Risk assessment of local dermal effects and skin sensitisation under the EU Chemicals Regulation REACH: a proposal for a qualitative, exposure scenario specific, approach.

Within the framework of REACH, an assessment regarding local dermal effects and skin sensitisation should be performed for substances. Quantitative hazard information for these effects is often not available. Furthermore, it is difficult to relate the way in which animals are exposed in dermal toxicity studies directly to dermal exposure in practice. In the absence of quantitative information, a qualitative assessment for dermal effects is the most reasonable option. The qualitative approach as proposed in the REACH guidance recommends only general risk management measures (RMM) for three categories with a low, moderate and high identified hazard, without specifying which RMM are needed for a specific exposure scenario. We propose to differentiate frequency of exposure based on differences in activities and to compare measured and estimated local skin exposure levels with rules of thumb for evaluation of control of risks per hazard category. For workers, specific RMM regimes are assigned to each combination of hazard category and process category (PROC). For consumers, a strategy in which RMM are arranged from product-integrated measures to the use of personal protective equipment (PPE) is presented. Our approach may be transferred into automated assessment tools like Chesar and CEFIC GES.

REACH, non-testing approaches and the urgent need for a change in mind set.

The objectives of REACH cannot be achieved under the current risk assessment approach. A change in mind set among all the relevant stakeholders is needed: risk assessment should move away from a labor-intensive and animal-consuming approach to intelligent and pragmatic testing, by combining exposure and hazard data effectively and trying to group chemicals (category approaches). The focus should be on reducing the overall uncertainties of 30,000 chemicals while acknowledging the existence of the uncertainty paradox: reducing uncertainty in the assessment of individual chemicals following the classical chemical-by-chemical approach as we have in previous decades will result in a prolongation of uncertainty for the entire group of 30,000 chemicals as a whole. With the first REACH registration deadline (2010) rapidly approaching, a mind set change is urgently needed. We can speed up the regulatory acceptance process, starting with the maximum use of currently available exposure and hazard data, tools and models. Optimal use should also be made of experimental exposure and hazard data generated under REACH. Only such an approach will make it possible to obtain a sufficient level of information within the time frame of REACH. A much more intensive dialogue between stakeholders is necessary.

Effect of moderate alcohol consumption on adipokines and insulin sensitivity in lean and overweight men: a diet intervention study.

OBJECTIVE: Moderate alcohol consumption is associated with a decreased risk of type II diabetes. This study investigates the effect of moderate alcohol consumption on adipokines and insulin sensitivity. SUBJECTS: Twenty healthy, lean (body mass index (BMI) 18.5-25 kg/m(2); n=11) or overweight (BMI>27 kg/m(2); n=9) men (18-25 years). METHODS: Three cans of beer (40 g alcohol) or alcohol-free beer daily during 3 weeks. RESULTS: Adiponectin and ghrelin concentrations increased (P<0.01) by 11 and 8%, while acylation-stimulating protein (ASP) concentrations decreased by 12% (P=0.04) after moderate alcohol consumption. Concentrations of leptin and resistin remained unchanged. Insulin sensitivity by an oral glucose tolerance test (OGTT) was not affected by moderate alcohol consumption, but 2 h glucose concentrations were lower (P=0.01) after beer (4.5+/-0.1 mmol/l) than alcohol-free beer (4.9+/-0.1 mmol/l). Both free fatty acids and glucagon concentrations showed a stronger increase (P<0.01) after 90 min during OGTT after beer than alcohol-free beer. Changes of adiponectin were positively correlated (r=0.69, P<0.001), and changes of leptin (r=-0.53, P=0.016) and ASP (r=-0.43, P=0.067) were negatively correlated with changes of insulin sensitivity index. All these results did not differ between lean and overweight men. CONCLUSIONS: Moderate alcohol consumption increased adiponectin and ghrelin, while it decreased ASP concentrations both in lean and overweight men. These changes are in line with the hypothesized improvement of insulin sensitivity, but did not affect insulin sensitivity within 3 weeks of moderate alcohol consumption.

Lactulose stimulates calcium absorption in postmenopausal women.

Animal studies have indicated that calcium absorption is increased by lactulose, a synthetic disaccharide. Therefore, the influence of lactulose on calcium absorption was measured in postmenopausal women who may benefit from the possible enhancing effect of lactulose on calcium absorption. Twelve postmenopausal women drank 100 ml of water containing 5 or 10 g of lactulose or a reference substance at breakfast for 9 days. The three treatments were given according to a randomized, double-blind, cross-over design, separated by two 19-day wash-out periods. On the 8th day of each treatment period, 44Ca dissolved in orange juice was drunk immediately after the solution with the study substance and just before a standard breakfast with 162 mg of carrier calcium. Within half an hour, 48Ca was given intravenously. Based on isotope ratios measured in urine collected before and until 36 h after isotope administration, true fractional calcium absorption was calculated. Calcium absorption during the treatments with the reference substance, 5 g and 10 g of lactulose was (mean +/- SD) 27.7 +/- 7.7, 30.0 +/- 7.6, and 32.2 +/- 7.0, respectively. A significant difference in calcium absorption was found between the highest dose of lactulose and the reference treatment (p < 0.01). A significant linear trend was found between the dose of lactulose and its positive effect on calcium absorption. In conclusion, in postmenopausal women a 9-day consumption of lactulose increases calcium absorption in a dose-response way. More research is warranted to explore how lactulose stimulates calcium absorption and whether it is able to improve calcium balance and/or to attenuate the rate of aging bone loss.

Reduced bone mass in Dutch adolescents fed a macrobiotic diet in early life.

This study investigated the effect of a macrobiotic (vegan-type) diet, low in calcium and vitamin D, consumed in early life, on bone mineral during adolescence. Bone mineral content (BMC) and bone area were measured in 195 adolescents (103 girls, 92 boys) aged 9-15 years, using dual-energy X-ray absorptiometry. Ninety-three adolescents (43 girls, 50 boys) had followed a macrobiotic diet in childhood, and 102 (60 girls, 42 boys) were control subjects. After adjustment for bone area, weight, height, percent body lean, age, and puberty, BMC was significantly lower in macrobiotic subjects, in boys and girls, respectively, at the whole body, -3.4% and -2.5%, spine, -8.5% and -5.0%, femoral neck, -8.0% and -8.2%, midshaft radius, -6.8% and -5.6%, and also in girls, at the trochanter, -5.8% (p < 0.05). No group differences were observed at the wrist. Group differences were not explained by current calcium adjusted bone mass at age 9-15 years, observations which may hold important implications for fracture risk in later life.

Dietary calcium and bone density in adolescent girls and young women in Europe.

The objective of this study was to investigate the association between dietary calcium intake and radial bone density among young women, over the whole range of intake and at different levels of calcium intake. The study design was a cross-sectional, observational multicenter investigation in six European countries. One thousand one hundred and sixteen healthy Caucasian girls aged 11-15 years and 526 women aged 20-23 years participated, after having been selected from larger population samples to represent a large range in calcium intake. Bone mineral density (BMD) was measured with dual-energy X-ray absorptiometry at the ultradistal and middistal radius. Calcium intake was assessed with 3-day food records. Other potential determinants of BMD were measured by anthropometry or questionnaires. Mean calcium intake among the girls varied between 609 mg/day in Italy and 1267 mg/day in Finland; intakes for women were in a similar range. After adjustment for height, weight, and age at menarche for the women, and adjustment for age, height, weight, Tanner stage, and bone area for the girls, radial BMD at both sites did not significantly vary among quartiles of calcium intakes for both age groups. In multivariate linear regression, calcium was weakly positively associated with BMD at both sites in the girls (per 100 mg of calcium: beta = 0.57 mg/cm2, p = 0.03 for middistal BMD and beta = 0.56 mg/cm2, p = 0.01 for ultradistal BMD). For middistal BMD, the association was observed predominantly in pre-menarcheal girls. The associations were no longer statistically significant after full adjustment for all determinants of BMD, except again in pre-menarcheal girls. Radial BMD in the women was not associated with calcium intake, except after full adjustment for determinants of BMD, when ultradistal BMD became inversely associated with calcium intake (per 100 mg beta = -1.02, p = 0.03); this finding was due to results in one of the countries and not found in other countries. There was no evidence for a different relation between calcium and BMD at different levels of intake; although there was a positive association at calcium intake levels < 600 mg/day, the interaction was not significant and there was no consistent trend over intake categories. These results do not support the hypothesis that dietary calcium is a determinant of peak BMD in European women, for a wide range of intake. This study does not provide evidence that Recommended Dietary Allowances for calcium should be increased.

Effects of sugar intake on body weight: a review.

Weight reduction programmes are mainly focused on reducing intake of fat and sugar. In this review we have evaluated whether the replacement of dietary (added) sugar by low-energy sweeteners or complex carbohydrates contributes to weight reduction. In two experimental studies, no short-term differences in weight loss were observed after use of aspartame as compared to sugar in obese subjects following a controlled energy-restricted diet. However, consumption of aspartame was associated with improved weight maintenance after a year. In two short-term studies in which energy intake was not restricted, substitution of sucrose by artificial sweeteners, investigated mostly in beverages, resulted in lower energy intake and lower body weight. Similarly, two short-term studies, comparing the effect of sucrose and starch on weight loss in obese subjects did not find differences when the total energy intake was equal and reduced. An ad libitum diet with complex carbohydrates resulted in lower energy intake compared to high-sugar diets. In two out of three studies, this was reflected in lower body weight in subjects consuming the complex carbohydrate diet. In conclusion, a limited number of relatively short-term studies suggest that replacing (added) sugar by low-energy sweeteners or by complex carbohydrates in an ad libitum diet might result in lower energy intake and reduced body weight. In the long term, this might be beneficial for weight maintenance. However, the number of studies is small and overall conclusions, in particular for the long term, cannot be drawn.

Oral calcium and blood pressure: a controlled intervention trial.

In a double-blind, placebo-controlled trial with 58 normotensive female students, the effect of oral-calcium supplementation (1500 mg Ca++/day for 6 wk) on diastolic and systolic blood pressure was studied while students were consuming a low-calcium diet (500 mg Ca++/day) by restricting the intake of dairy products. Results show that, in both the calcium- and placebo-supplemented groups, blood pressure values decreased slightly and no effect of oral-calcium supplementation on blood pressure could be demonstrated. In addition, at baseline neither systolic nor diastolic blood pressure correlated with habitual calcium intake. Diastolic but not systolic blood pressure correlated significantly with body mass index (r = 0.31, p = 0.01). It is concluded that oral-calcium supplementation for 6 wk does not influence blood pressure in young, healthy normotensive females consuming low-calcium diet.

Nondigestible oligosaccharides do not interfere with calcium and nonheme-iron absorption in young, healthy men.

The effect of inulin, fructooligosaccharides, and galactooligosaccharides on true intestinal absorption of iron and calcium was measured in men by using double stable-isotope techniques (oral 57Fe and 44Ca and intravenous 58Fe and 48Ca). The incorporation of iron isotopes into erythrocytes and the urinary excretion of calcium isotopes was measured with an inductively coupled plasma mass spectrometer. Twelve healthy, nonanemic, male subjects aged 20-30 y received four treatments consisting of a constant basal diet supplemented with 15 g/d inulin, fructooligosaccharide, or galactooligosaccharide, or not supplemented (control treatment). These four treatments were given for 21 d each according to a randomized, crossover design. Iron absorption was measured over the last 7 d of treatment (days 15-21) and calcium absorption was measured on day 21 of each treatment period. Mean (+/- SEM) iron absorption was 5.5 +/- 1.6%, 6.1 +/- 1.9%, 5.3 +/- 1.9%, and 5.1 +/- 1.5%, respectively, during treatment with inulin, fructooligosaccharide, galactooligosaccharide, or the control; mean ( SEM) calcium absorption was 25.8 +/- 2.3%, 26.3 +/- 1.9%, 26.3 +/- 2.6%, and 28.1 +/- 4.3%, respectively. None of the differences between treatments was significant. It is concluded that 15 g/d inulin, fructooligosaccharide, or galactooligosaccharide did not have a negative effect on iron and calcium absorption in young healthy men.

Effect of protein source and quantity on protein metabolism in elderly women.

To study sequentially the effect of meal feeding and the effect of protein source and quantity on whole-body protein metabolism, 12 elderly women consumed 3 diets differing in both the quantity and source of protein (diet A: 5.3% of energy intake provided by animal protein and 5.0% by vegetable protein; diet B: 14.5% of energy provided by animal protein and 5.1% by vegetable protein; diet C: 5.0% of energy provided by animal protein and 15.1% by vegetable protein). The diets were consumed for 2 wk with a 2-wk interval between diets. At the end of each dietary period, nitrogen balance and protein turnover were measured. Protein turnover was measured during 4 h of fasting followed by 4 h of feeding. Comparisons were made between fasted and fed periods (within one diet) and between the diets to study the effect of the protein source and quantity. Mean nitrogen balance did not differ significantly from zero during diets B and C and was not affected by the protein source. Meal feeding resulted in increased protein flux and protein oxidation and decreased protein breakdown compared with the postabsorptive values; there was no effect of feeding on protein synthesis. With the high-vegetable-protein diet, protein breakdown in the absorptive state was not inhibited to the same extent as during the high-animal-protein diet, resulting in less net protein synthesis during the high-vegetable-protein diet than during the high-animal-protein diet.

Effects of prebiotics on mineral metabolism.

Nondigestible oligosaccharides (NDOs) have been found to stimulate absorption of several minerals and to improve mineralization of bone. Hence, these substances are potential ingredients for "functional foods." In addition to a nutritional effect, functional foods have physiologic and psychological benefits that result in improved health or reduced risk of chronic disease. Most of the scientific evidence for the functional effects of NDOs is based on animal experiments in which NDOs increased the availability of calcium, magnesium, zinc, and iron. This stimulatory effect of some NDOs is assumed to be mainly due to their prebiotic character. A prebiotic is defined as a substrate or food ingredient that is nondigestible for the host but is fermented selectively by some of the intestinal microflora. Thus, it stimulates the growth and activity of bacteria with beneficial consequences for the host's health. Recently, these findings were confirmed in human studies for some NDOs. The effects seem to be specific for the type of carbohydrate and are likely related to the rate of fermentation by the intestinal flora and appear to depend on the ingested dose. Contradictory results of the effect of prebiotics in literature may be due to the experimental design because the effect of NDOs depends on the dose, the time of administration, the content of calcium in the diet, the part of the skeleton investigated, and the age of the subjects studied.

Oligofructose stimulates calcium absorption in adolescents.

BACKGROUND: In rats, nondigestible oligosaccharides stimulate calcium absorption. Recently, this effect was also found in human subjects. OBJECTIVE: The objective of the study was to investigate whether consumption of 15 g oligofructose/d stimulates calcium absorption in male adolescents. DESIGN: Twelve healthy, male adolescents aged 14-16 y received, for 9 d, 15 g oligofructose or sucrose (control treatment) daily over 3 main meals. The treatments were given according to a randomized, double-blind, crossover design, separated by a 19-d washout period. On the 8th day of each treatment period, 44Ca was given orally with a standard breakfast containing approximately 200 mg Ca. Within half an hour after administration of 44Ca, 48Ca was administered intravenously. Fractional calcium absorption was computed from the enrichment of 44Ca:43Ca and 48Ca:43Ca in 36-h urine samples, which was measured by inductively coupled plasma mass spectrometry. RESULTS: An increase in true fractional calcium absorption (%) was found after consumption of oligofructose (mean difference +/- SE of difference: 10.8+/-5.6; P < 0.05, one sided). The results are discussed in relation to the methods used. CONCLUSION: Fifteen grams of oligofructose per day stimulates fractional calcium absorption in male adolescents.

The effect of dietary sodium on urinary calcium and potassium excretion in normotensive men with different calcium intakes.

On the basis of their usual calcium intake, 12 healthy normotensive male students were divided into a low and a high calcium group. Both groups were provided with complete diets containing the same calculated quantities, expressed per MJ, of protein, fat, carbohydrate, sodium, potassium, phosphorus, and calcium. After an initial period of two weeks during which all participants received an additional 100 mmol sodium daily, 6 participants received an additional 22 mmol sodium/day (low sodium), the other 6 participants 178 mmol sodium/day (high sodium). After two weeks the sodium regimes were crossed over for a subsequent two-week period. When the second week of the low sodium period is compared with the second week of the high sodium period, the increase in the calcium/creatinine ratio, expressed on molar basis, was 0.059 for the low and 0.053 for the high calcium group (p less than 0.05). The increase in the molar potassium/creatinine ratio was 0.65 for the low and 0.03 for the high calcium group (p less than 0.025). However, after correcting for the calcium intake the effect in the low calcium group was found to be greater (p less than 0.05). Sodium supplementation was found to have no effect on blood pressure, but mean systolic (p less than 0.10) and diastolic in the high calcium group were lower. This suggests that calcium may well play a role in the regulation of blood pressure.

Design and data quality of a mixed longitudinal study to elucidate the role of dietary calcium and phosphorus on bone mineralization in pre-, peri-, and postmenopausal women.

The study design and data quality control of an ongoing study (10 yr duration) in a few hundred women are presented. Good variables with respect to their longitudinal usefulness are: body weight, body height, and span-width. Reasonable variables are the bone parameters of the radius (BMC, BW, and BMC/BW). Poor variables are: dietary calcium and phosphorus intake, dietary calcium-to-phosphorus ratio, urinary calcium-to-creatinine ratio, urinary sodium-to-creatinine ratio, hematocrit, serum alkaline phosphatase activity, serum gamma-GT activity, and serum parathyroid-hormone concentration. Bad variables are: urinary phosphorus-to-creatinine ratio, urinary hydroxyproline-to-creatinine ratio, creatinine clearance, hemoglobin, MCHC, serum calcium, serum ionized calcium, serum phosphorus, serum total protein, serum albumin, and serum creatinine. In conclusion, it is possible to relate bone loss to food intake and to changes in anthropometric variables on an individual basis. However, quantification of the metabolic process is not possible.

Effect of dietary calcium supplementation with lactose on bone in vitamin D-deficient rats.

The relationship between plasma calcium and bone length, chemical and histomorphometric bone parameters was studied in vitamin D-deficient rats in order to determine whether the effects of vitamin D on bone could be attributed to the effect of vitamin D on serum calcium. Plasma calcium was varied over a wide range of dietary manipulation. Four groups of vitamin D-deficient rats were given for 6 weeks: a vitamin D-deficient diet (D-, n = 6), the D--diet with calcium supplementation (D-Ca+, n = 6), the D-Ca+-diet with lactose substituted for dextrose (D-Ca+lac, n = 6) or a normal diet (D+, n = 8). After 6 weeks the mean plasma calcium concentrations were 6.1; 7.0; 9.8; and 10.4 mg/dl, respectively. In the vitamin D-deficient rats (groups D-, D-Ca+, D-Ca+lac) plasma calcium was correlated with bone length, bone ash, volumetric density of osteoid in the metaphysis of the tibia, and volumetric density of trabecular bone in the same bone section. In the D-Ca+lac group these bone parameters approximated the values of the D+ group, but were still significantly lower. It is concluded that in vitamin D-deficient rats longitudinal bone growth, bone mineral content and bone histomorphometry can be brought close to normal by supplying additional dietary calcium with lactose, without vitamin D repletion. The study does not exclude the possibility that residual amounts of vitamin D are required to obtain this effect.

The influence of dietary calcium on kidney calcification and renal function in rats fed high-phosphate diets.

This study was carried out to test the hypothesis that kidney calcification caused by high dietary phosphorus intake can be prevented by increasing the calcium content of the diet, because the latter reduces the intestinal P absorption. The investigation was conducted in rats fed semipurified diets with either a low (0.15%) or a high (1.20%) P content. Although dietary Ca supplementation (as CaCO3 or CaCl2) reduced the intestinal P absorption, it did not prevent kidney calcification induced by high dietary P. On the contrary, when CaCO3 was supplemented for 16 weeks, renal calcification increased, probably due to the alkaline nature of this salt. Both CaCO3 and CaCl2 in the high-P diets adversely influenced Mg use. This might explain the failure of these salts to prevent kidney calcification.

Daily moderate alcohol consumption increases serum paraoxonase activity; a diet-controlled, randomised intervention study in middle-aged men.

Moderate alcohol consumption is associated with a reduced risk of coronary heart disease. Part of this inverse association may be explained by its effects on HDL. Paraoxonase, an HDL-associated enzyme, has been suggested to protect against LDL oxidation. We examined the effects of moderate consumption of red wine, beer and spirits in comparison with mineral water on paraoxonase activity in serum. In this diet-controlled, randomised, cross-over study 11 healthy middle-aged men consumed each of the beverages with evening dinner for 3 weeks. At the end of each 3 week period, blood samples were collected pre- and postprandially and after an overnight fast. Fasting paraoxonase activity was higher after intake of wine (P<0. 001), beer (P<0.001), and spirits (P<0.001) than after water consumption (149.4+/-111.1, 152.6+/-113.1, 152.8+/-116.5 and 143. 1+/-107.9 U/l serum), but did not differ significantly between the 3 alcoholic beverages. Similar effects were observed pre- and postprandially. The increases in paraoxonase activity were strongly correlated with coincident increases in concentrations of HDL-C and apo A-I (r=0.60, P<0.05 and r=0.70, P<0.05). These data suggest that increased serum paraoxonase may be one of the biological mechanisms underlying the reduced coronary heart disease risk in moderate alcohol consumers

Effects of moderate alcohol consumption on platelet function, tissue-type plasminogen activator and plasminogen activator inhibitor.

Short-term effects of moderate alcohol consumption on platelet function, tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) were studied in two age groups of volunteers (20-30 and 45-55 years), each consisting of eight healthy males. The alcohol (30 g in red port and wine) was consumed during a standard dinner. Two blood samples were drawn: one in the postprandial phase, and one the next morning after fasting overnight. Alcohol consumption tended to increase platelet aggregation and production of hydroxy fatty acids, reduced plasma t-PA activity and increased PAI activity in the postprandial phase. After the overnight fast the effects on t-PA and PAI had disappeared whereas at that time alcohol consumption tended to decrease platelet function. The effects of alcohol on t-PA and PAI activity appeared mainly in the older age group, whereas the t-PA activity in this group was already much lower, irrespective of alcohol consumption.

The effect of consumption of milk fermented by Lactobacillus casei strain Shirota on the intestinal microflora and immune parameters in humans.

OBJECTIVE: To determine the effect of consumption of milk fermented by Lactobacillus casei strain Shirota (L. casei Shirota) on the composition and metabolic activities of the intestinal microflora, and immune parameters in humans. SUBJECTS: Twenty healthy male subjects aged 40-65 years were selected. DESIGN: A placebo-controlled trial was performed in which 10 subjects were randomly assigned to a control and 10 to a treatment group. During the first and last two weeks of the 8-week study the subjects received a strictly controlled diet without fermented products. The same controlled diet was given during the intermediate 4-week test period but then the treatment group received three times daily 100 ml of fermented milk containing 10(9) CFU L. casei Shirota/ml, whereas the same amount of unfermented milk was given to the subjects in the control group. RESULTS: In comparison to the control group, the consumption of L. casei Shirota-fermented milk resulted in an increase of the Lactobacillus count in the faeces in which the administered L. casei Shirota was predominant at the level of 10(7) CFU/g wet faeces. This was associated with a significant increase in Bifidobacterium counts (P < 0.05). Some shifts in the other bacterial species were found, such as a decreased number of Clostridium; however the differences were not statistically different between the treatment and the control groups. The beta-glucuronidase and beta-glucosidase activities per 10(10) bacteria decreased significantly (P < 0.05) at the second week of the 4-week test period with the consumption of L. casei Shirota-fermented milk. Furthermore, the consumption of the fermented milk product resulted in a slight but significant increase in the moisture content of the faecal samples (P < 0.05). No treatment effects were observed for any of the immune parameters measured (including natural killer (NK) cell activity, phagocytosis and cytokine production). CONCLUSIONS: The results suggest that consumption of L. casei Shirota-fermented milk is able to modulate the composition and metabolic activity of the intestinal flora and indicate that L. casei Shirota-fermented milk does not influence the immune system of healthy immunocompetent males.

Effects of a milk product, fermented by Lactobacillus acidophilus and with fructo-oligosaccharides added, on blood lipids in male volunteers.

OBJECTIVE: To investigate in adult male volunteers the effect of a new fermented milk product, fermented by Lactobacillus acidophilus and with fructo-oligosaccharides added, on blood lipids. DESIGN: Randomized placebo-controlled double-blind two-way cross over trial with two treatment periods of three weeks, separated by a wash-out period of one week. SETTING: the study was performed at the TNO Nutrition and Food Research Institute in Zeist, The Netherlands. SUBECTS: Thirty normal healthy men, aged 33-64y (mean serum total cholesterol level: 5.23 +/- 1.03 (s.d.)), were selected for this study. Normal health was assessed by pre study screening. All subjects were used to an average Dutch food pattern. INTERVENTIONS: During the treatment periods subjects consumed three times daily a 125 ml of either test or reference product as a part of their habitual diet. The test product was a milk, fermented by yogurt starters and Lactobacillus acidophilus, and contained 2.5% fructo-oligosaccharides, 0.5% vegetable oil and 0.5% milk fat. The reference product was a traditional yogurt (milk fermented only by yogurt strains), containing 1% milk fat. Blood samples for serum lipid analysis and blood glucose measurements were taken before the start of the experiment and at the end of both treatment periods. RESULTS: As compared to the reference product, consumption of the test product resulted in significantly lower values for serum total cholesterol (P < 0.001), LDL-cholesterol (P < 0.005), and the LDL/HDL-ratio (P < 0.05) by 4.4, 5.4 and 5.3% respectively. Levels of serum HDL-cholesterol, triglycerides and blood glucose remained essentially unchanged. The beneficial effects of the test product on serum cholesterol were largely related to an increase of this parameter during the consumption of the reference product. CONCLUSIONS: As compared to traditional yogurt, daily consumption of three times 125 ml of test product specifically lowered serum LDL-cholesterol levels in normal healthy male adult subjects with borderline elevated levels of serum total cholesterol within three weeks.