New tests for 'legal marijuana,' 'bath salts' and other emerging designer drugs

 

Scientists today reported development of much needed new tests to help cope with a wave of deaths, emergency room visits and other problems from a new genre of designer drugs sold legally in stores and online that mimic the effects of cocaine, ecstasy and marijuana. They spoke at the 242nd National Meeting & Exposition of the American Chemical Society (ACS).

The reports, among more than 7,500 on the ACS agenda, focus on drugs sold as "bath salts," "plant food," "incense" and other products with colorful names, such as "Ivory Wave," "Red Dove" and "legal marijuana." They provide users with a high, but many have not yet been made illegal and are undetectable with current drug tests. In one presentation on these "legal highs," a United Kingdom researcher reported a new method to trace the source of the substances in "bath salts." In the other, a U.S. researcher discussed the challenges facing law enforcement and policy makers in regulating synthetic versions of marijuana.

Oliver Sutcliffe, Ph.D., and his collaborators reported the successful use of a method called isotope ratio mass spectrometry (IRMS) to determine who is making bath salts — drugs that can cause euphoria, paranoia, anxiety and hallucinations when snorted, smoked or injected — and which chemical companies supplied the raw materials. He and his co-workers are based at the University of Strathclyde and the James Hutton Institute in the U.K.

"With the new method, we could work backwards and trace the substances back to the starting materials," said Sutcliffe. IRMS measures the relative amounts of an element's different forms, or isotopic ratio. "This method was successful because the isotopic ratio of the starting material is transferred like a fingerprint through the synthesis," he explained.

"Bath salts" first garnered major media attention in the U.K. in early 2010, and then became a problem in the U.S. These products are not in the supermarket soap aisle — they are sold on the Internet, on the street and in stores that sell drug paraphernalia. They are sold in small individual bags for as low as $20 each for the real purpose of providing a cheap, legal high.

The powders often contain mephedrone, which is a synthetic compound, structurally related to methcathinone, which is found in Khat — a plant that is illegal in many countries, including the U.K. and the U.S. Usually, that would mean that these compounds (and derivatives thereof) would be illegal in those countries too, but because the bath salts are labeled "not for human consumption," they get around this restriction and other legislation governing the supply of medicines for human use. However, Florida and Louisiana — two hotspots of bath salts abuse — specifically banned the substances. U.K. officials banned the import of bath salts, which may lead some in the drug trade to set up clandestine labs on U.K. soil, said Sutcliffe. The new method provides law enforcement with a tool to track down these bath salts manufacturers.

In previous work, Sutcliffe developed the first pure reference standard for mephedrone, as well as the first reliable liquid chromatography test for the substance, which could be easily run in a typical law enforcement lab. The team is also developing a color-change test kit for mephedrone, which he estimates may be available by the end of the year.

In another presentation, Robert Lantz, Ph.D., from the Rocky Mountain Instrumental Laboratories, described another high that is legal in most of the U.S. — synthetic cannabinoids marketed as incense, a spice product or "legal marijuana" that give a high similar to marijuana without showing up in conventional drug tests.

"We can detect synthetic cannabinoids with modern analytical chemistry techniques, such as liquid or gas chromatography coupled to mass spectrometry, but these assays are too expensive for the 5,000-10,000 urine samples that most drug testing labs receive each day," said Lantz. Most labs screen for drugs with less expensive antibody assays, but because the structures of these substances are so dissimilar, different antibodies would likely be required for many of them, driving up the cost of a more comprehensive test.

Synthetic cannabinoid abuse rose sharply in 2010, according to U.S. poison control centers, up to 2,863 compared to only 14 in 2009. About 200 synthetic cannabinoids exist, but the U.S. Drug Enforcement Agency (DEA) banned only five of those. A handful of states, such as Washington, Georgia and Colorado, banned five of them, but they are not always the same five that the DEA banned. "The states banned several specific compounds without a particular basis for their choices," Lantz pointed out.

Colorado recently passed a law banning any substance that binds to a cannabinoid receptor in the human body. "The bill was well-intentioned, but technically, the new law not only covers synthetic cannabinoids, but also endocannabinoids, which are naturally occurring substances that the human body produces to regulate many normal processes," said Lantz.

Provided by American Chemical Society (news : web)

Cars could run on recycled newspaper, scientists say

Here's one way that old-fashioned newsprint beats the Internet. Tulane University scientists have discovered a novel bacterial strain, dubbed "TU-103," that can use paper to produce butanol, a biofuel that can serve as a substitute for gasoline. They are currently experimenting with old editions of the Times Picayune, New Orleans' venerable daily newspaper, with great success.

TU-103 is the first bacterial strain from nature that produces ?butanol directly from cellulose, an organic compound.

"Cellulose is found in all green plants, and is the most abundant organic material on earth, and converting it into butanol is the dream of many," said Harshad Velankar, a postdoctoral fellow in David Mullin's lab in Tulane's Department of Cell and Molecular Biology. "In the United States alone, at least 323 million tons of cellulosic materials that could be used to produce butanol are thrown out each year."?

Mullin's lab first identified TU-103 in animal droppings, cultivated it and developed a method for using it to produce butanol. A patent is pending on the process.

"Most important about this discovery is TU-103's ability to produce butanol directly from cellulose," explained Mullin.

He added that TU-103 is the only known butanol-producing clostridial strain that can grow and produce butanol in the presence of oxygen, which kills other butanol-producing bacteria. Having to produce butanol in an oxygen-free space increases the costs of production.

As a biofuel, butanol is superior to ethanol (commonly produced from corn sugar) because it can readily fuel existing motor vehicles without any modifications to the engine, can be transported through existing fuel pipelines, is less corrosive, and contains more energy than ethanol, which would improve mileage. ?

"This discovery could reduce the cost to produce bio-butanol," said Mullin. "In addition to possible savings on the price per gallon, as a fuel, bio-butanol produced from cellulose would dramatically reduce carbon dioxide and smog emissions in comparison to gasoline, and have a positive impact on landfill waste."

Provided by Tulane University

Cracking cellulose: a step into the biofuels future

Scientists from the University of York have played a pivotal role in a discovery which could finally unlock the full potential of waste plant matter to replace oil as a fuel source.

Professor Paul Walton and Professor Gideon Davies, of the University's Department of Chemistry, were part of an international team that has found a method to overcome the chemical intractability of cellulose, thus allowing it to be converted efficiently into bioethanol.

Working with scientists in Novozymes laboratories at Davis, California, and Bagsvaerd, Denmark, as well as researchers at the University of Copenhagen and the University of Cambridge, they identified the molecular mechanism behind an enzyme found in fungi which can degrade the cellulose chains of plant cell walls to release shorter sugars for biofuels.

This represents a major breakthrough as cellulose is the world's most abundant biopolymer. Global generation of cellulose is equivalent in energy to 670 billion barrels of oil – some 20 times the current annual global oil consumption. The discovery opens the way for the industrial production of fuels and chemicals from plentiful and renewable cellulose in waste plant matter.

The research, which is published in the Proceedings of the National Academy of Sciences (PNAS), removes the major constraint on the production of bioethanol from cellulose the stability of which had previously thwarted previous efforts to make effective use of it for biofuels.

The researchers found a way of initiating effective oxidative degeneration of cellulose using the copper-dependent TaGH61 enzyme to overcome the chemical inertness of the material.

Professor Davies, much of whose work on plant cell-wall degradation is funded by the Biotechnology and Biological Sciences Research Council, said: "Cracking cellulose represents one of the principal industrial and biotechnological challenges of the 21st century. Industrial production of fuels and chemicals from this plentiful and renewable resource holds the potential to displace petroleum-based sources, thus reducing the associated economic and environmental costs of oil and gas production. Events at Fukushima and the continuing instability in major oil producing countries only highlight the need for a balanced energy portfolio."

Professor Walton added: "This discovery opens up a major avenue in the continuing search for environmentally friendly and secure energy. The potential of bioethanol to make a major contribution to sustainable energy really now is a reality."

Claus Crone Fuglsang, Managing Director at Novozymes' research labs in Davis, California said: "Scientists have worked to figure out how to break down plant matter for the past 50-60 years. The impressive effect of GH61 was established a few years back and today it is a key feature of our Cellic CTec products.

"Fully understanding the mechanism behind GH61 is important in the context of commercial production of biofuel from plant waste and a true scientific paradigm shift. This discovery will continue to drive advances in production of other biobased chemicals and materials in the future."

Leila Lo Leggio, Group Leader of the Biophysical Chemistry Group at the Department of Chemistry, University of Copenhagen, said: "As a team of academic scientists, it is particularly rewarding when our basic research in the three-dimensional structure and chemistry of proteins also contributes to possible solutions for one of the major challenges our society is facing."

Professor Paul Dupree of the University of Cambridge Bioenergy Initiative and Director of the BBSRC Sustainable Bioenergy Cell Wall Sugars programme, said "Understanding the GH61 enzyme activity is one of the most significant recent advances in the area of biomass deconstruction and release of cell wall sugars."

Provided by University of York

Controlling cells' environments: A step toward building much-needed tissues and organs

 

With stem cells so fickle and indecisive that they make Shakespeare’s Hamlet pale by comparison, scientists today described an advance in encouraging stem cells to make decisions about their fate. The technology for doing so, reported here at the 242nd National Meeting & Exposition of the American Chemical Society (ACS), is an advance toward using stem cells in “regenerative medicine” -- to grow from scratch organs for transplants and tissues for treating diseases.

“Stem cells have great potential in regenerative medicine, in developing new drugs and in advancing biomedical research,” said Laura L. Kiessling, Ph.D., who presented the report. “To exploit that potential, we need two things: first, reproducible methods to grow human stem cells in the laboratory, and second, the ability to make stem cells grow into heart cells, brain cells or whatever kind of cell. Our technology takes a different approach to both of these problems, and the results are very encouraging.”

Biologically, so-called pluripotent human embryonic stem cells have not made up their minds about what to become. That’s essential because these cells, which are derived from embryos, have the agility to develop into the hundreds of different kinds of cells in a fully-formed human body. But controlling their differentiation has also stood as a major barrier to making the stem cell dream come true and using these all-purpose cells in medicine.

Past approaches to growing and scripting the fate of stem cells have involved adding growth-regulating and other substances to cultures of stem cells growing in the laboratory. These conditions left scientists guessing about exactly what wound up in the stem cells. Kiessling and colleagues are pioneering a new approach that involves using chemically controlled surfaces.

Kiessling previously developed chemically modified plastic and glass surfaces that take much of the guess work out of growing stem cells in laboratory cultures. In the past, scientists grew stem cells on surfaces that contained mouse cells. That left scientists with nagging questions about possible contamination of stem cells with disease-causing animal viruses — a stumbling block for using stem cells in potential medical applications. And that growth system was what scientists term “undefined.” There were variations from batch to batch of mouse cells, and scientists never really knew what the stem cells were coming into contact with and how it might be changing them. The synthetic, chemically-defined, surfaces ended that uncertainty. The approach was inexpensive, simple and a much-needed advance in producing stem cells, Kiessling explained.

With the ability to grow stem cells on the synthetic surfaces under chemically defined, or known, conditions, Kiessling’s group took an additional step in their latest research. It found that chemically defined surfaces can exert control over signaling pathways. “Signaling” is how molecules talk to one another and get things done inside a cell. It’s how an immune cell knows to fight an infection or how a pancreatic cell determines that more insulin is needed in the bloodstream, for example. By controlling how molecules inside a stem cell communicate, researchers could someday in the future nudge them to become one type of cell or tissue over another.

To see whether a new chemically defined surface could change signaling in a pilot experiment, Kiessling tested cancer cells. The research involved use of a signaling substance, transforming growth factor-beta (TGF-beta), which controls a range of activities, from cell growth to self-destruction.

“The new surfaces give scientists much more control over cells, opening up a wide range of possible future applications,” Kiessling explained. Building directly on the results of the pilot study, the surfaces could have applications in wound healing. TGF- beta can help wounds heal, but if it touches healthy skin, inflammation or even a cancerous tumor could develop. “We haven’t done this, but you could imagine a bandage that has a localized concentration of the special peptide surface that would recruit TGF-beta just to the wound site,” said Kiessling.

The surfaces also could make it easier to manufacture organs and tissues in the laboratory someday. “We think that this strategy, with different sets of peptides (building blocks of proteins) bound to the surface, could direct certain human embryonic stem cells on the surface to become one type of cell and other stem cells to become a second cell type, right next to each other. For the tissue engineering involved in growing replacement organs, you need to organize specialized cells in particular ways like this.”

Provided by American Chemical Society (news : web)

Research offers new way to target shape-shifting proteins

A molecule which can stop the formation of long protein strands, known as amyloid fibrils, that cause joint pain in kidney dialysis patients has been identified by researchers at the University of Leeds.

The discovery could lead to new methods to identify drugs to prevent, treat or halt the progression of other conditions in which amyloid fibrils play a part, including Alzheimer's, Parkinson's and Type II diabetes.

The research, funded by the Biotechnology and Biological Sciences Research Council and the Wellcome Trust, is published today (August 28) in Nature Chemical Biology.

The team – from Leeds' Astbury Centre for Structural Molecular Biology and Faculty of Biological Sciences – found that an antibiotic known as Rifamycin SV was able to prevent the protein ß2microglobulin (ß2m) from forming into fibrils. ß2m is known to accumulate in renal dialysis patients and forms fibrils within the joints, causing extreme pain and arthritis.

By using a specialised analytical technique called ion mobility spectrometry-mass spectrometry (IMS-MS), the researchers were able to see at what stage of the process Rifamycin SV prevented amyloid fibril formation. They believe the technique could enable potential drugs to be identified for the many other proteins which form amyloid fibrils, linked to a wide range of human disorders.

"Traditional drug design for diseases like Alzheimer's is incredibly difficult because the proteins you're trying to target are changing shape and structure all the time," explains University of Leeds Professor of Structural Molecular Biology, Sheena Radford. "It's like trying to consistently pick out one bead of a particular shape from box of potentially millions of similar beads. This new technique allows us to see the shape of the protein as it changes, so we can more easily identify exactly which part we need to target."

In their normal, folded state, proteins are unable to link together to form long fibrillar assemblies, but if they unfold, they expose areas where they can bind to each other. Initially they form small groups of two, three or four proteins, and then these link into long strands, which twist together to form fibrils.

Most analytical techniques can only show the mass of the protein or its make-up in terms of amino acids, neither of which changes as the protein unfolds. Others are unable to look at individual molecules within complex mixtures. However, IMS-MS can measure the mass and shape of a protein, allowing researchers to watch the unfolding process and the aggregation into small groups and then assembly into the fibril and to find which of these species is able to bind a ligand and stop the assembly process.

In the research published today, researchers found that Rifamycin SV stopped the formation of protein fibrils by binding to an unfolded protein molecule with a particular shape, enabling for the first time, an unfolded protein of a particular shape to be identified as a target for the design of new inhibitors of fibril assembly.

"We're fortunate to be one of the few universities in the UK able to use IMS-MS to study amyloid fibril formation," says Professor of Biomolecular Mass Spectrometry, Alison Ashcroft, who specialises in this type of analysis. "Although fibrils take years to develop in the body, we are able to 'grow' them in hours in the lab. By using IMS-MS to help us map exactly how they are formed, we can better understand the mechanism by which it happens and – we hope – find ways to stop it."

More information: Ligand binding to distinct states diverts aggregation of an amyloid-forming protein is published on August 28 via Advance Online Publication (AOP) on the Nature Chemical Biology website. DOI: 10.1038/NChemBio.635

Provided by University of Leeds (news : web)