Including non-dietary sources into an exposure assessment of the European Food Safety Authority: The challenge of multi-sector chemicals such as Bisphenol A.

In the most recent risk assessment for Bisphenol A for the first time a multi-route aggregate exposure assessment was conducted by the European Food Safety Authority. This assessment includes exposure via dietary sources, and also contributions of the most important non-dietary sources. Both average and high aggregate exposure were calculated by source-to-dose modeling (forward calculation) for different age groups and compared with estimates based on urinary biomonitoring data (backward calculation). The aggregate exposure estimates obtained by forward and backward modeling are in the same order of magnitude, with forward modeling yielding higher estimates associated with larger uncertainty. Yet, only forward modeling can indicate the relative contribution of different sources. Dietary exposure, especially via canned food, appears to be the most important exposure source and, based on the central aggregate exposure estimates, contributes around 90% to internal exposure to total (conjugated plus unconjugated) BPA. Dermal exposure via thermal paper and to a lesser extent via cosmetic products may contribute around 10% for some age groups. The uncertainty around these estimates is considerable, but since after dermal absorption a first-pass metabolism of BPA by conjugation is lacking, dermal sources may be of equal or even higher toxicological relevance than dietary sources.

Association of heart rate and blood pressure among European adolescents with usual food consumption: The HELENA study.

BACKGROUND AND AIM: In adults, there is some evidence that improving diet reduces blood pressure (BP) and the subsequent risk of cardiovascular diseases (CVDs). However, studies that analyse this association in adolescents are still scarce. The objective of the present study was to examine the associations between heart rate, systolic (SBP), diastolic (DBP) and mean arterial blood pressure (MAP) among European adolescents and usual intake of vegetables, fruits, dairy products, meat, fish, high-sugar foods and savoury snacks. METHODS AND RESULTS: In total, 2283 adolescents from the HELENA-study (12.5-17.5 years old; 1253 girls) were included. Dietary intake was assessed using two computerized 24-hour dietary recalls. Age, sex, body mass index, maternal educational level, physical activity and Tanner stage were considered as confounders. Associations were examined by mixed model analysis stratified by sex. Tests for trend were assessed by tertiles of intake while controlling for the aforementioned confounders. Dairy products and fish intake were negatively associated with BP and heart rate. Significant decreasing trends were observed for heart rate and BP across tertiles of dairy products, fish intake and high-sugar foods intake (p < 0.05). Significant increasing trends were observed for SBP and MAP across tertiles of savoury snack intake (p < 0.05). CONCLUSION: Significant but small inverse associations between fish and dairy products consumption with blood pressure and heart rate have been found in European adolescents. Dietary intervention studies are needed to explore these associations in the context of the modification of several risk factors for the prevention of cardiovascular diseases.

Predictors of newly diagnosed atrial fibrillation in cryptogenic stroke: a cohort study.

BACKGROUND AND PURPOSE: A significant proportion of cryptogenic ischaemic strokes are due to paroxysmal atrial fibrillation (AF). As paroxysmal AF appears to inexorably progress to persistent or permanent AF, this study with long-term follow-up was designed to establish the profile of patients who developed AF after hospital discharge. METHODS: All patients with cryptogenic ischaemic stroke over a 1-year period were included (n = 164). Patients were prospectively followed up at the outpatient clinic. Information on long-term outcome included the presence of newly diagnosed AF (NDAF). A specific NDAF assessment was performed at least 2 years after the index stroke using a structured telephone interview. Baseline clinical, laboratory, and echocardiographic data of these patients were retrospectively recorded. Independent predictive factors were then used to produce a predictive grading score for NDAF, derived by logistic regression analysis. RESULTS: With a median follow-up of 854 days, 22 cases of NDAF (13%) were observed. On multivariate analysis, factors associated with NDAF were age ≥72 years (two points), history of coronary artery disease (one point) or stroke (one point), and left atrial area ≥16 cm(2) (two points) (total score ranging from 0 to 6). Patients with a score ≤1 point did not have NDAF during follow-up. CONCLUSIONS: In cryptogenic ischaemic stroke, the NDAF score can be used to target patients at high risk of developing AF after hospital discharge, as a score of 0-1 was highly predictive of the absence of NDAF during follow-up. These results need to be confirmed in prospective studies.

Dupuytren's disease presentation, referral pathways and resource utilisation in Europe: regional analysis of a surgeon survey and patient chart review.

AIM: We explored the management of Dupuytren's disease (DD) using a surgeon survey and patient chart review. METHODS: Twelve countries participated: Denmark, Finland, Sweden (Nordic region); Czech Republic, Hungary, Poland (East); France, Germany, the Netherlands, United Kingdom (West); Italy, Spain (Mediterranean). A random sample of orthopaedic/plastic surgeons (N = 687) with 3-30 years' experience was asked about Dupuytren's contracture procedures performed during the previous 12 months. Information ≤ 5 consecutive patients per surgeon was extracted from patient charts (N = 3357). RESULTS: Overall, 84% of participants were orthopaedic surgeons; 56% of surgeons were hand specialists. Deciding factors for fasciectomy and dermofasciectomy were consistent across regions: metacarpophalangeal (MP) or proximal interphalangeal contracture > 45°, recurrent contracture, and high expectations for success. Deciding factors for percutaneous needle fasciotomy were less consistent across regions, but the leading factor was MP flexion < 20°. Overall, 49% of diagnoses and 55% of referrals were made by a general practitioner (GP), with regional variation: 31-77% for GP diagnoses and 36-81% for GP referrals. There were also differences in admission status (e.g. 9% of Nordic patients and 80% of Eastern patients were treated as inpatients). Most patients were treated in public hospitals and most procedures were covered by public health insurance. CONCLUSIONS: We found regional variations in surgical practice, patient characteristics and referral patterns. Understanding current diagnosis and treatment patterns, in relation to regional differences in health economics, may improve physicians' diagnosis of DD and guide patients towards appropriate, customised management plans.

Surgical management of Dupuytren's contracture in Europe: regional analysis of a surgeon survey and patient chart review.

AIM: We explored regional variations in the surgical management of patients with Dupuytren's contracture (DC) in 12 European countries using a surgeon survey and patient chart review. METHODS: Twelve countries participated: Denmark, Finland, Sweden (Nordic region); Czech Republic, Hungary, Poland (East); France, Germany, the Netherlands, UK (West); Italy, Spain (Mediterranean). For the survey, a random sample of orthopaedic/plastic surgeons (n = 687) with 3-30 years' experience was asked about DC procedures performed during the previous 12 months. For the chart review (n = 3357), information from up to five consecutive patients was extracted. Descriptive statistics are reported. RESULTS: Ninety-five per cent of all surgeons used fasciectomy for DC, followed by fasciotomy (70%), dermofasciectomy (38%) and percutaneous needle fasciotomy (35%). Most surgeons were satisfied with fasciectomy over other procedures. Recommended time away from work and duration of physical therapy increased with the invasiveness of the procedure. The intra-operative complication rate was 4.0%; the postoperative complication rate was 34%. Overall, ≥ 97% of the procedures were rated by surgeons as having a positive outcome. Across all regions, 54% of patients had no nodules or contracture after the procedures. Only 2% of patients required retreatment within the first year of surgery. Important inter- and intraregional differences in these aspects of patient management are described. CONCLUSIONS: Understanding current regional treatment patterns and their relationships to country-specific health systems may facilitate earlier identification of, and intervention for, DD and help to optimise the overall treatment for patients with this chronic condition.

Neuropsychological assessment and cerebral vascular disease: the new standards.

Vascular cognitive impairment (VCI) includes vascular dementia (VaD), vascular mild cognitive impairment (VaMCI) and mixed dementia. In clinical practice, VCI concerns patients referred for clinical stroke or cognitive complaint. To improve the characterization of VCI and to refine its diagnostic criteria, an international group has elaborated a new standardized evaluation battery of clinical, cognitive, behavioral and neuroradiological data which now constitutes the reference battery. The adaption of the battery for French-speaking subjects is reported as well as preliminary results of the on-going validation study of the GRECOG-VASC group [Clinical Trial NCT01339195]. The diagnostic accuracy of various screening tests is reviewed and showed an overall sub-optimal sensitivity (<0.8). Thus, the general recommendation is to perform systematically a comprehensive assessment in stroke patients at risk of VCI. Furthermore,the use of a structured interview has been shown to increase the detection of dementia. In addition to the well known NINDS-AIREN criteria of VaD, criteria of VCI have been recently proposed which are based on the demonstration of a cognitive disorder by neuropsychological testing and either history of clinical stroke or presence of vascular lesion by neuroimaging suggestive of a link between cognitive impairment and vascular disease. A memory deficit is no longer required for the diagnosis of VaD as it is based on the cognitive decline concerning two or more domains that affect activities of daily living. Both VaMCI and VaD are classified as probable or possible. These new criteria have yet to be validated. Considerable uncertainties remain regarding the determinant of VCI, and especially the lesion amount inducing VCI and VaD. The interaction between lesion amount and its location is currently re-examined using recent techniques for the analysis of MRI data. The high frequency of associated Alzheimer pathology is now assessable in vivo using amyloid imaging. The first studies showed that about a third of patients with VaD due to small vessel disease or with poststroke dementia have amyloid PET imaging suggestive of AD. These new techniques will examine the interaction between vascular lesions and promotion of amyloid deposition. Although results of these on-going studies will be available in few years, these data indicate that efforts should be done in clinical practice to reduce underdiagnosis of VCI; VCI should be examined using a specific protocol which will be fully normalized soon for French-speaking patients; the sub-optimal sensitivity of screening tests prompts to use a structured interview to grade Rankin scale and to perform systematically a comprehensive assessment in stroke patients at risk of VCI; poststroke dementia occurring after 3 months poststroke may be preventable by treatment of modifiable vascular risk factors and secondary prevention of stroke recurrence according to recent recommendations.

[Androgen deprivation and cardiovascular risk in prostate cancer treatment]. / Hormonothérapie et risque cardiaque dans le traitement des cancers prostatiques.

Androgen suppression clearly increases the occurrence of cardiovascular risk factors : increased body fat, dyslipidemia and type II diabetes. Thus, several studies (but not all), showed an increase in coronary artery disease but also of sudden death and ventricular arrhythmias in relation to androgen deprivation, even for a short duration. This risk is particularly important in patients with existing cardiovascular risk factors or a history of heart disease. Cardiovascular risk should be balanced with the benefit of androgen deprivation on overall survival, especially when it is proposed in adjuvant setting, combined with radiotherapy in locally advanced prostate tumors. In practice, it is recommended that patients be referred to their physician for an evaluation before starting treatment, then 3 to 6 months after starting treatment, then once a year. The initial assessment should include: a clinical examination (with measurement of blood pressure and body index) and laboratory test with full lipid profile (total cholesterol, HDL and LDL cholesterol, triglycerides) and glucose. It is also important that patients with heart disease, receive lifestyle advice and low- dose aspirin (80 mg/day).

Surgical restoration of the upper limb in cervical spinal cord injury patients.

Prior to the 1950s, relatively few patients who suffered a transection of the cervical spinal cord survived their injury. Improved medical care and better coordination have resulted in greater numbers of patients surviving and leaving the hospital. The pioneering work of individual surgeons during the 1960s and 1970s stimulated interest in surgical restoration of upper limb function in tetraplegic patients. Since the publication of Moberg's monograph in 1978, surgical improvement of the upper limbs is regarded as one of the options that should be offered to tetraplegic individuals to improve their function. Patients are classified according to the level of spinal cord injury and the residual motor function (international classification: groups 1-9). Surgical procedures are adapted to the motor level for each group of patients. Indications for these procedures are well standardized, the techniques are well mastered, and predictable results can be expected. New nerve transfer techniques have been developed in recent years; they are currently being evaluated.

Early vs late surgical treatment of radial instability of the thumb metacarpophalangeal joint. A retrospective cohort study.

There is no clear evidence in the literature whether treating thumb radial collateral ligament (RCL) injury in the acute phase improves outcome. The purpose of the present study was to compare the clinical and radiological results of RCL repair in acute and chronic settings. Fourteen patients were included. Clinical range of motion (ROM) of the thumb, Kapandji score and radiological parameters were compared pre- and post-operatively to the contralateral uninjured thumb (control group) to evaluate the results of the surgical technique. Patients were then divided into two groups according to early versus late repair and outcomes were compared between the two groups and the control group. Preoperatively, mean spontaneous angle between first metacarpal (M1) and proximal phalanx (P1) (spontaneous M1P1 angle), ulnar stress M1P1 angle, ROM and Kapandji score differed significantly between injured and uninjured sides. Postoperatively these parameters for the injured side improved, reaching values similar to those on the uninjured side, especially with acute phase treatment; late treatment also tended to provide clinical improvement in ROM, Kapandji score and ulnar stress angle, but with significant improvement only for spontaneous deviation of the thumb. This study showed the late and immediate repair of the RCL of the thumb both gave good results, with slightly better outcome with acute phase repair. LEVEL OF EVIDENCE: Therapeutic, Level III.

Prognostic utility of the assessment of diastolic function in patients undergoing cardiac resynchronization therapy.

Conflicting data exist about the relationship between cardiac resynchronization therapy (CRT) and diastolic function. Aims of the study are to assess diastolic patterns in patients undergoing CRT according to the 2016 recommendations of the American Society of Echocardiography/European Association of Cardiovascular Imaging and to evaluate the prognostic value of diastolic dysfunction (DD) in CRT candidates. METHODS AND RESULTS: One-hundred ninety-three patients (age: 67 ± 11 years, QRS width: 167 ± 21 ms) were included in this multicentre prospective study. Mitral filling pattern, mitral tissue Doppler velocity, tricuspid regurgitation velocity, and indexed left atrial volume were used to classify DD from grade I to III. CRT-response, defined as a reduction of left ventricular (LV) end-systolic volume > 15% at 6-month follow-up (FU), occurred in 132 (68%) patients. The primary endpoint was a composite of heart transplantation, LV assisted device implantation, or all-cause death during FU and occurred in 29 (15%) patients. CRT was associated with a degradation of DD in non-responders. At multivariable analysis corrected for clinical variables, QRS duration, mitral regurgitation, CRT-response and LV dyssynchrony, grade I DD was associated with a better outcome (HR 0.37, 95% CI: 0.14-0.96). Non-responders with grade II-III DD had the worse prognosis (HR 4.36, 95%CI: 2.10-9.06). CONCLUSIONS: The evaluation of DD in CRT candidates allows the prognostic stratification of patients, independently from CRT-response.

Incidence of paroxysmal atrial tachycardias in patients treated with cardiac resynchronization therapy and continuously monitored by device diagnostics.

AIMS: Little is known about the incidence of paroxysmal atrial tachycardias (PAT) in patients with heart failure (HF). The availability of cardiac resynchronization therapy (CRT) devices with extended diagnostics for AT enables continuous monitoring of PAT episodes. The aim of the study was to assess the incidence over time of PAT in HF patients treated with CRT. METHODS AND RESULTS: Consecutive patients in NYHA functional class III or IV despite optimal drug therapy, QRS duration > or = 130 ms, left ventricular ejection fraction < or = 35%, and left ventricular end-diastolic dimension > or = 55 mm were eligible for enrolment. Patients with permanent or persistent atrial fibrillation (AF) were not included in the study. The first follow-up examination was performed 2 weeks after implantation, to optimize atrial sensing and CRT. Subsequent follow-up examinations were carried out 15 and 28 weeks after implantation, to collect the telemetric data. A total of 173 patients (67 +/- 11 years, M 116) were enrolled. Complete arrhythmia monitoring data were available from 120 patients over a mean follow-up of 183 +/- 23 days. Atrial tachycardia episodes were detected through telemetry in 25 of 120 patients (21%) during at least one follow-up examination. Atrial tachycardia episodes were recorded in 29 and 17% (P = NS) of patients with and without previous history of AF, respectively. CONCLUSION: More than 20% of the overall HF patient population treated with CRT suffer PAT episodes. Paroxysmal atrial tachycardia may interfere with response to CRT. Therefore, telemetric data may be relevant to drive the appropriate therapy in each patient.

The effects of improving low dietary protein utilization on the proteome of lamb tissues.

Cistus ladanifer L. is a common shrub endemic to the Mediterranean region with high levels of condensed tannins (CT). CT form complexes with dietary protein resisting microbial degradation in the rumen, which enhances dietary protein utilization in ruminant diets. The objective of this study was to evaluate the utilization of CT in the diet of lambs on the proteomes of muscle, hepatic and adipose tissues. Twenty-four Merino Branco ram lambs were randomly allocated to three treatments (n = 8): C - control (160 g crude protein (CP)) per kg DM, RP - reduced protein (120 g CP/kg DM); and RPCT - reduced protein (120 g CP/kg DM) treated with CT extract. At the end of the trial, lambs were slaughtered and the longissimus lumborum muscle, hepatic and peri-renal adipose tissues sampled. A two-way approach was used for proteomic analysis: 2D-DIGE and nanoLC-MS. In the muscle, C lambs had lower abundance proteins that partake in the glycolysis pathway than the lambs of other treatments. Control lambs had lower abundance of Fe-carrying proteins in the hepatic tissue than RP and RPCT lambs. The latter lambs had highest abundance of hepatic flavin reductase. In the adipose tissue, C lambs had lowest abundance of fatty-acid synthase. SIGNIFICANCE: soybean meal is an expensive feedstuff in which intensive animal production systems heavily rely on. It is a source of protein extensively degraded in the rumen, leading to efficiency losses on dietary protein utilization during digestion. Protection of dietary protein from extensive ruminal degradation throughout the use of plants or extracts rich in CT allow an increase in the digestive utilization of feed proteins. In addition to enhance the protein digestive utilization, dietary CT may induce other beneficial effects in ruminants such as the improvement of the antioxidant status.

FAO/WHO GIFT (Global Individual Food consumption data Tool): a global repository for harmonised individual quantitative food consumption studies.

Knowing who eats what, understanding the various eating habits of different population groups, according to the geographical area, is critical to develop evidence-based policies for nutrition and food safety. The FAO/WHO Global Individual Food consumption data Tool (FAO/WHO GIFT) is a novel open-access online platform, hosted by FAO and supported by WHO, providing access to harmonised individual quantitative food consumption (IQFC) data, especially in low- and middle-income countries (LMIC). FAO/WHO GIFT is a growing repository, which will serve as the global FAO/WHO hub to disseminate IQFC microdata. Currently five datasets from LMIC are available for dissemination, and an additional fifty datasets will be made available by 2022. To facilitate the use of these data by policy makers, ready-to-use food-based indicators are provided for an overview of key data according to population segments and food groups. FAO/WHO GIFT also provides an inventory of existing IQFC data worldwide, which currently contains detailed information on 188 surveys conducted in seventy-two countries. In order for end-users to be able to aggregate the available data, all datasets are harmonised with the European Food Safety Authority's food classification and description system FoodEx2 (modified for global use). This harmonisation is aimed at enhancing the consistency and reliability of nutrient intake and dietary exposure assessments. FAO/WHO GIFT is developed in synergy with other global initiatives aimed at increasing the quality, availability and use of IQFC data in LMIC to enable evidence-based decision-making and policy development for better nutrition and food safety.

The muscular, hepatic and adipose tissues proteomes in muskox (Ovibos moschatus): Differences between males and females.

The muskox (Ovibos moschatus) is a ruminant highly adapted to arctic conditions. The objective of this work is to study liver, muscle and adipose tissues proteomes in muskoxen highlighting sex differences. Ten animals (5 per sex) were sampled in Western Greenland during the winter hunting season. During carcass processing, muscle, liver and rump fat samples were obtained. Proteomic analyses were conducted using both gel-based and gel-free approaches. Gel-free data are available (ProteomeXchange; PXD014147). For gel-free analysis, 729, 853 and 792 proteins were identified for fat, liver and muscle, respectively. Several proteins were detected with differential abundance between male and female tissues: 77, 15 and 12 proteins using gel-free for adipose tissue, liver and muscle respectively while 3 differential proteins were identified in the gel-based analysis of the adipose tissue. Females have higher abundance of proteins involved in tissue structural stability in the muscle, while males have higher abundance of proteins related to muscle development. In the liver and adipose tissue, females have higher abundance of proteins related to oxidative-stress resistance. Proteins accumulated in the adipose tissue of males highlight higher adipogenic potential. Sex dimorphism is inherent to this species, with higher abundance of proteins in specific metabolic pathways. SIGNIFICANCE: The proteomes of the muskox muscle, hepatic and adipose tissues are characterized for the first time. In addition, the effect of sex on tissue protein abundance is studied. Our results reveal that sex dimorphism goes from morphology to the molecular level in this species, affecting protein, lipid and carbohydrate metabolism. This contributes for an in-depth look into sex dimorphism using proteomics which is lacking in most mammals, apart from model species. Moreover, this information has been related to nutritional status, which is particularly important when managing the muskox population and the transformation of its habitat in relation to external factors such as climate changes that can severely affect ecosystems.

Quality assurance of ethical issues and regulatory aspects relating to good clinical practices in the HELENA Cross-Sectional Study.

RATIONALE: Research involving humans is regulated by regulatory authorities through their specific requirements and controls. The Healthy Life Style in Europe by Nutrition in Adolescence Cross-Sectional Study (HELENA-CSS) is a multicenter biomedical research study of adolescents in several representative European cities, which requires satisfying medico-regulatory requirements including Independent Ethics Committee (IEC) approval and agreement by the national or local regulatory authorities. To achieve a high level of quality assurance relating to ethical issues, we followed the good clinical practices (GCP) described at the International Conference on Harmonisation (ICH), which we adapted to the national and local situations of each of the 11 participating cities in 10 European countries. OBJECTIVE: The main objective of the HELENA-CSS is to evaluate reliable and comparable data of nutritional habits and lifestyle in a representative sample of European adolescents. The aim of this paper is to present the methods relating to the ethical and regulatory issues of this study and to describe the current state of the medico-regulatory requirements involved in conducting this kind of study in each country. MATERIALS AND METHODS: Following the GCP-ICH guidelines, a protocol describing the HELENA-CSS was written and approved by all partners. In the pilot study, a case report form adapted to the study objectives and its manual of operation was constructed and used by all partners. All information letters to adolescents and their parents and consent forms were first written in English, then translated into the local language, and adapted to each local situation. All documents were then checked centrally for any deviation and corrected if required. An operation manual relating to ethical issues and other medico-regulatory requirements was also developed. This paper presents the current status of the medico-regulatory requirements from each HELENA-CSS participant country. RESULTS: Before the beginning of the study, most centers had satisfied the medico-regulatory requirements of IEC approval and agreement with other national or local regulatory authorities/organizations. For a few centers, some problems were detected and corrective actions were taken to improve missing information to reach a high level of quality assurance of ethical issues. CONCLUSION: The GCP-ICH guidelines about nontherapeutic biomedical research are interpreted and applied differently across Europe. This study shows that high-quality nontherapeutic biomedical research can address the ethical issues included in the GCP-ICH regulations and can be harmonized among the HELENA European partners.

Socioeconomic questionnaire and clinical assessment in the HELENA Cross-Sectional Study: methodology.

RATIONALE: Environmental factors such as dietary habits, breastfeeding, socioeconomic conditions and educational factors are strong influences on nutritional and puberty status, physical activity, food choices and their interactions. Several diseases of adulthood seem to be linked to, or to originate from, lifestyle in childhood and adolescence. OBJECTIVE: The aims of this study are to describe birth parameters and socioeconomic factors and to assess clinical status in adolescents aged 13-16 years from 10 European countries participating in the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) Cross-Sectional Study (CSS). METHODOLOGY: A self-report questionnaire on the socioeconomic status, a parental questionnaire concerning neonatal period and also a case report form (CRF), in which clinical items during clinical examination (such as medical history, treatments, anthropometry, Tanner staging, blood pressure, heart rate) were assessed. To develop these documents, first a list of items was established, a search of existing documents was performed and the advice of local and international experts was taken. All documents (questionnaires and an operations manual) were discussed in plenary HELENA meetings; a final version of these documents was fixed, and the process of translation and back translation was performed. RESULTS: The questionnaires and CRF were tested for validation in all 10 participant cities; 208 adolescents were enrolled during the pilot study. All items that caused problems or questions in one or more participating centers or were completed by < 85% of the adolescents were reviewed before the beginning of the HELENA-CSS. CONCLUSION: These final questionnaires and CRF will contribute to better understanding of the inequalities in nutrition, behavior and health in the European adolescent population. The experience and process should be useful for other multicenter studies.

Design and implementation of the Healthy Lifestyle in Europe by Nutrition in Adolescence Cross-Sectional Study.

OBJECTIVE: To provide an overview of the Healthy Lifestyle in Europe by Nutrition in Adolescence Cross-Sectional Study (HELENA-CSS) design, with particular attention to its quality control procedures. Other important methodological aspects are described in detail throughout this supplement. DESIGN: Description of the HELENA-CSS sampling and recruitment approaches, standardization and harmonization processes, data collection and analysis strategies and quality control activities. RESULTS: The HELENA-CSS is a multi-centre collaborative study conducted in European adolescents located in urban settings. The data management systems, quality assurance monitoring activities, standardized manuals of operating procedures and training and study management are addressed in this paper. Various quality controls to ensure collection of valid and reliable data will be discussed in this supplement, as well as quantitative estimates of measurement error. CONCLUSION: The great advantage of the HELENA-CSS is the strict standardization of the fieldwork and the blood analyses, which precludes to a great extent the kind of immeasurable confounding bias that often interferes when comparing results from isolated studies.

Characterisation of bioactive compounds in infant formulas using immobilised recombinant estrogen receptor-alpha affinity columns.

In this study, the use of recombinant estrogen receptor alpha (ERalpha)-based affinity columns was reported, for the isolation and the identification of estrogenic substances present in complex matrices, focusing on bioactive compounds present in foodstuff. The capability of affinity columns to trap high, but also low-affinity radio-labelled ligands (17beta-estradiol, genistein and bisphenol A) was demonstrated. Three pooled samples of infant formulas (milk-based, hypoallergenic and soy-based formulas for infants aged 0-4 months) from a EU market basket were prepared by the CASCADE Network of Excellence. After determining the estrogenic activity of these food samples, human recombinant ERalpha ligand binding domain (LBD) based affinity columns combined with suitable analytical methods (high resolution LC-MS/MS) were used to identify the bioactive compounds present in the soy-based formula extract, namely phytoestrogens (genistein and daidzein) involved in the agonistic activity measured. Incubations of genistein with liver microsomes were carried out and the extracts analysed following the same protocol, demonstrating that hERalpha affinity columns can also be used for trapping active metabolites. This approach combining bioluminescent cell lines with this useful tool based on hERalpha-LBD affinity columns thus allowed the purification and the concentration of both known and unknown estrogenic ligands prior to investigation of their structure using LC-MS.