Method: U937 cells were incubated in media containing 0.05 or 2 mmol/IGLN supplemented with either 100 I!mol/I LA or 2 mmol/I NAG for 24 h.GSH content was measured by HPLC using monobromobimane for stain­ing.Results: GLN and NAG led to an enhancement of GSH levels in U937 cellswhereas the supply of LA had no effect under GLN deficient conditions.The combined administration of 100 I!M LA and 2 mmol/I GLN led to afurther (+100%) increase of intracellular GSH.

Results: The mean TPOAb concentrations decreases significantly to63.6% (p=0.013) in the selenium group versus 88% (p=0.95) in the place­bo group. Nine of verum versus 2 patients in the placebo group had nor­malized TPOAb levels (p=O.Ol). Ultrasound pattern were normalized inthese patients. The meanTSH, FT4 and FT3 values were unchanged in allpatients. Quality of life was signifi cantly (p < 0.001) improved in the sele­nium treated patients. The selenium concentration significantly increasedfrom 0.81 to l09 I!mol/I in the verum group and was unchanged in the pla­cebogroup.Conclusions: We conclude, that a selenium substitution improves the in­flammatory activity in patients with autoimmune thyroiditis as well as thequality of life. Whether these effects are specific for thyroiditis or may bealso effective in other autoimmune diseases has to be investigated.

wlosupplement+100 I!mol/I LA+2 mmol/I NAG

0.05 mmolll GLN

3.5 ± 0.63.5 ± 1.3°5.5 ± 2.9*

2 mmol/I GLN

6.3 + 2.4*12.6 ±3.0** ° ++

7.4 ± 3.7*

0-9: Stimulation of intracellular glutathione content by a-lipoicacid In the myelomonocytic cell line U937

E. Strasser, B. Wessner, A SpittlerandE. Roth

SurgicalResearchLaboratories,UniversityofVienna, GeneralHospital,

Vienna, Austria.

Rationale: The SH-donors ex-lipoic acid (LA) and N-acetylcysteine (NAG)are used as antioxidative substances under different clinical conditions. Inthis study we compared the stimulating effect of LA and NAG on glu­tathione (GSH) synthesis in dependence on glutamine (GLN) supply.

Intracellular GSH levels of U937 cells. Oata are expressed as nmol GSH/106 cells (mean ± SO; n=5). Significance by Mann-Whitney-test: **p < 0.01 vs 0.05 mM GLN w/o supplement; *p < 0.05 vs 0.05 mM GLN w/osupplement;op<0.05vs 2mM GLN w/o supplement; ++p<0.01vs0.05 mM GLN +100 I!M LA.Conclusions: In this experimental model, GSH content is dependent onGLN supply. But under conditions of GLN deficiency the administration ofNAG also enhanced intracellular GSH. Interestingly, the combined supplyof GLN and LA led to a remarkable increase of GSH levels in the myelomo­nocytic cell line U937. Therefore the concomitant administration of GLNand LA could be regarded as a powerful instrument in the treatment ofclinical conditions with lowered GSH levels and/or increased oxidativestress.

Oral Communications included in Scientific Symposia(1st Half of the Congress)

(please refer to programme for timings)

0-10; Tumor response, immunosurveillance and role of nutritionIn microgravity

M.Taga1, K. YamauchP, L. Furian1, J. Odle2, A Sandaresan2, N.R. Pellis3,R.J. AndrassiandAD. Kulkarni1

1Surgery,The UniversityofTexas HealthSienceCenterat Houston,2SpaceLifeSciences,USRA, 3CellularBiotechnologyProgram,NASA/JSC,Houston,UnitedStates.

Rationale: Microgravity (MG) induced immune dysfunction in space travelis a major health risk to humans and may endanger future long-term spacemissions.The incidence oftumorgrowth and carcinogenesis in micrograv­ity is yet unknown. Hence, we investigated the effects of simulated MG(SMG) on tumor growth and tumorigenecity using in vivo and in vitromodels.Method: Hence, we investigated the effects of simulated MG (SMG) on tu­mor growth and tumorigenecity using in vivo and in vitro models. B16 mel­anoma cells were cultured in static flask (FL) and rotating wall vesselbioreactors (BIO) to measure growth and properties, melanin productionand apoptosis.Results: BIO cultures had 50% decreased growth (p < 0.01), increaseddoubling time and a 150% increase in melanin production (p < 0.05). Flowcytometric analysis showed increased apoptosis in BIO. When BIO cul­tured melanoma cells were inoculated sc in mice there was a significantincrease in tumorigenecity as compared to FL cells. Thus SMG may havesupportedselected highly tumorigenic cells and it is possible that in addi­tion to decreased immune function MG may alter tumor cell characteristicsand invasiveness. In our previous unit gravity studies, nucleotide supple­mented diet significantly decreased growth and metastasis of melanomain mice.Conclusions: Immunomodulatory nucleotide nutrition enhances immunefunction in mice under SMG environments. Therefore, immunomodulatorynutrition may improve immunosurveillance and prevent carcinogenesis,tumor growth and metastasis in microgravity. Additional results will be pre­sented. (Supported by NASA NCG8-168 grant, AOK.)

3

0-11: Lymphocyte functions in mlcrogravity: selective signalinglesions in immune suppression and countermeasures

A. Sundaresan1 andN.R. Pellis2

1BiologicalSystemsOffice,UniversitiesSpace ResearchAssociation/NASA JohnsonSpace Center, Houston,UnitedStates, 2BiologicalSystemsOffice,NASA JohnsonSpace Center, Houston,USA

Rationale: Previous experiments by our laboratory demonstrated that lym­phocyte activation and locomotion are impaired in microgravity and mod­eled microgravity (mmg). Studies were performed to dissect cell signalingpathways and design countermeasures.Method: Immunoprecipitation and immunoblotting for activation and ex­pression of PLG gamma1, IP3 Rand PKG isoforms. RT-PGR to analysemessage of PKG isoforms. Locomotion assay for locomotion.Resuhs: Lymphocyte locomotion and activation were inhibited in mmg by100% (p < 0.0001). PMA could restore locomotion by 87% (p < 0.001) andactivation by 100% (p < 0.001).lonomycin had no restorative effects. SincePMA directly activates PKG, Ga++ independent PKG isoforms were evalu­ated. PKG d mRNA in mmg was down-regulated to almost invisible at 24and 48 H cultures and by 56% at 96 H. Protein expression was similar.PKG e had the same trend with an 83% decrease in message levels at 24and 48 h in mmg and a 53% decrease at 96 H. Protein levels were negligi­ble at 96 H in mmg. Activation of PLG g1 which is upstream of PKG was in­hibited by more than 2 fold (p < 0.05). Microarray analysis revealed down­regulation of activation genes such as OAG kinase (4.3 fold), human ser­ine/threonine kinase (3.8), tyrosine kinase (2.4).Conclusions: Optimal immune function is critical in the ISS era where longlerm space travel is inevitable. Elucidation of pathways affected and de­sign of nutritional countermeasures are of paramount importance. PMAcannot be administered to human SUbjects, and alternatives such as syn­thetic OAGs and nucleotides are being evaluated in locomotion studies inour laboratory. These would be crucial in microgravity, immunology andnutritional research.

0-12: A proteomics approach to Wilson disease: effect of copperon protein expression in an in vitro model of copper overload

R.J.Vonk', L. Lomas2 andH. Roelofsen'

'Pediatrics,CenterforLiver,DigestiveandMetabolicDiseases,Groningen,Netherlands,2ResearchandDevelopment,CiphergenBiosystems,Fremont,USA.

Rationale: In Wilson disease, mutations in the ATP7B-gene lead to hepaticaccumulation of copper that will cause hepatitis and acute liver failure.Several proteins are probably involved in dealing with the excess copperand the copper-induced oxidative stress. Using proteomics we character­ized copper-induced changes in protein expression in human HepG2hepatoma cells.Method: HepG2 cells were cultured for 24 hrs: (1) in the absence of copper;(2) With 0.12 11M Cu 2 +; or (3) with 100 11M Cu2+. Protein expression profi leswere obtained from cell Iysates using the SELDI technology (CiphergenBiosystems). This method uses ProteinChip'"' Arrays with different affi nitysurfaces to enhance for the detection of proteins with specific biochemicalcharacteristics. Ceillysates were applied to the arrays and, after washing,were analyzed by time of fl ight mass spectrometry.Results: Preliminary data from the 0.5 to 85 kD range show four proteins of5.8, 6,1, 39.6 and 50.8 kD that are clearly differentially regulated on in­creased extracellular copper, Already 0,12 11M Cu 2+ down-regulates the5.8 kD protein while it up-regulates the 6.1 kD protein. These effects areeven more pronounced with 100 11M CuN,The 39,6 and 50.8 kD protein be­come up-regulated only with 100 11M Cu2

+. Further identification is neces­sary to determine their exact cellular roles.Conclusions: Results show that already small changes in extracellularcopper concentration lead to alterations in protein expression, indicatingthat HepG2 cells provide a good human in vitro model to study effects ofcopper (overload) on hepatic protein expression.

0-13: Proteome analysis of the effect of glutamine depletion onmonocytic cells: potential involvement of the AMP-activatedkinase pathway

M.M. Eliasen, J. Pollheimer,M Zellner, A. Spittler,E. RothandR. Oehler

SurgicalResearchLaboratories,UniversityofVienna,Vienna, Austria.

Rationale: We investigated the consequences of reduced glutamine (Gin)levels on the protein expression profi Ie in monocytic U937 cells and char­acterised the role ot the AMP-activated kinase (AMPK) pathway in thisstarvation response.Method: Cells were cultured at reduced Gin levels for 4 d and analysed bytwo dimensional gel electrophoresis. This allows to evaluate the expres­sion of the majority of all proteins simultaneously, separating them accord­ing to their isoelectric point (PI) and molecular weight (MW).Results: Approx. 600 spots of proteins with PI between 3.0? 10.0 and MWfrom 25 kDa? 250 kDa were resolved. The majority of these proteins re­mained unaffected by Gin-depletion. We characterised 27 new proteinspots in Gin-starved cells, whereas 30 protein spots were completely re­pressed. Moreover, 15 protein spots showed a 10 to 100-fold increased ex­pression by Gin starvation, whereas 10 were 2 to 5-fold reduced. Monocyticcells utilise Gin as an energy source. Accordingly, the ATP level was re­duced by 33.8 ± 5.6% (p=0.004) in starved cells. Such an impaired cellularenergy charge leads to an activation of the AMPK pathway in several mam­malian cell types. Western blot analysis revealed a change in expressionand phosphorylation of AMPK also in Gin-starved monocytic cells.Conclusions: This study shows that Gin depletion induces a comprehen­sive starvation response in monocytic cells. This includes altered expres­sion of the activated AMPK. Since AMPK is known to modulate theexpression of several proteins involved in the energy metabolism we hy­pothesise that the shown changes in the protein expression profile areconnected to the AMPK-pathway.

0-14: Short-term moderate caloric restriction improves vascularfunction in lean rat aorta

M, Zanetti', R. Barazzoni',M Vador;2,M Stebel3 and G. Guarnieri4

'DepartmentofClinical,MorphologicalandTechnologicalSciences,ClinicaMedica, UniversityofTrieste,2DepartmentofBiomedicalSciences,InstituteofPharmacology,3CSPA, AnimalFacility,4DepartmentofBiomedicalSciences,ClinicaMedica, UniversityofTrieste,Trieste, Italy.

4

Rationale: Caloric restriction is associated with reduced cardiovascularrisk also in the absence of obesity. The molecular mechanism(s) of thisbeneficial effect are not understood. Endothelial relaxation in aorta is in­duced by nitric oxide (NO) produced by endothelial NO synthase (eNOS).Endothelial dysfunction is an early marker of atherosclerosis. We studiedthe effects of moderate caloric restriction on endothelial function in a leanrat model in vivo.Method: Six-month old Fisher-344 rats received either a standard ba­lanced chow diet ad libitum with monitoring of food intake (Control, n=8,initial body weight 372 ±8 g), or a 25% reduced amount of the same diet(CR, n=8, initial body weight 367 ±11 g) for one week. Aortas from Controland CR were then harvested and endothelial function was assessed byWestern analysis of eNOS protein levels and by isometric aortic tensionrecording.Results: CR lost weight compared to Control (-2.6 ± 0.2 vs. 1.1 ±0.4 g/d,P < 0.05). CR resulted in increased (+25 ± 2.1 %, P < 0.05) eNOS expres­sion in rat aorta. Acetylcholine (10-6 -3 x 10-6 and 10-5 M) mediated, en­dothelium-dependent vasorelaxation was higher (P<0.05) in CR(46.1 ±2.9, 58.5 ± 5.6, 68.9 ± 5.6%) than in Control (34.8 ± 4.1, 40.9 ± 3.9and 51.1 ±5.3%),Conclusions: These fi ndings demonstrate that short-term moderate calo­ric restriction improves endothelial function in lean rats by increasing theexpression of eNOS. These changes indicate a nutritional regulation of en­dothelial function and suggest a potential molecular mechanism for thebenefits of caloric restriction on cardiovascular risk.

0-15: Prediction of final stature in obese children andadolescents: a different growth pattern?

C. Rego', A. Guerra1, D. Silva2, S. Sinde2, M Fontoura'andA. Aguiar'

'Paediatrics,HospitalS. JoaoI FacultyofMedicineofPorto, 2paediatrics,HospitalS. Joao, Porto, Portugal.

Rationale: To calculate fi nal stature (FS) of pre and puber obese childrenand adolescents and to study the possible association between some hor­monal and somatic markers.Method: 83 subjects regularly followed in Nutrition Outpatient PaediatricClinic. At the 1stobservation, height, weight, body composition (bioelectri­cal impedance), bone age (BA) (Grewlich and Pyle) and sexual maturation(Tanner) were evaluated. Body mass index (BMI) and statural prediction(SP) (Baileyet Pinneau) were calculated. Gh, IGF1, fasting insulin and fast­ing glucose were measured and HOMA insulin resistance (IR) was calcu­lated. The sample was divided into two groups according to chronologicalage (CA): Group A <10 years (n=41) and Group B= > 10 years (n=42).Results: The mean CA was 10.3 ± 3.1 years. Group A showed an earlier on­set of obesity (p < 0.01), a greater body mass (Zs-BMI) (p < 0.001), a higherstature (p < 0.001) but no significant differences were observed regardingSP. When considering the whole sample, BMI values grouped by quartilesshowed higher values on those with higher fasting insulinemia (p < 0.05)and HOMA IR (p < 0.05) and those with lower values of IGF1 (p < 0.05). Asignificant positive correlation was observed between biological maturity(BA-CA) and stature (p < 0.05) in both genders. A significant positive cor­relation was also observed between BA-CA and Zs-BMI (p < 0.01) andfasting insulinemia (p < 0.05).Conclusions: Obesity is associated with a higher stature and with an accel­eration of biological maturity only during the prepubertal period comparedto puber individuals. Obesity seems to induce acceleration on heightvelo­city during prepubertal age, followed by a deceleration during puberty.

0-16: Relation of leptin components to sympathetic activity andhepatic free faUy acid metabolism in cirrhosis

J. Ockenga', U. Ttetge2, G. Brabant3 andMP. Manns2

'Gastroenterology,Hepatology,Endocrinology,HumboldtUniversity,Charite,Berlin,2Gastroenterology,3Endokrinology,MedicalSchoolHannover, Hannover,Germany.

Rationale: Leptin circulates in a free (FL) and a bound form (BL), which isformed by binding of FL to soluble leptin receptor (sLR). We recently found,that FL and BL have different effects in liver cirrhosis [Gastroenterology2000]. We investigated the relationship between the leptin componentsand hepatic substrate metabolism as well as sympathetic activity or hor­monal signals.

Adults Adults Elderly Elderlybefore after before afterinsulin insulin insulin insulin

Insulin receptor density 4456±238 4196+269 4351 ±383 4007 + 172Chemotaxis (fMLP/SM) 3.1 ±O.4a 6.7 ± 19b 3.8±0.5a 6.2 ± lObPhagocytosis (AU) 5.4 ± 1.0a 7.0 + l2b 3.2±0.3c 3.5+0.4c

ROS(AU) 203±33a 238±35b 86 ± 11c 111 ± 18d

Mean (standard deviation) biochemical values for individuals administered supple­ment, 'p< 0.01, "p=0.15, paired t-test. Placebo did not significantly affect biochem­ical variables over the 8-week study period.Conclusions: Supplementation given over 4 weeks significantlyincreased the level of vitamins A, C, E, selenium and folate andreduced the frequency of abnormalities. No significant effect wasseen for zinc; after 8 weeks of supplementation almost two thirds ofthis sample had zinc levels that remained below the lower limit of normal«11 umol/I).

ANOVA+Newman-Keuls a#b#c with p<0.05 AU: arbitrary unit fMLPISM: ratio of migration toward fMLP to spontaneous migration.Conclusions: We confirmed the presence of insulin receptors on PMNmembranes revealing a functional role of insulin on PMNs as indicated bythe stimulation of chemotaxis, phagocytosis and ROS production in youngsubjects. However, we clearly showed that phagocytosis is not responsiveto insulin in elderly people. This specific alteration in PMNs response toinsulin may significantly contribute to the lack of host defences in thispopulation.

matched placebo, n=58, as a tablet twice daily for 8 weeks. Venous bloodsamples were obtained at week 0, week +4 and week +8.Results:% S group below normal range at week 0:vit A 0%, vit C 65%, vit E 7.3%, zinc 62.7%, selenium 3.4%, folate,3.6%% S group below normal range at week +4:vitAO%, vit Cl8%, vit E 0%, zinc 62.7%, selenium 1.7%, folate 0%% S group below normal range at week +8:vitA 0%, vit C 3.5%, vit E 0%, zinc62.7%, selenium 1.7%, folate 0%.

No linear association was seen at week 0 between zinc levels and C­reactive protein (Spearman correlation cofficient, r=-0.182, p=0.05).

Folate(ug/l)

6.1 (4.1)12.0 (6.5)"

Se(umol/l)

Zn(umol/l)

VitE(umoi/l)

25.4 (8.8) 10.3 (1.6) 1.0 (0.2)36.1 (13.5)' 10.6 (2.0)" 1.1 (0.2)"

Vite(umol/l)

VitA(umol/l)

Week 0 2.9 (0.9) 23.6 (22.0)Week +4 3.1 (0.9)" 64.1 (20.5)'

0-19: Insulin differently affects neutrophil functions in healthyelderly and adult humans

S.Walrand1, C. GUillef, L. Morin2,Y Boirie2 andM. Vasson1

lLab Bioch,BioiMol et Nut"Fac de Pharma, 2UMPE, GRNH, Glermont­Ferrand, France.

Rationale: Since aging is associated with various degrees of insulin resis­tance in conjunction with reduced neutrophil functions, we investigated thehypothesis that neutrophils may be less responsive to insulin action in nonmalnourished elderly subjects.Method: We evaluated the effect of hyperinsulinemia (0.51mU.kg-1 min \euglycemia and hyperaminoacidemia clamp for 4 hours on PMNs functionin 7 young and 8 elderly healthy volunteers. Blood was withdrawn forPMNs isolation before and after the: clamp. Insulin receptor density wasmeasured using fluorescent antibody and flow cytometric detection.PMN chemotaxis was evaluated toward formyl-Met-Leu-Phe using a twocompartment chamber. PMN phagocytosis and microbicidal function weredetermined by measuring the engulfement of opsonized particules, andthe reactive oxygen species (ROS) production of phorbol-myristate-acet­ate stimulated PMNs, respectivelyResults:

Method: Using arterial and hepatic venous catheterisation technique weinvestigated pre- and post-hepatic serum concentrations of FL, BL, andsLR, catecholamines, insulin, glucagon and free fatty acids (FFA) in 39cir­rhotics.Results: FL did not differ in cirrhotic and control subjects and was related tothe fat mass (cirrhotics: r=0.45, p < 0.01). There was no significant differ­ence between pre- and posthepatic concentrations of FL, BL or sLR. BLand sLR were significantly elevated in cirrhotic subjects compared to con­trols (0.73 ± 0.52 vs. 0.54 ± 0.29; 17.75 ± 22.9 vs. 5.06 ± 2.3p < 0.001) andshowed a close relationship to one another (r=O.7, p < 0.001). SLR concen­trations were associated to the catecholamines (noradrenaline r=0.45;adrenaline r=0.56; dopamine r=0.52, p<0.001), whereas FL correlatedwith insulin (r=0.518, p< 0.001) and FFA clearance of the liver (r= 0.48,p<0.003).Conclusions: Free leptin reflects fat mass and is related to hepatic FFAclearance and, thereby, may contribute to the development ofsteatosis. The increase in sLR is responsible for the increased circulatingBL, which is formed from FL binding to sLR. In addition, our data provideevidence, that sLR and BL are involved in the regulating of sympatheticoutflow, which may be responsible for inadequate energy expenditure incirrhotics.

0-17: Does leptin have a role in the response to feeding incachectic cancer patients?

M.D. Barberi, K.G.H. Fearon1, D.C. McMillan2, M. Wallace3 andT Preston4

ISurgery,RoyalInfirmary,Edinburgh,2Surgery,3Biochemistry,RoyalInfirmary,Glasgow,41sotopeBiochemistry,SUERC, EastKilbride,UnitedKingdom.

Rationale: Many cachectic cancer patients are hypermetabolic despite lowleptin levels. This study aimed to examine the relationship between leptinand other mediators and fat oxidation during feeding.Method: After an overnight fast, 7 weight-losing pancreatic cancerpatients and 6 healthy controls underwent measurement of circulatingleptin, interleukin-6, cortisol and insulin concentrations and fatoxidation over a 4 h feeding period. Data are presented as median(range) and statistical analysis was performed using non-parametrictests.Results: At baseline, the cancer group had lower leptin (2.1 (l3-14.6) ver­sus 12.9 (5.1-49.8) ug/I, P< 0.05) concentrations but higher interleukin-6 (4.1«2-9.4) versus <2 «2-< 2) UII, p<0.05) and cortisol (639 (437-677)versus 521 (382-626) nmol/I, p < 0.05) concentrations compared with con­trols. Leptin concentrations in the cancer group remained lower than con­trols when adjusted for fat mass (p < 0.05). Resti ng energy expenditure andfat oxidation were greater in the cancer group (p < 0.05). On feeding, therewere no significant changes in leptin or interleukin-6 concentrations ineither group. Cortisol concentrations fell (p < 0.01) and insulin levels rose(p<0.01) with feeding in both groups. Fat oxidation fell in the cancer pa­tients (p < 0.05). In the cancer patients the area under the curve of leptinconcentration significantly correlated with that of cortisol (r=-0.857,p<0.05).Conclusions: Leptin does not appear to playa major role in the fat meta­bolic response to feeding in weight-losing cancer patients or controls. In­terleukin-6 may upregulate the cortisol response to feeding in cancerpatients promoting fat oxidation despite limited stores.

0-18: The effect of micronutrient supplementation in olderinstitutionalised people

S.J. Allsupl, M. Gosneyl,B. Taylo?,M. Hammond1, S. Taylor3andA. Shenkin2

lGeriatricMedicine,2ClinicalGhemistry,3MathematicalSciences,Universityof LiverpOOl,LiverpOOl,UnitedKingdom.

Rationale: We investigated whether a supplement providing thereference nutrient intake (RNI) for vitamins and trace elements could re­verse micronutrient deficiencies thought to be common in institutionalisedolder people.Method: 119 residents (median age 82.6 y) of nursing or residential homesin Liverpool were randomised to receive either supplement (S), n=61, or

5

0-20: Mitochondrial protein synthesis and enzyme activities aredifferently affected by meal intake and leucine supplementation

C. GUilletl, P. RoussetI, C. GiraudetI, C. Sornet2, D. Dardevet2and Y. Boiriel

Iproteino-energeticMetabolismUnit, 2NutritionandProteinMetabolismUnit, CRNH-Auvergne,Clermont-Ferrand,France.

Rationale: Energy production in muscle results from substrates oxidation inmitochondria through numerous enzymes activity. Changes in substratesavailability during feeding potentially regulate mitochondria functions andprotein synthesis but this effect may be blunted during aging and pre­vented by leucine.Method: Adult (A=8 mo old) and old (0=22 mo old) rats were studied eitherin postabsorptive state ( PAl or after a meal (PP) or a leucine-supplemen­ted meal (PP+Leu). Fractional (FSR) synthesis rate of mitochondrial (mito)proteins was measured, in gastrocnemius muscle, 150 min after meal dis­tribution using a flooding dose of [1_13C] Phenylalanine. Maximal enzymeactivities of citrate synthase (CS) and cytochrome c oxidase (COX) wereassessed in muscle homogenate by photospectrometric analyses.Results: Mito FSR was not different in PA, PP or PP+Leu in A and 0. In PA,CS was lower in 0 than in A (234±14vs 265±23llmol/min/gprot;P<0.0001,Ovs A). In PP,CS and COX were stimulated (+15% and +21 %;P < 0.05, PP vs PA, respectively) in A but not in 0. In PP+Leu, CS did notdiffer from PA in A and O. COX was stimulated by leucine in both groups(A: 126 ±28 vs 100 ±12llmol/min/gprot; P<0.05, PP+Leu vs PA and 0:136 ±25 vs 105 ±15llmol/min/g prot; P<0.01, PP+Leu vs PAl.Conclusions: Meal intake stimulates mitochondrial enzyme activities inyoung not in old rats but leucine supplementation normalizes the effect ofmeal on mitochondrial functions. These age-related alterations in re­sponse to feeding are to be considered in situation of an increased energydemand during sepsis, trauma or stress.

0-21: Glutathione peroxidase gene knockout accelerates liverregeneration after partial hepatectomy

N. Iwakuma, N. Ishibashi,K. Momosaki,S. YoshidaandK. Shirouzu

0-22: A polymorphism at -511 in the IL-1 pgene is associated withdifferences in the metabolic response to surgery in cancerpatients

A. Thorell I, P. MaIF, G. Burdge2, G. 0'Reilly2, J. Nygrenl, O. Ljungqvistl and

R. Grimble2

ICentreofGastrointestinalDisease, ErstaHospital,Stockholm,Sweden,21nstituteofHumanNutrition,UniversityofSouthampton,Southampton,UK.

Rationale: Profound metabolic changes follow surgery, due to release ofthe pro-inflammatory cytokines IL-113.IL-6 and TNF-a:. Changes in aminoacid metabolism are reflected in alterations in urinary excretion of end­products. We have shown a decrease in the inorganic sulphate to nitrogenratio in urine following surgery that may reflect enhanced retention of sul­phur amino acids due to increased acute phase protein and glutathionesynthesis. The CTaliele combination at -511 in the promoter of the IL-1 pgene has been shown to be associated with enhanced inflammatorystress, as for example in IBD patients and alcoholic patients developingcirrhosis.Method: Total nitrogen and inorganic sulphate excretion was measuredpre- and for 5 d post-operatively in 39 patients undergoing surgery forG-I cancer. Patients were genotyped for the polymorphism at ~511 in thepromoter ofthe IL-1 pgene.Results: There was a significant fall in the sulphate to nitrogen ratio, frompre-operative values, at days 2-5 post-operatively. Lowest values were atd3. (1.7 fold fall, p < 0.0001 ANOVA). Genotype infl uenced the extent of thefall in sulphate excretion. The area under the excretion curve for individualswho were CC was greater (p<0.01 ANOYA) than those who had the CTallele combination (indicating a smaller fall in CC than CT individuals).There were no differences in age, gender or type of surgery between thetwo groups.Conclusions: The greater fall in sulphate excretion in CT than CC indivi­duals in the present study support the hypothesis that that the CT allelecombination at -511 in the promoter of the IL-1 p gene is associated withenhanced infl ammatory stress in patients undergoing surgery for gastro­intestinal cancer.

Serum AST levels, RLR and BrdUlabelingindicesafterpartialhepatect­omyin each group.Conclusions: Lack of GPx1 caused an increase of liver regeneration afterpartial hepatectomy without harmful effect on remnant liver.

DepartmentofSurgery,KurumeUniversitySchoolofMedicine,FUkuoka,Japan.

Rationale: Reactive oxygen species and antioxidant enzymes have beenimplicated in control mechanisms of cellular growth and proliferation.There is an accumulating evidence that supplementation of antioxidantas well as antioxidant enzymes gene transfer reduce liver injury, resultsimprovement of liver regeneration after partial hepatectomy, however, noevidence can be seen in the literature using selective antioxidant enzymesgene knockout model for this study to date. Here, we investigate the infl u­ence of gene knockout of intra-cellular glutathione peroxidase (GPx1) onliver regeneration.Method: C56BLl6 background normal mice (wild group) and GPx1 knock­out mice (KO group) were underwent a two-third partial hepatectomy (PH).48 hr after PH, mice were sacrificed, remnant liver and blood were har­vested. Serum levels of AST, rate of liver regeneration (RLR) and BrdU la­beling index were measured.Results: Data were expressed as mean (SEM). *: p < 0.05 compared to KOgroup.

AST levels (SF Units/ml)Rate of the liver regeneration (RLR)BrdU labeling index (%)

Wild group(n=6)

206 (72)*2.4 (0.1)*7.8 (0.1)*

KOgroup(n=6)

76 (9)3.4(0.3)

13.6(0.2)

6

0-23: Cytokine genotypes are related to some metabolic andclinical features of critical illness

C.L. Reidl andI. I Campbel/2

IDepartmentNutrition& Dietetics,CharlngCrossHospital,London,2DepartmentofAnaesthesia,WithingtonHospital,Manchester,UnitedKingdom.

Rationale: We have previously shown that systemic cytokine concentra­tions are statistically significantly related to metabolic features of criticalillness (ESPEN,1999). However short circulating half-lives of these media­tors can mean the true extent of the cytokine response may be missed.We therefore investigated relationships between cytokine genotypes andmetabolic and clinical features of critical illness (CI).Method: Cytokine genotypes (TNFa, TNFI3. IL-10 and TGFP1) were deter­mined in 88 ICU patients. Energy expenditure (EE) and muscle wastingwere measure daily. Urinary nitrogen (UN), cortisol and CRP levels weremeasured on alternate days (5-30 d). Severity of illness was scored dailyusingTISS and MOD scores. One-way ANOVAwas used to analyse differ­ences between genotype groups.Results: Mean cortisol levels were signi fi cantly higher in high IL-10 produ­cers compared with low or intermediate producers (1088.81 nmol/l vs.466.3 nmol/l and 463.0 nmol/I respectively; p=0.001). High TGFp1 produ­cers tended to need fewer days of venti latory support compared with lowand intermediate producers (5.8 d vS. 8.4 and 8.2 d, respectively; p=0.137)and they had fewer days with > 1 organ failure (4.8d vS. 7.4 and 7.5 d re­spectively; p=O.077).TNFa: genotypes showed no significant relationshipswith metabolic features of Cl.ln relation toTNFp genotype, CRP concentra­tions were significantly higher in B2 homozygotes compared with eitherB1 homozygotes or heterozygotes (189.8 mg/lvs.111.5 mg/l and 149.5 mg/lrespectively; p=0.026).Conclusions: Relationships between cytokine genotypes and the meta­bolic and clinical features of CI are not consistent with relationships re­ported previously using circulating levels of these mediators.

0-24: Effects of n3 -fatly acids plus oleic acid supplemented milkon CV risk factors in young healthy volunteers

L. Baro, J. Fonolla,J.L. Pefia, A Martinez-Ferez,J.J. Carrero,J. Jimenez,J.J. Boza andE. Lopez-Huertas

Nutrition,Puleva BiotechS.A, Granada, Spain.

Rationale: Numerous studies suggest beneficial effects on CVD due to ol­ive oil and n-3 fatty acids (EPA+DHA) consumption.This study asseses theeffect of a dairy product supplemented with an oil mix enriched in n-3 fattyacids and oleic acid plus folic acid and B-complex vitamins, on some riskfactors of cardiovascular disease.Method: Thirty healthy normolipidaemic volunteers (25-45 y) were fi rst gi­ven 0.51/day of semi-skimmed milk for 4 weeks and then 0.51/day of Pulevaw3 for 8 further weeks. Plasma and LDL lipoproteins were obtained fromvolunteers at the beginning of the study (TO) and at 4, 8 and 12 weeks. Re­levant biomarkers related to lipid metabolism and CVD were measured inplasma and LDL.Results: Results from this study show 2 months administration of Pulevaw3 produces a significant decrease in total (4.56 ± 0.11 vs 4.30 ±0.11 mmol/L; 0) and LDL cholesterol levels (2.35 ± 0.12 vs 1.97 ± 0.11 mmol/L; 0), increased plasma concentrations of Vit B12 (380 ±21 vs 474±20 ng/L; oJ, increased plasma (3.18 ± 0.26 vs 6.10 ± 0.45 ~g/L; 0) and erythrocyte(354 ±26 vs 436±23 ~g/L; 0) concentrations of folic acid accompanied bya significant reduction in plasma concentrations of homocysteine (12.5 ±3.4 vs 10.9 ± 3.0 ~g/L; 0). LDL oxidability, measured as lag time (155.3 ±8.5 vs 198.2 ±15.6 min; 0) and basal hidroperoxides (19.8 ±1.1 vs16.7 ±0.6 ~M/mg LDL prot; 0), was significantly reduced. Endothelial acti­vation markers were also measured and a significant reduction in plasmaconcentrations of VCAM-1 (10.2 ± 0.6 vs 8.6 ± 0.59 ~g/L; 0) was found.'p< 0.05Conclusions: Daily intake of n-3 and oleic acid supplemented skimmedmilk plus folic acid and B-complex vitamins reduces some risk factors forcardiovascular disease.

0-25: A randomised controlled study of vitamin C & Esupplementation following acute ischaemic stroke

R. Ullegaddi andS.E. Gariballa

SheffieldInstitutefor StudiesonAgeing,UniversityofSheffield, Sheffield,UnitedKingdom.

Rationale: Enhanced antioxidant capacity after acute ischaemic strokemay be protective against the adverse effect of free radicals productionduring ischaemia and reperfusion. The aim of this study was, therefore, todetermine if supplementation of acute ischaemic stroke patients with vita­mins C and E within a defined period of time post-infarct improves antiox­idant capacity.Method: Twenty ischaemic stroke patients were randomly assigned to re­ceive 800 IU (727 mg) of vitamin E and 500 mg of vitamin C orally or notreatment within 12 h of stroke onset, and then once daily for 14 days. Pa­tients in the two groups were matched for clinical stroke subtype and age«75, >75 y). Non-fasting venous blood was obtained at baseline, 7 and 14days post-randomisation for measurements of total antioxidant capacity(TAOC), cholesterol-adjusted vitamin E and vitamin C concentrations. Cu­mulative changes in plasma concentrations were compared between thetwo groups using the Kruskal-Wallis test.Results: The table shows the mean changes in plasma concentrations ofVitamin E,Vitamin C and TAOC over days 0-7,7-14 and 0-14. P-values forthe difference in cumulative changes between treatment and controlgroups were p=0.009, p=0.034, and p=0.021 for vitamin E, vitamin C andTAOC respectively.

GROUP VitE VitC TAOCJ,lmol/mmol Chol J,lmol/L mmol/L

Day Day Day Day Day Day Day Day Day0-7 7-14 0-14 0-7 7-14 0-14 0-7 7-14 0-14

Treatment 4.75 0.70 5.81 51.52 -26.32 25.20 0.15 -0.05 0.09(n=10),7femalesCONTROL 0.91 -0.20 0.30 -70.42 0.87 -69.55 0.D1 -om -0.06(n=10),6females

Conclusions: Supplementation of acute ischaemic stroke patients with 800IU of vitamin E and 500 mg of vitamin e for 2 weeks significantly increasedplasma concentrations of vitamins C and E and TAOe. Further studies areneeded to evaluate clinical benefits of such simple treatments in this groupof patients.

Oral Communications included in Education and Clinical Practice Symposia(1st Half of the Congress)

(please refer to programme for timings)

0-26: Enteral nutrition in critically ill patients with severehemodynamic failure.

M.M. Berger, M.C. CayeuxJ. RevellyandR. Chiolero

SurgicallCU,CHUV, Lausanne,Switzerland.

Rationale: Patients with circulatory failure are considered difficult to feedby the enteral route. With high levels of vasopressors, enteral nutrition (EN)is commonly considered contraindicated. There are no data for guidance,only expert opinion.This stUdy assessed energy delivery by enteral route inthis category of critically ill patients.Method: Prospective observational stUdy in 70 consecutive patients aftercardiac surgery with extracorporeal circulation. Inclusion criteria: ICU stay> 5 d and acute organ failure. Energy target set 25 kcal/kg/d to be reachedstep-by-step over 5 days (0%,0%,25%,50%,75%,100%). Enteral routewas considered fi rst. Variables: energy delivery, route, vasopressors,complications. Organ failure was assessed with the SOFA score.Results: 70 patients aged 67 ± 17 yrs were studied (mean ± SD). Length ofICU stay was 10 ± 7 d. Median SAPS II score was 43: 9 patients died (13 %).All patients stayed due to cardiac or respiratory complications (severe re­spiratory failure in 59 patients; severe circulatory failure in 58 patients -17patients required intra-aortic balloon-pump support (IABP)). Noradrena­line was required in 58 patients. Forty patients required artificial nutrition,TPN was combined with EN in only 3 patients. Individual and daily energydelivery was very variable. As a mean, 1360 ± 620 kcal/kg could be deliv-

ered by enteral route during the fi rst 2 weeks (70 ± 35% of energy target).Enteral energy delivery was not correlated with either noradrenaline dose,or use of IABP.There was no severe abdominal complication related to ENConclusions: EN is possible in many patients with severe hemodynamicfailure, but usually results in hypocaloric feeding. Considering these data,EN should be considered in patients with circulatory failure with carefulabdominal and energy monitoring

0-27: Randomized controlled trial on nutritional support in COPO,during hospitalization for an acute exacerbation

M.A.P.Vermeeren1, E.F. Wouters2, AJ. Geraerts3 andAM. Schols2

10ieteticsandPulmonology,2Pulmonology,UniversityHospitalMaastricht,Maastricht,30ieteticsandPulmonology,Maxima MedicalCenter,Veldhoven,Netherlands.

Rationale: To investigate the feasibility of oral nutritional supplementationto improve energy balance in depleted COPD patients hospitalized for anacute exacerbation.Method: Randomized controlled trial. Inclusion criteria: Body Mass Index(BMI)=22kg/m2 or BMI<25kg/m2 with >5% weight loss. Intervention(I): 3 x 125 ml Respifor,1\ (2.38 MJ/d; 20 energy% protein, 20 energy% fatand 60 energy% CHO). Placebo (P): 3 x 125 ml vanilla flavored water (0MJ/d). Medical therapy and dietetic consultation was standardized. Dietaryintake and body weight were recorded daily.