Update of Concerns #2 (22.9.99)
Written and compiled by Mae-Wan Ho and Angela Ryan
1. African group tables important proposal at the World Trade Organisation which may revoke and ban biopatents. The full proposal can be found at the WTO website.
The African group questions the TRIPS requirement for mandatory patenting of some life forms and some natural processes. They propose that all plants, animals and micro-organisms should not be patentable. They seek clarification that "sui generis" system of plant varieties protection can include systems that protect the intellectual rights of indigenous and farming communities. They have also asked for TRIPS to be made to harmonize with the Biodiversity Convention and the FAO's International Undertaking on Plant Genetic Resources.
This proposal is most significant. ISIS have given it our wholehearted support, as it may lead to international agreement that all biopatents will be banned.
Please support the African Group's position by sending your name, organization and address to email@example.com , so that the issue can be made high priority for the WTO Seattle Conference December 2-3, 1999.
2. Anglo-American Agreement to protect Human Genes
British Prime Minister Tony Blair has initiated an extraordinary deal with US President Bill Clinton to negotiate an Anglo-American agreement to protect the 100,000 genes of the human body. The deal aims to prevent entrepreneurs profiting from gene patents and to ensure that the benefits of research are freely available world wide to combat or even eliminate diseases.
The two leaders aim to ensure that the world's largest medical charity, the British-based charity, Wellcome Trust, and the US government's National Institute of Health, publicize gene-sequences within 24 hours of their discovery - so that the benefits accrue entirely to the public. It is thought that research institutions, universities or laboratories would be obliged to waive their rights to patent their work in the public interest.
ISIS welcomes this move by Blair and Clinton. The next step is to prevent all living organisms, seeds, genes and cell lines from being patentable.
B. New Evidence of Hazards and Potential New Hazards
1. The human mouth and pharyngx contain bacteria that can take up and express transgenic DNA, including antibiotic resistance marker genes. This confirms the ability of transgenic DNA to spread by horizontal gene transfer.
The findings: A genetically engineered plasmid was found to survive (6 to 25%) up to 60 min. of exposure to human saliva. Partially degraded plasmid DNA was capable of transforming Streptococcus gordonii, one of the bacteria that normally live in the human mouth and pharynx and are naturally transformable. The frequency of tranformation dropped exponentially with time of exposure to saliva, but it was still greater than 10-7 after 10 mins. Transformations were also obtained when plasmid DNA was mixed directly with filter-sterilised human saliva. Approximately 107 transformants were obtained per microgram plasmid DNA. Whole (unfiltered) saliva had a background of erthyromycin-resistant bacteria, but transformation of the test strain occurred nevertheless at a high frequency of 10-4 of all colonies of S. gordonii isolated. Human saliva contains factors that promote competence of resident bacteria to become transformed by 'free' or 'naked' DNA.
Our comment: Transgenic DNA from food is unlikely to be completely broken down in the mouth, and may transform bacteria normally present in the mouth. One main danger is the uptake of transgenic DNA containing antibiotic resistance marker genes by the bacteria, but other genes and novel constructs involving viral promoters/enhancers may also be hazardous.
Reference: Mercer, D.K., Scott, K.P., Bruce-Johnson, W.A. Glover, L.A. and Flint, H.J. (1999). Fate of free DNA and transformation of the oral bacterium Streptococcus gordonii DL1 by plasmid DNA in human saliva. Applied and Environmental Microbiology 65, 6-10.
2. Special Report comissioned by UK Ministry of Agriculture, Fisheries and Food (UK MAFF) shows that DNA is not readily degraded by most commercial processing procedures. Animal feeds are likely to contain gene-size DNA fragments. One main conclusion is that there is significant risk of transmitting transgenes in the gut of farm animals, and recommends against using ensilaged GM crops as animal feed.
The findings: Samples of oil seed, linseeds, soya and wheat were analysed for DNA fragments after various treatments. Grinding and milling left DNA largely intact, as did treatment with dry heat for 30 mins. at 90 deg. C. Degradation to less than 100bp occurred only after treatment with dry heat at 96 deg.C or moist heat at 93 deg. C. DNA was completely stable in silage.
Reference: Forbes, J.M., Blair, D.E., Chiter, A. and Perks, A. (1998). Effect of Feed Processing Conditions on DNA Fragmentation Section 5 - Scientific Report, UK Ministry of Agriculture Fisheries and Food, London. Our Comment: It should not be assumed that processed food contains no DNA. The degradation of DNA is defined as less than 100bp. We now know that (see below) nucleic acids as small as 25bp can stimulate autoimmune reactions.
3. Gene therapy and naked DNA vaccines can trigger autoimmune reactions. New research shows that any fragment of double-stranded (ds) DNA or RNA introduced into cells can induce these reactions. (Thanks to Brian Goodwin for drawing our attention to this item.)
The findings: Immune reactions are normally mounted by white blood cells against substances (antigens) foreign to the body, and involve the expression of many different genes. Autoimmune diseases are associated with abnormal reactions of other cells directed against the body's own constituents, and can be triggered by viral infections. Autoimmune diseases can be specific to organs or cells, examples are rheumatoid arthritis, insulin-dependent diabetes and Graves disease of the thyroid. The researchers found that introducing any fragment of dsDNA or dsRNA into the cells by transfection stimulate the abnormal expression of major histocompatibility complex (MHC) class I and class II genes as well as other genes involved in presenting antigens on the surface of the cell membrane. This in turn induces activation of immune cells against the antigen-presenting cells. The effects were indifferent to the sequence of nucleic acid introduced, so long as it is double-stranded; and fragments as short as 25 base pairs were effective. The authors conclude, "This phenomenon may contribute to the development of autoimmunity when plasmid DNA is introduced during gene therapy and may be important when dsDNA is used in plasmid DNA vaccinations."
Reference: Suzuki, K., Mori, A., Ishii, K.J., Singer, D.S., Klinman, D.M., Krause, P.R. and Kohn, L.D. (1999). Activation of target-tissue immune-recognition molecules by double-stranded polynucleotides. Proc. Natl. Acad. Sci. USA 96, 2285-90. Our comment: This research not only raises serious safety concerns over gene therapies and the use of naked DNA vaccines, it also emphasises the need to regulate the ever-increasing use of all kinds of naked DNA and RNA such as plasmids and vectors and to prevent their discharge into the environment. In view of the fact that fragments as small as 25bp can stimulate autoimmune reactions, naked DNA or RNA should be fully degraded before being discharged.
4. A scientist from the Center for Complex Infectious Diseases in Rosemead, California, claims to have found a new virus associated with chronic fatique syndromes which is part bacteria. Could genetic engineering have contributed to creating it?
The findings: More than 50 bacterial genes were found in a virus isolated from patients with various chronic fatique syndromes. The scientist regards this as a new organism and coined the term, "viteria" to describe the hybrid virus-bacteria. The virus most closely resembles a cytomegalovirus. And top of the list of bacteria from which the virus has captured genes are E. coli and Bacillus subtilis. Our comment: This new discovery should be seen in the light of a report on the possible links between genetic engineering biotechnology and the resurgence of infectious diseases co-authored by seven scientists, four of whom have signed onto the World Scientists' Statement (Ho, M.W., Traavik, T., Olsvik, R., Tappeser, B., Howard, V., von Weizsacker, C. and McGavin, G. (1998). Gene technology and gene ecology of infectious diseases. Microbial Ecology in Health and Disease 10, 33-59.) The report was among the top ten 'Project Censored' stories in 1998. It drew attention to, among other things, pathogenicity islands coding for many virulence genes that transfer horizontally as a unit, so non-pathogens can be converted into pathogens in a single step.These pathogenicity islands are thought to have originated from bacterial viruses which have integrated into bacterial genomes and picked up a range of virulence genes. They are highly mosaic, consisting of parts of plasmids and bacteria, particularly of those parts that might have been assembled into artificial vectors for genetic manipulation. Could genetic engineering have inadvertently contributed to creating pathogenicity islands? Could the newly discovered 'viteria' be pathogenicity islands that have gained independent existence as infectious particles? These are questions which urgently need to be addressed. Cytomegalovirus, which the viteria most resembles, is one of the first viruses to be used as vector in genetic manipulation of animals, while E. coli and B. subtilis, the top two bacteria from which it has acquired genes are also the two most commonly used bacteria in genetic engineering.
Reference: Martin, W.J. (1999). Bacteria-related sequences in a simian cytomegalovirus-derived stealth virus culture. Experimental and Molecular Pathology 66, 8-14.
5. A new genetic engineering technique, 'chimeroplasty' claims to overcome the current hit or miss transgenic technology that results in random gene insertions and rearrangements; instead, it can change a single base at a predetermined position in a specific gene in the plant cell. But it may not be as precise as claimed and introduces new dangers. (Thanks to Suzanne Wuerthiele for drawing our attention to this paper.)
The findings: A technique of directed gene conversion involves introducing a palindromic inverted repeat of a sequence of 25 bases composed of DNA and modified RNA residues, which forms a stable hairpin. The sequence is homologous to that of the target gene, except for the base to be substituted, which is in the middle of the sequence of 25 bases. When introduced into the cells by a gun that shoots tiny gold particles coated with the hairpins, the hairpins will basepair with both strands of the target sequence in the gene. DNA mismatch repair enzymes will then convert the sequence of the gene to that specified by the hairpin. Using this technique, the researchers attempted to convert a gene in tobacco coding for the enzyme acetolactate synthase to a herbicide-resistance phenotype, by changing the codon (CCA) for proline at amino-acid position 196 to CAA for glutamine and CTA for leucine respectively, with two different hairpin constructs. The results show that the target sequence was converted, but not at the base intended. Conversions were at neighbouring bases. For example, ACA for threonine was obtained instead of CAA intended, and TCA for serine resulted, as well as ACA and TCA instead of the intended CTA. Another complication is that the gene is capable of undergoing spontaneous mutations to herbicide resistance. The directed mutation rates were up to 20-fold those in controls, but were variable from experiment to experiment.
Our comment: It is very unlikely that the technique is as precise as claimed. The mere act of introducing nucleic acid sequences into the cells by bombardment with a particle-gun will trigger injury responses that can cause nonspecific recombination. In addition, the technique depends on imprecise basepairing between the target sequence and the introduced hairpin. Can one be sure that nontarget sequences are never affected? The hairpins themselves are a hazard to biodiversity and health if released into the environment. All kinds of unintended gene conversions could take place in species exposed to the constructs, including human beings.
Reference: Beetham, P.R., Kipp, P.B., Sawycky, X.L., Arntzen, C.J. and May, G.D. (1999). A tool for functional plant genomics: Chimeric RNA/DNA oligonucleotides cause in vivo gene-specific mutations. PNAS 96, 8774-8778.
6. "The Herbicide, Glufosinate, used with millions of acres of GM crops including corn, canola and soy, causes birth defects on exposure of father alone as well as mother!" submitted by Joe Cummins. Joe Cummins has written a number of previous notes on the danger of the herbicide ,glufosinate, used with GM and normal crops and on the false claims by officials of EU , US and Canada that the herbicide has no harmful side effects. The previous evidence showed that pregnant females fed food containing the herbicide gave birth to children with birth defects, as well as defects in behavior and learning. Learning defects were also experienced by young children exposed to the herbicide. Recent studies showed that fathers exposed to glufosinate gave birth to children with birth defects while most other pesticides did not produce the same effect.
Reference: Garcia,A.,Benavides,F.,Fletcher,T. and Orts,E. (1998). Paternal exposure to pesticides and congenital malformations. Scand J Work Environ Health 24, 473-80.
Joe Cummins comments: The glufosinate birth defects suggest that the large chemical companies have undue influence over government bureaucrats . Such bureaucrats turn their backs on clear evidence of danger from pesticides and promote dangerous genetic engineering.
7. A new study looks at the prevalence and spatial distribution of viruses in natural populations and discusses the implications of widespread multiple viral infections in natural plant population with respect to the release of transgenic plants expressing virus-derived genes. A quote from this paper: " The presence of transgenic virus resistant plants expressing viral proteins or virus-derived nucleic acids introduces a substantially new dimension into the dynamics of plant-virus co-evolution, even though virus-derived nucleic acids are normal constituents of plant populations. There is a possibility that the spread of virus-derived transgenes through seed and pollen will substantially alter the distribution of viral nucleic acids, for example, gene flow might reduce temporal and spatial variation in the incidence of virus-derived nucleic acids and so increase the potential for recombination."
Reference: Raybould et al (1999) The prevalence and spatial distribution of viruses in natural population of Brassica oleracea . New Phytol 141, 265 275
6. "Virus-Resistant Crops Could Help Weeds" Says Professor Alison Power. Genetic engineering cereals to resist the barley yellow dwarf virus (BYDV) might indirectly cause farmers difficulties in controlling related weeds. A report presented at the Ecological Society of Americas' annual meeting indicates that the resistance engineered into oats could spread to wild oats, a weed. Transgenic barley and oats that can resist BYDV have been developed, but there is concern that because these crops can hybridize with wild relatives, that the introduced genes will escape into related weeds. Alison Power, an ecologist at Cornell University says that if wild oats gain resistance to BYDV, they could become a much larger problem for farmers, and might also disrupt natural habitats, outcompeting other native species. Power grew oats and wild oats in greenhouses and infected them with the BYDV. She found that infected wild oats did not perform well: they were much thinner and had shorter roots than uninfected controls and infected oats. Infected wild oats also produced fewer seeds than normal. "A BYDV-resistant transgene transfer seems likely to help wild oat survivability," concludes Power.
Reference: Contact: Alison Power, Department of Ecology and Evolutionary Biology,E331A Corson Hall, Cornell University, Ithaca, NY 14853,USA.
7. An independent study conducted by Dr Marc lappe at CETOS, has shown that the phytoestrogen concentrations in two varieties of GM herbicide tolerant soyabeans were reduced by an average of 12-14 % compared to their non GM counterparts. Most of the reduction was attributable to reductions in genistin and to a lesser extent diadzin levels, which were significantly lower in modified compared to conventional soybeans in both strains. (Thanks to Dr Michael Antoniou for forwarding this paper) Soyabean-based food products are of growing medical interest as they contain two key biologically active ingredients, notably genistin and diadzin, which are phytoestrogens. A number of studies have indicated that phytoestrogen content of ingested soybeans can modify the pathogenesis of some hormone-dependent and hormone-independent diseases and may constitute a natural estrogen replacement therapy in post menopausal women. In the United States approximately one half of the 1998 soyabean crop consisted of GM, herbicide-tolerant soyabeans, of which 33% are slated for export, thus the phytoestrogen content of GM soyabeans is of international interest. Given the high biological potency of isoflavones and their metabolic conversion products, these data suggest genetically modified soybeans may be less potent sources of clinically relevant phytoestrogens than their conventional precursors. In order to ensure uniformity of clinical results the scientists who conducted this study strongly suggest there is a need to establish baselines of expected isoflavone levels in transgenic and conventional soy products so as such data will be available when making clinical decisions.
Reference: Lappe et al (1999) Alterations in Clinically Important Phytoestrogens in Genetically Modified, Herbicide-Tolerant Soybeans. Journal of Medicinal Food, Vol 1, no 4.
Our comment: The unpredictability in the composition of GM crop plants is highlighted by this study and further confirms the inadequacy of the regulatory protocols, which rely on the principle of 'substantial equivalence' for approving GM crops and products.
8. An investigative safety assessment study carried out at the Scottish Crop Research Institute in Dundee (completed in 1997) has shown that Agrobacterium used in the production of GMOs is a possible vehicle for gene escape, or horizontal gene transfer. The findings: The study shows that the use of frequently used antibiotics, carbenicillin, ticaracillin and cefotaxime failed to eliminate contamination of Agrobacterium even after 13 months of repeated subculture from transformed and weaned plants. Furthermore a significant percentage (12.5%) of Agrobacterium containing the binary vector was detected in tissue culture after six months, even without selection pressure being maintained. The MAFF, R&D Surveillance report No. 395 states that: " The presence (contamination with) of disarmed (but foreign gene containing) Agrobacterium in GMO plant tissues would not represent a risk if the binary vector had been 'lost', but the discovery that it survives, even without selection advantage, indicates that gene spread is a real possibility."
The presence of latent Agrobacterium was also observed during the tissue culture phase of GMO production. The scientists who conducted this study stress the need for sampling tissues for contamination and not merely relying on the lack of visual symptoms. They strongly suggest that such procedure should be adopted routinely, for in the case of Agrobacterium, sampling is the only reliable method of indicating contamination.
References: McNicole et al (1997) The Possibility of Agrobacterium as a Vehicle for Gene Escape. MAFF. R&D and Surveillance Report: 395. Carol Barrett et al (1997). A risk assessment study of plant genetic transformation using Agrobacterium and implications for analysis of trangenic plants. Plant Cell. Tissue and Organ Culture 47: 135-144. Our Comment: It is claimed that other more effective antibiotics are now used to eliminate Agrobacterium from GM plant material and the above mentioned sampling procedures are now thought to be implemented. But, before the results of the above study became available, it is likely that most GM plants transformed by Agrobacterium were contaminated with residual Agrobacterium containing antibiotic resistance genes. Most of the present generation of GM crops commercially grown or being field-tested were, in fact, created before the results of this study were published and therefore carry this risk. The above study demonstrates that major flaws in safety assessments are being uncovered only after the products have been approved.
9. A new study on the effects of a genetically engineered microorganism (GEM) on soil biota and plant growth has shown that GEMs can persist under conditions found in some soil ecosystems for long enough periods of time to stimulate major changes in soil biota that could affect nutrient cycling processes. Nematode community composition and plant growth were also affected following the introduction of the GEM.
Reference: Holmes M T et al (1999). Effects of Klebsiella planticola SDF20 on soil biota and wheat growth in sandy soil. Applied Soil Ecology 11 (1999) 67-78. Our comment: Further investigations are needed to determine the long term effects of these observations and whether the release of GEMs have a viable future. The potential for an ecological effect to occur after the release of GEMs is now apparent and of global concern.
And now the goood news...
10. Non-functional genes in patients with Fanconi anaemia undergo corrective mutations spontaneously to restore normal function. These are the latest examples of functional corrections that can occur in a range of different hereditary defects which show how fluid and dynamic genes and genomes are. Molecular geneticists should investigate the physiological and environmental factors favouring such functional corrections of defective genes instead of concentrating exclusively on changing and transferring genes. (Thanks to Wytz de Lange for drawing attention to this paper.) The findings: Somatic cells in individuals with non-functional pathogenic alleles (forms of a gene) have recently been discovered to revert spontaneously to functional, wild-type alleles. The best known mechanisms occur in heterozygous patients, ie, patients with two different alleles of the gene and involves recombination within the gene during ordinary cell division, in which the two alleles present exchange parts, or gene conversion, in which the functional allele converts the non-functional. New mechanisms of corrections are described for non-functional alleles in homozygous patients (those with two identical alleles). Frameshift alleles in two different patients were restored to the correct reading frame, in one case, by deletions of two single base-pairs, and in the second, by insertion of 5 base-pairs. A missense mutation in another patient (T to G in position 1749) was compensated by a C to T mutation in position 1748. Although the protein in all three cases differed in amino-acid sequence from the wild-type, they were functionally equivalent to the wild-type.
Our comment: These are the latest examples of the fluidity of genes and genomes which has completely invalidated the genetic determinist science driving and promoting genetic engineering biotechnology. Evidence accumulating since the mid 1970s indicates that there is nothing constant about genes and genomes.The new observations that even nonfunctional genes can revert to functional wild-type or wild-type equivalents take genomic fluidity one step further. Rather than concentrating exclusively on changing and transferring genes, molecular geneticists ought to direct their attention to the physiological and environmental factors that favour the regulation of gene function and structure in the whole organism, even to the extent of correcting the structure of non-functional genes.
Reference: Waisfisz, Q., Morgan, N.V., Savino, M., et al (1999). Spontaneous functional correction of homozygous Fanconi anaemia alleles reveals novel mechanistic basis for reverse mosaicism. Nature Genetics 22, 379-383
C. New Postings on ISIS website: www.i-sis.dircon.co.uk
1. Draft Open Letter to all governments from world scientists to be presented to WTO conference in Seattle, Washington, December 2-3, 1999. It re-iterates our World Scientists' Statement call for a 5 year moratorium and a ban on biopatents.
We are encouraging other scientists to sign on and non-Government organisations to endorse our Statement and Letter. Either sign onto Statement on our website, or e-mail us at firstname.lastname@example.org
2. From BSE to GMO - What Have We Learned? by Dr. Harash Narang, with Introduction by Angela Ryan. This is a new ISIS publication in collaboration with the Millenium Debate, due to be released 26 September, 1999.
3. The precautionary principle by Dr. Vyvyan Howard and Prof. Peter Saunders, Published in Nature correspondence, 16 September, 1999.
4. Head to Head, by Mae-Wan Ho, to appear in Sovereign magazine.
5. No to GMO, Civil Society versus Corporate Empire, talk presented in Progressive Farm Leaders Summit on Genetic Manipulation and Agriculture, Coalition of Family Farmers, USA, Mannassas, Virginia, September 11, 1999.