Report LIFE 98

Conclusions and recommendations

Overview of the process

Because the ambitions formulated in our proposal where rather high, including the collaboration of several laboratories, completion of the project has taken longer than originally planned. In addition, it became necessary to adapt particular strategies in order to attain the goals that were set. In spite of the complexity of the process the major part of these goals have been attained.

Results for potential targetgroups

Potential target groups are governmental organisations, universities, or companies (household, commercial, industrial, and small community water treatment industry) involved in water quality monitoring programmes (and in the future possibly air and food monitoring).

Any pro-active company that is concerned with sustainable development should in the near future be able to test e.g. their core chemicals or effluents with our in vitro test systems. Foreseeing possible regulations on the European level this should apply to any company discharging effluent in the environment. Other potential target groups are chemical companies that develop raw materials for synthesis, plasticizers, stabilizers or pharmaceutical companies that develop pesticides or medication. The test should make an easy screening of their pure compounds for their binding capacity on the estrogen receptor possible.

Filtering of water and the regular consumption of appropriate health food target at the entire population, including household use of drinking water and all individual users. These target groups all over the world should benefit from the developments elaborated by this project in terms of decreased exposure to hormone disrupting agents, with resulting decreased risk of diseases such as male infertility, particular congenital abnormalities, and particular types of cancer.

Comparison to the project-objectives

The vast majority of the project-objectives have been attained, albeit using technology that is not always identical to the one proposed in the original proposal.

The novel methods for the detection of hormone disrupting agents which exert their deleterious effects after binding to the human estrogen-receptor has been validated and tested on real-life environmental samples. A new approach has been taken to the technology for the elimination of hormone disrupting substances from water. The efficacy of the health-food principle for the reduction of uptake of xeno-oestrogens from ingested food has been documented in a simulator of the digestive tract.

Project’s Benefits

The problems of decreased reproductive health in the male and increased prevalence of breast cancer in the female are encountered world-wide. They were ascertained by several studies in many of the European members, from Greece to Denmark. It is difficult to estimate the economic cost of the decline in fertility, the rise in certain cancers or other endocrine disruptions in animals or man. In a worst case scenario this could eventually lead to the extinction of species and the loss of balance in nature. In 1991 scientists hypothesised that hormone disruption could be the cause of declines in the populations of some wildlife species which have occurred over the past 50 years.

Without any doubt, reproduction is one of the most fundamental processes in humans and other vertebrates. The increasing prevalence of infertility in man is illustrated by the steady rise of couples in Europe undergoing medical assisted conception during the last decade. If we only consider the high cost of treatment of infertility which can amount up to 800.000 BEF per birth after certain techniques of assisted reproduction, it becomes obvious that the advantage of preventing infertility is enormous. Apart from the high cost, fertility treatment involves serious physical and mental stress whose financial impact is even more difficult to estimate. Furthermore, certain techniques of assisted reproduction, which are needed to solve infertility, are potentially dangerous to the mother and the offspring (e.g. genetic defects).

From an environmental viewpoint, our project aimed at decreasing the exposure of men to endocrine disrupting compounds by reducing the pre-intake from water thanks to a filter system, and the post-intake adsorption from the intestine thanks to a Lactobacillus based "Health-Food". Moreover important steps were taken in the development of a complementary set of in vitro bioassays addressing the current international need for in vitro assays with different end point detection. The Sertoli cell assay with its high relevance for the male (the only specific assay in this regard available) and the hypothalamo-pituitary (brain) assay will substantially help to reduce the risk of overlooking hormone disrupting substances in the rush to screen chemicals for estrogenic effects.

In the long run, the project will contribute substantially to the knowledge on the mechanisms of action of those compounds on the endocrine system, and on the dispersion of endocrine disrupters in the environment, leading to recommendations for regulatory measures at national, European Union or world-wide level. These recommendations should result in a safer and cleaner environment and should consequently lead to an improvement of the life quality (physical and mental) for all living creatures

From a an industrial viewpoint it is clear that the elimination of the compounds of concern in surface or in drinking waters would require a highly specific and very expensive technology because of the low concentrations and the low biodegradability of the chemicals. The catalytic properties of the manganese particles in symbiosis with manganese regenerating microorganisms could offer a simple, cost effective solution to this.

Furthermore, research into different modes of actions of endocrine disrupters, leading to the possible identification of new hormone disrupters, can result in certain regulatory measures (on a national, European Union or world-wide level) bridling the use and production of particular chemicals in industrial processes or for agricultural or household use. We refer to the Paris Commission Recommendation (92/8) concerning the phasing out of certain alkylphenols/ ethoxylates (AP and APE’s) as cleaning agents for industrial and domestic use by the year 2000. Obviously this has a highly economical relevance for the industry.

We are convinced that the technology used for the screening of endocrine disrupters will be applicable in any laboratory world-wide disposing of basic analytical or cell culture equipment. As to the development of the filter system and the Lactobacillus "Health Food", these should eventually lead to mass production and to a distribution via normal commercial channels, making them attainable for any person in the European Union

From experience in the past it has become evident that the interest for the field of human fertility is enormous, and that any development in this area meets with a wide response. To substantiate this, we refer to the broad public impact of broadcasts by the BBC and Belgian television stations on the decline of human fertility, and to the success of the book entitled "Our Stolen Future" by Colborn et al.

Application possibilities in other branches

We are evaluating the use of the developed assays for the measurement of the total estrogen load in the blood of patients and investigate if this could be linked to different pathologies:

• assessment of the total estrogen load in relation to pathologies such as male infertility, breast and prostate cancers.
• Total estrogen determination linked to osteoporosis in the male.

Furthermore, certainly when applied to the beta estrogen receptor this experimental set-up could be an important tool in the search for selective alfa receptor or beta receptor binders, which is extremely important in menopausal replacement treatment and cancer prevention or treatment. The tests are now being evaluated for the screening of the (alfa) estrogenic effects of fytoestrogens present in beer.

The last couple of years, important scientific contributions in the field of endocrinology have revealed a clear cell type and promotor context specificity of estrogen response. This is the experimental paradigm behind the booming research into selective estrogen modulators (SERMs) used for the treatment of osteoporosis, breast cancer and the post-menopausal syndrome. Our tests will prove to be important tools for the study of the fertility related effects of these compounds.

The novel, cell-free test system for estrogenic substances will find its application in studies regarding the link between the environment and population health, and it will provide a rapid and accurate tool for assessing the possible side effects of certain industrial developments.

Innovative character of the project

The creation of a novel essay for the detection and quantification of estrogen-like substances, that does not make use of living cells, is innovative and signifies a real break-through in this field. Also, the concept of systems for the elimination of hormone disrupting agents from surface and/or drinking waters, and for the inhibition of uptake of such agents from ingested food are innovative.

Effectiveness of dissemination activities

A high level of dissemination of the results of the present project has been attained thanks to multiple presentations in scientific papers and congresses, as well through communication of particular aspects in the lay press. Awareness has been created for the problems of environmental contamination with hormone disrupters, and this has stimulated the implementation of regional, national and -most of all- international regulations aiming at eliminating these agents.

Potential

Herebelow, excerpts are given of a study of the functional food market in Europe by the Leatherhead Research Association. It is illustrative of how the rising concern of people with their health status can result in important economic revenue. It is clear that all the techniques that have been developed in this project will aid in the reduction of noxious agents in the environment, and thus in an improvement of the general health status of men.

"With the current focus on disease prevention and the quest for optimal health, the use of probiotic market potential is enormous. Today the two major categories for probiotic products are functional foods, such as yogurt enriched with active cultures, primarily L. acidophilus, and probiotics in dietary supplement form. The functional food market in the UK, Germany and France equal more than 1 billion euro a year when products were limited to those making specific health claims on the packaging or in advertising. The German market is largest, followed by France and UK (Leatherhead Food RA, quoted in Hillam, 1999). Products consist of probiotic yogurts, fermented milk drinks, dental health chewing gums, high fibre breakfast cereals, oat-based breakfast cereals and dietetic biscuits. Hilliam's earlier report of the Leatherhead Food RA data illustrates growth in the European functional food market, even in just two years. In 1997, the market for functional foods in the UK, France, Germany, Spain, Belgium, the Netherlands, Denmark, Finland and Sweden was worth approximately euro 1.37 billion. Dairy products formed about 65% of the market based on the popularity of probiotic yoghurts. Probiotic yoghurts had a value of euro 825 million in 1997 and represented 10% of all yoghurts sold.

Industry respondents predicted strong growth in the functional foods industry to 2003. Almost half felt that functional foods would account for 5-10% of total European retail food and drink sales within this period.

Reuters Business Insight (1999), predicted that by the year 2002, functional dairy products will account for 45% of nutraceutical sales in Northern Europe."

Relevance to the EU legislative framework

The results of our project bear major relevance to the European legislation, since the novel method for the detection of estrogen-like hormone disrupting substances renders adequate mapping of environmental pollution possible, and should help inducing regulatory legislation aiming at the elimination of the sources of such pollution. In addition, monitoring of the effects of these legislations on the environment has become feasible by the novel technology, and the health effects of implementing new methods and criteria for water filtration as well as health food consumption on the prevalence of certain diseases in Europe are part of the assignment of the European governing institutions.