Ulrike Albers

Sampling and processing of fresh blood samples within a European multicenter nutritional study: Evaluation of biomarker stability during transport and storage

Introduction:Analysis of several biological markers improves the quality and physiologic comprehension of data obtained in epidemiological nutritional studies.Aim:To develop a methodology that guarantees the centralized analysis and quality assurance of the most relevant blood parameters from fresh blood samples in adolescents in a European multicenter study.Materials and methods:Stability of selected nutrients and biomarkers (vitamins, fatty acids, iron metabolism and immunological parameters) chosen with respect to time and temperature of sample transport and storage was evaluated as part of the pilot study of the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) project.Results:Routine biochemistry and iron status parameters included in the HELENA Cross-Sectional Study (CSS) protocol could be analyzed within 24 h from fresh blood samples without any stability problems (coefficient of variation (CV)<5%, P<0.05). However, stability tests for lymphocyte subpopulations, vitamin C and fatty acids showed that they are very unstable at room temperature without any treatment. Therefore, a special handling for these samples was developed. Vitamin C was stabilized with metaphosphoric acid and transported under cooled conditions (CV 4.4%, recovery rate >93%, P>0.05). According to the results, a specific methodology and transport system were developed to collect blood samples at schools in 10 European cities and to send them to the centralized laboratory (IEL, Bonn, Germany). To guarantee good clinical practice, the field workers were instructed in a training workshop and a manual of operation was developed.Conclusion:The handling and transport system for fresh blood samples developed for the European multicenter study HELENA is adequate for the final part of the HELENA-CSS and will provide, for the first time, reference values for several biological markers in European adolescents.

Prevalence of overweight and obesity in non-institutionalized people aged 65 or over from Spain: the elderly EXERNET multi-centre study

Coupled with the growth of the older population, an increase in the prevalence of overweight and obesity in this age group has occurred in the last decades. The main aims of the present study were (i) to provide an updated prevalence of overweight and obesity in a representative sample of the Spanish elderly population; (ii) to calculate the prevalence of sarcopenic obesity (SO) and (iii) to analyse the relationships between adiposity measurements and lifestyle. A cross‐sectional study was carried out in a sample of 3136 persons representative of the non‐institutionalized population ≥65 years of age. Anthropometric measurements were obtained using standardized techniques and equipment. Overall, 84% of the population can be categorized as overweight and/or obese. The present study indicates that 67% of the Spanish elderly population has an increased percentage of fat mass and more than 56% suffer from central obesity. Moreover, SO is present in 15% of the Spanish elderly population. Finally, a strong relationship between both physically active and sedentary lifestyles and the level of adiposity was found. Prevalence of overweight and obesity among elderly people in Spain is very high and is still increasing. Lifestyle seems to be a determinant factor in the development of obesity among elderly people.

B-Vitamins and Homocysteine in Spanish Institutionalized Elderly

BACKGROUND Hyperhomocysteinemia is an accepted risk factor for cardiovascular disease, and possibly also for cognitive impairment and dementia. It has also been proposed as a marker for the status of the B vitamins, which participate in the metabolism of homocysteine. Therefore, especially in the elderly, it is important to know the prevalence of high homocysteine (tHcy) levels and the influence that B vitamins have on them. MATERIAL AND METHODS 218 elderly of both sexes, aged 60-105, living in an elderly home in Granada (Spain), were screened for serum folate, red blood cell (RBC) folate, serum cobalamin (B12) (Abbott, IMx), holotranscobalamin II (Holo-TC II) (HoloTC RIA, Axis-Shield), methylmalonic acid (MMA) (MS-GC), total pyridoxine (B6) (HPLC), and total homocysteine (tHcy) (Abbott, IMx). RESULTS Hyperhomocysteinemia (tHcy >12 pmol/L) was detected in 80.7%. Serum folate deficiency was severe (< or =4 ng/mL) in 19.3% and moderate (4-7 ng/mL) in 43.1%. In 14.2% of the elderly RBC folate was < or =175 ng/mL, and in 61.0% it was between 175-400 ng/mL. Vitamin B12, measured in serum (< or =200 pg/mL), was deficient in 15.8%, but if measured as Holo-TC II (< or =45 pmol/L), deficiency ranged up to 39.1%. MMA was high (> or =300 nmol/L) in 45.6%. Vitamin B6 (< 20 nmol/L) was low only in one person. In order to identify the factors that could predict tHcy levels, a multiple regression analysis was performed. Best results corresponded to the combination of log serum folate and log Holo-TC II, which gave values of R > 0.5. If analyzed independently, the highest correlation was with log serum folate (r = -0.290), followed by RBC folate (r = -0.263), Holo-TC II (r = -0.228), log B12 (r = -0.175), and log B6 (r = -0.078). CONCLUSION There is a high prevalence of vitamin B deficiency and hyperhomocysteinemia in the studied population. Our data confirm the influence of these vitamins, especially folate, on tHcy levels, but hyperhomocysteinemia cannot be used as the only diagnostic criterion to detect subclinical vitamin deficiency in elderly people, especially to detect vitamin B12 deficiency.

Gender and age influence blood folate, vitamin B12, vitamin B6, and homocysteine levels in European adolescents: the Helena Study.

It is important to be able to evaluate vitamin status correctly at any age, but this is especially vital during adolescence since there are higher requirements for healthy growth and development. However, there are no currently available B-vitamin reference values for healthy adolescents. The aim of the present study is to assess the vitamin B status in European adolescents in order to contribute to the development of reference values for selected B-vitamins and total homocysteine (tHcy). Within the HELENA (Healthy Lifestyle in Europe by Nutrition in Adolescence) cross-sectional study, a sub sample of 1051 (499 males, 552 females) adolescents from ten European cities aged 12.5 to 17.49 were analyzed for fasting plasma folate (PF), red blood cell (RBC) folate, serum cobalamin (Cbl), holo-transcobalamin (Holo-TC), Vitamin B(6) (PLP), and tHcy. The level of significance was set at P < .05. Following the current cut-off for adults, 2% had low Cbl and 5% had low holo-TC concentrations. Low concentrations of both PF and RBC folate were identified in 10%. Five percent had PLP concentrations <20 nmol/L and 20% <30 nmol/L. Moreover, 5% had high tHcy; median values for the whole sample were: PF 16.0 nmol/L, RBC folate 721.9 nmol/L, Cbl 319 pmol/L, Holo-TC 57.8 pmol/L, and tHcy 6.7 μmol/L. Females had significantly higher median Cbl but lower PLP and tHcy concentrations (P < .01). THcy increased (P < .001) and PF (P < .001) concentrations decreased across age categories. Subjects showed significantly higher tHcy values at the fifth percentile of PF, corresponding with 7.5 nmol/L. Sex and age had an influence on most of the studied biomarkers and should be taken into account. The HELENA percentile distribution is consistent with data from smaller studies and could be used as reference value to characterize B-vitamin status of European adolescents.

Reference values for leptin, cortisol, insulin and glucose, among European adolescents and their association with adiposity: The HELENA Study

BACKGROUND AND OBJECTIVE Adequate concentrations of leptin, cortisol, and insulin are important for a suitable metabolism and development during adolescence. These hormones jointly with glucose play a major role in fat metabolism and development of childhood obesity. Our main objective was to quantify biomarkers as leptin, cortisol, insulin and glucose status in European adolescents to contribute to establish reference ranges. METHODS A representative sample of 927 adolescents (45% males, 14.9±1.2 years for the overall population) from ten European cities of the HELENA study was used to obtain fasting blood samples for these biomarkers. The percentile distributions were computed by sex and age and percentiles were associated with BMI classification. RESULTS Serum leptin concentration in adolescents varied significantly according to BMI, sex and age (all p < 0.001). Cortisol presented a tendency to increase with age, both for females and males, while insulin and glucose were stable with age. Leptin and insulin were highest in obese adolescents (p < 0.001), whilst cortisol and glucose did not vary with BMI. Percentiles 5, 25, 50, 75 and 95, for hormones values were, respectively: 1.27, 4.06, 11.54, 26.70 and 65.33 ng/ml for leptin; 5.00, 8.11, 11.14, 15.00 and 24.51 μg/dl for cortisol and 3.65, 6.15, 8.52, 11.90 and 20.53 μlU/ml for insulin. CONCLUSIONS In adolescents, leptin, cortisol, insulin and glucose concentrations are differently affected by age, sex and BMI. Establishment of reference ranges (percentiles) of these biomarkers would be of great interest when pediatricians have to assess the trend of an adolescent to develop obesity years after.

Harmonization process and reliability assessment of anthropometric measurements in the elderly EXERNET multi-centre study.

Background The elderly EXERNET multi-centre study aims to collect normative anthropometric data for old functionally independent adults living in Spain. Purpose To describe the standardization process and reliability of the anthropometric measurements carried out in the pilot study and during the final workshop, examining both intra- and inter-rater errors for measurements. Materials and Methods A total of 98 elderly from five different regions participated in the intra-rater error assessment, and 10 different seniors living in the city of Toledo (Spain) participated in the inter-rater assessment. We examined both intra- and inter-rater errors for heights and circumferences. Results For height, intra-rater technical errors of measurement (TEMs) were smaller than 0.25 cm. For circumferences and knee height, TEMs were smaller than 1 cm, except for waist circumference in the city of Cáceres. Reliability for heights and circumferences was greater than 98% in all cases. Inter-rater TEMs were 0.61 cm for height, 0.75 cm for knee-height and ranged between 2.70 and 3.09 cm for the circumferences measured. Inter-rater reliabilities for anthropometric measurements were always higher than 90%. Conclusion The harmonization process, including the workshop and pilot study, guarantee the quality of the anthropometric measurements in the elderly EXERNET multi-centre study. High reliability and low TEM may be expected when assessing anthropometry in elderly population.

EL EJERCICIO AGUDO AUMENTA LAS CONCENTRACIONES DE HOMOCISTEÍNA EN VARONES FÍSICAMENTE ACTIVOS

INTRODUCTION High levels of homocysteine (Hcy) have been identified as a cardiovascular risk factor. Regarding physical exercise, the results are contradictory. OBJECTIVES The aim of this study was to determine the influence of maximal intensity exercise and submaximal constant exercise on total serum homocysteine concentrations (tHcy) and other related parameters. MATERIAL AND METHODS Ten physically active male subjects (mean age: 23.51 ± 1.84), performed two treadmill tests, a maximal test and a stable submaximal test at an intensity of 65% of maximal oxygen uptake (VO2max). Serum concentrations of tHcy, Folate, Vitamin B12 and creatinine were analysed before and after each test. RESULTS Significant increase in serum tHcy concentrations after the maximal (p < 0.05) and submaximal (p < 0.01) tests were observed. Folate and vitamin B12 concentrations also increased significantly after both tests (p < 0.05). Creatinine levels increased only after the maximal test (p < 0.001). A statistically significant inverse relationship was found between folate and tHcy concentrations (p < 0.05) at all the measurement points. CONCLUSION THcy levels increased significantly after acute exercise in both maximum and submaximal intensity exercises.

Physical strength is associated with Mini-Mental State Examination scores in Spanish institutionalized elderly.

The present cross‐sectional study aimed at assessing muscle strength of hands, the dominant arm and legs in Spanish institutionalized elderly people according to sex, age and cognitive status.

Do MCI patients with vitamin B12 deficiency have distinctive cognitive deficits

BackgroundVitamin B12 deficiency is common in older people, and may be responsible for reversible dementia. Low serum vitamin B12 levels were also observed in patients with Mild Cognitive Impairment (MCI). It is not known whether patients with vitamin B12 deficiency have a distinctive profile of cognitive impairment different from the episodic memory deficit usually observed in MCI.ResultsFrom a cohort of 310 patients with MCI followed in a memory clinic in Lisbon, only 10 cases with vitamin B12 deficiency were found. From collaboration with other neurologists, 5 further patients with vitamin B12 deficiency were added. These cases were compared to MCI patients with normal vitamin B12 levels in a ratio 1:3. The duration of subjective cognitive symptoms was significantly shorter in MCI patients with B12 deficiency (1.2±1.0 years) as compared to MCI patients with normal vitamin B12 levels (3.4±3.0 years, p<0.001, Student’ t test). There were no statistically significant differences in the neuropsychological tests between MCI patients with and without vitamin B12 deficiency. Vitamin B12 was started in MCI patients with vitamin B12 deficiency, with no noticeable clinical improvement.ConclusionMCI patients with low levels of vitamin B12 had no particular profile of cognitive impairment, however vitamin B12 deficiency might have precipitated the onset of symptoms. The effect of vitamin B12 supplementation in patients with MCI and low vitamin B12 levels should be clarified by future prospective studies.

Efficacy of a 28-day oral cyanocobalamin supplementation on vitamin B status in Spanish institutionalized elderly.

BACKGROUND Cobalamin deficiency is a common problem in the elderly. There is no consensus about adequate doses for supplementation. SUBJECTS/METHODS We performed an intervention study in order to establish the efficacy of a supplement providing 500 µg cyanocobalamin for four weeks in sixty-four institutionalized elderly residents, over 60 years of age, in Madrid (Spain). Before and after treatment, concentrations of serum cobalamin, serum holotranscobalamin, serum total homocysteine, and serum and red blood cell folate were analyzed. Clusters were built according to the initial cobalamin status and differences in the effect of supplementation were checked using a general linear model for repeated measures. RESULTS Cobalamin and holotranscobalamin increased highly significantly from 308 to 558 pmol/L and from 54 to 96 pmol/L (p < 0.001) in the whole study group as well as in each subgroup (clustered by initial cobalamin levels, all p < 0.01), with the highest relative change in the subgroup with the lowest initial cobalamin values. Total homocysteine decreased from 15 to 13 µmol/l, p < 0.001). Only the change of cobalamin (F = 4.61, p < 0.01), but not of holotranscobalamin nor total homocysteine, depended on the initial serum cobalamin status. CONCLUSIONS A supplementation with an oral supplement solution of 500 µg cyanocobalamin daily for only four weeks, a shorter period than that found in former studies, may be considered suitable in institutionalized elderly.