Thyroid function and metabolic risk factors in obese youth. Changes during follow-up: a preventive mechanism?

OBJECTIVES: High TSH levels often observed in overweight subjects are associated with metabolic risk. Thyroid hormones which are involved in fat and carbohydrates metabolism are more rarely studied; their blood levels were measured to more precisely explain the relationships between thyroid function and obesity, in healthy overweight youth. This correlation was studied at baseline and during follow-up of some patients. MATERIALS/METHODS: Data collected were BMI and BMI z score, thyroid hormones (TSH, fT4, fT3), fasting blood glucose, HOMA-IR, lipids (triglycerides, HDL and LDL cholesterol), transaminase activity, fibrinogen, leptin, IGF-I; body composition (biphotonic absorptiometry). Data collected in a sample of the group after 6-18 months of medical intervention could also be studied. RESULTS: At baseline, 13% of the 528 obese subjects (55% girls; 11.3±2.4 years, range 4.1-17.9; BMI z score: 5.4±2.4) had TSH>4mUI/l; fT3 levels were associated with age and transaminase activity; using multivariate regression analysis, with z-score and age as covariates, fT4 showed correlations with TSH, insulin, HOMA IR, blood lipids, and fibrinogen. No correlations were found with leptin, iodine excretion, IGF-I.In 79 patients followed for 52±15 wk (45% girls; age range 8-18.3 years), univariate regression showed a positive correlation between changes in TSH and HOMA-IR, and between changes in fT4 and HDL. Multivariate regression analysis with z score as covariate showed that baseline TSH was associated with negative changes in HOMA-IR. CONCLUSIONS: Increased TSH may be predictive of a decrease in insulin resistance, it should be measured with thyroid hormones; fT4 was associated with a low metabolic risk. Changes in thyroid function could protect against the occurrence of obesity-associated metabolic diseases.

Deferoxamine blocks death induced by glutathione depletion in PC 12 cells.

The purpose of the present work was to investigate the mechanisms by which glutathione depletion induced by treatment with buthionine sulfoximine (BSO) led within 24-30 h to PC 12 cells apoptosis. Our results showed that treatment by relatively low concentrations (10-30 µM) of deferoxamine (DFx), a natural iron-specific chelator, almost completely shielded the cells from BSO-induced toxicity and that DFx still remained protective when added up to 9-12h after BSO treatment. On the other hand, phosphopeptides derived from milk casein and known to carry iron across cell membranes, markedly potentiated the toxic action of BSO when loaded with iron but were ineffective in sodium form. Kept for 24 h in serum-free medium, the cells underwent a decrease in glutathione content after BSO treatment, but remained viable. However, these BSO-pre-treated cells showed a rapid (90-120 min) decrease in cell viability when incubated with low doses of iron, whereas a great proportion of them remained viable in the presence of higher concentrations of copper and zinc. We also observed in PC 12 cells an early (4-8 h) and transient increase in the expression of ferritin subunits following BSO addition. Taken together these results suggest that BSO-induced glutathione depletion leads to an alteration of cellular iron homeostasis, which may contribute to its toxicity.

Comparison of fat oxidation during exercise in lean and obese pubertal boys: clinical implications.

OBJECTIVE: To examine fat oxidation rates during exercise in lean and obese pubescent children. DESIGN: A graded leg cycle ergometry test was performed by two groups of pubescent boys (13 lean: mean (SD) age 12.0 (0.5) years, body mass index (BMI) 18.56 (1.12) kg/m(2); 17 obese: mean (SD) age 12.1 (0.1) years, BMI 26.68 (3.37) kg/m(2); p<0.001). The first step of the test was fixed at 30 W and power was gradually increased by 20 W every 3.5 min. The mean ventilatory gas measurement was obtained during the last 30 s of each step for calculation of fat oxidation rate vs exercise intensity. RESULTS: At low intensity (0-30% of peak oxygen consumption) when fat-free mass is considered, the fat oxidation rate was identical for the two groups. At higher intensities (40%, 50% and 60% of peak oxygen consumption) the fat oxidation rate was significantly higher in lean boys than in obese boys. CONCLUSION: These results confirm that obese pubertal boys have fat-free mass decreased capacities to use fat during moderate exercise. The findings suggest that obese boys need to practise physical activity at a lower intensity than healthy boys to enhance lipolysis and diminish adipose tissue and the consequences of obesity.

Interactions between ethylenediaminetetraacetic acid (EDTA) and iron absorption pathways, in the Caco-2 model.

UNLABELLED: EDTA is a well known enhancer of iron absorption; however, the precise way of absorption of iron ingested in presence of EDTA is not known; some data suggest it could use a passive, non regulated paracellular way. Iron (sulphate or gluconate) absorption by Caco-2 cells was assessed in presence of EDTA. EDTA did not change the apical uptake of iron; transport in the basal chamber increased by 98% for FeSO4 and 95% for Fe gluconate. By contrast, intracellular storage decreased by 31% for FeSO4 and 64% for Fe gluconate. In addition EDTA induced a significant increase of permeability of the cell monolayer assessed by a decrease of transepithelial electrical resistance: 314+/-34 Omegacm(-2) to 235+/-57 Omegacm(-2) for sulphate, 414+/-33 Omegacm(-2) to 223+/-36 Omegacm(-2) for gluconate; iron free control: 410+/-10 Omegacm(-2). CONCLUSIONS: These results suggest that in presence of EDTA iron absorption occurs mainly by the paracellular instead of the regulated cellular way, that could potentially enhance its toxicity.

Tracing of Enterobacter sakazakii isolates in infant milk formula processing by BOX-PCR genotyping.

AIMS: Enterobacter sakazakii is an emerging food-borne pathogen that can cause rare but severe neonatal meningitis, bacteraemia and necrotizing enterocolitis. Contaminated powder infant formulae (PIF) have been identified as one of various infection routes. In this study, E. sakazakii was monitored in the processing environment of a PIF factory to identify possible dissemination routes. METHODS AND RESULTS: The BOX-polymerase chain reaction (PCR), a fingerprinting technique which targets the repetitive BOX sequences, was used in routine to identify points of contamination and investigate clonal persistence. Two hundred E. sakazakii isolates were collected and typed. Most (70%) showed the same fingerprint that revealed the persistence of resident E. sakazakii strains in the processing environment. This method allowed to detect contamination of some PIF by dry-blending ingredients. CONCLUSIONS: Environment was the major cause for contamination of PIF and facilities. Some raw materials delivered as powder were also implicated. SIGNIFICANCE AND IMPACT OF THE STUDY: Routine BOX-PCR genotyping was very useful to trace and investigate in real-time dissemination of micro-organisms in the PIF plant and to implement a series of additional control measures to reduce the risk of final product contamination by E. sakazakii.

Genotypic characterization of Enterobacter sakazakii isolates by PFGE, BOX-PCR and sequencing of the fliC gene.

AIMS: Enterobacter sakazakii is an emerging food-borne pathogen that can cause rare but severe forms of neonatal meningitis, bacteraemia and necrotizing enterocolitis. A rapid typing method at the strain level is needed to determine the monoclonality or polyclonality of the isolates during outbreaks. METHODS AND RESULTS: The BOX-PCR fingerprinting technique, which targets the repetitive BOX sequences, and sequencing of the flagellin gene, fliC, were evaluated against a panel of 27 Ent. sakazakii strains from clinical and environmental sources. The typeability and discriminatory power of the techniques were compared with those of pulsed-field gel electrophoresis (PFGE), the reference genotyping method. BOX-PCR results yielded 92% agreement with PFGE results, whereas fliC gene sequencing was poorly discriminative. CONCLUSIONS: In our study, BOX-PCR and PFGE were similarly discriminatory to type Ent. sakazakii strains. The weak variability of the Ent. sakazakii fliC gene was related to the absence of the variable central domain present in most fliC genes of Enterobacteriaceae. SIGNIFICANCE AND IMPACT OF THE STUDY: The BOX-PCR typing provides an accurate discrimination and a rapid answer to identify clonal isolates of Ent. sakazakii.

[Treatment of pediatric obesity: effects on body composition and physical fitness]. / Prise en charge ambulatoire de l'obésité: effets sur la composition corporelle et la capacité aérobie.

AIMS: To assess the efficiency of an ambulatory weight management programme of pediatric obesity, including 1 gymnastic session per week, on body composition and physical fitness (max). SUBJECTS: Fifteen adolescents participated in the 9-month intervention. BMI and fitness and physical activity assessed by a questionnaire were evaluated at baseline, and after intervention. RESULTS: Prepubescent subjects (N=6): no significant change of BMI, body composition, nor max. Pubescent subjects: significant decrease of BMI, and z score BMI, and % fat mass, increase of fat free mass. Activity questionnaire: non-significant trend to decreased TV watching, significant increase in practice of physical activity during weekend. CONCLUSION: A modest increase in physical practice, included in the dietary-behavioural management of adolescent obesity, is able to improve overweight and physical fitness.

[High cord blood TSH in Morocco: iatrogenic hypothyroidism?]. / TSH au cordon élevées au Maroc: hypothyroïdie iatrogène?

A short prospective study was conducted to assess thyroid status in healthy full term newborns (n=90) of a large maternity of Marrakech (Morocco), as part of the validation of a national salt iodisation program. High TSH (>5mU/l) was detected in 89% of infants tested; urinary iodine excretion was measured in 35 of the mothers, and was found to be normal (100-200microg/l) or high (>200microg/l)(n=27) in all of them. Milk iodine concentration was measured in 315 lactating women from the same area. Low values (<41microg/l) were found in 60% of them. The common use of iodinated disinfectants during delivery could be responsible for the high urine iodine values of mothers; however iodine deficiency seems to remain a widespread problem in this population and justifies a large scale survey of iodine status.

Iron availability and consumption of tea, vervain and mint during weaning in Morocco.

BACKGROUND/AIMS: Iron deficiency impairs growth and psychomotor development of infants. In Morocco, infusions are introduced very early in infant diet, and could contribute to iron deficiency, due to their high polyphenol content. METHODS: The availability of tea, mint and vervain infusions was assessed using an in vitro model of digestion and dialysis. Two gastric pHs were used: pH 4 as in the first week life, and pH 2.5 as in older infants. Six repetitions of each experiment were made. The total polyphenol content of infusions was measured. RESULTS: At pH 4 and at pH 2.5, iron availability was decreased by tea and vervain, and increased by mint and ascorbic acid. At both pHs it was increased by addition of ascorbic acid to tea and vervain. In addition, at pH 2.5 it was increased by addition of ascorbic acid to mint. The highest value was observed in the presence of both ascorbic acid and mint (33.1 +/- 4.1%). In any case, iron availability was higher at pH 2.5 than at pH 4 (with single compounds or combinations with ascorbic acid). The polyphenol contents (mg/l) of tea, vervain and mint infusions were 2,236.1, 771.1, and 16.5. CONCLUSIONS: Tea and vervain infusions inhibited iron availability. In contrast, mint improved it; vitamin C helped in preventing these inhibiting properties. It could be proposed to discourage tea and vervain drinking at early weaning and to replace them by mint infusion, or at least to promote the consumption of vitamin C-rich fruit juice to counteract these inhibiting effects.

Influence of short-chain fatty acids on iron absorption by proximal colon.

BACKGROUND: Short-chain fatty acids produced by bacterial fermentation in the colon enhance the local absorption of cations, such as calcium, that could be used to improve the bioavailability of iron if a significant colonic absorption of iron were to occur. METHODS: Iron (iron gluconate, 100 microM) absorption by the caecum of the rat was compared with that in proximal sites of the small bowel using the Ussing chamber model; the influence of probiotic bacteria (Propionibacterium freudenreichii) on iron absorption was assessed and compared with that of two of their fermentation products (acetic and propionic acids) using the Ussing chamber and the ligated colon with gamma emitting iron as experimental models. RESULTS: The caecum absorbed less iron than the duodenum, but significantly more than the jejunum and ileum. This occurred mainly through an enhanced mucosal transfer of iron uptake. Propionibacteria enhanced iron absorption from the proximal colon; the same effect was observed in the presence of viable bacteria, or the culture medium free of viable bacteria, or acetate and propionate or propionate alone. CONCLUSIONS: The proximal colon could be a significant site available for iron absorption; this absorption can be enhanced by local production of short-chain fatty acids such as propionate.

Aluminium increases xanthine oxidase activity and disturbs antioxidant status in the rat.

The mechanisms responsible for the neurotoxic effects of Al remain poorly understood. In order to determine whether Al promotes oxidative stress in vivo, we measured the enzymatic activity of xanthine oxidase (XO), superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione-S-transferase (GST) and glutathione reductase (GR) in four groups of rats after eight days of intraperitoneal administration of variable concentrations of Al (0, 5, 10, and 15 mg/kg body weight, respectively). XO activity was measured in both plasma and liver samples, and the activities of the remaining enzymes were further determined in the brain and red blood cells (RBC). The most significant changes were observed in XO and GPX activities, that were enhanced and depressed, respectively. In both instances, the enzyme activities were correlated with Al concentrations, either positively (XO) or negatively (GPX). Enhancement of XO and inhibition of GPX activity may lead to the accumulation of intermediate toxic compounds such as hydrogen peroxide and hydroxyl radicals, since SOD activity is increased as well. The latter finding must be taken with some caution because previous studies have shown contradictory results in this field. Our data suggest that Al toxicity could be mediated by its action on both pro- and anti-oxidant enzymes. The biological significance of these findings remains to be established.

Inhibition of zinc absorption by iron depends on their ratio.

Previous studies upon zinc-iron interactions gave conflicting results that could come from differences in protocol design or in trace element status of subjects. The present work assessed the influence of zinc : iron ratio and iron deficiency upon zinc absorption. The digestive absorption of zinc sulphate (100 micromol Zn/l) in presence of iron gluconate was studied in perfused jejunal loops (n = 6/group) of normal rats (range 0-1000 micromol Fe/l) and iron deficient rats (200-750 micromol Fe/l). In normal rats no significant iron inhibition on zinc absorption occurred at Fe:Zn ratio below 2:1. At higher ratios zinc uptake and net absorption decreased significantly (p<0.05). Between 2:1 and 5:1 a dose dependent inhibition of zinc absorption occurred and reached a plateau beyond this ratio. In iron deficient animals no changes in zinc uptake, mucosal retention and absorption compared to normal animals occurred at ratio 2:1. At higher ratios differences were observed at every zinc absorption step except for mucosal retention at 7.5:1 ratio. Iron-zinc interactions depend on their ratio and on previous trace elements status of subjects. Due to the wide and unknown variations that were likely to occur between the subjects of previous human and experimental studies, these results could explain some of the discrepancies between their results.

Zinc and iron status and growth in healthy infants.

OBJECTIVE: To elucidate the relationships between growth and zinc and iron status in normal infants. STUDY DESIGN: Growth of normal infants (less than 3 y old: n = 66) was prospectively assessed with a mean delay of 24+/-6 weeks between measurements; subjects were free from illness and presented with a normal growth. Growth was compared to serum zinc (s-zinc), IGF-1 and iron status. SETTING: Teaching hospital of Caen. RESULTS: No relation was found between linear or weight growth and s-zinc; when taking into account the effect of age, linear growth was significantly associated with ferritin (P<0.001); weight gain was significantly correlated with IGF-1 (P = 0.034) and ferritin (P = 0.008). No relationship was found between s-zinc and iron status. CONCLUSIONS: In normal infants iron status, more than serum zinc, seems to be correlated with growth.

Influence of the extent of haemoglobin hydrolysis on the digestive absorption of haem iron in the rat. An in vitro study.

This study was designed to assess the haem-peptide interactions which occur during progressive haemoglobin hydrolysis by digestive enzymes and their relationship with haem iron digestive absorption. The behaviour of different haemoglobin hydrolysates was studied using the Ussing chamber model. Hydrolysates were produced from enzyme digestion of bovine haemoglobin at pH 3 by pepsin and at pH 10 by subtilisin. Samples with increasing degrees of hydrolysis (0-15 %) were studied. Biochemical assays (pyridine haemochromogen method and UV absorption spectra) were used to follow haem solubility and haem-peptide interactions in samples. Increasing the hydrolysis level of haemoglobin was associated with an enhanced iron uptake; the highest uptake rate was reached between 8 and 11 % of globin hydrolysis, whichever enzyme was used. The mechanisms rendering iron soluble and available differ between the two enzymes. The comparison between biochemical and absorption data suggests that the formation of soluble peptide-haem complexes was not sufficient to enhance haem iron absorption, since globin-bound iron is poorly absorbed; an efficient absorption occurred only when haem was loosely bound to low molecular weight peptides.

The effect of cysteine and 2,4-dinitrophenol on heme and nonheme absorption in a rat intestinal model.

Previous studies have showed that purified heme iron forms insoluble polymers that are poorly absorbed. The presence of peptides and of amino acids maintaining heme iron in a soluble form could improve its bioavailability. The digestive uptake and transfer of a concentrated hydrolysate of heme peptides (HPH) and of iron gluconate (Gluc) at 100 µM were compared in vitro in a Ussing chamber. The effects of an enhancing amino acid (L-cysteine) on the uptake and transfer of both forms were assessed. An inhibitor of the oxidative phosphorylation (2,4-dinitrophenol; DNP) was used to differentiate the active and passive mechanisms of the absorption. The mucosal uptake (%Tot) and enterocyte transfer (%S) of the two sources of iron did not differ. DNP significantly reduced %Tot and %S of both forms. Cysteine significantly enhanced %Tot and %S of HPH and Gluc, partly corrected the inhibition exerted by DNP on %Tot of HPH and %S of both forms, and fully restored %Tot of Gluc. In presence of peptides produced by globin hydrolysis, the absorption of hemoglobin iron was efficient; it was mostly energy dependent and, therefore, should have occurred by a regulated transcellular pathway. Cysteine enhanced the passive uptake of iron and the passive processes involved in the enterocyte transfer of the common pool made of both sources (heme and nonheme) of iron. These results showed that heme iron can be purified and concentrated without impairing its digestive absorption, provided it remains in presence of peptides or amino acids.

Iron tissue storage and hemoglobin levels of deficient rats repleted with iron bound to the caseinophosphopeptide 1-25 of beta-casein.

Caseinophosphopeptides (CPP) issued from enzyme digestion of caseins bind cations and keep them soluble in the digestive tract. They could be used as ligands to improve iron (Fe) bioavailability. Fe-deficient young rats were repleted with Fe (40 or 200 mg/kg of diet) bound either to the beta-CN (1-25) of beta-casein or to whole beta-casein or as FeSO(4). A control pair-fed group was given 200 mg of Fe (FeSO(4))/kg of diet for 6 weeks. After repletion, hemoglobin concentration of the control group was reached only by the ) animals fed 200 mg of Fe/kg; beta-CN (1-25) bound Fe (40 and 200 mg) produced higher Fe liver and spleen stores than FeSO(4). Binding Fe to the whole, nonhydrolyzed beta-casein gave results intermediate between the other experimental groups. Binding Fe to phosphoserine residues of low molecular weight CPP improved its ability to cure anemia and to restore iron tissue stores, as compared to Fe bound to the whole casein and to inorganic salts.

[Iron and zinc supplementation during pregnancy: interactions and requirements]. / Supplémentation en fer et zinc pendant la grossesse: interactions, besoins.

The metabolism of iron and zinc during pregnancy is reported; their deficiency impairs foetal growth, and increases the rate of complications at delivery and of preterm births. Providing iron and zinc supplements must take into account their metabolic interactions which are likely to decrease their bioavailability. Supplementations should be prescribed only during at risk pregnancy and their dosage should be adapted to lessen these side effects and improve their efficiency.

Early neurological and neuropsychological development of the preterm infant and polyunsaturated fatty acids supply.

OBJECTIVES: To compare the early neurological maturation of premature newborns (PT) fed breast milk (BM) or a formula containing only 18-carbon polyunsaturated fatty acids (PUFA) (A) or enriched with long chain (LC) PUFA (B). METHODS: PT enrolled the 2nd day of enteral feeding (D0) were fed BM (n = 15; 4 dropped out) or randomly assigned to A (n = 11; 2) or B (n = 14; 1) for at least 30 days (D30). Auditory and visual evoked potentials (EPs) and nerve conduction velocity (NCV) and plasma and red blood cell (RBC) phospholipid composition were determined at D0 and D30. No difference was found between groups for the D0-D30 changes in EP parameters. The maturation of motor NCV was slower in the B group than in the two other groups. In plasma, the changes were higher in B than in the BM and A groups for linoleic acid (P < 0.05), in BM versus B group for arachidonic acid (P < 0.02). In RBC, formula groups displayed higher linoleic acid level than the BM group (P < 0.05). No difference was found between groups for the changes in arachidonic and docosahexaenoic acids. CONCLUSIONS: A balanced supply of n-6 and n-3 PUFA without addition of LC-PUFA allowed an adequate early maturation of the central nerve system. The effects of LC-PUFA on the maturation of NCV remain to be confirmed.