Pfizer and Protalix to Develop and Commercialize Treatment for Gaucher’s Disease 
 

Target Health congratulates its client Protalix for this excellent opportunity.
 

Target Health was Protalix’s CRO partner and together with Cato Israel, who monitored outside of North America, we took this product from toxicology to the pre-IND meeting, IND submission, Phase I, Phase III, Orphan Drug, Fast Track, Expanded Access, multiple FDA meetings and now the eCTD NDA submission. Target e*CRF® and Target Document® were used for the pivotal trial and is now being used in four additional protocols. This development program was the best case of a Sponsor/CRO/FDA partnership. The pre-IND meeting took place in June 2004, and last patient last visit was September 2009.

Target Health’s team was lead by Glen Park PharmD. Dr. Park joined Target Health in 2005 and since then has been directly involved with approvals of 1 NDA (head lice), 1 MAA (emergency contraception) and 1 PMA (adhesion prevention in the newborn). There are currently 2 NDA submissions in review (emergency contraception and cystic fibrosis) and quite a few programs in the hopper.

Press Release

Pfizer Inc. and Protalix, Ltd. today announced that they have entered into an agreement to develop and commercialize taliglucerase alfa, a plant-cell expressed form of glucocerebrosidase (GCD) in development for the potential treatment of Gaucher’s disease. Under the terms of the agreement, Pfizer will receive exclusive worldwide licensing rights for the commercialization of taliglucerase alfa, while Protalix will retain the exclusive commercialization rights in Israel. Taliglucerase alfa is the first enzyme replacement therapy derived from a proprietary plant cell-based expression platform using genetically engineered carrot cells.

With the successful completion of Phase III clinical studies, Protalix is preparing to complete a rolling NDA with the FDA. The FDA has granted Orphan Drug designation and Fast Track status, facilitating the development and expediting the review of drugs to treat rare conditions or diseases, as well as an Emergency Use Authorization. The FDA has also requested, and subsequently approved, an Expanded Access Program (EAP) treatment protocol. Taliglucerase alfa is currently being provided to Gaucher’s patients in the U.S. under the EAP protocol, as well as to patients in the European Union under a compassionate use protocol.

The-Scientist.com, December 2, 2009, by Ed Yong  —  If I say the phrases ‘anti-ageing’ and ‘nutritional balance’ to you, you’d probably think of the pages of quack websites selling untested supplements than the pages of Nature. And yet this week’s issue has a study that actually looks at these issues with scientific rigour. It shows that, at least for fruit flies, eating a diet with just the right balance of nutrients can lengthen life without the pesky drawback of producing fewer offspring.

20091203-13

Despite the claims of the cosmetic and nutritional industries, chemicals or techniques that slow the ageing process are few and far between. We’re a long way from any fountains of youth, but there is at least one conclusive way of extending an animal’s life – restricting the calories it eats. It works in yeast, flies, worms, fish, mice, dogs and possibly even primates, but it comes at a cost. The dieting organisms had lower reproductive rates (technically, they had lower ‘fecundity’).

Scientists suspected that eating fewer calories mimicked the effects of famine and food shortages. In such conditions, parents who breed put their health at risk and their offspring’s odds of survival are slim anyway. So animals divert their resources to maintaining their health at the cost of their fecundity. This explanation suggests that survival and reproductive success are at odds with one another – fewer offspring is simply the price of a longer existence.

But Richard Grandison and Matthew Piper have found that this isn’t true. Working with Linda Partridge at University College London, they have shown that you can improve both the fecundity and lifespan of a fruitfly by supplementing its restricted diet with the amino acid methionine. The trick won’t work in exactly the same way for other animals so don’t go bulk-ordering methionine yet. However, the results do prove the point that flies can have their cake (or lack thereof) and eat it, provided the cake has just the right balance of nutrients.

Grandison and Piper fed Drosophila flies with diluted stocks of yeast, so they are the same amount that they would normally do, but had fewer calories to show for it. As usual, their lives increased and their reproductive rates fell. The duo tweaked the diet until it gave the flies the maximum possible lifespan and then systematically added back nutrients until they hit on some that would restore their fecundity while retaining their extra years.

Vitamins didn’t do it; nor did fats or carbohydrates. Extra doses of essential amino acids improved fecundity but it brought lifespan down too, as if the flies had eaten a full meal in the first place. This shows that calorie-restricted diets do their thing because they change the levels and ratios of amino acids in a fly’s food.

Grandison and Piper discovered that one particular amino acid, methionine, was crucially important. Methionine is a boon to reproduction, but it conspires with other amino acids to shorten lifespan. Without methionine, the flies lived to a ripe, old age but their fecundity faltered. The best combo was methionine on its own, without the other amino acids – that boosted fecundity and maintained the flies’ extended lifespans.

20091203-14

These results clearly show that survival and reproduction aren’t opposed – you just have to get the right balance of nutrients. Getting that balance could be the key to achieving the same winning combo of longer life and better reproductive success without actually cutting down on the calories.

But clearly, there’s a massive word of warning to all of this: methionine happens to be the magic ingredient for flies fed on yeast. Going out and buying methionine supplements is not going to turn you into an immortal Casanova. In this study, methionine only worked in a restricted diet where other amino acids are scarce. Likewise, in previous studies, mice and rats live longer if they cut down on methionine.

The main message from this study is that lifespan and fecundity don’t always trade-off against one another – getting the ideal balance of nutrients unlocks the best of both worlds. It’s likely that the same principle applies to other animals, because the biology of ageing is remarkably consistent across species, but we still don’t know where the point of balance rests. Look on the shelves of a health store and you might think that we’ve got questions like this cracked. We don’t – ageing research is in its infancy and there’s a lot of work left to be done.

Reference: Nature doi:10.1038/nature08619

 
Secrets of the supercentenarians: Life begins at 100
The-Scientist.com, by Ed Yong  —  People aged 100 or over, are one of the fastest rising demographics in the world and their numbers will surely swell even further with ageing populations and advances in modern medicine. The feature looks at what happens when people reach these extreme ages and what happens to them when they do.

It ended up being surprisingly optimistic. Far from being a helpless drain on society, there’s growing evidence that a substantial proportion of centenarians lead fulfilling and independent lives. Indeed, a study involving everyone in Denmark born in 1905, found that the loss of independence that comes with age is balanced out by the fact that the sickest people die earlier. The upshot is that the proportion of people who can take care of themselves remains steady and extreme longevity doesn’t lead to extreme disability.

What happens to the two sexes in extreme old age, is that women are more likely to get there but the men who do, tend to be fitter.  The diseases that affect the oldest old – cancer, chronic diseases and Alzheimer’s are rare, but other forms of dementia and arthritis are common. Researchers are obtaining a growing knowledge of the “centenarian genome” and what it tells us about the ageing process.

THIS year, the number of pensioners in the UK exceeded the number of minors for the first time in history. That’s remarkable in its own right, but the real “population explosion” has been among the oldest of the old – the centenarians. In fact, this is the fastest-growing demographic in much of the developed world. In the UK, their numbers have increased by a factor of 60 since the early 20th century. And their ranks are set to swell even further, thanks to the ageing baby-boomer generation: by 2030 there will be about a million worldwide.

These trends raise social, ethical and economic dilemmas. Are medical advances artificially prolonging life with little regard for the quality of that life? Old age brings an increased risk of chronic disease, disability and dementia, and if growing numbers of elderly people become dependent on state or familial support, society faces skyrocketing costs and commitments. This is the dark cloud outside the silver lining of increasing longevity. Yet researchers who study the oldest old have made a surprising discovery that presents a less bleak vision of the future than many anticipate.

It is becoming clear that people who break through the 90-plus barrier represent a physical elite, markedly different from the elderly who typically die younger than them. Far from gaining a longer burden of disability, their extra years are often healthy ones. They have a remarkable ability to live through, delay or entirely escape a host of diseases that kill off most of their peers. Supercentenarians – people aged 110 or over – are even better examples of ageing gracefully. “As a demographic group, they basically didn’t exist in the 1970s or 80s,” says Craig Willcox of the Okinawa Centenarian Study in Japan. “They have some sort of genetic booster rocket and they seem to be functioning better for longer periods of time than centenarians.” The average supercentenarian had freely gone about their daily life until the age of 105 or so, some five to 10 years longer even than centenarians, who are themselves the physical equivalent of people eight to 10 years their junior. This isn’t just good news for the oldest old and for society in general; it also provides clues about how more of us might achieve a long and healthy old age.

One of the most comprehensive studies comes from Denmark. In 1998, Kaare Christensen at the University of Southern Denmark, in Odense, exploited the country’s exemplary registries to contact every single one of the 3600 people born in 1905 who was still alive. Assessing their health over the subsequent decade, he found that the proportion of people who managed to remain independent throughout was constantly around one-third of the total: each individual risked becoming more infirm, but the unhealthiest ones passed away at earlier ages, leaving the strongest behind. In 2005, only 166 of the people in Christensen’s sample were alive, but one-third of those were still entirely self-sufficient (Proceedings of the National Academy of Sciences, vol 105, p 13274). This is good news from both personal and societal perspectives, for it means that exceptional longevity does not necessarily lead to exceptional levels of disability.

Christensen’s optimistic findings are echoed in studies all over the world. In the US, almost all of the 700-plus people recruited to the New England Centenarian Study (NECS) since it began in 1994 had lived independently until the age of 90, and 40 per cent of supercentenarians in the study could still look after themselves. In the UK, Carol Brayne at the University of Cambridge studied 958 people aged over 90 and found that only one-quarter of them were living in institutions or nursing homes. Likewise, research in China reveals that before their deaths, centenarians and nonagenarians spend fewer days ill and bedridden than younger elderly groups, though the end comes quickly when it finally comes.

Doubting your diagnosis? Read on to find out what you might really have.

 

WebMD.com, December 2, 2009, by Laura Nathan
Doubting your diagnosis? Read on to find out what you might really have.

Sometimes even the best doctors miss the mark: About 40 percent of all mistakes that M.D.s make are misdiagnoses, says the National Patient Safety Foundation. That’s because many ailments have similar symptoms or can be detected only with tests that your physician might consider unnecessary if he’s confident in his verdict. If you’re in the know about often-confused conditions, though, you can ask the right questions to prevent or fix an error – and even save your life.

1. YOUR SYMPTOMS: Numbness on one side, headaches, dizziness, suddenly blurred vision, lack of balance or muscle coordination, and/or slurred speech

  • The doctor says it’s: Vertigo, migraines, or an inner-ear disorder
  • It could be: Stroke
  • Why the confusion? Research shows that 14 percent of stroke cases in people under 45 are misdiagnosed. When patients are young and otherwise healthy, ER staff might point to milder problems first. But if you leave the ER with an undiagnosed stroke, you could suffer another one. You may also miss the chance to reverse impaired speech or vision, paralysis, and brain damage.
  • Red flags: If one side is numb or you have any combination of the listed symptoms, rush to the ER, especially if symptoms persist more than an hour.

2. YOUR SYMPTOMS: Headaches and/or ringing or aching ears, plus aching back, neck, and/or teeth

  • The doctor says it’s: Migraines or an ear disorder
  • It could be: Temporomandibular joint syndrome
  • Why the confusion? When the joint connecting your jaw and skull becomes inflamed, the pain radiates and causes headaches or ear problems. TMJ is best treated by a dentist, but the symptoms will likely send you to an M.D., who might diagnose you with something else. The price you’ll pay? Serious pain that could easily have been alleviated.
  • Red flags: If your doctor gives a tentative diagnosis without conducting any tests or prescribes meds that don’t do the trick, head to a dentist.

3. YOUR SYMPTOMS: Fatigue or trouble breathing plus chest pain or tightness and/or palpitations

  • The doctor says it’s: Stress or panic attack
  • It could be: Heart attack or heart disease
  • Why the confusion? Heart attacks tend to be more subtle in women than in men: Fatigue or shortness of breath might be your only sign of a problem. In fact, up to half of female heart attack victims are initially misdiagnosed – and heart disease remains the top killer of women in America.
  • Red flags: If your doc’s recommended treatment doesn’t help, see a cardiologist, pronto.

4. THE SYMPTOMS: Sadness plus fatigue, weight gain, insomnia, and/or muscle aches or stiffness

  • The doctor says it’s: Depression
  • It could be: Hypothyroidism
  • Why the confusion? Doctors tend to link persistent sadness with depression and might not think to test for hypothyroidism, a condition in which your thyroid gland fails to produce enough hormones. Left untreated, hypothyroidism can cause high cholesterol, high blood pressure, heart disease, and (ironically) clinical depression.

Red flags: If the meds your doctor prescribes don’t lift your mood, request a TSH blood test to check for hypothyroidism. Better yet, ask for the test during your initial visit.

The-Scientist.com, December 2, 2009  —  It’s been a tough year for every industry, and the life sciences are no exception. Yet companies and academic laboratories across the globe have developed innumerable new products designed to take your research to the next level. But with many lab budgets tighter than last year, which technologies are worth the investment?

That’s why, for the second year in a row, we have gathered a panel of expert judges to pick the year’s best innovations to hit the life sciences market in the past year. This year’s winners run the gamut from imaging, genomics, and other tools that stunningly capture both intracellular and extracellular processes. Our judges-Steven Wiley, Jean Wang, Shawn Levy, and David Piston-are all known for pushing the technical boundaries, and have collectively published more than 600 academic papers.

It may have been a tough year for industry in general, but it was a great one for life science innovation.

 

Cell culture in 3D

20091203-3

The Benchtop BioLevitator, which combines an incubator and a centrifuge into a single unit, is one of the first 3D cell culture systems.

“This is a completely new kind of technology,” says Amy Schneck, assistant product manager of the Hamilton Company, which developed the instrument. Besides creating a 3D culture, which is closer to an in vivo environment, the BioLevitator also allows researchers to grow more cells in less time relative to 2D culture, Schneck adds. Global Cell Solutions, a partner company, developed a unique microcarrier-a matrix lined with proteins-that facilitates cell growth on the 3D surface.

The BioLevitator can grow four cell culture tubes at once and also contains internal magnets that keep cells suspended and homogenous. Multiple protein coatings support different cell lines. During the culture, each tube is monitored for carbon dioxide, temperature, cell density, and pH. When cultures are complete, all data can be transferred to a computer for analysis using the BioLevitator’s USB port.

At $35,000, this compact, multipurposed instrument is also environmentally friendly because it works more efficiently than 2D systems, reducing the use of harsh chemicals and labware required for other instruments. As a result, Hamilton estimates that the 3D system can cut annual costs by 60 percent when culturing 40 million Chinese hamster ovary cells per week. The BioLevitator will be available in December 2009.

LEVY: The benchtop size and microprocessor-controlled and -monitored environment, coupled with innovative use of magnetic fields to maintain cells in suspension, makes the BioLevitator an innovative product in a very traditional field.

WILEY: This is a compact unit to greatly simplify microcarrier-based cell culture, which is usually a very complex system to implement. This should allow high-density culturing of anchorage-dependent cell lines, which are usually more physiologically relevant than anchorage-independent ones.

 

New recipe for protein expression

20091203-4

Synthetic genes are considered the most cost-efficient, timely, and flexible tool for achieving high levels of protein expression, a fundamental component of modern biotechnology research. But since different codons can produce the same amino acid, scientists have innumerable combinations to choose from when encoding a protein. And some combinations produce better results than others. Typically, researchers use anecdotal evidence to pick which set of codons will optimize protein expression, with hit-or-miss results. Now, scientists from the California-based company DNA2.0 have developed new design algorithms to predict the best set of codons to use based on actual gene characteristics. The system, described in the September issue of PLoS ONE, (4(9): e7002), produces protein expression up to 10 times better than previous approaches, says Mark Welch, the director of gene design for DNA2.0.

The team designed, synthesized, and expressed varied sets of genes encoding two different proteins (a DNA polymerase and a single-chain antibody) and, based on which codons produced the most protein, developed a design principle to predict the gene combinations that optimize protein expression.

The company made their E. coli algorithm free when they published it in PLoS ONE, but their yeast algorithm will cost up to $25,000 per year for use on an infinite number of genes, says Claes Gustafsson, the company’s vice president of sales and marketing. The price of already-made algorithms for other species varies depending on the size of the requesting institution and number of genes that need to be synthesized. The company can also develop algorithms for new hosts from scratch, but the process can take up to a year and cost between $100,000 and $250,000. The technology is still so new, Gustafsson says, that “the exact business plan is still up in the air.”

PISTON: This is another important milestone towards the use of fully synthetic genes, especially for protein engineering applications.

WILEY: Very nice! Definitely innovative thinking going on here.

 

New measure of metabolism

20091203-5

Invented by Seahorse Bioscience in Massachusetts, the XF96 Analyzer is the first instrument that can measure the two major energy pathways in cells-mitochondrial respiration and glycolysis-providing a comprehensive picture of cellular metabolism and how that process goes awry in disease. “Before this instrument, we could never do the magnitude or complexity of experiments,” says Steve Chomicz, vice president of sales & marketing at Seahorse Bioscience.

Prior to the XF96 Extracellular Analyzer, scientists relied on the Clark electrode technology for measuring cellular oxygen consumption, a time-consuming technique that provided minimal information. Now in just 35 to 90 minutes, the XF Analyzer can measure oxygen consumption-an indicator of mitochondrial respiration-as well as extracellular acidification, which is a byproduct of glycolysis. After isolating a small volume of cells in a microplate, the instrument can measure the change in dissolved oxygen and pH levels using optical biosensors. With the instrument’s 96 wells, researchers can test the effects of up to four drugs on cellular metabolism, elucidating the bioenergetics of the cell. Currently selling for $100,000 to $200,000, the machine was first released to users in January 2009, and now boasts more than 400 clients worldwide.

WILEY: I want one!

PISTON: This is a great example of how the reduced volumes made possible with microfluidic principles can increase both signal-to-noise and temporal resolution.

 

New sequence capture tool

20091203-6

Scientists have a plethora of invaluable genomic data-3 billion base pairs’ worth-but no way to use it. The genome has been too large and cluttered for researchers to fully analyze the information. Now HybSelect, launched by the Germany-based company febit in March, uses DNA microarrays to narrow in on regions of the genome that play an important role in a particular disease. The technology has already been used to study cancer, multiple sclerosis, Alzheimer’s, and diabetes.

“It lets us dissect a large genome and isolate the juicy bits that can be used to research diseases,” says Peer Stähler, febit’s chief scientific officer and a former microbiologist at the Max Planck Institute for Brain Research.

Researchers interested in isolating specific DNA sequences have two options: they can either send their samples to febit or buy the HybSelect technology themselves. Samples isolated at febit are sent back to researchers with tips on how to best sequence the genes. In case researchers don’t have access to sequencing equipment, the company also offers next-generation sequencing, the whole process taking just 2 weeks and costing as little as $10,000 (for a pilot study), says Stähler. Labs interested in cutting down shipping time can also purchase a Geniom RT Analyzer, the company’s all-in-one microarray processing and analysis instrument, and Geniom Biochip, which contains the HybSelect application, for $150,000. The machine is relatively small (55.7 x 90.7 x 66.5 cm; 110 kg) and can process up to 16 samples a day.

WANG: The idea of sequence capture is not new, but the technological development is new and will improve the capacity and efficiency of deep sequencing.

LEVY: The ever-increasing output of DNA sequencing technologies, the successes of genome-wide association studies, and the appreciation of how rare variants contribute to disease and phenotype all illustrate the need for efficient and cost-effective methods to capture genomic regions of interest for further characterization.

 

All-in-one microscopes

20091203-7

This year saw the introduction of two new all-in-one microscope systems from Olympus: the FluoView FV10i, the world’s first self-contained confocal microscope, which can be used for creating 3D views of a specimen, and the FSX100, a self-contained fluorescence and brightfield microscope, the first of its kind commercially available in the United States. Both systems combine the illumination systems, microscopes, movable stages, and cameras all into a simple little box.

“They don’t look like anything that is typical for scientists,” says Mark Clymer, a product manager for Olympus. The fact that they are self-contained means they “can be installed just about anywhere.” Furthermore, he adds, these systems hold a particular advantage “for fluorescence imaging, which is typically done in dark rooms, [as] it can be done in the laboratories [with] the lights on.”

Video courtesy of Olympus

The FSX100 costs $55,000, and the FluoView FV10i runs $147,000 for the oil-based model and $167,000 for the water immersion version, optimal for live cell imaging.

In addition, both microscopes are completely “software driven,” meaning they are extremely logical and can be easily navigated, even by first-time users. “Someone could sit down and really without any guidance can generate publication-quality images in minutes,” Clymer says, making these microscopes particularly useful in multiuser facilities.

PISTON: Such a simple yet powerful microscope system will expand the use and development of fluorescent protein technology to labs with little or no imaging experience.

WILEY: Very innovative, but will it find a use?

 

Zinc fingers create knockout rat

20091203-8

Sigma-Aldrich took the bronze in last year’s competition for their CompoZr zinc finger nuclease (ZFN) service, which initiates double-strand DNA breaks at specific sites to knock out even a single base pair. This year the company follows up with the first fruit of that platform-the knockout rat.

“We all knew how well CompoZr worked in cell lines, and the natural extension was to use that in vivo,” says Edward Weinstein, director of the company’s Sigma Advanced Genetic Engineering (SAGE) Labs.

This year, Medical College of Wisconsin researchers used custom zinc-finger nucleases from Sigma to create the first targeted knockout rats, some of which glowed green with the expression of a fluorescent protein, such as GFP. Now rodents beyond mice can be developed into models of specific human diseases.

Dave Smoller, president of Sigma’s research biotech business unit, says that Sigma can make custom zinc finger nucleases for $25,000-$35,000, but that as different proteins are validated and “put on the shelf,” the price could come down for some commonly targeted genes. Weinstein said that SAGE Labs aims to sell rat models of human diseases for “a reasonable price,” but declined to be more specific, and will take orders for custom knockout rats. SAGE has already inked a deal with the Michael J. Fox Foundation to create a panel of five different knockout rats that lack genes implicated in Parkinson’s disease.

WILEY: This advancement shows the real power of the ZFN technology. Gene knockouts have proven to be revolutionary in understanding gene function, but have been mostly restricted to mice and simpler model organisms. ZFN technology provides a new approach for making knockouts in a greater variety of organisms.

LEVY: Beyond transgenics, ZFN have numerous applications in basic research, agriculture, and possibly medical therapeutics.

 

A camera that quantifies

20091203-9

Measuring and comparing the level of fluorescence emanating from proteins, capturing co-localization events at membranes, and depicting viral entry are the bread and butter of cell biologists, who often measure these phenomena using electron-multiplying charge-coupled device (EMCCD) cameras. But these devices spit out figures in units of measurement that are essentially arbitrary, dependent on gain settings that can vary from camera to camera or over time. This means that imaging data is basically irreproducible within and across labs.

The Evolve camera, however, makes imaging data quantifiable and reproducible by measuring images in units of photoelectrons, which result when photons from fluorescent proteins or reflected light hit the camera’s sensors. This overlays detailed images with quantitative, standardized data on how many photoelectrons were captured per pixel.

“What we want is for scientists to realize the value of this and start using that unit of measure,” says Deepak Sharma, senior product manager at Photometrics, which released the camera at the end of February.

Sharma won’t say exactly how many Evolves Photometrics has sold so far, but says that the number sold this year is “not in the thousands yet.” Sharma says that the cost of a new Evolve varies according to geography, but that it is “comparable” to EM cameras with a similar CCD, which can go for upwards of $30,000. “We feel that in 4 or 5 years this is going to have changed the direction of imaging science-standardized it.”

WANG: Imagine a world where researchers could reproduce their imaging experiments and more directly compare their data. Just think of the scientific advances we could make if studies were more quantitative and verifiable. And consider the new insights we could derive from being able to integrate data from different experiments.

WILEY: I think this is a great development for quantitative imaging. If supported by software, it could force all cameras to follow.

 

Manipulate cells using light

20091203-10

There’s an ever-growing armament of tools for tagging proteins to watch cellular events unfold, but until recently, researchers lacked ways to experimentally manipulate those events with the same molecular-level precision. A handful of genetically encoded light-sensitive systems have now been reported that do just that, but most require a heavy dose of protein engineeringWendell Lim and his colleagues at the University of California, San Francisco, may have found a solution. Normally, the light-sensitive plant protein phytochrome and its binding partner, phytochrome interaction factor (PIF), link up and translocate to the nucleus in response to red light; infrared light breaks the bond. The researchers modified the genes so that the pair, when activated, instead moved to the cell membrane. They then linked PIF to a cytoskeletal protein. Spatially targeted pulses of red light flipped on PIF, which in turn activated the cytoskeletal protein, reshaping the cell (Nature, 461:997-1001, 2009).

Phytochrome “converts light into a protein-protein interaction,” says Lim. Researchers can link PIF to any number of proteins, potentially making the system applicable to a broader array of cell processes than other light-controlled systems, he adds.

The group submitted the mutant phytochrome and PIF plasmids to Addgene, a nonprofit plasmid repository that facilitates distribution of plasmids among the scientific community. Researchers can request the plasmids for about $65 each.

WILEY: Because the system is genetically encoded, modular (works with any pair of proteins), reversible, and uses nontoxic wavelengths of light, it is likely to have an extremely high impact.

LEVY: This data may offer an unprecedented ability to control protein interaction and localization in the cell.

 

Quick pathogen ID

20091203-11

When facing an outbreak of an unknown, deadly pathogen, any delay costs lives. So in the 1990s, during a government-run meeting on biodefense, David Ecker was disappointed by the best ideas being offered for pathogen detection. “They were talking about the Gram stain,” Ecker recalls.

At the time Ecker, at Ibis Biosciences, had been using mass spectrometry to test drug candidates for their ability to bind to RNA, by comparing the atomic weight of a bound RNA to an unbound (lighter) molecule. He figured, why not use the tool to identify genomes based on their different weights? “If we could measure a small molecule sticking to a nucleic acid, I could just measure a nucleic acid.”

The trick was to design PCR primers for conserved areas in a viral or bacterial genome, making them universal for an entire class of pathogens. The part of the genome sandwiched by the primers and amplified by PCR would be variable enough to distinguish a particular strain and subtype within each class of pathogen.

While it hasn’t been approved for clinical trials or diagnostics yet, the machine is being used for testing basic mutation rates in viruses, forensics, and other applications, including being used by the U.S. Navy and Centers for Disease Control and Prevention to identify the new H1N1 virus.

After their acquisition by Abbott Laboratories late last year, Ibis and Abbott engineers designed a sleeker version of the machine called the PLEX-ID, which the Wall Street Journal dubbed the Innovation of the Year. The tool costs more than $100,000, and $30-$40 per sample.

PISTON: This automated molecular “canary” combines genetics, robotics, spectroscopy, and informatics to greatly accelerate the identification of unknown diseases, and early detection is always important.

WANG: High-throughput detection of infectious agents is a timely development in light of the continuous threats from pandemic agents.

 

Pluripotency from proteins

20091203-12

Protein-induced pluripotent stem cell colonies express endogenous nanog (immunostained in red). Image Courtesy of Hongyan Zhou

This year’s most exciting innovation, announced in April, circumvents the complications that come with the most common technique for reprogramming cells to an embryonic-like state. For the first time, Sheng Ding of Scripps Research Institute in La Jolla, Calif., and his colleagues induced pluripotency in mouse embryonic fibroblast cells using only proteins, avoiding genetic modification altogether.

“The iPS cell technology was really a breakthrough discovery, but genetic modification [poses] tremendous hurdles for practical applications,” including the potential to cause diseases such as cancer, says Ding.

The team struggled with the idea for nearly 2 years before finding the right conditions and the perfect combination of ingredients, which included the protein form of Shinya Yamanaka’s four transcription factors, as well as a histone deacetylase inhibitor known to enhance reprogramming efficiency (Cell Stem Cell, 4(5):381-84, 2009).

San Diego-based Fate Therapeutics, of which Ding is a founder, holds the exclusive license for the protein-induced stem cell technology and the specialized cells derived from it. The technology-which could consist of the solution of proteins with validated protocols or the pluripotent cells themselves-is not commercially available yet, but is being developed “in association with partners,” says Fate CFO Scott Wolchko.

Wolchko declined to comment on the cost other than to say that it will depend on “the ultimate application of the technology,” with the most basic applications such as toxicology testing and the development of reagents at the low end of the price scale, and more advanced drug development and cell therapy applications costing a bit more.

WANG: This study not only overcomes the danger of using transgenes to generate iPS cells, but its result also suggests, to my amazement, that pluripotency, once induced, can be self-propagated without the continuous supply of the exogenous recombinant proteins.

PISTON: Since there is still disagreement about the genetic profile of iPSCs, an alternative derivation of them that preserves their functionality will create new useful cell lines and also lead to better understanding of these cells.

Judge Profiles

JEAN Y.J. WANG, based at the University of California, San Diego, is a distinguished professor in medicine, the chair of the biomedical sciences graduate program, and the associate director of basic research at the Moores UCSD Cancer Center. In studying the functions of cancer genes, her laboratory employs biochemistry, cell biology, molecular biology, mouse genetic models and high- throughput technologies, to elucidate the functional interactions of oncogenes and tumor suppressors in the regulation of differentiation and cell death.

DAVID PISTON is a professor of molecular physiology & biophysics at Vanderbilt University. He is the director of the Vanderbilt Biophotonics Institute, as well as the co-director for biomedical application of Vanderbilt’s Advanced Computer Center for Research and Education (ACCRE). His lab uses quantitative fluorescence microscopy to study living cells and tissues, and he established an in vivo molecular imaging center at Vanderbilt.

H. STEVEN WILEY is the lead biologist at the Environmental Molecular Sciences Laboratory (EMSL) at Pacific Northwest National Laboratory, where he uses cell imaging, computational biology and high-throughput proteomics to understand cell communication. His work combines the techniques of molecular and cellular biology with both biochemical and optical assays, and uses the results to construct computer models of the cellular processes. He sits on the editorial board of The Scientist, where he is also a columnist.

SHAWN LEVY is faculty investigator at the HudsonAlpha Institute for Biotechnology. Prior to joining HudsonAlpha, Levy was an assistant professor of biomedical informatics and molecular physiology and biophysics at Vanderbilt University Medical Center. His research interests include technology and methods development in high-density gene expression profiling, genotyping, structural and functional genomics, and the development of bioinformatic tools for the integration of clinical and molecular data from diverse technology platforms.

20091203-1

The-Scientist.com, December 2, 2009, by Jef Akst  —  Oklahoma State University (OSU) administrators have aborted a federally-funded study of anthrax vaccines because the project involved sacrificing the baboons involved in the research — even though the project had already received approval by a unanimous vote from the university committee overseeing animal research.

“It was a considerable surprise to pretty much everybody involved,” said Michael Davis, an OSU veterinary doctor and a member of the Institutional Animal Care and Use Committee (IACUC) that approved the project. “It’s not as though this was the first time anybody suggested that we ought to euthanize an animal during a research protocol.”

The project, headed by Boston University’s Shinichiro Kurosawa, proposed to use baboons as a primate model to test the efficacy of the current vaccine (the one given to members of the military) for anthrax. The plan was to expose the animals to the spores of the attenuated Sterne strain of anthrax and eventually advance to the Ames strain — the fully encapsulated and virulent form of the bacterium that was used in the anthrax attacks of 2001 — and observe the pathobiology of infection. It was part of a collaborative multi-institutional NIH grant originally awarded for $12 million in 2004, and renewed in September of this year for another $14.3 million.

Kurosawa’s proposed subproject, which had a direct cost budget of $200,000 per year, required special laboratory conditions: future experiments involving the Ames strain, for example, would have to be done in a biosafety level (BSL) 3 facility. The new laboratory at OSU fit the part — a large animal facility with BSL 3 clearance, close proximity to the Oklahoma Medical Research Foundation (OMRF) where Kurosawa used to work, and a baboon colony. Kurosawa filed for permission with the institution’s IACUC, which approved the project on September 15. (The project was still pending review and approval by the Institutional Bio-Safety Committee when the administration made its decision so the IACUC never issued approval letters.)

“The impression that I had, as one of the members of the IACUC, is that we were the last step,” Davis said. “Everything else was essentially in place.” But before final permission was granted and he was able to begin his study, Kurosawa received an email from OSU vice president of research Stephen McKeever saying that OSU was unwilling to host it, reported The Oklahoman — who, it seems, first reported the story (Hat tip – DrugMonkey).

OSU administrators declined to comment, but did release a statement saying that the proposed research “was not in the best interest of the university” and that it “would have distracted from [ongoing] efforts.”

But the project proposed by Kurosawa is exactly the type of research the new OSU facility was built for, OSU veterinary scientist Richard Eberle wrote in an email to The Scientist. The new lab was intended not just for OSU researchers; in fact, the Oklahoma University Health Science Center opted not to build a BSL3 primate facility since this one would be available at OSU, said Eberle, the OSU principal investigator for the proposed research. “So one of the things that I find most chilling about this decree,” he said, “is that it will not only shut OSU researchers out of this type of research, but will also exclude researchers from other institutions in Oklahoma and elsewhere in the US.”

“We were surprised,” said immunologist K. Mark Coggeshall of OMRF, the PI of the collaborative NIH grant. “We’re disappointed, but we understand — these are philosophical distinctions.”

Coggeshall called the university’s decision a “delay” to their research, and said that they plan to continue their work with baboons. “We’ll just find another site,” he said. But that may be easier said than done, warned Davis. “The type of facility you need to do this is not exactly on every street corner,” he said. Indeed, the new OSU facility is the only place in the state that has an animal BSL 3 facility and only “one of a few such facilities in the US” that can host primate research on biological toxins, agreed Eberle.

Some OSU researchers expressed concern about the precedent set by the cessation of this project for future studies involving animal subjects. “Personally I’m still not absolutely certain where the policy lines are,” Davis said. Based on this decision, it seems that “the status quo is that the university has banned terminal primate research,” he added. “At the very least, if there’s going to be a policy going forward, that policy needs to be clear.”

The issue will be presented to the OSU Faculty Council next week, Eberle said. “I have no idea where things will go from there.”

Kurosawa declined to comment.

20091203-2

Anthrax bacteria (blue rods) in cerebrospinal fluid from the first case of inhalational anthrax due to bioterrorism in the United States, 2001. (Courtesy of JFK Medical Center, Atlantis, Fla.)

December 2, 2009, by Gabe Mirkin MD  —   Sit-ups can strengthen your belly muscles, but doing them incorrectly can hurt your back. Sit-ups should be done while you lie on your back with your knees bent enough for the soles of your feet to touch the floor. Place both hands on your chest and slowly raise your head off the ground. Raise your shoulders about one foot and then lower them to the ground. Do this slowly ten times, rest a few seconds and then do two more sets of ten. After a week or two this exercise will feel easy, so add a light weight held behind your neck or on your chest. As you become stronger, you can use heavier weights.

There’s no need to do more than 30 sit-ups in one workout. To strengthen your belly muscles, you increase the resistance, not the number of repetitions. Keep your knees bent to protect your back. If you do a sit-up with your legs straight, you place a great force on the iliopsoas muscles that increase the arch in your back, which can damage the ligaments and joints. If your belly muscles are weak, you are likely to arch your back excessively when you sit up and increase the chances of injury. If you are doing sit-ups to flatten your stomach, you need to raise your head only about one foot because going higher than that uses the quadriceps muscles in the front of your upper legs, not your belly muscles.