Prenatal metal mixtures and child blood pressure in the Rhea mother-child cohort in Greece

Background: Child blood pressure (BP) is predictive of future cardiovascular risk. Prenatal exposure to metals has been associated with higher BP in childhood, but most studies have evaluated elements individually and measured BP at a single time point. We investigated impacts of prenatal metal mixture exposures on longitudinal changes in BP during childhood and elevated BP at 11 years of age. Methods: The current study included 176 mother-child pairs from the Rhea Study in Heraklion, Greece and focused on eight elements (antimony, arsenic, cadmium, cobalt, lead, magnesium, molybdenum, selenium) measured in maternal urine samples collected during pregnancy (median gestational age at collection: 12 weeks). BP was measured at approximately 4, 6, and 11 years of age. Covariate-adjusted Bayesian Varying Coefficient Kernel Machine Regression and Bayesian Kernel Machine Regression (BKMR) were used to evaluate metal mixture impacts on baseline and longitudinal changes in BP (from ages 4 to 11) and the development of elevated BP at age 11, respectively. BKMR results were compared using static versus percentile-based cutoffs to define elevated BP. Results: Molybdenum and lead were the mixture components most consistently associated with BP. J-shaped relationships were observed between molybdenum and both systolic and diastolic BP at age 4. Similar associations were identified for both molybdenum and lead in relation to elevated BP at age 11. For molybdenum concentrations above the inflection points (~ 40–80 μg/L), positive associations with BP at age 4 were stronger at high levels of lead. Lead was positively associated with BP measures at age 4, but only at high levels of molybdenum. Potential interactions between molybdenum and lead were also identified for BP at age 11, but were sensitive to the cutoffs used to define elevated BP. Conclusions: Prenatal exposure to high levels of molybdenum and lead, particularly in combination, may contribute to higher BP at age 4. These early effects appear to persist throughout childhood, contributing to elevated BP in adolescence. Future studies are needed to identify the major sources of molybdenum and lead in this population

Ανίχνευση του οροεπιπολασμού των polyomaviruses, herpesviruses & helicobacter pylori σε πρώιμες ηλικίες και η συσχέτισή του με επιπτώσεις στην υγεία των παιδιών: προοπτική μελέτη μητέρας-παιδιού Κρήτης, μελέτη ΡΕΑ

Introduction: During the first years of life, children are constantly confronted to a number of infections. Clinically evident infections have been a subject of research for many years, enabling development of successful prevention programs (e.g., vaccination programs). Nonetheless, children subclinically acquire a number of other infections whose role in health and disease has been little examined. It has been suggested that a normal course of infections in early life protects against the development of childhood leukemia (the Greaves hypothesis) and of chronic inflammatory disorders (the hygiene or “old friends” hypothesis). Nonetheless, these hypotheses are based on epidemiological studies which use proxy variables such as age of first school attendance, to denote exposure to common infections. Very limited prospective data exist on characterizing this repertoire of common infections in childhood by use of biological measures.The primary objective of this study has been the description of the seroepidemiology of Polyomaviruses and Herpesviruses in early childhood. Polyomaviruses are DNA viruses (fourteen known so far), which are largely acquired subclinically in early life. Studies in adults show high seroprevalence rates but the exact age of acquisition is not known. Herpesviruses are also largely acquired in childhood with the exception of HSV-2, which is predominantly acquired through sexual contact later in life. Emerging data suggest that recent changes in family structure, childrearing, and hygiene practices have modified the epidemiology of Herpesviruses. This might have important public health implications as the timing of exposure has a critical role in the outcome of infection with Ηerpesviruses. These facts highlight the importance of studying the seroepidemiology of Polyomaviruses and Herpesviruses in early life in the general population.The next objective has been to explore the role of Polyomaviruses and Herpesviruses infections in child’s neurodevelopment and growth. The developing brain is extremely sensitive to the neuroinflammation induced by viruses. Among Polyomaviruses, JCPyV is known to be neurotropic and is the etiologic agent of progressive multifocal leukoencephalopathy, a fatal demyelinating disease of the central nervous sytem occurring mainly in immonocompromised individuals. Moreover, Lintas et al. foundenhanced frequencies of Polyomaviruses (BKPyV, JCPyV, SV40) in postmortem neocortical tissues of autistic patients, raising questions for an association between Polyomaviruses and autism. EBV, CMV and HSV-1, members of the Herpesviruses family, are also neurotropic viruses and under certain circumstances can infect the central nervous system and cause acute encephalitis/meningitis and long-term neurological sequelae (e.g., sensorineural hearing loss due to congenital CMV infection). The potential association of Polyomaviruses and Herpesviruses with child’s growth, obesity and cardiometabolic traits is less evident but is based on an emerging hypothesis of an infectious origin of obesity. In particular, adenovirus 36 has been associated with obesity both in animal and human studies. Moreover a metabolic dysfunction has been shown to accompany some viral infections such as CMV, HSV-1 and HSV-2. Interestingly, an increasing number of studies report a link between pathogen burden, obesity, cardiometabolic traits and disease, but no evidence exists in childhood. Lastly, an objective of the present study has been to describe the seroepidemiology of Helicobacter pylori, a very common chronic bacterial infection in humans which is associated with increased risk for gastric cancer. A wealth of evidence strongly implicates Helicobacter pylori infection in the pathogenesis of extragastric diseases such as idiopathic thrombocytopenic purpura, unexplained iron-deficiency anemia and vitamin B12 deficiency. Moreover, an increasing number of studies report an association between Helicobacter pylori and neuropsychiatric diseases such as Parkinon’s and Alzheimer. It is likely that an association with neurodevelopment is evident already in early life, but there are no data. Thus, we hypothesized that Helicobacter pylori infection during the first years of life but also maternal Helicobacter pylori infection during pregnancy may unfavorably affect offspring’s neurodevelopment through micronutrients deficiencies (e.g folate, B12). Fetal brain is extremely vulnerable to such micronutrients defects. To the best of our knowledge, no study has examined differences in neurodevelopment between children of Helicobacter pylori seropositive versus seronegative mothers.Specific Objectives: • To describe the seroprevalence of Polyomaviruses, Herpesviruses and Helicobacter pylori in early life using repeated samples collected at birth (cord blood), three years and four years of age. • To identify the factors – sociodemographic, perinatal, variables denoting social interactions, lifestyle, hygiene practices- that determine the acquisition of Polyomaviruses and Herpesviruses up to four years of age. • To explore the association between seropositivity to single and multiple Polyomaviruses and Herpesviruses up to age four with neurodevelopmental assessment of children at the same age.• To explore the association between Helicobacter pylori infection early in life including fetal life with neurodevelopment of children at age four.• To explore the association between seropositivity to single and to multiple Polyomaviruses and Herpesviruses up to age four with obesity and cardiometabolic traits at age four and six.Methods: This study uses data from the Rhea study a birth cohort that recruited 1,606 pregnant women from February 2007 to January 2008 (www.rhea.gr). The inclusion criteria were confirmed pregnancy, residency within the study area, aged 16 years or older, and good understanding of the Greek language. Recruitment occurred mostly before 15 weeks of gestation. Mothers were contacted again at 24 weeks of gestation, at birth, and for child’s follow-up at 9th month, 18th month, 4 years, and 6 years of age. When members of the cohort were 3 years of age, we invited a randomly selected subsample for a serological assessment (n = 109).Blood samples collected at birth (cord blood) and at 3 and 4 years of age were processed following standard procedures and then stored at −80°C. Serum aliquots of 100 μL were shipped on dry ice to the German Cancer Research Center, Heidelberg, Germany, for serological analysis. Immunoglobulin G seroreactivity against the viral capsid protein 1 of ten Polyomaviruses (BKPyV, JCPyV, KIPyV,WUPyV,MCPyV, HPyV6, HPyV7, TSPyV, HPyV9, HPyV10), four EBV antigens (ZEBRA, EBNA-1, EA-D, VCA p18), five CMV antigens (pp52, pp65, pp150, pp28, CM2), one HSV-1 antigen (gG), one HSV-2 antigen (mgGunique), four HHV-8 antigens (LANA, v-cyclin, K8.1, ORF-65) and twelve H.pylori proteins (GroEL, UreA, HP0231, NapA, HP0305, HpaA, CagA, HyuA, catalase, VacA, HcpC, Omp) was measured by fluorescent bead-based multiplex serology (1:1,000 dilution). Of note, immunoglobulin G levels in cord blood reflect the mother’s immunoglobulin G levels. Seropositivity for a given antigen was defined as median fluorescence intensity values greater than the antigen- specific cut off. EBV and CMV seropositivity was defined as seropositivity for at least two virus specific antigens. Seropositivity for a given Helicobacter pylori protein was defined as seroreactivity greater than the protein-specific cut-off which was re-evaluated using the published reference panel. The aggregate number of different Polyomaviruses that the children were seropositive to was referred as Polyomaviruses burden. Similarly, we calculated the Herpesviruses burden.Information on potential determinants of seroprevalence of polyomaviruses and herpesviruses is based on repeated questionnaires from birth to 4 years of age. Children’s cognitive and motor development was assessed by two trained psychologists with the McCarthy Scales of Children’s Abilities (MSCA). Additional information on children’s behavior was obtained via maternal report on standardized child behavior scales, which were administered at the 4 years of age follow-up. The Attention-Deficit/Hyperactivity Disorder Test (ADHDT) is designed to identify and evaluate ADHD in ages 3–23 years. The parent version of the Strengths and Difficulties Questionnaire (SDQ) is a behavioral screening instrument designed to assess strengths and difficulties of children aged 3–16 years. We measured body mass index, waist circumference and skinfold thickness at four anatomical sites at four and six years of age. At age six hand-to-leg bioelectrical impendence was also measured using a Bodystat 1500 machine. Data on metabolic traits including serum lipids, leptin and adiponectin levels were also available.Descriptive analyses of the study population characteristics, exposures and outcomes were conducted. Multiple linear, logistic and Poisson regression models were used to explore the associations accordingly.Results: 1) Seroprevalence of Polyomaviruses ranged from 38.5% to 99.8% in cord blood and from 20.9% to 82.3% at age 4. Seroprevalence of EBV, CMV, HSV-1, HSV-2 and HHV-8 was 99.4%, 74.9%, 26.2%, 8.0%, and 1.6% in cord blood and 52.5%, 25.8%, 3.6%, 1.4%, and 0% at age 4, respectively. Determinants of seropositivity at age 4 were cord seropositivity (JCPyV, HPyV7, HPyV10, CMV), vaginal delivery (HPyV10), breastfeeding (CMV), younger age at day-care entry (BKPyV, KIPyV, WUPyV, TSPyV, HPyV10, HPyV9, EBV, CMV), and swimming pool attendance (BKPyV, KIPyV, WUPyV, HPyV10). Televisionviewing, parental stress, and hygiene practices were inversely associated with the seroprevalence of Polyomaviruses and Herpesviruses.2) Based on crosss-ectional data at age four, we investigated the association between seropositivity to single or multiple Polyomaviruses and Herpesviruses with child’s neurodevelopment and behavior among 674 children. Seropositivity to BKPyV, a potential neurotropic virus, was associated with higher score in ADHDT inattention subscale. Moreover, children seropositive to ≥8 polyomaviruses had lower score in ADHDT inattention subscale and lower score in SDQ hyperactivity–inattention subscale versus children seropositive to ≤3 Polyomaviruses.3) H.pylori seroprevalence in cord blood, representing maternal status, was 41.5% and in children four years of age was 6.5%. Children of Helicobacter pylori seropositive mothers compared to those of seronegative mothers, had lower score in all scales of the MCSA excluding motor scale. In addition, seropositivity in cord blood specifically to GroEl and NapA – two of the 12 Helicobacter pylori proteins investigated – was associated with statistically significant lower scores in almost all scales of MCSA. At age four, Helicobacter pylori seropositive children performed worst in neurodevelopment assessment compared to their seronegative counterparts although no association reached statistically significant level.4) Based on cross-sectional data of 674 children participating at the 4 years of age follow-up and prospective data of 440 children at age six, we investigated the association of seropositivity to single or multiple Polyomaviruses and Herpesviruses with child’s obesity and cardiometabolic traits at the corresponding ages. BKPyV seropositivity was associated with lower BMI, waist circumference, sum of skinfolds and lower leptin levels at age four and with lower BMI, waist circumference and %body fat at age six. On the other hand, CMV seropositivity was associated with higher BMI at age four and six and higher sum of skinfolds at age six. Moreover, children with “2-3 Herpesviruses infections” versus those with “0 Herpesviruses infections” had higher BMI SD-score, waist circumference and sum of skinfolds at age four.Conclusions: In summary, findings of the present thesis describe the natural history of acquisition of ten Polyomaviruses (BKPyV, JCPyV, KIPyV, WUPyV, HPyV6, HPyV7, TSPyV, MCPyV, HPyV9 and HPyV10), five Herpesviruses (EBV, CMV, HSV-1, HSV-2, HHV-8) and Helicobacter pylori in a population of young healthy children which are prospectively followed in the Rhea mother-child cohort in Crete. To the best of our knowledge, there is no previous report gathering such detailed information on seroprevalence to those common infections. We have shown that, by age 4, children were commonly seropositive to Polyomaviruses and Herpesviruses, with the exceptions of HHV-8, HSV-1 and HSV-2. Seroprevalence to Helicobacter pylori was low and in accordance with reports from other western populations. The resent analysis provides important insights into the factors that determine the acquisition of such common infections by age four. For example, younger age at day-care entry importantly contributed in the epidemiology of BKPyV, KIPyV, WUPyV, TSPyV, HPyV10, HPyV9,EBV, and CMV while there was as strong socioeconomic position patterning of Polyomaviruses seroprevalence at age four. Such findings are important allowing inference on potential routes of transmission for those viruses among young children.Intriguing are the findings regarding the association of Polyomaviruses and Herpesviruses infections with child’s neurodevelopment. The inverse association between the number of Polyomaviruses infections acquired up to age four with symptoms of inattention and hyperactivity might indicate not only potential interaction between the immune system and the developing brain but also a different pattern of acquisition of such common infections among children with such behaviours. From a public health perspective, it is particularly important the finding that children of Helicobacter pylori seropositive mothers had worst neurodevelopmental performance at age four, raising questions on whether treatment for Helicobacter pylori could prevent such an outcome. Finally, we observed very strong associations for BKPyV and CMV with obesity and cardiometabolic traits in childhood. Similar findings have not been reported in children and should be replicated in other populations.Εισαγωγή: Τα πρώτα χρόνια της ζωής, τα παιδιά εκτίθενται συνεχώς σε λοιμώξεις. Οι λοιμώξεις που συνοδεύονται από κλινικές εκδηλώσεις έχουν μελετηθεί συστηματικά και έχουν οδηγήσει στην ανάπτυξη αποτελεσματικών μέτρων πρόληψης, όπως ο εμβολιασμός. Ωστόσο, ο ρόλος των κοινών λοιμώξεων στην υγεία και τη νόσο έχει ελάχιστα μελετηθεί. Υπάρχουν υποθέσεις που υποστηρίζουν ότι μια φυσιολογική πορεία έκθεσης σε λοιμώξεις στην παιδική ηλικία αποτελεί προστατευτικό παράγοντα για την ανάπτυξη της παιδικής λευχαιμίας (η υπόθεση του Greaves) καθώς και για την ανάπτυξη χρόνιων φλεγμονώδων νοσημάτων (η υπόθεση της υγιεινής). Παρόλα αυτά οι υποθέσεις αυτές βασίζονται σε μελέτες που η έκθεση σε κοινές λοιμώξεις εκφράζεται με μεταβλητές όπως η ηλικία έναρξης του σχολείου. Ελάχιστες προοπτικές μελέτες υπάρχουν που να περιγράφουν τη φυσική πορεία έκθεσης σε πολλές κοινές λοιμώξεις ταυτόχρονα τα πρώτα χρόνια της ζωής, χρησιμοποιώντας βιολογικούς δείκτες, για παράδειγμα μετρώντας την ανοσολογική απάντηση.Αντικείμενο της παρούσας διατριβής αποτέλεσε αρχικά η περιγραφή της έκθεση στην παιδική ηλικία στους Polyomaviruses και Herpesviruses. Οι Polyomaviruses αποτελούν μια ομάδα DNA ιων που προσβάλουν τον άνθρωπο (δεκατέσσερις γνωστοί μέχρι σήμερα) και φαίνεται να αποκτώνται νωρίς στη ζωή, προκαλώντας υποκλινική νόσο. Μελέτες σε ενήλικες έχουν δείξει πολύ υψηλά ποσοστά οροεπιπολασμού ωστόσο δεν είναι γνωστή η συνήθης ηλικία ορομετατροπής. Οι Herpesviruses αποκτώνται συνήθως στην παιδική ηλικία με εξαίρεση τον HSV-2 που αποκτάται αργότερα στη νεαρή ενήλικο ζωή με την έναρξη της σεξουαλικής δραστηριότητας. Νεότερα δεδομένα δείχνουν ότι η επιδημιολογία των Herpesviruses έχει αλλάξει σημαντικά και η ορομετατροπή έχει μετατεθεί σε μεγαλύτερες ηλικίες, πιθανά λόγω αλλαγών που έχουν επισυμβεί στον τρόπο ζωής όπως η σύνθεση της οικογένειας και οι τρόποι ανατροφής των παιδιών. Τα παραπάνω υποσημαίνουν την αξία μελέτης του οροεπιπολασμού των Polyomaviruses και Herpesviruses τα πρώτα χρόνια ζωής στο γενικό πληθυσμό. Επόμενο σημαντικό ερώτημα της παρούσας διατριβής αποτέλεσε κατά πόσο οι Polyomaviruses και Herpesviruses σχετίζονται με την νευροανάπτυξη και την σωματική ανάπτυξη στα πρώτα χρόνια ζωής στο γενικό πληθυσμό. Το αναπτυσσόμενο νευρικό σύστημα είναι ιδιαίτερα ευαίσθητο στην φλεγμονή που προκαλείται απο ιογενείς λοιμώξεις. Από τους Polyomaviruses, o JCPyV είναι νευροτρόπος ιός και ευθύνεται για την προοδευτική πολυεστιακή λευκοεγκεφαλοπάθεια σε ανοσοκατεσταλμένα άτομα. Επίσης σε μία μελέτη ασθενών-μαρτύρων, απομονώθηκαν οι BKPyV, JCPyV και SV40 σε εγκεφαλικό ιστό ατόμων με αυτισμό. Από τους Herpesviruses που μελετήσαμε, οι EBV, CMV και HSV-1 είναι δυνητικά νευροτρόποι ιοί και κάτω από ορισμένες συνθήκες μπορούν να οδηγήσουν σε εγκεφαλίτιδα/μηνιγγίτιδα καθώς και μακροπρόσθεσμες νευρολογικές επιπτώσεις, για παράδειγμα σε νευροαισθητήρια απώλεια της ακοής μετά από συγγενή λοίμωξη με CMV. Η πιθανή σχέση των Polyomaviruses και Herpesviruses με την σωματική ανάπτυξη, την παχυσαρκία και καρδιαγγειακούς δείκτες στην παιδική ηλικία είναι λιγότερο προφανής, ωστόσο στηρίζεται σε μια νεότερη υπόθεση ότι οι λοιμώξεις μπορεί να οδηγούν σε παχυσαρκία μέσω διαφόρων μηχανισμών που υποδεικνύονται από μελέτες σε ζώα. Συγκεκριμένα, ο αδενοϊός 36, που αποκτάται στην παιδική ηλικία, έχει συσχετιστεί με την παχυσαρκία σε μελέτες σε ανθρώπους και ζώα. Επίσης υπάρχουν αναφορές για τη σχέση των CMV, HSV-1, HSV-2 με καρδιαγγειακούς δείκτες ωστόσο τα δεδομένα αυτά είναι περιορισμένα. Τα τελευταία χρόνια αυξάνονται επίσης οι αναφορές για τη σχέση του αριθμού των λοιμώξεων που έχει εκτεθεί ο άνθρωπος (pathogen burden) με την παχυσαρκία και άλλους καρδιαγγειακούς δείκτες, ωστόσο δεν υπάρχουν δεδομένα στην παιδική ηλικία. Τέλος, αντικείμενο της παρούσας διατριβής αποτέλεσε ο καθορισμός του οροεπιπολασμού του Helicobacter pylori, ενός κοινού παθογόνου για τον άνθρωπο που σχετίζεται με αυξημένο κίνδυνο για γαστρικό καρκίνο. Ιδιαίτερο ενδιαφέρον παρουσιάζουν οι αυξανόμενες αναφορές για τη σχέση του Helicobacter pylori με εξωγαστρικές εκδηλώσεις όπως η ιδιοπαθής θρομβοπενική πορφύρα, η σιδηροπενική αναιμία και η Β12 ανεπάρκεια. Παράλληλα, υπάρχουν αναφορές σε ενήλικες που συσχετίζουν το Helicobacter pylori με νευροψυχιατρικά νοσήματα όπως η νόσος Alzheimer και Parkinson. Πιθανά παρόμοια συσχέτιση να υπάρχει και στην παιδική και εφηβική ηλικία ωστόσο δεν υπάρχουν σχετικές αναφορές. Υποθέσαμε ότι η λοίμωξη από Helicobacter pylori νωρις στη ζωή, ακόμα και κατά την ενδομήτριο ζωή, μπορεί να οδηγήσει σε δυσμενή νευροαναπτυξιακή εξέλιξη, μέσω μικροθρεπτικών ελλείψεων (π.χ. σε φυλλικό, B12) που φαίνεται να προκαλεί η λοίμωξη από Helicobacter pylori. Στην παρούσα διατριβή μελετήθηκε λοιπόνσυγκεκριμένα η σχέση της οροθετικότητας στο Helicobacter pylori στις μητέρες στο τέλος της κύησης και των παιδιών στα 4 έτη με τη νευροαναπτυξη των παιδιών στα 4 έτη.Ειδικοί Στόχοι: • Να περιγράψουμε τον οροεπιπολασμό των Polyomaviruses, Herpesviruses και Helicobacter pylori νωρίς στη ζωή, χρησιμοποιώντας επαναλαμβανόμενα δείγματα αίματος που συλλέχθησαν στη γέννηση (ομφαλικό αίμα), στα 3 έτη και στα 4 έτη.• Να περιγράψουμε τους παράγοντες- κοινωνικο-οικονομικούς, περιγεννητικούς, κοινωνικών επαφών, τρόπου ζωής, προσωπικής υγιεινής- που επιδρούν στην ορομετατροπή στους Polyomaviruses και Herpesviruses εως την ηλικία των 4 ετών.• Να μελετήσουμε τη σχέση της έκθεσης σε ένα ή και περισσότερους Polyomaviruses και Herpesviruses εως την ηλικία των 4 ετών με τη νευροαναπτυξη των παιδιών στην ίδια ηλικία.• Να μελετήσουμε τη σχέση της λοίμωξης με Helicobacter pylori νωρίς στη ζωή, συμπεριλαμβανομένου της ενδομήτριου ζωής, με τη νευροανάπτυξη των παιδιών στην ηλικία των 4 ετών.• Να μελετήσουμε τη σχέση της έκθεσης σε ένα ή και περισσότερους Polyomaviruses και Herpesviruses εως την ηλικία των 4 ετών με την παχυσαρκία και καρδιαγγειακούς δείκτες στα 4 και 6 έτη ζωής.Μεθοδολογία: Ο πληθυσμός της παρούσας διατριβής προέρχεται από τη μελέτη Μητέρας Παιδιού Κρήτης, Μελέτη ΡΕΑ (www.rhea.gr) και περιλαμβάνει ένα δείγμα περίπου 1,000 εγκύων γυναικών (Ελληνίδων και αλλοδαπών) και των παιδιών τους για τις οποίες ο τοκετός πραγματοποιήθηκε σε ένα από τα νοσοκομεία του νομού Ηρακλείου (δημόσια & ιδιωτικά) κατά τη χρονική περίοδο ενός έτους (Μάρτιος 2007-Φεβρουάριος 2008). Οι γυναίκες αυτές προσεγγίστηκαν τη 12η -14η και την 28η -32η εβδομάδα κύησης και τον τοκετό και ακολούθησε παρακολούθηση των παιδιών με συνάντηση στους 6 μήνες ζωής, στα 4 χρόνια και στα 6 χρόνια. Σε ένα τυχαίο δείγμα του πληθυσμού μας έγινε συνάντηση και στα 3 χρόνια ζωής για τους σκοπούς της παρούσας ορολογικής μελέτης.Δείγματα αίματος που συλλέχθηκαν στη γέννηση (ομφαλικό αίμα) (n=626), 3 χρόνια (n=81) και 4 χρόνια (n=690) και είχαν αποθηκευθεί στους -80˚C, στάλθηκαν για ορολογική ανάλυση στο εργαστήριο, Molecular Diagnostics of Oncogenic Infections Department, Infection and Cancer Program, German Cancer Research Center (DKFZ) της Γερμανίας. Η IgG οροαντιδαστικότητα που μετρήθηκε αφορούσε το αντιγόνο VP1 για τους 12 Polyomaviruses (BKPyV, JCPyV, KIPyV, WUPyV, HPyV6, HPyV7, TSPyV, MCPyV, HPyV9 and HPyV10), τέσσερα EBV αντιγόνα (ZEBRA, EBNA-1, EA-D, VCA p18), πέντε CMV αντιγόνα (pp52, pp65, pp150, pp28, CM2), ένα HSV-1 αντιγόνο (gG), ένα HSV-2 αντιγόνο (mgGunique), τέσσερα HHV-8 αντιγόνα (LANA, V-CYCLIN, K8.1, ORF-65) και δώδεκα Helicobacter pylori αντιγόνα (GroEL, UreA, HP0231, NapA, HP0305, HpaA, CagA, HyuA, catalase, VacA, HcpC, Omp). Σημειώνεται ότι η οροαντιδραστικότητα στα δείγματα ομφαλικού αίματος αντιπροσωπεύει την οροαντιδραστικότητα των μητέρων στο τέλος της κύησης. Η οροθετικότητα για κάθε αντιγόνο ορίστηκε χρησιμοποιώντας συγκεκριμένα όρια που στην περίπτωση των Polyomavi

Metabolic profile in early pregnancy is associated with offspring adiposity at 4 years of age: the rhea pregnancy cohort crete, Greece.

CONTEXT: Maternal pre-pregnancy obesity may increase the risk of childhood obesity but it is unknown whether other metabolic factors in early pregnancy such as lipid profile and hypertension are associated with offspring cardiometabolic traits. OBJECTIVE: Our objective was to investigate whether fasting lipid, glucose, and insulin levels during early pregnancy and maternal pre-pregnancy weight status, are associated with offspring adiposity measures, lipid levels and blood pressure at preschool age. DESIGN AND METHODS: The study included 618 mother-child pairs of the pregnancy cohort "Rhea" study in Crete, Greece. Pregnant women were recruited at the first prenatal visit (mean: 12weeks, SD: 0.7). A subset of 348 women provided fasting serum samples for glucose and lipid measurements. Outcomes measures were body mass index, abdominal circumference, sum of skinfold thickness, and blood pressure measurements at 4 years of age. A subsample of 525 children provided non-fasting blood samples for lipid measurements. RESULTS: Pre-pregnancy overweight/obesity was associated with greater risk of offspring overweight/obesity (RR: 1.83, 95%CI: 1.19, 2.81), central adiposity (RR: 1.97, 95%CI: 1.11, 3.49), and greater fat mass by 5.10mm (95%CI: 2.49, 7.71) at 4 years of age. These associations were more pronounced in girls. An increase of 40mg/dl in fasting serum cholesterol levels in early pregnancy was associated with greater skinfold thickness by 3.30mm (95%CI: 1.41, 5.20) at 4 years of age after adjusting for pre-pregnancy BMI and several other confounders. An increase of 10mmHg in diastolic blood pressure in early pregnancy was associated with increased risk of offspring overweight/obesity (RR: 1.22, 95%CI: 1.03, 1.45), and greater skinfold thickness by 1.71mm (95% CI: 0.57, 2.86) at 4 years of age./nCONCLUSIONS: Metabolic dysregulation in early pregnancy may increase the risk of obesity at preschool age.Rhea project was financially supported by European projects (EU FP6-2003-Food-3- NewGeneris, EU FP6. STREP Hiwate, EU FP7Rhea project was financially supported byEuropean projects (EU FP6-2003-Food-3-NewGeneris, EU FP6. STREP Hiwate, EU FP7. ENV.2007.1.2.2.2. Project No 211250 Escape, EUFP7-2008-ENV-1.2.1.4 Envirogenomarkers, EU FP7-HEALTH-2009- single stage CHICOS, EU FP7ENV.2008.1.2.1.6. Proposal No 226285 ENRIECO,EU- FP7- HEALTH-2012 Proposal No 308333HELIX) and the Greek Ministry of Health (Program ofPrevention of obesity and neurodevelopmental disorders in preschool children, in Heraklion district,Crete, Greece: 2011-2014;“Rhea Plus”: Primary Prevention Program of Environmental Risk Factors for Reproductive Health, and Child Health: 2012-15

Gestational sleep deprivation is associated with higher offspring body mass index and blood pressure

Study objectives: The objective of this study was to evaluate the association between gestational sleep deprivation and childhood adiposity and cardiometabolic profile. Methods: Data were used from two population-based birth cohorts (Rhea study and Amsterdam Born Children and their Development study). A total of 3,608 pregnant women and their children were followed up until the age of 11 years. Gestational sleep deprivation was defined as 6 or fewer hours of sleep per day, reported by questionnaire. The primary outcomes included repeated measures of body mass index (BMI), waist circumference, body fat, serum lipids, systolic and diastolic blood pressure (DBP) levels in childhood. We performed a pooled analysis with adjusted linear mixed effect and Cox proportional hazards models. We tested for mediation by birthweight, gestational age, and gestational diabetes. Results: Gestational sleep deprivation was associated with higher BMI (beta; 95% CI: 0.7; 0.4, 1.0 kg/m2) and waist circumference (beta; 95% CI: 0.9; 0.1, 1.6 cm) in childhood, and increased risk for overweight or obesity (HR; 95% CI: 1.4; 1.1, 2.0). Gestational sleep deprivation was also associated with higher offspring DBP (beta; 95% CI: 1.6; 0.5, 2.7 mmHg). The observed associations were modified by sex (all p-values for interaction < 0.05); and were more pronounced in girls. Gestational diabetes and shorter gestational age partly mediated the seen associations. Conclusions: This is the first study showing that gestational sleep deprivation may increase offspring's adiposity and blood pressure, while exploring possible mechanisms. Attention to glucose metabolism and preterm birth might be extra warranted in mothers with gestational sleep deprivation.We are thankful to all the mothers, fathers, and children who participated in the Rhea cohort and Amsterdam Born Children and their Development cohort. The ABCD study has been supported by grants from The Netherlands Organisation for Health Research and Development (ZonMW) and The Netherlands Heart Foundation. Research time of MH was supported by the municipal Amsterdam Healthy Weight Program (Amsterdamse Aanpak Gezond Gewicht). The Rhea project was financially supported by European projects (EU FP6-2003-Food-3-NewGeneris, EU FP6. STREP Hiwate, EU FP7 ENV.2007.1.2.2.2. Project No 211250 Escape, EU FP7-2008-ENV-1.2.1.4 Envirogenomarkers, EU FP7-HEALTH-2009-single stage CHICOS, EU FP7 ENV.2008.1.2.1.6. Proposal No 226285 ENRIECO, EUFP7-HEALTH-2012 Proposal No 308333 HELIX, FP7 European Union project, No. 264357 MeDALL), and the Ministry of health and social solidarity, Greece (Program of Prevention of obesity and neurodevelopmental disorders in preschool children, in Heraklion district, Crete, Greece: 2011–2014; Rhea Plus: Primary Prevention Program of Environmental Risk Factors for Reproductive Health, and Child Health: 2012–15). Dr Lida Chatzi was supported by the National Institute of Environmental Health Sciences (NIEHS/National Institutes of Health (NIH)) grants: R21ES029681, R01ES030691, R01ES029944, R01 ES030364, R21ES028903, and P30ES007048, and by NIH (UH3OD023287). The study was also supported by Seventh Framework Programme and Hartstichting

Influenza vaccination coverage rates and determinants in Greek children until the age of ten (2008-2019), the Rhea mother-child cohort

Background: In Greece, influenza vaccination is currently recommended for children with high-risk conditions. There are limited data on influenza vaccination uptake among Greek children with and without high-risk conditions. We aim to describe the annual influenza vaccination uptake until the age of ten in a population-based mother-child cohort and identify the factors influencing vaccination rates. Methods: Immunization data from the child's health cards at 4 and 10 years were available for 830 and 298 children participating in the Rhea cohort (2008-2019). We calculated vaccination coverage by age, winter season and among children with asthma and obesity for whom the vaccine is indicated. Univariable and multivariable stepwise logistic regression models were utilized to identify the association between several sociodemographic, lifestyle and health-related variables and vaccine uptake by age four. Results: By the ages of four and ten, 37% and 40% of the children, respectively, had received at least one influenza vaccination. Only 2% of the children were vaccinated for all winter seasons during their first four years of life. The vaccination rate was highest at the age of two and during the 2009-2010 season. Vaccination rates for children with asthma and obesity were 18.2% and 13.3% at age four and 8.3% and 2.9% at age ten. About 10% of all vaccines were administered after December and 24% of the children received only one dose upon initial vaccination. Children with younger siblings and those who had experienced more respiratory infections were more likely to be vaccinated by the age of four, while children exposed to smoking were less likely to be vaccinated. Conclusions: Children in our study were more likely to be vaccinated against influenza at an early age with the peak occurring at the age of two. Nonetheless, annual vaccination uptake was uncommon. Vaccination rates of children with asthma and obesity were well below the national target of 75% for individuals with chronic conditions. Certain groups may merit increased attention in future vaccination campaigns such as children raised in families with unfavourable health behaviours.This work was supported by the Greek Ministry of Health (program of prevention of obesity and neurodevelopmental disorders in preschool children, in the Heraklion district, Crete, Greece, 2011–2014; and “Rhea Plus”: Primary Prevention Program of Environmental Risk Factors for Reproductive Health and Child Health, 2012–2015). We acknowledge support from the grant CEX2018-000806-S funded by MCIN/AEI/10.13039/501100011033, and support from the Generalitat de Catalunya through the CERCA Program. The funding sources had no role in the design and conduct of the study; collection, management, analysis, and interpretation of the data; preparation, review, or approval of the manuscript; or the decision to submit the manuscript for publication

Association of trimester-specific gestational weight gain with fetal growth, offspring obesity, and cardiometabolic traits in early childhood

OBJECTIVE: The purpose of this study was to investigate the association of trimester-specific gestational weight gain with offspring fetal growth, obesity risk, and cardiometabolic health outcomes from birth to 4 years of age. STUDY DESIGN: We conducted the present study with 977 motherchild pairs of the pregnancy cohort “Rhea” study in Crete, Greece. We measured birthweight, body mass index from 6 months to 4 years of age, waist circumference, skinfold thickness, blood pressure, and blood levels of lipids, C-reactive protein, and adipose tissue hormones at 4 years of age. We used multiple linear and log Poisson regression models to examine the association of exposure with continuous or binary outcomes, respectively. RESULTS: Greater rate of gestational weight gain in the first trimester of pregnancy (per 200 g/wk) was associated with increased risk of overweight/obesity from 2 years (relative risk [RR], 1.25; 95% confidence interval [CI], 1.09-1.42) to 4 years of age (RR, 1.15; 95% CI, 1.05-1.25), but not with birth size. Each 200 g/wk of weight gain in the first trimester of pregnancy was also associated with greater risk of high waist circumference (RR, 1.13; 95% CI, 1.04-1.23), high sum of skinfold thickness (RR, 1.15; 95% CI, 1.02-1.29), and higher diastolic blood pressure at 4 years of age (beta, 0.43 mm Hg; 95% CI, 0.00-0.86). Greater rate of gestational weight gain during the second and third trimesters of pregnancy (per 200 g/wk) was associated with greater risk of largefor- gestational-age neonates (RR, 1.22; 95% CI, 1.02, 1.45) and higher levels of cord blood leptin (ratio of geometric means, 1.08; 95% CI, 1.00-1.17), but not with child anthropometry at later ages. CONCLUSION: Timing of gestational weight gain may influence childhood cardiometabolic outcomes differentially

Prenatal metal mixtures and child blood pressure in the Rhea mother-child cohort in Greece

Background: Child blood pressure (BP) is predictive of future cardiovascular risk. Prenatal exposure to metals has been associated with higher BP in childhood, but most studies have evaluated elements individually and measured BP at a single time point. We investigated impacts of prenatal metal mixture exposures on longitudinal changes in BP during childhood and elevated BP at 11 years of age. Methods: The current study included 176 mother-child pairs from the Rhea Study in Heraklion, Greece and focused on eight elements (antimony, arsenic, cadmium, cobalt, lead, magnesium, molybdenum, selenium) measured in maternal urine samples collected during pregnancy (median gestational age at collection: 12 weeks). BP was measured at approximately 4, 6, and 11 years of age. Covariate-adjusted Bayesian Varying Coefficient Kernel Machine Regression and Bayesian Kernel Machine Regression (BKMR) were used to evaluate metal mixture impacts on baseline and longitudinal changes in BP (from ages 4 to 11) and the development of elevated BP at age 11, respectively. BKMR results were compared using static versus percentile-based cutoffs to define elevated BP. Results: Molybdenum and lead were the mixture components most consistently associated with BP. J-shaped relationships were observed between molybdenum and both systolic and diastolic BP at age 4. Similar associations were identified for both molybdenum and lead in relation to elevated BP at age 11. For molybdenum concentrations above the inflection points (~ 40-80 μg/L), positive associations with BP at age 4 were stronger at high levels of lead. Lead was positively associated with BP measures at age 4, but only at high levels of molybdenum. Potential interactions between molybdenum and lead were also identified for BP at age 11, but were sensitive to the cutoffs used to define elevated BP. Conclusions: Prenatal exposure to high levels of molybdenum and lead, particularly in combination, may contribute to higher BP at age 4. These early effects appear to persist throughout childhood, contributing to elevated BP in adolescence. Future studies are needed to identify the major sources of molybdenum and lead in this population.This work was supported by NIEHS [grant numbers R21ES029681 (Chatzi, Conti, McConnell), R01ES030691 (Chatzi, Conti, McConnell, Eckel), R01ES029944 (Chatzi, Conti, Margetaki), R01 ES030364 (Chatzi, Conti, McConnell, Eckel), R21ES028903 (Chatzi, McConnell, Eckel), P30ES007048 (Chatzi, Conti, McConnell, Eckel), K99 ES030400 (Howe), R00ES024144 (Farzan), and Horizon 2020 – European Framework Programme for Research and Innovation, ATHLETE, (Chatzi, Conti), P01CA196569, R01CA140561, R01ES016813 (Conti)]. The Rhea project was financially supported by the European Union [grant numbers EU FP6–2003-Food-3-NewGeneris, EU FP6. STREP Hiwate, EU FP7 ENV.2007.1.2.2.2. Project No 211250 Escape, EU FP7–2008-ENV-1.2.1.4 Envirogenomarkers, EU FP7-HEALTH-2009- single stage CHICOS, EU FP7 ENV.2008.1.2.1.6. Proposal No 226285 ENRIECO, EU- FP7- HEALTH-2012 Proposal No 308333 HELIX) and the Greek Ministry of Health

Infection induced SARS-CoV-2 seroprevalence and heterogeneity of antibody responses in a general population cohort study in Catalonia Spain

Sparse data exist on the complex natural immunity to SARS-CoV-2 at the population level. We applied a well-validated multiplex serology test in 5000 participants of a general population study in Catalonia in blood samples collected from end June to mid November 2020. Based on responses to fifteen isotype-antigen combinations, we detected a seroprevalence of 18.1% in adults (n = 4740), and modeled extrapolation to the general population of Catalonia indicated a 15.3% seroprevalence. Antibodies persisted up to 9 months after infection. Immune profiling of infected individuals revealed that with increasing severity of infection (asymptomatic, 1-3 symptoms, ≥ 4 symptoms, admitted to hospital/ICU), seroresponses were more robust and rich with a shift towards IgG over IgA and anti-spike over anti-nucleocapsid responses. Among seropositive participants, lower antibody levels were observed for those ≥ 60 years vs < 60 years old and smokers vs non-smokers. Overweight/obese participants vs normal weight had higher antibody levels. Adolescents (13-15 years old) (n = 260) showed a seroprevalence of 11.5%, were less likely to be tested seropositive compared to their parents and had dominant anti-spike rather than anti-nucleocapsid IgG responses. Our study provides an unbiased estimate of SARS-CoV-2 seroprevalence in Catalonia and new evidence on the durability and heterogeneity of post-infection immunity