Preterm milk oligosaccharides during the first month of lactation.

OBJECTIVE: Oligosaccharides represent one of the main components of human milk, and they have been assigned important biological functions for newborns. Qualitatively and quantitatively, their presence in milk is strictly related to the expression of the mother's Se and/or Le genes, on the basis of which 4 different milk groups have been described. The aim of the study was to provide new data on the oligosaccharide composition of preterm milk in relation to the 4 groups. METHODS: High-pH anion-exchange chromatography was used to quantify levels of 23 oligosaccharides and lactose in 252 milk samples collected from 63 mothers during the first month of lactation and to identify the 4 milk groups. RESULTS: Substantial differences in oligosaccharide contents were found within the groups and were strictly related to the presence or absence of specific fucosyl-oligosaccharides. The highest concentration was found in group 1 (>20 g/L), the lowest level was found in group 4 (∼10 g/L), and intermediate values were observed in groups 2 and 3. No statistically significant differences in lactose concentrations were observed among the groups. CONCLUSIONS: Our data confirm lower lactose concentrations in preterm milk, compared with term milk, and they provide the first detailed characterization of oligosaccharides in preterm milk, demonstrating important differences in oligosaccharide contents in the 4 groups. These differences might exert an influence on several biological functions that are particularly important for preterm infants and currently are attributed to milk oligosaccharides.

Effect of two pasteurization methods on the protein content of human milk.

The Holder method is the recommended pasteurization method for human milk banks, as it ensures the microbiological safety of human milk (HM). The loss of some biologically active milk components, due to the heat treatment, is a main limit to the diffusion of donor HM. High-temperature short-time (HTST) pasteurization may be an alternative to maintain the nutritional and immunological quality of HM. The aim of the present study was to compare the impact of Holder and HTST pasteurization on the HM protein profile. The protein patterns of HTST-treated milk and raw milk were similar. The Holder method modified bile salt-stimulated lipase, lactoferrin and components of the immune system. The HTST method preserved the integrity of bile salt-stimulated lipase, lactoferrin and, to some extent, of IgAs. Holder pasteurization decreased the amount of bile salt-stimulated lipase and inactivated the remaining molecules, while the HTST method did not alter its activity. Pasteurization increased the bioavailable lysine quantity. HTST pasteurization seems to better retain the protein profile and some of the key active components of donor HM.

Reference values of blood cell counts in the first days of life.

The lack of updated neonatal reference values for hematological parameters impacts significantly with clinical management of both healthy and sick newborns. The present pilot study was thus aimed at assessing updated hematological Italian reference values in late preterm and term newborns. From January 2004 to December 2008 hematological laboratory tests were performed in 1175 newborns (820 healthy and 355 sick controls) between 33-41 weeks of gestation, during the first four days after birth. Hematological parameters were sorted for gender and gestational age and statistically analyzed. No gender-related differences were observed at different weeks of gestation and no significant differences were found when study population was sub-grouped for late preterm and term newborns. During the first 4 days of life erythrocytes and platelets remained stable whilst white blood cell counts and differentials were significantly modified. This study shares updated reference values for hematological parameters in the early phases after birth and offers additional support for improving the management of sick infants.

Neonatal anthropometric charts: the Italian neonatal study compared with other European studies.

BACKGROUND AND OBJECTIVE: This was a nationwide prospective study carried out in Italy between 2005 and 2007, involving 34 centers with a neonatal intensive care unit. The study reports the Italian Neonatal Study charts for weight, length, and head circumference of singletons born between 23 and 42 gestational weeks, comparing them with previous Italian data and with the most recent data from European countries. PATIENTS AND METHODS: Single live born babies with ultrasound assessment of gestational age within the first trimester, and with both parents of Italian origin. Only fetal hydrops and major congenital anomalies diagnosed at birth were excluded. The reference set consists of 22,087 girls and 23,375 boys. RESULTS: At each gestational age, boys are heavier than girls by about 4%. Later-born neonates are heavier than firstborn neonates by about 3%. The effects of sex and birth order on length and head circumference are milder. No differences were observed between babies born in central-north Italy and southern Italy. A large variability emerged among European neonatal charts, resulting in huge differences in the percentage of Italian Neonatal Study neonates below the 10th centile, which is traditionally used to define small-for-gestational-age babies. In the last 2 decades prominent changes in the distribution of birth weight emerged in Italy and in the rest of Europe, in both term and preterm neonates. CONCLUSIONS: The existing European neonatal charts, based on more or less recent data, were found to be inappropriate for Italy. Until an international standard is developed, the use of national updated reference charts is recommended.

Detailed proteomic analysis on DM: insight into its hypoallergenicity.

Successful therapy in cow milk (CM) protein allergy rests upon completely eliminating CM proteins from the child's diet: it is thus necessary to provide a replacement food. Donkey milk (DM) has recently aroused scientific and clinical interest, above all among paediatric allergologists. A deeper knowledge of proteins in DM is necessary to evaluate the immunological and physiological properties of this natural substitute for cow's milk. The paper offers a detailed comparative analysis among the protein fractions of DM, CM and human milk, following an extensive proteomic study of the casein and whey proteins of DM performed by narrow pH range 2-DE. The detailed protein composition and structural features reported in this study provide insight into the molecular reasons for the hypoallergenicity of DM. Whole DM might constitute a valid substitute of CM in feeding children with CM protein allergy and it might also constitute the basis for formulas suitable for allergic subjects in the first year of life.

Donkey's milk detailed lipid composition.

Donkey's milk (DM) has recently aroused scientific interest, above all among paediatric allergologists. A deeper knowledge of both proteins and fats in donkey's milk is necessary to evaluate the immunological, physiological and nutritional properties. By using the most refined techniques for fatty acids analysis, the paper offers a detailed comparative analysis of the lipid fractions of DM as well as of human and cow milk, also indicating the distribution of fatty-acid moieties among sn-1/3 and sn-2 positions of the glycerol backbone. In DM the position of fatty acids on glycerol backbone, above all of long chain saturated fatty acids, is very similar to that of human milk: this fact, in conjunction with the relatively high contents of medium-chain triglycerides, makes the lipids in DM, through quantitatively reduced, highly bioavailable. The high PUFA n-3 content of donkey's milk, and especially its low n-6/n-3 ratio, acquires particular interest in subjects affected by cow's milk protein allergy. Whole DM might also constitute the basis for formulas suitable for subjects in the first year of life.

Benefits of donor human milk for preterm infants: current evidence.

It's undoubted that optimum nutrition for term infants is breastfeeding, exclusive for the first six months, then followed by a complementary diet and carried on, if possible, for the first year of life or even more. During the last decades several data confirmed the great advantages of fresh mother's milk use also for feeding very low and extremely low birthweight preterm infants. When mother's milk is unavailable or in short supply, pasteurized donor breast milk is widely used in neonatal intensive care units. Pasteurization partially affects nutritional and immunological properties of breast milk, however it is known that pasteurized milk maintains some biological properties and clinical benefits. The substantial benefits of mother's own milk feeding of preterm infants are supported by strong evidence. However, there is increasing evidence also on specific benefits of donor breast milk. Future research is needed to compare formula vs. nutrient fortified donor breast milk, to compare formula and DM as supplements to maternal milk rather than as sole diet and to compare effects of different methods of heat treatments on donor human milk quality.

Necrotizing enterocolitis: risk factor analysis and role of gastric residuals in very low birth weight infants.

OBJECTIVE: Necrotizing enterocolitis (NEC) usually occurs in very low birth weight infants and is the most common gastrointestinal emergency in the neonatal intensive care unit. Inasmuch as NEC mortality and morbidity are extremely high, early diagnosis becomes essential. Increased gastric residuals are used to define NEC stage, but studies on qualitative and quantitative residual features as markers of NEC risk are still lacking. The primary goal of this analysis was evaluation of the role of gastric residuals in early identification of patients at risk for NEC. The secondary goal was investigation of NEC risk factors, besides prematurity and birth weight. METHODS: In a case-control study, NEC patients were matched with control infants by gestational age and birth weight. Feeding tolerance was assessed by maximum gastric residual volume, maximum residual as percentage of previous feeding, and residual appearance. Mortality and NEC risk factors were also evaluated. RESULTS: In all, 844 very low birth weight infants were admitted to the neonatal intensive care unit during the study period, with an overall mortality before discharge of 14.6%. NEC frequency was 2%. Patent ductus arteriosus was significantly associated with NEC. Mean maximum residual from birth to NEC onset and maximum residual as percentage of the corresponding feed volume were significantly higher in patients than in control infants, as was the percentage of infants with hemorrhagic residuals. CONCLUSIONS: Gastric residuals are a marker of feeding intolerance, and bloody residuals seem to be the best predictor for NEC. For early detection of very low birth weight infants at risk for NEC, both gastric residual volumes and bloody residuals represent an early relevant marker.

Weight growth velocity of very low birth weight infants: role of gender, gestational age and major morbidities.

It's well known that VLBWI fail to thrive, however it's still unclear how gender, GA and morbidities affect growth pattern: aim of this study is to assess the influence of these factors on weight growth. 262 VLBWI were selected. Weight was recorded daily up to 28 days, weekly up to discharge and during 7 scheduled follow-up visits up to 2 years of corrected age. Individual profiles were fitted with a mathematical function suitable to model selected growth milestones and mean distance and velocity curves were drawn. Effects of gender, GA, major-morbidities, nutritional and respiratory support on individual weight growth milestones were estimated using a multivariate linear model. Each of these variables acts differently on weight growth pattern mainly modifying velocity curves characteristics. In particular, infants with major morbidities weight growth impairment-seen on distance curves at 2 years of corrected age-depends on poor weight velocity during a critical period ending within 4th month of postnatal age, for SGA or BPD infants, starting from 5th month of postnatal for severely neurologically impaired infants. These critical periods could be the most appropriate to identify risk factors for weight growth impairment in VLBWI.

Galacto-oligosaccharides are bifidogenic and safe at weaning: a double-blind randomized multicenter study.

OBJECTIVES: The primary objective of this study was to determine the bifidogenic effect of galacto-oligosaccharides (GOS) in a follow-on formula and the effects on other intestinal bacteria. Secondary objectives were the effects on stool characteristics, growth, and general well-being. PARTICIPANTS AND METHODS: In a multicenter, double-blind study, 159 healthy infants, formula-fed at enrollment (at 4-6 months), were randomized to an experimental follow-on formula supplemented with 5 g/L (GOS) (77 infants), or to a standard follow-on formula (control, 82 infants). Infants were evaluated at enrollment (study day 1 = sd1), after 6 weeks (study day 2 = sd2), and after an additional 12 weeks (study day 3 = sd3). At each study day, a fresh stool sample for the bacterial counts was collected, and the growth parameters were measured. At sd2, urinary specimens were collected for the evaluation of urinary osmolarity. RESULTS: At sd2 and sd3, the GOS group had a higher median number (colony-forming units per gram of stool) of bifidobacteria than did the control group (sd2 GOS 9.2 x 10(9) vs control 4.4 x 10(9), P = 0.012); (sd3 GOS 7.2 x 10(9) vs control 2.4 x 10(9), P = 0.027). Other bacteria did not show any significant differences between the 2 groups at all study days. The GOS produced softer stools but had no effect on stool frequency. The urinary osmolarity (mOsm/L) at sd2 was comparable in both groups. Supplementation had no influence on the incidence of gastrointestinal side effects or on the growth of the infants. CONCLUSIONS: These data indicate that the addition of GOS (5 g/L) to a follow-on formula positively influences the bifidobacteria flora and the stool consistency in infants during the supplementation period at weaning. No local or systemic side effects were recorded.

Evaluation of postnatal weight growth in very low birth weight infants.

There is general agreement about the need for longitudinal studies of very low birth weight infants (VLBWI) to evaluate their postnatal growth and to generate distance and velocity charts that allow neonatologists and pediatricians to detect earlier whether a child is not growing adequately. There are no satisfactory growth charts for VLBWI. We analyzed the weight growth of 262 VLBWIs from birth to 2 years of corrected age. Individual growth profiles were fitted with a 7-constant exponential-logistic function suitable for modelling weight growth pattern. After a postnatal weight loss, all VLBWIs showed a late neonatal peak of velocity between the seventh and 21st weeks; the large majority of them also experienced an early neonatal peak between the second and the sixth weeks. Small-for-gestational-age VLBWIs with major morbidities grew less than reference appropriate-for-gestational-age VLBWIs without major morbidities: at 2 years, the difference in weight was about 860 g. The more severe growth impairment in VLBWIs with major morbidities was almost entirely due to the reduced height of the late neonatal peak of velocity. The mathematical function used in this study is expected to be a useful tool to trace model-based longitudinal distance and velocity charts specific for VLBWIs. Moreover, this function also could be used to evaluate to what extent different pathological conditions or nutritional and medical care protocols affect growth kinetics.

Efficacy of donkey's milk in treating highly problematic cow's milk allergic children: an in vivo and in vitro study.

Successful therapy in cow's milk protein allergy rests on completely eliminating cow's milk proteins from the child's diet: it is thus necessary to provide a replacement food. This prospective study investigated tolerance of donkey's milk in a population of 46 selected children with cow's milk protein allergy, for whom it was not possible to use any cow's milk substitute. Thirty-eight children (82.6%) liked and tolerated donkey's milk at the challenge and for the entire duration of follow-up. Catch-up growth was observed in all subjects with growth deficit during cow's milk proteins challenge. The degree of cross-reactivity of immunoglobulin E (IgE) with donkey's milk proteins was very weak and aspecific. Donkey's milk was found to be a valid alternative to both IgE-mediated and non-IgE-mediated cow's milk proteins allergy, including in terms of palatability and weight-height gain.

Magnesium intake, glucose and insulin serum levels in pre-school very-low-birth weight pre-term children.

AIMS: To evaluate cross-sectional associations between dietary magnesium intake and the metabolic pattern of very-low-birth-weight (VLBW, <1500 g) pre-term children, in pre-school years (>2 and <6 years). METHODS AND RESULTS: Fifty-eight Italian children without major congenital malformations/conditions were enrolled; dietary intakes, clinical and (in 34 cases) laboratory characteristics were evaluated. Subjects with lower magnesium intake showed significantly higher fasting glucose, insulin and Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) levels. At simple regression analysis, fasting glucose was significantly associated with magnesium intake (inversely) and catch-up growth (CUG). Fasting insulin and HOMA-IR values were inversely associated with intakes of magnesium and fibres, and directly with Body Mass Index (BMI) and CUG. In a multiple regression model, after adjusting for multiple confounders and fibre intake, magnesium intake was inversely associated with glucose (beta=-0.018; 95%CI -0.026 to -0.010), but not with insulin or HOMA-IR levels. In the same model, dietary fibres remained inversely associated with insulin (beta=-0.075; -0.14 to -0.008) and HOMA-IR levels (beta=-0.06; -0.11 to -0.01). CONCLUSION: These results suggest a significant association between reduced magnesium intake and fasting glucose, and between reduced fibre intake and insulin resistance and this is present even in earlier childhood, and independently of BMI and growth characteristics.

Short-term maternal and neonatal outcomes by mode of delivery. A case-controlled study.

OBJECTIVE: Side-by-side comparisons of short-term maternal and neonatal outcomes for spontaneous vaginal delivery, instrumental vaginal delivery, planned caesarean section and caesarean section during labor in patients matched for clinical condition, age, and week of gestation are lacking. This case-controlled study was undertaken to evaluate short-term maternal and neonatal complications in a healthy population at term by mode of delivery. STUDY DESIGN: Four groups of healthy women, with antenatally normal singleton pregnancies at term, who underwent instrumental vaginal delivery (no. 201), spontaneous delivery (no. 402), planned caesarean section without labor (no. 402) and caesarean section in labor (no. 402) have been retrospectively selected. Outcome measures were maternal and neonatal short-term complications. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. RESULTS: Maternal complications were mostly associated with forceps-assisted and vacuum-assisted instrumental deliveries (OR: 6.9; 95% CI: 2.9-16.4 and OR 3.0; 95% CI 1.1-8.8, respectively, versus spontaneous deliveries). No significant differences in overall complications were observed between spontaneous vaginal deliveries and caesarean sections, whether planned or in labor. By comparison with caesarean sections in labor, instrumental deliveries significantly increased the risk of complications (OR: 3.2; 95% CI: 1.6-6.5). Neonatal complications were also mostly correlated with forceps-assisted and vacuum-assisted instrumental deliveries (OR: 3.5; 95% CI: 1.9-6.7 and OR 3.8; 95% CI 2.0-7.4, respectively, versus spontaneous deliveries). By comparison with caesarean sections in labor, instrumental vaginal deliveries significantly increased the risk of complications (OR: 4.2; 95% CI: 2.4-7.4). CONCLUSIONS: In healthy women with antenatally normal singleton pregnancies at term, instrumental deliveries are associated with the highest rate of short-term maternal and neonatal complications.

Relationship between maternal- and fetal-specific IgE.

AIM: A positive correlation between maternal and cord-blood IgE levels is well documented for total IgEs, but not for specific IgEs. The difficulty in detecting specific cord-blood IgEs is due to their low concentrations, which hinder their dosage by low-sensitivity methods. The study aimed to correlate maternal and foetal specific IgEs against individual cow's milk proteins, detected by highly sensitive and specific techniques. METHODS: Cow's milk specific IgE detection was performed by chemiluminescence on 52 specimens of maternal and cord blood after cow's milk protein separation by 1D and 2D gel electrophoresis. Cow's milk protein (CMP) antigens were identified by mass spectrometry techniques. RESULTS: Specific IgEs for CMPs were found in 25/52 (48.1%) of maternal sera and in 19/52 (37%) of cord-blood sera. In order of decreasing frequency, the proteins found were BSA, IgG heavy chain, caseins and, in a single case, b-lactoglobulin. Positive cord-blood sera in all cases corresponded to a positive maternal result, and maternal and foetal immunoreactivity patterns were closely correlated. Moreover, in no case was there a positive cord-blood response with a negative maternal response. CONCLUSION: The study demonstrates a close relationship between maternal and cord-blood specific IgE patterns. The phenomenon observed could provide a model to elucidate the general production method of foetal IgEs, which might only be produced in the presence of both the corresponding maternal IgE and the related allergen.

The impact of national and international guidelines on newborn care in the nurseries of Piedmont and Aosta Valley, Italy.

BACKGROUND: Care procedures for preventing neonatal diseases are carried out according to nurseries' traditions and may be not consistent with the evidence based medicine issues. METHODS: A multi-centric survey was conducted in 2 Regions located in NW Italy (Piedmont and Aosta Valley) in order to collect information on some healthy newborn care procedures. During 2001, a questionnaire was sent to the chief pediatrician in charge to the all 33 nurseries of the region asking the methods used during 2000 as prevention of ophthalmia neonatorum, early and late hemorrhagic disease of newborn, umbilical cord care and recommendations of vitamin D administration. Thereafter, during 2004 the same questionnaire was sent to the 34 chief pediatrician of nurseries to evaluate if the procedures were changed during 2003 according to guidelines. The nurseries care for 32,516 newborns in 2000 and 37,414 in 2003. RESULTS: Aminoglycoside eyes drops as prevention of ophthalmia neonatorum were the first choice in both periods (23 out 33 nurseries in 2000 and 24 out 34 in 2003 p > 0.05; the corresponding figures for newborns were 18,984 out 32,516 newborns vs. 28,180 out of 37,414 p < 0.05). The umbilical cord care was carried out with alcohol in 12/33 centers (13,248 newborns) and dry gauze in 3/33 centers (2,130 newborns) in 2000, the corresponding figures in 2003 were 6/34 centers (p > 0.05), (6,380 newborns, p < 0.05) and 12/34 centers (p < 0.05), (18,123 newborns, p < 0.05). The percentage of newborns receiving of i.m. vitamin K. at birth increased during the study period (15,923/32,104 in 2000 vs. 19,684/37,414 in 2003, p < 0.01), but not the number of nurseries (16 in 2000 and 17 in 2003 p > 0.05). The numbers of parents of newborns who receive the recommendations of oral vitamin K during the first months life decreased from 2000 (25,516/30,606) to 2003 (29,808/37,414, p < 0.01) as well as for Vitamin D recommendation (14,582/30,616 in 2000 vs. 11,051/37,414 in 2003, p < 0.01). Oral vitamin K during the first months of life was recommended by 25 nurseries in 2000 and 27 in 2003 (p > 0.05), the corresponding figures for Vitamin D were 15 and 14 (p > 0.05). CONCLUSION: In the present study a large variability of procedures among the nurseries was observed. During the study periods, guidelines and evidence based medicine issues have only partially modified the neonatal care procedures In Piedmont and Aosta Valley nurseries. These observations suggest to implement local forum/consensus conference to standardized procedures as much as possible.

Differences in size at birth are determined by differences in growth velocity during early prenatal life.

Physiologic interindividual differences in neonatal size are traditionally thought of as determined by differences in fetal growth occurring only in the second half of pregnancy. Whether possible differences in early intrauterine growth velocity are the effect of random growth fluctuations or may affect size at birth is still debated. This article aims at evaluating to what extent differences in neonatal size are accounted for by differences in fetal growth velocity. We analyzed the fetal growth of 130 healthy singletons for whom head (HC) and abdomen (AC) circumferences and femur diaphysis length (FDL) longitudinal profiles were available, together with the measures of weight (BW), length (BL), and head circumference (BHC) at birth. Individual profiles were fitted with ad-hoc models. Neonatal traits were transformed into standard deviation scores (SDS). Neonates in the upper third of BW-SDS distribution (3618+/-43 g, mean+/-SEM) had, at 22 wk of gestational age, AC growth velocity higher by 0.55+/-0.10 mm/wk than those in the lower third (2902+/-36 g). Neonates in the upper third of BL-SDS distribution (51.7+/-0.21 cm) had, at 20 wk, FDL growth velocity higher by 0.11+/-0.05 mm/wk than those in the lower third (48.2+/-0.18 cm). Neonates in the upper third of BHC-SDS distribution (35.7+/-0.13 cm) had, at 18 wk, HC growth velocity higher by 0.57+/-0.20 mm/wk than those in the lower third (33.3+/-0.11 cm). The differences in growth velocity remain constant throughout the second and third trimester for AC, and tend to vanish in the third trimester for HC and FDL. The differences in fetal growth velocity, which in our study were observed as early as mo 4, suggest that the genetic component plays an important role in fetal growth and is precociously expressed.