Loss of triglycerides and carotenoids in human milk after processing.

OBJECTIVE: Human milk (HM) is considered to be the best nutrition for preterm infants. However, storage, heating or tube feeding can cause a decline in essential nutrients, which can lead to the loss of antioxidant vitamins, resulting in an increased risk for oxygen radical diseases. Recently we found that carotenoids, present in human milk, can play a role in the antioxidant protection of preterm infants. In this study we evaluated the effect of processing HM and infant formula on the triglycerides and carotenoid concentrations. DESIGN: The triglyceride, alpha- and beta-carotene, lutein and lycopene concentrations of 30 samples of mature HM of mothers who delivered a term infant and 10 samples of infant formula were measured after refrigeration, freezing, microwave heating and tube feeding with and without exposure to normal light and phototherapy, imitating the clinical feeding routine in the NICU. RESULTS: After tube feeding triglyceride, lutein and beta-carotene concentrations decreased with 33%, 35% and 26% respectively. The decrease in triglycerides in HM accounts for 16% of the total caloric intake of neonates. Triglyceride and carotenoid concentrations in HM remained stable after refrigeration, freezing or low temperature microwave heating, except for lutein which decreased after refrigeration and freezing. In infant formula no differences were found. CONCLUSIONS: Mature human milk can be stored safely in a freezer and heated in a microwave oven without loss of fat or carotenoids. The clinically important loss of fat during tube feeding is probably the most important contributing factor to the decrease in lutein and beta-carotene in tube feeding, with only a small role for peroxidation during light-exposure.

Body temperature measurement in VLBW infants by continuous skin measurement is a good or even better alternative than continuous rectal measurement.

BACKGROUND: An inadequate body temperature in preterm infants influences morbidity and mortality. Continuous rectal measurement is a reliable method to measure body temperature but might have adverse effects and is even contra-indicated in case of low platelets or necrotising enterocolitis. A save and non-invasive method to measure body temperature is the transcutaneous 'zero heat flow' method. AIM: We hypothesised that for monitoring body temperature in very low birth weight (VLBW) infants, central measurement of temperature by way of the zero heat flow principle is just as reliable as rectal temperature. METHODS: Twenty-six infants, birth weight between 520 g and 1250 g, gestational age 25.28-32.28 weeks were provided with an insulated continuous skin probe with 'zero heat flow' and a continuous rectal probe. Both measurements were registered every hour over a period of 48 h. The sample size was calculated to detect a difference of less than or equal to 0.20 degrees C. RESULTS: 1205 of the 1248 temperature measurements were analysed. At any moment, skin temperature was higher or equal when compared to rectal temperature. Mean skin temperature was 0.13 degrees C (SD 0.33) higher than mean rectal temperature (t-test, p < 0.001). Correlation between rectal and skin temperature was 0.82 (p

Postnatal changes in plasma ceruloplasmin and transferrin antioxidant activities in preterm babies.

Postnatal changes in plasma ceruloplasmin ferroxidase and transferrin iron-binding antioxidant activity were studied in 10 healthy preterm babies during the first 6 weeks of life. Ceruloplasmin levels and ceruloplasmin ferroxidase activity were low at birth, remained stable for the first 3 weeks, and increased between 3 and 6 weeks. The transferrin levels were also low at birth, and this finding persisted throughout the 6-week study period. However, although the plasma iron-binding antioxidant activity was correspondingly low at birth, it thereafter rose and remained high. In four cord blood samples, but not in subsequent postnatal samples, peroxidation was actually stimulated in the assay measuring plasma iron-binding antioxidant activity. We have previously shown that this phenomenon is probably due to the presence of non-protein-bound iron.

Plasma proteins in acute and chronic lung disease of the newborn.

This study compared plasma levels of albumin, transferrin, and ceruloplasmin in well preterm babies (n = 21) with those with respiratory distress syndrome (RDS, n = 13) and chronic lung disease (CLD, n = 13) over the first 28 postnatal days. Plasma lipid peroxidation, total radical trapping capacity (TRAP assay), and iron binding antioxidant capacity were also measured. In RDS and CLD albumin levels were decreased on days 1, 4 and 10; on day 10 albumin was lower in CLD compared to RDS (p < .05). After day 10 the levels were similar in all groups. The transferrin levels showed a similar trend. Ceruloplasmin levels did not differ, except for a higher day 28 level in CLD (p < .05). Albumin levels significantly decreased with increasing FiO2 and duration of oxygen therapy (within patient r = -0.30, p < .05 and r = -0.51, p < .005, respectively). On day 10, increasing oxygen therapy increased plasma lipid peroxidation (r = +0.49, p < .01), which was also significantly related to lower plasma protein levels (r = -0.42, p < .01). Lower plasma albumin and transferrin lowered the TRAP and iron binding antioxidant capacity, respectively (r = +0.36, p < .05, and r = +0.41, p < .005). Prediction of CLD using day 10 albumin levels had a specificity of 94%, but a sensitivity of only 50%. The interaction between oxygen toxicity and high ventilation pressures in immature babies appears to lower plasma proteins by increasing pulmonary permeability. The lower plasma albumin level was not useful in predicting the development of CLD; however, the fall in plasma transferrin and albumin will further decrease the preventive and chain-breaking antioxidant capacity of plasma of these ill babies.

Vitamin E status in preterm infants: assessment by plasma and erythrocyte vitamin E-lipid ratios and hemolysis tests.

BACKGROUND: Vitamin E is an essential component of the antioxidant defenses, but supplementation can have side effects in the preterm infant. Careful monitoring of vitamin E status is thus essential, however no consensus has been reached on the best clinical method. METHODS: In 47 healthy preterm infants, several methods for assessment of vitamin E status were evaluated: plasma and erythrocyte vitamin E levels were measured, vitamin E lipid ratios were calculated, and two variations of the hydrogen peroxide hemolysis test were conducted. RESULTS: At birth, the plasma and erythrocyte vitamin E levels were low. After birth, the plasma levels rose gradually, whereas the erythrocyte levels remained low. In contrast, the vitamin E-total-lipid ratio was in the low normal range from birth onwards. Vitamin E-lipid ratios using two lipid components (cholesterol with triglycerides, or cholesterol with phospholipids) or one lipid component (cholesterol) correlated with the vitamin E-total-lipid ratio with a good sensitivity and specificity. The susceptibility of erythrocytes to hydrogen peroxide-induced damage (measured as potassium release or malondialdehyde production) was high at birth and declined after birth. However, this susceptibility did not correlate with plasma or erythrocyte vitamin E levels or vitamin E-total-lipid ratio, and the hydrogen peroxide hemolysis test is not a reliable indicator of vitamin E status in preterm infants. CONCLUSIONS: Our study indicated that a gold standard for clinical assessment of vitamin E status in preterm infants is yet to be found.

Serratia marcescens infections in neonatal departments: description of an outbreak and review of the literature.

An outbreak of colonization and infection with Serratia marcescens occurred in a neonatal intensive care unit (NICU). S. marcescens was isolated from five preterm infants (gestational age 25-30 weeks). Two infants developed septicaemia, which were both fatal, and one infant (the presumed index case) had conjunctivitis due to S. marcescens. Two infants were colonized without clinical signs of infection. All infants were treated with antibiotic regimens including ciprofloxacin and gentamicin. The DNA fingerprints of isolates were determined by enterobacterial repetitive intergenic consensus primers by the polymerase chain reaction. This showed that a single strain had spread in the NICU. An extensive investigation pointed to an infant born from a mother with an intra-uterine infection after prolonged rupture of foetal membranes as a presumed source of the outbreak. A reservoir, other than the infected or colonized infants and their immediate vicinity, was not found, with the sole exception of the waste jar of a Na+/ K(+)-analysis apparatus. Containment of the outbreak was achieved by closure of the NICU for new admissions, strict hygienic measures and cohort nursing of the infected and colonized infants. It was considered especially important to handle the infants with gloves, since frequent hand carriage of staff with S. marcescens was found when gloves were not used.

Uric acid and ascorbic acid redox ratios in plasma and tracheal aspirate of preterm babies with acute and chronic lung disease.

This study compared plasma redox ratios of uric acid and ascorbic acid in well preterm babies with those with respiratory distress syndrome (RDS) and chronic lung disease (CLD), and investigated the relationship between these ratios and their respective measurements in tracheal aspirate. On day 1 after birth, plasma allantoin and allantoin/uric acid ratio were elevated in CLD (p < .05), and both markers of oxidative stress enabled early prediction of development of CLD (sensitivity and specificity: 54 and 83%, respectively). The relation between allantoin production and oxidative stress is supported by the correlation between the allantoin level and oxygen therapy in both RDS and CLD (p < .05). Reduced and oxidize ascorbic acid in plasma decreased postnatally in all groups and their redox ratio remained stable. Uric acid and ascorbic acid redox ratios were significantly elevated in tracheal aspirates compared to plasma samples (p < .05), and there was a strong positive correlation between both ratios (p < .005). These markers may be useful in monitoring babies with respiratory distress.

Markers of oxidative stress and antioxidant activity in plasma and erythrocytes in neonatal respiratory distress syndrome.

Markers of oxidative stress and antioxidant activity in plasma and erythrocytes were studied for 14 d after birth in infants with neonatal respiratory distress syndrome (n = 9) and controls (n = 36). In plasma, the total radical trapping antioxidant capacity and the chain-breaking antioxidants vitamin C, sulfhydryl groups and bilirubin were similar. The differences in uric acid levels were not consistent, but vitamin E levels and vitamin E/total-lipid ratio were lower in the neonatal respiratory distress group (p < 0.01). In erythrocytes, the antioxidant enzymes glutathione peroxidase, glutathione reductase and superoxide dismutase did not differ postnatally. Indicators of oxidative damage in plasma (sulfhydryl/protein ratio and thiobarbituric acid reactive substances) showed the same postnatal course in both groups and were not influenced by oxygen therapy. In erythrocytes the reduced/oxidized glutathione ratio showed no consistent differences. In conclusion, this study, using erythrocytes and plasma, does not provide convincing evidence of oxidative damage and diminished antioxidant defenses in preterm infants with neonatal respiratory distress syndrome.

Red blood cell glutathione and plasma sulfhydryls in chronic lung disease of the newborn.

We compared the postnatal changes (days 1-28) in red blood cell glutathione and plasma sulfhydryl content in preterm babies developing chronic lung disease (CLD, n = 13) to those in babies with respiratory distress syndrome (RDS, n = 13) and control babies (n = 21). There were no initial differences in these measurements between the three groups. However, on day 28 in CLD and RDS the red blood cell glutathione was decreased compared to the control babies (p < 0.05). In CLD, there was a significant correlation between reduced/oxidized glutathione and (i) maximal FiO2 (r = -0.69, p < 0.05) and (ii) minimal a/A ratio (r = +0.73, p < 0.005). On day 28, although the plasma sulfhydryl level did not differ between the groups, the sulfhydryl/total protein ratio was decreased in CLD (p < 0.05). The late decrease in both glutathione and sulfhydryl/total protein ratio in babies with CLD suggests that oxidative stress is still ongoing at 28 days after birth and that the antioxidant capacity of their blood is still diminished at this time.

Influence of long chain unsaturated fatty acids in formula feeds on lipid peroxidation and antioxidants in preterm infants.

The influence of long chain polyunsaturated fatty acids (LCP) in formula feeds on lipid peroxidation and antioxidants was studied in 35 healthy preterm infants (gestational age 30-35 wk) during the first 6 postnatal weeks. Infants received a preterm formula supplemented with n-3 LCP (LCP group, n = 13), or standard preterm formula (NO-LCP group, n = 15); 7 infants fed human milk served as a reference group. With LCP supplementation, erythrocyte C22:6n-3 levels were stable; without supplementation, the levels declined (difference p < 0.001). LCP supplementation did not decrease vitamin E or C levels, or increase lipid peroxidation products (thiobarbituric acid-reactive substances) in plasma. In erythrocytes, LCP supplementation did not markedly influence the reduced/oxidized glutathione ratio; however, the susceptibility to H2O2-induced oxidative stress was reduced. Our results suggest that healthy preterm infants are able to cope with any extra peroxidative stress produced by n-3 LCP supplementation. However, these findings might not be generally applicable to other formulas containing LCP supplements.

Nonprotein-bound iron in postasphyxial reperfusion injury of the newborn.

OBJECTIVE: To investigate if the availability of nonprotein-bound iron after birth asphyxia is related to the severity of the postasphyxial injury and neurodevelopmental outcome. METHODS: Nonprotein-bound iron (bleomycin assay) and thiobarbituric-acid-reactive species, an index of oxidative lipid damage, were measured in plasma of 50 newborn infants (gestational age > 34 weeks) between 0 to 8 hours, 8 to 16 hours, and 16 to 24 hours after birth. Three groups were compared: healthy infants (n = 20), moderately asphyxiated infants (n = 15), who were neurologically normal during the first 24 hours after birth and severely asphyxiated infants (n = 15), who developed abnormal neurological signs in the first 24 hours after birth. RESULTS: In the severely asphyxiated infants, liver enzymes, creatinine, urea, and uric acid concentrations were significantly elevated. Eleven severely asphyxiated infants were brain-damaged, 9 of them died during the neonatal period. Nonprotein-bound iron was detectable in 30% of the control, 60% of the moderately asphyxiated, and 80% of the severely asphyxiated infants. During the whole study period nonprotein-bound iron concentration was significantly elevated in severely asphyxiated infants as compared with controls. Three of the four severely asphyxiated infants who had a normal outcome at 1 year of age, had no detectable nonprotein-bound iron during the study period. Stepwise logistic regression analysis with neurodevelopmental outcome at 1 year of age (normal versus adverse/death) as dependent variable and all the measured parameters for organ damage as independent variables revealed that the nonprotein-bound iron concentration at 0 to 8 hours after birth was the most significant variable and at the same time the only variable that entered the model, in relation to neurodevelopmental outcome. Thiobarbituric-acid-reactive species tended to be higher in severely asphyxiated infants, suggesting oxidative lipid damage. CONCLUSION: Nonprotein-bound iron may play an important role in oxidative damage-mediated postasphyxial brain injury and subsequent neurodevelopmental outcome.

Influence of plasma preparations and donor red blood cells on the antioxidant capacity of blood from newborn babies: an in vitro study.

We investigated in an in vitro transfusion model the early effects of plasma preparations and donor red blood cells on the antioxidant capacity of the cord blood from babies. Addition of pasteurized plasma protein solution to plasma from babies decreased the peroxyl radical trapping capacity (p < 0.02). In contrast, fresh frozen plasma did not lower this capacity. Addition of adult donor red blood cells to the babies' red blood cells did not significantly decrease the glutathione-recycling capacity of the blood. On the basis of these in vitro results we hypothesize that the use of resuscitation fluids with low antioxidant capacity may temporarily decrease the ability of the baby to catabolize reactive oxygen species.

Postnatal changes in plasma chain-breaking antioxidants in healthy preterm infants fed formula and/or human milk.

Concentrations of chain-breaking antioxidants were studied in the first 6 postnatal weeks in 29 healthy preterm infants (gestational age 30-35 wk). Vitamin C, uric acid, and sulfhydryl groups declined, whereas vitamin E rose and bilirubin followed its typical biphasic postnatal course. The influence of these changes on the plasma peroxyl radical trapping capacity was assessed in vitro (TRAP assay). The trapping capacity decreased postnatally and this appeared to be related to the coincident fall in uric acid concentrations. Results did not differ between babies fed with only preterm formula (n = 12) and those fed predominantly with human milk (n = 6), except for higher bilirubin and TRAP values in the breast-fed infants. There are major postnatal changes in the concentrations of the plasma chain-breaking antioxidants and this may influence the susceptibility of the preterm baby to oxygen toxicity.

Lipid peroxidation in human milk and infant formula: effect of storage, tube feeding and exposure to phototherapy.

Preformed lipid peroxidation products present in the feed may contribute to the total reactive oxygen radical load infants have to deal with and may play a role in the pathogenesis of necrotizing enterocolitis and bronchopulmonary dysplasia. In this study, the occurrence of lipid peroxidation in human milk and feeding formulas for preterm babies was evaluated in vitro. Free linoleic acid (18:2) and its hydroperoxide (18:2OOH) were measured by gas chromatography-mass spectrometry and the concentration of 18:2OOH and the 18:2OOH/18:2 ratio were used as indices of peroxidation. In all feeds peroxidation products were present, but the proportion of peroxidized 18:2 was greater in infant formula. Storage of human milk (+4 degrees C for four days) increased lipid peroxidation. Exposure to light during tube feeding increased peroxidation in infant formula but not in human milk. Different procedures for preparation, storage and feeding may decrease the concentration of these potentially toxic peroxidized lipids in human milk and infant formula.

Effect of an exchange transfusion on plasma antioxidants in the newborn.

The effect of an exchange transfusion on antioxidants in the plasma of newborns with rhesus hemolytic disease was studied. The antioxidant concentrations in donor blood were similar to normal adult values except for the lower vitamin C concentrations. Exchange transfusion decreased the newborns' iron and ferritin levels and increased their ceruloplasmin and transferrin (primary antioxidants) concentrations and latent iron-binding capacity. The changes in secondary antioxidant concentrations were variable; uric acid and thiols were stable, vitamin C and bilirubin fell, and vitamin E rose. The total peroxyl-radical trapping capacity of the secondary antioxidants did not change significantly. The fall in levels of thiobarbituric acid reactive substances, an index of lipid peroxidation, was related to the lower levels present in the donor blood. Exchange transfusion rapidly produced variable changes in the concentrations of prooxidant and antioxidant substances in plasma and may thus influence free radical metabolism in the newborn.

Iron overload, free radical damage, and rhesus haemolytic disease.

To test the hypothesis that iron overload induces free radical damage in rhesus haemolytic disease (RHD), cord blood plasma of babies with RHD was compared with that of controls matched for gestational age. Babies with RHD had higher ferritin levels, lower latent iron-binding capacity, increased concentrations of lipid-peroxidation products, and low vitamin C levels. Plasma of 3 RHD babies did not inhibit peroxidation stress. These findings, of iron overload and free radical damage, have implications for treatment of RHD.

The total free radical trapping ability of cord blood plasma in preterm and term babies.

The interaction between various antioxidants may be important in protecting the newborn baby against oxygen toxicity. We studied the total radical trapping capacity of the antioxidants in plasma (TRAP) and compared the TRAP level in the preterm and term baby (cord blood) with that in adults. In addition, the concentrations of various known antioxidants were measured and the theoretical contribution of these antioxidants to the TRAP calculated. The measured and calculated TRAP were higher in the newborn babies than the adults. The uric acid concentration was similar in the three groups but the vitamin C concentration was higher and the vitamin E and sulfhydryl concentrations were lower in the newborn babies. In contrast to the adult group, the measured TRAP in the newborn babies did not correlate with the calculated TRAP. This may be due to differences in inhibition or recycling of antioxidants in the newborn and adult groups. Theoretical considerations showed that there may be a large unidentified group of antioxidants that contribute to measured TRAP in plasma. Bilirubin and beta-carotene were measured (higher and lower concentrations, respectively, in the newborn) in an attempt to identify these antioxidants. The efficient plasma radical trapping capacity in the cord blood may partly compensate for deficiencies in other components of the antioxidant defenses, e.g. cellular enzymes, at the time of birth.

Human milk vitamin content after pasteurisation, storage, or tube feeding.

We investigated the effect of the composition of the storage container, Holder pasteurisation, and conditions during tube feeding on the concentration of selected vitamins in human milk. Though the fat soluble vitamins A, D, and E were not affected, the concentration of several water soluble vitamins decreased. The lower vitamin C concentration of milk stored in polypropylene containers compared with milk stored in glass containers (29%) was not significant. Holder pasteurisation significantly lowered the concentrations of vitamins C (36%), folacin (31%), and B6 (15%). Tube feeding significantly lowered the concentrations of vitamins C (44%) and B6 (19%), and exposure to phototherapy seemed to lower the vitamin C concentration (53%) further. Low birthweight infants have increased vitamin requirements. Vitamin losses in expressed human milk before or during feeding may increase the incidence of vitamin deficiencies in these babies.

[Diagnosis of hereditary metabolic disorders in newborn and young infants]. / Herkenning van erfelijke stofwisselingsstoornissen bij de pasgeborene en de jonge zuigeling.

This report describes anamnestic, physical and biochemical data in the neonatal period which have led to the diagnosis of an inborn error of metabolism in 12 children. Even in hospital laboratories without facilities for elaborate biochemical assays, adequate investigations permitting timely referral of the patient to a more specialised hospital can often be carried out. Most important is the constant awareness of the pediatrician in case of serious illness which cannot be easily explained by more common causes. The similarity of the clinical presentation and often the coexistence of neonatal sepsis and inborn error of metabolism are stressed. Even when treatment is not possible or has not been successful, a correct diagnosis is important for genetic counseling and antenatal diagnostic procedures.