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IFST updated Information Statement on Phytosterol Esters (Plant Sterol and Stanol Esters)

21.01.2005


The Institute of Food Science & Technology, through its Public Affairs and Technical & Legislative Committees, has authorised the following Information Statement, dated January 2005. This cancels and replaces that dated June 2000.



Summary

The term "phytosterols" covers plant sterols and plant stanols. Plant sterols are naturally occurring substances present in the diet principally as minor components of vegetable oils. Plant stanols, occurring in nature at a lower level, are hydrogenation compounds of the respective plant sterols. An elevated level of blood cholesterol is one of the well established risk factors for coronary heart disease. Both plant sterols and plant stanols are effective in lowering plasma total and low density lipoprotein (LDL) cholesterol and this occurs by inhibiting the absorption of cholesterol from the small intestine. In order to achieve a cholesterol lowering benefit approximately 1g/day of plant sterols or plant stanols need to be consumed. In comparison, normal dietary intake is between 200-400mg/day plant sterols whereas the normal dietary intake of plant stanols is negligible.


The enrichment of foods such as margarines with plant sterol ester or plant stanol ester is one of the recent developments in functional foods to enhance the cholesterol lowering ability of traditional food products. The consumption of average amounts of spread per day (approximately 20g in Western Europe) supplemented with between 8-10% plant sterol or plant stanol lowers serum total cholesterol and LDL cholesterol by 8-13% - this is equivalent to the consumption of between 1.6 - 2.0g plant sterols or plant stanols per day. Plant sterols and plant stanols appear to be without hazard to health, having been shown without adverse effects in a large number of human studies. They show no evidence of toxicity even at high dose levels and gastro-intestinal absorption is low. In the United States a panel of independent experts has concluded that vegetable oil plant sterol esters and plant stanol esters are safe for use as an ingredient in vegetable oil spreads. Furthermore in the European Union an opinion from the Scientific Committee on Food (SCF) concluded that the use of phytosterol-esters in yellow fat spreads (maximum level of 8% free phytosterols) is safe for human use. More recently the SCF and its successor, the European Food Safety Authority (EFSA) through its Scientific Panel on Dietetic Products, Nutrition and Allergies, have recommended that sterol-containing foodstuffs should not be consumed in amounts resulting in total phytosterol intakes exceeding 3 g/day.

Introduction

Phytosterols (including plant sterols and stanols) are natural components of edible vegetable oils such as sunflower seed oil and, as such are natural constituents of the human diet. The plasma cholesterol-lowering properties of plant sterols have been known since the 1950s (Pollak, 1953). Early studies examined the cholesterol-reducing potential of plant sterols using up to 25g/day consumed in solid crystalline form. Over the years however, it was established that sterols dissolved in edible fat products are more effective at reducing blood cholesterol levels than sterols in crystalline form. It is difficult to incorporate free sterols into edible fats/oils because of their insolubility, whereas sterols esterified to fatty acids are more fat soluble. In the intestine, most sterol esters are hydrolysed to free sterols as part of the normal digestive process.

Plant stanols are the hydrogenated counterparts of the plant sterols but are less abundant in nature than the corresponding plant sterols. Consequently, the normal dietary intake of plant stanols is much less than that of plant sterols. An elevated level of blood cholesterol is one of the well established risk factors for coronary heart disease. Blood cholesterol levels can be decreased by following diets which are low in saturated fat, high in polyunsaturated fat and low in cholesterol. Although considerable achievements have been made in terms of knowledge and education, consumers still find it difficult to follow healthy eating advice. The enrichment of foods such as margarines with plant sterols and stanols is one of the recent developments in functional foods to enhance the cholesterol-lowering ability of traditional food products.

What are plant sterols and stanols?

Plant sterols have a role in plants similar to that of cholesterol in mammals, e.g. forming cell membrane structures. Plant sterols fall into one of three categories: 4-desmethylsterols (no methyl groups); 4-monomethylsterols (one methyl group) and 4,4-dimethylsterols (two methyl groups). The most common plant sterols are β-sitosterol, campesterol and stigmasterol and structurally these are very similar to cholesterol, belonging to the class of 4-desmethylsterols.

Plant stanols belong to the group of 4-desmethylsterols. Plant stanols are hydrogenation products of the respective plant sterols, e.g. campestanol/campesterol and sitostanol/sitosterol, and are found in nature at very low levels.

When edible oils undergo normal refining, plant sterols are partially extracted together with some tocopherols (in the process of natural vitamin E production). It is estimated that 2500 tonnes of vegetable oil needs to be refined to produce 1 tonne of plant sterols. Plant stanols are obtained by hydrogenation of the plant sterols.

Another source of plant sterols is tall oil, derived from the process of paper production from wood and approximately 2500 tonnes of pine is required to produce 1 tonne of plant sterols. Tall oil also contains a higher proportion of plant stanols (primarily β-sitostanol) than do vegetable oils.

Average dietary intakes of plant sterols in ’Western’ diets are between 200 - 400mg/day with vegetarians having higher intakes than those on mixed diets. Cooking oils, margarines and peanut butter (containing between 100 and 500mg/100g) have until recently been the main sources of plant sterols in typical Western diets. Legumes (up to 220mg/100g) and some seeds (e.g. sunflower and sesame 500 - 700mg/100g) are also good sources, while other vegetables and fruits contain slightly lower amounts of plant sterols. The intake of plant stanols is normally much lower. In nature, plant sterols can be in the free form or predominantly esterified with long chain fatty acids or with phenolic acids as in rice bran oil (ferulates) and shea butter (cinnamates). In the intestine, most sterol esters are hydrolysed to free sterols as part of the normal digestive process.

Evidence for plasma cholesterol lowering by plant sterol esters

It has been known for fifty years that plant sterols can lower blood cholesterol. Plant sterols have been the subject of numerous high dose (up to 25g/day), long term clinical trials to assess their effects on blood cholesterol levels. Over 1800 people in total have participated in these studies, conducted since the early 1950s (Pollak and Kritchevsky, 1981). From the 1950s to the 1980s, a preparation of predominantly β -sitosterol [Cytellin, Eli Lilly] was marketed in the USA to treat hypercholesterolaemia. No adverse effects were reported.

Yellow fat spreads and soft cheese products, supplemented with plant sterols and plant stanols, have appeared on the market. The consumption of average amounts of spread per day (approximately 20g in Western Europe) supplemented with between 8 - 10% plant sterol lowers serum total cholesterol and LDL cholesterol by 8 - 13% (Weststrate and Meijer,1998; Hendriks et al., 1999.) This is equivalent to the consumption of between 1.6 - 2.0g plant sterols per day. In an efficacy study on plant stanols a daily consumption of 24g of spread, containing 2-3g of plant stanol esters, total serum cholesterol and LDL cholesterol were lowered by up to 6.4% and 10.1% respectively (Nguyen et al., 1999). A dose dependant decrease in total and LDL cholesterol was observed with increasing levels of plant stanols esters up to a level of 1.6g/day. However, increasing the dose from 2.4g to 3.2g did not provide any clinically important additional effect (Hallikainen et al., 2000). Plant sterols and plant stanols appear to be equally effective in lowering plasma total and LDL cholesterol (Weststrate and Meijer, 1998; Law, 2000) and in a study with ileostomy subjects cholesterol absorption was found to be inhibited equally by esterified plant sterol and β-sitostanol esters (Normén et al., 2000).

In a clinical trial to examine the effects of plant sterols and plant stanols-enriched margarine on plasma lipid and phytosterol concentrations, de novo cholesterol synthesis and absorption rates in the context of a controlled diet, fifteen hypercholesterolemic men were fed either a diet of prepared foods alone or a diet containing 1.84 g plant sterols/stanols/day for 21 days in a cross-over study design. Dietary effects on total and LDL cholesterol concentrations were observed, although concentrations were lower with the plant sterol/stanol-enriched margarine than with the control diet on day 21 (P < 0.05). LDL-cholesterol concentrations on day 21 had decreased by 3.9% (P < 0.01) and 12.9% (P< 0.001) with the control diet and plant sterol-enriched margarine, respectively. For the plant stanol-enriched margarine, the decrease was 7.9% (p<0.01). High density lipoprotein (HDL)-cholesterol concentration did not change significantly (Jones et al., 2000).

Although people consume plant sterols every day in their normal diet, the amount is not great enough to have a significant blood cholesterol lowering effect. In order to achieve a cholesterol-lowering benefit, approximately 1 g/day of plant sterols need to be consumed (Hendriks et al., 1999).

Mechanisms for blood cholesterol lowering

The consumption of plant sterols and plant stanols lowers blood cholesterol levels by inhibiting the absorption of dietary and endogenously-produced cholesterol from the small intestine and the plant sterols/stanols are only very poorly absorbed themselves. This inhibition is related to the similarity in physico-chemical properties of plant sterols and stanols and cholesterol and it is generally accepted that there are two possible mechanisms by which this inhibition occurs:

The co-precipitation of cholesterol and plant sterols/stanols

In the intestinal lumen, cholesterol is found in solution with other fats. However, as monoglycerides and fatty acids are absorbed from the gut, concentration of the less well absorbed substances, e.g. sterols increases. When their concentration reaches a critical level, similar substances can precipitate from the solution. This may happen with cholesterol and plant sterols/stanols, because of their similarity in structure. Both cholesterol and plant sterols/stanols in the free state are poorly soluble in fat and micelles and in fact mutually limit each others solubility. Hence the greater the amount of plant sterols/stanols, the lower the solubility and perhaps the greater the amount of cholesterol precipitated. Cholesterol in the crystalline form cannot be absorbed.

The competition for space in mixed micelles

Micelle or mixed micelles are very efficient detergent-structures that solubilise the lipids secreted in the small intestine. Mixed micelles are composed of bile salts, phospholipids, tri, di- and monoglycerides, fatty acids, free cholesterol and fat soluble micronutrients. As there is only limited capacity in the micelles for carrying cholesterol, compounds with similar structures to cholesterol such as plant sterols and stanols, can compete with cholesterol for this space. Therefore increasing the amounts of plant sterols and stanols results in less and less cholesterol in mixed micelles and thereby decreased absorption of cholesterol from the gut.

In the process of absorption, cholesterol in the micelles is transported from the lumen of the small intestine across the intestinal mucous and into the lymph. The precise mechanism of cholesterol transport from the micelles into the intestinal cells is not fully understood. However, it is known that micelles are not absorbed intact, but that the fats, including cholesterol, pass through the brush border membrane into the cells, probably with the involvement of binding proteins and passive transport mechanisms.

Although people consume plant sterols every day in their normal diet, the amount is not great enough to have a significant blood cholesterol lowering effect. In order to achieve a cholesterol-lowering benefit, approximately 1 g/day of plant sterols need to be consumed (Hendriks et al., 1999).

Phytosterols protective against cancer?

Awad et al (2000a) reported on mice with human cancer tumours fed either a phytosterol diet or a cholesterol diet. Tumour size in animals fed the phytosterols was 33% smaller and had 20% fewer shifts of cancer cells to lymph nodes and lungs than in the cholesterol diet group.

Several theories regarding the mechanism of action of phytosterols as a protective factor include inhibiting of cell division, stimulating death of tumour cells and modifying some of the hormones that are essential to tumour growth [Awad et al (2000b), (2000c) (2003); Li et al (2001)].

Safety considerations

The blood cholesterol-lowering effect of plant sterols has been investigated in a large number of clinical trials on over 1800 people, with up to 25g per day and durations up to three years. No significant adverse effects have been observed in the decades of medically supervised clinical efficacy testing and the general clinical use of plant sterols.

In addition, both plant sterols and plant stanols have been subjected to a thorough toxicological evaluation. Studies on the absorption, distribution, metabolism and excretion have shown that plant sterols are poorly absorbed from the intestine (1 - 10%,) and that portion which is absorbed is rapidly eliminated via excretion in the bile (Sanders et al., 2000).

In a 13 week feeding study of plant sterol esters in rats (Hepburn et al., 1999), no adverse effects were seen at levels up to 3900mg/kg body weight/day (8.1% plant sterol esters in the diet; equivalent to 5.0% free plant sterol), the highest level tested. In a similar study plant stanol esters were reported to have no adverse effects at levels up to 1.78%, equivalent to 1.0% free stanols (Turnbull et al., 1999).

There are some suggestions from the literature that plant sterols are oestrogenic (Mellanen et al., 1996). However, in a series of in vitro assays with vegetable oil plant sterols it has been shown that they do not bind to either the human oestrogen receptor or the immature rat oestrogen receptor. Furthermore, there was no indication of oestrogenicity in an in vivo rat uterotrophic assay (Baker et al., 1999). Plant stanol esters have similarly been proven not to possess any oestrogenic potential in an in vitro assay (proliferation of MCF-7 cells) or in an in vivo rat uterotrophic assay (Turnbull et al., 1999). The reproductive toxicity of both plant sterols and plant stanols has also been assessed in a two generation reproductive toxicity study in the rat. There were no adverse effects of plant sterols up to approximately 4000mg/kg/day (8.1% vegetable oil sterol esters in the diet; equivalent to 5% free plant sterols); highest level tested (Waalkens-Berendsen et al., 1999). No adverse findings were reported for plant stanol esters up to levels of 4.38% in the diet, equivalent to 2.5% free stanols (Whittaker et al., 1999).

A series of human studies with vegetable oil plant sterol esters in spreads with intakes of up to 8.6g of plant sterols/day for 4 weeks have been conducted. Clinical chemistry, haematology, bacterial profile of the gut microflora and general physical condition were evaluated and no adverse effects were detected (Weststrate and Meijer, 1998; Weststrate et al., 1999s; Hendriks et al., 1999; Ayesh et al., 1999).

There is very rare condition, known as phytosterolaemia (sitosterolaemia), in which suffers have an inborn error in the metabolism of phytosterols. Only a few dozen cases are known worldwide and, for example, it has been reported that in The Netherlands there is only one known case. Due to the nature of the disease, sufferers are aware of their condition and should avoid additional intake of phytosterols.

Legislation

In the United States a panel of independent experts has concluded that vegetable oil sterol esters meeting appropriate food-grade specifications and produced by current good manufacturing practice (21 C.F.R. § 182.1(b)), are safe for use as an ingredient in vegetable oil spreads in amounts not to exceed 20%. It is the Panel’s opinion that qualified experts in the field would generally recognise that vegetable oil sterol esters are safe for this use, i.e. that vegetable oil sterol esters are generally recognised as safe (GRAS). The US Food and Drug Administration (FDA) have also cleared a spread containing up to 20% of plant sterol ester, and one containing plant stanol ester, on the basis of the GRAS recognition.

In Switzerland authorisation for Becel pro.activ, a special spread enriched with plant sterols, was given in September 1999 by the Swiss Health Office (Bundesamt fur Gesundheit; BAG).

In the European Union (EU) plant sterol esters for use in margarines/spreads has been reviewed under the EU Novel Foods Regulation (Regulation (EC) No 25 8/97) and the EU Scientific Committee on Food (SCF) concluded that the use of phytosterol-esters in yellow fat spreads (maximum level of 8% free phytosterols) is safe for human use. In addition, the SCF has expressed a general view on the long-term effects of the intake of elevated levels of phytosterols from multiple dietary sources. The Committee concluded that a numerical upper level for the total daily intake of phytosterols could not be established on the basis of the data available. In consideration of the dosages found to be effective for cholesterol lowering, without evidence of additional benefit at higher intakes and the possibility that high intakes might induce undesirable effects, it was considered prudent to avoid plant sterol intakes exceeding 3 g/day (SCF, 2002a). Furthermore, in the evaluation of the first application concerning plant sterols in yellow fat spreads, the SCF required that the applicant should establish a surveillance program accompanying the marketing of the product to obtain data on consumption and for further investigation of possible health effects, including among others the effects on plasma β-carotene levels. The results of this task have also been assessed by the SCF in its opinion on a report on post-launch monitoring of yellow fat spreads with added phytosterol esters (SCF, 2002b).

However, a yellow fat spread containing plant stanol esters was already legally on the market in the European Union without being subjected to review, because it was marketed in a Member State before the Novel Foods Regulation came into force. A similarly-based cheese spread by the same manufacturer considering it as merely a variant of the yellow fat spread, is also on the market in the UK, but was withdrawn from the shops in The Netherlands on instructions from the Dutch authority which regarded it as a separate novel food requiring prior approval.

The UK Advisory Committee on Novel Foods and Processes (ACNFP), responsible for assessing the safety of these products, asked the then Food Advisory Committee (FAC) to advise on the associated labelling issues. FAC concluded that the products should be clearly labelled so that those with an inborn error in the metabolism of phytosterols (phytosterolaemia) could avoid them. It also agreed that consumers should be informed that phytosterol ester-containing products are not nutritionally appropriate for young children and breast feeding women (as they did not need to reduce their blood cholesterol levels and there was a possibility that the products could affect vitamin A status). It stressed that this did not present a safety hazard and that the advice was purely for nutritional benefit rather than a health warning. Initially it recommended that the most appropriate way to do this would be through information circulated via GPs and other health professionals and through suitable magazines. Subsequently, however, it modified its earlier view and recommended that this information should also be provided on the product labels. This advice was welcomed by the then Committee on Medical Aspects of Food and Nutrition Policy (COMA).

More recently the Scientific Panel on Dietetic Products, Nutrition and Allergies of the European Food Safety Authority (EFSA) has recommended that sterol-containing foodstuffs should not be consumed in amounts resulting in total phytosterol intakes exceeding 3 g/day. With regard to β-sitostanol and campestanol, higher contents of 35% and 15%, respectively, were accepted by the Panel since these compounds or their esters have been sufficiently tested toxicologically. The Panel also reiterated the recommendations expressed in the previous SCF opinions in relation to the need for risk management measures to minimise the likelihood of a daily intake exceeding 3 g phytosterols/phytostanols, the provision of appropriate information to consumers regarding the need for regular consumption of fruits and vegetables to address the potential β-carotene lowering effect of the product, and the particular circumstances of phytosterolaemic patients, people under cholesterol-lowering medication and of women during pregnancy and lactation.

Labelling

In December 2004 the EU Commission published Regulation 608/2004/EC concerning the labelling of foods and food ingredients with added phytosterols, phytosterol esters, phytostanols and/or phytostanol esters, requiring such products to be labelled with additional information including the words “with added plant sterols/plant stanols”. In England, the Food with Added Phytosterols or Phytostanols (Labelling) (England) Regulations 2004 were published on 15 December 2004 to provide for the enforcement in England of Regulation 608/2004/EC

References

Awad A B et al (2000a). Peanuts as a Source of β-sitosterol, a Sterol with Anticancer Properties. Nutrition and Cancer. 36(2),238-41.

Awad A B et al (2000b). Dietary Phytosterol Inhibits the Growth and Metastisis of MDA-MB-231 Human Breast Cancer Cells Grown in SCID Mice. Anticancer Research. 20,821-24.

Awad A B et al (2000c). Phytosterols as Anticancer Dietary Components: Evidence and Mechanism of Action. Journal of Nutrition, 130,2127-30.

Awad A B et al (2003). Effect of phytosterols on cholesterol metabolism and MAP kinase in MDA-MB-231 human breast cancer cells. J. Nutr. Biochem. 14,111-119.

Ayesh R et al (1999). Safety evaluation of phytosterol-esters. Part 5: Faecal-short-chain fatty acid and microflora content, faecal bacterial enzyme activity and serum female sex hormones in healthy normolipidemic volunteers consuming a controlled diet either with or without a phytosterol-ester enriched margarine. Food and Chemical Toxicology, (1999) 37, (12), 1127-1138..

Baker V A et al(1999). Safety evaluation of phytosterol esters. Part 1: Assessment of oestrogenicity using a combination of in vivo and in vitro assays. Food and Chemical Toxicology, 37, 13-22.

EU Commission Regulation 608/2004/EC concerning the labelling of foods and food ingredients with added phytosterols, phytosterol esters, phytostanols and/or phytostanol esters

EU Scientific Committee on Food (2000). Opinion on a request for the safety assessment of the use of phytosterol esters in yellow fat spreads.

http://europa.eu.int/comm/food/fs/sc/scf/out56_en.pdf

EU Scientific Committee on Food (2002a). General view on the long-term effects of the intake of elevated levels of phytosterols from multiple dietary sources, with particular attention http://europa.eu.int/comm/food/fs/sc/scf/out143_en.pdf

EU Scientific Committee on Food (2002b). Opinion on a report on Post Launch Monitoring of “yellow fat spreads with added phytosterol esters”.

http://europa.eu.int/comm/food/fs/sc/scf/out144_en.pdf

EU Scientific Committee on Food (2003a). Opinion on applications for approval of a variety of plant sterol-enriched foods.

http://europa.eu.int/comm/food/fs/sc/scf/out174_en.pdf

EU Scientific Committee on Food) (2003b). Opinion on an application from ADM for

approval of plant sterol-enriched foods.

http://europa.eu.int/comm/food/fs/sc/scf/out192_en.pdf

EU Scientific Committee on Food (2003c). Opinion on an application from MultiBene for approval of plant sterol-enriched foods.

http://europa.eu.int/comm/food/fs/sc/scf/out191_en.pdf

European Food Safety Authority (2003). Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies. EFSA Journal, 15, 1-12 .

http://www.efsa.eu.int/science/nda/nda_opinions/216/opinion_nda_01_en1.pdf

Hallikainen M A et al (2000). Plant stanol esters affect serum cholesterol concentrations of hypercholesterolemic men and women in a dose dependent manner. Journal of Nutrition, 130, (4), 767-76.

Hendriks H F J et al (1999). Spreads enriched with three different levels of vegetable oil sterols and the degree of cholesterol lowering in normocholesterolaemic and mildly hypercholesterolaemic subjects. European Journal of Clinical Nutrition, 53, 319-327.

Hepburn P A et al(1999). Safety evaluation of phytosterol-esters. Part 2: Subchronic 90-day oral toxicity on phytosterol-esters — a novel functional food. Food and Chemical Toxicology, 37, (4), 521-532.

Jones P J H et al (2000). Modulation of plasma lipid levels and cholesterol kinetics by phytosterol versus phytostanol esters. Journal of Lipid Research, 41, 697-705.

Law M (2000). Plant sterol and stanol margarines and health. British Medical Journal, 320, 861-864.

Li J.H et al (2001) Measurement variability of plasma ß-sitosterol and campesterol, two new biomarkers for cancer prevention. Europ. J. Cancer Prev. 10, 245-249.

Mellanen P et al (1996). Wood-derived estrogens: Studies in vitro with breast cancer cell lines and in vivo in trout. Toxicology and Applied Pharmacology, 136, 381-388.

Normén L et al (2000). Soy sterol esters and ß-sitostanol ester as inhibitors of cholesterol absorption in human small bowel. American Journal of Clinical Nutrition, 71, 908-13.

Nguyen T T et al (1999). Cholesterol-lowering effect of stanol ester in a US population of mildly hypercholestrolemic men and women: a randomised controlled trial. Mayo Clinical Proceedings, 74 (12) 1198-1206.

Pollak 0 J (1953). Reduction of blood cholesterol in man. Circulation, 7, 702-706.

PolIak 0 J and Kritchevsky, D (1981). Sitosterol. Monographs on atherosclerosis. Basel: S. Karger.

Sanders D J et al (2000). The safety evaluation of phytosterol-esters Part 6. The comparative absorption and tissue distribution of phytosterols in the rat. Food and Chemical Toxicology, 38 (6) 485-491.

Turnbull D et al (1999). 13 Week oral toxicity study with stanol esters in rats. Regulatory Toxicology and Pharmacology, 29, 216-226.

Turnbull D et al (1999). Short term tests of estrogenic potential of plant stanols and plant stanol esters. Regulatory Toxicology and Pharmacology, 29, 211-215.

UK Food with Added Phytosterols or Phytostanols (Labelling) (England) Regulations 2004.

Waalkens-Berendsen D H et al(1999). Safety evaluation of phytosterol-esters. Part 3: Two-generation reproduction study in rats with phytosterol esters — a novel functional food. Food and Chemical Toxicology, 37, 683-696. .

Weststrate J A and Meijer G W (1998). Plant sterol-enriched margarines and reduction of plasma total- and LDL-cholesterol concentrations in normocholesterolaemic and mildly hypercholesterolaemic subjects. European Journal of Clinical Nutrition, 52, 334-343.

Weststrate J A et al (1999). Safety evaluation of phytosterol esters. Part 4: Faecal concentrations of bile acids and neutral sterols in healthy normolipidaemic volunteers consuming a controlled diet either with or without a phytosterol ester-enriched margarine. Food and Chemical Toxicology, 37, 1063-1071.

Whittaker M H et al (1999). Two generation reproductive toxicity study of plant stanol esters in rats. . Regulatory Toxicology and Pharmacology, 29, 196-204.

Ralph Blanchfield | alfa
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