virus can spread between breast cancer cells in the lab, there is still no concrete scientific evidence that the virus causes breast cancer in humans.

“Much more research would be needed before we can say whether or not MMTV can be passed from mice to humans to cause breast cancer.”

The effort to find preserved samples of the 1918 influenza virus has been a pursuit of both historical and medical importance.

The influenza pandemic in 1918 was the most devastating single disease outbreak in modern history, and examining the virus that caused it may help prepare for, and possibly prevent, future pandemics. When the complete sequence of the 1918 virus was published in 2005, it represented a watershed event for influenza researchers worldwide.

An article in the journal Antiviral Therapy, scientists at the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health, narrate the story of how scientists discovered samples of the 1918 strain in fixed autopsy tissues and in the body of a woman buried in the Alaskan permafrost.

The article places this discovery in the context of decades of research into the cause of pandemic influenza, and the authors detail the strange convergence of events that allowed them to recover and sequence the virus in the first place. Its genetic material is so fragile that it should not have survived for days, let alone decades.

In a mass grave in a remote Inuit village near the town of Brevig Mission, a large Inuit woman lay buried under more than six feet of ice and dirt for more than 75 years. The permafrost plus the woman’s ample fat stores kept the virus in her lungs so well preserved that when a team of scientists exhumed her body in the late 1990s, they could recover enough viral RNA to sequence the 1918 strain in its whole entirety. This remarkable good fortune enabled these scientists to open a window onto a past pandemic. It could also help mankind gain a foothold for preventing a future one.

Here we go again. Another outbreak of gastroenteritis has affected 47 people at a nursing home at South Morang, North of Melbourne.

The illness struck 30 residents and 17 staff at the San Carlo Home for the Aged. Although it is still early, the source of the illness remains unknown.

One of the victims, a 90-year-old man, has died, but the cause of death is not yet known as it could be of natural causes.

San Carlo Home spokesman Christian Peterson says family members were asked to stay away from the home over the weekend.

“The best way people can help is by staying away and letting the staff get on with the job of looking after the residents and helping prevent the spread further,” he said.

“At all times we’ll keep relatives, staff and the appropriate agencies informed, as we have done over the last few days.”

A natural cleaning fluid made from bacteriophage could help meat processors get rid of pathogens from animal hides which is a key source of cross-contamination within processing plants.

The product became available on the market after the US Department of Agriculture gave approval for its bacteriophage treatment for killing E. coli O157:H7 on the hides of live animals must before they are slaughtered.

The bacteriophage product for E. coli can also be used to treat holding areas, transportation vehicles, containers and living quarters. USDA researchers also discovered that killing pathogens in hides before removal is an effective way of reducing the risk of carcass contamination.

Current treatment include broad spectrum antibiotics, harsh chemicals and irradiation have created super bugs, pollutants, harmed field workers and have even lowered the quality of some food products without effectively controlling the harmful bacteria.

Bacteriophages are specific against certain bacteria and it is believed that phages can succeed as a long-term solution for controlling unwanted bacteria where these old methods have failed. EDI Foods has already marketed LISTEX for lsiteria in cheese.

Biophage Pharma Inc has report its first financial result for 2006 – 2007. The company is a high-potential, revenue-driven biotechnology company focused on the development of an integrated approach for the prevention and control of bacterial infections within the food industry.

Overview of operations
Biophage reached an important milestone in its Biosensors Division in the first quarter of 2007 in the development of its compact PDS96 (R) Biosensor. The Corporation is now conducting extensive in-house performance testing and validation of this alpha prototype. Biophage also furthered the development of its four new biosensors: The BacTrapping(R) system, the micro-fluidics system, the “FastBac” biosensor and the e.sensor. In this context, Biophage signed an important collaborative agreement aimed at combining a2sp’s Magic Tag(R) immobilization technology with Biophage’s biosensor platform. Magic Tag(R) uses linkers, which are activated by daylight, for the immobilization of
biomolecules (including phages) onto different surfaces such as magnetic beads, biosensors and micro-array surfaces. Biophage and a2sp also jointly filed a patent application on February 16, 2007, relating to “methods for immobilizing viruses (phages) using photo-reactive linkers”.

In the Therapeutics Division, Biophage concluded initial sales of its LISTEX(TM) product to an important cheese producer in the U.S. Securing this
sale marked the beginning of a business relationship with our first client who intends to develop and use phage therapy as a biological solution to control potential Listeria monocytogenes (Listeria) contamination in cheese. On December 4, 2006, Biophage signed an MOU (memorandum of understanding) with EBI Food Safety (La Hague, Netherlands) for the sale and distribution of LISTEX(TM) in North America. LISTEX(TM) is the first bacteriophage product to receive FDA GRAS (Generally Recognized as Safe) recognition for the control of Listeria contamination in cheese.

In ImmunotoxLabs, Biophage hired Dr. Michel Heyne as director of its Beryllium Reference Lab to meet the increasing demand for Beryllium and MELISA(R) testing. With his vast experience in laboratory testing, this eminent hematologist will help expedite the accreditation process of
Biophage’s Beryllium laboratory by the Quebec National Institute of Public Health (INSPQ).

Financial Results
Contract revenues for the three months ended February 28, 2007 amounted to $107,551 compared with $195,944 in the same three month period in fiscal 2006. The decrease in substantially attributable to the completion of significant projects with important clients, although partially offset by an increase in revenues generated from Beryllium testing. Other income for the first quarter in 2007 reached $824 compared to $601 in the same period in fiscal 2006.

Research and development costs for the three months ended February 28, 2007 (before tax credits) amounted to $135,006, representing a $61,303, or 83% increase over the $73,703 recorded in the same interim period in the preceding fiscal year. The increase is substantially attributable to the hiring of additional staff affected to R&D, including a director for the Corporation’s Biosensors Division, commensurate with Biophage’s overall accelerated efforts in developing the phage therapy segment. Research and development tax credits for the first quarter amounted to $35,000, which compares to $20,000 for the three month period ended February 28, 2006, representing 26% and 27% of
related costs, respectively.

Costs of contracts for the three months ended February 28, 2007 amounted to $117,410, relatively unchanged from the $118,359 incurred during the same period in the preceding fiscal year. The slight decrease in the costs of contracts results from lower subcontracting and laboratory supply costs, which was almost entirely offset by an increase in salaries from the hiring of additional staff between the interim periods.

Biophage’s net loss for the three month period ended February 28, 2007 amounted to $261,287 ($0.01 per share) compared to a net loss of $157,821 ($0.00 per share) for the corresponding three month period in the preceding fiscal year.

Liquidity and Financial Resources
As at February 28, 2007 Biophage had cash and cash equivalents of $426,733 compared to $214,344 at November 30, 2006. The increase in cash and cash equivalents from November 30, 2006 levels is substantially attributable to the private placements completed during the interim period, although partially offset by cash used in operating activities (after changes in non-cash working capital items).

During December 2006 and February 2007, the Corporation issued 4,045,458 units through private placement. Each unit is made up of one common share and one common share purchase warrant, whereby each common share purchase warrant is exercisable for a period of two years at an exercise price of $0.17 per common share. The 4,045,458 shares were issued for a total cash consideration of $525,910. More detailed information regarding the foregoing can be found in the interim unaudited consolidated financial statements and related management
discussion and analysis which have been filed today on SEDAR at www.sedar.com.

Granting of Stock Options
On April 27, 2007, the Corporation’s Board of Directors granted stock options to purchase an aggregate 1,114,000 common shares of the Corporation at an exercise price of $0.12 per share to certain directors, employees and consultants of the Corporation, all of which vest immediately other than 150,000 options that will vest on the first anniversary of the grant and 150,000 options that will vest on the second anniversary of the grant. The grant of such stock options is made in accordance with the stock option plan of the Corporation. The granted options will expire on April 27, 2012.

About Biophage Pharma Inc.
Biophage Pharma is a high potential, revenue-driven Canadian biotechnology company focused on the development of innovative phage-based
products and technologies for the detection, prevention and control of bacterial infections. Founded in 1995, Biophage operates three divisions:

(1) The Biosensors division for the development and commercialization of Biosensors, more particularly a portable PDS96(R) Biosensor which is now in the pre-commercialization stage; (2) The phage therapy division for the prevention and control of bacterial contaminations in the medical, veterinary and environment fields; (3) The Immunotox Labs division, which provides services in Immunogenicity and Immunotoxicity, Beryllium sensitivity testing and MELISA(R) testing for the detection of sensitization to more than 200 different allergens including metals, penicillin, gluten and pollens.

Source: www.biophagepharma.net www.immunotoxlabs.com

Bird flu has again struck another victim in Egypt bringing the toll to 14. Marianna Kameel Mikhail who was 15 years old contracted the H5N1 bird flu virus last week.

She was admitted to hospital in Cairo on Thursday and died of respiratory failure on Tuesday evening. Although being treated in hospital with the antiviral Tamiflu, it was too late as she was admitted 7 – 10 days after developing advance symptoms. None of her family members were found to have the virus.

With a total of 34 humans who have caught bird flu in Egypt, 14 have died and 19 have recovered. A two-year-old girl from central Egypt is under treatment and is in good condition.

Egypt has the highest number of confirmed human bird flu cases outside Asia.

Did you know that bacteriophages which are commonly known as “phages” are naturally occurring viruses that infect and kill bacteria with very high specificity.

They do this by attaching to the surface of the bacterium, replicating inside, and eventually destroying their host. Most importantly, bacteriophages are specific for only one type of bacteria so the normal flora are left intact. Because replication is so rapid (one phage can produce 2 billion offspring in 2 hours) bacteria have little opportunity to develop resistance. In addition, once the target bacteria have been destroyed, the phages are no longer capable of reproduction, and subsequently disappear through natural processes, leaving no harmful residues. This environmentally friendly characteristic of phages is a major advantage when compared to conventional antibiotic use, where toxic residues have led to many problems. As such they represent a formidable, yet underutilised weapon in our constant war against bacterial infections.

Bacteriophage Therapy is the practical application of these very powerful lytic viruses to a bacterial infection, whether in animals, fish or even plants. The concept may appear novel but the fact is that it has been used for over 85 years in Eastern European countries like Georgia and Poland where it became part of the standard health care to treat burns, wound infections and gastrointestinal disorders.

Bacteriophages were discovered independently by two scientists between 1915 and 1917, more than 20 years prior to the isolation of Penicillin.

In 1915, Frederick Twort reported an “ultracosmic virus that somehow killed bacteria in solution”. Two years later Felix d’Hérelle a French-Canadian biologist identified and coined the name bacteriophage, meaning “bacteria eater”. Highly excited by the efficiency of the viruses against Shigella bacteria, d’Hérelle continued his studies on phages and was the first person to realise the potential of bacteriophages as therapeutic agents. In 1919 for the first time in history, he treated a 12 year old boy with severe dysentery; within 5 days of treatment the boy was completely cured.

After d’Herelle’s first successful use of phage therapy, other scientists around the world became interested in the new phenomenon and its potential as a therapeutic method.

Europe and the United States began to produce their own phages on a large scale for the medical treatment of cholera, typhoid fevers and bubonic plague. In 1932 in India alone, 191,000 vials of bacteriophages were used for the treatment of cholera. However, at the time, very little was known about phage biology and the very high specificity required for a successful treatment was poorly understood. Many famous scientists, including Bordet, disagreed violently with d’Herelle that the phenomenon was caused by a virus and argued that it was an enzyme or an active component present in the solution which caused the observed effect. Others thought that the solution simply stimulated the immune system, facilitating the healing process. Due to the resulting confusion doctors often used mismatched phages with a corresponding lack of success. Companies manufacturing the phages compounded the problem by making exaggerated claims and supplying poor quality products. It was hardly surprising that negative reports began to appear in the literature questioning the effectiveness of phage therapy.

By the late 1940’s, mired in controversy and with the widespread availability of penicillin and other broad spectrum antibiotics, phage therapy fell into decline and eventually vanished from the Western scientific radar, except as a bacteriology typing tool and as a platform for molecular biology. After all, the successful use of phage therapy depended heavily on the correct identification of a particular bacterial pathogen, plus the skills required for production. Why bother when the “magic bullet”, penicillin, came as a white powder, had broad spectrum of activity and was cheap and highly effective?

Fortunately, scientists in the former Soviet Union and various Eastern European countries, including Georgia and Poland persevered with their studies of phages. In Georgia, The Eliava Institute has been producing phages for the treatment of patients since 1930 and has recently attracted the attention of scientists anxious to benefit from their vast experience in the treatment of bacterial infections. However, despite the fact that phage therapy has been widely used and refined for over eighty years, it is still considered by Western science to be an experimental field because much of the development preceded the modern, more stringent regulatory standards of western pharmaceutical products.

A number of research institutions and companies around the world have taken up to the challenge of re-introducing bacteriophage therapy to our markets for the treatment of human, animal and agricultural bacterial infections. Initial studies have confirmed many of the original scientific observations. As a result, clinical trials have been set up in England, Germany and the United States to study the efficacy of phages. In the agricultural sector, the first phage based product for the use on tomatoes and pepper crops has received a commercial registration from the U.S Environmental Protection Agency (EPA). The FDA has also granted approval for a new core product of 6 lytic phages as a food additive in meat and poultry products to prevent infection with Listeria monocytogenes, a bacterial pathogen that affects more than 2500 people annually, especially pregnant women, newborns and immune-depressed individuals. In Wroclaw, the Institute of Immunology and Experimental Therapy (IITD) has been granted authorization to treat people with phages in cases where all else had failed.

The encouraging developments show that Phage Therapy has the potential to be a useful alternative antimicrobial therapy. However, much work still needs to be done to optimise the treatment protocols and to provide solid evidence on the safety of the treatment to the highest standards. Nonetheless, with the inevitable rise in antibiotic resistance and the diminishing pipeline of new antibiotics, phage therapy may prove to be a valuable and timely weapon in the fight against bacterial infections.

About the authors
Dr. Anthony Smithyman completed a PhD on Bacteriology and Immunology at Glasgow University in 1978. For the past 20 years he has managed Cellabs Pty, a Sydney-based diagnostics company, which specialises in Tropical and Infectious diseases.

Sandra Morales is a Microbiologist working for Special Phage Services Pty Ltd, Australia’s first phage-therapy company. She is also a PhD student in the University of Technology of Sydney and is currently undertaking an investigation of the potential use of bacteriophages in the treatment of antibiotic resistant infections. The project includes the screening of hundreds of Australian isolates against a broad collection of bacteriophages and studying their efficiency and potential for viable products.

References
EARSS Management Team, members of the Advisory Board, and national representatives of EARSS. (2006) EARSS Annual report 2005, On-going surveillance of S. pneumoniae, S. aureus, E. faecalis, E. faecium, E. coli, K. pneumoniae and P. aeruginosa. Bilthoven, Netherlands.

Häusler, T. (2006) Viruses vs Superbugs A solution to the antibiotic crisis? Houndmills, Basingstoke, Hampshire RG21 6XS and 175 Fifth avenue, Ney York, N.Y 10010: Macmillan.

Hayden, M.K. et al (2005) Development of Daptomycin Resistance In Vivo in Methicillin-Resistant Staphylococcus aureus. Clin Microbiol.; 43(10): 5285–5287

Kutter, E. & Sulakvelidze, A. (2005) Bacteriophages Biology and Applications. United States of America. CRC.

Noskin, G MD. et al. (2005) The Burden of Staphylococcus infections on Hospitals in the United States: An analysis of the 2000 and 2001 Nationwide Impatient Samples Database. Arch Inter Med, 165(1):1756-1761

Stone, R. (2002) Stalin’s forgotten cure The Forgotten Cure. Science, 298(5594): 728-31.

Sulakvelidze, A., Z, A. and G, M. (2001) Bacteriophage Therapy: Minireview. Antimicrob Agents Chemother, 45, 649-659.

Sulakvelidze, A. (2005) “Phage therapy: an attractive option for dealing with antibiotic-resistant bacterial infections.” Drug Discovery Today 10(12): 807-809.

Hepatitis A is a food borne virus that is transmitted via the faecal-oral route and infection is passed from person to person via foods and beverages contaminated with faeces or by direct personal contact.

Foods most susceptible include ready to eat foods that does not receive any further heat treatment. They include those eaten without cooking such as fresh produce which may be irrigated with contaminated water (salads), and bivalve shellfish (such as oysters) which may be grown in waters contaminated by human faecal effluent.

Oysters feed by filtering particulate matter such as algae from the surrounding water. They can accumulate viruses and other pathogenic microorganisms if they are present in the water during feeding. Once accumulated, viruses may take weeks to purge from an oyster, long after faecal coliform counts of the harvest waters have declined to permitted levels. The common practice of consuming oysters raw or mildly cooked means contaminating viruses within the oyster will not be inactivated prior to consumption, potentially resulting in food borne infection.

In Australia during 1997 there were more than 400 hepatitis A cases, including one death that was linked to consumption of contaminated oysters harvested from the Wallis Lakes area in NSW.

The source of the virus was probably untreated human sewage effluent which flowed from upstream of the oyster lease areas because of unusually high rainfalls. As a result of this outbreak, NSW adopted the internationally recognized Australian Shellfish Quality Assurance Program (ASQAP). The Program has been highly effective in the control of oyster-borne illness. In May 2006 Food Standards Australia New Zealand’s Primary Production and Processing Standard for Seafood was introduced. The Standard sets out food safety and suitability requirements for seafood generally from pre-harvesting production of the seafood up to, but not including manufacturing operations.

Research funded by the Australian Food Safety Centre of Excellence (AFSCoE) on the effects of High Pressure Processing (HPP) on hepatitis A virus in oysters was completed by a PhD student, Stephen Grove, in 2006.

The non-thermal technique of HPP is currently used in the United States to extend the refrigerated shelf life of oysters by inactivating spoilage microorganisms without altering sensory or nutritional qualities. Pressure is applied to oysters submerged in water within a pressure vessel and varies between 250 and 450 megapascal (MPa). Oysters are also shucked by the process, releasing the adductor muscles that hold the oyster shells tightly closed, enabling easy removal of meat without shell damage. One such company is Goose Point Oysters.

The AFSCoE PhD research investigated the effect of a range of pressure and processing times at different salt concentrations, on the viability of hepatitis A virus in artificially contaminated oyster meat. At a pressure of 450 MPa, a 100 fold reduction of hepatitis A in oyster meat was achieved by a 300 second treatment time. The sensitivity of hepatitis A virus to HPP was increased by lower salt concentrations within the oyster meat. A predictive model of the data was developed, and may be used by oyster processors to predict the inactivation of hepatitis A resulting from a particular treatment.

The development of the first tuberculosis vaccine is starting to move forward as clinical testing has begun on candidates. Tuberculosis is an airborne bacterial disease that kills nearly two million people annually. An estimated two billion, one of every three people on the planet are infected with the disease, although only ten percent of these people develop the disease and become contagious.

Novel subunit vaccine candidates aimed at boosting previous BCG-prime vaccination and novel viable attenuated vaccine candidates aimed at substituting BCG have both completed the preclinical stage.

Despite these achievements, rational vaccine design against tuberculosis has not come to an end. Novel findings in basic immunology and microbiology will advance further improvements in vaccine development. These include the potential role of cross priming to induce more potent T-cell responses, the role of memory T cells and regulatory T cells in sustaining or curtailing optimal immune responses, respectively, as well as the involvement of cytokines in T-cell migration to non-immunologic tissue sites and in the generation of memory. Knowledge about basic mechanisms underlying optimum protection will not only have a direct impact on future vaccine design against tuberculosis but also help in the formulation of a set of biomarkers with predictive value for vaccine efficacy assessment.

Baumann S et al, …
Curr Opin Immunol. 2006 Jun 12
Max Planck Institute for Infection Biology, Department of
Immunology, Schumannstrasse 21-22, 10117 Berlin, Germany.