The Story of Digitalis Digitalis is an example of a cardio-active or cardiotonic drug, in other words a steroid which has the ability to exert a specific and powerful action on the cardiac muscle in animals, and has been used in the treatment of heart conditions ever since its discovery in 1775. The discovery of digitalis is accredited to the Scottish doctor William Withering, and makes for quite an interesting historical story. In 1775, one of his patients came to him with a very bad heart condition and since Withering had no effective treatment for him, thought he was going to die. The patient, being an independent type, went instead to a local gypsy, took a secret herbal remedy - and promptly got much better! When Withering heard about this, he became quite excited and searched for the gypsy throughout the by-ways of Shropshire. Eventually he found her, and demanded to know what was in the secret remedy. After much bargaining, the gypsy finally told her secret. The herbal remedy was made from a whole concoction of things, but the active ingredient was the purple foxglove, digitalis purpurea. The potency of digitalis extract had been known since the dark ages, when it had been used as a poison for the mediaeval 'trial by ordeal', and also used as an external application to promote the healing of wounds. There are also reports of digitalis extract finding some use in the treatment of dropsy. So, Withering tried out various formulations of digitalis plant extracts on 163 patients, and found that if he used the dried, powdered leaf, he got amazingly successful results. He introduced its use officially in 1785. Even today, drugs based on digitalis extract, such as Digitoxin and Digoxin, are some of the best known treatments to control the heart rate. It works by increasing the intensity of the heart muscle contractions but diminishing the rate, and doses as low as 0.3mg daily are all that is needed. Since digitalis purpurea contains a mixture of several cardiac glucosides and also several saponins in amounts and proportions which vary with locality and with season, digitalis preparations vary considerably in potency and quality. Because of this, and the fact that the therapeutic dose is so small, it is very easy to exceed the safe dosage. Indeed, Withering recommended that the drug be diluted and administered repeatedly in small doses until a therapeutic effect became evident. This procedure was very effective in experienced hands, but also very time-consuming. Nowadays, therefore, preparations from digitalis leaves are made using modern recrystallisation methods and are carefully standardised by bio-assay. Source: http://www.bristol.ac.uk/Depts/Chemistry/MOTM/digitalis/digtalis.htm and Local Heroes, BBC2 TV Programme, 1996

Biopiracy fears hampering research in Brazilian Amazon By Michael Astor, Associated Press

October 30, 2005

RIO DE JANEIRO, Brazil — Somewhere in the Amazon there may be flora and fauna that hold the key to curing diseases ranging from cancer to multiple sclerosis. That, at any rate, is the dream. But the reality is that the search for the next miracle drugs is being hampered by a deep Brazilian suspicion of "biopiracy." Rubber trees in southeast Asia. Brazil blames the British for illegally smuggling rubber seeds out of the Amazon rainforest in the late 19th century to establish plantations in Ceylon (Sri Lanka) and Malaysia. These plantations eventually cut Brazil's share of the rubber market from a virtual monopoly to 20% in 1918. By 1940, Brazil's share had fallen to 1.3% of the world rubber market.

Some politicians, retired generals and Web sites seem convinced that the world's biggest rain forest is crawling with biopirates scooping up seeds, leaves and animal blood samples whose genetic code might deliver the next miracle drug. The government has imposed strict regulations which apply to both Brazilians and foreigners, but foreigners are more likely to get arrested. Over the past decade more than 30 have been detained, and their research samples confiscated or destroyed. The Amazon rain forest is thought to contain at least 30 percent of all plant and animal species on the planet, most of them uncatalogued. At the same time, loggers and farmers are shrinking its area at a rate equivalent to six football fields a minute. But scientists say the rules are so stringent and overzealously enforced that it has become impossible to ship samples abroad for analysis, reducing research to a crawl and driving many scientists to move their research to Ecuador, Bolivia and Peru. Last year, police tracked two German researchers across eight Brazilian states and seized the spiders they were allegedly planning to ship to the United States. In 2002, Marc Van Roosmalen, a Dutch scientist who has discovered some 20 new monkey species, was accused of biopiracy after authorities removed 27 rare monkeys from his home in the Amazon city of Manaus. Roosmalen says he was only studying and caring for the animals, not exploiting them for profit, and had applied for permits in 1996 and never heard back. Brazilian scientists are feeling the squeeze too. "The situation is so frustrating, I've all but given up," says Paulo Buckup, a professor of ichthyology at the Federal University of Rio de Janeiro who collects river fish for research. "Brazil has lost the capacity to control its own resources because it doesn't know what it has." Biopiracy haunts Brazilian history, beginning with Henry Wickham, an Englishman who smuggled rubber seeds out of the country in the 19th century and broke Brazil's global rubber monopoly.

Then came the 1992 Earth Summit in Rio de Janeiro, which produced a convention entitling nations to a share of the profits from substances yielded by their flora and fauna. "All the signers bought into a concept no one knows how to implement. Anyone can claim you're not sharing the benefits, and the government is afraid of being held responsible," said Dr. Roberto Cavalcanti, a zoology professor at the University of Brasilia. Cavalcanti agrees regulation is necessary, but feels the best way to fight biopiracy is more investment and more Brazilians doing their own collecting. He also says the biopiracy concept "has been hijacked" by opponents of measures to protect the rain forest against commercial overexploitation. A congressional committee is investigating biopiracy, and several prominent foreign scientists have been forced to prove they are not biopirates, including Thomas Lovejoy, the U.S. scientist credited with putting the plight of the rain forests on the world's radar screen in the early 1980s. He acknowledges he shares the blame for the biopiracy panic because of his own role in publicizing biodiversity. "From my point of view, the real biopiracy is the destruction of the biodiversity of the Amazon," said Lovejoy, president of the Heinz Center for Science Economics and the Environment. Lovejoy was eventually cleared of vague charges that he was a CIA agent when he did research for the Smithsonian Institute in the Amazon years ago, and Congressman Jose Sarney Filho, a former environment minister on the biopiracy committee, acknowledges the investigation so far has little to show for its work. "Up to now, we haven't found a single concrete case of biopiracy," Sarney told The Associated Press. "There are cases of spiders being contrabanded to American laboratories and things like that, but no material proof that our flora or fauna has been converted into medicine without following the legislation." But that doesn't silence the cries of alarm. "The internationalization of the Amazon goes far beyond the economic area and the occupations of lands," Amazonas state Gov. Eduardo Braga warns. "They will take from us our flora and our fauna." Manaus Mayor Serafim Correa says Brazilians must "take care that we don't allow our Amazon to be invaded." On the Web site "Amazon Love it or Leave It," Gen. Luiz Gonzaga Schroeder Lessa, former chief of the Amazon Military Command, claims collectors disguised as religious or environmental groups are taking samples to be turned into medicines for which Brazilians will later have to pay them royalties. "It's biopiracy and it goes on almost unchecked in the Amazon," he writes. Sarney, the congressman, says most Brazilians confuse biopiracy with things like a recent case where a Japanese company trademarked "Cupuacu," a fruit unique to the Amazon. The trademark was revoked following protests from Brazil. Rogerio Magalhaes, an environment ministry official, acknowledges the bureaucracy is frustrating, but denies it stops researchers from doing research. "They're doing it but they're doing it illegally," he says. The legal limbo provides little comfort to scientists like Carlos Joly, director of the Botanical Institute at the University of Campinas in Sao Paulo state. "Right now it seems like we — the ones who are doing research — are the pirates," said Joly. "The best way to protect Brazil's biodiversity is to know its characteristics and potential. That's what the country should be investing in."

Venomous Snails Offer a Weapon to Kill Pain By Robin McKie The Observer 4/27/08

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Sea snails that hunt fish by spitting tiny venom-tipped harpoons at them are being used to develop life-saving medicines. One group of scientists has already succeeded in pinpointing a new long-lasting anaesthetic and others have created a powerful, non-addictive painkiller from the snail poison. “There are several hundred different species of cone snail and all use poisons to kill their prey,” said Professor Alan Harvey of Strathclyde University's institute of drug research. 'In turn, each species uses poison made up of several hundred different compounds which offers us, in total, an extremely wide range of chemicals to test and develop.' His team is the only British group involved in the £8m EU-funded Cone Snail Genome Project for Health, Conco, that has been set up specifically to study cone snails, isolate their toxins and exploit those that make the most promising medicines. Scientists believe that cone snail toxins offer particular promise in the development of: · drugs for treating stroke victims; · powerful painkillers; · medicines for controlling the effects of diabetes; · new anti-microbial agents. Cone snails live in seas worldwide. However, the largest ones are found in warm tropical waters, including those of the Indian and Pacific Oceans. All have twisted shells shaped like icecream cones, although individuals rarely grow bigger than a few inches. Scientists now know cone snails are hunters: some kill molluscs, others attack fish. A classic example of their predation is provided by the snail type Conus consors, which is found in the Pacific Ocean. “It's hunting is spectacular,” said Harvey. “When a snail detects a fish, it loads a little tip at the end of a proboscis - it's like a microscopic harpoon - with venom and fires it by a powerful muscular contraction. The venom contains a cocktail of toxins, called conotoxins, which instantly paralyses and kills the fish. The snail then crawls over and engulfs its prey.” Researchers working for the Conco project, which is being co-ordinated by Dr Reto Stöcklin at the Atheris Laboratories in Geneva, have already isolated one toxin from Conus consors which has shown considerable potential as a long-lasting local anaesthetic. Development work on this drug is now being carried out at several laboratories. As part of the project, scientists intend to sequence the entire genome of Conus consors and identify every chemical in the complex cocktail of drugs that makes up its venom. In addition, marine biologists are seeking out new species of cone snail - in the Indian and Pacific Oceans - which would provide further sources of drugs. However, the cone snail drug that has reached the most advanced stage of development is ziconotide, developed by Professor Baldomero Olivera at the University of Utah. It is 1,000 times more potent than morphine but is not addictive. It is aimed at people suffering from severe, chronic pain, experienced by those with severe arthritis or head injuries, for example, and is a synthetic version of the venom used by Conus magus, the Magician's Cone Snail. Olivera and his colleague Michael McIntosh identified one venom component that blocks the calcium channels on the nerves that transmit pain signals. Once the channels are blocked, calcium cannot enter the cells and pain signals are prevented from traveling between nerve cells. “There have been amazing stories told about people taking ziconotide during trials - of people who couldn't walk because of nerve damage to their legs but who were able to dance after a few weeks of taking the stuff,' said Harvey. 'That shows the potential of this approach and indicates the kind of discovery we want to replicate.”

GENOMIC SCREEN NETS HUNDREDS OF HUMAN PROTEINS EXPLOITED BY HIV FINDINGS: Using a technique called RNA interference to screen thousands of genes, researchers identified 273 human proteins required for HIV propagation. The vast majority had not been connected to the virus by previous studies. RELEVANCE: Current drugs attack HIV itself, leaving patients vulnerable to counterattack by the rapidly mutating virus, which often evolves resistance. But the human proteins exploited by HIV represent potential therapeutic targets that could avoid this problem. The challenge will be to develop drugs that inhibit HIV by interacting with these human proteins without hurting our cells. Dr. Stephen Elledge BOSTON, Mass. (January 10, 2007) — In some ways, HIV resembles a minimalist painter, using a few basic components to achieve dramatic effects. The virus contains just nine genes encoding 15 proteins, which wreak havoc on the human immune system. But this bare bones approach could have a fatal flaw. Lacking robust machinery, HIV hijacks human proteins to propagate, and these might represent powerful therapeutic targets. Postdoctoral researcher Abraham Brass (shown here in the Institute of Chemistry and Cell Biology at Longwood) and colleagues identified 273 human proteins required for HIV propagation by screening thousands of genes. Using a technique called RNA interference to screen thousands of genes, Harvard Medical School researchers have now identified 273 human proteins required for HIV propagation. The vast majority had not been connected to the virus by previous studies. The work appears online in Science Express on Jan. 10. Drugs currently used to treat the viral infection interact directly with the virus itself, and it’s quite simple for the rapidly mutating virus to avoid destruction by altering how it interacts with these chemicals. Patients use a cocktail of HIV inhibitors because the virus is less likely to evolve resistance to multiple drugs at the same time. But some HIV strains have still managed to evade particular drugs. These could eventually develop resistance to several drugs, especially among patients who don’t adhere to their regimens. “Antiviral drugs are currently doing a good job of keeping people alive, but these therapeutics all suffer from the same problem, which is that you can get resistance, so we decided to take a different approach centered on the human proteins exploited by the virus,” says Harvard Medical School (HMS) Professor and senior author Stephen Elledge, who holds primary appointments in the Department of Genetics and at Brigham and Women’s Hospital. “The virus would not be able to mutate to overcome drugs that interact with these proteins.” Labs around the world have made impressive contributions to our understanding of the HIV life cycle. Over the last two decades, they’ve identified dozens of human proteins, or host factors, required for HIV propagation. The new study builds on this work, essentially quadrupling the list of host factors to include proteins involved with a surprising array of cellular functions ranging from protein trafficking to a type of programmed cell death called autophagy.

“The expanded list is a hypothesis generation machine,” explains Elledge, who is also a member of the HMS-Partners Health Care Center for Genetics and Genomics and investigator with the Howard Hughes Medical Institute. “Scientists can look at the list, predict why HIV needs a particular protein, and then test their hypothesis.” He hopes that such research will lead to new therapeutics. To create the list, postdoctoral researcher and first author Abraham Brass—working with Derek Dyxkhoorn and Nan Yan from HMS Professor Judy Lieberman’s lab—began with a library of short interfering RNAs (siRNAs) targeting specific human genes. Each siRNA disrupts the gene’s ability to produce a particular protein. With the help of the staff at the Institute of Chemistry and Cell Biology at Longwood (ICCB-L), Brass placed the siRNAs on thousands of human cells, with just one gene being targeted in each well of cells. Thus each well contained cells lacking a particular protein. Next, he unleashed HIV on the cells. If HIV replication was inhibited in a given well, it suggested the missing protein was involved. Of the 273 proteins he identified, just 36 had been previously implicated in the HIV life cycle. He picked three of the other 237 proteins, and subjected them to a host of careful genetic experiments, proving they too truly play a role in HIV propagation. Immune cells—the very cells HIV attacks—contain high concentrations of many of the 273 host factors, offering further proof of the list’s validity. “We’re closing in on a systems level understanding of HIV, which opens new therapeutic avenues,” says Elledge. “We might be able to tweak various parts of the system to disrupt viral propagation without making our own cells sick.” “This is the first whole genome screen for human proteins required by HIV, and we’re confident that it netted real results,” adds Brass. “Given the method, we missed some proteins, but the majority of the ones we found are highly likely to play a role in HIV propagation.” Published on FierceBiotech (http://www.fiercebiotech.com) Source URL: http://www.fiercebiotech.com/press-releases/genomic-screen-nets-hundreds-human-proteinsexploited-hiv CITATION: Brass, et al.(2008) Identification of Host Proteins Required for HIV Infection Through a Functional Genomic Screen. 319: 921-6.

'Non-­stick'  frog  protein  spawns  new  antibiotics   New  Scientist,  25  August  2007     What  do  you  get  if  you  cross  a  frog  with  a  non-­‐stick  frying  pan?  Possibly  a   new  generation  of  antibiotics.     For  years,  researchers  have  tried  to  make  antibiotics  from  antimicrobial   peptides  (AMPs)  -­‐  proteins  found  in  almost  all  animals,  but  particularly  in  the   skin  of  the  African  clawed  frog,  Xenopus  laevis.  AMPs  provide  the  first  line  of   defence  against  invading  bacteria,  but  when  injected  into  the  body,  they  are   quickly  degraded  by  enzymes  called  proteases.  If  the  concentration  of  AMPs  is   increased  to  overcome  this  they  become  toxic,  sticking  to  red  blood  cells  and   killing  them.     Now  Neil  Marsh  and  his  colleagues  at  the  University  of  Michigan,  Ann  Arbor,   have  discovered  that  adding  fluorine  atoms  -­‐  the  key  component  of  the  non-­‐ stick  coating  Teflon  -­‐  to  AMPs  protects  them  from  proteases.     By  swapping  some  of  the  amino  acids  in  the  peptide  for  ones  containing   fluorine,  they  created  an  AMP  that  remained  intact  after  exposure  to   proteases  for  10  hours.  AMPs  without  fluorine  were  completely  degraded   after  30  minutes.     Fluorine  also  made  the  AMPs  more  potent  against  certain  bacteria,  including   MRSA.  The  findings  were  presented  at  the  American  Chemical  Society   meeting  in  Boston  this  week.     The  researchers  are  now  investigating  whether  fluorinated  AMPs  are  less   likely  to  stick  to  and  destroy  red  blood  cells.  "We're  hoping  they'll  be   intrinsically  less  toxic,"  says  Marsh.  

Used cooking oil could be turned into cosmetics New Scientist 05 March 2007 Would you consider rubbing used oil from a deep-fat fryer into your face? Oil from restaurant kitchens may soon be recycled into products ranging from cosmetics and soaps to industrial surfactants used to clean up oil spills. Researchers at Dowling College in Oakdale, New York, found that used vegetable oil can be fermented with the yeast Candida bombicola to produce "biosurfactants". Surfactants play a key role in products as diverse as detergents and pesticides. The yeast process creates biodegradable surfactants called sophorolipids, which are often used in skin and hair products. Normally, though, these surfactants are made from petroleum, are not biodegradable and can interfere with the life cycles of some aquatic organisms. Waste cooking oil is a great untapped resource, says lead researcher Vishal Shah. Restaurants and hotels in the US alone produce more than 11 billion litres of the stuff each year, the majority of which is disposed of as sewage or in landfill sites. "We are turning an environmentally harmful waste product into something that is both safe and useful," says Shah. The prospect of turning waste cooking oil into biosurfactants has been investigated before, but Shah and colleagues have demonstrated the feasibility of the process by showing it can be done cheaply in commercial fermenters (Biotechnology Progress, DOI: 10.1021/bp0602909). Biosurfactants could net as much as $20 per litre when used in cosmetics, says Shah.

New Scientist | 17 February 2007 Editorial: Self Defence Over Bird Flu is No Crime Good for Indonesia. There, it's been said. The country at the centre of the H5N1 bird flu storm has stopped sending virus samples to the World Health Organization. Though this means that scientists cannot track H5N1's increasingly worrying evolution, which is bad, Indonesia is doing the only thing it can to protect its people. It has also brought an unpalatable truth out into the open. In a fair world, Indonesia would send its virus to the best labs and share in any vaccine made from it. In our world, Indonesia sends off its virus, companies make vaccine from it and sell it to countries that can pay. Indonesia is not one of them, and neither are the other countries suffering badly from H5N1. Indonesia is treating this as a case of biopiracy. Like other tropical countries, it is a hotspot of biodiversity. For decades foreign companies have helped themselves to its plants, microbes or whatever, and made lucrative products from them. In response, it has passed laws to stop exports of genetic material without agreement. It is invoking such a law to control samples of H5N1 and asking developed countries to respect the sort of intellectual property rules that they themselves have imposed for decades. It is not just looking for money. Everyone in the vaccine business knows that when a pandemic appears, countries with vaccine factories will ensure their own citizens are catered for before any vaccine gets exported. That is what happened in the swine flu scare in the US in 1976. If a pandemic does begin, countries without factories will probably not receive vaccine in time for the first wave. By withholding the virus, Indonesia is leveraging the one resource it has to obtain flu vaccine, possibly even its own factory. As Lily Sulistyowati of Indonesia's health ministry put it: "Indonesia's state-owned drug maker Bio Farma does not have the technology and expertise to create the vaccine. We can only offer foreign pharmaceutical companies our strain of the virus." The country says it is will do this only under material transfer agreements that ban commercial use except by prior agreement. It is a shame things have come to this. Indonesia should be able to share its virus freely without feeling that it is sacrificing the one chance it has to save its people from a pandemic. Perhaps vaccine manufacture should be a global public good, not a national or private one -- something this magazine has noted before. We need a system that works for everyone. In its absence, those material transfer agreements should be signed now. We need to see what H5N1 is up to in Indonesia. From issue 2591 of New Scientist magazine, 17 February 2007, page 3

New Scientist | 26 November 2007 Indonesia fights for rights to bird flu samples Indonesia will not share bird flu virus samples unless richer countries agree to give developing nations control over their use and access to cheap vaccines, a spokeswoman from the nation's health ministry said on Monday. Health officials from around the world failed to reach an agreement on a new virus sharing system at talks hosted by the World Health Organisation (WHO) in Geneva last week. Indonesia, the nation worst hit by bird flu with 91 human deaths, has held back its virus samples since August 2007 and wants guarantees from richer nations and drug makers that poor countries get access to affordable vaccines derived from the samples. Health Minister Siti Fadillah Supari insisted on "equitable sharing of benefits arising from the use of viruses" at the meeting. Sharing samples is deemed vital to determining whether the viruses are mutating, becoming drug resistant or growing more transmissible. "Talks hit a deadlock because the health minister was relentless in pushing for a 'material transfer agreement' for each virus sample, but not everyone agreed to that," ministry spokeswoman Lily Sulistiowati says. "We hope that negotiations will continue." Intellectual property Indonesia wants a material transfer agreement for each virus sample sent to foreign labs, specifying that the sample will only be used for diagnostic purposes and not for commercial gain. Under this proposal, any commercial use of the virus would require prior consent of the country providing it. By retaining the intellectual property rights, Indonesian officials say, a country could allow access to global vaccine stockpiles at an affordable price. The WHO agreed last May to revamp its 50-year-old system for sharing flu virus samples with researchers and drug firms. It had wanted its 191 member states to adopt an agreement by May, but divisions remain. Experts fear the constantly mutating H5N1 virus could change into a form easily transmissible among humans and sweep the world, killing many millions of people worldwide. Voluntary donations According to the WHO, sharing samples is vital for tracking the deadly H5N1 virus and developing vaccines against a potential pandemic. Jakarta has shared just two specimens this year, both from Indonesian women who died in the popular tourist resort of Bali in August, according to the WHO. Sixteen companies are at various stages of licensing a vaccine for H5N1. These include GlaxoSmithKline, which announced last June it would donate 50 million doses of its "prepandemic" bird flu vaccine to WHO's global stockpile. The Indonesian government and a unit of the US firm Baxter International have also agreed to develop a vaccine. Under the accord, Jakarta has been supplying virus specimens while Baxter is providing technology to develop the vaccine.