[Validation of the French version of the Binge Eating Scale: Examination of its factor structure, internal consistency and construct validity in a non-clinical and a clinical population]. / Validation de la version française de la Binge Eating Scale : étude de sa structure factorielle, de sa consistance interne et de sa validité de construit en population clinique et non clinique.

OBJECTIVE: The Binge Eating Scale is a widely used scale to assess binge eating disorder in obese patients. Until now, this scale has not been validated on a French population, and no psychometrically sound tool assesses binge eating disorder in the French. This study aimed to test the psychometric properties of a French version of the Binge Eating Scale by establishing its factor structure, internal consistency, and construct validity in both a non-clinical population and a clinical population (obese patients who are candidates for bariatric surgery). METHODS: A total of 553 non-clinical subjects and 63 morbidly obese patients who were candidates for bariatric surgery were assessed with the BES and the Bulimic Investigatory Test, Edinburgh or BITE (which assesses both binge eating behaviours and use of inappropriate compensatory behaviours). We tested the factor structure of the instrument, its internal consistency, its construct validity with measures of binge eating, and its construct validity with measures of inappropriate compensatory behaviours to avoid weight gain. In 47 out of the 63 obese patients, we assessed binge eating disorder (SCID). RESULTS: In the non-clinical population, the BES had a one-factor structure (which accounted for 61% of the variance), excellent internal consistency (α=0.93), and high construct validity with measures of binge eating. In this population, construct validity with measures of inappropriate compensatory behaviours was confirmed in overweight and obese subjects (P=0.42), but not in underweight and optimal weight subjects (P<0.001). In obese patients candidates for bariatric surgery, we demonstrated that the BES had a one-factor structure (which accounted for 46% of the variance), had high internal consistency (α=0.88) and high construct validity with measures of binge eating and good construct validity with measures of inappropriate compensatory behaviours to avoid weight gain. In the subpopulation of 47 obese patients, sensitivity, specificity, positive predictive value and negative predictive value were respectively 75%, 88.4%, 37.5% and 97.4% (BES threshold=18). DISCUSSION: In this study, we validated a psychometrically sound French version of the Binge Eating Scale, both in a non-clinical and a clinical sample. The psychometric properties of the French version of the BES are comparable to its original version with a one-factor structure. The BES is a useful tool to assess binge eating disorder in obese patients (e.g., bariatric surgery candidates), but might not differentiate between binge eating disorder and bulimia nervosa in underweight and optimal weight subjects.

[Diabetic amyotrophy: favorable outcome following corticosteroid therapy?]. / Amyotrophie diabétique : intérêt de la corticothérapie ?

Diabetic amyotrophy or lombosacral radiculoplexus neuropathy is a rare complication associated with early-stage diabetes. Thigh pain, quadricipital amyotrophy, proximal weakness of lower limbs and weight loss are the main symptoms of the disease. As neurological damage is related to inflammatory microvasculitis, corticosteroid therapy may be considered as the first line therapy. We report a 54-year-old patient with type 2 diabetes affected with severe diabetic amyotrophy. Following intravenous corticosteroid therapy, the patient reported a rapid pain relief and gained muscle strength.

Physical activity and weight loss following bariatric surgery.

Physical activity is a cornerstone in the medical management of obesity and could be important for weight loss following bariatric surgery. This review aims to describe the evolution of physical activity following massive weight loss induced by bariatric surgery, and to identify the relationship between physical activity and amount of weight loss. A literature search identified 20 publications (19 studies) reporting physical activity data in relation to bariatric surgery. All studies were observational. Self-assessment of physical activity was used in all the studies. Objective measures (pedometry) were used in two studies. The time frame for physical activity assessment varied: before surgery in two publications, after surgery in nine, and longitudinal pre- to post-operative evolution in nine. The latter nine publications found an increase in physical activity after bariatric surgery. In 10/13 studies where it was described, there was a positive relationship between physical activity level and amount of weight loss. In conclusion, observational evidence of self-reported physical activity suggests that physical activity increases after bariatric surgery and that physical activity is associated with surgically induced weight loss. However, these findings warrant further evaluation using objective measures of physical activity and testing in controlled trials.

The cumulative effect of small dietary changes may significantly improve nutritional intakes in free-living children and adults.

BACKGROUND/OBJECTIVES: The ELPAS (Etude Longitudinale Prospective Alimentation et Santé) study was an 8-month randomized controlled dietary modification trial designed to test the hypothesis that family dietary coaching would improve nutritional intakes and weight control in 2026 free-living children and parents. It resulted in significant nutritional changes, with beneficial effects on body mass index in adults. In these ancillary analyses, we investigated dietary changes throughout the intervention. SUBJECTS/METHODS: Before the study, modeling analyses were carried out on the French Association Sucre Produits Sucrés Consommation et Communication (ASPCC) food-consumption database to identify the most efficient dietary intervention strategy. During the study, all participants performed monthly three nonconsecutive 24-h dietary recalls: this allowed for measuring changes in the number of servings per day and serving size for each targeted food category throughout the intervention. RESULTS: Modeling analyses showed that targeting only the 10 main foods contributing to fat and carbohydrate intakes did not allow for reaching the ELPAS nutritional goals. As a result, it was decided to target more foods and to propose several types of dietary advice (such as change in serving size, change in cooking method, food substitution). This strategy led to many appropriate dietary changes during the intervention, but only a few of them reached significance. The mean number of servings per day was indeed significantly modified for only 7% of the targeted food categories in children and 17% in parents. The mean serving size was modified for only 12% of targeted food categories in children and 9% in parents. CONCLUSIONS: The cumulative effect of small dietary changes may induce significant nutritional improvements, with limited burden for populations.

Improving outcome of chemotherapy of metastatic breast cancer by docosahexaenoic acid: a phase II trial.

BACKGROUND: Breast cancer becomes lethal when visceral metastases develop. At this stage, anti-cancer treatments aim at relieving symptoms and delaying death without resulting in additional toxicity. On the basis of their differential anti-oxidant defence level, tumour cells can be made more sensitive to chemotherapy than non-tumour cells when membrane lipids are enriched with docosahexaenoic acid (DHA), a peroxidisable and oxidative-stress-inducing lipid of marine origin. METHODS: This open-label single-arm phase II study evaluated the safety and efficacy (response rate), as primary end points, of the addition of 1.8 g DHA daily to an anthracycline-based chemotherapy (FEC) regimen in breast cancer patients (n = 25) with rapidly progressing visceral metastases. The secondary end points were time to progression (TTP) and overall survival (OS). RESULTS: The objective response rate was 44%. With a mean follow-up time of 31 months (range 2-96 months), the median TTP was 6 months. Median OS was 22 months and reached 34 months in the sub-population of patients (n = 12) with the highest plasma DHA incorporation. The most common grade 3 or 4 toxicity was neutropaenia (80%). CONCLUSION: DHA during chemotherapy was devoid of adverse side effects and can improve the outcome of chemotherapy when highly incorporated. DHA has a potential to specifically chemosensitise tumours.

Changes in plasma triacylglycerol concentrations after sequential lunch and dinner in healthy subjects.

OBJECTIVES: The present study examines the kinetic of plasma triacylglycerol (TAG) after sequential ingestion of lunch and dinner as well as the contribution of dietary fat ingested at lunch to subsequent post-dinner TAG composition. METHOD: Six healthy subjects were included. After standardized breakfast (7: 30AM), 2 mixed meals with fat loads composed of 44 g olive oil (rich in oleic acid) at lunch (12PM) and 44 g sunflower oil (rich in linoleic acid) at dinner (7PM) were ingested. [1-13C] palmitate was added in lunch only. Plasma TAG and chylomicron-TAG (CMTAG) levels were measured sequentially after meals. [1-13C] palmitate enrichment and concentrations of oleic acid and linoleic acid were measured in all lipid fractions. RESULT: Post-dinner plasma TAG peak was delayed as compared to lunch (3 hours vs 1 hour, p=0.002) whereas the magnitude of the postprandial peaks was not significantly different between lunch and dinner (2.4+/-0.3 vs 2.0+/-0.4 mmol/L, p=0.85). [1-13C] palmitate enrichment was maximal 5 hours after lunch in all lipid fractions and decreased slowly thereafter. After dinner ingestion, the rate of decline of [1-13C] palmitate enrichment plateaued during the first 60 minutes. Oleic acid increased slightly and immediately after dinner and remained the predominant fatty acid in all lipid fractions during the first hour after dinner. A delayed peak of plasma and CM-TAG was observed after dinner as compared to lunch without difference in the magnitude of peaks. CONCLUSION: The contribution of dietary fat ingested at lunch to post-dinner lipemia is confirmed despite the relatively long lasting interval between the 2 meals (7 h) and the absence of any early peak of plasma TAG after dinner.

[Maternal mortality estimation in France, according to a new method]. / Estimation de la mortalité maternelle en France: une nouvelle méthode.

OBJECTIVE: Ten years after implementation of maternal mortality monitoring in France, we established a new estimate of the current maternal mortality ratio (MMR) and revisited maternal death data collection. MATERIAL AND METHODS: Linkages were set up between female deaths and childbirths and between female deaths and causes of death. Information provided by confidential inquiries into maternal deaths carried out by the National Committee for maternal mortality study was added. The World Health Organization (WHO) definitions were used for maternal death and maternal mortality ratio. The study concerned deaths occurring in 1999. Results were compared with data from 1989. RESULTS: The official data showed 20% fewer maternal deaths than our inquiry. Estimated from our data, the MMR was 9 per 100000 live births in 1999. Direct obstetric causes were more often recorded than indirect causes. Hemorrhage was the leading obstetric cause of maternal death (21%). In comparison with the 1989-90 data, the underestimation of maternal deaths and maternal mortality ratios are improving (from 18 to 9 per 100000). CONCLUSION: These results, obtained while the mean maternal age at childbirth increased regularly, are interpreted as a sign of improvement in care. But the persistence of post partum hemorrhages as the leading cause of maternal death and the high rate of avoidable deaths, disclose important targets for further progress.

"Big IGF-II"-induced hypoglycemia secondary to gastric adenocarcinoma.

Non-islet cell tumor-related hypoglycemia is a rare phenomenon. We report the case of a 63 Year-old man admitted for hemiparesia and a capillary blood glucose of 20 mg/dL. The presence of an immature form of IGF-II that can mimic the effect of insulin, namely "big IGF-II", explained this patient's hypoglycaemia. A moderately differentiated adenocarcinoma of the cardia with metastatic extension to the stomach and the liver was demonstrated. Octreotide failed to control the hypoglycaemia, therefore prednisolone (2 mg/kg per day) and enteral feeding prevented new episodes of severe hypoglycaemia.

Association between trans fatty acid intake and cardiovascular risk factors in Europe: the TRANSFAIR study.

BACKGROUND: High intakes of trans fatty acids (TFA) have been found to exert an undesirable effect on serum lipid profiles, and thus may increase the risk for cardiovascular disease. OBJECTIVES: Investigation of the association between TFA intake and serum lipids. DESIGN: Cross-sectional study in eight European countries (Finland, France, Greece, Iceland, The Netherlands, Portugal, Spain, Sweden) among 327 men and 299 women (50-65 y). Using a dietary history method, food consumption was assessed and TFA intake was calculated with recent figures on TFA levels of foods, collected in the TRANSFAIR study. RESULTS: Mean (+/-s.d.) TFA intake was 2.40+/-1.53 g/day for men and 1.98+/-1.49 g/day for women (0.87+/-0.48% and 0. 95+/-0.55% of energy, respectively), with the highest consumption in Iceland and the lowest in the Mediterranean countries. No associations were found between total TFA intake and LDL, HDL or LDL/HDL ratio after adjustment for cardiovascular risk factors. Additional adjustment for other fatty acid clusters resulted in a significant inverse trend between total TFA intake and total cholesterol (Ptrend<0.03). The most abundantly occurring TFA isomer, C18:1 t, contributed substantially to this inverse association. The TFA isomers C14:1 t9, C16:1 t9 and C22:1 t were not associated or were positively associated with LDL or total cholesterol. CONCLUSIONS: From this study we conclude that at the current European intake levels of trans fatty acids they are not associated with an unfavourable serum lipid profile. SPONSORSHIP: Unilever Research Laboratorium, the Dutch Dairy Foundation on Nutrition and Health, Cargill BV, the Institute of Food Research Norwich Laboratory, the Nutrition Branch of the Ministry of Agriculture, Fisheries and Food, the International Fishmeal and Oil Manufacturers' Association, Kraft Foods, NV Vandemoortele Coordination Center, Danone Group, McDonalds Deutschland Inc, Danish Veterinary and Food Administration, Valio Ltd, Raisio Group. European Journal of Clinical Nutrition (2000) 54, 126-135

Effects of an amino acid dialysate on leucine metabolism in continuous ambulatory peritoneal dialysis patients.

BACKGROUND: Protein-energy malnutrition is frequent in continuous ambulatory peritoneal dialysis (CAPD) patients. The use of amino acids in the dialysate could improve the protein balance, especially if associated to a concomitant energy intake. METHODS: A 1.1% amino acid solution for peritoneal dialysis was administered to CAPD patients over 30 minutes during concomitant absorption of 600 ml water (control study) or of a 600 kcal meal/600 ml. Leucine metabolism was studied using the combination of intravenous [2H3] and intraperitoneal [13C] leucine. RESULTS: The rate of leucine appearance was stimulated by 56 and 53% (control and meal) at 45 minutes. The rates of leucine appearance and disappearance were lower from 180 to 300 minutes during the meal versus control study (P < 0.05). Proteolysis was unaffected during the control study and was inhibited by 25% during the meal study (P < 0.05). During the five-hour cycle dialysis with or without a meal, 80% of the leucine administered into the peritoneum was absorbed. Forty-one percent was retained in the splanchnic bed. Forty-three percent was used for protein synthesis, and 16% was oxidized. CONCLUSIONS: This amino acids solution is efficaciously utilized for protein synthesis in CAPD patients with no effect on protein breakdown. The concomitant ingestion of a carbohydrate-lipid meal inhibits protein breakdown and reinforces a positive effect of the amino acids solution on protein balance.

Blood lipid concentrations of docosahexaenoic and arachidonic acids at birth determine their relative postnatal changes in term infants fed breast milk or formula.

BACKGROUND: Factors other than dietary fatty acids could be involved in the variability observed in blood docosahexaenoate (22:6n-3) and arachidonate (20:4n-6) status in formula-fed infants. OBJECTIVE: We considered the 22:6n-3 and 20:4n-6 status at birth to be one of these factors and studied its influence on postnatal changes in term infants fed 4 different diets. DESIGN: The blood phospholipid composition was determined at birth and on day 42 of feeding in 83 term infants fed breast milk, nonsupplemented formula, or 2 different 22:6n-3-supplemented formulas. Relations between 22:6n-3 and 20:4n-6 status at birth and their relative postnatal changes, calculated by the difference between status at the end of the feeding period (6 wk of age) and at birth, were assessed. RESULTS: Postnatal changes in the plasma and erythrocyte phospholipids 22:6n-3 and 20:4n-6 were negatively related to their respective concentrations at birth (P < 0.01) and the slopes of the regression lines were not significantly affected by the type of milk ingested. Adjusted mean values for phospholipid 22:6n-3 in nonsupplemented-formula-fed infants and for 20:4n-6 in formula-fed infants decreased significantly more than they did in the other infant groups (P < 0.02). The status at birth and the type of milk ingested explained 33-64% and 7-47%, respectively, of the variability in postnatal changes. CONCLUSIONS: The status of 22:6n-3 and 20:4n-6 at birth in term infants is one of the major determinants of postnatal changes in these fatty acids. This finding indicates that research is required to characterize environmental, genetic, or both factors, which, in addition to maternal diet, could influence fatty acid status at birth.

Intake of fatty acids in western Europe with emphasis on trans fatty acids: the TRANSFAIR Study.

OBJECTIVE: To assess the intake of trans fatty acids (TFA) and other fatty acids in 14 Western European countries. DESIGN AND SUBJECTS: A maximum of 100 foods per country were sampled and centrally analysed. Each country calculated the intake of individual trans and other fatty acids, clusters of fatty acids and total fat in adults and/or the total population using the best available national food consumption data set. RESULTS: A wide variation was observed in the intake of total fat and (clusters) of fatty acids in absolute amounts. The variation in proportion of energy derived from total fat and from clusters of fatty acids was less. Only in Finland, Italy, Norway and Portugal total fat did provide on average less than 35% of energy intake. Saturated fatty acids (SFA) provided on average between 10% and 19% of total energy intake, with the lowest contribution in most Mediterranean countries. TFA intake ranged from 0.5% (Greece, Italy) to 2.1% (Iceland) of energy intake among men and from 0.8% (Greece) to 1.9% among women (Iceland) (1.2-6.7 g/d and 1.7-4.1 g/d, respectively). The TFA intake was lowest in Mediterranean countries (0.5-0.8 en%) but was also below 1% of energy in Finland and Germany. Moderate intakes were seen in Belgium, The Netherlands, Norway and UK and highest intake in Iceland. Trans isomers of C18:1 were the most TFA in the diet. Monounsaturated fatty acids contributed 9-12% of mean daily energy intake (except for Greece, nearly 18%) and polyunsaturated fatty acids 3-7%. CONCLUSION: The current intake of TFA in most Western European countries does not appear to be a reason for major concern. In several countries a considerable proportion of energy was derived from SFA. It would therefore be prudent to reduce intake of all cholesterol-raising fatty acids, TFA included.

Fatty acid composition of white adipose tissue and breast milk of Mauritian and French mothers and erythrocyte phospholipids of their full-term breast-fed infants.

The fatty acid compositions of white adipose tissue, colostrum and mature milk triacylglycerols from Mauritian (n 13) and French (n 15) women were analysed and compared in order to highlight cultural differences in dietary intakes and their influence on milk fatty acid composition. Erythrocyte phosphatidylethanolamine and phosphatidylcholine fatty acid compositions were also investigated in their term infants, breast-fed over a period of 6 weeks. Fatty acid composition (g/100 g) of all samples was determined by GLC and anthropometric measurements were assessed in the two populations at birth and on day 42. Comparisons of white adipose tissue fatty acid compositions demonstrated lower levels of saturated (23.64 (SE 1.54) v. 29.75 (SE 0.67), P < 0.01) and monounsaturated (39.44 (SE 1.27) v. 54.84 (SE 0.75), P < 0.001) fatty acids and higher levels of polyunsaturated fatty acids (n-6 series: 32.47 (SE 1.31) v. 14.32 (SE 0.47), P < 0.001 and n-3 series: 2.87 (SE 0.49) v. 0.80 (SE 0.07), P < 0.01) in Mauritian than in French samples respectively. Accordingly, milk fat of the Mauritian women contained higher levels of parent essential fatty acids and their longer-chain derivatives than did milk fat from French women. Higher levels of parent essential fatty acids but lower levels of long-chain polyunsaturated fatty acids were found in erythrocyte phospholipids of Mauritian infants compared with French infants. Infants' erythrocyte arachidonate and docosahexaenoate contents did not correlate with any anthropometric variables at birth or at day 42, neither did they correlate with anthropometric variation over the study period. Our results suggest the lack of a simple relationship between the amount of long-chain polyunsaturated fatty acids in human milk and their accretion in the erythrocyte phospholipids of breast-fed infants when provided concomitantly with high levels of both linoleic and alpha-linolenic acids in ratios which fall within recommended ranges.

Effects of oral glucose on intermediary metabolism in continuous ambulatory peritoneal dialysis patients versus healthy subjects.

OBJECTIVE: To explore the mechanisms and metabolic consequences of the insulin resistance of patients treated by continuous ambulatory peritoneal dialysis (CAPD). DESIGN: CAPD patients and healthy subjects ingested a similar mean oral glucose load per kilogram of fat-free mass (FFM) [1.20+/-0.03 g/(kg FFM) vs 1.20+/-0.06 g/(kg FFM); CAPD vs healthy subjects]. Substrate oxidation was monitored over 6 hours using indirect calorimetry. SETTING: Peritoneal dialysis unit of a tertiary-care institutional center. OUTCOME MEASURES: Glycemia, insulinemia, substrate oxidation. PATIENTS: Six CAPD patients (68+/-5 yr) and 6 healthy subjects (24+/-1 yr). The CAPD patients had similar body mass index (21.4+/-1.3 vs 22.9+/-1.1 kg/m2), a higher percent fat (25.8%+/-3.7% vs 16%+/-2.2%; p < 0.05), and a lower FFM (42.2+/-2.2 kg vs 56.5+/-2.6 kg; p < 0.01) than healthy subjects. RESULTS: The CAPD patients displayed a higher glycemic and insulinemic responses to glucose than did healthy subjects (p < 0.05), but similar glucose oxidation and storage. Lipid oxidation and plasma nonesterified fatty acids were not increased in CAPD patients versus healthy subjects, in spite of a higher adiposity. Fat oxidation was related to fat mass in CAPD patients (r2 = 0.77, p < 0.05) but not in healthy subjects (r2 = 0.05). CONCLUSION: CAPD patients display an insulin-resistance not explained by an increased lipid oxidation. The maintenance of intracellular glucose utilization at the expense of higher glycemic and insulinemic responses suggests a defective glucose transport.

Effect of two types of fish oil supplementation on plasma and erythrocyte phospholipids in formula-fed term infants.

We studied the effect of docosahexaenoic acid (DHA) supplementation of infant formulas on fatty acid composition of blood phospholipids in term infants. Two fish oil supplemented formulas containing 0.45 wt% DHA and high (0.35%) or low (0.10%) eicosapentaenoic acid (EPA) were fed for 42 days and compared with a standard formula and breast milk. Infants fed supplemented formulas and breast milk had similar time-dependent changes for DHA from birth to day 42, i.e., slight decreases in plasma phospholipids and erythrocyte phosphatidylcholine and no change in erythrocyte phosphatidylethanolamine. Low-EPA formula prevented EPA accumulation but did not limit the significant decrease in arachidonic acid (AA) noted in infants fed high-EPA formula. These results suggest that term infant formulas should be supplemented with DHA-rich EPA, low fish oil and AA to achieve a fatty acid status in formula-fed infants similar to that of breast-fed infants.

Long-term supplementation of culture medium with essential fatty acids alters alpha-linolenic acid uptake in Caco-2 clone TC7.

We investigated the influence of four different culture media: 20% fetal bovine serum (FBS), 5% FBS, 5% FBS supplemented with 10 mg x L(-1) linoleic acid (18:2(n-6)) or alpha-linolenic acid (18:3(n-3)) on alpha-linolenic acid apical uptake in clone TC7 of human intestinal Caco-2 cell line. Neither cellular viability nor cell monolayer integrity and permeability were altered by the four culture conditions. Our results show that the different culture media led to changes in alpha-linolenic acid maximal rate of uptake (Vmax) but did not alter the apparent transport constant (Km). Reducing FBS concentration from 20% to 5% increased significantly the rate of alpha-linolenic acid uptake, which was further increased by supplementation of the medium with 18:2(n-6) or 18:3(n-3). Supplementation with essential fatty acids led to a marked enrichment of brush-border membrane phospholipids in polyunsaturated fatty acids of the corresponding series and decreased significantly the levels of monounsaturated fatty acids. Saturated fatty acids, unsaturation index, and cholesterol/fatty acid ratios were unchanged. No clear relation could be established between the changes in membrane lipid composition and the alterations of alpha-linolenic acid uptake. These results indicate a weak influence of membrane lipid composition in the modulation of the uptake. Therefore, the increase of uptake following long-term supplementation of TC7 cells with essential fatty acids could be attributed to an increase of the expression of membrane protein(s) involved in the apical uptake of long-chain fatty acids. This remains to be established.

Effect of dietary fish oil on body fat mass and basal fat oxidation in healthy adults.

OBJECTIVE: To investigate whether the substitution of fish oil for visible fats in a control diet (52% carbohydrates, 16% protein, 32% fat; P:S 0.2) influences body fat mass and substrate oxidation in healthy adults. DESIGN: Six volunteers (5 men; 23 +/- 2 y; BMI: 21.9 +/- 1.6) were fed a control diet (C) ad libitum during a period of three weeks and, 10-12 weeks later, the same diet where 6 g/d of visible fat were replaced by 6 g/d of fish oil (FO) for another three weeks. RESULTS: Energy intakes (IKA-calorimeter) were unchanged. Body fat mass (Dual-energy X-ray absorptiometry) decreased with FO (-0.88 +/- 0.16 vs -0.3 +/- 0.34 kg; FO vs C; P < 0.05). When adjusted for lean body mass (Ancova), resting metabolic rate (indirect calorimetry) was unchanged. Basal respiratory quotient decreased with FO (0.815 +/- 0.02 vs 0.834 +/- 0.02; P < 0.05) and basal lipid oxidation increased with FO (1.06 +/- 0.17 vs 0.87 +/- 0.13 mg kg(-1) min(-1); P < 0.05). CONCLUSION: Dietary FO reduces body fat mass and stimulates lipid oxidation in healthy adults.

Processing/activation of CPP32-like proteases is involved in transforming growth factor beta1-induced apoptosis in rat hepatocytes.

Apoptosis induced in rat hepatocytes by transforming growth factor beta1 (TGF-beta1) was accompanied by the activation of interleukin-1beta converting enzyme (ICE)-like proteases. Cell lysates were isolated at various times after TGF-beta1 treatment and analyzed for ICE and CPP32-like activity, using N-acetyl-Tyr-Val-Ala-Asp-7-amino-4-methylcoumarin (Ac-YVAD.AMC) and benzyloxycarbonyl-Asp-Glu-Val-Asp-7-amino-4-trifluoromethylcoumarin (Z-DEVD.AFC), respectively. CPP32-like but not ICE protease activity increased in a time dependent manner and preceded the onset of apoptosis. Kinetic studies in cell lysates indicated that more than one CPP32-like protease was being activated. This was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting of TGF-beta1-treated cells, which showed limited processing of CPP32 as shown by the appearance of the catalytically active p17 subunit. Loss of pro-Mch3alpha was also observed but the catalytically active p19 subunit was not detected. Staurosporine, which induced a much greater level of hepatocyte apoptosis, produced a concomitant increase in CPP32/Mch3alpha processing as shown by the appearance of the p17/p19 subunits and the corresponding increase in CPP32-like protease activity. Apoptosis, CPP32/Mch3alpha processing and the increase in CPP32-like protease activity induced by TGF-beta1 and staurosporine were abolished in hepatocytes pretreated with Z-Asp-Glu-Val-Asp (OMe) fluoromethylketone (Z-DEVD.FMK) or Z-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK). These peptide analogues were potent inhibitors of CPP32-like protease activity in lysates. Pretreatment of hepatocytes with cycloheximide also blocked TGF-beta1-induced apoptosis and the increase in CPP32-like activity. Unlike Z-VAD.FMK and Z-DEVD.FMK, cycloheximide did not inhibit CPP32-like protease activity in cell lysates. Thus, cycloheximide may block apoptosis by inhibiting the synthesis of a protein, which is involved in the upstream events responsible for the activation of the CPP32-like protease activity. Our studies have identified two of the CPP32-like proteases, namely CPP32 and Mch3alpha, which are activated during the execution phase of hepatocyte apoptosis.

Mechanisms and kinetics of alpha-linolenic acid uptake in Caco-2 clone TC7.

The uptake kinetics of alpha-linolenic acid (18:3(n - 3)), an essential fatty acid, were investigated in the human intestinal cell line Caco-2. Four clones (PD10, PF11, PD7 and TC7) from the heterogeneous parental Caco-2 cells population were used. After a screening step using isolated cells, the TC7 clone was selected for the study of alpha-linolenic acid uptake. [1-(14)C]linolenic acid dissolved in 10 mM taurocholate was presented to the microvillus plasma membrane (apical side) of TC7 differentiated cells, grown on a semi-permeable polycarbonate membrane. The results show that the initial rate of uptake is not a linear function of the 18:3(n- 3) monomer concentration in the incubation medium. In the monomer concentration range studied (0.2 to 36 microM) apical uptake was saturable and followed Michaelis-Menten kinetics (V(max) = 15.4 +/- 0.6 nmol/mg protein per min, K(m) = 14.3 +/- 1.3 microM). In addition, it was temperature- and energy-dependent but was apparently unaffected by the sodium gradient and intracellular metabolic fate of 18:3(n - 3). Excess of unlabeled saturated or unsaturated long chain fatty acids (C16 to C22) led to a 27-68% reduction of [1-(14)C]linolenic acid uptake. Likewise basolateral uptake was saturable (V(max) = 4.9 +/- 0.7 nmol/mg protein per min, K(m) = 8.7 +/- 2.9 microM). These facts argue in favour of the existence in these human intestinal cells of a carrier-mediated transport system for alpha-linolenic acid and probably other long chain fatty acids as well.