Sarphati Amsterdam: a dynamic research infrastructure

Sarphati Amsterdam is a collaboration between the city of Amsterdam and four Amsterdam research institutions, focusing on innovative multidisciplinary research beneficial to preventing non-communicable diseases effectively and sustainably. With the Sarphati Cohort, Sarphati Amsterdam has established a dynamic cohort study that systematically monitors growth and its determinants from birth until adulthood, in order to 1) identify causes of overweight and 2) evaluate interventions to combat overweight. Data collection is linked to routine Youth Health Care consultations with all ∼150,000 Amsterdam children (0-18y). Thus, making selected care data on growth and development available for research. After informed parental consent, anonymised data can be used to systematically monitor children. Additional data is collected through age-specific questionnaires. The sub-cohort structure facilitates more extensive data collection in subpopulations, including interviews, observations, and biosamples. Up until March 2020 over 5,130 children enrolled. The continuous inclusion of new-borns gives the Sarphati Cohort its dynamic character. This enables the incorporation of innovative research designs to evaluate interventions.With over 170 different nationalities Amsterdam has a highly diverse population, both in terms of cultural and socio-economic background. The data collection strategies and tailored tools are developed to meet the level of adaptability this design requires. Embedding research in this manner enriches it way beyond clinical settings and provides an unparalleled depth and breadth of data. With the Sarphati Cohort, Sarphati Amsterdam facilitates innovative research in the field of obesity that will contribute to the ambitious policy objectives of the city of Amsterdam to promote healthy behaviour and improve the quality of life of young people.

MPC-09 THE OPTIMIZATION OF TREATMENTS FOR SO-CALLED PRIMITIVE NEUROECTODERMAL TUMORS WITH MOLECULAR ANALYSIS

INTRODUCTION In the previous WHO classification of central nervous tumors, the supratentorial tumors comprise small round blue cells with aggressive clinical features had been defined as primitive neuroectodermal tumor (PNET). Recent molecular analysis revealed that they do not belong to a single entity, but they are re-classified as the tumors of other well-defined tumors and newly defined tumor species. These facts were reflected to the new classifications. While, there are few studies those showed the re-consideration of treatments for tumors diagnosed as so-called PNET. In this study, we propose the optimization of treatments for tumors diagnosed by the new classification to clarify which treatments were effective for the tumors those were diagnosed as PNET. MATERIALS AND METHODS The tumor samples diagnosed as so-called PNETs were analyzed. The molecular information was extracted from tumor specimens. We used high throughput analysis with microarray, FISH, and immunohistochemistry. They all had treated in our institution in last 6 years and their clinical courses were followed by medical records. Informed parental consent was obtained from their guardians and this study was approved by the institutional review board of Juntendo university. RESULTS Nine tumor samples were able to be analyzed and they are re-classified into high-grade glioma, neuroblastoma, sarcoma, embryonal tumors with multilayered rosettes, C19MC altered (ETMR). They resembled each other closely in morphology, and therefore, it was not able to be classified by histopathological findings. There was a case of pineoblastoma, whose molecular background suggested that the tumor was re-classified into neuroblastoma. In terms of treatments, we have succeeded in neuroblastoma cases so far, ETMRs were required multiple surgeries and radiations to maintain remissions. CONCLUSIONS Re-classification of diagnosis based on the molecular background is necessary to clarify the optimization of treatments for pediatric brain tumors, and the comprehensive methods is required. We present our methods for molecular diagnosis in clinical field and future plans.

Comparison of IFN-γ Levels in Children with Tuberculosis Disease (TB) and Latent Tuberculosis Infection (LTBI)

AIM: This study aimed to evaluate the importance of IFN-γ in the diagnosis of pediatric TB and LTBI and to compare the IFN-γ levels. METHODS: We analysed 100 patients examined for possible M. tuberculosis infection or disease at the Institute of Respiratory Diseases in Children, Kozle, Skopje. Patients were divided into 2 groups: TB disease and LTBI. The following parameters were analyzed: demographic characteristics, history of previous exposure to active TB, BCG vaccination and presence of BCG scar, lung X-ray findings, tuberculin skin test by the Monteux method and the value of INF -γ according to the Quantiferon TB gold test, direct samples of acid-alcohol-resistant bacilli of sputum and Löwenstein Jensen cultures. Informed parental consent was obtained for each child included in the study. RESULTS: In the LTBI group 60.9% had a scar from the vaccination while in the TB group 50% had BCG scar. TST induration diameters in children with or without BCG scar were significantly larger in patients with active TB. Children with active TB had significantly higher IFN-γ levels than children with LTBI. The IFN-γ for the cut-off of 0.35 IU/ml, has 64% sensitivity for detection of LTBI, versus 80.6% sensitivity for active disease. Children with close TB contact had significantly higher IFN-γ levels. Correlation between TST induration diameter and IFN-γ levels was stronger in the TB group. CONCLUSION: IFN-γ levels are significantly higher in children with active TB, and children with close contact with TB patient. It has better sensitivity in active TB. Using both tests (IFN-γ and TST) can improve the diagnose of LTBI and TB in countries where vaccination with BCG is widespread.

Improved Measurement of Minimal Residual Disease (MRD).

A new and improved method for measurement of minimal residual disease (MRD) was developed. Method the total repertoire of leukaemic rearrangements of the immunoglobulin heavy chain (IgH) gene.was determined by performing multiple parallel Q-PCRs in microplates to determine usage of individual V and J segments. This enabled detection and quantification of clones ranging in size from 100% to approximately 0.03% of the leukemic population. MRD measurement involved nested Q-PCR using the specific V and J primers and internal primers based on the sequence of the rearrangement of interest. Following informed parental consent, 25 children with B-ALL were studied at the end of induction therapy. Under general anesthesia, 4 aspirations, 2 from each iliac spine, were performed and on each sample MRD was measured on 2 different days. This enabled definition of the laboratory and sampling factors important in measurement of MRD. Results Repertoire analysis was very effective in identifying rearrangements of the IgH gene suitable for use as molecular markers, being superior to the commonly-used BIOMED-2 protocol for identification of both large and small clones. Two or more rearrangements marking large clones were detected in 20 of the 25 patients. The median MRD level at the end of induction was 2.1 x 10−5. A level of > 10−3 was seen in 4 patients and < 10−7 in 4. Sensitivity of detection in a single sample was approximately 2 x 10−6, which is 1–2 orders of magnitude better than current techniques. Figure Figure The SD of measurement depended on the number of target rearrangements present in the sample, reflecting the Poisson statistical uncertainty inherent in measurement of small numbers of events. For > 50 rearrangements SD was 0.23 log units, but below this level the SD rose steeply. 50 targets in 1 μg of DNA corresponds to an MRD of approximately 3 x 10−4. Figure Figure There was significant sampling error. In 1 patient there was 1000-fold MRD difference between the 2 iliac spines. Conclusions We recommend that, for MRD measurement, -an aspiration should be performed from each iliac spine, with each sample being quantified separately and an average obtained -each measurement should involve at least 10 μg of good-quality DNA The MRD value so obtained should have sufficient accuracy, sensitivity and precision for clinical decisions based upon the value to be made with confidence. The described methods for MRD measurement should also be applicable to monitoring of all B-lymphocyte neoplasms, in addition to ALL.

Effect of anaesthetic agents on tympanometry and middle-ear effusions

Following informed parental consent 93 children underwent bilateral grommet insertion. Tympanometry was performed pre-operatively, and immediately prior to myringotomy. A standardized anaesthetic was used. At myringotomy the presence or absence of fluid was recorded, as well as the time since induction of the general anaesthetic.A pre-operative type B tympanogram predicted a middle-ear effusion at myringotomy in 92 per cent of patients. A pre-operative type C2 tympanogram predicted a middle-ear effusion at myringotomy in 39 per cent of patients. Sixty tympanograms (30 per cent) changed following a general anaesthetic. Fourteen type B tympanograms changed to type A and eight of these had effusions. The duration of the general anaesthetic did not influence the probability of a middle-ear effusion being present at myringotomy. A pre-operative type B tympanogram is a good predictor of middle-ear fluid. The duration of the general anaesthetic is not significant in predicting the presence of a middle-ear effusion.

Single-breath Vital Capacity Rapid Inhalation Induction in Children

Background The authors compared the speed of induction of anesthesia with sevoflurane with and without nitrous oxide with the speed of halothane and nitrous oxide using a single-breath vital capacity induction. Methods With informed parental consent, 51 healthy unpremedicated children aged 5-12 yr were randomized to inhale a single breath of one of three gas mixtures: 8% sevoflurane in 66% nitrous oxide, 8% sevoflurane in oxygen, or 5% halothane in 66% nitrous oxide. A blinded observer recorded the times to loss of the eyelash reflex, return of conjugate gaze, the presence of airway reflex responses, involuntary movement, and hemodynamic responses. Results Forty-two children completed the study. The times (mean +/- SD) to loss of the eyelash reflex with sevoflurane/nitrous oxide, 38+/-8 s, and for sevoflurane-oxygen, 34+/-12 s, were less than that with halothane-nitrous oxide, 58+/-17 s (P &lt; 0.01). Movement occurred less frequently during sevoflurane than during halothane anesthesia (P &lt; 0.05). The times to return of conjugate gaze and the incidence of airway reflex responses were similar among the groups. The incidence of dysrhythmias in the sevoflurane groups was less than that in the halothane group (P &lt; 0.01). Conclusions Induction of anesthesia with a single breath of 8% sevoflurane with or without 66% nitrous oxide is more rapid than with 5% inspired halothane with 66% nitrous oxide in children. The incidence of movement and dysrhythmias during a single-breath induction with sevoflurane are less than they are with halothane.

Half-Dose Immunization for Diphtheria, Tetanus, Pertussis

In Reply.— We appreciate your comments regarding our article in which we reported on the response of preterm infants to half-dose diphtheria, tetanus, pertussis immunizations. Although your points are well taken, it would be difficult logistically to carry out a study in which the serologic responses to 6 (or more) doses of half-dose vaccine are measured. At the very least, it would be hard to obtain informed parental consent. In our study, we attempted to show how poor the serologic response was to a reduced dosage of vaccine.

Effect of Informed Parental Consent on Mothers' Knowledge of Newborn Screening

To determine whether knowledge was improved as a result of obtaining informed consent from parents for newborn screening of their infants for phenylketonuria (PKU) and other hereditary metabolic disorders, new mothers in seven Maryland hospitals were interviewed either before receiving a standard disclosure (n = 210) or after giving consent (n = 418). The mean knowledge score of the women interviewed after giving consent was significantly higher (P &lt; .001). Receiving the disclosure was a more powerful predictor of knowledge score, accounting for 40% of the variance, than demographic factors, which accounted for 9%. Women whose consent was obtained just prior to discharge tended to have lower knowledge scores than women whose consent was obtained earlier (P = .03). Women with higher knowledge scores were somewhat less likely to favor consent than women with lower scores. Although consent may not be appropriate for some low-risk procedures, informing parents can be easily and inexpensively accomplished.