Healthy pregnancy requires a tightly regulated materno-fetal dialogue for processes such as embryo implantation, endometrial decidualisation (in the mouse), placentation and maternal adaptation to occur. Disruption of placental development as well as maternal adaptation can lead to fetal intrauterine growth restriction (IUGR) which increases the risk of late miscarriage/stillbirth (e.g. 53% of preterm stillbirth and 26% of term stillbirth are found to be IUGR). Furthermore, IUGR is a risk factor for neurodevelopmental conditions in childhood and for a spectrum of related adult health disorders such as cardiovascular disease and type II diabetes, often termed metabolic syndrome. Despite these pregnancy disorders being common (e.g. 1 in 200 pregnancies results in stillbirth in the UK) the molecular lesion(s) underlying their pathophysiology are poorly understood and in particular those with placental and/or maternal aetiologies most frequently remain unexplained. Here we investigate the hypothesis that poly(A)-binding protein 4 (PABP4) is required for healthy pregnancy in mice. PABP4 is an RNA-binding protein and a member of the PABP family which are central regulators of mRNA translation and stability. Using all four permutations of wild-type and knock-out crosses, we find that maternal PABP4-deficiency results in a reduced litter size and IUGR. The number of implantations at e8.5 were not reduced in Pabp4-/- females, implying that the reduced litter size was not a consequence of decreased ovulation, fertilisation or implantation frequency. Further longitudinal analysis (at e13.5, e15.5 and e18.5) reveals that fetal death primarily occurred between e18.5 and birth, suggesting these mice may provide a unique opportunity to inform on the maternal causes of stillbirth. The onset of IUGR, which was found to be symmetrical in nature, was established by e15.5 preceding the majority of fetal death. During pregnancy, a materno-fetal dialogue directs and responds to changes in gene expression to give rise to the placenta and adapt the maternal physiology. Defects in these processes may result in reduced growth and/or fetal death and were examined in Pabp4-/- mice to shed light on the mechanistic basis of these related phenotypes. Fetal to placental (F:P) weight ratio, whose changes can be indicative of placental insufficiency or placental adaptation in an attempt to aid fetal growth, was found to be increased in Pabp4-/- dams at e15.5 and e18.5 due to the presence of IUGR fetuses with placentas of normal weight. Consistent with this observation, placental volume was unchanged at e18.5. Total placental weight and volume alone fails to discriminate potential differences in the individual placental zones which include the labyrinth zone, where materno-fetal gas and nutrient exchange occur; the junctional zone, which has endocrine functions including those that promote maternal adaptation; and the decidua basalis, derived from the maternal endometrium and is the site of trophoblast invasion and maternal vascular remodelling in early pregnancy. Therefore, volumetric analysis of these zones and the maternal blood spaces, which transcend the decidua basalis and junctional zone, was undertaken. This showed no change in the maternal blood spaces or the labyrinth, the latter being the zone whose size is most frequently altered in IUGR. Critically however, the size of the maternally-derived decidua basalis was increased with a concurrent decrease in the size of the junctional zone. These morphological changes may play a causative role either through directly affecting placental function and/or by the reduced junctional zone failing to promote appropriate maternal adaptation. Alternatively, they may reflect compensatory adaptations to a primary defect elsewhere in the mother. Complementing these morphological studies, functional studies were undertaken: remodelling of maternal vasculature and the resistance index of vessels delivering blood to the fetus were assessed; as was delivery of nutrients to the fetus (measured by fetal glucose); and systemic maternal adaptations (maternal hormonal profile, circulating glucose levels and organ weights). Uterine, umbilical and decidual spiral arteries were examined, but displayed no apparent differences suggestive of normal blood supply to the fetus. However fetal blood glucose was reduced suggesting a reduced delivery of nutrients important for fetal growth. This was not due to lower circulating maternal blood glucose levels, and mRNA levels of the placental glucose transporters Glut-1 and Glut-3 were not reduced but upregulated, suggestive of an attempt to compensate for reduced fetal glucose. Furthermore, upregulation of at least one system A amino acid transporter mRNA, Snat-2, was observed. The maternal physiological state of PABP4-deficient dams showed deviations in some organ weights (e.g. spleen weight is reduced at e13.5 and e15.5) and the levels of some circulating hormones (e.g. estradiol is deceased whereas progesterone is increased at e18.5). However, future work will be required to determine which, if any, of these changes are primary defects rather than downstream consequences and to identify which mis-regulated mRNAs/pathways within in the materno-fetal dialogue underlie the phenotype. Taken together, my results suggest that the regulation of mRNA translation/stability by PABP4 is critical to achieving the correct pattern of gene expression within the materno-fetal dialogue to enable appropriate placentation and maternal adaptation. Furthermore, my results suggest that Pabp4-/- mice provide a unique opportunity to further understand the maternal causes of a spectrum of related pregnancy complications including IUGR, late miscarriage and stillbirth.

The primary objective of antenatal assessment and monitoring is to ensure wellbeing of the fetus and the mother. There are different methods of assessment during pregnancy and in labour. Doppler ultrasound is one of the tests widely used in clinical practice in the evaluation of pregnancies that are at a greater risk of developing maternal or fetal complications due to uteroplacental insufficiency. Doppler ultrasound enables evaluation of sequential changes in circulatory haemodynamics in the fetus by evaluation of the fetus for signs of brain sparing and severity of redistribution of circulation. Recognition of abnormal Doppler flow patterns helps the clinician to optimise the appropriate timing of delivery. Identification of the ‘high risk’ fetus, before any changes of fetal compromise become evident, still remains one of the major dilemmas in contemporary clinical practice. This thesis seeks to explore the role of Doppler monitoring fetal intrauterine central shunts as a method of identifying the ‘high-risk’ fetus before any other established parameters, such as, fetal biometry, fetal weight or flow waveforms in umbilical artery become abnormal. This thesis also evaluates the role of serial Doppler monitoring of fetal central shunts in those fetuses where IUGR has been established. This is based on the premise that the intrauterine shunts are present in fetal circulation to work closely with the placenta to ensure appropriate nutrition and oxygenation of the fetus, bypassing the lungs. Four prospective longitudinal studies were designed to evaluate the role of fetal intrauterine shunts in adaptive response mechanisms in cardiovascular stress. Two models were taken into consideration: an ‘acute cardiovascular stress’ model and a ‘chronic cardiovascular stress’ model. To study the ‘response to acute cardiovascular stress’ in high-risk fetuses, a cohort of mothers undergoing fetal intrauterine transfusion for fetal anaemia were selected. These fetuses were scanned immediately before and after transfusion, and Doppler flows through all the intrauterine shunts were documented and compared with fetoplacental and cerebral circulation. To study the ‘response to chronic cardiovascular stress’, a prospective longitudinal observational study was designed and the sequence of changes in Doppler ultrasound of the fetal central shunts studied and compared with the Doppler flow waveforms of normal pregnancies with a group of pregnancies complicated by uteroplacental insufficiency. Normograms were designed for all the Doppler parameters and flows from adverse pregnancy outcomes were compared to the normogram. The pregnancy outcomes in the longitudinal study were correlated with placental pathology. Our study showed that although changes were demonstrated in the flow patterns within central shunts, these changes were not statistically significant in the ‘acute cardiovascular stress model’, suggesting that there may be other haemodynamic alterations in acute cardiovascular stress. However, in the ‘chronic cardiovascular stress model’, the results suggest that the intrauterine cardiac shunts may play an important role in redistribution of fetal flows in early stages of growth restriction, suggesting that Doppler ultrasound monitoring of foramen ovale can be potentially used as a screening tool to identify high-risk fetuses as early as 16 weeks. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1349883 / Thesis (Ph.D.) - University of Adelaide, School of Paediatrics and Reproductive Health, 2009

Pre-eclampsia and intrauterine growth restriction (IUGR) are common complications of pregnancy with significant implications for mother and fetus. The exact cause is unknown, but current theories suggest that abnormal trophoblast invasion leads to placental under perfusion and hypoxia with the generation of reactive oxygen species (ROS). A result of this insult is increased trophoblast apoptosis. The cell cycle regulator p53 has been identified in increased amounts in the trophoblast in pre-eclampsia and IUGR. It is possible that modulation of this potent apoptotic signal could improve placental function, potentially offering a therapy for these presently untreatable conditions.In these studies, trophoblast apoptosis was induced with Nutlin-3, a synthetic compound which inhibits the binding of p53 to its inhibitor, Mdm2, therefore acting as a p53 activator. Treatment with Nutlin-3 induced apoptotic cell death in BeWo choriocarcinoma cells, primary trophoblast cells and placental villous explants, suggesting that p53 is intrinsic to trophoblast survival. This increase in apoptosis was associated with an increase in p53, Mdm2, p21 and Puma protein expression in BeWo cells and p21 and Puma mRNA in placental villous explants. No effect was observed in Bax expression. Co-treatment with Nutlin-3 and the p53 inhibitor Pifithrin-α reduced apoptosis to the level of control tissue in BeWo cells, primary trophoblast cells and villous explants, suggesting that reducing p53 activity improves trophoblast function.The use of hypoxia and ROS was associated with an increase in apoptosis and p21 and Puma in placental villous explants. Furthermore, the extent of apoptosis and p21 and Puma expression was reduced by co-treatment with Pifithrin-α, suggesting that the ameliorating effects of Pifithrin-α on trophoblast apoptosis are maintained following an insult from a less specific p53 activating mechanism, further supporting the continued investigation of p53 modulation as a therapeutic manipulation of placental function.Pregnant mice were exposed to Nutlin-3 to ascertain the effect of p53 over-activity on pregnancy outcome. The establishment of a model of placental dysfunction in the mouse would have allowed investigation into the effect of p53 inhibition with Pifithrin-α. Despite clear evidence of p53 modulation in maternal tissues, the decidua, no effect was observed upon pregnancy outcome or upon fetal or placental tissue. These findings suggest that the mouse placenta may not be a good model of p53 over-activity, although the exact mechanism of resistance remains unclear.In conclusion, p53 exaggeration is inducible in trophoblast with Nutlin-3, hypoxia and ROS. Furthermore, Pifithrin-α is able to reduce this effect, suggesting that the p53 pathway may be a realistic target for modulation in the placenta.

Intrauterine growth restriction causes impaired growth and development of mammalian fetus, and leads to long-term negative effect on postnatal growth. Among domestic animals, pigs exhibit the most severe naturally occurring IUGR and reduced postnatal muscle growth. The objectives of this research project were to: 1) determine muscle stem cell characteristics in IUGR pigs; 2) determine how intrauterine growth restriction alters protein deposition in skeletal muscle; 3) investigate whether branched-chain amino acids (BCAA) are able to enhance protein synthesis in intrauterine growth restricted (IUGR) pig muscle. Newborn piglets were considered normal body weight (NBWT) or IUGR when birth weight was within ± 0.5 SD and -2 SD of litter average respectively. Muscle satellite cell numbers, believed to be the major nuclei source for postnatal muscle growth, were lower in newborn IUGR pigs which could result in reduced muscle hypertrophy potential. In addition, cultures derived from IUGR muscle satellite cells had a lower fusion percentage. Fewer satellite cells and impaired differentiation ability may contributor to impaired muscle growth in these pigs. Protein synthesis rate was significantly lower in IUGR pig hindquarter in the first hour after feeding, but BCAA supplementation had no effect on protein synthesis in IUGR pigs. Further, eukaryotic translation initiation factor 4E (eIF4E) expression is down regulated in IUGR pig muscle. These results suggest that impaired translation initiation may provide a plausible explanation for the lower protein synthesis rates observed in IUGR pigs. Overall, reduced muscle stem cell number and changes in their activity, as well as impaired translation initiation may be important explanations for compromised postnatal muscle growth in intrauterine growth restricted pigs. / Ph. D.

Η ενδομήτρια καθυστέρηση ανάπτυξης ορίζεται ως παθολογική μείωση του ρυθμού εμβρυικής ανάπτυξης που καταλήγει σε ένα έμβρυο το οποίο δεν έχει την αναμενόμενη ανάπτυξη σύμφωνα με την ηλικία κύησης και το οποίο διατρέχει κίνδυνο αυξημένης περιγεννητικής νοσηρότητας και θνησιμότητας. Ενδομήτρια καθυστέρηση ανάπτυξης είναι η ανικανότητα του εμβρύου να διατηρήσει τον αναμενόμενο ρυθμό ανάπτυξης ανεξάρτητα αν το βάρος του είναι κάτω από τη 10η εκατοστιαία θέση. Όταν η ανάπτυξη του εμβρύου είναι κάτω από την 5η εκατοστιαία θέση τότε ίσως να μπορεί να θεωρηθεί πραγματική ενδομήτρια καθυστέρηση ανάπτυξης. Σύμφωνα με το αμερικανικό κολέγιο γυναικολόγων και μαιευτήρων American College of Obstetricians and Gynecologists (ACOG) η ενδομήτρια καθυστέρηση ανάπτυξης ορίζεται ως το έμβρυο που αποτυγχάνει να έχει τον αναμενόμενο ρυθμό ανάπτυξης. Πολλές φορές οι όροι ενδομήτρια καθυστέρηση ανάπτυξης (IUGR) και μικρό για την ηλικία κύησης (SGA) συγχέονται και για αυτό το λόγο διευκρινίζεται ότι ο όρος μικρό για την ηλικία κύησης (SGA) αναφέρεται σε έμβρυο του οποίου το εκτιμώμενο βάρος είναι κάτω από τη 10η εκατοστιαία θέση. Από το σύνολο των μικρών για την ηλικία κύησης (SGA) εμβρύων το 40% είναι υγιή, 20% είναι μικρά εξαιτίας χρωμοσωμικών και περιβαλλοντικών παραγόντων και 40% διατρέχουν ιδιαίτερα αυξημένο κίνδυνο ενδομήτριου θανάτου και σχετίζονται με ενδομήτρια καθυστέρηση ανάπτυξης. Η ενδομήτρια καθυστέρηση ανάπτυξης είναι μια πολυσυστηματική διαταραχή και διακρίνεται σε: Α) Συμμετρική. Όταν η ανάπτυξη της κεφαλής και της κοιλιάς επιβραδύνονται συμμετρικά. Είναι λιγότερο συχνή, 20%-30%. Β) Ασύμμετρη. Δυσανάλογη ελάττωση του μεγέθους της κοιλιάς σε σχέση με αυτό της κεφαλής. Είναι περισσότερο συχνή, 70%-80%, και οφείλεται συνήθως σε μητροπλακουντιακή ανεπάρκεια και συμβαίνει μετά την 28η εβδομάδα. Στη φάση αυτή γίνεται η μεγαλύτερη εναπόθεση λίπους. Η ενδομήτρια καθυστέρηση ανάπτυξης αποτελεί κύριο πρόβλημα στην περιγεννητική ιατρική, είναι η ΔΕΥΤΕΡΗ αιτία θανάτου μετά την προωρότητα. 52% των ανεξήγητων ξαφνικών θανάτων των εμβρύων συσχετίζονται με ανάπτυξη IUGR. 10% των περιπτώσεων περιγεννητικής θνησιμότητας στην Ευρώπη είναι αποτέλεσμα μη διαγνωσμένης IUGR. Η επίπτωση της ενδομήτριας καθυστέρησης ανάπτυξης είναι 8% στον γενικό πληθυσμό. / Intrauterine growth restriction (IUGR) is the failure of the fetus to achieve his/her intrinsic growth potential and is associated with significantly increased perinatal morbidity and mortality. IUGR affects 10% of pregnancies and perinatal mortality rates are 4-8 times higher for infants with this disorder. Intrauterine growth restriction is not a specific disease entity with a unique pathophysiology, but the result of suboptimal intrauterine growth conditions in conjunction with a variety of disorders from genetic to metabolic, vascular, coagulative, autoimmune, as well as infectious. Newborns with intrauterine growth restriction (IUGR) present reduced fat mass and undergo adaptational changes of endocrine/metabolic mechanisms as a result of intrauterine malnutrition. Recently, it was shown that fat-secreted adipokines such as visfatin, a novel adipokine improving glucose tolerance through insulin-mimetic effects, and vaspin , profoundly influence insulin sensitivity and energy metabolism.

Medical diagnosis is the process of attempting to determine and/or identify a possible disease or disorder. This process is revealed by biomarkers, defined by The Food and Drug Administration (FDA) as “characteristics that are objectively measured and evaluated as indicators of normal biologic processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention”. The process of biomarker discovery has been boosted in the last years by proteomics, a research discipline that takes a snapshot of the entire wealth of proteins in an organism/ tissue/ cell/ body fluid. An implementation of the analysis methods can help in isolate proteins present in the low range of concentrations, such as biomarkers very often are. An established biomarker can further be measured with the help of biosensors, devices that can be employed in the point-of care diagnostics. This PhD thesis shows and discusses the results of three projects in the field of protein biomarkers discovery and quantification. The first project exploited proteomics techniques to find relevant protein markers for Intrauterine Growth Restriction (IUGR) in cordonal blood serum (UCS) and amniotic fluid (AF). A 14 proteins in UCS and 11 in AF were successfully identified and found to be differentially expressed. Molecularly Imprinted Polymers (MIPs) directed towards proteins and peptides containing phosphotyrosine were then produced, with the final goal of selectively extracting phosphopeptides from a peptide mixture. An alteration of the phosphorylation pattern is in fact often associated to important diseases such as cancer. The polymers were produced as nanoparticles, that were characterised with Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM). A recipe was also tested for binding capacity towards phosphotyrosine. A Surface Plasmon Resonance (SPR) biosensor to quantify hepcidin hormone was finally produced. This is the major subject in iron homeostasis in vertebrates and marker of iron unbalance diseases. A calibration curve was made and affinity/kinetic parameters for the ligand employed were measured.

The thrifty phenotype hypothesis proposes reduced nutrition alters the trajectory of development of metabolic regulatory systems to produce a phenotype better fitted to an environment of decreased later-life nutrient availability. Because organisms have physiological mechanisms for coping with poor nutrition, they may have sociobehavioral mechanisms as well. Aggressive behavior, especially in the context of feeding competition, may be advantageous in such environments. There could be an association between aggression and intrauterine growth restriction (IUGR), which can result from low maternal food intake during pregnancy. The main hypotheses of this study are [1] IUGR offspring demonstrate higher rates of aggressive behavior and [2] IUGR offspring attain higher ranks. Behavioral observations were conducted on 22 juvenile baboons (Papio sp., ages 3-5 yrs) living in groups. Male IUGR (n = 4) and female IUGR (n = 5) were offspring of mothers fed 70% the same feed eaten by control (CTR) mothers in pregnancy and lactation. CTR males (n = 8) and CTR females (n = 5) were offspring of mothers fed ad libitum. Some authorities recommend this moderate level of dietary restriction for health and longevity. Offspring have not experienced dietary restriction since weaning. IUGR, compared to CTR, showed significantly increased rates of aggressive behavior, especially threat displays. Differences were more dramatic in males than in females. IUGR baboons performed the affiliative display behaviors lipsmack and chatter at elevated rates too, perhaps to counteract the effects of increased aggressive displays. IUGR females exhibited increased rates of stereotypical chewing behavior, while IUGR males exhibited decreased rates of play behavior, possibly indicating elevated anxiety levels. There was only limited support for condition-based differences in rank. Elevated rates of aggression in IUGR baboons may reflect an aggressive behavioral phenotype that enhances fitness by improving access to resources. Alternatively, they could be a non-adaptive result of neurodevelopment with a potentially negative impact on fitness. Unraveling the dynamic relationship between experiences and development is essential for understanding how phenotypes are formed. This will improve the ability of mothers to assess benefits of different nutritional strategies, leading to healthier individuals not just during growth and development, but throughout life.

Introdução: Eventos perinatais podem afetar a saúde do indivíduo a médio e em longo prazo. Além disso, a relação mãe-bebê se relaciona com o risco para psicopatologias durante a vida, e também parece influenciar a nutrição e o crescimento da criança. Crianças nascidas com restrição de crescimento intrauterino (RCIU), especialmente as meninas, apresentam maior consumo de alimentos palatáveis em várias fases do desenvolvimento, o que aumenta o risco para obesidade ao longo da vida. Nosso grupo vem demonstrando que alterações no sistema de recompensa do cérebro possam estar envolvidas. O uso da intervenção musical tem demonstrado melhora em uma série de parâmetros comportamentais e fisiológicos, assim como a aceitação alimentar em bebês nascidos com baixo peso no início da vida. Recentemente, estudos de neuroimagem têm sugerido que a exposição à música ativa o sistema de recompensa do cérebro. Entretanto, ainda não se sabe os efeitos de uma intervenção musical no início da vida sobre estes desfechos em longo prazo. Objetivos: Avaliar o impacto de uma intervenção em pares de mães e bebês (exposição a aulas de música) sobre desfechos relacionados à saúde da criança em longo prazo, buscando associálos com o peso ao nascer. Metodologia: Estudo longitudinal controlado que avaliou 56 crianças com idades entre 5 e 9 anos em desfechos antropométricos, nutricionais, comportamentais e de expressão musical. O grupo exposto foi recrutado de uma amostra de crianças que participou de uma intervenção musical estruturada de 2004 a 2007 no Curso de Extensão Música para Bebês do Departamento de Música do Instituto de Artes da UFRGS. O grupo não exposto foi recrutado de uma amostra de controles populacionais da mesma idade, na área de abrangência da Unidade Básica de Saúde Santa Cecília. Uma série de General Linear Model (GLMs) foram feitas, ajustadas para nível sócio-econômico e educação materna, para avaliar a interação entre a exposição à música, o peso ao nascer e sexo sobre o consumo de alimentos através de questionário de frequência alimentar. Resultados: Cinquenta e seis crianças foram avaliadas, sendo 28 expostas. Não houve diferença significativa entre os grupos exposto e não exposto na distribuição do sexo (p=0.42). Não foram encontradas diferenças estatisticamente significativas quanto aos instrumentos Parental Bonding Instrument (PBI), domínio estabilidade familiar no Recursos do Ambiente Familiar (RAF), Questionário sobre o temperamento da criança (CBQ), Questionário sobre o comportamento alimentar da criança (CEBQ), Critério de Classificação Econômica Brasil e Questionário de Frequência Alimentar (QFA). Tampouco na avaliação musical ou nas medidas de cortisol. Entretanto, quanto à escolaridade do responsável, ao nível sócioeconômico e ao domínio processos proximais do RAF, os dados apresentaram diferenças estatisticamente significativas (p<0,001). Houve diferença marginal quanto à Razão de Crescimento Fetal (p=0,051), à idade das crianças (p=0,003) e ao domínio ligação famíliaescola do RAF (p=0,041). Há uma interação entre o peso ao nascer, sexo e exposição à música sobre o consumo de açúcares na infância (Wald=7,87, df=2, p=0.02). A análise da interação mostra que, nas meninas não expostas à música, há aumento do consumo deste alimento conforme o peso ao nascer diminui (B=-8,673, p<0.0001), sem efeito nas expostas (B=3,352, p=0,15) ou nos meninos (expostos B=2,870, p=0.44; não expostos B=3,706, p=0,236). Não foram encontrados efeitos na análise de outros alimentos como frutas ou gorduras, mostrando que o efeito é específico para o doce. Conclusões: Os dados sugerem que intervenção musical em bebês pode moderar os efeitos da RCIU sobre a preferência a alimentos palatáveis na infância em meninas. Acreditamos que a musicalização de bebês pode ser uma intervenção relevante em populações vulneráveis como a das crianças nascidas com RCIU. / Introduction: Perinatal events can have mid- to long-term effects on a person's health. Besides, the mother-baby relationship is related to the risk of psychopathologies during life, and also seems to influence the nutrition and growth of the child. Children born after intrauterine growth restriction (IUGR), especially girls, present a higher intake of palatable foods in many stages of development, which increases the risk of obesity throughout life. Our group has demonstrated that alterations in the brain's rewarding system may be involved. The use of musical interventions has helped with a series of behavioral and physiological parameters, as well as with the acceptance of food in babies born with low birth weight. Recently, neuroimage studies have suggested that exposition to music activates the brain's rewarding system. However, the effects of a musical intervention in the beginning of life on these parameters are still unknown. Objectives: The objective is to evaluate the impact of an intervention in mother-baby pairs (exposed to music classes) on the outcomes related to the health of the child in the long term, associating it to the birth weight. Methodology: This is a controlled longitudinal study that evaluated 56 children, ages 5 to 9, regarding anthropometric, nutritional, behavioral and musical expression outcomes. The exposed group was recruited from a sample of children who participated in a structured musical intervention which happened from 2004 to 2007 at the Music for Babies Extension Course of the Art Institute of the Federal University of Rio Grande do Sul (UFRGS), Brazil. The nonexposed group was recruited from a populational communitarian age-matched sample, in the area encompassed by the Santa Cecília Public Hospital. A series of General Linear Models (GLMs) were performed, according to SES and maternal education. This was done to evaluate the interaction between exposure to music, birth weight and sex about the consumption of food through a food frequency questionnaire. Results: Fifty-six children were evaluated, and 28 were exposed. There was no significant difference between the exposed and the nonexposed group regarding sex (p=0.42). Statistically significant differences were not found regarding the instruments Parental Bonding Instrument (PBI), family stability with Resources of Family Environment (RAF - Recursos de Ambiente Familiar), Children's Behavior Questionnaire (CBQ), Children's Eating Behavior Questionnaire (CEBQ), Brazil's Socioeconomic Classification Criteria, and Food Frequency Questionnaire. There was no significant difference in the musical or the cortisol evaluation. However, regarding the education of the mother, the socioeconomic status, and the grasp of closeness processes with RAF, the data presented statistically significant differences (p<0,001). There was a marginal difference regarding the Fetal Growth Ratio (p=0.051), the children's age (p=0.003) and the connection between family-school evaluated in RAF (p=0.041). There is an interaction between birth weight and exposure to music on the intake of sugar during childhood (Wald=7.87, df=2, p=0.02). The analysis of the interaction shows that girls who were not exposed to music had an increased consumption of this food as the birth weight lowers (B=- 8.673, p<0.0001). There was no effect on the girls who were exposed (B=3.352, p=0.15) nor on the boys (exposed B=2.870, p=0.44; nonexposed B=3.706, p=0.236). This interaction was not found regarding other foods, such as fruits or fats, which shows this is specific to sweets. Conclusions: The data suggest that musical intervention for babies can control the effects of IUGR about the preferences for palatable foods during childhood in girls. We believe that the musicalization of babies can be a relevant intervention in vulnerable populations, such as children born after IUGR.

Introdução: Eventos perinatais podem afetar a saúde do indivíduo a médio e em longo prazo. Além disso, a relação mãe-bebê se relaciona com o risco para psicopatologias durante a vida, e também parece influenciar a nutrição e o crescimento da criança. Crianças nascidas com restrição de crescimento intrauterino (RCIU), especialmente as meninas, apresentam maior consumo de alimentos palatáveis em várias fases do desenvolvimento, o que aumenta o risco para obesidade ao longo da vida. Nosso grupo vem demonstrando que alterações no sistema de recompensa do cérebro possam estar envolvidas. O uso da intervenção musical tem demonstrado melhora em uma série de parâmetros comportamentais e fisiológicos, assim como a aceitação alimentar em bebês nascidos com baixo peso no início da vida. Recentemente, estudos de neuroimagem têm sugerido que a exposição à música ativa o sistema de recompensa do cérebro. Entretanto, ainda não se sabe os efeitos de uma intervenção musical no início da vida sobre estes desfechos em longo prazo. Objetivos: Avaliar o impacto de uma intervenção em pares de mães e bebês (exposição a aulas de música) sobre desfechos relacionados à saúde da criança em longo prazo, buscando associálos com o peso ao nascer. Metodologia: Estudo longitudinal controlado que avaliou 56 crianças com idades entre 5 e 9 anos em desfechos antropométricos, nutricionais, comportamentais e de expressão musical. O grupo exposto foi recrutado de uma amostra de crianças que participou de uma intervenção musical estruturada de 2004 a 2007 no Curso de Extensão Música para Bebês do Departamento de Música do Instituto de Artes da UFRGS. O grupo não exposto foi recrutado de uma amostra de controles populacionais da mesma idade, na área de abrangência da Unidade Básica de Saúde Santa Cecília. Uma série de General Linear Model (GLMs) foram feitas, ajustadas para nível sócio-econômico e educação materna, para avaliar a interação entre a exposição à música, o peso ao nascer e sexo sobre o consumo de alimentos através de questionário de frequência alimentar. Resultados: Cinquenta e seis crianças foram avaliadas, sendo 28 expostas. Não houve diferença significativa entre os grupos exposto e não exposto na distribuição do sexo (p=0.42). Não foram encontradas diferenças estatisticamente significativas quanto aos instrumentos Parental Bonding Instrument (PBI), domínio estabilidade familiar no Recursos do Ambiente Familiar (RAF), Questionário sobre o temperamento da criança (CBQ), Questionário sobre o comportamento alimentar da criança (CEBQ), Critério de Classificação Econômica Brasil e Questionário de Frequência Alimentar (QFA). Tampouco na avaliação musical ou nas medidas de cortisol. Entretanto, quanto à escolaridade do responsável, ao nível sócioeconômico e ao domínio processos proximais do RAF, os dados apresentaram diferenças estatisticamente significativas (p<0,001). Houve diferença marginal quanto à Razão de Crescimento Fetal (p=0,051), à idade das crianças (p=0,003) e ao domínio ligação famíliaescola do RAF (p=0,041). Há uma interação entre o peso ao nascer, sexo e exposição à música sobre o consumo de açúcares na infância (Wald=7,87, df=2, p=0.02). A análise da interação mostra que, nas meninas não expostas à música, há aumento do consumo deste alimento conforme o peso ao nascer diminui (B=-8,673, p<0.0001), sem efeito nas expostas (B=3,352, p=0,15) ou nos meninos (expostos B=2,870, p=0.44; não expostos B=3,706, p=0,236). Não foram encontrados efeitos na análise de outros alimentos como frutas ou gorduras, mostrando que o efeito é específico para o doce. Conclusões: Os dados sugerem que intervenção musical em bebês pode moderar os efeitos da RCIU sobre a preferência a alimentos palatáveis na infância em meninas. Acreditamos que a musicalização de bebês pode ser uma intervenção relevante em populações vulneráveis como a das crianças nascidas com RCIU. / Introduction: Perinatal events can have mid- to long-term effects on a person's health. Besides, the mother-baby relationship is related to the risk of psychopathologies during life, and also seems to influence the nutrition and growth of the child. Children born after intrauterine growth restriction (IUGR), especially girls, present a higher intake of palatable foods in many stages of development, which increases the risk of obesity throughout life. Our group has demonstrated that alterations in the brain's rewarding system may be involved. The use of musical interventions has helped with a series of behavioral and physiological parameters, as well as with the acceptance of food in babies born with low birth weight. Recently, neuroimage studies have suggested that exposition to music activates the brain's rewarding system. However, the effects of a musical intervention in the beginning of life on these parameters are still unknown. Objectives: The objective is to evaluate the impact of an intervention in mother-baby pairs (exposed to music classes) on the outcomes related to the health of the child in the long term, associating it to the birth weight. Methodology: This is a controlled longitudinal study that evaluated 56 children, ages 5 to 9, regarding anthropometric, nutritional, behavioral and musical expression outcomes. The exposed group was recruited from a sample of children who participated in a structured musical intervention which happened from 2004 to 2007 at the Music for Babies Extension Course of the Art Institute of the Federal University of Rio Grande do Sul (UFRGS), Brazil. The nonexposed group was recruited from a populational communitarian age-matched sample, in the area encompassed by the Santa Cecília Public Hospital. A series of General Linear Models (GLMs) were performed, according to SES and maternal education. This was done to evaluate the interaction between exposure to music, birth weight and sex about the consumption of food through a food frequency questionnaire. Results: Fifty-six children were evaluated, and 28 were exposed. There was no significant difference between the exposed and the nonexposed group regarding sex (p=0.42). Statistically significant differences were not found regarding the instruments Parental Bonding Instrument (PBI), family stability with Resources of Family Environment (RAF - Recursos de Ambiente Familiar), Children's Behavior Questionnaire (CBQ), Children's Eating Behavior Questionnaire (CEBQ), Brazil's Socioeconomic Classification Criteria, and Food Frequency Questionnaire. There was no significant difference in the musical or the cortisol evaluation. However, regarding the education of the mother, the socioeconomic status, and the grasp of closeness processes with RAF, the data presented statistically significant differences (p<0,001). There was a marginal difference regarding the Fetal Growth Ratio (p=0.051), the children's age (p=0.003) and the connection between family-school evaluated in RAF (p=0.041). There is an interaction between birth weight and exposure to music on the intake of sugar during childhood (Wald=7.87, df=2, p=0.02). The analysis of the interaction shows that girls who were not exposed to music had an increased consumption of this food as the birth weight lowers (B=- 8.673, p<0.0001). There was no effect on the girls who were exposed (B=3.352, p=0.15) nor on the boys (exposed B=2.870, p=0.44; nonexposed B=3.706, p=0.236). This interaction was not found regarding other foods, such as fruits or fats, which shows this is specific to sweets. Conclusions: The data suggest that musical intervention for babies can control the effects of IUGR about the preferences for palatable foods during childhood in girls. We believe that the musicalization of babies can be a relevant intervention in vulnerable populations, such as children born after IUGR.

Introdução: Eventos perinatais podem afetar a saúde do indivíduo a médio e em longo prazo. Além disso, a relação mãe-bebê se relaciona com o risco para psicopatologias durante a vida, e também parece influenciar a nutrição e o crescimento da criança. Crianças nascidas com restrição de crescimento intrauterino (RCIU), especialmente as meninas, apresentam maior consumo de alimentos palatáveis em várias fases do desenvolvimento, o que aumenta o risco para obesidade ao longo da vida. Nosso grupo vem demonstrando que alterações no sistema de recompensa do cérebro possam estar envolvidas. O uso da intervenção musical tem demonstrado melhora em uma série de parâmetros comportamentais e fisiológicos, assim como a aceitação alimentar em bebês nascidos com baixo peso no início da vida. Recentemente, estudos de neuroimagem têm sugerido que a exposição à música ativa o sistema de recompensa do cérebro. Entretanto, ainda não se sabe os efeitos de uma intervenção musical no início da vida sobre estes desfechos em longo prazo. Objetivos: Avaliar o impacto de uma intervenção em pares de mães e bebês (exposição a aulas de música) sobre desfechos relacionados à saúde da criança em longo prazo, buscando associálos com o peso ao nascer. Metodologia: Estudo longitudinal controlado que avaliou 56 crianças com idades entre 5 e 9 anos em desfechos antropométricos, nutricionais, comportamentais e de expressão musical. O grupo exposto foi recrutado de uma amostra de crianças que participou de uma intervenção musical estruturada de 2004 a 2007 no Curso de Extensão Música para Bebês do Departamento de Música do Instituto de Artes da UFRGS. O grupo não exposto foi recrutado de uma amostra de controles populacionais da mesma idade, na área de abrangência da Unidade Básica de Saúde Santa Cecília. Uma série de General Linear Model (GLMs) foram feitas, ajustadas para nível sócio-econômico e educação materna, para avaliar a interação entre a exposição à música, o peso ao nascer e sexo sobre o consumo de alimentos através de questionário de frequência alimentar. Resultados: Cinquenta e seis crianças foram avaliadas, sendo 28 expostas. Não houve diferença significativa entre os grupos exposto e não exposto na distribuição do sexo (p=0.42). Não foram encontradas diferenças estatisticamente significativas quanto aos instrumentos Parental Bonding Instrument (PBI), domínio estabilidade familiar no Recursos do Ambiente Familiar (RAF), Questionário sobre o temperamento da criança (CBQ), Questionário sobre o comportamento alimentar da criança (CEBQ), Critério de Classificação Econômica Brasil e Questionário de Frequência Alimentar (QFA). Tampouco na avaliação musical ou nas medidas de cortisol. Entretanto, quanto à escolaridade do responsável, ao nível sócioeconômico e ao domínio processos proximais do RAF, os dados apresentaram diferenças estatisticamente significativas (p<0,001). Houve diferença marginal quanto à Razão de Crescimento Fetal (p=0,051), à idade das crianças (p=0,003) e ao domínio ligação famíliaescola do RAF (p=0,041). Há uma interação entre o peso ao nascer, sexo e exposição à música sobre o consumo de açúcares na infância (Wald=7,87, df=2, p=0.02). A análise da interação mostra que, nas meninas não expostas à música, há aumento do consumo deste alimento conforme o peso ao nascer diminui (B=-8,673, p<0.0001), sem efeito nas expostas (B=3,352, p=0,15) ou nos meninos (expostos B=2,870, p=0.44; não expostos B=3,706, p=0,236). Não foram encontrados efeitos na análise de outros alimentos como frutas ou gorduras, mostrando que o efeito é específico para o doce. Conclusões: Os dados sugerem que intervenção musical em bebês pode moderar os efeitos da RCIU sobre a preferência a alimentos palatáveis na infância em meninas. Acreditamos que a musicalização de bebês pode ser uma intervenção relevante em populações vulneráveis como a das crianças nascidas com RCIU. / Introduction: Perinatal events can have mid- to long-term effects on a person's health. Besides, the mother-baby relationship is related to the risk of psychopathologies during life, and also seems to influence the nutrition and growth of the child. Children born after intrauterine growth restriction (IUGR), especially girls, present a higher intake of palatable foods in many stages of development, which increases the risk of obesity throughout life. Our group has demonstrated that alterations in the brain's rewarding system may be involved. The use of musical interventions has helped with a series of behavioral and physiological parameters, as well as with the acceptance of food in babies born with low birth weight. Recently, neuroimage studies have suggested that exposition to music activates the brain's rewarding system. However, the effects of a musical intervention in the beginning of life on these parameters are still unknown. Objectives: The objective is to evaluate the impact of an intervention in mother-baby pairs (exposed to music classes) on the outcomes related to the health of the child in the long term, associating it to the birth weight. Methodology: This is a controlled longitudinal study that evaluated 56 children, ages 5 to 9, regarding anthropometric, nutritional, behavioral and musical expression outcomes. The exposed group was recruited from a sample of children who participated in a structured musical intervention which happened from 2004 to 2007 at the Music for Babies Extension Course of the Art Institute of the Federal University of Rio Grande do Sul (UFRGS), Brazil. The nonexposed group was recruited from a populational communitarian age-matched sample, in the area encompassed by the Santa Cecília Public Hospital. A series of General Linear Models (GLMs) were performed, according to SES and maternal education. This was done to evaluate the interaction between exposure to music, birth weight and sex about the consumption of food through a food frequency questionnaire. Results: Fifty-six children were evaluated, and 28 were exposed. There was no significant difference between the exposed and the nonexposed group regarding sex (p=0.42). Statistically significant differences were not found regarding the instruments Parental Bonding Instrument (PBI), family stability with Resources of Family Environment (RAF - Recursos de Ambiente Familiar), Children's Behavior Questionnaire (CBQ), Children's Eating Behavior Questionnaire (CEBQ), Brazil's Socioeconomic Classification Criteria, and Food Frequency Questionnaire. There was no significant difference in the musical or the cortisol evaluation. However, regarding the education of the mother, the socioeconomic status, and the grasp of closeness processes with RAF, the data presented statistically significant differences (p<0,001). There was a marginal difference regarding the Fetal Growth Ratio (p=0.051), the children's age (p=0.003) and the connection between family-school evaluated in RAF (p=0.041). There is an interaction between birth weight and exposure to music on the intake of sugar during childhood (Wald=7.87, df=2, p=0.02). The analysis of the interaction shows that girls who were not exposed to music had an increased consumption of this food as the birth weight lowers (B=- 8.673, p<0.0001). There was no effect on the girls who were exposed (B=3.352, p=0.15) nor on the boys (exposed B=2.870, p=0.44; nonexposed B=3.706, p=0.236). This interaction was not found regarding other foods, such as fruits or fats, which shows this is specific to sweets. Conclusions: The data suggest that musical intervention for babies can control the effects of IUGR about the preferences for palatable foods during childhood in girls. We believe that the musicalization of babies can be a relevant intervention in vulnerable populations, such as children born after IUGR.